[Cyclin A1 affects the invasion, metastasis, and prognosis of hepatocellular carcinoma].

Objective: To investigate the effect of cyclin A1 on the invasion, metastasis, and prognosis of hepatocellular carcinoma (HCC). Methods: Immunohistochemistry (IHC) was used to detect the expressional condition of cyclin A1 in HCC and paraffin-embedded non-tumor adjacent tissues. Kaplan-Meier method was used for the survival analysis of patients with HCC. Western blot (WB) was used to detect the expression of cyclin A1 in HCCLM3 and QGY-7703 cells. Scratch wound healing assay, transwell migration, and invasion assay were used to detect the effect of cyclin A1 overexpression on cell migration and invasion ability. WB was used to detect changes in the expression of matrix metalloproteinase (MMP) 2, MMP9, and vascular endothelial growth factor (VEGF) after overexpression of cyclin A1. Measurement data were compared using a t-test and analysis of variance. Count data was measured using χ (2) test and the Log-rank method was performed for survival analysis. Results: Cyclin A1 expression rates were higher in the tissues of HCC patients with recurrent metastasis than in the tissues of patients without recurrent metastasis (60.42% vs. 46.81%, χ (2) = 4.711, P < 0.05). The overall postoperative survival time (OS) and disease-free survival (DFS) were shorter in patients with high cyclin A1 expression than those with low cyclin A1 expression (45.9 months vs. 53.1 months; 42.9 months vs. 51.3 months, and P < 0.01). The postoperative OS and DFS were shorter in patients with high cyclin A1 expression and recurrent metastasis than those with low cyclin A1 expression without recurrent metastasis (31.7 months vs. 43.9 months; 18.0 months vs. 31.5 months, and P < 0.05). HCCLM3 and QGY-7703 cells were higher in the cyclin A1-pEX group than in the empty vector (vector) group (1.56 ± 0.06 vs. 0.18 ± 0.01, t = 18.75, P < 0.001; 1.31 ± 0.05 vs.0.37 ± 0.02, t = 15.17, P < 0.001). The migrated distances of HCCLM3 cells in the cyclin A1-pEX group and the vector group were (536.7 ± 14.5) µm and (327.3 ± 9.3) µm, t = 11.84, P < 0.05, respectively, while the migrated distances of QGY-7703 cells in the two groups were (916.7 ± 35.3) µm and (320.0 ± 20.8) µm, t = 13.54, P < 0.01. The migrated numbers of HCCLM3 cells in the cyclin A1-pEX group and vector group were (37.3 ± 2.4) and (7.0 ± 1.2), t = 12.67, P < 0.001, and the number of invasive cells was (73.7 ± 4.1) and (12.6 ± 1.5), t = 12.36, P < 0.001, respectively. The migrated numbers of QGY-7703 cells in the two groups were (153.3 ± 6.0) and (17.7 ± 3.7), t = 17.59, P < 0.001, and the number of invasive cells was (45.0 ± 2.9) and (9.3 ± 1.5), t = 10.66, P < 0.001, respectively. The expression levels of MMP2, MMP9, and VEGF in HCCLM3 and QGY-7703 cells were significantly higher in the cyclin A1-pEX group than those in the vector group (P < 0.05). Conclusion: Cyclin A1 plays an important role in HCC invasion and metastasis, but HCC patients with high cyclin A1 expression have a poor prognosis. Hence, cyclin A1 has high guiding significance for evaluating patient prognosis.

[Clinical outcomes of radial artery as the second arterial conduit in coronary artery bypass].

Objective: To examine the clinical experience and outcomes of coronary artery bypass grafting (CABG) using radial artery as the second arterial graft. Methods: Totally 585 patients in whom both left internal thoracic artery and radial artery as arterial conduits were used in CABG in Department of Cardiovascular Surgery, Nanjing Hospital Affiliated to Nanjing Medical University from April 2008 to August 2019 were consecutively enrolled. There were 436 males and 149 females, aging (63±10) years (range: 36 to 86 years). There were 40.7% (238/585) of patients had diabetes and 75.6% (442/585) of them had multivessel disease (two-vessel or three-vessel diseases). From January 2017, transit time flow measurement was performed on every patient. Demographic and perioperative data were retrospectively collected, as well as follow-up data for patients who underwent CABG from January 2014 to August 2019. Analysis were made on their early and late outcomes. Results: 81.9%(479/585) Most patients in this cohort (81.9%) received on-pump CABG and 11 patients had intraoperative intro-aortic balloon counterpulsation (prior to CABG) support. Forty-three patients had concomitant valve procedures. The number of distal anastomosis was 3.6±0.9 (range: 2 to 6) and number of arterial distal anastomosis was 2.1±0.3. Radial artery was anastomosed to left obtuse marginal artery in 95.8% (560/585) patients. All target vessels for radial artery conduit had significant proximal stenosis (>70%) and 72.5% (424/585) of target vessels had proximal stenosis which was >90%. Intraoperative transit-time flow measurement of 151 cases showed that radial artery conduits had a flow of (29.8±10.2) ml/minutes (range: 10 to 150 ml/min), and pulsatility index of 2.5±1.4 (range: 0.7 to 5.0). There was no operative death. Two in-hospital death occurred more than 30 days after index surgery. There was no perioperative myocardial infarction. There were 188 patients who received CABG from January 2014 to August 2019 followed-up for a median duration of 3.2 years. There were two noncardiac death. No patient had myocardial infarction or to receive myocardial revascularization. Conclusions: Radial artery as the second arterial conduit is a safe and effective strategy for CABG. Good selection of target vessel and intraoperative transit-time flow measurement may help achieve good patency, as well as the short and mid-term outcome.

[Significance of expression of Myc-induced nuclear antigen 53 in hepatocellular carcinoma and its relationship with patient prognosis].

Objective: To detect the expression of Myc-induced nuclear antigen 53 (Mina53) and liver tissue >5 cm from the edge of the tumor (LTM5), and analyze its relationship with tumorigenesis, clinicopathological characteristics, and patient survival and prognosis in hepatocellular carcinoma (HCC). Methods: The expression levels of Mina53 mRNA and protein in 18 pairs of fresh HCC and LTM5 were assessed by qRT-PCR and Western blot, respectively. The expression of Mina53 in 284 pairs of HCC and LTM5 sample was determined by immunohistochemistry. Paired-sample t-test was used for the comparison of measurement data among groups, and heterogeneity of variance was tested using Wilcoxon rank-sum test. χ (2) test was used for the comparison of measurement data among groups. Kaplan-Meier method and log-rank test were used for survival analysis. Cox regression model was used for single factor and multi factor analysis. Results: The relative expression levels of Mina53 mRNA and protein in 18 fresh HCC tissues were significantly higher than those in LTM5 tissues (mRNA: -4.41 ± 1.48 and -5.93 ± 1.65, t = 3.100, P = 0.007; protein: 1.12 ± 0.29 and 0.46 ± 0.21, t = 10.616, P < 0.001). The relative expression level of Mina53 in 284 HCC tissues was higher than that of LTM5 (z = -18.739, P < 0.001). The expression level of Mina53 was associated with tumor size (χ (2) = 5.474, P = 0.019), vascular invasion (χ (2) = 8.965, P = 0.003), pathological grade (χ (2) = 12.006, P = 0.002), and TNM stage (χ (2) = 16.686, P < 0.001). The overall postoperative survival time and disease-free survival time of patients with high expression of Mina53 (28.5 months and 22.7 months, respectively) were shorter than those with low expression (33.0 months and 31.8 months, respectively) (P < 0.05) in HCC. Cox multivariate regression analysis showed that Mina53 and multiple tumors were independent prognostic factors affecting the overall postoperative survival time and disease-free survival time of HCC patients (P < 0.05). Conclusion: Mina53 may play an important role in the occurrence of HCC and participate in the process of tumor growth as well as invasion and metastasis. The high expression of Mina53 signifies that the patient has a poor prognosis and thus can be used as a potential marker for judging the prognosis of HCC patients.

[Effect of perioperative multi-day low dose ketamine infusion on prevention of postmastectomy pain syndrome].

Objective: To investigate the effects of multi-day low dose ketamine infusion for postmastectomy pain syndrome (PMPS) after breast cancer surgery. Methods: This study was a prospective randomized controlled trial. From June 2015 to May 2016 in Affiliated Yiwu Hospital of Wenzhou Medical University, 66 patients with breast cancer surgery were randomly divided into control group (group C) and ketamine group (group K). Patients in group K were infused with 0.5 mg/kg of ketamine mixed in 250 ml of 0.9% normal saline in 1 h daily for 7 days. Patients in group C were infused the same dose of 0.9% normal saline. Anesthesia induction in both groups were given intravenous midazolam, sufentanil, propofol, vecuronium and intermittent positive pressure ventilation after tracheal intubation, anesthesia was maintained with propofol and remifentanil. After awakening, all patients were monitored in postanesthesia care unit (PACU) and given patient-controlled intravenous analgesia(PCIA). Pain scores were assessed using visual analogue scales (VAS) during PACU, 4 h, 24 h and 2-5 d after surgery, simultaneously analgesic requirement were recorded. Patients were evaluated Hospital Anxiety and Depression Scale (HADS) 5 d after surgery . The patients were followed up for 6 months. At 3 m, 6 m after surgery, the incidence of PMPS, the level of pain, pain site and HADS scale were assessed. Results: The VAS score uring PACU, 4 h, 24 h and 2-5 d after surgery in group K( (2.5±0.8), (2.4±0.5), (2.4±0.5), (2.0±0.4), (1.5±0.5), (1.0±0.4), 1(1), respectively) was lower than those in group C ((2.9±1.0), (2.9±0.6), (2.6±0.5), (2.3±0.5), (1.8±0.6), (1.5±0.5), 1(0), respectively). There was statistically difference between the two groups (all P<0.05). The consumption of analgesics required at each time postoperation in group K were also lower than that of group C(all P<0.05). Followed up for 6 months, 2 lost in group C, 1 lost in group K. The incidence of PMPS in group K at 3 months and 6 months after surgery was significantly lower(25% and 22%) than that in group C(52% and 45%)(χ(2)=4.729, 3.842, all P<0.05). There were no significant difference in pain level and site between two groups of PMPS patients (all P>0.05). There were no significant difference of HADS scale preoperative and 5 d after surgery between two groups (all P>0.05); and HADS scale in group K at 3 m and 6 m after surgery was significantly lower than that in group C(all P<0.05). Conclusion: Perioperative continuous multi-day low dose ketamine infusion can effectively reduce the incidence of PMPS after breast cancer surgery.

[Efficacy of mitral valve repair versus replacement in severe ischemic mitral regurgitation].

Objective: To compare the mortality, survival rate and the therapeutic efficacy between mitral valve repair and replacement as treatment for severe ischemic mitral regurgitation (IMR), and explore the middle- and long-term outcomes. Methods: Between January 2000 and January 2016, 378 patients with severe IMR underwent coronary artery bypass grafting (CABG) combined with mitral valve repair (n=162) or mitral valve replacement (n=216) in the Department of Cardiovascular Surgery of Nanjing First Hospital. Clinical data, in-hospital morbidity and mortality of patients were retrospectively reviewed. The patients were followed up for the long term survival rate, heart function and re-admission. Results: No statistically significant differences of baseline data and operation details were found between the two groups except for left ventricular end-diastolic diameter[(61.3±10.2)mm in replacement group vs (56.2±9.0)mm in repair group, P<0.001]. Seven patients died during the perioperative period, with a total operation mortality of 1.9%.No significant difference of mortality was found between the two groups (5 cases in the replacement group and 2 cases in the repair group). The early outcome after the surgery showed that the rate of low cardiac output and ventricular arrhythmia of patients were significantly higher in the replacement group compared with the repair group (both P<0.05). The mortality of patients received mitral valve replacement was better than who received mitral valve repair when left ventricular end-diastolic diameter was over 65 mm (5.9% vs 10.0%, P=0.036). No significant differences were observed between the two groups in the middle- and -long term survival rate (87% for replacement group vs 85% for repair group, P=0.568). The follow-up time was 1-85 (52.8±21.5) months and the follow-up rate was 93%. The rate of valve-related complications was significantly higher in the repair group compared with the replacement group (8.82% vs 3.82%, P=0.003). Conclusions: We should choose the surgical methods carefully (replacement or repair) for severe IMR patients according to degree of left ventricular remodeling and pathological changes of mitral valve. Mitral valve replacement with preservation of the subvalvular apparatus is a safe and effective surgical alternative for mitral valve repair, especially for patients with complications or complex reflux.

[Expression of ATAD2 in different liver lesions and its clinical significance].

Objective: To examine the expression of ATAD2 in different liver lesions and its clinical significance. Methods: ATAD2 expression in 60 hepatocellular carcinoma (HCC) surgical specimens (49 of which have concurrent liver cirrhosis), 43 HCC biopsy specimens, 2 high-grade liver dysplastic nodule specimens, 3 low-grade liver dysplastic nodule specimens, 50 liver cirrhosis tissue samples, and 20 normal liver tissue samples were measured using immunohistochemistry. The F-test, q-test, t-test, and chi-square test were used for statistical analysis of data. Results: ATAD2 was expressed in 56 HCC surgical specimens (93.33%), 35 HCC biopsy specimens (81.40%), and 2 high-grade liver dysplastic nodule specimens (2/2), but not in the low-grade liver dysplastic nodule, liver cirrhosis tissue, and normal liver tissue samples. The mean expression of ATAD2 was significantly higher in HCC tissues than in high-grade and low-grade liver dysplastic nodule tissues, liver cirrhosis tissue, and normal liver tissue (F = 22.96, q = 3.138, 3.972, 12.272, and 9.101, respectively, all P < 0.01). There were no significant differences in the mean expression and positive expression rate of ATAD2 between HCC surgical and biopsy specimens (t = 1.40, P > 0.05; χ² = 3.47, P >0.05). Of the 35 HCC biopsy specimens that expressed ATAD2, the mean ATAD2 expression was ≥1% in 35 specimens (100%), ≥3% in 27 specimens (77.14%), and ≥5 % in 23 specimens (65.71%). In addition, among the pathological grade I-II HCC biopsy specimens, the mean ATAD2 expression was ≥1% in 28 specimens (100%), ≥3% in 22 specimens (62.86%), and ≥5% in 19 specimens (54.29%). Moreover, ATAD2 expression in HCC was associated with serum alpha-fetoprotein level, presence of hepatitis B virus surface antigen (HBsAg), and presence of concurrent liver cirrhosis (t = 2.09, 2.30, and 2.18, respectively, all P < 0.05). Conclusion: ATAD2 may play an important role in HCC tumorigenesis, and may be involved in malignant transformation of cells. ATAD2 expression can be a valuable marker for differentiating the nature of lesions in liver biopsy tissues during clinical practice.

Distribution and Characteristics of Colletotrichum spp. Associated with Anthracnose of Strawberry in Hubei, China.

Anthracnose caused by Colletotrichum spp. is a serious disease of strawberry. The etiology of anthracnose of strawberry is complex, and several Colletotrichum spp. have been regarded as causal agents. In the present study, multilocus (actin, ß-tubulin, calmodulin, glyceraldehyde-3-phosphate dehydrogenase, and chitin synthase) phylogenetic analysis revealed that 100 isolates of Colletotrichum associated with anthracnose of strawberry in central China belong to five species. In total, 97 isolates were identified belonging to the Colletotrichum gloeosporioides species complex, with C. murrayae, C. gloeosporioides, C. fructicola, and C. aenigma accounting for 81, 8, 4, and 4% of the total isolates, respectively. Three isolates belonging to the C. acutatum complex were identified as C. nymphaeae. On inoculated strawberry plants, isolates of C. fructicola and C. murrayae species showed strong pathogenicity to both leaves and petioles of strawberry, with plant mortality 30 days after inoculation of 77.8 and 55.6%, respectively. C. gloeosporioides, C. aenigma, and C. nymphaeae showed strong pathogenicity to leaves but weak pathogenicity to petioles, with plant mortality 30 days after inoculation of 5.6, 16.7, and 11.1%, respectively. The five species were divided into four classes based on their maximum growth temperatures. Isolates of C. murrayae and C. gloeosporioides were more tolerant to high temperature (>34°C) than isolates of other species, followed by C. fructicola and C. aenigma. Isolates of C. nymphaeae, which are only distributed in areas of higher altitude (1,100 m), were highly sensitive to higher temperature. These results indicate that pathogenicity and adaptation to temperature are important factors in the distribution of Colletotrichum spp. on strawberry plants. This research may increase our understanding of how Colletotrichum spp. emerge and spread to geographical regions with different latitudes or elevations.

[Analysis of intrahepatic Glisson's sheath vascular disconnection approach for anatomical hepatectomy by three-dimensional laparoscope].

OBJECTIVE: To investigate the clinical value of the intrahepatic Glisson's sheath vascular disconnection approach for anatomical hepatectomy by three-dimensional(3D) laparoscope. METHODS: Twenty-two patients(11 liver cancer and 11 benign diseases)underwent hepatectomy in Zhujiang Hospital, Southern Medical University from November 2013 to March 2015 were involved. All patients were performed anatomical hepatectomy by 3D laparoscope with the intrahepatic Glisson's sheath vascular disconnection approach.The anatomical time of first hepatic portal, operation time, intraoperative bleeding, postoperation hospitalization and the complication incidences were observed. RESULTS: All the 22 cases were performed successfully with the intrahepatic Glisson's sheath vascular disconnection and the anatomic hepatectomy. The anatomical time of first hepatic portal, operation time, intraoperative bleeding, the postoperation hospitalization duration and complication incidence rate were(43±14) minutes(22-74 minutes), (295±89) minutes(105-480 minutes), (546±390) ml (50-1 500 ml), (9.8±2.8) days(5-16 days) and 18.2%(4/22)in 22 patients, respectively. There was no death. CONCLUSIONS: With the help of 3D laparoscope, the anatomy of Glisson pedicle could perform precisely and the dissection of vessels would be easy and accuracy. The 3D laparoscope is feasible for performing the anatomical hepatectomy.

High prevalence of vitamin D insufficiency in pregnant women working indoors and residing in Guiyang, China.

BACKGROUND/AIM: To assess the status of 25-hydroxyvitamin D [25(OH)D] in pregnant women in their second and third trimesters, who reside and work indoors in Guiyang, China. SUBJECTS AND METHODS: A total of 311 pregnant women in their 12th to 40th gestational week were engaged in employment located indoors in the urban area of Guiyang and completed a questionnaire on living habits. Levels of serum 25(OH)D were measured from fasting venous blood by liquid chromatography-mass spectrometry (LC-MS/MS). Levels of 25(OH)D were classified as vitamin D deficient [25(OH)D<20 ng/ml], insufficient [20 ng/ml≤25(OH)D<30 ng/ml], or sufficient [25(OH)D≥30 ng/ml]. RESULTS: The mean serum level of 25(OH)D was 14.69±6.81 ng/ml. Vitamin D deficiency, insufficiency and sufficiency were found in 260 (83.6%), 39 (12.5%), and 12 (3.9%) women, respectively. The mean level of 25(OH)D in the third trimester was significantly higher than in the second trimester (p<0.001). The mean 25(OH)D level in summer (June, July, August) was significantly higher than in the other seasons (p<0.001). The 25(OH)D level in pregnant women compliant with pre-natal calcium or multivitamin supplements was higher than in those not taking supplements (p<0.001). CONCLUSIONS: These results suggest that pregnant women residing in Guiyang urban area and working indoors are at high risk of vitamin D insufficiency, particularly during spring, winter, and autumn, regardless of use of pre-natal calcium and multivitamins. Appropriate vitamin D supplementation is necessary to improve maternal vitamin D nutrition.

Partial tolerance induced by transplantation of spatially separated thymuses: a cue for T cell retolerization in thymus grafts.

Thymus grafts made up of mixed syngeneic and xenogeneic thymus tissues could induce donor-specific tolerance to xenografts with no development of autoimmune syndrome (AIS). But the requirements for the simultaneous presentation of tissue antigens from both species in the process of T cell development in thymus grafts have not hitherto been defined. To do this, we setup a model in which xenothymus grafts from F344 rats were heterotopically implanted into BALB/c nude mice carrying syngeneic thymus grafts, and the grafts were either mixed together or spatially separately; next, we examined the induction of donor-specific tolerance, any pathological changes and the distribution of T lymphocytes. In contrast to the mixed thymus grafts, spatially separated thymus transplants could neither induce a long-term tolerance to skin grafts nor prevent AIS completely. (51) Cr-labelled cell-tracing experiments showed that mature peripheral T cells could re-enter into both kinds of thymus grafts, while the T cells isolated from the syngeneic thymus tended to concentrate in the xenothymus grafts. Hence, our data suggest that the immune tolerance induced by mixed thymus transplants could be partially reversed if the thymus tissues from donors and recipients were segregated by spatial telorism. The uneven recirculation of mature T cells might imply that T cells get retolerized within the thymus grafts.

Gene mapping and mutation screening in candidate genes in a Chinese family of autosomal dominant retinitis pigmentosa.

We wanted to find the gene defect in a Chinese pedigree with autosomal dominant form of retinitis pigmentosa (ADRP). A small Chinese family with retinitis pigmentosa was collected. The genetic analysis of the family suggested an autosomal dominant pattern. Microsatellite (STR) markers tightly linked to candidate genes for ADRP were selected for linkage analysis. We got a maximum LOD score of 0.87 between markers D19S210 and D19S418. Precursor mRNA-processing factor (PRPF) 31, 3, 8, rhodopsin (RHO), peripherin 2 (PRPH2 or RDS), rod outer segment protein 1 (ROM1), neural retina leucine zipper (NRL), cone-rod homeobox-containing (CRX), inosine-5-prime-monophosphate dehydrogenase, type I (IMPDH1) and retinitis pigmentosa 1 (RPI) were amplified by polymerase chain reaction (PCR) and screened by direct sequencing. One new sequence variation was found. It was the missence mutation c.148G > C (D50H) occurred in exon 1 of RDS gene which existed in all the effected individuals and one unaffected family member. The DNA sequence variation didn't cosegregate with the RP disease. We considered this transition was one new polymorphism which we speculate involved in the pathogenesis of ADRP and increased the risk of ADRP. Further study should be conducted to confirm the causative gene of this family.

[Influencing factors and their predictive value of skin graft survival after Meek grafting in severe burn patients].

Objective: To investigate the influencing factors and their predictive value of skin graft survival after Meek grafting in severe burn patients. Methods: A retrospective case-control study was conducted in 115 severe burn patients (95 males, 20 females, aged 1-74 years) who met the inclusion criteria and received Meek grafting in the First Affiliated Hospital of Army Medical University (the Third Military Medical University) from January 2013 to December 2019. The patients were divided into good skin graft survival group with skin graft survival rate≥70% (68 cases) and poor skin graft survival group with skin graft survival rate<70% (47 cases). The statistics of patients in the two groups were recorded during their first Meek grafting after admission including the gender, age, body mass index, full-thickness burn area, burn index, complication of inhalation injury, time from injury to operation, preoperative cystatin C level, preoperative albumin level, preoperative neutrophil, preoperative hemoglobin level, preoperative platelet count, and platelet count on the first, third, and fifth day after operation. The above indicators were statistically analyzed between the two groups with independent sample t test, Mann-Whitney U test, and chi-square test. A 1∶1 propensity score matching (PSM) of the gender, age, body mass index, full-thickness burn area, burn index, complication of inhalation injury, time from injury to operation of patients in the two groups were performed to eliminate the differences in baseline data, and then the above indicators of the remaining patients in the two groups were recorded and analyzed again. The indicators with statistically significant differences between the two groups after 1∶1 PSM were selected for multivariate logistic regression analysis to screen the independent risk factors affecting the skin graft survival after Meek grafting in severe burn patients. The receiver operating characteristic (ROC) curve of independent risk factors for predicting poor skin graft survival after Meek grafting in severe burn patients after 1∶1 PSM was drawn, and the area under the curve, the cut-off value, and the sensitivity and specificity under the cut-off value were calculated. The patients after 1∶1 PSM were divided into independent risk factor>the cut-off value group and independent risk factor≤the cut-off value group with the incidence of poor skin graft survival after Meek grafting compared using the chi-square test, and the relative risk of poor skin graft survival after Meek grafting was calculated. Results: Before 1∶1 PSM, there were no statistically significant differences in gender, age, body mass index, complication of inhalation injury, time from injury to operation, preoperative cystatin C level, preoperative albumin level, preoperative neutrophil, preoperative hemoglobin level of patients between the two groups (P>0.05); the full-thickness burn area and burn index of patients in poor skin graft survival group were significantly higher than those in good skin graft survival group (Z=-2.672, -2.882, P<0.01); the preoperative platelet count and the platelet count on the first, third, and fifth day after operation of patients in poor skin graft survival group were significantly lower than those in good skin graft survival group (Z=-3.411, -3.050, -2.748, -2.686, P<0.01). After 1∶1 PSM, 46 cases were remained in each group. There were no statistically significant differences in gender, age, body mass index, full-thickness burn area, burn index, complication of inhalation injury, time from injury to operation, preoperative cystatin C level, preoperative albumin level, preoperative neutrophil, preoperative hemoglobin level of remaining patients between the two groups (P>0.05); the preoperative platelet count and the platelet count on the first, third, and fifth day after operation of patients in poor skin graft survival group were significantly lower than those in good skin graft survival group (Z=-3.428, -2.940, t=-2.427, -2.316, P<0.05 or P<0.01). Multivariate logistic regression analysis showed that the preoperative platelet count was the only independent risk factor affecting the skin graft survival after Meek grafting in severe burn patients (odds ratio=0.994, 95% confidence interval=0.989-0.998, P<0.01). The area under the ROC curve of preoperative platelet count predicting poor skin graft survival after Meek grafting in 92 patients was 0.707 (95% confidence interval=0.603-0.798, P<0.01), and the cut-off value of preoperative platelet count was 98×109/L, with sensitivity of 54.3% and specificity of 78.3% under the cut-off value. The incidence of poor skin survival after Meek grafting of patients in preoperative platelet count≤98×109/L group was 71.4% (25/35), which was obviously higher than 36.8% (21/57) in preoperative platelet count>98×109/L group (χ2=10.376, P<0.01). Compared with that in preoperative platelet count>98×109/L group, patients in preoperative platelet count≤98×109/L group had a relative risk of poor skin graft survival after Meek grafting of 2.211 (95% confidence interval=1.263-3.870). Conclusions: Preoperative platelet count is an independent risk factor affecting the skin graft survival after Meek grafting in severe burn patients and has a good predictive value. Meek grafting should be performed with caution when the preoperative platelet count of patients is≤98×109/L.

[Clinical application of extracorporeal membrane oxygenation in the treatment of burn patients with acute respiratory distress syndrome: a retrospective analysis and systematic review].

Objective: To analyze the clinical effect of extracorporeal membrane oxygenation (ECMO) in the treatment of burn patients with acute respiratory distress syndrome (ARDS). Methods: The retrospective observational study and the systematic review were applied. From March 2014 to July 2020, five burn patients with ARDS received ECMO treatment in the First Affiliated Hospital of Army Medical University (the Third Military Medical University). All the five patients were male, aged from 40 to 62 years. The average total burn surface area was 58.8% total body surface area (TBSA) and four cases had severe inhalation injury. Patient's ECMO starting time, duration and mode, and whether successfully weaned or the cause of death, and others. were recorded. Furthermore, the changes of oxygenation and infection before, during, and after utilizing ECMO were analyzed. PubMed and Web of Science from the establishment of each database to August 2021 were searched using "Extracorporeal Membrane Oxygenation", "ECMO", "burn", "inhalation" as the search terms and "Title/Abstract" as the field to retrieve the clinical articles that meet the selection criteria . Basic information were extracted from the articles, including sample size, gender, age, total burn area, inhalation injury, the indication of ECMO, the start and lasting time of ECMO, ECMO mode, rate of successful weaning, complications of ECMO, mortality, the combined application of continuous renal replacement therapy (CRRT). Results: Five patients started venovenous ECMO on an average of 10.2 days after injury and lasted an average of 180.4 hours. Three out of 5 patients were weaned successfully with one patient survived. Four patients died of multiple organ dysfunction syndrome (MODS) and septic shock. Compared with those before ECMO treatment, the arterial oxygen partial pressure (PaO2) and oxygen saturation in arterial blood (SaO2) of three successfully weaned patients obviously increased during and after ECMO treatment. The fraction of inspired oxygen (FiO2) decreased below 50% and PaO2/FiO2 ratio increased above 200 mmHg (1 mmHg=0.133 kPa) during and after ECMO. Furthermore, lactic acid and respiratory rate decreased, basically. Compared with those before ECMO, PaO2 and SaO2 in the other two patients during ECMO, who failed to be weaned, continuously decreased while lactic acid increased. Before and during ECMO, the PaO2/FiO2 ratios of unsuccessfullg weaned cases were less than 200 mmHg, and partial pressure of carbon dioxide in arterial blood (PaCO2) were more than 40 mmHg. Compared with those before ECMO, there were no significant changes in body temperature during and after ECMO, which were less than 38 ℃. Compared with those before ECMO, the leucocyte number (the index without this in unsuccessfully weaned cases was omitted, the same as below) in four patients showed a significant decrease during ECMO, but rose after removal of ECMO. The proportion of neutrophils in three patients were slightly higher during ECMO than before ECMO, and did not change significantly after removal of ECMO. Compared with those before ECMO, platelet counts in three patients were significantly reduced during ECMO, and all five patients during ECMO were below normal levels. Compared with those before ECMO, the procalcitonin levels in four deaths were significantly increased during ECMO. Catheter culture of microorganism was performed in three successfully weaned patients, all of which were negative. A total of 13 literature were included, ranging from 1990 to 2019. The sample size in 6 studies was less than 10, and the sample size in 4 studies was between 10 and 20, and only 2 literatures had a sample size larger than 50. ECMO was applied in 295 burn patients with overall mortality of 48.8% (144/295), including 157 adults and 138 children. The most common indication of ECMO was severe ARDS. Among 157 adult burn patients (95 males and 65 females), 36 cases had inhalation injury. The average burn area was 27%-37%TBSA in 5 reported studies and was more than 50%TBSA in 2 reported studies. The most common mode was venovenous ECMO. ECMO treatment began 26.5 hours to 7.4 days after injury and lasted from 90 hours to 18 days, and the rate of successful weaning ranged from 50% to 100%. The most common complications were bleeding and infection. The mortality was 52.9% (83/157). MODS and sepsis were the leading causes of death. Among 138 pediatric burn patients (77 boys and 61 girls), 29 patients had inhalation injury. The average burn area was 17%-50.2%TBSA in 3 studies. ECMO treatment lasted from 165.2 hours to 324.4 hours. Bleeding was the most common complication. The mortality was 44.2% (61/138). Conclusions: ECMO is an effective strategy for the salvage treatment of burns complicated with ARDS. Furthermore, the prevention and treatment of bleeding, infection and organ dysfunction should be emphasized during the use of ECMO. More importantly, evidence-based guidelines for burns are urgently needed to further improve the clinical effect of ECMO.

[Effects and mechanism of mitochondrial transcription factor A and cytochrome c oxidase pathway in the energy production of hypoxic cardiomyocytes of rats regulated by tumor necrosis factor receptor associated protein 1].

Objective: To investigate the effects and mechanism of mitochondrial transcription factor A (TFAM) and cytochrome c oxidase (COX) pathway in the energy production of hypoxic cardiomyocytes of rats regulated by tumor necrosis factor receptor associated protein 1 (TRAP1). Methods: The cardiomyocytes were isolated from 135 neonatal Sprague-Dawley rats (aged 1-3 d) and cultured for the following experiments. (1) Cells were collected and divided into normoxia blank control (NBC) group, hypoxia blank control (HBC) group, hypoxia+ TRAP1 over-expression control (HTOC) group, and hypoxia+ TRAP1 over-expression (HTO) group according to the random number table (the same grouping method below), with 1 bottle in each group. Cells in NBC group were cultured routinely, cells in HBC group were cultured in hypoxic condition for 6 hours after routine culture, cells in HTOC and HTO groups were respectively added with TRAP1 over-expression empty virus vector and TRAP1 over-expression adenovirus vector virus suspension for transfection for 48 hours after routine culture and then cultured in hypoxic condition for 6 hours. The protein expression of TFAM of cells in each group was detected by Western blotting. (2) Cells were collected and divided into NBC, HBC, HTOC, HTO, HTO+ TFAM interference control (HTOTIC), and HTO+ TFAM interference (HTOTI) groups, with 1 well in each group. Cells in the former 4 groups were dealt with the same methods as the corresponding groups in experiment (1). Cells in HTOTIC and HTOTI groups were respectively added with TFAM interference empty virus vector and TFAM interference adenovirus vector virus suspension for transfection for 48 hours, and the other processing methods were the same as those in HTO group. The content of ATP of cells in each group was determined by ATP determination kit and microplate reader, and the COX activity of cells in each group was determined by COX activity assay kit and microplate reader. (3) Cells were collected and divided into NBC group, normoxia+ sodium azide (NSA) group, HBC group, and hypoxia+ sodium azide (HSA) group, with 1 well in each group. Cells in NBC and HBC groups were respectively dealt with the same methods as the corresponding groups in experiment (1). Cells in NSA and HSA groups were respectively added with 32 nmol sodium azide at 30 min before experiment or hypoxia, and then cells in HSA group were cultured in hypoxic condition for 6 hours. The content of ATP was determined by the same method as above. The above three experiments were repeated for three times. Data were statistically analyzed with one-way analysis of variance and least significant difference test. Results: (1) Compared with that in NBC group, the protein expression of TFAM of cells in HBC group was significantly decreased (P<0.01). Compared with that in HBC group or HTOC group, the protein expression of TFAM of cells in HTO group was significantly increased (P<0.01). (2) Compared with 0.552±0.041 and 1.99±0.15 in NBC group, the COX activity (0.270±0.044) and ATP content (1.09±0.11) of cells in HBC group were significantly decreased (P<0.01). Compared with 0.269±0.042 and 1.17±0.12 in HBC group and those in HTOC group, the COX activity (0.412±0.032 and 0.404±0.016) and ATP content (1.75±0.06 and 1.69±0.07) of cells in HTO and HTOTIC groups were significantly increased (P<0.01). Compared with those in HTO and HTOTIC groups, the COX activity (0.261±0.036) and ATP content (1.23±0.07) of cells in HTOTI group were significantly decreased (P<0.01). (3) Compared with that in NBC group, the ATP content of cells in NSA and NBC groups was significantly decreased (P<0.01). Compared with that in HBC group, the ATP content of cells in HSA group was significantly decreased (P<0.01). Conclusions: TRAP1 can increase the COX activity of cardiomyocytes by raising the expression of TFAM, and finally alleviate the impairment in energy production of cardiomyocytes caused by hypoxia.

[Recommendations for normalizing the medical practices of burn treatment during the outbreak of coronavirus disease 2019].

2019 novel coronavirus (2019-nCoV) is one of the beta coronaviruses and is identified as the pathogen of the severe " coronavirus disease 2019 (COVID-19)" in 2019. China manages COVID-19 according to the reguirement of the highest level infectious diseases in China. Currently, the prevention and control of COVID-19 in China is at a critical period. Burn Department as an emergency discipline is confronted with risk of 2019-nCoV infection. Based on the guidelines for the diagnosis and treatment of COVID-19 (6th trial edition), in combination with the latest literature at home and abroad, the features of the COVID-19, the recommendations for the COVID-19 prevention and control issued by the National Health Commission of China, and the management experience of COVID-19 diagnosis and treatment of other related disciplines, we put forward some recommendations for the medical practices of burn treatment during the outbreak of the COVID-19 in outpatient and emergency, inpatient treatment, and the management of operation theatres and wards, etc. We hope these recommendations could benefit the medical professionals in the field of burn treatment and relevant hospital management during the outbreak of COVID-19, improve burn treatment, and avoid or reduce the risk of infection of medical staff.

[Retrospective study on the characteristics of early organ injury in elderly patients with severe burns].

Objective: To analyze the clinical characteristics of early organ injury in elderly patients with severe burns and the effects on the prognosis of patients. Methods: From January 2010 to August 2018, 62 patients with severe burns (43 men and 19 women, aged from 60 to 89 years at the time of admission) who were hospitalized in the Institute of Burn Research of the First Affiliated Hospital of Army Medical University (the Third Military Medical University, hereinafter referred to as the author's affiliation), meeting the inclusion criteria, were included in elderly (E) group, and 124 patients with severe burns (86 men and 38 women, aged from 18 to 59 years at the time of admission) at the same term were included in young and middle-aged (YM) group. Treatment of patients in the 2 groups followed the conventional procedures of the author's affiliation. The following data of patients in the 2 groups were retrospectively analyzed. (1) Fluid replacement volume and urine volume within the first and second post injury hour (PIH) 24 were recorded. The levels of hemoglobin, haematocrit, and blood lactic acid at admission, PIH 24 and 48 were recorded. (2) The creatine kinase isozyme-MB (CK-MB), total bilirubin, blood creatinine, oxygenation index, and blood platelet count at admission, at shock stage, and on post injury day (PID) 3 to 7 were collected. (3) The days of seriously or critically ill and deaths were recorded. Data were processed with chi-square test, group t test, Mann-Whitney U test, analysis of variance for repeated measurement, and Bonferroni correction. Results: (1) There were no statistically significant differences in fluid replacement volume within the first and second PIH 24, and urine volume within the second PIH 24 between patients in the 2 groups (t=0.351, 1.307, 1.110, P>0.05). The urine volume of patients in group E within the first PIH 24 was significantly less than that in group YM (t=5.628, P<0.05). There were no statistically significant differences in levels of hemoglobin (t=0.011, 1.075, 0.239), haematocrit (t=0, 0.033, 0.199), and blood lactic acid (t=0.017, 1.002, 0.739) at admission, PIH 24 and 48 between patients in the 2 groups (P>0.05). (2) There were no statistically significant differences in levels of CK-MB at admission and on PID 3 to 7 between patients in the 2 groups (t=0.069, 0.001, P>0.05). The level of CK-MB of patients in group E at shock stage was significantly higher than that in group YM (t=4.017, P<0.05). There were no statistically significant differences in levels of total bilirubin at admission and on PID 3 to 7 between patients in the 2 groups (t=0.227, 0.002, P>0.05). However, the level of total bilirubin of patients in group E at shock stage was significantly higher than that in group YM (t=6.485, P<0.05). The levels of blood creatinine of patients in group E at admission and shock stage were significantly higher than those in group YM (t=4.226, 12.299, P<0.05 or P<0.01), while there was no statistically significant difference between them on PID 3 to 7 (t=0.693, P>0.05). The oxygenation indexes of patients in group E at admission and shock stage and on PID 3 to 7 [(371±16), (263±16), and (228±18) mmHg (1 mmHg=0.133 kPa)] were lower than (420±13), (327±13), and (281±17) mmHg of patients in group YM, respectively (t=5.650, 9.782, 4.856, P<0.05 or P<0.01). There were no statistically significant differences in levels of blood platelet count at admission and shock stage between patients in the 2 groups (t=0.038, 0.588, P>0.05), while the level of blood platelet count of patients in group E on PID 3 to 7 was significantly lower than that in group YM (t=6.636, P<0.05). (3) The days of seriously or critically ill and death rate of patients in group E were respectively longer or higher than those in group YM (Z=-2.303, χ(2)=13.676, P<0.05 or P<0.01). Conclusions: In the case of the same tissue perfusion at shock stage, injuries in heart, liver, kidney, lung, and coagulation system in elderly patients with severe burns are more obvious than those in young and middle-aged patients, with more severe illness and higher mortality.

[In vitro study of effects of transient receptor potential vanilloid 1 on autophagy in early hypoxic mouse cardiomyocytes and the mechanism].

Objective: To explore the effects of transient receptor potential vanilloid 1 (TRPV1) on autophagy in early hypoxic mouse cardiomyocytes and the mechanism in vitro. Methods: The hearts of 120 C57BL/6 mice aged 1-2 days, no matter male or female, were isolated, and then primary cardiomyocytes were cultured and used for the following experiments, the random number table was used for grouping. (1) The cells were divided into normoxia group and hypoxia 3, 6, and 9 h groups, with one well in each group. The cells in normoxia group were routinely cultured (the same below), the cells in hypoxia 3, 6, and 9 h groups were treated with fetal bovine serum-free and glucose-free Dulbecco' s modified Eagle medium under low oxygen condition in a volume fraction of 1% oxygen, 5% carbon dioxide, and 94% nitrogen for 3, 6, and 9 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1, TRPV1 were determined with Western botting. (2) The cells were divided into normoxia group and hypoxia group, with two coverslips in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above. The positive expression of TRPV1 was detected by immunofluorescence assay. (3) The cells were divided into 4 groups, with one well in each group. The cells in simple hypoxia group were treated with hypoxia for 6 h as above, and the cells in hypoxia+ 0.1 µmol/L capsaicin group, hypoxia+ 1.0 µmol/L capsaicin group, and hypoxia+ 10.0 µmol/L capsaicin group were respectively treated with 0.1, 1.0, 10.0 µmol/L capsaicin for 30 min before hypoxia for 6 h. The protein expressions of LC3, Beclin-1, and TRPV1 were detected by Western blotting. (4) The cells were divided into 5 groups, with 5 wells in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ chloroquine group, hypoxia+ capsaicin group, and hypoxia+ capsaicin+ chloroquine group were treated with hypoxia for 6 h after being cultured with 50 µmol/L chloroquine, 10.0 µmol/L capsaicin, and 50 µmol/L chloroquine+ 10.0 µmol/L capsaicin for 30 min, respectively. Viability of cells was detected by cell counting kit 8 assay. (5) The cells were divided into simple hypoxia group and hypoxia+ 10.0 µmol/L capsaicin group, with one well in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ 10.0 µmol/L capsaicin group were treated with 10.0 µmol/L capsaicin for 30 minutes and then with hypoxia for 6 h. The protein expressions of lysosomal associated membrane protein 1 (LAMP-1) and LAMP-2 were detected by Western blotting. Each experiment was repeated for 3 or 5 times. Data were processed with one-way analysis of variance, least significant difference t test, and Bonferroni correction. Results: (1) Compared with those of normoxia group, the protein expressions of LC3, Beclin-1, and TRPV1 were significantly increased in cardiomyocytes of hypoxia 3, 6, and 9 h groups (t(3 h)=4.891, 5.890, 4.928; t(6 h)=9.790, 6.750, 10.590; t(9 h)=6.948, 6.764, 5.049, P<0.05 or P<0.01), which of hypoxia 6 h group were the highest (1.08±0.05, 1.12±0.10, 0.953±0.071, respectively). (2) The density of TRPV1 in cell membrane and inside the cardiomyocytes in hypoxia group was significantly increased with lump-like distribution, and the expression of TRPV1 was higher than that in normoxia group. (3) Compared with those of simple hypoxia group, the protein expression of Beclin-1 in cardiomyocytes of hypoxia+ 0.1 µmol/L capsaicin group was increased (t=10.488, P<0.01), while the protein expressions of LC3 and TRPV1 were increased without statistically significant differences (t=4.372, 3.026, P>0.05); the protein expressions of LC3, TRPV1, and Beclin-1 in cardiomyocytes of hypoxia+ 1.0 µmol/L capsaicin group and hypoxia+ 10.0 µmol/L capsaicin group were significantly increased (t=15.505, 5.773, 13.430; 20.915, 8.054, 16.384; P<0.05 or P<0.01), which of hypoxia+ 10.0 µmol/L capsaicin group were the highest (2.33±0.09, 1.34±0.07, 1.246±0.053, respectively). (4) Compared with 0.585±0.045 in normoxia group, the cardiomyocyte viability in hypoxia group was significantly decreased (0.471±0.037, t=4.365, P<0.05). Compared with that in hypoxia group, the cardiomyocyte viability in hypoxia+ chloroquine group was further decreased (0.350±0.023, t=6.216, P<0.01), while 0.564±0.047 in hypoxia+ capsaicin group was significantly increased (t=3.489, P<0.05). Compared with that in hypoxia+ chloroquine group, the cardiomyocyte viability in hypoxia+ capsaicin+ chloroquine group did not significantly change (0.364±0.050, t=0.545, P>0.05). (5) Compared with 0.99±0.04 and 0.54±0.04 in simple hypoxia group, the protein expressions of LAMP-1 and LAMP-2 in hypoxia+ 10.0 µmol/L capsaicin group were significantly increased (1.49±0.06, 0.81±0.05, t=12.550, 7.442, P<0.01). Conclusions: TRPV1 can further promote the expression of autophagy-related proteins in hypoxic cardiomyocytes through autophagy-lysosomal pathway, enhance autophagy activity, and improve autophagic flow for alleviating early hypoxic cardiomyocyte injury.

A combination of donor specific transfusion and rapamycin prolongs cardiac allograft survival in mice.

BACKGROUND: We sought to investigate the effects of donor-specific transfusion (DST) combined with rapamycin (RPM) on engraftment and demonstrate the mechanisms. METHODS: B6 was treated with DST (on day -8) combined with RPM (from day -7 to -1, 1.5 microL/g per day). On day 0, B6 splenocytes were harvested to coculture with Balb/c splenocytes pretreated with mitomycin C in 1-way mixed lymphocyte cultures (MLC). After 48 hours, the proliferation, we measured apoptosis and the CD4(+)CD25(+) T-cell ratio of cultured cells. The heterotopic cardiac transplantation model was performed from Balb/c to B6 using the recipients pretreated with this protocol. On day 0, RPM was withdrawn and the transplantations performed. The mean survival time (MST) of the grafts evaluated and in vitro assays measured. RESULTS: Both in vitro and in vivo, the protocol suppressed the proliferation of splenocytes as well as enhanced apoptosis and the CD4(+)CD25(+) T-cell ratio. The MST of the grafts was 35 +/- 14.4 days. CONCLUSIONS: Our study showed that DST combined with RPM prolonged cardiac allograft survival even if we withdrew immunosuppressants after transplantation. This result may be associated with inhibition of T-cell proliferation. We also demonstrated that RPM expanded CD4(+)CD25(+) T cells and increased splenocyte apoptosis.

[Mechanism of protective effects of tumor necrosis factor receptor associated protein 1 on hypoxic cardiomyocytes of rats].

Objective: To investigate the mechanism of protective effects of tumor necrosis factor receptor associated protein 1 (TRAP1) on hypoxic cardiomyocytes of rats. Methods: Primary cultured cardiomyocytes were obtained from neonatal Sprague-Dawley rats (aged 1 to 3 days) and then used in the following experiments. (1) Cells were divided into group TRAP1 and control group according to the random number table (the same grouping method below), and then the total protein of cells was extracted. Total protein of cells in group TRAP1 was added with mouse anti-rat TRAP1 monoclonal antibody, while that in control group was added with the same type of IgG from mouse. Co-immunoprecipitation and protein mass spectrography analysis were used to determine the possible proteins interacted with TRAP1. (2) Cells were divided into normoxia blank control group (NBC), normoxia+ TRAP1 interference control group (NTIC), normoxia+ TRAP1 interference group (NTI), normoxia+ TRAP1 over-expression control group (NTOC), and normoxia+ TRAP1 over-expression group (NTO), with 1 well in each group. Cells in group NBC were routinely cultured, while cells in the latter four groups were respectively added with TRAP1 RNA interference empty virus vector, TRAP1 RNA interference adenovirus vector, TRAP1 over-expression empty virus vector, and TRAP1 over-expression adenovirus vector. Another batch of cells were divided into group NBC, hypoxic blank control group (HBC), hypoxic+ TRAP1 interference control group (HTIC), hypoxic+ TRAP1 interference group (HTI), hypoxic+ TRAP1 over-expression control group (HTOC), and hypoxic+ TRAP1 over-expression group (HTO), with 1 well in each group. Cells in hypoxic groups were under hypoxic condition for 6 hours after being treated as those in the corresponding normoxia groups, respectively. The mRNA expression of cytochrome c oxidase subunit Ⅱ (COXⅡ) of cells in each group was detected by real time fluorescent quantitive reverse transcription polymerase chain reaction. Experiments were repeated for three times. (3) Cells were divided into group NBC, group HBC, group HTOC, group HTO, hypoxic+ TRAP1 over-expression+ COXⅡinterference control group (HTOCIC), and hypoxic+ TRAP1 over-expression+ COXⅡinterference group (HTOCI), with 3 wells in each group. Cells in the previous 4 groups were treated as those in experiment (2). Cells in group HTOCIC and HTOCI were respectively transfected with COXⅡ RNA interference empty virus vector and COXⅡ RNA interference adenovirus vector, and then both added with TRAP1 over-expression adenovirus vector. The proliferation activity of cells was determined by cell counting kit 8 and microplate reader, and the ratio of death cells was measured by propidium lodide and Hoechst 33342 staining. Another batch of cells were divided into group NBC, group HBC, group HTIC, group HTI, hypoxic+ TRAP1 interference+ COXⅡover-expression control group (HTICOC), and hypoxic+ TRAP1 interference+ COXⅡ over-expression group (HTICO), with 3 wells in each group. Cells in the previous 4 groups were treated as those in experiment (2). Cells in group HTICOC and HTICO were both transfected with TRAP1 RNA interference adenovirus vector, and then respectively added with COXⅡ over-expression empty virus vector and COXⅡ over-expression adenovirus vector. The proliferation activity of cells and the ratio of death cells were detected as before. Experiments were repeated for three times. Data were processed with one-way analysis of variance and LSD test. Results: (1) The expression of TRAP1 was found in cells of group TRAP1, while that was not found in cells of control group. The possible proteins interacted with TRAP1 were keratin, COXⅡ, and an unknown protein with predicted molecular weight 13×103. (2) Compared with that in group NBC, the mRNA expression of COXⅡof cells had no significant change in group NTIC and group NTOC (with P values above 0.05), but significantly decreased in group NTI (P<0.01), and significantly increased in group NTO (P<0.01). Compared with that in group NBC, the mRNA expression of COXⅡof cells in group HBC was significantly decreased (P<0.01). Compared with that in group HBC, the mRNA expression of COXⅡof cells had no significant change in group HTIC and group HTOC (with P values above 0.05), but significantly decreased in group HTI (P<0.01), and significantly increased in group HTO (P<0.01). (3) The proliferation activity of cells in group NBC, group HBC, group HTOC, group HTO, group HTOCIC, and group HTOCI was respectively 0.498±0.022, 0.303±0.018, 0.313±0.032, 0.456±0.031, 0.448±0.034, and 0.335±0.026, and the ratios of death cells in above groups were respectively (4.7±1.5)%, (24.7±3.1)%, (26.0±2.7)%, (13.3±2.5)%, (12.7±2.1)%, and (21.0±1.7)%. Compared with those in group NBC, the proliferation activity of cells in HBC was decreased, while the ratio of death cells was increased (with P values below 0.01). Compared with those in group HBC, the proliferation activity of cells and the ratio of death cells in group HTOC had no significant change (with P values above 0.05), while the proliferation activity of cells was increased and the ratio of death cells was decreased in group HTO (with P values below 0.01). Compared with those in group HTO, the proliferation activity of cells and the ratio of death cells in group HTOCIC had no significant change (with P values above 0.05), while the proliferation activity of cells was decreased and the ratio of death cells was increased in group HTOCI (with P values below 0.01). (4) The proliferation activity of cells in group NBC, group HBC, group HTIC, group HTI, group HTICOC, and group HTICO was respectively 0.444±0.025, 0.275±0.016, 0.283±0.021, 0.150±0.009, 0.135±0.011, and 0.237±0.017, and the ratios of death cells in above groups were respectively (3.7±0.6)%, (21.0±2.7)%, (20.3±3.1)%, (31.7±2.5)%, (33.3±3.2)%, and (19.3±1.5)%. Compared with those in group HBC, the proliferation activity of cells and the ratio of death cells in group HTIC had no significant change (with P values above 0.05). Compared with those in group HBC and group HTIC, the proliferation activity of cells was decreased and the ratio of death cells was significantly increased in group HTI (with P values below 0.01). Compared with those in group HTI, the proliferation activity of cells and the ratio of death cells in group HTICOC had no significant change (with P values above 0.05), while the proliferation activity of cells was increased and the ratio of death cells was significantly decreased in group HTICO (with P values below 0.01). Conclusions: TRAP1 can up-regulate the expression of COXⅡ mRNA, and COXⅡ is one of the downstream effector molecules that TRAP1 mediates its protective effects on hypoxic cardiomyocytes.