Effect of Xiaoyu Zhixue Tablet (消瘀止血片) on Expression ofPlatelet Membrane Glycoproteins in Patients withHemorrhagic Thrombopathy

Objective: To observe the effect of Xiaoyu Zhixue tablet (消瘀止血片,XYZXT) on the expression of platelet membrane glycoproteins in patients with hemorrhagic thrombopathy, and to explore its possible mechanism. Methods: The total of 148 patients with hemorrhagic thrombopathy were randomly divided into two groups, the traditional Chinese medicicne (TCM) group (n=98) treated with XYZXT and the Western medicine (WM) group (n=50) treated with adrenosin, vitamins C, K and P, both for 6 months. The therapeutic effect and the recovery rate of platelet aggregation in the two groups were observed. And platelet membrane glycoprotein (GP) Ⅰb/Ⅸ, GPⅡb/Ⅲa complexes, GPⅠb, GPⅡb, GP Ⅲa and P-selectin were analyzed by flow cytometry in both groups before and after treatment and also in 34 normal healthy subjects. Results: The total effective rate of hemostasis was 89. 8% in TCM group and 54. 0% in the WM group (x2=45.83, P<0.01), and the recovery rate of platelet aggregation was 72.4% and 4.0% respectively (x2=62.06, P<0.01). The fluorescence intensity of GP Ⅰ b/Ⅸ, GPⅡb/Ⅲa complexes, GPⅠb, GPⅢa and P-selectin were lower in both groups before treatment than those in the healthy subjects. Expression of above-mentioned marks was elevated in TCM group after 6 months' therapy, which was insignificantly different as compared with the healthy subjects (P>0.05) and higher than those in the WM group (P<0.05). Conclusion: One of the mechanisms in treating hemorrhagic thrombopathy with XYZXT is that it could regulate the expression of GP Ⅰb/Ⅸ, GPⅡ b/Ⅲa complexes, GPⅠb, GPⅢa and P-selectin at the level of receptor protein.

Multi-objective optimization of the cathode catalyst layer micro-composition of polymer electrolyte membrane fuel cells using a multi-scale,two-phase fuel cell model and data-driven surrogates

Polymer electrolyte membrane fuel cells(PEMFCs)are considered a promising alternative to internal combustion engines in the automotive sector.Their commercialization is mainly hindered due to the cost and effectiveness of using platinum(Pt)in them.The cathode catalyst layer(CL)is considered a core component in PEMFCs,and its composition often considerably affects the cell performance(V_(cell))also PEMFC fabrication and production(C_(stack))costs.In this study,a data-driven multi-objective optimization analysis is conducted to effectively evaluate the effects of various cathode CL compositions on Vcelland Cstack.Four essential cathode CL parameters,i.e.,platinum loading(L_(Pt)),weight ratio of ionomer to carbon(wt_(I/C)),weight ratio of Pt to carbon(wt_(Pt/c)),and porosity of cathode CL(ε_(cCL)),are considered as the design variables.The simulation results of a three-dimensional,multi-scale,two-phase comprehensive PEMFC model are used to train and test two famous surrogates:multi-layer perceptron(MLP)and response surface analysis(RSA).Their accuracies are verified using root mean square error and adjusted R^(2).MLP which outperforms RSA in terms of prediction capability is then linked to a multi-objective non-dominated sorting genetic algorithmⅡ.Compared to a typical PEMFC stack,the results of the optimal study show that the single-cell voltage,Vcellis improved by 28 m V for the same stack price and the stack cost evaluated through the U.S department of energy cost model is reduced by$5.86/k W for the same stack performance.

Multi-objective optimization of membrane structures based on Pareto Genetic Algorithm

A multi-objective optimization method based on Pareto Genetic Algorithm is presented for shape design of membrane structures from a structural view point.Several non-dimensional variables are defined as optimization variables,which are decision factors of shapes of membrane structures.Three objectives are proposed including maximization of stiffness,maximum uniformity of stress and minimum reaction under external loads.Pareto Multi-objective Genetic Algorithm is introduced to solve the Pareto solutions.Consequently,the dependence of the optimality upon the optimization variables is derived to provide guidelines on how to determine design parameters.Moreover,several examples illustrate the proposed methods and applications.The study shows that the multi-objective optimization method in this paper is feasible and efficient for membrane structures;the research on Pareto solutions can provide explicit and useful guidelines for shape design of membrane structures.

Expression of Platelet Membrane Glycoprotein Ib/Ⅸ/Ⅴ Complex,a Receptor of Thrombin,in Patients with Hemorrhagic Thrombopathy

To investigate the role of platelet membrane glycoprotein （GP） Ib/Ⅸ/Ⅴ complex and its subunit GP Ibα in patients with hemorrhagic thrombopathy （HT）, the expressions of GP Ib/Ⅸ/Ⅴ complex and GP Ibα,defined as mean fluorescence intensity （MFI）, were assessed by flow cytometry. The maximum aggregation of platelet was determined by turbidity method. These indicators were compared among 68 HT patients with the presenting complaint of hemorrhage, 33 well-controlled HT patients and 32 normal healthy subjects. The results showed that the MFI of GP Ib/Ⅸ /Ⅴ complex and GP Ibα was markedly lower in HT patients with current hemorrhage than that in the healthy subjects, with difference being statistically significant （P〈0.05）. There was no significant difference in the expressions of GP Ib/ Ⅸ/ Ⅴ complex and GP Ibα between well-controlled HT patients and normal healthy subjects （P〉0.05）. It was concluded that the expression of GP Ib/Ⅸ /Ⅴ complex, the receptor of thrombin and von Willebrand factor, was down-regulated in HT patients with current hemorrhage, which might result in the dysfunction of platelet aggregation and recurrence of HT.

Association of the Platelet Membrane Glycoprotein Ⅰa C807T Gene Polymorphism with Aspirin Resistance

To explore the correlation between the C807T polymorphism of platelet membrane glycoprotein Ⅰa （GP Ⅰa） gene and aspirin resistance in Chinese people, 200 patients with high-risk of atherosclerosis took aspirin （100 mg/d） for 7 days. Platelet aggregation function was detected using adenosine diphosphate （ADP） and arachidonic acid （AA） before and after the administration of aspirin. Then the subjects were divided into three groups according to the results of platelet aggregation function： an aspirin resistant （AR） group, an aspirin semi-responder （ASR） group and an aspirin-sensitive （AS） group. Platelet GP Ⅰa gene 807CT polymorphism was examined by means of polymerase chain reaction-sequence specific primers （PCR-SSP）. The results showed that T allelic frequency in AR group and ASR group were higher that of AS group （P〈0.005）, and the prevalence of genotypes （TT＋TC） of these two groups was significantly higher than that in AS group （P〈0.05）. Platelet GP Ⅰa T allele was significantly associated with aspirin resistance as revealed by multiple logistic regression （OR=3.76, 95% CI： 2.87-9.58）. The results suggest that inherited platelet GP Ⅰa variations may have an important impact on aspirin resistance and the presence of GP Ⅰa T allele may be a marker of genetic susceptibility to aspirin resistance.

Identification and analysis based on genetic algorithm for proton exchange membrane fuel cell stack

The temperature of proton exchange membrane fuel cell stack and the stoichiometric oxygen in cathode have relationship with the performance and life span of fuel cells closely. The thermal coefficients were taken as important factors affecting the temperature distribution of fuel cells and components. According to the experimental analysis, when the stoichiometric oxygen in cathode is greater than or equal to 1.8, the stack voltage loss is the least. A novel genetic algorithm was developed to identify and optimize the variables in dynamic thermal model of proton exchange membrane fuel cell stack, making the outputs of temperature model approximate to the actual temperature, and ensuring that the maximal error is less than 1 ℃. At the same time, the optimum region of stoichiometric oxygen is obtained, which is in the range of 1.8-2.2 and accords with the experimental analysis results. The simulation and experimental results show the effectiveness of the proposed algorithm.

Redistribution of Platelet Membrane Glycoprotein IV and Release of Intracellular α-granule Thrombospondin in Patients with Chronic Myelogenous Leukemia

The redistribution of platelet membrane glycoprotein IV (GPIV) and the release of intracellular Q-granule thrombospondin (TSP) were examined and the inhibition of 5-thromboglobulin (&TG) and platelet factor 4 (PF4) in patients with chronic myelogenous leukemia (CML) was observed and quantitation of β-TG and PF4 in sera was conducted. GPIV in inactive platelet from CML was 36080±17010 molecules/platelet as compared with 13190±4810 from the controls (P<0,01), No abnormality was found in the distribution of platelet membrane GPIb and GPIIb/III.(P>0. 05). The GPIV redistribution on active platelet membrane induced thrombin (1U/ml) from CML and healthy donors was 44320132310 and 228001 12700 molecules/platelet respectively (P<0. 01 ). The difference in the release of intracellular Q-granule TSP between CML and the control group was not found (P>0.05). There was no direct correlation between GPIV expression and TSP binding after platelet activation. The high leveIs of β-TG and PF4 in sera inhibited release of intracellular a-granule TSP in vitro. These results indicate that the abnormality of platelet membrane GPIV is a common marker in CML, therefore the specific increase of platelet GPIV in patients with CML may be a useful tool for the diagnosis and monitoring of the platelet dysfunction. The release of interna1 TSP pools is hindered by either β-TG or PF4 in sera.

Membrane glycoprotein M6A promotes μ-opioid receptor endocytosis and facilitates receptor sorting into the recycling pathway

The interaction of p-opioid receptor （MOPr） with the neuronal membrane glycoprotein M6a is known to facilitate MOPr endocytosis in human embryonic kidney 293 （HEK293） cells. To further study the role of M6a in the post-endocytotic sorting of MOPr, we investigated the agonist-induced co-internalization of MOPr and M6a and protein targeting after internalization in HEK293 cells that co-expressed HA-tagged MOPr and Myc-tagged M6a. ,We found that M6a, MOPr, and Rab 11, a marker for recycling endosomes, co-localized in endocytotic vesicles, indicating that MOPr and M6a are primarily targeted to recycling endosomes after endocytosis. Furthermore, co-expression of M6a augmented the post-endocytotic sorting of 6-opioid receptors into the recycling pathway, indicating that M6a might have a more general role in opioid receptor post-endocytotic sorting. The enhanced post-endocytotic sorting of MOPr into the recycling pathway was accompanied by a decrease in agonist-induced receptor down-regulation of M6a in co-expressing cells. We tested the physiological relevance of these findings in primary cultures of cortical neurons and found that co-expression of M6a markedly increased the translocation of MOPrsfrom the plasma membrane to intracellular vesicles at steady state and significantly enhanced both constitutive and agonist-induced receptor endocytosis. In conclusion, our results strongly indicate that M6a modulates MOPr endocytosis and post-endocytotic sorting and has an important role in receptor regulation.

Radioiodine therapy for castration-resistant prostate cancer following prostate-specific membrane antigen promoter-mediated transfer of the human sodium iodide symporter

Radioiodine therapy, the most effective form of systemic radiotherapy available, is currently useful only for thyroid cancer because of the thyroid-specific expression of the human sodium iodide symporter （hNIS）. Here, we explore the efficacy of a novel form of gene therapy using prostate-specific membrane antigen （PSMA） promoter-mediated hNIS gene transfer followed by radioiodine administration for the treatment of castration-resistant prostate cancer （CRPC）. The androgen-dependent C33 LNCaP cell line and the androgen-independent C81 LNCaP cell line were transfected by adenovirus. PSMA promoter-hNIS （Ad.PSMApro-hNIS） or adenovirus.cytomegalovirus-hNIS containing the cytomegalovirus promoter （Ad.CMM-hNIS） or a control virus. The iodide uptake was measured in vitro. The in vivo iodide uptake by C81 cell xenografts in nude mice injected with an adenovirus carrying the hNIS gene linked to PSMA and the corresponding tumor volume fluctuation were assessed. Iodide accumulation was shown in different LNCaP cell lines after Ad.PSMApro-hNIS and Ad.CMV-hNIS infection, but not in different LNCaP cell lines after adenovirus.cytomegalovirus （Ad.CMV） infection. At each time point, higher iodide uptake was shown in the C81 cells infected with Ad.PSMApro-hNIS than in the C33 cells （P 〈 0.05）. An in vivo animal model showed a significant difference in 1311 radioiodine uptake in the tumors infected with Ad.PSMApro-hNIS, Ad.CMV-hNIS and control virus （P 〈 0.05） and a maximum reduction of tumor volume in mice infected with Ad.PSMApro-hNIS. These results show prostate-specific expression of the hNIS gene delivered by the PSMA promoter and effective radioiodine therapy of CRPC by the PSMA promoter-driven hNIS transfection.

In situ tumor vaccination with adenovirus vectors encoding measles virus fusogenic membrane proteins and cytokines

AIM： To evaluate whether intratumoral expression of measles virus fusogenic membrane glycoproteins H and F （MV-FMG）, encoded by an adenovirus vector Ad.MV-HI F, alone or in combination with local coexpression of cytokines （IL-2, IL-12, IL-18, IL-21 or GM-CSF）, can serve as a platform for inducing tumor-specific immune responses in colon cancer.METHODS： We used confocal laser scanning microscopy and flow cytometry to analyze cell-cell fusion after expression of MV-FMG by dye colocalization. In a syngeneic bilateral subcutaneous MC38 and Colon26 colon cancer model in C57BL/6 and BALB/c mice, we assessed the effect on both the directly vector-treated tumor as well as the contralateral, not directly vector- treated tumor. We assessed the induction of a tumorspecific cytotoxic T lymphocyte （CTL） response with a lactate dehydrogenase （LDH） release assay.RESULTS： We demonstrated in vitro that transduction of MC38 and Colon26 cells with Ad.MV-H/F resulted in dye colocalization, indicative of cell-cell fusion, in addition, in the syngeneic bilateral tumor model we demonstrated a significant regression of the directly vector-inoculated tumor upon intratumoral expression of MV-FMG alone or in combination with the tested cytokines. We observed the highest anti-neoplastic efficacy with MV-FMG and lL-21 coexpression. The degree of tumor regression of the not directly vector-treated tumor correlated with the anti-neoplastic response of the directly vector-treated tumor. This regression was mediated by a tumor-specific CTL response.CONCLUSION： Our data indicate that intratumoral expression of measles virus fusogenic membrane glycoproteins is a promising tool both for direct tumor treatment as well as for tumor vaccination approaches that can be further enhanced by cytokine coexpression.

原发与继发免疫性血小板减少症患者血小板膜糖蛋白特异性抗体及其与出血评分关系的研究

目的探讨原发与继发免疫性血小板减少症(ITP)患者抗GPⅡb/Ⅲa及GPΙb/Ⅸ抗体的表达、抗体阳性表达与出血评分的关系,以及不同抗体类型出血评分的差异,为原发与继发ITP患者的诊治和出血严重程度提供临床依据。方法选取2013年10月—2020年8月在新疆医科大学第一附属医院诊断为原发ITP患者(42例)及继发ITP患者(42例)为研究对象,所有患者入院时采用ITP出血评分量表行出血评分。应用改良MAIPA法检测患者抗GPⅡb/Ⅲa和抗GPΙb/Ⅸ抗体。结果原发与继发组患者抗GPⅡb/Ⅲa抗体、抗GPΙb/Ⅸ抗体阳性率比较,差异无统计学意义(P>0.05),原发组患者抗体表达以抗GPⅡb/Ⅲa抗体为主,继发组抗体表达以抗GPΙb/Ⅸ抗体为主,两者抗体阳性构成比较,差异有统计学意义(P<0.05)。继发组患者整体出血程度较原发组重(P<0.05)。抗体阳性表达与出血程度的关系:(1)抗GPⅡb/Ⅲa抗体阳性患者出血程度较抗体阴性患者重(P<0.05)。(2)抗GPΙb/Ⅸ抗体阳性患者出血程度较抗体阴性患者重(P<0.05),双抗体阳性患者出血程度较抗体阴性患者重(P<0.05)。结论抗GPⅡb/Ⅲa抗体、抗GPΙb/Ⅸ抗体阳性表达对原发与继发ITP无鉴别意义,但原发ITP患者抗体表达以抗GPⅡb/Ⅲa抗体为主,继发ITP患者抗体表达以抗GPΙb/Ⅸ抗体为主。继发ITP患者临床出血程度较原发ITP患者重。抗GPⅡb/Ⅲa抗体、抗GPΙb/Ⅸ抗体及抗体双阳性患者出血程度较抗体阴性患者重。

龙柴降血方对原发性血小板增多症患者凝血功能及血小板活化蛋白的影响

目的观察龙柴降血方对原发性血小板增多症(ET)患者凝血功能及血小板活化蛋白的影响。方法选取2020年1月—2021年10月中国中医科学院西苑医院收治的ET患者22例,给予龙柴降血方治疗4个月,观察治疗前后血栓弹力图指标[凝血时间(R)、凝血速率(K)、纤维蛋白功能(Angle)、血栓最大振幅(MA)、血栓最大振幅时间(TMA)、即时振幅(A)、30 min振幅(A30)、从血样开始检测到曲线分叉所需时间(SP)、血凝块溶解时间(CLT)]、高凝状态发生率和积分变化。选取其中15例ET患者,采用平行反应监测(PRM)技术检测患者治疗前后血小板活化蛋白[血小板膜糖蛋白Ibα(GPIbα)、糖蛋白VI(GPVI)、选择素-P(SELP)]表达水平,分析比较血小板聚集改善患者和血小板聚集增强患者上述各蛋白表达水平的差异;取5例健康者和10例ET患者治疗前后血清,采用Western blot法检测血清GPIbα表达情况;分析15例ET患者MA与血清GPIbα表达水平的相关性。结果与治疗前比较,龙柴降血方治疗后MA、CLT、高凝状态发生率、高凝状态积分和血清GPIbα、GPVI表达水平均明显降低(P均<0.05),血清SELP表达水平无明显变化(P>0.05)。治疗后血小板聚集改善患者血清GPIbα表达水平明显低于血小板聚集增强患者(P<0.05),GPVI表达水平无明显变化(P>0.05)。与健康人相比,10例ET患者治疗前血清GPIbα相对表达量明显增高(P<0.05);10例ET患者治疗后血清GPIbα相对表达量较治疗前明显降低(P<0.05)。龙柴降血方治疗后MA下降与GPIbα表达下调呈正相关(r=0.6986,P<0.05)。结论龙柴降血方能够抑制ET患者血小板聚集,改善高凝状态,其机制可能与抑制GPIbα蛋白介导的血小板活化信号通路有关。

后节小眼畸形-视网膜劈裂-视网膜玻璃膜疣综合征一家系临床表型和基因型分析

目的分析后节小眼畸形-视网膜劈裂-视网膜玻璃膜疣综合征一家系的临床表型和基因型。方法采用家系调查研究方法,收集2021年7月于深圳市眼科医院就诊的来自中国惠州地区汉族后节小眼畸形综合征一家系共2代4人的临床资料,对各家系成员进行详细的眼科检查,包括最佳矫正视力、眼压、裂隙灯显微镜、彩色眼底照相、光学相干断层扫描(OCT)、眼前节OCT、荧光素眼底血管造影(FFA)和视野检查。采集该家系成员外周静脉血,进行全外显子组测序和数据分析,应用ACMG指南对新发现的变异位点进行致病性分析。结果先证者女,14岁,自幼高度远视,视力右眼+9.75 DS-0.75 DC×150°=0.9,左眼+11.75 DS-1.25 DC×30°=0.7。角膜横径分别为12.1和12.2 mm,前房深度分别为2.56和2.92 mm,晶状体厚度分别为3.92和3.94 mm,眼轴长度分别为17.47和17.01 mm。彩色眼底照相显示中周部视网膜弥漫性分布边界不清的黄白色玻璃膜疣样病灶;OCT显示视网膜内核层劈裂,视网膜色素上皮下多个均质土堆状隆起和高反射致密点;FFA显示双眼中周部视网膜弥漫性斑点状透见荧光;视野检查显示双眼视觉灵敏度总体降低。先证者胞弟8岁,体征与其相似。父母近亲结婚,表型正常。全外显子测序结果显示,先证者及其胞弟膜型卷曲相关蛋白(MFRP)基因第5、10外显子上分别有c.1150_1151insC(p.His384Profs*8)和c.498_499insC(p.Asn167Glnfs*34)2个复合杂合变异,先证者父亲携带c.498_499insC,母亲携带c.1150_1151insC。两者均为移码变异,均可导致基因功能的改变。该家系确定的2个变异位点在ESP数据库、千人数据库(Phase3)、ExAC数据库中未见报道,为新发变异,变异与疾病共分离。根据ACMG指南,以上2个位点的变异均被判断为致病变异。根据临床表型和基因型结果,该家系符合常染色体隐性遗传方式,诊断为后节小眼畸形-视网膜劈裂-视网膜玻璃膜疣综合征。结论MFRP基因c.1150_1151insC和c.498_499insC为该后节小眼畸形-视网膜劈裂-视网膜玻璃膜疣综合征家系的致病变异位点,该复合杂合变异为首次报道。

体外膜氧合联合介入取栓救治蛋白C基因突变所致高危肺栓塞一例

遗传性蛋白C缺陷症是由蛋白C基因突变引起的一种染色体遗传病,可导致静脉血栓形成,多与外显子4~9和内含子8的突变有关。蛋白C基因突变引起的致死性肺栓塞罕见,治疗面临巨大挑战。本文报道1例由蛋白C基因8号外显子移码突变引起的致死性肺栓塞,采用体外膜氧合进行呼吸、循环支持,并成功实行介入取栓的救治经验,为该疾病的诊断及救治提供参考。

慢性非缺血性心力衰竭患者血小板PAC-1和CD62p的表达及其与心功能的关系

目的探讨慢性非缺血性心力衰竭患者血小板膜糖蛋白纤维蛋白原受体(PAC-1)和P选择素(CD62p)的表达及其与心功能的关系。方法选取2019年1月至2022年1月天津市南开医院收治的185例慢性非缺血性心力衰竭患者作为观察组,根据纽约心脏病学会(NYHA)心功能分级将观察组患者分为心功能Ⅱ级组58例、心功能Ⅲ级组88例和心功能Ⅳ级组39例,根据左室射血分数(LVEF)将观察组患者分为射血分数保留心衰组(HFpEF)123例、射血分数临界值心衰组(HFmrEF)25例和射血分数下降心衰组(HFrEF)37例,再以B型钠尿肽(BNP)中位数为界将观察组患者分为高BNP组109例和低BNP组76例;另选取同期心功能正常的就诊者55例作为对照组。比较各组患者的基本临床信息、生化、BNP指标和PAC-1、CD62p的表达水平,采用Pearson相关分析法分析PAC-1、CD62p水平与BNP、LVEF的关系。结果观察组患者的PAC-1、CD62p水平分别为(23.67±16.54)%、(10.43±9.19)%,明显高于对照组的(9.33±9.63)%、(4.24±5.49)%,差异均有统计学意义(P<0.05);不同心功能分级患者对比发现,PAC-1、BNP水平为Ⅳ级>Ⅲ级>Ⅱ级>对照组,CD62p水平为Ⅳ级>Ⅲ级、Ⅱ级>对照组,差异均有统计学意义(P<0.05);不同LVEF分级患者对比发现,PAC-1水平为HFrEF组>HFmrEF组、HFpEF组>对照组,BNP水平为HFrEF组>HFmrEF组>HFpEF组>对照组,HFpEF组、HFmrEF组、HFrEF组CD62p水平分别为(10.72±9.17)%、(9.87±11.19)%、(9.84±7.88)%,明显高于对照组的(4.24±5.49)%,差异均有统计学意义(P<0.05);高BNP组患者的PAC-1、CD62p分别为(26.97±16.94)%、(11.05±9.62)%,明显高于低BNP组的(13.80±12.82)%、(6.97±7.54)%,差异均有统计学意义(P<0.05);经Pearson相关分析结果显示,PAC-1、CD62p与BNP呈正相关(r=0.363、0.182,P<0.05),PAC-1与LVEF呈负相关(r=-0.297,P<0.05)。结论慢性非缺血性心力衰竭患者的PAC-1、CD62p水平均明显增高,且与心功能有关,检测其水平有助于为患者的病情评估及临床治疗提供参考依据。

重组人糖蛋白激素β5/α2融合蛋白在CHO-S细胞中的表达纯化及功能活性分析

目的在悬浮中国仓鼠卵巢细胞(Chinese hamster ovary cells,CHO-S)中分泌表达、纯化重组hCGH-CTP融合蛋白,验证其对3T3-L1成熟脂肪细胞脂质积累的影响。方法构建CTP连接肽融合人糖蛋白激素β5/α2重组蛋白表达载体pcDNA3.1-rhCGH-CTP,将其瞬时转染CHO-S悬浮细胞中,大量表达纯化并验证rhCGH-CTP蛋白生物学活性;通过干预3T3-L1成熟脂肪细胞24 h,观察细胞内甘油三酯(TG)水平的变化。结果Western blot结果显示,rhCGH-CTP蛋白在CHO-S细胞中成功表达,表达量可达715.4 mg·L^(-1);用AKTA pure蛋白纯化系统纯化蛋白,SDS-PAGE方法鉴定纯化出的蛋白纯度较高可达90%。此外,在高表达TSHR基因的成熟脂肪细胞3T3-L1中,利用ELISA试剂盒测定不同浓度rhCGH-CTP蛋白干预后胞内cAMP含量明显升高,说明rhCGH-CTP蛋白具有生物活性;油红O染色结果发现,与对照组相比,不同浓度rhCGH-CTP蛋白干预组的成熟脂肪细胞中TG含量明显降低(P<0.05)。结论成功表达并纯化了rhCGH-CTP融合蛋白,其具有良好的生物学活性并能有效降低TG,该研究为后续深入揭示CGH蛋白的生理作用及在临床实践中的潜在应用提供了重要基础。

人类白细胞抗原-DQA1、细胞色素P4503A5*3基因多态性与特发性膜性肾病遗传易感性关联性研究

目的:探讨分析人类白细胞抗原(HLA)-DQA1、细胞色素P450(CYP)3A5*3基因多态性与特发性膜性肾病(IMN)遗传易感性的关联性。方法:将IMN患者62例作为研究组,另将健康者62例作为对照组。采用聚合酶链反应(PCR)技术和基因测序技术检测入组患者的HLA-DQA1(rs2187668)、CYP3A5*3(rs776746)基因型,判断各基因型与IMN患者的关系。结果:研究组中HLA-DQA1 rs2187668基因型AA占比高于对照组(P<0.05),基因型GG、AG占比低于对照组(均P<0.05);研究组等位基因A占比高于对照组(P<0.05)。研究组与对照组CYP3A5*3 rs776746不同基因型占比及等位基因占比比较无统计学差异(均P>0.05)。HLA-DQA1 rs2187668AA基因型IMN患者24 h尿蛋白量和血肌酐水平显著高于AG和GG型,肾小球滤过率显著低于AG和GG型(均P<0.05)。结论:HLA-DQA1 rs2187668位点的基因多态性与IMN的发生有关,AA基因型和等位基因A可增加IMN易感性。

Deletion of the first glycosylation site promotes Lassa virus glycoprotein-mediated membrane fusion

The Lassa virus(LASV)is endemic in West Africa and causes severe hemorrhagic Lassa fever in humans.The glycoprotein complex(GPC)of LASV is highly glycosylation-modified,with 11 N-glycosylation sites.All 11 N-linked glycan chains play critical roles in GPC cleavage,folding,receptor binding,membrane fusion,and immune evasion.In this study,we focused on the first glycosylation site because its deletion mutant(N79Q)results in an unexpected enhanced membrane fusion,whereas it exerts little effect on GPC expression,cleavage,and receptor binding.Meanwhile,the pseudotype virus bearing GPC_(N79Q)was more sensitive to the neutralizing antibody 37.7H and was attenuated in virulence.Exploring the biological functions of the key glycosylation site on LASV GPC will help elucidate the mechanism of LASV infection and provide strategies for the development of attenuated vaccines against LASV infection.

GPIa C807T和G873A基因多态性对血小板聚集抑制率的影响

目的:探讨GPIa C807T和G873A基因多态性对血小板聚集抑制率的影响,为临床个体化抗血小板聚集治疗提供实验数据支持。方法:收集正在使用阿司匹林抗血小板聚集治疗的住院患者80例,采用血栓弹力图仪检测血小板聚集抑制率,根据聚集抑制率检测结果分为阿司匹林抵抗(aspirin resistance,AR)组(共10例)和阿司匹林敏感(aspirin sensitivity,AS)组(共70例),分析GPIa C807T、G873A基因型及其等位基因在两组分布状况以及两者患者临床资料与实验检测指标关系。结果:AR组807T、873G等位基因高于AS组(χ^(2)=4.039、4.354,P=0.044、0.037);AR组高血压病史患病率、卒中史高于AS组(χ^(2)=4.612、4.737,P=0.032、0.030);AR组血浆D-二聚体(D-D)、血清甘油三酯(TG)高于AS组(t=2.499、2.276,P=0.016、0.033),而AR组血小板聚集抑制率、血清葡萄糖低于AS组(t=15.352、3.076,P<0.001、0.005)。结论:GPIa C807T和G873A基因多态性与AR存在相关性,建议联合检测两基因位点的多态性并结合血小板聚集抑制率,共同指导临床个体化抗血小板聚集治疗。

中空纤维膜基蒸发水冷器的多目标优化

提出一种中空纤维膜基蒸发水冷器,以解决传统水侧蒸发冷却系统的空气-水交叉污染和液滴飘逸等问题,并对逆流式中空纤维膜基蒸发水冷器的关键设计参数进行优化研究。在开式冷却塔的标准工况下,通过数值模拟和响应面法获得2个回归模型,建立水流速、气水比、纤维长度、纤维内径、膜层厚度和填充率6个设计参数与出口水温和耗电比2个性能评价指标之间的关联性。通过方差分析验证模型的可靠性,即R2超过0.98,信噪比高于45.92。采用遗传算法同时最小化2个评价指标,得到一组最佳参数组合,构成帕累托前沿。此外,通过理想点法确定相对最优解,结果表明:优化方案能够使中空纤维膜基蒸发水冷器的出口水温和耗电比分别达到29.44℃和0.0128kW·h/m^(3),相比原始设计分别降低10.23%和94.52%。最后通过对帕累托前沿的验证所提出的优化方案在工程实践中的可靠性。