[Therapeutic strategy for totally thoracoscopic repeat mitral valve surgery under moderate hypothermia-induced ventricular fibrillation].

Objective: To examine the therapeutic strategy and its impacting factors by analyzing the perioperative outcomes of total thoracoscopic repeat mitral valve surgery under moderate hypothermia-induced ventricular fibrillation with cardiopulmonary bypass. Methods: This study is a retrospective case series. Totally 63 patients who underwent repeat mitral valve surgery by the same surgeon from January 2021 to December 2023 in Department of Cardiovascular Surgery, the First Medical Center of People's Liberation Army General Hospital were retrospectively enrolled. There were 28 males and 35 females with an age of (58.3±15.9) years (range: 13 to 84 years). Surgery was performed using a totally thoracoscopic approach under moderate hypothermia-induced ventricular fibrillation. Mitral valvuloplasty was completed in 32 cases and mitral valve replacement in 31 cases. Preoperative baseline data and perioperative outcomes of the patients were collected and Logistic regression was used to analyze independent influencing factors of premature ventricular contractions in the early postoperative period. Results: The intraoperative cardiopulmonary bypass time was (191.5±50.9) minutes (range: 95 to 286 minutes), and the hypothermic ventricular fibrillation time was (99.0±39.8) minutes (range: 34 to 203 minutes). The anal temperature before the start of cardiopulmonary bypass was (36.3±0.5) ℃ (range: 35.2 to 38.0 ℃), the lowest intraoperative anal temperature was (27.3±1.3) ℃(range: 23.7 to 30.1 ℃), and the anal temperature at the time of the cessation of cardiopulmonary bypass was (36.3±0.4) ℃ (range: 35.2 to 37.0 ℃), and excessive rewarming was observed in 33 cases. Six cases applied the artificial heart assist device. Seventeen cases developed premature ventricular contractions in the early postoperative period. Two cases developed neurologic complications. Five cases developed respiratory complications. One case developed urological systemic complications. Six cases were mechanically ventilated for more than 3 days, and the duration of ICU stay in 16 cases was more than 3 days and the postoperative discharge time of (M(IQR)) 8.0 (3.5) days (range: 3 to 26 days). Two cases died or were discharged voluntarily. Logistic regression results showed that persistent preoperative atrial fibrillation (OR=11.424, 95%CI: 1.477 to 144.564, P=0.033) and excessive rewarming (OR=15.249, 95%CI: 1.357 to 279.571, P=0.038) were independent risk factors for the appearance of premature ventricular contractions in the early postoperative period. Conclusions: The technique of total thoracoscopic surgery under induced moderate hypothermic ventricular fibrillation with cardiopulmonary bypass could be applied to repeated mitral valve surgeries with less trauma and faster recovery. Persistent preoperative atrial fibrillation and excessive rewarming are independent risk factors for the occurrence of premature ventricular contractions in the early postoperative period.

Endometrioid adenocarcinoma: combined multiparametric MRI and tumour marker HE4 to evaluate tumour grade and lymphovascular space invasion.

AIM: To assess the value of semi-quantitative dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) and quantitative diffusion-weighted imaging parameters combined with human epididymis protein 4 (HE4) in predicting the pathological grade and lymphovascular space invasion (LVSI) of endometrioid adenocarcinoma (EAC). MATERIALS AND METHODS: Between October 2018 and December 2021, 60 women (mean age, 55 [range, 32-77] years) with EAC underwent preoperative pelvic MRI and HE4 level measurements. The positive enhancement integral (PEI), time to peak, maximum slope of increase (MSI), and maximum slope of decrease were measured by manually drawing a region of interest on the neoplastic tissue. The receiver operating characteristic curve was used to calculate the diagnostic efficiency of the single parameter and combined factors. RESULTS: Lower apparent diffusion coefficients (ADCs) were observed in high-grade tumours (G3) than in low-grade tumours (G1/G2). PEI, MSI, and HE4 levels were higher in the high-grade tumours than in the low-grade tumours (p<0.05). The area under the curve (AUC) for G3 diagnosis using multiparametric MRI combined with HE4 was 0.929. ADC values were significantly lower in the EAC with LVSI than in those without LVSI. Tumours with LVSI showed higher PEI and HE4 levels than those without LVSI (p<0.05). The AUC for LVSI-positive diagnosis using multiparametric MRI combined with HE4 was 0.814. CONCLUSION: Semi-quantitative DCE-MRI, ADC values, and serum HE4 levels can be used to predict tumour grade and LVSI, and the prediction efficiency of multiparametric MRI combined with serum HE4 is better than that of any single factor.

[BCS1Neonatal growth retardation and lactic acidosis initiated by novel mutation sites in L gene].

This study aims to analyze the clinical characteristics and genetic variations of two cases with developmental delay and lactic acidosis in a family, and to explore the relationship between genetic variations and clinical features. A retrospective analysis was conducted on the clinical characteristics of two siblings with developmental delay and lactic acidosis who were treated at the Neonatal Department of Children's Hospital of Chongqing Medical University in May 2019 and December 2021, respectively. Whole-exome sequencing was used to detect genetic variations in the affected children. Homology modeling of the BCS1L protein was performed to analyze the structural and functional changes of the protein. The correlation between genetic variations and clinical phenotypes was analyzed. The results showed that the main clinical features of the two affected children in this family were manifestations of mitochondrial respiratory chain complex Ⅲ deficiency, including prematurity, developmental delay, respiratory failure, lactic acidosis, cholestasis, liver dysfunction, renal tubular lesions, coagulation dysfunction, anemia, hypoglycemia, hypotonia, and early death. Whole-exome sequencing revealed a novel deletion mutation c.486_488delGGA (p.E163del) and a novel missense mutation c.992C>T (p.T331I) in the BCS1L gene. Structural analysis of the homology modeling showed that the compound heterozygous mutation had a significant impact on protein function. In conclusion, the novel mutation site c.992C>T (p.T331I) in the BCS1L gene is a "likely pathogenic" mutation, and the compound heterozygous mutation is closely related to the phenotype of mitochondrial respiratory chain complex Ⅲ deficiency.

[Clinical effect of laparoscopy for the treatment of biliary stricture after biliary dilatation operation].

Objective: To investigate the safety and effect of laparoscopy for the treatment of biliary stricture after the biliary dilatation operation. Methods: The clinical data of 78 patients,including 27 males and 51 females aged (48.6±14.2)years(range:17 to 76 years),who presented biliary stricture after biliary dilatation operation from January 2017 to June 2021 in the Department of Minimally Invasive Hepatobiliary Surgery,Hunan Provincial People's Hospital,were retrospectively collected,with 38 cases in the laparoscopy group and 40 cases in the laparotomy group. Of the 78 patients,there were 67 cases of cholangiojejunostomy stricture and 11 cases of stricture of the high intrahepatic bile duct. Statistical methods such as t-test and χ2 test were carried out to compare perioperative clinical data and follow-up information between the two groups. Results: Less intraoperative blood loss((102.6±76.4)ml vs. (162.5±105.9) ml, t=-2.874,P=0.005),shorter postoperative stay length of stay((10.5±3.7)days vs. (14.5±6.4)days, t=-3.379,P=0.001) and shorter waiting time for postoperative anal exhaust((2.0±0.6)days vs. (2.5±0.9)days, t=-2.827,P=0.006) were found in the laparoscopy group than that in the laparotomy group,with statistically significant differences. While there was no statistically difference in the operative time((252.8±54.7)minutes vs. (257.4±68.6)minutes,t=-0.331,P=0.742). Postoperative review and follow-up did not show statistically significant differences between the two groups in the residual stone rate(5.3%(2/38) vs. 5.0%(2/40)) and the incidence of recurrent biliary stricture(5.3%(2/38) vs. 7.5%(3/40))(both P>0.05). Conclusion: Laparoscopy may be safe and effective in the treatment of biliary stricture after the biliary dilatation operation,with less trauma,faster recovery compared to laparotomy.

[Overexpression of NAT10 induced platinum drugs resistance in breast cancer cell].

Objective: To observe the platinum drugs resistance effect of N-acetyltransferase 10 (NAT10) overexpression in breast cancer cell line and elucidate the underlining mechanisms. Methods: The experiment was divided into wild-type (MCF-7 wild-type cells without any treatment) group, NAT10 overexpression group (H-NAT10 plasmid transfected into MCF-7 cells) and NAT10 knockdown group (SH-NAT10 plasmid transfected into MCF-7 cells). The invasion was detected by Transwell array, the interaction between NAT10 and PARP1 was detected by co-immunoprecipitation. The impact of NAT10 overexpression or knockdown on the acetylation level of PARP1 and its half-life was also determined. Immunostaining and IP array were used to detect the recruitment of DNA damage repair protein by acetylated PARP1. Flow cytometry was used to detect the cell apoptosis. Results: Transwell invasion assay showed that the number of cell invasion was 483.00±46.90 in the NAT10 overexpression group, 469.00±40.50 in the NAT10 knockdown group, and 445.00±35.50 in the MCF-7 wild-type cells, and the differences were not statistically significant (P>0.05). In the presence of 10 µmol/L oxaliplatin, the number of cell invasion was 502.00±45.60 in the NAT10 overexpression group and 105.00±20.50 in the NAT10 knockdown group, both statistically significant (P<0.05) compared with 219.00±31.50 in wild-type cells. In the presence of 10 µmol/L oxaliplatin, NAT10 overexpression enhanced the binding of PARP1 to NAT10 compared with wild-type cells, whereas the use of the NAT10 inhibitor Remodelin inhibited the mutual binding of the two. Overexpression of NAT10 induced PARP1 acetylation followed by increased PARP1 binding to XRCC1, and knockdown of NAT10 expression reduced PARP1 binding to XRCC1. Overexpression of NAT10 enhanced PARP1 binding to LIG3, while knockdown of NAT10 expression decreased PARP1 binding to LIG3. In 10 µmol/L oxaliplatin-treated cells, the γH2AX expression level was 0.38±0.02 in NAT10 overexpressing cells and 1.36±0.15 in NAT10 knockdown cells, both statistically significant (P<0.05) compared with 1.00±0.00 in wild-type cells. In 10 µmol/L oxaliplatin treated cells, the apoptosis rate was (6.54±0.68)% in the NAT10 overexpression group and (12.98±2.54)% in the NAT10 knockdown group, both of which were statistically significant (P<0.05) compared with (9.67±0.37)% in wild-type cells. Conclusion: NAT10 overexpression enhances the binding of NAT10 to PARP1 and promotes the acetylation of PARP1, which in turn prolongs the half-life of PARP1, thus enhancing PARP1 recruitment of DNA damage repair related proteins to the damage sites, promoting DNA damage repair and ultimately the survival of breast cancer cells.

Identification of genes associated with litter size combining genomic approaches in Luzhong mutton sheep.

Litter size is one of the most important reproductive traits of sheep, which has pronounced effects on the profit of husbandry enterprises and enthusiasm of breeders. Despite the importance of litter size, the underlying genetic mechanisms have not been entirely elucidated. Therefore, based on a high-density SNP chip, genome-wide comparative analysis was performed between two groups with different fecundity to reveal candidate genes linked to litter size via detection of homozygosity and selection signatures in Luzhong mutton sheep. Consequently, nine promising genes were identified from six runs of homozygosity islands, and functionally linked to reproduction (ACTL7A, ACTL7B, and ELP1), embryonic development (KLF5 and PIBF1), and cell cycle (DACH1, BORA, DIS3, and MZT1). A total of 128 genes were observed under selection, of which HECW1 and HTR1E were related to total lambs born, GABRG3, LRP1B, and MACROD2 to teat number, and AGBL1 to reproductive seasonality. Additionally, the presence of inbreeding depression implies the urgency of reasonable mating system to increase litter size in the present herd. These findings provide a comprehensive insight to the genetic makeup of litter size, and also contribute to implementation of marker-assisted selection in sheep.

Genome-wide association study of body weight and conformation traits in neonatal sheep.

Sheep, an important source of meat, dairy products and wool, play an essential part in the global agricultural economy. Body weight and body conformation are key traits in the sheep industry; however, their underlying genetic mechanisms are poorly understood. In this study, a GWAS was implemented to identify promising genes possibly linked to birth weight (BW) and body conformation traits in neonatal sheep, using a high-throughput chip (630 K). After quality control, 277 individuals and 518 203 variants were analyzed using gemma software in a mixed linear model. A total of 48 genome-wide suggestive SNPs were obtained, of which four were associated with BW, four with withers height (WH), 11 with body length (BL) and 29 with chest girth (CG). In total, 39 genes associated with BW and body conformation traits were identified by aligning to the sheep genome (Ovis aries_v4.0), and most of them were involved in the cell cycle and body development. Promising candidate genes found included the following: FOS like 2 or AP-1 transcription factor subunit (FOSL2) for BW; potassium voltage-gated channel subfamily D member 2 (KCND2) for WH; transmembrane protein 117 (TMEM117), transforming growth factor beta induced (TGFBI), and leukocyte cell-derived chemotaxin 2 (LECT2) for BL; and trafficking kinesin protein 1 (TRAK1) and LOC101102529 for CG. These results provide cues for similar studies aiming at uncovering the genetic mechanisms underlying body development, and marker-assisted selection programs focusing on BW and body conformation traits in sheep.

Combined approaches to reveal genes associated with litter size in Yunshang black goats.

Intensive artificial selection has been imposed in Yunshang black goats, the first black specialist mutton goat breed in China, with a breeding object of improving reproductive performance, which has contributed to reshaping of the genome including the characterization of SNP, ROH and haplotype. However, variation in reproductive ability exists in the present population. A WGS was implemented in two subpopulations (polytocous group, PG, and monotocous group, MG) with evident differences of litter size. Following the mapping to reference genome, and SNP calling and pruning, three approaches - GWAS, ROH analysis and detection of signatures of selection - were employed to unveil candidate genes responsible for litter size. Consequently, 12 candidate genes containing OSBPL8 with the minimum P-value were uncovered by GWAS. Differences were observed in the pattern of ROH between two subpopulations that shared similar low inbreeding coefficients. Two ROH hotspots and 12 corresponding genes emerged from ROH pool association analysis. Based on the nSL statistic, 15 and 61 promising genes were disclosed under selection for MG and PG respectively. Of them, some promising genes participate in ovarian function (PPP2R5C, CDC25A, ESR1, RPS26 and SERPINBs), seasonal reproduction (DIO3, BTG1 and CRYM) and metabolism (OSBPL8, SLC39A5 and SERPINBs). Our study pinpointed some novel promising genes influencing litter size, provided a comprehensive insight into genetic makeup of litter size and might facilitate selective breeding in goats.

Protease-activated receptor 1 inhibitor improves brain edema in rats with intracerebral hemorrhage.

AIM: To investigate the changes of water content in brain tissue, the expression of AQP4mRNA after cerebral hemorrhage in rats, and the intervention effect of Protease activated receptor 1 inhibitor (PAR1 inhibitor) on both. METHODS: Establish sham operation group (Sham group), ICH group, ICH+PAR1 inhibitor high-dose group (PI(H)group), ICH+PAR1 inhibitor low-dose group (PI(L)group), 25 in each group. Neural dysfunction scores were performed at 1d, 3d, 7d, 14d, and 21d after surgery, and brain water content and AQP4mRNA content were measured. RESULTS: Results: The neurological dysfunction and cerebral edema of rats with cerebral hemorrhage reached the peak at 3 days after operation. With the increase of time, the water content and AQP4mRNA content in the PL(H)group were higher than those in the PI(L)group. The differences were statistically significant. CONCLUSIONS: Appropriate inhibition of PAR1 can alleviate cerebral edema around the hematoma and play a role in improving the function of nerve defects. The mechanism may be realized by down-regulating the expression of AQP4mRNA in brain tissue (Tab. 3, Fig. 3, Ref. 25).

[Research on mental health status of medical staff during COVID-19 epidemic].

Objective: To explore the psychological status of medical staff during COVID-19 epidemic, so as to provide reference and scientific basis for carrying out further psychological intervention and ensuring the mental health of medical staff. Methods: By using convenient sampling method and the Stresss-Anxiety ubscale of Depression Anxiety Stress (DASS-21) , the mental health status of 615 medical staff was investigated by the way of questionnaire star from February 4 to 16, 2020. A total of 615 questionnaires were distributed and collected, and 615 were valid, with an effective recovery rate of 100%. Results: The detection rates of psychological stress and anxiety of medical staff were 13.82% (85/615) and 25.37% (156/615) , respectively. 31-40 years old and working in key departments were risk factors for psychological stress of medical staff (OR=1.779, 2.127) ; Women, frequently washing hands with soap/hand sanitizer/disinfectant were protective factors for psychological stress (OR=0.520, 0.528) . Medical staff working in designated hospitals and key departments were more likely to have anxiety (OR=2.042, 2.702) ; The high fit of the mask to the face and bridge of the nose was a protective factor for the psychological anxiety of medical staff (OR=0.500) . Conclusion: Medical staff show higher stress and anxiety during the epidemic of COVID-19. Psychological intervention should be carried out early, focusing on men, age 31 to 40, medical staff working in designated hospitals and key departments.

[The value of rapid on-site evaluation (ROSE) of bronchoscopy in the diagnosis of pulmonary complications after allogeneic hematopoietic stem cell transplantation].

Objective: To investigate the value of rapid on-site evaluation (ROSE) of bronchoscopy in the diagnosis of pulmonary complications after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods: A retrospective analysis was conducted on the diagnosis and treatment process before and after ROSE examination of 12 patients with pulmonary complications after allo-HSCT who were admitted to the Department of hematology, Tianjin First Central Hospital from January 2017 to June 2019. The diagnostic accuracy of the ROSE was evaluated by comparing the initial diagnosis of ROSE with the final clinical diagnosis. At the same time, the safety of ROSE examination was evaluated and two typical cases were shared. Results: In the 12 transbronchial lung biopsies, there were 5 cases of organizing pneumonia, 3 cases of bronchiolitis obliterans with organizing pneumonia, 1 case of pulmonary fibrosis, 1 case of acute fibrinous and organizing pneumonia, 1 case of pseudomembranous tracheobronchial aspergillosis and 1 case of uncertain diagnosis evaluated by ROSE. Compared with the final clinical diagnosis, there were 10 cases of accurate diagnosis made by ROSE (10/12). All patients were well tolerant to the operation of bronchoscopy. There was only a small amount of bleeding observed during the operation, without pneumothorax and hemoptysis. And no complications related to ROSE occurred. According to the initial diagnosis of ROSE, 10 cases of non-infectious pulmonary complications were treated with methylprednisolone or other immunosuppressive agents and 1 case of Aspergillus infection was given antifungal therapy. Seven patients with non-infectious pulmonary complications improved after treatment. One patient obtained uncertain diagnosis by ROSE was later diagnosed with virus infection by next generation sequencing technology and improved after treatment with foscarnet sodium and immunoglobulin. As of June 30, 2019, 7 patients improved and 5 died. Conclusion: ROSE has the advantages of diagnostic accuracy and rapidity, and is very suitable for patients with critical illness after hematopoietic stem cell transplantation, who are in urgent need of definite diagnosis and prompt treatment, which is beneficial to improve the prognosis of patients.

[HBV genotyping based on key epitopes of PreS1 antigen and its correlation with genotyping by full-length PreS1 sequencing].

Objective: The aim was to investigate the genotype distribution of two major epitopes of large surface protein (PreS1) of hepatitis B in Chinese patients and to explore the association between the genotypes of these two epitopes, and to determine whether PreS1 full-length genotype could be revealed according to the polypeptide sequence of key epitopes. Methods: HBV DNA was extracted from the serum of patients for PCR amplification. 278 samples amplified successfully were sequenced and compared with the known HBV sequences in Genbank to determine the two key epitopes of HBV PreS1 genotype (amino acid epitope 21-47 and 94-117, abbreviated as P21 and P94) and PreS1 full-length genotypes. The correlation among three genotyping approaches was analyzed by Cohen's kappa coefficient to verify the consistency between the key-epitope genotyping and the full-length preS1 genotyping. Results: 232 samples were successfully sequenced. The genotyping based on the kind of P21 epitope protein sequence, 201 cases for genotype C, 23 cases for genotype B and 8 cases for uncertain genotypes and genotyping based on the form of P94 epitope protein sequence, 199 cases for genotype C, 25 cases for genotype B and 8 cases for indeterminate genotypes. Lastly, the genotyping based on sequence of the full-length PreS1 sequence, 207 and 25 cases for genotype C and B. P21 or P94 epitope genotyping and PreS1 full length genotyping were highly consistent, respectively, 96.55% and 96.12%, and the two epitopes (P21and P94) genotyping have parallel consistency (93.10%). Conclusion: In this study, an innovatively genotyping method based on the amino acid sequence of key epitopes was proposed. The genotypes of HBV in china were mainly B and C genotypes, and the genotypes of key conserved epitopes of HBV PreS1 were highly consistent with the full-length genotyping ( > 96%). Moreover, genotyping with one or two key epitopes can be used in place of the full-length genotyping.

[Research of mHLA-G and the receptor expression with kidney rejection and CMV active infection following kidney transplantation].

Objective: To study the expression of membrane HLA-G (mHLA-G) and the receptor immunoglobulin-like transcript 2(ILT2) on lymphocyte and find their association with rejection and cytomegalovirus (CMV) infection after renal transplantation. Methods: A total of 88 cases of renal transplant recipients for the first time from February 2014 to February 2016 were studied in this work. Recipients can be divided into rejection group (n=12) and stable renal function group (n=41) according to whether rejection occurred. Recipients only infected CMV not developed rejection were included in the CMV positive group (n=24). CMV negative group (n=11) including CMV negative recipients once infected CMV.The expression of mHLA-G and ILT2 on lymphocytes were detected by flow cytometry, and the differences among different groups were analyzed. Results: The data showed that after renal transplantation, T and B lymphocytes mHLA-G expression rate was the lowest in the rejection group (0.42%±0.35%, 0.88%±0.47%), having significant difference with renal function stable group and CMV positive group (all P<0.01). In CMV positive group the expression of mHLA-G on T and B lymphocytes was the highest (1.31%±0.69%, 2.01%±0.91%), having significant difference with rejection group (P<0.001). The expression of mHLA-G on B cell was statistically significantly different between CMV positive group and CMV negative group (P<0.05). There was no significant difference in ILT2 expression on B cell among the four groups (P>0.05). The expression rate of ILT2 on T cells was higher in the CMV positive group (36.91%±14.91%), having significant difference with the other three groups (P<0.01). Conclusions: Low expression of mHLA-G on T and B lymphocytes may predict rejection after renal transplantation. High expression of mHLA-G and ILT2 on lymphocytes is prone to CMV infection after renal transplantation .

[Impact of improvement of diabetes on survival after radical pancreatic resection in patients with pancreatic cancer and diabetes].

OBJECTIVE: To explorer the effect of improvement of diabetes on survival after radical pancreatic resection in patients with pancreatic cancer and diabetes. METHODS: Seventy-three patients who underwent radical resection for pancreatic cancer at Ruijin Hospital (Shanghai, China) between July 2007 and December 2011 and had preoperative diabetes were retrospectively analyzed. Clinical characteristics and overall postoperative survival were compared between patients with controlled/uncontrolled diabetes. Among patients with uncontrolled diabetes, comparison was made in clinical characteristics and overall postoperative survival between patients with long-term (≥2 years) diabetes and those with new-onset (<2 years) diabetes. RESULTS: Controlled diabetes after radical pancreatic resection was observed in 38.4% (28/73) in the patients with pancreatic cancer and diabetes. The median follow-up time was 44.5 months (95CI: 31.7-57.2 months). Patients in whom diabetes was controlled had well-differentiated tumors and longer overall survival compared to patients with uncontrolled diabetes [median overall survival (95%CI) 41.0 (34.2-47.7) months vs 17.0 (14.8-19.2) months, P<0.001]. Among the 45 patients with uncontrolled diabetes, there was no differences in clinic characteristic and overall survival between new-onset (n=21) and long-term diabetes (n=24) [median overall survival (95%CI): 17.8 (15.7-20.1) months vs 15.8 (10.5-21.1) months, P=0.198]. CONCLUSION: Good postoperative control of diabetes after radical pancreatic resection in patients with pancreatic cancer and diabetes may predict favorable survival.

[Expression and significance of neutrophil surface adhesion molecules in renal transplant recipients with cytomegalovirus infection].

OBJECTIVE: To study the expression and its diagnostic significance of neutrophil surface adhesion molecules including CD11b, CD15 and CD62L after renal transplantation in recipients with cytomegalovirus (CMV) infection. METHODS: Blood samples were collected from 142 kidney transplant recipients, including 95 males and 47 females, who received allogeneic renal transplantation between September 2009 and January 2015 in 309th Hospital of the PLA. Healthy volunteers (22 males and 9 females) were recruited from physical examination center in 309th Hospital of the PLA from September 2009 to January 2015 as healthy control group. Renal transplant recipients were divided into high active CMV infection group, active CMV infection group and CMV negative control group according to CMV-pp65 antigen detection. Neutrophil surface adhesion molecules CD11b, CD15 and CD62L were detected by flow cytometry and their mean fluorescence intensity compared among the groups. Receiver operating characteristic (ROC) curves of CD11b, CD15 and CD62L in detecting active infection in renal transplant recipients were made. RESULTS: The mean fluorescence intensity of CD15 in high active CMV infection group(n=17) and active CMV infection group(n=65)were 776.31±89.53 and 554.39±67.89, respectively, with significant differences compared with CMV negative control group (n=60, 334.92±44.69) and healthy control group (n=31, 310.56±39.67) (all P<0.05); the expression proportions of CD11b and CD62L in high active CMV infection group and were 42.31%±6.11% and 40.35%±6.47%, respectively, with significant differences compared with active CMV infection group(62.45%±5.67% and 65.65%±5.33%), CMV negative control group(70.74%±6.55% and 70.37%±6.71%) and healthy control group(72.52%±6.48% and 72.43%±6.51%) (all P<0.05). The optimal cut-off values of CD11b and CD62L in diagnosing active CMV infection group were 56.61% and 44.35%, respectively, with the sensitivity being both 100.00%, the specificity being 76.67% and 58.06% respectively, and the area under the curve (AUC) being 0.851 and 0.628 respectively; the optimal cut-off values of CD11b and CD62L in diagnosing high active CMV infection group were 66.57% and 69.56% respectively, with the sensitivity being 81.54% and 87.69% respectively, the specificity being 100.00% and 98.33% respectively, and the AUC being 1.000 and 0.991 respectively; the optimal cut-off values of mean fluorescence intensity of CD15 in diagnosing high active CMV infection group and active CMV infection group were 542.71 and 408.03 respectively, the sensitivity in the two groups being 100.00% and 98.46% respectively, the specificity being both 100.00%, and the AUC being 1.000 and 0.999 respectively. CONCLUSIONS: Neutrophils CD15 expression may be up-regulated in renal transplantation recipients with CMV infection, while neutrophils CD11b and CD62L expressions are down-regulated. Such changes in CD15, CD11b and CD62L expression can be used as a basis for laboratory diagnosis of active CMV infection.

[Establishment of macrophage model of iron overload in vitro and the injury induced by oxidative stress on macrophage with iron overload].

OBJECTIVE: To establish macrophage iron overload model in vitro by co-culture macrophages with iron, and to explore the effect of iron overload on cell reactive oxygen species (ROS) and the impact of ROS on macrophages. METHOD: Iron overload group were treated with different concentrations (0, 5, 10, 20, 40, 80 µmol/L respectively) of ferric ammonium citrate (FAC). The control group was the group of macrophages without FAC treatment. We detected the number and state of cells, metabolic activity, the change of phagocytosis, the levels of ROS and reactive nitrogen, and changes of related oxidative stress signaling pathways in different groups. Changes in the above indexes were detected after application of deferasirox (DFX) to remove iron and the antioxidant N -acetylcysteine (NAC) to clear excess oxidative stress. RESULTS: (1)The levels of labile iron pool (LIP) in macrophages co-cultivated with iron was increased with the increase of iron concentration in a dose-dependent manner. The LIP levels was the highest in the macrophages treated with 80 µmol/L. (2)The increase of FAC concentration, the metabolic activity of macrophages in the 5 FAC-treated groups decreased to 51.58%, 40.98%, 16.23%, 3.46%, and 0.05% of the activity level of the control group (all P< 0.05). The group with the metabolic activity decreased to 16.23% (20 µmol/L) was selected as the iron overload group for the following experiments. (3)Compared with the control group, the number of macrophages in the iron overload group reduced to 32.80% (P<0.05), and the state of cells changed from adherence to partial suspension. The phagocytosis of macrophages in the iron overload group reduced to 20.40% of the control group (P<0.05). (4)Our further experiment showed that the levels of ROS and the activity nitrogen in the iron overload group increased by 7.71-and 1.45-fold compared with the control group (both P<0.05). The RT-PCR showed up-regulated mRNA expression of genes related with ROS production, i. e. nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX 4) gene related with ROS production and inducible nitric oxide synthase (iNOS) gene related with reactive nitrogen production, down-regulated mRNA expression of glutathione peroxidase 1 (GPX1) gene which participated in ROS clearance. Moreover, mRNA expression of phosphatidylinositol-3-kinase (PI3K) gene involved in oxidative stress signaling pathway in the iron overload group was up-regulated, while fork head protein O3 (FOXO3) which regulated oxidative stress through negative feedback showed a down-regulation level of mRNA expression compared with the control group. (5)After iron chelation and antioxidant treatment, the above-mentioned damage in the iron overload group were partially reversed. CONCLUSIONS: The damages of iron overload on macrophages may be mediated by inducing oxidative stress and activating oxidative stress signaling pathways. Our established model provides a method to explore the mechanism of iron overload on macrophage, and may shed some new light on possible therapeutic target in treating iron overload patients.

[Value of MRI in prenatal diagnosis of abnormal fetal kidneys].

Objective: To investigate the value of MRI in the prenatal diagnosis of abnormal fetal kidneys. Methods: From December 2014 to March 2016, 51 women underwent MRI and were confirmed as having fetuses with abnormal fetal kidneys when follow up. Their clinical and MRI profiles were analyzed retrospectively, including MRI manifestation, the fetal kidney signal intensity of diffusion-weighted imaging (DWI) and apparent diffusion coefficient (ADC). The signal intensity of DWI and ADC of the abnormal kidney and the normal opposite kidney, and those of the normal and abnormal kidneys in different individuals were compared. The accuracies of MRI and ultrasound in evaluating abnormal fetal kidneys were also compared. Results: MRI could accurately demonstrate the morphological features of abnormal fetal kidneys. There was no statistically significant difference between DWI signal intensity and ADC value of the abnormal kidney[368 ± 125, (1.516 ± 0.420) × 10-3 mm2/s] and the normal opposite one[410 ± 125, (1.362 ± 0.251) × 10-3 mm2/s], P values were 0.165 and 0.184, respectively. The DWI signal intensity of normal kidneys (401 ± 124) was higher than that of renal cysts (182 ± 40, P <0.01), and the ADC value of normal kidneys[(1.306 ± 0.252) × 10-3 mm2/s] was lower than that of renal cysts[(2.912 ± 0.235) × 10-3 mm2/s] and multicystic dysplastic kidneys[(1.870±0.654) ×10-3 mm2/s], P values were <0.01 and 0.045, respectively. The diagnostic accuracy of MRI and prenatal ultrasound for abnormal fetal kidneys were 94% (63/67) and 85% (57/67), respectively. However, there was no statistical difference between the two methods (P=0.070). Conclusion: MRI have an important role in the prenatal diagnosis and evaluation of abnormal fetal kidneys.