慢加急性肝衰竭不同预后患者血浆外泌体差异蛋白的生物信息学分析

目的筛选不同预后慢加急性肝衰竭(ACLF)患者血浆外泌体中的差异蛋白,分析其功能及生物学过程,为患者临床诊断提供参考依据。方法前瞻性选取2019年7月—10月于首都医科大学附属北京佑安医院住院确诊的ACLF患者10例,随访90 d,患者死亡或肝移植归入肝移植/死亡组(n=5),患者存活归入生存组(n=5),两组一般资料指标比较采用Mann-Whitney U秩和检验。采用非标记(Label-free)定量蛋白质组学技术对血浆外泌体蛋白进行鉴定和定量分析,筛选差异蛋白并进行功能富集分析,使用R-3.5.1软件对差异蛋白进行层次聚类分析,分析其参与的生物学过程。结果外泌体蛋白质组学分析共鉴定出860种蛋白,以倍数上调>1.2倍或下调>1.2倍且P<0.05的标准筛选出差异表达蛋白116种,与肝移植/死亡组相比,生存组上调蛋白62种、下调蛋白54种。生物信息学分析结果显示,这些蛋白主要参与了免疫反应、信号转导、囊泡介导的转运、细胞死亡和增殖等生物学过程,并与炎症反应、糖类和氨基酸代谢、肝细胞损伤及再生等信号通路密切相关。结论非标记定量蛋白质组学技术筛选出的差异蛋白可能作为ACLF早期诊断及预后判断的血清学标志物。     

新型冠状病毒肺炎患者40例临床特点与舌象关系研究

目的研究新型冠状病毒肺炎(COVID-19)患者的临床特点与舌象表现的关系。方法观察40例COVID-19患者的症状、体征、实验室检查、影像学检查结果,并结合舌象进行分析。结果(1)舌象表现:40例患者中,舌质红者40例(100%),其中舌质暗红者19例(47.5%);少苔者5例(12.5%),苔白腻者27例(67.5%),苔黄腻者8例(20.0%)。(2)临床表现:40例患者中,发热27例(67.5%),其中低热(37.3~37.9℃)18例(45.0%)、中热(38~38.9℃)8例(20.0%)、高热(≥39℃)1例(2.5%);咳嗽25例(62.5%),乏力13例(32.5%),咽痛6例(15.0%),肌肉酸痛12例(30.0%),腹泻2例(5.0%)。舌红少苔者症状多轻微,仅有咳嗽、咽痛、乏力等症状,咽痛症状较舌苔厚腻者更为多见,差异有统计学意义(P<0.05);舌苔白腻、黄腻者则以咳嗽、发热的特征性临床表现为主,其中舌苔白腻者以低热为主,舌苔黄腻者发热多为38℃以上的中热或高热,差异有统计学意义(P<0.05)。(3)影像学表现:胸部X线片提示,40例患者中双侧肺部炎症患者13例(32.5%);单侧肺部炎症患者18例(45.0%),其中左肺炎症患者7例(17.5%),右肺炎症患者11例(27.5%);其余9例(22.5%)患者胸部X线片检查均未见明显异常。舌红少苔者影像学检查多无异常,胸部X片提示肺部炎症多为舌苔厚腻者,其中双肺炎症多见于舌红苔黄腻者,差异有统计学意义(P<0.05)(4)实验室检查:40例患者血白细胞计数为(5.17±1.55)×10^9/L,其中异常减低患者3例(7.5%);淋巴细胞计数为(1.39±0.75)×10^9/L,其中异常减低患者8例(20.0%),随着舌苔逐渐厚腻的变化,血白细胞计数逐渐升高,淋巴细胞计数逐渐下降,差异有统计学意义(P<0.05)。结论COVID-19患者舌红少苔多见于疾病早期病程较短者(1~2d),症状多轻微;伴随病程进展(病程≥3d),部分患者舌苔逐渐增多,表现为厚腻苔,症状以咳嗽、发热的特征性临床表现为主,反映了感染逐渐进展的情况,由此及时总结出COVID-19宜采用清热宣肺利湿、不可过用苦寒的治疗原则。     

PLCε基因敲除小鼠移植性肝癌模型的建立与分析

目的利用PLCε基因敲除小鼠建立移植性肝癌动物模型,以探讨PLCε基因在肝肿瘤生长发育过程中的作用。方法选取PLCε+/+小鼠15只为对照组,选取PLCε+/+(野生型)和PLCε-/-(敲基因型)小鼠各15只为模型组I和II,沿腹中线开腹后将H22细胞接种到模型组小鼠肝脏实质内,对照组注射生理盐水。于注射后第15天行剖腹探查,观察各组成瘤率及肿瘤体积,并进行肿瘤病理学分析。结果各组小鼠的存活率均为100%。模型组I肿瘤移植成功率为100%,肿瘤体积平均为(65.21±5.25)mm3。模型组II的肿瘤移植成功率为53.3%,肿瘤体积平均为(23.46±3.47)mm3。模型组I的肿瘤平均体积明显大于模型组II(P0.05)。模型I组和II组病理检查均证实为原位肝细胞瘤。结论成功建立了PLCε敲基因小鼠移植性肝癌动物模型,验证了PLCε敲基因小鼠具有抑制肝肿瘤生长的特性。     

Hypodermin C improves the survival of kidney allografts

Although immunosuppressive therapies have made organ transplantation a common medical procedure worldwide, chronic toxicity is a major issue of long-term treatment. One method to improve such therapies is the application of immunomodulatory agents from parasites, such as Hypoderma lineatum (Diptera: Oestridae). Hypodermin C (HC) is an enzyme secreted by H. lineatum larvae, and our previous study showed that recombinant HC could degrade guinea pig C3 and inhibit the complement pathway in vitro, suggesting potential activity for inhibiting transplant rejection. However, such properties have not been fully demonstrated in vivo. In this study, we investigated the impact of HC on a fully MHC-mismatched, life-sustaining, murine model of kidney allograft rejection using B6 donors and BABL/c (HC transgenic or wild-type) recipients. Kidney grafts were analyzed by histology, immunohistochemistry and western blotting. The results suggested that HC could effectively inhibit kidney allograft rejection. These findings suggest HC is a promising strategy to improve the survival of human implants.     

Complement-independent dengue virus type 1 infection-enhancing antibody reduces complement-dependent and -independent neutralizing antibody activity

Dengue fever and dengue hemorrhagic fever are globally important mosquito-transmitted viral diseases. However, the only licensed vaccine is not highly protective. Viremia is related to disease severity in infected humans, and it is thought to be reduced by neutralizing antibodies but increased by infection-enhancing antibodies. We established an assay system to measure the balance between neutralizing and enhancing antibodies and found that most dengue-immune individuals in endemic areas carry complement-independent enhancing antibodies (CiEAb). Studying CiEAb is important for dengue vaccine development because the enhancing activity of CiEAb does not decrease in the presence of complement, which can reduce the enhancing activity of other antibodies in vitro. Here, we investigated the effects of CiEAb on the activity of neutralizing antibodies (mainly, complement-dependent neutralizing antibodies; CdNAb) using cocktails of mouse monoclonal antibodies (MAbs) against dengue virus type 1 (DENV-1). These cocktails included MAbs with enhancing activity only (represented by D1-V-3H12 [3H12]) or neutralizing activity only (represented by D1-IV-7F4 [7F4]). Because 3H12, an IgG1 subclass antibody, is complement-independent and cross-reacted with all dengue serotypes, it is a suitable model of CiEAb. An approximately equal amount of 3H12 abolished the neutralizing activity of 7F4. The complement-dependent neutralizing activities of the IgG2a and IgG2b variants of 7F4 were also completely inhibited by ⩾3-fold concentrations of the IgG1 variant. The complement-dependent antibody activities of other anti-DENV-1 MAbs and those of MAbs directed against other serotypes were inhibited 50% by 3H12 at various mixing ratios, ranging from one-hundredth to 10-fold. The complement-dependent neutralizing activities of dengue-immune mouse ascites fluids were also effectively inhibited by 3H12. This suggests that concomitantly induced CiEAb exerts an unwanted effect on the protective capacity of a vaccine. Thus, the effective inhibition of the neutralizing activity of CdNAb by CiEAb has implications for dengue pathogenesis and vaccine development.     

Drug decriminalization and the price of illicit drugs

BackgroundThis study is an empirical assessment of the impact of the drug decriminalization policy followed by Portugal in July 2001, on the price of illicit drugs.MethodsThe analysis is performed using a difference-in-differences approach and the Synthetic Control Method in order to construct a synthetic control unit from a convex combination of countries.ResultsThe results suggest that the prices of opiates and cocaine in the post-treatment period did not decrease in the sequence of the policy change.ConclusionWe conclude that the drug decriminalization policy seems to have caused no harm through lower illicit drugs prices, which would lead to higher drug usage and dependence.     

CAROTENOID, RETINOID AND VITAMIN E STATUS OF THE OROPHARYNGEAL MUCOSA IN THE ALCOHOLIC

Concentrations of carotenoids, retinoids and tocopherols weredetermined in the homogenate of macroscopically normal appearingoropharyngeal mucosa from 10 chronic alcoholics and from 11control patients. All the alcoholics except one had oropharyngealcancer. No significant difference was found in tissue levelsof carotenoids and tocopherols between alcoholics and controls.Furthermore, in seven of 11 controls, retinol was undetectablein the oropharyngeal mucosa, while in the alcoholics only twoout of 10 had unmeasurable retinol levels. These results donot support the concept that ethanol-associated oropharyngealcarcinogenesis is due, at least in part, to local deficienciesin retinoids, carotenoids or     

Serum Procollagen Type III N-Terminal Peptides and Laminin P1 Peptide in Alcoholic Liver Disease

The appearance of perivenular fibrosis on liver biopsy reflects the beginning of the fibrotic process that ultimately results in liver cirrhosis. To examine whether the fibrogenic activity can be detected by blood tests, we evaluated whole antibody radioimmunoassay (RIA) of procollagen type III N-terminal peptides (P-III-P), RIA of these peptides using Fab fragments (Fab-P-III-P), and RIA of the laminin P1 peptide in alcoholics within 1 week of alcohol abstinence. The Fab-P-III-P levels in subjects with perivenular fibrosis were significantly higher than those in patients with simple fatty liver. Values in 63% of subjects with perivenular fibrosis exceeded the upper limit of the fatty liver group. Patients with simple fatty liver had significantly lower values than nonalcoholic controls. Serum levels of P-III-P and laminin were elevated in patients with alcoholic hepatitis and correlated well with the degree of inflammation. With abstinence, Fab-P-III-P levels increased in all alcoholics. P-III-P values increased in patients with normal P-III-P values on admission. By contrast, the values of laminin decreased during abstinence. Therefore, to interpret serum levels of Fab-P-III-P, P-III-P, and laminin, the duration of abstinence must be taken into consideration. P-III-P, Fab-P-III-P and laminin measurements in the serum within 1 week of abstinence can contribute to the detection of alcoholic liver disease and the determination of its stage.