Necroptotic TNFα-Syndecan 4-TNFα Vicious Cycle as a Therapeutic Target for Preventing Temporomandibular Joint Osteoarthritis

Temporomandibular joint osteoarthritis (TMJOA) is a chronic degenerative disease for which the underlying mechanism still remains unclear. Compared with apoptosis and autophagy, necroptosis causes greater harm to tissue homeostasis by releasing damage-associated molecular patterns (DAMPs). However, the role of necroptosis and downstream key DAMPs in TMJOA is unknown. Here, rodent models of TMJOA were established by the unilateral anterior crossbite (UAC). Transmission electron microscopy (TEM) and immunohistochemistry of receptor interacting protein kinase 3 (RIPK3)/phosphorylation of mixed lineage kinase domain-like protein (pMLKL) were conducted to evaluate the occurrence of necroptosis in vivo. The therapeutic effects of blocking necroptosis were achieved by intra-articularly injecting RIPK3 or MLKL inhibitors and using RIPK3 or MLKL knockout mice. In vitro necroptosis of condylar chondrocyte was induced by combination of tumor necrosis factor alpha (TNFα), second mitochondria-derived activator of caspases (SMAC) mimetics and carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]- fluoromethylketone (z-VAD-fmk). The possible DAMPs released by necroptotic chondrocytes were screened by quantitative proteomics and blocked by specific antibody. Translucent cytosol, swollen organelles, and ruptured cell membranes, features of necroptosis, were frequently manifested in chondrocytes at the early stage of condylar cartilage degeneration in TMJOA, which was accompanied by upregulation of RIPK3/pMLKL. Inhibiting or knocking out RIPK3/MLKL significantly prevented cartilage degeneration. DAMPs released by necroptotic condylar chondrocytes, such as syndecan 4 (SDC4) and heat shock protein 90 (HSP90), were verified. Furthermore, blocking the function of SDC4 significantly attenuated the expression of TNFα in cartilage and synovium, and accordingly increased cartilage thickness and reduced synovial inflammation. Thus, the necroptotic vicious cycle of TNFα-SDC4-TNFα contributes to cartilage degeneration and synovitis, and can serve as a potential therapeutic target for treating TMJOA. © 2022 American Society for Bone and Mineral Research (ASBMR).     

人体测量指标预测肥胖儿童非酒精性脂肪肝的研究

目的 研究人体测量指标对儿童肥胖伴非酒精性脂肪肝的预测作用,探讨不同指标筛查非酒精性脂肪肝的切点值。方法 选取自2018年6月—2019年12月在西安交通大学第二附属医院小儿内分泌门诊就诊的94例肥胖儿童为研究对象,进一步分为肥胖伴非酒精性脂肪肝组与肥胖不伴非酒精性脂肪肝组,52例正常儿童为对照组。测量身高(H)、体重(W)、腰围(WC)、臀围(HC)和血甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C);行肝脏B超的检查。计算体质指数(BMI)、腰臀比(WHR)、腰高比(WHtR)、腹部体积指数(AVI)、脂质蓄积指数(LAP)和内脏脂肪指数(VAI)等指标。通过绘制人体测量指标的受试者工作特征曲线(ROC曲线)评估人体测量指标与肥胖儿童非酒精性脂肪肝的相关性,并比较各项人体测量指标的曲线下面积(AUC)确定切点值。结果 肥胖组BMI、WC、WHR、WHtR、AVI、LAP、VAI及TG均高于对照组(t=23.090、21.068、12.547、22.855、17.578、8.159、5.394、6.183,P＜0.001)。肥胖伴非酒精性脂肪肝组BMI、WC、AVI、LAP、VAI均高于肥胖不伴非酒精性脂肪肝组(t=2.180、2.389、2.362、3.643、2.839,P＜0.05)。人体测量指标的ROC曲线下面积按从大到小的顺序依次为 LAP、VAI、 WC、 AVI、 BMI。对肥胖伴非酒精性脂肪肝的联合诊断指标进行筛查效能分析结果显示,LAP+AVI 曲线下面积为0.706(95%CI:0.595~0.817,P＜0.001);AVI+VAI 曲线下面积为0.685(95%CI:0.570~0.800,P＜0.01);BMI+WC 曲线下面积为0.652(95%CI:0.537~0.768,P＜0.05)。联合指标的ROC曲线下面积从大到小为LAP+AVI、AVI+VAI、BMI+WC。结论 LAP联合AVI对儿童肥胖伴非酒精性脂肪肝具有较好的筛查作用。     

重点部门常见典型操作时的高频接触表面

目的 明确不同部门的高频接触表面,为清洁消毒工作提供依据与指导。方法 采用直接观察法对西安市某三级甲等综合医院的消化内镜室、口腔科、血透室、手术室、检验科5个医院感染管理重点部门环境表面的接触频次进行调查,计算累积接触频次与频率。结果 5个重点部门高频接触表面均为5~9个。消化胃镜室高频接触表面频次（3.57~13.29次/操作）明显多于其他部门;血透室高频接触频次范围为0.56~3.44次/操作,为5个部门中最低者。接触频次最高的表面：消化内镜室为诊疗床（13.29次/操作）,口腔科为综合治疗台操作面板（3.64次/操作）,血透室为透析机触屏（3.44次/操作）,手术室为麻醉用电脑键盘及鼠标（8.25次/操作）,检验科为电脑键盘及鼠标（4.80次/操作）。治疗车为消化内镜室、手术室及血透室的高频接触表面。结论 因医疗诊疗操作的不同,各部门高频接触表面也不尽相同。电脑鼠标和键盘、治疗车、诊疗床、诊疗专用仪器以及操作台面在调查的各重点部门均为高频接触表面。     

西安市134例百日咳疑似病例不同标本百日咳鲍特菌检测阳性率比较

目的比较百日咳疑似病例鼻、咽和鼻咽拭子标本百日咳鲍特菌(Bordetella pertussis,Bp)检测阳性率。方法选择西安市两家医院2018年1-7月134例百日咳疑似病例,同时采集鼻、咽和鼻咽拭子标本,进行Bp分离培养和实时荧光定量聚合酶链反应核酸检测。结果百日咳疑似病例的Bp检测总阳性率为44.78%(60/134),其中鼻、咽、鼻咽拭子阳性率分别为36.57%、32.09%、39.55%(χ²=1.64,P=0.440);发病后0-7d、8-14d、15-21d、22-28d、29d采样的阳性率分别为23.33%、64.52%、63.89%、42.86%、23.33%(χ²=22.13,P=0.000)。鼻、咽、鼻咽拭子Bp核酸检测阳性率分别为36.57%、32.09%、39.55%(χ²=1.64,P=0.440);分离培养阳性率分别为0.00%、0.00%、4.59%(Fisher精确概率法,P=0.036)。在Bp核酸阳性病例中鼻和/或咽、鼻和/或鼻咽、咽和/或鼻咽拭子阳性分别占93.33%、91.67%、100%(Fisher精确概率法,P=0.080)。结论百日咳疑似病例鼻、咽和鼻咽拭子Bp检测总阳性率相当;核酸检测阳性率以鼻咽拭子最高,组合采样可提高Bp核酸检测阳性率;分离培养仍建议选择鼻咽拭子。     

重症先天性心脏病手术期间氧供量和氧耗量的变化

目的了解重症先天性心脏病手术期间的氧合状态。方法测定了32例心脏复跳后连续应用多巴酚丁胺或米力农情况下体外循环中、停体外循环后30min、手术结束、术后2h、术后16h等时点的心脏指数(CI)、氧供量(DO2)、氧耗量(VO2)和氧摄取率(ERO2)。结果①体外循环中和体外循环结束DO2和VO2有高度正相关性(P<0.01),相关系数分别为0.861,0.811;②体外循环中与体外循环后30min各数据比较CI、DO2、VO2前者明显低于后者(P<0.05或0.01),ERO2无显著性差异;③体外循环结束后各点数据比较,CI能维持在3L/(min·m2)以上,DO2能维持在550ml/(min·m2)以上。VO2能维持在120ml/(min·m2)以上。结论重症先天性心脏病手术期间存在病理性氧供依赖。心脏复跳后应用多巴酚丁胺能改善组织对氧的摄取和利用。体外循环后连续应用多巴酚丁胺或米力农很难进行DO2、VO2、CI的超正常值维护,但CI不小于3L/(min·m2)。手术后16h循环功能尚未完全恢复,仍需继续加强正性肌力药的治疗。     

乙型肝炎病毒表面抗原(HBsAg)基因重组杆状病毒的构建及鉴定

目的 利用Bac-to-Bac杆状病毒表达系统，构建乙型肝炎病毒(HBV)表面抗原基因的重组杆状病毒。方法 构建含有HBV S基因的表达截体pFBDHTa—S，转化DH10Bac感受态菌。利用其含有的细菌Tn7转座系统，将该基因重组至杆状病毒穿梭质粒bacmicl中，转染昆虫细胞Sf9，获得含有HBV S基因的重组杆状病毒。结果 构建了的含HBV S基因的重组杆状病毒，感染昆虫细胞后，PCR检测可以扩增出相应大小的片段。结论 利用杆状病毒表达系统，成功地构建了含HBV S基因的重组杆状病毒，为进一步的研究奠定了基础。     

视觉电生理和视野检查对原发性开角型青光眼早期诊断的敏感性

目的:探讨多种视觉电生理联合全自动视野计检查对原发性开角型青光眼早期诊断中的敏感性,为临床早期诊断提供客观敏感的指标。方法:选择10例(20眼)原发性开角型青光眼患者和正常对照者10例(20眼)分别进行闪光视网膜电图(FERG)、图形视网膜电图(PERG)、图形视觉诱发电位(PVEP)检查;还对正常对照组、青光眼组中各5例(10眼)进行多焦视网膜电图(mERG)检查;青光眼组10例(20眼)和正常对照组8例(16眼)行蓝黄视野(B/Y)、标准视野(W/W)检查。结果:PERG的异常率较高,PERG的潜伏期延长较敏感,mERG二阶反应黄斑区振幅下降,随离心度的增大,振幅逐渐变小,潜伏期延长,PVEP的P100波振幅降低,潜伏期延长。B/Y视野异常与W/W视野异常比较有显著差异(P <0.01)。结论:B/Y视野检查联合多种视功能检查对原发性开角型青光眼早期诊断较敏感。早期诊断中的B/Y视野敏感性高;若眼压差大,眼底C/D未出现改变,则PERG可作为较敏感观测指标。     

Optimization of voriconazole dosage regimen to improve the efficacy in patients with invasive fungal disease by pharmacokinetic/pharmacodynamic analysis

Invasive fungal disease (IFD) is a significant cause of morbidity and mortality in hospitalized patients. To maximize the efficacy of voriconazole treatment, the study established the relationship between voriconazole pharmacokinetic/pharmacodynamic (PK/PD) and probability of response and optimized voriconazole dosage regimen in patients with IFD based on Monte Carlo simulation. Forty‐four patients proven with IFD were involved in this study. Among them, the overall cure rate was 75% (33/44) and there was a significant difference between C_min/MIC values in patients with lack of response (n = 11) and those with successful response (n = 33) (mean value: 1.91 vs. 11.33; P < 0.05). Logistic regression model showed a high correlation between voriconazole C_min/MIC ratio and clinical response (P = 0.044, OR = 1.349). According to Monte Carlo simulation results under different voriconazole dosing regimens, we could draw a conclusion that 200 mg voriconazole administered intravenously or orally twice daily for Candida infections and 300 mg administered orally or with 200 mg administered intravenously twice daily for Aspergillus infections were rational, which could achieve a value of the cumulative fraction of response >90%. This study built the relationship between voriconazole PK/PD and clinical response and obtained the reasonable empirical dosage regimen, which can be used to customize individual dosage regimen and improve the efficacy of voriconazole treatment.     

Activation of G protein-coupled receptor 30 protects neurons by regulating autophagy in astrocytes

Ischemic stroke leads to neuronal damage induced by excitotoxicity, inflammation, and oxidative stress. Astrocytes play diverse roles in stroke and ischemia-induced inflammation, and autophagy is critical for maintaining astrocytic functions. Our previous studies showed that the activation of G protein-coupled receptor 30 (GPR30), an estrogen membrane receptor, protected neurons from excitotoxicity. However, the role of astrocytic GPR30 in maintaining autophagy and neuroprotection remained unclear. In this study, we found that the neuroprotection induced by G1 (GPR30 agonist) in wild-type mice after a middle cerebral artery occlusion was completely blocked in GPR30 conventional knockout (KO) mice but partially attenuated in astrocytic or neuronal GPR30 KO mice. In cultured primary astrocytes, glutamate exposure induced astrocyte proliferation and decreased astrocyte autophagy by activating mammalian target of rapamycin (mTOR) and c-Jun N-terminal kinase (JNK) and inhibiting p38 mitogen-activated protein kinase (MAPK) pathway. G1 treatment restored autophagy to its basal level by regulating the p38 pathway but not the mTOR and JNK signaling pathways. Our findings revealed a key role of GPR30 in neuroprotection via the regulation of astrocyte autophagy and support astrocytic GPR30 as a potential drug target against ischemic brain damage.     

索拉菲尼治疗进展期原发性肝癌患者有效性及安全性研究*

目的 探讨应用索拉菲尼治疗进展期原发性肝癌(aPLC)患者的有效性和安全性。方法 2013年4月~2016年12月我科诊治的aPLC患者82例,采用随机数字表法分为两组,每组41例。两组均给予射频消融术（RFA）治疗,观察组患者在RFA前后接受索拉菲尼治疗,观察12 w。采用ELISA法检测血清碱性成纤维细胞生长因子（bFGF）和血管内皮生长因子（VEGF）。结果 2例观察组患者被剔除,3例对照组患者失访;在治疗12 w末,观察组疾病控制率为61.5%,与对照组的52.6%比,差异无统计学意义（P>0.05）,但观察组肿瘤客观有效率为48.7%,显著高于对照组的26.3%,差异有统计学意义（P<0.05）;观察组血清AFP、bFGF和VEGF水平分别为（184.7±10.5）μg/L、（3.8±1.3） pg/mL和（172.3±25.4） pg/mL,均显著低于对照组的（213.6±11.6） μg/L、（6.4±2.0） pg/mL和（210.5±28.3） pg/mL,差异有统计学意义（P<0.05）;观察组中位无进展生存期（PFS）为10.2个月（95%CI为7.4~11.5）,对照组为7.9个月（95%CI为 6.0~10.1）,经Log-rank检验显示两组差异有统计学意义（P<0.05）;观察组在服药过程中手足综合征、皮疹、白细胞减少、口腔黏膜炎、脱发和肝功能异常发生率分别为43.6%、25.6%、17.9%、20.5%、25.6%和23.1%,显著高于对照组的0.0%、2.6%、0.0%、0.0%、0.0%和5.3%,差异有统计学意义（P<0.05）。结论 索拉菲尼可控制aPLC患者实体瘤扩散,延长无进展生存期,但索拉菲尼可引起多种不良反应,在用药过程中应注意观察并及时采取相应处理措施,以防止发生严重不良反应。