目的 比较规律口服中药、未规律口服中药、规律口服中药+中医非药物治疗3种方案对新型冠状病毒奥密克戎变异株感染者的疗效.方法 回顾性分析重庆援吉医疗队2022年4月6日至2022年4月18日在长春奢岭方舱医院收治的766例新型冠状病毒奥密克戎感染患者,按其实际接受的治疗方法分为规律口服中药、未规律口服中药、规律口服中药+中医非药物治疗组.规律口服中药组(n=337)每日3次口服中药治疗,未规律口服中药组(n=152)间断口服中药治疗,规律口服中药+中医非药物治疗组(n=277)在每日3次规律口服中药的基础上配以耳穴、安眠包和穴位敷贴.比较各组患者出院达标时间.结果 规律口服中药组、规律口服中药+中医非药物治疗组与未规律口服中药组比较,出院达标时间差异具有统计学意义(P＜0.05);规律口服中药+中医非药物治疗组与规律口服中药组比较,出院达标时间差异无统计学意义(P＞0.05).结论 规律口服中药可促进患者核酸转阴,缩短住院治疗时间.

Introduction: Primary brainstem hemorrhage (PBSH) is the most fatal subtype of intracerebral hemorrhage and associated with poor prognosis. We aimed to develop a prediction model for predicting 30-day mortality and functional outcome in patients with PBSH. Methods: We reviewed records of 642 consecutive patients with first-time PBSH from three hospitals between 2016 and 2021. Multivariate logistic regression was used to establish a nomogram in a training cohort. Cutoff points of the variables were determined by receiver operating characteristic curve analysis, and certain points were assigned to these predictors to produce the PBSH score. The nomogram and PBSH score were compared with other scoring systems of PBSH. Results: Five independent predictors, comprised of temperature, pupillary light reflex, platelet-to-lymphocyte ratio (PLR), Glasgow Coma Scale (GCS) score on admission, and hematoma volume, were incorporated to construct the nomogram. The PBSH score consisted of 4 independent factors with individual points assigned as follows: temperature, ≥38℃(=1 point), <38℃(=0 points); pupillary light reflex, absence (=1 point), presence (=0 points); GCS score 3 to 4 (=2 points), 5 to 11 (=1 point), and 12 to 15 (=0 points); PBSH volume >10 mL (=2 points), 5 to 10 mL (=1 point), and <5 mL (=0 points). Results showed that the nomogram was discriminative in predicting both 30-day mortality [AUC of 0.924 in the training cohort, and 0.931 in the validation cohort] and 30-day functional outcome [AUC of 0.887]. The PBSH score was discriminative in predicting both 30-day mortality [AUC of 0.923 in the training cohort, and 0.923 in the validation cohort] and 30-day functional outcome [AUC of 0.887]. The prediction performances of the nomogram and the PBSH score were superior to the intracranial hemorrhage (ICH) score, primary pontine hemorrhage (PPH) score, and new PPH score. Conclusions: We developed and validated two prediction models for 30-day mortality and functional outcome in patients with PBSH. The nomogram and PBSH score could predict 30-day mortality and functional outcome in PBSH patients.

患者 ,男 ,51 岁 ,因"双下肢结节 、脓肿 1 个月 ,破溃 2 周"入院 . 入院后患者血压下降 ,体温测不出 ,呼吸急促 、意识改变 、面色苍白 、大汗淋漓 ,考虑脓毒症休克转入 ICU . 查体 :体温 38 .0 ℃ ,脉搏 89 次/min ,呼吸 25 次/min ,血压 94/60 mmHg(1 mmHg = 0 .133 kPa)(去甲肾上腺素 0 .3 μg · kg- 1 · min- 1 静脉泵入). 心肺查体阴性 ,左上腹稍压痛 ,腹肌稍紧张 ,肝脾叩大 ,无明显叩击痛 ,左肾区轻叩痛 ,右肾区叩痛阴性.

Objective To investigate the effect of interleukin 6 (IL-6) on autophagy and apoptosis of human umbilical vein endothelial cells (HUVECs) and the interaction between autophagy and apoptosis. Methods HUVECs were cultured in vitro and stimulated with 5, 10, 20, 50 ng/L IL-6 for 24 h. The cell proliferation was detected by cell counting Kit-8 (CCK-8) assay, the apoptosis was detected by flow cytometry, the acidic vesicle organelles were observed by monodansylcadaverine staining, the ultra-microstructural cellular structure was observed by transmission electron microscope, and the expressions of apoptosis-related protein cleaved caspase-3 and autophagy-related proteins, including microtubule-associated protein 1 light chain 3 (LC3) and p62, were detected by Western blotting. HUVECs were stimulated with IL-6 combined with autophagy inhibitor 3-methyladenine (3-MA) or autophagy inducer sirolimus (SRL), and then the changes of cell proliferation, cell apoptosis and expressions of the above proteins were analyzed. Results Compared with the control group, the cell proliferation and the ratio of LC3Ⅱ/LC3Ⅰin HUVECs in the IL-6 groups were significantly decreased (P<0.05), the apoptosis and the expressions of cleaved caspase-3 and p62 were significantly increased (P<0.05), the apoptotic vacuole number was significantly decreased (P<0.05), and the autophagy was reduced. The above effects were concentration-dependent (P<0.05). Compared with the IL-6 group, the autophagy of HUVECs in the IL-6 combined with 3-MA group was inhibited, the numbers of autophagosome and apoptotic vacuole were significantly decreased (P<0.05), LC3Ⅱ/LC3Ⅰratio was significantly decreased (P<0.05), and the cell apoptosis and the expression of cleaved caspase-3 and p62 were significantly increased (P<0.05). Compared with the IL-6 group, the autophagy of HUVECs in the IL-6 combined with SRL group was promoted, the numbers of autophagosome and apoptotic vacuole were significantly increased (P<0.05), LC3Ⅱ/LC3Ⅰratio was significantly increased (P<0.05), and the cell apoptosis and the expression of cleaved caspase-3 and p62 were significantly decreased (P<0.05). Conclusion IL-6 can induce the apoptosis of HUVECs by inhibiting autophagy.

Objective To explore the effect of ambroxol hydrochloride combined with budesonide on the airway inflammation and lung oxidative stress in allergized rats.Methods Forty cases of clean health level SD male rats were randomly divided into blank control group,model group,positive control group and test group,10 rats in each group.Except blank control group,the rats of other groups were treated with ovalbumin to build the asthma model.Model group was intraperitoneally injected 10％ ovalbumin 1 mL on 1,8 d,and 1％ ovalbumin was inhaled on 15 d,once a day for 0.5 h,till the rats appear abdominal muscle contraction,nodding and so on.Positive control group was inhaled 0.02％ budesonide 30 min before nebulization for 1 h before nebulization.The test group was intraperitoneal injected ambroxol hydrochloride 60 mg · kg-1 at 25 min before inhalation of budesonide on the basis of positive control group.Each group were intervention for 10 weeks.The numbers of lymphocytes and eosinophils in bronchoalveolar lavage fluid of rats in each group were measured by cell counting method.The airway histopathological and morphological changes were observed by hematoxylin-eosin (HE) staining.Colorimetric method was used to detect the levels of oxidative stress in lungs of each group.Results The percentages of eosinophils in blank control group,model group,positive control group and test group were (1.29 ±0.41)％,(81.44 ±5.15)％,(37.51 ±5.38)％,(20.14±4.65)％,the percentages of lymphocytes were (4.01 ± 0.87) ％,(17.38-± 2.99) ％,(11.59 ± 2.01) ％,(7.96 ± 1.43) ％.The difference betweent test group and model group and positive control group was statistically significant (P ＜0.01).HE staining showed that the most obvious pathological changes in model group,followed by positive control group.The levels of glutathione (GSH) in lung tissue of blank control group,model group,positive control group and test group were (1.54±0.15),(0.53 ±0.18),(0.79 ±0.09),(1.15 ±0.11) μmol · L-1,and reactive oxygen species (ROS) were (1643 ±163),(4986 ±195),(3619 ± 184) and (2348 ± 171) U mg-1 pro.The GSH,ROS in test group and positive control group had significant difference (P ＜ 0.01).Conclusion Ambroxol hydrochloride combined with budesonide can relieve the airway inflammation and lung oxidative stress in rats with asthma,which can be used for the treatment of acute asthma attacks.

Objective To explore the mechanism by which peroxisome proliferators-activated receptor-γ (PPAR γ) upregulates miR-16 and inhibits inflammatory response in sepsis.Methods Real-time PCR was used to investigate the expression of PPAR γand miR-16 in peripheral blood monocytes of patients with sepsis and healthy subjects,and the correlation between PPAR γand miR-16 was analyzed.THP-1 and RAW246.7 cells were treated with the PPAR γagonist RGZ,PPAR γsiRNA or a mir-16 inhibitor (antagomir16),and the changes in the expressions of miR-16 and its target gene IKKα were detected using real-time PCR and Western blotting.The cells were transfected with luciferase reporter gene plasmid containing the miR-16 promoter region followed by treatment with RGZ or the PPAR γ antagonist GW9662,and luciferase reporter assay was performed to detect the changes in the reporter gene activity.In cells treated with RGZ followed then by LPS,the expression levels of the inflammatory factors tumor necrosis factor α (TNFα) and interleukin-6 (IL-6) were detected using ELISA.In a mouse model of LPS-induced sepsis,following treatment with RGZ with or without antagomir-16 pretreatment,the expression of miR-16 in peripheral blood monocytes was detected with real-time PCR,and the expression of TNFα and IL-6 were determined using ELISA.Results The expressions of PPAR γ and miR-16 in peripheral blood monocytes were significantly reduced in patients with sepsis (P ＜ 0.05).In THP-1 and RAW246.7 cells,the activation of PPAR γ obviously increased the expression of miR-16 by enhancing the transcriptional activity of miR16 promoter and consequently inhibited the expression of IKKα,the target gene of miR-16 (P ＜ 0.05).PPAR γ inhibited production of TNFα and IL-6 in the inflammatory cells by increasing the expression of miR-16 (P ＜ 0.05).Treatment with PPAR γsignificantly decreased the serum levels of TNFα and IL-6 in mouse models of LPS-induced sepsis (P ＜ 0.05).Conclusion Activation of PPAR γ inhibits the expression of inflammatory cytokines in inflammatory cells and suppresses inflammatory response in septic mice by upregulating miR-16.

// Yue Wu 1,* , Jiangmin Wan 1,2,* , Xuemei Chen 3,* , Wei Hua 4 , Haibo Gui 3 , Xushun Jiang 1 , Huizhe Huang 5 , Xiongzhong Ruan 6,7 and Xiaogang Du 1,8 1 Department of Nephrology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China 2 Department of Nephrology, People’s Hospital of Qijiang Distract, Chongqing, China 3 Emergency Department, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China 4 Department of Nephrology, The Sixth People’s Hospital of Chongqing, Chongqing, China 5 The second Affiliated Hospital of Chongqing Medical University, Chongqing, China 6 Centre for Nephrology, Royal Free and University College Medical School, University College London, Royal Free Campus, London, United Kingdom 7 Centre for Lipid Research, Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing, China 8 The Chongqing Key Laboratory of Translational Medicine in Major Metabolic Disease, Chongqing, China * These authors contributed equally to this work Correspondence: Xiaogang Du, email: dxgcxm@163.com Keywords : parkin; mitophagy; palmitic acid; tubular epithelial cells; apoptosis Received: October 07, 2016 Accepted: December 18, 2017 Epub: December 26, 2017 Abstract Chronic kidney disease (CKD) is often accompanied by dyslipidemia. Excessive lipid accumulation in renal tubular epithelial cells (HK-2 cells) triggers mitochondrial dysfunction and overproduction of reactive oxygen species (ROS). Mitophagy reportedly plays a crucial protective role by selectively degrading dysfunctional or damaged mitochondria. This is largely regulated by Parkin, an E3 ubiquitin ligase. In this study, we found increased Parkin, Beclin1 and LC3 expression in the renal tissues of CKD patients and rats fed a high-fat diet (HFD), and in HK-2 cells stimulated by palmitic acid (PA). We also found that PA induced upregulation and translocation of Parkin to mitochondria and increased the colocalization of Parkin and GFP-LC3 puncta, Parkin siRNA reversed this increase, indicating that Parkin participated in PA-induced mitophagy in HK-2 cells. Furthermore, our study demonstrated that PA stimulation decreased MitoTracker Red fluorescence and mitochondrial membrane potential and increased apoptosis and mitochondrial ROS generation in HK-2 cells. However, Parkin deficiency further reduced mitochondrial membrane potential, enhanced apoptosis and production of mitochondrial ROS. These data provide evidence that Parkin-mediated mitophagy plays a protective role against fatty acid-induced renal tubular epithelial cells apoptosis by reducing oxidative stress.

Autophagy is a highly conserved degradation process that is involved in the clearance of proteins and damaged organelles to maintain intracellular homeostasis and cell integrity. Type 2 diabetes is often accompanied by dyslipidemia with elevated levels of free fatty acids (FFAs). Podocytes, as an important component of the filtration barrier, are susceptible to lipid disorders. The loss of podocytes causes proteinuria, which is involved in the pathogenesis of diabetic nephropathy. In the present study, we demonstrated that palmitic acid (PA) promoted autophagy in podocytes. We further found that PA increased the production of reactive oxygen species (ROS) in podocytes and that NAC (N-acetyl-cysteine), a potent antioxidant, significantly eliminated the excessive ROS and suppressed autophagy, indicating that the increased generation of ROS was associated with the palmitic acid-induced autophagy in podocytes. Moreover, we also found that PA stimulation decreased the mitochondrial membrane potential in podocytes and induced podocyte apoptosis, while the inhibition of autophagy by chloroquine (CQ) enhanced palmitic acid-induced apoptosis accompanied by increased ROS generation, and the stimulation of autophagy by rapamycin (Rap) remarkably suppressed palmitic acid-induced ROS generation and apoptosis. Taken together, these in vitro findings suggest that PA-induced autophagy in podocytes is mediated by ROS production and that autophagy plays a protective role against PA-induced podocyte apoptosis.

目的:研究脓毒症患者外周血T淋巴细胞亚群和T淋巴细胞凋亡的变化,探讨其在免疫抑制和预后中的作用.方法:收集2014年10月至2015年12月重庆医科大学附属第一医院ICU收治的脓毒症患者55例,随访患者28 d生存率,根据预后不同,分为脓毒症患者存活组和脓毒症患者死亡组;同时收集健康志愿者30例为对照组.应用流式细胞仪检测患者外周血T细胞亚群计数,分析其与APACHEⅡ评分及预测死亡危险率(R)的相关性;用免疫磁珠分选外周血CD3+T淋巴细胞,通过流式细胞技术和Tunel法检测细胞凋亡.用脓毒症患者和健康志愿者血清体外刺激Jurkat细胞24 h,采用流式细胞技术和Tunel染色法检测细胞凋亡,Western blot法检测激活型天冬氨酸特异性半胱氨酸蛋白酶(cleaved-easpase 3),B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)和髓样细胞白血病-1(myeloid cell leukemia l,Mcl-1)蛋白表达水平.结果:脓毒症患者存活组、死亡组T淋巴细胞各亚群绝对值计数均明显低于对照组,分别为:CD3+ 374.0(274.5～514.0)、154.0(87.5～256.2)、1534.5(1255.0～1756.7),CD3 +CD4+ 198.0(148.5 ～299.0)、68.0(37.7～108.5)、766.5 (629.3～923.5),CD3+CD8+ 142.0 (89.0～184.0)、76.0(48.7～134.5)、600.5(400.2～717.0)(全部P＜0.05);脓毒症患者死亡组CD3+、CD3+CD4+细胞计数减少均较脓毒症患者存活组更显著,差异具有统计学意义(P=0.011,P=0.005).脓毒症患者死亡组外周血T淋巴细胞中CD3+CD4+细胞占(42.40±5.71)％,较对照组(53.46±9.28)％明显减少(P=0.003);CD3+CD8+细胞占(54.40±4.76)％,较对照组(39.33±7.26)％增加(P=-0.003);CD4+/CD8+比值为(0.79±0.17),较对照组(1.44±0.51)明显下降(P=0.023).CD3+、CD3+CD4+、CD3+CD8+、CD4+/CD8+比值与患者APACHEⅡ评分和R均呈负相关关系(全部P＜0.05).脓毒症患者存活组、死亡组CD3+T淋巴细胞凋亡率分别为(15.59±0.54)％、(39.57±4.49)％,较对照组(9.53±1.91)％明显增加(P=0.040,P=0.000),脓毒症患者死亡组较脓毒症患者存活组增加更明显(P=0.000).脓毒症患者存活组、死亡组血清诱导Jurkat细胞凋亡率分别为(16.60±3.55)％、(32.93±5.89)％,较对照组(8.56±0.89)％明显增加(P=-0.049、P=-0.000);Cleaved-caspase3蛋白水平(2.01±0.21,2.68±0.28)较对照组(1.00±0.00)上调(P=-0.001、P=0.000);抗凋亡蛋白Mcl-1(0.77±0.03,0.61±0.01)、Bcl-2 (0.68±0.07,0.48±0.03)较对照组(1.00±0.00)明显下调(P=0.000、P=0.000) (P=0.000、P=0.000);与脓毒症患者存活组比较,脓毒症患者死亡组中Jurkat细胞凋亡、Cleaved-caspase3、Mcl-1、Bel-2变化更加为明显(P=0.002,P=0.007、P=0.000、P=0.001).结论:脓毒症患者免疫失衡,T淋巴细胞凋亡增加,T淋巴细胞数明显减少,发生免疫抑制,且与病情严重程度和预后密切相关.

目的 测定T、B淋巴细胞亚群在急性胰腺炎(acute pancreatitis,AP)患者中的水平,探讨其与AP病情的相关性.方法 收集2013-04～ 2015-04重庆医科大学附属第一医院收治的AP患者200例作为病例组,并选择同期年龄、性别相近的健康体检者40例作为对照组.将AP患者分为轻症急性胰腺炎(mild acute pancreatitis,MAP)组(78例)、中重症急性胰腺炎(moderately severe acute pancreatitis,MSAP)组(73例)及重症急性胰腺炎(severe acute pancreatitis,SAP)组(49例).应用流式细胞学技术分析T淋巴细胞亚群CD3+、CD4+、CD8+,CD4+ CD25+调节性T细胞(regulatory T cell,Treg),以及调节性B淋巴细胞(regulatory B cell,Breg)的表达;应用酶联免疫吸附法(ELISA)测定IL-10和IL-4水平.结果 MAP组、MSAP组及SAP组T淋巴细胞亚群CD3+、CD4+表达率及Breg表达率均低于健康对照组,且以SAP组及MSAP组下降更为明显,差异有统计学意义(P＜0.05);MAP组、MSAP组及SAP组CD4+/CD8+比值＜1.5(分别为41.0％、41.0％、63.3％)和比值＜1.0(分别为20.5％、31.5％、38.8％)的发生率均高于对照组(分别为22.5％和10.0％),差异有统计学意义(P＜0.05).CD4+ CD25+ Treg表达率均明显升高,以SAP组升高最为明显(P＜0.05).AP患者外周血CD3+、CD4+表达率及Breg与Ranson评分、APACHEⅡ评分及SOFA评分呈负相关性(P＜ 0.05);AP患者CD4+ CD25+ Treg表达率与Ranson评分、APACHEⅡ评分及SOFA评分呈正相关性(P＜ 0.05).结论 T、B淋巴细胞亚群改变与AP疾病病程和分级相关.

Background Hyperlipidemia-induced apoptosis mediated by fatty acid translocase CD36 is associated with increased uptake of ox-LDL or fatty acid in macrophages, hepatocytes and proximal tubular epithelial cells, leading to atherosclerosis, liver damage and fibrosis in obese patients, and diabetic nephropathy (DN), respectively. However, the specific role of CD36 in podocyte apoptosis in DN with hyperlipidemia remains poorly investigated. Methods The expression of CD36 was measured in paraffin-embedded kidney tissue samples (Ctr = 18, DN = 20) by immunohistochemistry and immunofluorescence staining. We cultured conditionally immortalized mouse podocytes (MPC5) and treated cells with palmitic acid, and measured CD36 expression by real-time PCR, Western blot analysis and immunofluorescence; lipid uptake by Oil red O staining and BODIPY staining; apoptosis by flow cytometry assay, TUNEL assay and Western blot analysis; and ROS production by DCFH-DA fluorescence staining. All statistical analyses were performed using SPSS 21.0 statistical software. Results CD36 expression was increased in kidney tissue from DN patients with hyperlipidemia. Palmitic acid upregulated CD36 expression and promoted its translocation from cytoplasm to plasma membrane in podocytes. Furthermore, palmitic acid increased lipid uptake, ROS production and apoptosis in podocytes, Sulfo-N-succinimidyloleate (SSO), the specific inhibitor of the fatty acid binding site on CD36, decreased palmitic acid-induced fatty acid accumulation, ROS production, and apoptosis in podocytes. Antioxidant 4-hydroxy-2,2,6,6- tetramethylpiperidine -1-oxyl (tempol) inhibited the overproduction of ROS and apoptosis in podocytes induced by palmitic acid. Conclusions CD36 mediated fatty acid-induced podocyte apoptosis via oxidative stress might participate in the process of DN.

Objective The aim of this study was to investigate the significance of myocardial injury markers combined with APACHEⅡscore in predicting the prognosis of patients with septic shock. Methods Clinical data of 82 patients with septic shock were analyzed retrospectively.The serum concentration of MYO, CK-MB, NT-proBNP, cTnT in patients with septic shock were measured on the day of definite diagnosis.The APACHEⅡscore was calculated and the risk of death was predicted according to APACHEⅡ score, and the survival rate of 28 days were followed up.The relationship between MYO, CK-MB, NT-proBNP, cTnT level and prognosis was analyzed.Results In patients with septic shock, 45.1% of patients had elevated serum CK-MB, 63.4% of patients had elevated&nbsp;serum cTnT, 78.0%of patients had elevated serum MYO, 93.9%of patients had elevated serum NT-proBNP.Compared with survival group, serum levels of MYO, CK-MB and cTnT were higher in death group (P<0.05).In patients with MODS, the serum levels of MYO, CK-MB, NT-proBNP and mortality were higher than those in patients without MODS (P<0.05).MYO, CK-MB, cTnT and APACHEⅡscore had statistical significance in predicting death ( area under ROC curve was 0.673, 0.703, 0.719 and 0.851 respectively, P<0.05).The area under ROC curve of MYO, CK-MB, cTnT and NT-proBNP to predict death together was 0.749 (P<0.05).The area under ROC curve was 0.874 (P<0.05) when MYO, CK-MB, cTnT and NT-proBNP were combined with APACHEⅡscore, which was higher than APACHEⅡscore or myocardial injury markers alone in predicting death. Conclusion Serum levels of myocardial injury markers were higher in death group and group with MODS.MYO, CK-MB and cTnT have predictive effect for the prognosis of patients with septic shock. High serum MYO, CK-MB, cTnT predicts higher mortality in patients with septic shock.Myocardial injury markers combined with APACHEⅡ score have important predictive value for the prognosis of patients with septic shock.

Both Pax1 and Pax9 belong to the important paired box gene family (PAX), which mainly participates in animal development and sclerotome differentiation. To date, the precise molecular mechanism and related signaling pathway of Pax1 remain unclear. In our study, microinjection of morpholino- (MO-) modified antisense oligonucleotides against pax1b induced pectoral fin bud defects. Furthermore, we demonstrate that the phenotypes caused by the knockdown of Pax1b in zebrafish could not be phenocopied by pax9 MO and could not be rescued by either Pax1a or Pax9 overexpression. We further find that Pax1b affects the expression of col2a1, Uncx4.1, Noggin3, and aggrecan, confirming the role of Pax1b in chondrocyte differentiation and bone maturation. Moreover, we identify an interaction between PAX1 and FOXO1 and find that the interaction was enhanced under hypoxia stress. Together, this evidence for cell death caused by pax1b knockdown provides new insight into the role of the Pax protein family in cell fate determination and tissue specification.