Genome-wide CRISPR Screens Reveal Host Factors Critical for SARS-CoV-2 Infection

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Bone marrow-derived mesenchymal stem cells repair severe acute pancreatitis by secreting miR-181a-5p to target PTEN/Akt/TGF-β1 signaling

BackgroundSevere acute pancreatitis (SAP) is associated with high morbidity and mortality. Bone marrow mesenchymal stem cells (BMSCs) have shown obvious protective effect on SAP. However, little is known about the underlying mechanism. The objective of this study is to unravel the role and regulatory mechanism of miR-181a-5p in BMSCs-mediated pancreatic repair.MethodsBMSCs were isolated from Sprague-Dawley rats and characterized by flow cytometry and Oil Red O staining. Sodium taurocholate- and caerulein-induced models were used as SAP models in vivo and in vitro, respectively. Pancreatic injury were evaluated by H&E and histopathological analysis, as well as by measuring levels of amylase, lipase and cytokines. qRT-PCR and western blotting were performed to detect the level of miR-181a-5p and the protein levels of PTEN/Akt, respectively. ELISA was conducted to detect the levels of TNF-α, IL-1β, IL-6, angiopoietin, IL-4, IL-10 and TGF-β1. The apoptotic rate of AR42 J cells was quantitated by concurrent staining with Annexin-V-FITC and PI.ResultsBMSCs significantly attenuated pancreatic injury in SAP rats by reducing inflammatory infiltration and necrosis, and this effect was abolished by CXCR4 agonist AMD3100. ADM3100 exhibited more severe pancreatic injury and decreased miR-181a-5p levels in the pancreas and serum compared to SAP group. Overexpression of miR-181a-5p in BMSCs (BMSCs-miR-181a-5p) markedly potentiated the protective effect of BMSCs by reducing histological damage and levels of amylase and lipase. Moreover, BMSCs-miR-181a-5p dramatically reduced levels of angiopoietin, TNF-α, IL-1β and IL-6, but induced the levels of IL-4 and IL-10. In caerulein-treated AR42 J cells, co-culturing of BMSCs-miR-181a-5p alleviated caerulein-induced increase of amylase and lipase, and apoptosis via PTEN/Akt/TGF-β1 signaling.ConclusionBMSCs alleviate SAP and reduce inflammatory responses and apoptosis by secreting miR-181a-5p to target PTEN/Akt/TGF-β1 signaling. Hence, BMSCs-miR-181a-5p could serve as potential therapeutic target for SAP.     

利用TMV—U1作为载体表达鼠肝炎病毒S糖蛋白片段

为了验证转基因烟草中表达的外壳蛋白（ＣＰ）能够重新包被侵入的烟草花叶病毒（ＴＭＶ）的假设,利用抗原表位标记的方法观察ＣＰ亚单位在病毒５′端的交换。通过ＰＣＲ 方法将来源于鼠肝炎病毒（ＭＨＶ） Ｓ蛋白的两个小肽段（１１ ａ．ａ．和１５ ａ．ａ．）的ＤＮＡ序列分别插入ＴＭＶ－Ｕ１ ＣＰ基因邻近３′端的两个位点,并构建了带有外源序列的ＴＭＶ 侵染克隆Ｖ９ （１１ ａ．ａ．）和Ｅ１５ （１５ ａ．ａ．）。通过体外转录反应,得到Ｖ９ ＲＮＡ 及Ｅ１５ ＲＮＡ。突变病毒ＲＮＡ 侵染烟草（Ｎｉｃｏｔｉａｎａ ｔａｂａｃｕｍ ）后表现不同特性。Ｖ９ 和Ｅ１５ 侵染ＸａｎｔｈｉＮＮ烟草后同野生型ＴＭＶ一样产生枯斑。但是,当它们侵染Ｘａｎｔｈｉｎｎ 烟草时,Ｖ９ 产生同侵染ＸａｎｔｈｉＮＮ 烟草相同的枯斑,而Ｅ１５的特性同ＴＭＶ－Ｕ１几乎完全相同,能对Ｘａｎｔｈｉｎｎ 烟草进行系统侵染并在叶片中聚集大量的带有外源片段的外壳蛋白,而且病毒的结构极其稳定。Ｖ９ 和Ｅ１５ 特性的差异可能是由于外源片段在外壳蛋白中存在位置的不同影响了外壳蛋白的结构所致     

HBIG,无环鸟苷,干扰素联合对慢性乙型肝炎抗病毒效应观察

本文报道血清ＨＢＶ复制标志阳性的慢乙肝５４例,随机分为治疗组及对照组各２７例进行ＨＢＩＧ、无环鸟苷、干扰素联合近、远期抗病毒效应观察。治疗组为无环鸟苷第一周按２５～２０ｍｇ／ｋｇ／ｄ计后改１７～１５ｍｇ／ｋｇ／ｄ×５３天,共６０天;人白细胞干扰素１×１０６Ｕ肌注每周３次×４周,后改１．０×１０６Ｕ肌注每周２次×６周,共１０周;ＨＢＩＧ４００Ｕ肌注隔日１次,共１０周,对照组仅给予一般“保肝”药物。其中治疗组１８例,对照组１９例进行治后半年到２年追踪观察,结果近、远期ＨＢｃＡｇ、ＤＮＡＰ、ＨＢＶ－ＤＮＡ阴转率治疗组均高于对照组,其中治疗组近、远期ＨＢｃＡｇ,ＨＢＶ－ＤＮＡ阴转率均达４０％以上,明显高于对照组（Ｐ＜０．０５～０．０１）,治疗组近、远期各有４例及２例ＨＢｓＡｇ阴转,而对照组则无一例阴转,从近、远期综合抗病毒效应观察,治疗组全阴率分别为３３．３％、４４．４％,而对照组分别为３．７９％及０％,Ｐ＜０．０１,治疗组无明显毒副反应。对比单用无环鸟苷,全阴率３１．８％;无环鸟苷加干扰素两药联合全阴率３７．５％,均有所提高,达到４４．４％,值得进一步研究。     

Induction of apoptosis and c-myc in L1210 lymphocytic leukemia cells by adenosine

High concentrations of adenosine (Ado), when added to L1210 lymphocytic leukemia cells, resulted in apoptosis or programmed cell death. The apoptotic process was accompanied by distinct morphological changes including chromatin condensation and blebbing of plasma membranes. Extensive DNA fragmentation was correlated with Ado concentrations. Furthermore, apoptosis in these cells was preceded by an early but transient expression of c-myc proto-oncogene, and was not influenced by homocysteine thiolactone added to the cells. Since severe combined immunodeficiency (SCID) is associated with a deficiency of adenosine deaminase, leading to defects in both cellular and humoral immunity, Ado-induced apoptosis may thus be a contributing factor in the pathology of SCID.     

Factores de riesgo asociados a fallecimiento por la variante ómicron de COVID-19: análisis retrospectivo con personas mayores de Canarias

Background and aimsSince the beginning of the COVID-19 pandemic, the elderly population has had the highest rates of complications and mortality. This study aimed to determine the influence of different risk factors on deaths due to the Omicron variant in the Canary Islands.Materials and methodsA retrospective observational study of 16,998 cases of COVID-19 over 40 years of age was conducted in the Canary Islands between August 1, 2022, and January 31, 2023. We extracted sociodemographic data (age and sex) and clinical data (death, vaccination history, hospital admission, previous diseases, and treatments).ResultsAmong the deaths, there was a higher proportion of males aged over 70 years, with diabetes, cardiovascular, renal, respiratory, and systemic diseases, and nursing home residents. Significant differences were observed in the number of doses of the vaccine. The multiple regression model showed that male sex (OR [95% CI] = 1.92 [1.42–2.58]), age (70–79 years, 9.11 [4.27–19.43]; 80–89 years, 21.72 [10.40–45.36]; 90–99 years, 66.24 [31.03–141.38]; 100 years or older, 69.22 [12.97–369.33]), being unvaccinated (6.96, [4.01–12.08]), or having the last dose administered at least 12 months before the diagnosis (2.38, [1.48–3.81]) were significantly associated with mortality.ConclusionsMultiple factors may increase the risk of mortality due to COVID-19 in the elderly population. In our study, we found that only three predictors can effectively explain the variability: older age, male sex, and not being vaccinated or last vaccination date prior to one year.     

Monitoring of early events of experimental woodchuck hepatitis infection: Studies of peripheral blood mononuclear cells by cytofluorometry and PCR

Abstract The peripheral blood mononuclear cells (PBMC) of woodchucks experimentally infected by woodchuck hepatitis virus (WHV) were examined simultaneously for the presence of membrane associated WHV antigens by cytofluorometry, and for WHV DNA and RNA sequences by the polymerase chain reaction (PCR). Four woodchucks were inoculated: two with a well-defined infectious inoculum and two with an inoculum obtained from an animal at the late incubation phase, which was positive for WHV DNA by PCR but still devoid of WHV markers. Infection was demonstrated in all four inoculated woodchucks by the appearance at different times of WHV DNA and WHV antigens in both leucocytes and serum. WHV DNA was first detected by PCR either in the serum (two cases) or in leucocytes (two cases). The mean percentage of cells positive for membrane associated WHsAg or WHcAg detected by cytofluorometry were 37%±25 and 17%±15 respectively. After 8 weeks, all inoculated animals were WHsAg positive in serum. These data suggest that PBMC are involved in the early events of hepadnavirus infection. They also show that sera which are positive by PCR for WHV DNA may transmit viral infection even while still seronegative for WHV markers and for WHV DNA by dot blot.     

重症肺炎患者病原菌分布、临床结局分析及血清尿素氮与肌酐比值、尿素氮与白蛋白比值联合检测的临床意义

摘要 目的：分析重症肺炎（SP）患者病原菌分布和临床结局并探讨血清尿素氮与肌酐比值（UCR）、尿素氮与白蛋白比值（UAR）联合检测的临床意义。方法：选取2019年8月～2022年8月三六三医院收治的107例SP患者,根据临床结局分为死亡组和存活组。分析SP患者病原菌分布情况。采用单因素和多因素Logistic回归分析SP患者临床结局的影响因素,采用受试者工作特征（ROC）曲线分析血清UCR、UAR水平对SP患者临床结局的评估价值。结果：107例SP患者痰液标本中培养出122株病原菌,其中革兰阴性菌75株（61.48%）,革兰阳性菌39株（31.97%）,真菌8株（6.56%）。107例SP患者院内死亡率为40.19%（43/107）。多因素Logistic回归分析显示,年龄增加、肺外并发症≥2个和UCR、UAR升高为SP患者临床结局不良的独立危险因素,氧合指数增加为其独立保护因素（P＜0.05）。ROC曲线分析显示,血清UCR、UAR联合评估SP患者临床结局的曲线下面积（AUC）大于各指标单独评估。结论：SP患者病原菌分布以革兰阴性菌为主,血清UCR、UAR升高为SP患者临床结局不良的独立危险因素,可能成为SP患者临床结局的辅助评估指标,且二者联合评估SP患者临床结局的价值较高。     

多药耐药菌感染重症肺炎患者预后的危险因素分析

摘要 目的：探讨多药耐药菌感染重症肺炎患者预后的危险因素。方法：选取本院2019年5月至2022年5月收治的198例重症肺炎患者,根据患者在ICU住院期间是否死亡分为存活组（121例）和死亡组（77例）。对重症肺炎患者多药耐药菌感染情况,多药耐药G⁺耐药情况,多药耐药G^-耐药情况进行分析,对影响多药耐药菌感染重症肺炎患者预后危险因素的单因素分析,将单因素分析中差异有统计学意义的变量进行多因素Logistic回归分析,筛选影响多药耐药菌感染重症肺炎患者预后的危险因素。结果：198例重症肺炎患者中,多药耐药菌感染患者60例,占比30.30 %,共分离出病原菌290株,其中多药耐药菌65株,占比22.41 %,其中占比比较高的有鲍曼不动杆菌（23.08 %）、铜绿假单胞菌（20.00 %）、金黄色葡萄球菌（20.00 %）、肠炎克雷伯菌（10.77 %）;重症肺炎患者多药耐药G⁺对青霉素、克林霉素、红霉素等具有较高的耐药性,而对万古霉素、替考拉宁、替加环素较为敏感;重症肺炎患者多重耐药G^-对多种抗菌药物均表现出耐药性,其中对头孢他啶、头孢吡肟等具有较高的耐药性;单因素分析结果显示,死亡组患者中男性、年龄≥70岁、APACHEⅡ评分≥26分、有创通气的患者占比显著高于存活组,碳青霉烯类抗生素使用的患者占比显著低于存活组（均P<0.05）,两组患者肺部基础疾病、脑血管疾病、高血压、联合使用其他抗生素的占比,以及两组患者机械通气时间比较无差异（均P>0.05）;纳入多因素非条件Logistic回归模型分析显示,男性、年龄≥70岁、APACHEⅡ评分≥26分、有创通气为多药耐药菌感染重症肺炎患者预后的危险因素（OR=1.568、1.203、2.812、1.674,均P<0.05）,而碳青霉烯类抗生素使用是多药耐药菌感染重症肺炎患者预后的保护因素（OR=0.542,P<0.05）。结论：多药耐药菌感染重症肺炎患者的主要菌株为鲍曼不动杆菌,且男性、年龄≥70岁、APACHEⅡ评分≥26分、有创通气为多药耐药菌感染重症肺炎患者预后的危险因素,而碳青霉烯类抗生素使用是多药耐药菌感染重症肺炎患者预后的保护因素。