[Opportunities and challenges in accurate early diagnosis and treatment of lung cancer/pulmonary nodules].

Lung cancer remains the most prevalent and lethal malignancy in our country. Early diagnosis and treatment are crucial for improving patient prognosis in lung cancer/pulmonary nodules. Recent advancements in non-invasive/minimally invasive liquid biopsy, multi-omics, and artificial intelligence technologies have significantly enhanced the accuracy of early lung cancer/pulmonary nodule diagnosis. However, an early diagnostic method with both high sensitivity and specificity is yet to be established. Furthermore, addressing the methods and extent of early precision surgery, local precision therapy, perioperative combined treatment, and postoperative recurrence and metastasis monitoring are urgent challenges in the early management of lung cancer/pulmonary nodules. Integrating the advantages of various treatment strategies and formulating personalized and precise treatment plans is key to further improving patient survival. In the future, while exploring new therapeutic strategies, it is necessary to continuously search for biomarkers to identify the population that will benefit from the treatment effectively. Additionally, large-sample randomized controlled clinical studies should be conducted to investigate the benefits of long-term patient survival under a diverse range of treatment strategies.

Single-cell and bulk RNA sequencing reveal heterogeneity and diagnostic markers in papillary thyroid carcinoma lymph-node metastasis.

PURPOSE: Papillary thyroid carcinoma (PTC) is characterized by lymph-node metastasis (LNM), which affects recurrence and prognosis. This study analyzed PTC LNM by single-cell RNA sequencing (scRNA-seq) data and bulk RNA sequencing (RNA-seq) to find diagnostic markers and therapeutic targets. METHODS: ScRNA-seq data were clustered and malignant cells were identified. Differentially expressed genes (DEGs) were identified in malignant cells of scRNA-seq and bulk RNA-seq, respectively. PTC LNM diagnostic model was constructed based on intersecting DEGs using glmnet package. Next, PTC samples from 66 patients were used to validate the two most significant genes in the diagnostic model, S100A2 and type 2 deiodinase (DIO2) by quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and immunohistochemical (IHC). Further, the inhibitory effect of DIO2 on PTC cells was verified by cell biology behavior, western blot, cell cycle analysis, 5-ethynyl-2'-deoxyuridine (EdU) assay, and xenograft tumors. RESULTS: Heterogeneity of PTC LNM was demonstrated by Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analysis. A total of 19 differential genes were used to construct the diagnostic model. S100A2 and DIO2 differ significantly at the RNA (p < 0.01) and protein level in LNM patient tissues (p < 0.001). And differed in PTC tissues with different pathologic typing (p < 0.001). Further, EdU (p < 0.001) and cell biology behavior revealed that PTC cells overexpressed DIO2 had reduced proliferative capacity. Cell cycle proteins were reduced and cells are more likely to be stuck in G2/M phase (p < 0.001). CONCLUSIONS: This study explored the heterogeneity of PTC LNM using scRNA-seq. By combining with bulk RNA-seq data, diagnostic markers were explored and the model was established. Clinical diagnostic efficacy of S100A2 and DIO2 was validated and the treatment potential of DIO2 was discovered.

[Pathological significance of plasma cell infiltration in diagnosing lymph node diseases].

Objective: To investigate the value of plasma cells for diagnosing lymph node diseases. Methods: Common lymphadenopathy (except plasma cell neoplasms) diagnosed from September 2012 to August 2022 were selected from the pathological records of Changhai Hospital, Shanghai, China. Morphological and immunohistochemical features were analyzed to examine the infiltration pattern, clonality, and IgG and IgG4 expression of plasma cells in these lymphadenopathies, and to summarize the differential diagnoses of plasma cell infiltration in common lymphadenopathies. Results: A total of 236 cases of lymphadenopathies with various degrees of plasma cell infiltration were included in the study. There were 58 cases of Castleman's disease, 55 cases of IgG4-related lymphadenopathy, 14 cases of syphilitic lymphadenitis, 2 cases of rheumatoid lymphadenitis, 18 cases of Rosai-Dorfman disease, 23 cases of Kimura's disease, 13 cases of dermal lymphadenitis and 53 cases of angioimmunoblastic T-cell lymphoma (AITL). The main features of these lymphadenopathies were lymph node enlargement with various degrees of plasm cell infiltration. A panel of immunohistochemical antibodies were used to examine the distribution of plasma cells and the expression of IgG and IgG4. The presence of lymph node architecture could help determine benign and malignant lesions. The preliminary classification of these lymphadenopathies was based on the infiltration features of plasma cells. The evaluation of IgG and IgG4 as a routine means could exclude the lymph nodes involvement of IgG4-related dieases (IgG4-RD), and whether it was accompanied by autoimmune diseases or multiple-organ diseases, which were of critical evidence for the differential diagnosis. For common lesions of lymphadenopathies, such as Castleman's disease, Kimura's disease, Rosai-Dorfman's disease and dermal lymphadenitis, the expression ratio of IgG4/IgG (>40%) as detected using immunhistochemistry and serum IgG4 levels should be considered as a standard for the possibility of IgG4-RD. The differential diagnosis of multicentric Castleman's diseases and IgG4-RD should be also considered. Conclusions: Infiltration of plasma cells and IgG4-positive plasma cells may be detected in some types of lymphadenopathies and lymphomas in clinicopathological daily practice, but not all of them are related to IgG4-RD. It should be emphasized that the characteristics of plasma cell infiltration and the ratio of IgG4/IgG (>40%) should be considered for further differential diagnosis and avoiding misclassification of lymphadenopathies.

[A multicenter clinical trial of collar-button type keratoprosthesis implantation for the treatment of corneal blindness in high-risk transplantation].

Objective: To evaluate the efficiency and safety of collar-button type keratoprosthesis (c-bKPro) implantation for corneal blindness in high-risk transplantation in China. Methods: It was a case series study. High-risk corneal blind patients who planned to undergo c-bKPro implantation were prospectively and continuously enrolled in the Eye Hospital of Shandong First Medical University, Ophthalmology Division of Chinese PLA General Hospital, Zhongshan Ophthalmic Center, Department of Ophthalmology in Eye & ENT Hospital of Fudan University, and Eye Hospital of Wenzhou Medical University from July 2019 to January 2020. The cure for blindness and surgical success were assessed based on visual acuity (VA)≥0.05. The complications and keratoprosthesis retention rate were recorded to determine the safety of the surgery. Results: Thirty-seven subjects (eyes) were included, of which 32 were male and 5 were female, aged 27 to 72 years old. The indications of c-bKPro implantation were corneal graft failure (21 eyes, 56.8%), chemical injury (8 eyes, 21.6%), thermal burn (5 eyes, 13.5%), unexplained corneal opacity (2 eyes, 5.4%), and corneal perforation (1 eye, 2.7%). Two patients withdrew from the clinical trial at 3 months postoperatively. Thirty-five patients were followed up for 6 months, and 31 were followed up for 12 months. The VA was ≥0.05 in 83.8% of eyes at 6 months and in 81.8% of eyes at 12 months. Among the 11 eyes diagnosed with concurrent glaucoma, 6 eyes achieved a VA of ≥0.05. At 12 months, the c-bKPro retention rate was 100%. The surgical complications included retroprosthetic membrane formation (5 eyes, 16.1%), persistent corneal epithelial defects (5 eyes, 16.1%), macular edema (4 eyes, 12.9%), new-onset glaucoma (4 eyes, 12.5%; including one eye withdrawn from the study at 3 months), sterile corneal melting (2 eyes, 6.5%), sterile vitritis (1 eye, 3.2%), and infectious keratitis (1 eye, 3.2%). Conclusions: C-bKPro implantation is an effective and safe option for treating corneal blindness in high-risk transplantation in China. Improved visual outcomes could be achieved in most cases, with a relatively low incidence of postoperative complications.

Whole-lesion histogram analysis of mono-exponential and bi-exponential diffusion-weighted imaging in differentiating lung cancer from benign pulmonary lesions using 3 T MRI.

AIM: To investigate whether whole-lesion histogram analysis of apparent diffusion coefficient (ADC) values derived from mono-exponential and bi-exponential diffusion-weighted imaging (DWI) can differentiate lung cancer from benign pulmonary lesions. MATERIALS AND METHODS: Thirty-two patients with lung cancer and 17 patients with benign pulmonary lesions were included retrospectively. All patients underwent DWI before surgery or biopsy. ADC histogram parameters, including mean, percentile values (10th and 90th), kurtosis, and skewness, were calculated independently by two radiologists. The histogram parameters were compared between patients with lung cancer and benign lesions. Receiver operating characteristic curves were constructed to evaluate the diagnostic performance. RESULTS: The ADCMean, ADC10th, DMean, D10th were significantly lower in lung cancer (1.187 ± 0.144 × 10-3; 0.440 ± 0.062 × 10-3; 1.068 ± 0.108 × 10-3; and 0.422 ± 0.049 × 10-3 mm/s) compared to benign lesions (1.418 ± 0.274 × 10-3; 0.555 ± 0.113 × 10-3; 1.216 ± 0.149 × 10-3; and 0.490 ± 0.044 × 10-3 mm/s; p<0.05). The ADCSkewness and DSkewness were significantly different between lung cancer (2.35 ± 0.72; 2.58 ± 1.14) and benign lesions (1.85 ± 0.54; 1.59 ± 1.47; p<0.05). D10th was robust in differentiating lung cancer from benign lesions. Using 0.453 × 10-3 mm/s as the optimal threshold, the sensitivity, specificity, and accuracy of D10th were 78.12%, 82.35%, and 79.6%, respectively. CONCLUSION: Whole-lesion histogram analysis of ADC values derived by mono-exponential and bi-exponential DWI using 3 T magnetic resonance imaging helps distinguish lung cancer from benign pulmonary lesions.

[Cytological analysis of lymphoblastic lymphoma/acute lymphoblastic leukemia in serous effusion].

Objective: To study the diagnostic clues and significance in serous effusion cytology associated with lymphoblatic lymphoma/acute lymphoblastic leukemia (LBL/ALL). Methods: Forty-five serous effusion specimens with final diagnosis of LBL/ALL were collected from August 2011 to December 2019 at the First Affiliated Hospital of Zhengzhou University. All cases were reviewed for their clinical profiles, cytomorphologic features and ancillary studies. Cell blocks and immunocytochemistry were prepared in 22 cases; flow cytometric immunophenotyping was performed in three cases and gene rearrangement analysis (T-cell recepter, TCR and immunoglobulin, Ig) was performed in five cases. Results: Among the 45 cases, there were 35 males and 10 females with male to female ratio of 3.5∶1.0. The median age was 15 years. Mediastinal mass was the initial presentation in 39 patients (86.7%) and high LDL level were observed in 34 patients (75.6%). Microscopically, the majority of the specimens (86.7%) were hypercellular. The smears demonstrated dispersed lymphoblasts that were predominantly small to intermediate in size with scanty basophilic cytoplasm and irregular or convoluted nuclei with fine chromatin condensation and inconspicuous nucleoli. Mitoses were frequently observed. Karyorrhexis and apoptosis were seen in all cases. By immunophenotyping, TdT was expressed in 19 cases (86.4%) and CD99 in 20 cases (90.9%). Ki-67 expression varied from 65% to 95%. Flow cytometry in three cases demonstrated positivity for TdT, CD2, CD3 and CD7. Monoclonal TCR gene rearrangement was found in 4 of 5 cases, and both monoclonal TCR and Igκ gene were found in 1 case. Conclusions: In LBL/ALL, primary diagnosis could be made basing on clinical features (younger male patients with a mediastinum mass) and cytomorphology (monotonous, small to medium sized lymphoid cells with prominent irregular nuclei, fine chromatin and frequent mitoses, karyorrhexis and apoptosis). If immunocytochemistry and other ancillary studies are performed, the accuracy and reliability of the results could be improved.

[Metastatic renal cell carcinoma: a clinicopathological analysis of 196 cases].

Objective: To analyze the clinico pathological features, differential diagnosis and prognosis of metastatic renal cell carcinomas. Methods: The clinical data, histology, immunophenotype and follow-up data of 196 patients with metastatic renal cell carcinoma diagnosed from 1994 to 2017 at the Department of Pathology, Changhai Hospital, Naval Military Medical University, Shanghai, China were analyzed retrospectively. Results: There were 142 males and 54 females, with a median age of 61 years. The top three metastatic sites for the 196 cases of metastatic renal cell carcinoma were lung (31.1%, 61/196), bone (29.1%, 57/196) and digestive system (19.4%, 38/196). Among the pathological subtypes of metastasis, the proportion of clear cell renal cell carcinoma was 94.4% (185/196) and that of type II papillary renal cell carcinoma was 3.6% (7/196). The TFE3 translocated renal cell carcinoma and congestive tubular carcinoma were rare, with 3 cases and 1 case, respectively. CK, vimentin, CAⅨ and CD10 were expressed in all metastatic clear cell renal cell carcinomas. CK7, CD10 and P504s were expressed in papillary renal cell carcinomas. TFE3 was expressed in TFE3 translocated renal cell carcinoma. The collecting duct carcinoma was positive for HCK. Conclusions: Lung metastasis and bone metastasis are still the most frequent metastatic sites of renal cell carcinoma. Five years after primary lesion resection may be the high risk time for metastasis. Most of the metastases are solitary when they are first identified. To better diagnose and identify the renal origin of a metastatic renal cell carcinoma, one should consider morphological characteristics, clinical history information of the metastasis and the combined immunohistochemistry of CK, vimentin, CD10, CK7, TFE3, PAX2 and PAX8.

[Clinical application of the gasless unilateral axillary approach in endoscopic thyroid surgery].

Objective: To investigate the efficacy, safety and advantages of gasless unilateral axillary approach (GUAA) in endoscopic thyroid surgery. Methods: A total of 334 patients who underwent the GUAA endoscopic thyroid surgery (GUAA group) or conventional open thyroid surgery (OS group) in the Department of Head and Neck Surgery of Zhejiang Cancer Hospital from January 2017 to June 2018 were retrospectively analyzed. There were 45 males and 289 females, aged from 12 to 72 years old, of whom 139 patients were assigned to GUAA group and 195 patients to OS group. Pathological results included papillary thyroid carcinoma (282 cases), nodular goiter (41 cases) and thyroid adenoma (11 cases). Surgical exploration development curve of GUAA group was drawn and was divided into two parts: the technical exploration stage and the technical stable stage. Surgical efficiency, incidences of complications, and incision satisfaction were compared between GUAA group in technical stable stage and OS group. SPSS 25.0 software was adopted for statistical analysis. Results: The mean age in GUAA group was younger than that in OS group, with a significant difference [(35.3±9.5) years vs. (48.1±10.6) years, t=11.31, P<0.01]. The cases in the endoscope group were divided into technical exploration stage for 51 cases and technical stable stage for 88 cases according to the exploration and development curve. In unilateral radical thyroidectomy and unilateral thyroid lobectomy, the mean operation time [(90.6±18.6) min and (93.5±22.0) min] and postoperative drainage volumes [(121.5±87.6) ml and (155.5±69.1) ml] of GUAA group in the stable stage were more than those of OS group [(61.6±15.6) min and (46.5±8.4) min] and [(93.2±42.3) ml and (78.9±48.7) ml]. The difference was statistically significant (t=12.28, 7.23, 3.35 and 3.05 respectively, all P<0.05), but there were no significant differences in surgical bleeding volumes between two groups [(12.7±6.8) ml vs. (13.5±7.7) ml, t=0.74, P>0.05 and (16.3±14.1) ml vs. (11.9±5.1) ml, t=1.05, P>0.05]. Compared with OS group, GUAA group had the lower incidence of anterior cervical discomfort during swallowing (2.3% vs. 29.2%, P<0.01) and the higher incision satisfaction score (1.1±0.5 vs. 2.8±0.7, t=21.12, P<0.01), however, GUAA group had the higher incidence of supraclavicular (or infraclavicular) numbness after surgery (5.7% vs. 0, P<0.01). And there was no significant difference in the incidences of temporary recurrent laryngeal nerve injury, bleeding, hematoma, infection, lymphatic leakage or chylous leakage after surgery between two groups (P>0.05). Conclusion: GUAA endoscopic thyroid surgery is a safe method with high cosmetic satisfaction.

Gelatin methacryloyl hydrogel eye pad loaded with amniotic extract prevents symblepharon in rabbit eyes.

OBJECTIVE: The aim was to evaluate the ability of gelatin methacryloyl (GelMA) hydrogel eye pads loaded with amniotic extract to prevent symblepharon in rabbits. MATERIALS AND METHODS: Forty-eight rabbits were divided into 3 groups. After ocular alkali burn, Group A (n=16) was treated with amniotic extract-loaded hydrogel eye pads placed in the conjunctival sac, Group B (n=16) was treated with amniotic membrane transplantation, and Group C (n=16) received no treatment. At 1, 2, 3, and 4 weeks post-injury, 4 rabbits from each group were selected to evaluate for symblepharon, determine epithelial healing rate and corneal neovascularization, conduct histopathology, and to quantify the expression of TGF-ß1. RESULTS: At 1 week post-injury, the epithelial healing rate in Groups A and B was higher than Group C (p=0.002, 0.001, respectively). At 2 weeks, corneal neovascularization in Group B was less than Group C (p=0.004). At 3 and 4 weeks, no symblepharon has been found in Group A, but it was found in some eyes in Group B and C (p=0.009, 0.013). Further, the expression of TGF-ß1 in Group A was lower than in Group B and C (p<0.001). H&E staining showed that the controls in Group C had more edema and inflammatory cell infiltration in the first 2 weeks, relative to Groups A and B. At 4 weeks, Masson's Trichrome staining showed that fibers were most regularly aligned in Group A and that immuno-histochemical staining found that proliferating cell nuclear antigen was highest expressed in Group C. CONCLUSIONS: Treatment with GelMA hydrogel eye pads loaded with amniotic extract shortly after chemical injury prevented symblepharon in rabbits.

Curcumin exerts protective effect on PC12 cells against lidocaine-induced cytotoxicity by suppressing the formation of NLRP3 inflammasome.

OBJECTIVE: To explore the protective effect of curcumin on lidocaine-insulted PC12 cells. MATERIALS AND METHODS: We first treated PC12 cells with different doses of lidocaine, and then treated the cells with curcumin or Nod-like receptor pyrin domain3 (NLRP3) inhibitor (MCC950). Subsequently, the cell viability, apoptosis, reactive oxygen species (ROS) production and NLRP3 inflammasome were detected by cell counting kit-8 (CCK8), Annexin V/PI staining, FCM and Western blot analysis, respectively, and the level of IL-1ß in PC12 cells was determined by an enzyme-linked immunosorbent assay (ELISA) kit. RESULTS: Lidocaine inhibited the viability of PC12 cells, and it induced cell apoptosis, promoted ROS release and activated NLRP3 inflammasome in PC12 cells, but its effects were reversed by the treatment of curcumin. Moreover, NLRP3 over-expression also induced cytotoxicity in PC12 cells, which was also rescued by the treatment of curcumin. CONCLUSIONS: Our study indicates that curcumin exerts protective effect against lidocaine-induced cytotoxicity on PC12 cells by suppressing the activity of NLRP3 inflammasome, which provides new ideas on screening natural product for neurological damage therapy.

[Long-term prognostic value of mean platelet volume in patients with stable coronary artery disease undergoing elective percutaneous coronary intervention].

Objective: To evaluate the relationship between admission mean platelet volume (MPV) and 2-year cardiac mortality in patients with stable coronary artery disease (CAD) undergoing elective percutaneous coronary intervention (PCI), and explored the consistence of this relationship in diabetes mellitus (DM) and non-DM subgroups. Method: A total of 4 293 patients who underwent PCI in Fuwai Hospital in 2013 were enrolled and divided into two groups according to MPV as follows: lower MPV (n=2 219, MPV≤10.5fL) and higher MPV (n=2 074, MPV>10.5fL). Result: Patients with high MPV had a higher rate of DM (30.4%(674/2 219) vs 34.5%(715/2 074)), smoking (53.3%(1 183/2219) vs 57.0%(1 182/2 074)), and previous coronary artery bypass grafting (CABG) (4.0%(88/2 219) vs 5.4%(112/2 074)), while left ventricular ejection fraction (LVEF) (64±7 vs 63±7), and glomerular filtration rate (eGFR) (92±14 vs 91±15) were lower compared with patients in the low MPV group (all P<0.05). In the laboratory examination, patients with high MPV had higher glycosylated hemoglobin, and lower platelet count (all P<0.05). In coronary angiography, there was no significant difference in SYNTAX scores, left main/three-vessel lesions, stent type, success rate of operation, and total stent length (all P>0.05). Compared with low MPV group, patients with high MPV had ahigher cardiac mortality [18 (0.9%) vs 5 (0.2%), P=0.004]. Kaplan-Meier analysis showed that compared to low MPV group, cardiac mortality in high MPV group was significantly higher (Log-rank P=0.004). Multivariate Cox regression analysis showed that high MPV was independently associated with 2-year cardiac mortality (HR 4.127, 95%CI 1.373 to 12.405, P=0.012). Receiver operating characteristic curve (ROC) analysis also showed that MPV had a good diagnostic value in predicting 2-year cardiac mortality (area under the curve=0.624, 95%CI: 0.511-0.738, P=0.04). Subgroup analysis showed that in patients with DM (HR 2.090, 95%CI 1.217-3.589, P=0.008) and male (HR 1.561, 95%CI 1.007-2.421, P=0.047), MPV was significantly related with cardiac mortality. Conclusion: In patients with stable CAD who underwent elective PCI, high MPV was independently associated with an increase in 2-year cardiac mortality, especially in patients with DM and male gender.

[Split-thickness scalp and allogenic acellular dermal matrix in repairing deep wounds of hands in patients with extremely extensive burns].

Objective: To explore the clinical effects of split-thickness scalp and allogenic acellular dermal matrix (ADM) in repairing deep wounds of hands in patients with extremely extensive burns. Methods: Six patients with extremely extensive burns complicated by deep burn in hands were admitted to our department from December 2014 to December 2017, including 4 males and 2 females, aged 21 to 58 years. Their total burn areas were from 85% to 95% total body surface area (TBSA), and the sum of deep-partial thickness and full-thickness wounds was larger than 50% TBSA. Under general anesthesia, deep wounds of 10 dorsal hands were repaired by combined transplantation of split-thickness scalp and allogenic ADM in 4 to 6 weeks after injury. The skin grafting range was beyond the metacarpophalangeal joint, including partial dorsal digital deep wounds. Survival of skin grafts on dorsal hands, follow-up of hand shape and functional recovery were observed and recorded. Results: Six patients were successfully treated. There was no infection on the dorsal deep wounds of 10 hands after surgery, and the grafts survived well. During follow-up of 1 to 2 years after operation, there were no recurrent tension blisters or dorsal extension deformity of the metacarpophalangeal joints in 10 hands, the shape and function of hands recovered well, and hand Carroll scores were from 90 to 99 points, with functional levels of Ⅴ and Ⅵ. Conclusions: The combined transplantation of split-thickness scalp and allogenic ADM is a good method for repairing deep hand wounds in patients with extremely extensive burns, which can alleviate the cicatrix hyperplasia and contracture of healed hand wounds, and improve the shape and function of hands.

[The role and mechanism of TRAF2 in the biological behavior of gastric cancer].

Objective: To clarify the effect of TRAF2 in the biological behavior of gastric cancer and explore the mechanism. Methods: TRAF2 stably depleted AGS cell was established. Cell growth was monitored by x-CELLigence system. Cell proliferation was detected using cell viability assay. The apoptosis and cell cycle were detected by flow cytometry. The difference of migration and invasion abilities were measured by real-time xCELLigence system and Transwell. The expression and activity of NF-κB signaling pathway were measured by western blot and TransAM assay. The expression of TRAF2 in gastric cancer tissue and its clinical significance were detected by immunohistochemistry. Results: The cell index of AGS-siTRAF2 cells was significantly lower than that of AGS-sictrl cells at 8 h. In the cell viability assay, the A values of AGS-siTRAF2 cells were 51 296.00±2 631.06, 68 389.25±6 703.21 and 65 559.50±6 339.22 at 24 h, 48 h and 72 h. The values of the viability of AGS-siTRAF2 cells were significantly lower than those of AGS-sictrl cells (P<0.001). The results of flow cytometry showed that the apoptosis rates of AGS-siTRAF2 cells were (1.42±0.07)%, (2.98±0.11)% and (1.56±0.03)% at 24 h, 48 h and 72 h, respectively, which were significantly higher than those of AGS-sictrl cells (all P<0.05). The distribution of S phase in AGS-siTRAF2 cells was (23.57±1.12)%, while that in the AGS-sictrl cells was (19.49±1.19)%. The difference was statistically significant (P=0.012). AGS-siTRAF2 cells migrated much slower than AGS-sictrl cells from 3 h and the number of migrated AGS-sictrl cells was 121.7±6.7 while that of AGS-siTRAF2 cells was 84.0±6.6 (P=0.002). The cell index of AGS-siTRAF2 cells was less than that of AGS-sictrl cells from 3 h. In Transwell assay, the number of invaded AGS-sictrl cells was 109.3±3.1 after 24 h of culture, significantly higher than 79.0±6.2 of AGS-siTRAF2 cells (P=0.002). Western blot analysis showed that the expression levels of RelA, RelB, p50 and p52 in AGS-siTRAF2 cells were significantly lower than those in AGS-sictrl cells. The activities of RelA, RelB, p50 and p52 in AGS-siTRAF2 cells were 0.01±0.00, 0.01±0.01, 0.92±0.01 and 0.53±0.03, respectively, significantly lower than those of AGS-sictrl cells (all P<0.001). High TRAF2 expression (TRAF2-high) was found in 53.0% of GC samples, while TRAF2-high was only observed in 38.0% of the paired adjacent tissues (P=0.033). The expression of TRAF2 was significantly higher in the tubular adenocarcinoma, poor differentiation advanced T, advanced N, and clinical staging (P<0.05). The median survival time were 17 months and 78 months in the TRAF2 high-expression and low-expression groups, respectively, and the difference was statistically significant (P=0.010). Conclusions: Depletion of TRAF2 inhibits the AGS cell growth, migration and invasion. The expression of TRAF2 is increased in gastric tumor tissue. The expression of TRAF2 is associated with the prognosis of gastric cancer.

[Clinical characteristics and prognosis between male and female patients with premature coronary artery disease after intervention].

Objective: To compare the clinical characteristics and long-term prognosis between male and female patients with premature coronary artery disease (PCAD) post coronary intervention, and analyse the risk factors of major adverse cardio-cerebrovascular events (MACCE) and bleeding events. Methods: This was a prospective single-center observational study. From January 2013 to December 2013, 4 744 patients diagnosed as PCAD and treated with percutaneous coronary intervention (PCI) in Fuwai Hospital were enrolled. The general clinical data, laboratory results and interventional treatment data of all patients were collected, and patients were followed up for 2 years after PCI and the incidence of events including MACCE and bleeding was analyzed. The baseline data and clinical events of PCAD patients of different genders were compared. Survival curves were estimated by Kaplan-Meier method. Univariate and multivariate Cox regression were used to analyze whether gender was an influencing factor of different clinical events of PCAD patients within 2 years after PCI, and other relevant influencing factors of MACCE and bleeding events. Results: Among the 4 744 PCAD patients included, there were 3 390 (71.5%) male aged (47.0±5.4) years old and 1 354 (28.5%) female aged (57.0±5.8) years old. Compared with female patients, male patients had higher body mass index, higher proportion of hyperlipidemia, smoking, myocardial infarction, previous PCI, preoperative estimated glomerular filtration rate, ST-segment elevation myocardial infarction, radial artery approach, intravenous ultrasound use and chronic occlusive lesions (all P<0.05). Age, left ventricular ejection fraction, prevalence of hypertension, diabetes mellitus, past stroke history, non-ST-segment elevation acute coronary syndrome (NSTE-ACS) and the use of calcium channel blockers were lower in male patients than in female patients (all P<0.05). The 2-year follow-up results showed that the incidence of BARC type 1 hemorrhage was significantly higher in female patients than in male patients (6.9%(92/1 343) vs. 3.7%(126/3 378), P<0.001); however, the incidence of MACCE, all-cause death, cardiac death, recurrent myocardial infarction, revascularization (target vessel revascularization and target lesion revascularization), stent thrombosis, stroke and BARC type 2-5 hemorrhage were similar between the two groups (all P>0.05). Multivariate Cox regression analysis showed that gender was an independent risk factor for BARC type 1 bleeding in PCAD patients (HR=2.180, 95%CI 1.392-3.416, P<0.001), but it was not an independent risk factor for MACCE and BARC type 2-5 bleeding(all P>0.05). Hyperlipidemia, preoperative SYNTAX score, multivessel lesions and NSTE-ACS were the independent risk factors for MACCE in PCAD patients with PCI (the HRs(95%CI) were 1.289(1.052-1.580), 1.030(1.019-1.042), 1.758(1.365-2.264), 1.264(1.040-1.537), respectively); gender (HR=1.579, 95%CI 1.085-2.297, P=0.017), hyperlipidemia (HR=1.305, 95%CI 1.005-1.695, P=0.046), anticoagulant drugs including low molecular weight heparin (HR=1.321, 95%CI 1.002-1.741, P=0.048) or sulfonate(HR=1.659, 95%CI 1.198-2.298, P=0.002) were the independent risk factors for bleeding events. Conclusions: There are differences in clinical and coronary artery lesion characteristics between different genders in patients with PCAD. The incidence of minor bleeding is significantly higher in female PCAD patients than in male PCAD patients. Hyperlipidemia, preoperative SYNTAX score, multivessel lesions and NSTE-ACS are the independent risk factors for MACCE, and gender, hyperlipidemia, anticoagulant drugs including low molecular weight heparin or sulfonate are the independent risk factors for bleeding events in patients with PCAD.

Effect of estrogen on right ventricular remodeling of monocrotaline-induced pulmonary arterial hypertension in rats and its mechanism.

OBJECTIVE: To elucidate the effect of estrogen on right ventricular remodeling of pulmonary arterial hypertension (PAH) induced by monocrotaline (MCT) in rats and its underlying mechanism. MATERIALS AND METHODS: Male Sprague- Dawley (SD) rats were adaptively fed for one week, and then, randomly divided into control group, MCT group, MCT+17-ß estradiol (50 mg·kg-1·d-1) group, and MCT+17-ß estradiol (100 mg·kg-1·d-1) group, with 8 rats in each group. PAH rat model was constructed by subcutaneous injection of 60 mg·kg-1 MCT for consecutive 4 weeks. The right external jugular vein was intubated to monitor rat RVSP (right ventricular systolic pressure) and mPAP (mean pulmonary arterial pressure). After animal procedures, RV (right ventricle) and LV+S (left ventricle+ventricular septum) of rat were harvested and weighed. Rat tibia was collected and recorded for its length, followed by calculation of RV/(LV+S) and RV/length of the tibia. HE staining and Masson staining were conducted to observe the pathological change and collagen deposition in RV, respectively. RV was prepared for homogenate, followed by detection of total antioxidant capacity (T-AOC) and malondialdehyde (MDA) activities. Expression levels of collagen I, collagen III, NOX4, and NF-κB in RV of PAH rats were detected by qRT-PCR and Western blot. RESULTS: Lower RVSP, mPAP, RV/(LV+S) and RV/length of the tibia were observed in rats of MCT+17-ß estradiol (50 mg·kg-1·d-1) group and MCT+17-ß estradiol (100 mg·kg-1·d-1) group compared with those of MCT group. Injection of 17-ß estradiol (50 mg·kg-1·d-1 and 100 mg·kg-1·d-1) in PAH rats remarkably alleviated pathological changes and collagen deposition in RV. T-AOC activity increased, whereas MDA activity decreased in PAH rats injected with 17-ß estradiol. Expression levels of collagen I, collagen III, NOX4, and NF-κB in RV of PAH rats were all downregulated by 17-ß estradiol treatment. CONCLUSIONS: 17-ß estradiol could remarkably alleviate MCT-induced right ventricular remodeling of PAH rats. It is suggested that 17-ß estradiol exerts its protective role in PAH by inhibiting NOX4-mediated oxidative stress and NF-κB-mediated collagen deposition.

[Impact of short-time anticoagulant therapy after selective percutaneous intervention on prognosis of patients with coronary artery disease].

Objective: To observe the safety and impact of short-term anticoagulant therapy on prognosis after selective percutaneous coronary intervention (PCI) in patients with coronary artery disease. Methods: From January 2013 to December 2013, 9 769 consecutive patients underwent selective PCI in Fuwai Hospital were retrospectively included in this study. Patients were divided into two groups, including non-post-PCI anticoagulant therapy group and low-dose and short-time post-PCI anticoagulant therapy group (enoxaparin 0.4 ml/12 h or fondaparinux 2.5 mg/day by subcutaneous injection for 2-3 days after PCI). All patients were evaluated at 30 days, 180 days and 12 months for major adverse coronary and cerebral events (MACCE) including all-cause death, myocardial infarction, revascularization and stroke as well as in-stent thrombosis and bleeding events. Data from 1 755 pairs of patients were analysis after propensity score matching. The clinical outcomes were compared between groups by using Kaplan-Meier survival analysis before and after propensity score matching. Multivariable Cox analysis was used to define the impact and determinants of post-PCI anticoagulation on clinical outcomes. Results: one thousand seven hundred and fifty-five (18.0%) patients didn't receive post-PCI anticoagulation and 8 014 (82.0%) patients received post-PCI anticoagulation, 5 666 (58.0%) patients received enoxaparin and 2 348 (24.0%) patients received fondaparinux. Patients were younger and incidence of female patients was less, incidence of renal dysfunction and acute coronary syndrome were higher in low-dose and short-time post-PCI anticoagulant therapy group than in non-post-PCI anticoagulation group (all P<0.05). Similarly, patients with post-PCI anticoagulation were associated with more left main coronary artery lesion and branch lesion (P<0.05). Post-PCI anticoagulation patients were associated with less trans-femoral process, more drug-eluting stents implantation and less simple balloon dilatation (all P<0.05). Nine thousand seven hundred and seventeen (99.5%) patients completed 2 years follow up. Post-PCI anticoagulation patients had significantly lower 30-day all-cause death (0.05% (4 cases) vs. 0.46% (8 cases), P<0.001) and stroke (0 vs. 0.11% (2 cases), P=0.003), lower 180-day all-cause death (0.17% (14 cases) vs. 0.57% (10 cases), P=0.002), revascularization (2.07% (166 cases) vs. 3.71% (65 cases), P<0.001) and MACCE (3.49% (280 cases) vs. 5.47% (96 cases), P<0.001), lower 2-year revascularization (7.61% (610 cases) vs. 12.84% (225 cases), P<0.001) and MACCE (10.92 (875 cases) vs. 16.01% (281 cases), P<0.001). Multivariable Cox regression analysis showed that post-PCI anticoagulant therapy was an independent protective factor of 30-day (HR=0.17, 95%CI 0.05-0.62, P=0.007), 180-day all-cause death (HR=0.37, 95%CI 0.16-0.87, P=0.023) and MACCE (HR=0.74, 95%CI 0.58-0.94, P=0.013), 2-year MACCE (HR=0.71, 95%CI 0.62-0.81, P<0.001). After propensity score matching, post-PCI anticoagulation therapy remained as an independent protective factor of 30-day all-cause death (HR=0.11, 95%CI 0.01-0.92, P=0.042) and 2-year MACCE (HR=0.81, 95%CI 0.68-0.96, P=0.015). Conclusions: Low-dose and short-time post-PCI anticoagulant therapy may decrease 30-day all-cause death, 180-day all-cause death and MACCE and 2-year MACCE, and meanwhile this option does not increase bleeding risk in patients underwent selective PCI.

[Pathophysiologic mechanism of CMTM5 low expression in multiple myeloma progression].

Objective: To investigate the mechanism of chemokine-like factor superfamily member (CMTM) 5 on the proliferation of multiple myeloma cells. Methods: RT-qPCR method was used to detect the expression and correlation of CMTM5, caspase3 and caspase9 in U266 after decitabine demethylation treatment; U266 transfected with pcDNA3.1 plasmid overexpressed CMTM5, then cell proliferation activity was detected by CCK-8 assay. Results: Compared with the control group, the low-dose demethylation treatment increased mRNA expression of CMTM5, caspase3, and caspase9 in U266, and showed a time-dependent (P<0.01). The up-trend of CMTM5, caspase3, and caspase9 in the high-demethylation drug treatment group was more significant and also showed time-dependent (P<0.001); There was a significant positive correlation between CMTM5 and caspase3 (r=0.937) and caspase9 (r=0.945) in each group (P<0.001). After transfection of U266 with the pcDNA3.1-CMTM5 plasmid, overexpression of CMTM5 inhibited the cell proliferation activity compared with the control and pcDNA3.1-vector group. Conclusion: Decitabine has a reductive effect on the low level of CMTM5 in U266 cells, and its recovery level is significantly positively correlated with caspase 3 and caspase9. Re-expression of CMTM5 inhibits the proliferative activity of U266.