苹果黑腐皮壳菌CAP超家族蛋白基因鉴定及毒性功能分析

【目的】CAP(Cysteine-rich secretory protein,Antigen 5 and Pathogenesis related protein 1)超家族蛋白广泛存在于真菌、细菌、动植物等物种,并且参与病原菌的致病过程。本研究旨在鉴定苹果树腐烂病致病菌——苹果黑腐皮壳菌(Valsa mali)的CAP超家族基因并明确CAP超家族基因在病菌致病方面的作用。【方法】通过BLASTP在苹果黑腐皮壳菌全基因组中检索具有CAP保守结构域的基因;利用特异性引物对鉴定到的CAP基因进行PCR扩增和凝胶电泳检测;使用生物信息学软件和在线数据库进行蛋白序列特征和系统发育分析;利用RT-qPCR分析基因在病菌侵染过程中的表达模式;使用特异性引物扩增CAP基因上、下游片段并从PDL2质粒上扩增筛选标记基因;利用Double-joint PCR构建基因敲除盒并通过PEG介导的原生质转化技术进行基因敲除和基因回补;以遗传霉素(G418)抗性为筛选标记并利用4对引物PCR检测获得基因敲除突变体;以潮霉素(HPH)抗性为筛选标记获得基因回补菌株;通过对菌株进行培养皿内生长试验明确基因对病菌营养生长的影响;通过离体苹果枝条接种试验分析该病菌CAP超家族基因的毒性功能。【结果】在苹果黑腐皮壳菌中鉴定到3个具有CAP保守结构域的基因,分别命名为VmPR1a、VmPR1b和VmPR1c。序列特征分析发现,3个CAP蛋白均包含4个保守区:N端信号肽、N端延伸区(NTE)、CAP保守功能域和C端延伸区(CTE)。系统发育分析显示,3个CAP蛋白聚于不同的进化支,VmPR1a聚于clade2进化支并与粗糙链孢霉(Neurospora crassa)CAP蛋白进化关系较近;VmPR1b聚于clade3且与镰孢菌属(Fusarium spp.)CAP蛋白的进化关系更近;VmPR1c聚于clade1,并且也与粗糙链孢霉的CAP蛋白进化关系较近。RT-qPCR分析结果显示,VmPR1a、VmPR1b和VmPR1c在病菌侵染早期(6 h和12 h)均显著上调表达。利用PEG遗传转化技术获得VmPR1a、VmPR1b和VmPR1c敲除突变体(ΔVmPR1a-7/23、ΔVmPR1b-20/31和ΔVmPR1c-26/40);营养生长观察发现,所有敲除突变体生长表型与野生型菌株03-8均无明显差异;致病力检测发现,ΔVmPR1b-20/31致病力较野生型无明显变化,而ΔVmPR1a-7/23和ΔVmPR1c-26/40致病力较野生型显著下降。将VmPR1a和VmPR1c分别回补至ΔVmPR1a和ΔVmPR1c,回补菌株(VmPR1a/C和VmPR1c/C)致病力恢复至野生型水平。【结论】苹果黑腐皮壳菌中存在3个CAP超家族基因(VmPR1a、VmPR1b和VmPR1c),其中VmPR1a和VmPR1c是苹果黑腐皮壳菌重要的毒性因子。

组合型人工肝技术治疗中毒性肝功能衰竭的临床研究

探究组合型人工肝技术治疗中毒性肝功能衰竭的临床价值。[研究时间段]:2020年8月~2021年3月,[研究对象]:70例因中毒性肝功能衰竭疾病而在本院接受治疗的患者,随机分组,两组各有35例患者,对照组实施单纯血浆置换治疗(PE),实验组实施组合型人工肝技术治疗(HBL),探究两组患者生物化学指标(总胆红素、血氨、白蛋白、胆碱酯酶、凝血酶原活性)。结果:对中毒性肝功能衰竭患者实施组合型人工肝技术治疗后,有效改善了实验组总胆红素、血氨、白蛋白、胆碱酯酶、凝血酶原活性等生物化学指标,两组间数据对比,差异明显,(P<0.05)。结论:实施组合型人工肝技术治疗后,有效改善了中毒性肝功能衰竭患者的临床数据以及肝功能指标,提升患者的预后水平,有较好的临床推广价值。

Toll样受体3在脓毒性心功能障碍中作用的研究进展

Toll样受体3（Toll—like Receptor 3，TLR3）能特异性识别外源性病毒双链RNA及Poly：C，在抗病毒免疫反应中发挥重要作用。更为重要的是，最近的研究发现，它也可以识别宿主凋亡及坏死细胞所释放的细胞外RNA（extracellular RNA，exRNA），在细菌性脓毒血症诱导的心功能障碍中发挥核心作用。本文就TLR3在脓毒血症心功能障碍中的作用研究进展进行综述。

超声心动图联合B型脑钠肽在脓毒性心肌功能障碍患者预后评估中的价值

目的探讨超声心动图联合B型脑钠肽(BNP)在脓毒性心肌功能障碍(SIMD)患者预后评估中的价值。方法选取2020年1月至2022年1月就诊于新疆维吾尔自治区人民医院的脓毒症/脓毒性休克患者82例。根据后期剔除、完成随访、超声心动图检查结果及死亡情况分组。比较不同组间相关血清学指标及超声心动图指标,采用多因素Logistic回归方法分析影响患者心肌功能障碍发生的相关因素,采用受试者工作特征(ROC)曲线评估超声心动图联合BNP对SIMD患者预后的早期预测价值。结果观察过程中2例分别因心肺复苏、急性冠状动脉综合征剔除,2例入住重症监护病房72 h内自动出院,最终入组并完成随访患者78例。根据患者是否存在左心室舒张功能障碍,分为心肌功能障碍组(32例)和心肌功能正常组(46例);根据入院28 d死亡情况分为死亡组(28例)和存活组(50例)。心肌功能障碍组和死亡组血清BNP、白细胞介素6、心肌肌钙蛋白T、肌酸激酶、肌酸激酶同工酶、左心室舒张末期内径、左心室收缩末期内径、二尖瓣舒张早期血流峰值速度(E)/二尖瓣瓣环舒张早期血流峰值速度(e′)比值分别高于心肌功能正常组和存活组,E/二尖瓣舒张晚期血流峰值速度(A)比值分别低于心肌功能正常组和存活组,差异均有统计学意义(均P<0.05)。Logistic回归分析结果显示,BNP(比值比=1.947)、心肌肌钙蛋白T(比值比=1.615)、E/A比值(比值比=1.785)及E/e′比值(比值比=1.162)是脓毒症患者发生心肌功能障碍的独立影响因素(均P<0.05)。超声心动图联合BNP对SIMD患者预后预测的ROC曲线下面积为0.906,敏感度为90.6%,特异度为90.8%。结论BNP水平和超声心动图指标作为早期监测手段,在一定程度上能够反映SIMD的程度,并对SIMD患者预后判断具有重要意义。

不同监测手段对脓毒性休克并发心功能不全患者治疗效果的影响

目的比较脉搏指示连续心排血量(PICCO)和中心静脉压(CVP)监测对脓毒性休克并发心功能不全患者治疗的指导意义。方法选取于2017年6月至2019年12月在连州市人民医院就诊的60例脓毒性休克并发心功能不全患者,采用随机数字表法分为常规组和观察组各30例。常规组进行CVP监测指导治疗,观察组进行PICCO监测指导治疗。比较两组治疗效果及血流动力学指标。结果两组采用不同监测手段实施治疗方案后B型脑钠肽(BNP)、心率(HR)、CVP均降低,而平均动脉压(MAP)升高,两组BNP、HR、MAP比较,差异有统计学意义(P<0.01)。两组治疗后随着时间急性生理学与慢性健康状况评分系统Ⅱ(APACHEⅡ)评分均不同程度降低,与常规组,观察组治疗后降低更显著,差异有统计学意义(P<0.01)。与治疗前相比,观察组体循环阻力指数(SVRI)、心排血指数(CI)均显著升高,CO则显著降低,差异有统计学意义(P<0.01)。结论较常规监测方式,PICCO监测指标指导患者早期容量复苏方案效果更佳,有助于病情控制。

中毒性急性肾功能衰竭58例临床分析

中毒性急性肾功能衰竭是临床常见的危重病,我院1988年5月至1998年5月抢救中毒性急性肾功衰58例,现将临床资料分析如下.

Toll样受体3在脓毒症并心功能障碍患儿中的表达及意义

目的 探讨脓毒症并心功能障碍患儿外周血Toll样受体3（TLR3）的表达及其临床意义.方法 采用前瞻性对照研究方法,选取2015年10月至2017年3月在汕头市中心医院儿科住院且诊断符合脓毒血症不伴心功能障碍的患儿20例（脓毒血症组）、脓毒血症并心功能障碍患儿24例（脓毒性心功能障碍组）、非脓毒性心功能障碍的患儿10例（非脓毒性心功能障碍组）和健康体检儿童30例（对照组）作为研究对象,各组儿童均在入院24 h内留取新鲜外周血2 ml,用流式细胞仪检测单核细胞内TLR3的表达.结果 脓毒血症组TLR3[（55.1±30.2）％]、脓毒性心功能障碍组TLR3[（51.9±33.5）％]、非脓毒性心功能障碍组TLR3[（61.3±26.7）％]表达均较正常对照组儿童TLR3[（5.7±2.4）％]显著升高,差异均有统计学意义（均P＜ 0.01）;但3个病例组（脓毒血症组、脓毒性心功能障碍组和非脓毒性心功能障碍组）间TLR3水平比较,差异均无统计学意义（均P＞0.05）.结论 TLR3信号通路的激活可能是脓毒血症及心功能障碍的重要机制,抑制TLR3的过度表达可能成为减轻脓毒血症和心功能障碍潜在的治疗靶点.

中毒性肝损害研究现状

西药、中药、矿物药所致中毒性肝损害临床上并不少见，但医务人员尤其患者并不高度关注这方面的病因。为此，作者通过阅读相关文献，就药物性肝损害的分类、发病机理、诊断及其防治进行扼要综述。

儿童脓毒性休克中的心肌功能障碍——值得关注的问题

【摘要】儿童脓毒性心功能障碍（脓毒性心肌病）在严重脓毒症及脓毒性休克患儿中十分常见，其临床诊断主要靠心脏超声结果。本文回顾了脓毒性心肌病的诊断及预后，并简要回顾了脓毒性心肌病潜在机制的研究热点及争议之处。

Hepatotoxicity of NONI juice: Report of two cases

NONI juice (Morinda citrifolia) is an increasingly popular wellness drink claimed to be beneficial for many illnesses.No overt toxicity has been reported to date. We present two cases of novel hepatotoxicity of NONI juice. Causality of liver injury by NONI juice was asses-sed. Routine laboratory tests and transjugular or percutaneous liver biopsy were performed. The first patient underwent successful liver transplantation while the second patient recovered spontaneously after cessation of NONI juice.A 29-year-old man with previous toxic hepatitis associated with small doses of paracetamol developed sub-acute hepatic failure following consumption of 1.5 L NONI juice over 3 wk necessitating urgent liver transplantation. A 62-year-old woman without evidence of previous liver disease developed an episode of self-limited acutehepatitis following consumption of 2 L NONI juice for over 3 mo. The most likely hepatotoxic components of Morinda citrifolia were anthraquinones. Physicians should be aware of potential hepatotoxicity of NONI juice.

Ceftriaxone-induced toxic hepatitis

Toxic hepatitis or drug-induced liver injury encompasses a spectrum of clinical disease ranging from mild biochemical abnormalities to acute liver failure.The advantages of a long half-life,wide spectrum,high tissue penetration rate,and a good safety profile,make ceftriaxone,a third-generation cephalosporin,a frequent choice in the treatment of childhood infections.Previous studies have reported a few cases of high aspartate aminotransferase and alanine aminotransferase levels,along with three cases of hepatitis caused by ceftriaxone.Here,we report a case of drug-induced toxic hepatitis in a patient who was treated with ceftriaxone for acute tonsillitis.

Analyses of prognostic indices of chronic liver failure caused by hepatitis virus

AIM: To analyze the related indices about the prognoses of chronic liver failure caused by hepatitis virus. METHODS: Retrospectively reviewed 320 cases of chronic liver failure caused by hepatitis viruses. An improved group and an ineffective group (IG) were made to compare and analyze their clinical manifestations, laboratory examination indices and complications. Logistic regression was also carried out. RESULTS: There were significant differences (P<0.05) between the improved group and the IG upon such indices as age, bilirubin, prothrombin time, albumin, alpha fetoprotein, the size of liver and complications (P<0.05). The regression formula was as follows: P=1/(1+e^(-y)) (y=1.7262-0.0948X_1+2.9846X_2+0.6992X_3+1.6019X_4+2.0398X_5). (Note: X_1-Prothrombin activity; X_2-digestive tract hemorrhage; X_3-hepatic encephalopathy; A_4-hepatorenal syndrome; X_5-pulmonary infection.). CONCLUSION: Laboratory examination such as bilirubin, prothrombin time and alpha fetoprotein can be regarded as indices of the prognoses of chronic liver failure caused by hepatitis. Moreover, the regression equation can evaluate prognoses more comprehensively and direct our treatments.

Evaluation of a novel hybrid bioartificial liver based on a multi-layer flat-plate bioreactor

AIM: To evaluate the efficacy and safety of a hybrid bioartificial liver (HBAL) system in the treatment of acute liver failure. METHODS: Canine models with acute liver failure were introduced with intravenous administration of D-galactosamine. The animals were divided into: the HBAL treatment group (n = 8), in which the canines received a 3-h treatment of HBAL; the bioartificial liver (BAL) treatment group (n = 8), in which the canines received a 3-h treatment of BAL; the non-bioartificial liver (NBAL) treatment group (n = 8), in which the canines received a 3-h treatment of NBAL; the control group (n = 8), in which the canines received no additional treatment. Biochemical parameters and survival time were determined. Levels of xenoantibodies, RNA of porcine endogenous retrovirus (PERV) and reverse transcriptase (RT) activity in the plasma were detected. RESULTS: Biochemical parameters were significantly decreased in all treatment groups. The TBIL level in the HBAL group was lower than that in other groups (2.19 ± 0.55 mmol/L vs 24.2 ± 6.45 mmol/L, 12.47 ± 3.62 mmol/L, 3.77 ± 1.83 mmol/L, P < 0.05). The prothrombin time (PT) in the BAL and HBAL groups was significantly shorter than the NBAL and control groups (18.47 ± 4.41 s, 15.5 ± 1.56 s vs 28.67 ± 5.71 s, 21.71 ± 3.4 s, P < 0.05), and the PT in the HBAL group was shortest of all the groups. The albumin in the BAL and HBAL groups significantly increased and a significantly higher level was observed in the HBAL group compared with the BAL group (27.7 ± 1.7 g/L vs 25.24 ± 1.93 g/L). In the HBAL group, the ammonia levels significantly decreased from 54.37 ± 6.86 to 37.75 ± 6.09 after treatment (P < 0.05); there were significant difference in ammonia levels between other the groups (P < 0.05). The levels of antibodies were similar before and after treatment. The PERV RNA and the RT activity in the canine plasma were all negative. CONCLUSION: The HBAL showed great efficiency and safety in the treatment of acute liver failure.

Epidemiological and pathogenic study on the outbreak of toxic shock syndrome and meningocephalitis caused by swine streptococcus

Objective: To reveal the relationship between the biological characteristics of pathogen and the pig to human spread of the epidemic and infectious disease in 1998 in East China. Methods: Epidemiological survey, pathological examination of pigs and patients, and pathogen isolation were performed. Results: The disease had a character of quick onset, serious symptoms, short course and high mortality. The clinical manifestations and pathological changes of the disease were high fever, sometimes with nausea, vomiting and diarrhea, then might develop to myositis, fascitis, DIC, multiple organ failure, shock and usually died in 2 3 d. Among 25 patients, 16 manifested clinically as streptococcal toxic shock syndromes and 9 streptococcal meningiocephalitis syndrome. The mortality was 81 25% and 11 11% respectively. Pathogenic bacteria isolated from diseased pigs and patients were found to have some common characteristics in morphology, staining and biological characters. Conclusion: The pathogen isolated from the blood of patients and pigs were identified as streptococci.

Hemorrhagic Fever with Renal Syndrome Complicated by Orchitis

Hemorrhagic fever with renal syndrome (HFRS) is a disease caused by viruses of the family Bunyaviridae,genus Hantavirus.HFRS from Dobrava virus (DOBV) is a seldom reported disease in Albania.Clinically HFRS is manifested as mild,moderate,or severe.Therefore,the number of cases of Hantavirus' infection may be underestimated,and should be included in the differential diagnosis of many acute infections,hematologic diseases,acute abdominal diseases and renal diseases complicated by acute renal failure.We report here an atypical presentation of HFRS from Dobrava virus complicated by orchitis with a positive outcome.

脓毒性心肌病临床诊治进展

自1984年提出脓毒性心肌病(SICM)的概念以来,目前仍然没有SICM统一的诊断、治疗标准。近20年有越来越多关于SICM的发病机制、诊断以及管理策略的研究,认识到SICM可能诊断不足,并且与预后不良相关。目前对于SICM没有基于高质量临床证据的管理建议,SICM的治疗是以脓毒性休克的治疗为基础,以血流动力学和组织灌注为导向的支持性措施。连续动态床旁超声心动图有利于早期识别SICM,有助于实施个体化治疗和管理,以改善预后。

Regulatory effects of coenzyme Q10 on the immunological activity of mice

To examine how coenzyme Q10（CoQ10） regulates immunity, experiments using low, middle and high doses of CoQ10 were conducted in mice to confirm its non-toxicity and non-genotoxicity. Delayed type hypersensitivity（DTH） and MTT assays were used to to examine various lymphocyte transformations, the proliferation of antibody-producing cells, the phagocytotsis activity of macrophages, and the activity of nature killer cell（NK）. High-dose（0.50 g/kg.bw） CoQ10 increased DTH levels and promoted the proliferation of antibody-producing cells and levels of red blood cell hemolysis. Medium and high doses enhanced the phagocytic ratio of macrophages but did not influence other indexes. These results showed that the applied CoQ10 did not exhibit any toxicity or genotoxicity, and CoQ10 can actually improve immunologic function in mice.