[摘 要] 目的：探讨m6A调控因子在宫颈癌预后评估、免疫微环境特征及治疗反应中的作用，构建基于m6A调控因子的预后风险模型，并分析其与免疫浸润、药物敏感性及细胞间通信的关系。方法：基于TCGA和GEO数据库中宫颈鳞癌及腺癌（CESC）转录组数据，分析22个m6A调控因子的表达与突变情况；采用LASSO‑Cox回归构建m6A相关预后风险模型，并依据风险评分将患者分为高、低风险组。利用CIBERSORT和ESTIMATE算法评估两组免疫细胞浸润及微环境特征；通过oncoPredict预测抗肿瘤药物敏感性。整合单细胞转录组数据（E-MTAB-12305），采用CellChat分析细胞间通信网络。结果：共鉴定出12个在宫颈癌中差异表达的m6A调控因子。高风险组患者总生存期缩短（P < 0.05），免疫评分降低，且对紫杉醇、氟尿嘧啶、多柔比星等抗肿瘤药物敏感性较差（IC50较高）。单细胞分析显示宫颈癌组织中细胞间通信网络发生改变，其中T细胞与内皮/上皮细胞间的相互作用可能主要由CCL5‑ACKR1和MIF‑（CD74 + CXCR4）配体‑受体介导。在临床样本中，对关键的m6A调控因子进行mRNA和蛋白水平的验证，结果显示部分调控因子在宫颈癌组织中的表达水平高于正常宫颈组织（P < 0.05）。结论：m6A调控因子特征可有效预测宫颈癌患者预后，其风险评分与肿瘤免疫浸润、药物敏感性及细胞通信密切相关，为宫颈癌的预后分层及个体化治疗提供了新的分子依据。

Objective To analyze the effect of immune function on the condition and prognosis of asthma in children with asthma. Methods A total of 148 children with asthma diagnosed in Qinghai women and children's Hospital from January 2018 to January 2021 were included in the analysis, the immune function of the children was determined, and the information of all children was followed up for 6 months after treatment; compared The condition and follow-up prognosis of children with immunocompromised and normal immune function were analyzed and discussed, and the correlation between the expression levels of immunoglobulins (IgG, IgA, IgM) and the condition and short-term recurrence prognosis (6 months) of children was analyzed and discussed, so as to guide Prevention and clinical work. Statistical analysis was done using SPSS19.0. Results The average age of 148 children with recurrent respiratory tract infection in the study was (8.94±3.65) years old, including 70 male children. The condition of the children was evaluated and classified into mild/severe cases: 148 children in this study included mild cases. There were 98 cases and 50 severe cases. There were more males and lower BMI levels in severe children (P<0.05) . The levels of IgG, IgA and IgM in children were all lower in severe children (P<0.05) . The follow-up found that the proportion of relapses in critically ill children was higher (P<0.05). Comparing the levels of IgG, IgA, and IgM in mild and severe children, the average levels of IgG, IgA, and IgM in severe children were lower than those in the mild group, and the difference was statistically significant (P<0.05); recurrence within 6 months of follow-up Prognostic evaluation showed that 19 of the 148 children had relapse, and the levels of IgG, IgA, and IgM in severe relapsed children were significantly lower than those without relapse (P<0.05). Analysis of the relevant factors potentially affecting the prognosis of recurrence showed that gender (female) (OR=1.726) , BMI level (weight loss) (OR=1.613) , IgG expression level factor (low expression) (OR=1.898) , IgA expression Level factor (low expression) (OR=3.509) , IgM expression level factor (low expression) (OR=3.217) and disease factor (severe) (OR=3.619) were potential risk factors, which would increase the risk of poor prognosis. Conclusion The asthma attack in children with immunocompromised immune function is relatively severe, and the short-term recurrence probability is higher, which deserves clinical attention and preventive intervention.

Objective: To explore the hemorrhagic transformation(HT) and its effect on prognosis in patients with acute ischemic stroke(AIS) after mechanical thrombectomy(MT). Methods: A total of 114 patients with AIS received MT were enrolled. The modified Rankin Scale was used to evaluate the clinical outcome at 90 days of onset(0-2 points were good prognosis; 3-6 points were poor prognosis). The patients were divided into HT group(n=25) and non-HT group(n=89) according to their HT conditions. Binomial Logistic regression analysis was performed to determine the vascular risk factors of HT after MT and the effect of HT on prognosis. Results: Among 114 patients, there were 25 cases of HT and 89 cases of non HT. The proportion of patients with diabetes in HT group was significantly higher than that in non-HT group. The NIHSS score of HT group at discharge was significantly higher than that in non-HT group. The proportion with good prognosis at 90 days in HT group was significantly lower than that in non-HT group(allP<0.05). Binomial Logistic regression analysis showed that diabetes, high levels of cholesterol and smoking were the major vascular risk factors for HT after thrombectomy(allP<0.05). HT was an important factor affecting the poor prognosis after arterial thrombectomy(P=0.026). Conclusion Diabetes, high levels of cholesterol and smoking are the main risk factors of HT after MT for AIS. HT is an independent risk factor for poor prognosis after MT.

Objective To analyze the correlation between intestinal flora changes and neonatal necrotizing enterocolitis (NEC)through 16S rRNA metagenomic sequencing and bacterial culture. Methods From September 2018 to March 2019, 10 NEC cases and 6 controls were randomly selected in the neonatal ICU ward of Nanjing maternal and child health care hospital to analyze the 16S rRNA metagenomic diversity of the for intestinal flora. The fecal samples and corresponding environmental samples were corrected from 51 cases of NEC children and their case controls to isolate and culture Clostridium. Results The dispersion of samples within the case group was smaller than that of the control group, and the sample diversity was higher than that of the control group. In the isolation and culture of Clostridium, the overall detection rate of Clostridium in the case group was 43.14% (22/51), and the detection rate of Clostridium butyricum was the highest (19.61%, 10/51). There was a statistical difference between the two groups (χ²=5.85, P=0.015 58). All Clostridium strains did not carry the A, B and E type neurotoxin genes. Conclusion: Increased intestinal flora diversity, intestinal flora abundance and changes in the abundance of Clostridium may be closely related to the intestinal environment of children with NEC; Clostridium, especially Clostridium butyricum, may be related to the occurrence of NEC.

Objective    To compare the efficacy and safety of mitral valvuloplasty via minimally invasive approach with those of mitral valvuloplasty via traditional median sternotomy. Methods    A total of 1 221 patients undergoing mitral valvuloplasty from January 2015 to August 2018 in Guangdong Provincial People&#39;s Hospital were analyzed retrospectively, including 721 males and 500 females, with an average age of 47.2±15.1 years. According to the different surgical methods, they were divided into a study group (n=654), who received mitral valvuloplasty via the totally thoracoscopic approach, and a control group (n=567), who received mitral valvuloplasty via traditional median sternotomy. Clinical data, surgical results, and perioperative outcomes of the two groups were compared. Results    There was no significant difference in preoperative general data between the two groups (P>0.05). Compared with the control group, the study group had longer cardiopulmonary bypass time and aortic cross-clamping time (146.7±42.4 min vs. 122.7±30.6 min, 96.2±32.7 min vs. 78.3±23.8 min, both P=0.000), and shorter total operation time (227.4±55.3 min vs. 238.1±56.4 min, P=0.001). There was no significant difference in the incidence of secondary cross-clamping and mitral valve replacement between the two groups (3.7% vs. 2.6%, P=0.312; 1.7% vs. 1.4%, P=0.690). The blood transfusion rate and the incidence of respiratory tract infection and postoperative poor wound healing were lower (13.0% vs. 24.5%, 2.1%vs. 18.0%, 1.5% vs. 5.3%, all P=0.000) and the postoperative hospital stay was shorter (6.2±4.4 d vs. 11.5±8.8 d, P=0.000) in the study group. There was no significant difference in hospitalization expense between the two groups (95 847.9±31 322.0 yuan vs. 99 673.1±47 930.3 yuan, P=0.149). Within 30 d after surgery, 1 patient died in the study group and 4 patients died in the control group. Before discharge, there were 4 and 5 patients with severe mitral valve regurgitation in the study group and the control group, respectively. Conclusion    Compared with mitral valvuloplasty via traditional median sternotomy, minimally invasive mitral valvuloplasty is superior in shortening operation time and postoperative hospital stay, lowering blood transfusion rate, and reducing postoperative complications, which can achieve better clinical outcomes.

AIM: To discuss the protective effects of betaine-homocysteine methyl transferase(BHMT)on oxidative damaged human lens epithelial cells(HLEC)induced by homocysteine.

METHODS: HLEC were cultured in vitro and then randomly divided into 3 groups. Normal group:normal cultured HLEC; control group: normal cultured HLEC transfected with empty vector; BHMT gene overexpression group(OE): HLEC transfected with BHMT gene overexpression. All groups were cultured in 10% FCS DMEM +5mmol/L Hcy for 16h. After cultured, BHMT mRNA expression was measured by qRT-PCR and Western blot, the cell proliferation was detected by EdU Assay Kit,The level of ROS and GSH of HLEC were measured by Flow Cytometer and Visible Spectrophotometers. The expression level of of protein(GRP78, Nrf2, Caspase-12)was measured by western blotting.

RESULTS: After cultured 16h, cell proliferation ability in OE group was increased by 30.0% compared with NC group(P<0.05).The expression of ROS in normal group(89.2043±0.3511)% was obviously higher than OE group(49.5625±0.4502)%, P<0.05, GSH activity in OE group was obviously higher than control group and normal group,(P<0.05). The expression level of GRP78 in the normal group and the control group was significantly higher than overexpression group. The expression level of Nrf2 in the normal group and the control group was significantly lower than overexpression group. The expression level of Caspase-12 in the overexpression group was significantly lower than that in the control group.

CONCLUSION: BHMT in vitro can prevent the oxidative damage of HLEC by high homocysteine, clear the ROS and decrease the ER stress reaction. Apoptosis of lens epithelial cells was inhibited. BHMT plays an important protective role in oxidative damaged HLEC induced by Hcy.