弓形虫致密颗粒蛋白GRA2真核表达质粒的构建与体外表达

目的构建弓形虫致密颗粒蛋白GRA2的真核表达重组质粒。方法设计合成GRA2引物，运用PCR方法扩增其基因片段，经克隆至pMDl8-T载体后，亚克隆至真核表达质粒pcDNA3．1（-）而构建重组表达质粒pcDNA3．1-GRA2。脂质体法将构建的重组质粒转染HFF细胞，RT—PCR法检测转染细胞中GRA2的表达情况。结果PCR扩增GRA2基因序列正确，构建的重组表达质粒pcDNA3．1-GRA2经PCR、EcoRⅠ／HindⅢ双酶切和测序鉴定正确；转染GRA2基因的细胞，RT—PCR可见目的条带。结论成功获得真核表达重组质粒pcDNA3．1-GRA2，为进一步研究弓形虫疫苗的免疫保护性奠定基础。     

桑仙降压颗粒对自发性高血压大鼠心肌VEGF及nm23的干预

[目的]观察桑仙降压颗粒对自发性高血压大鼠(SHR)心肌血管内皮生长因子(VEGF)及nm23的影响,探讨其对高血压病心肌血管生长促进因子、抑制因子的干预作用。[方法]SHR随机分为3组,空白对照组、吲哒帕胺西药对照组、桑仙降压颗粒组,以Wistar大鼠为正常对照,4周及8周后观察SHR心肌VEGF及nm23的表达。[结果]实验前正常对照组与空白对照组相比VEGF及nm23蛋白光密度值差异均无统计学意义。实验4、8周后相较空白对照组,桑仙降压颗粒组VEGF表达明显增多而nm23蛋白密度值显著减少。[结论]桑仙降压颗粒能增强VEGF的表达及降低nm23的蛋白密度值。     

Toujie Quwen granule used with conventional western therapy for coronavirus disease 2019: A protocol for systematic review and meta-analysis

Background:Coronavirus disease 2019 (COVID-19) is an epidemic infectious disease resulted from 2019 novel coronavirus (2019-nCoV). Up till now, COVID-19 has swept globally. Currently, due to many high-profiled benefits, clinical studies on Toujie Quwen granule (TJQW) have been increasing. The aim of the study is to assess the efficacy and safety of TJQW used with conventional western therapy for COVID-19.Methods:Relevant randomized controlled trials (RCTs) were searched in Chinese and English databases, and the search time is January 2020 to May 2021. English databases include PubMed, Embase, Web of Science, and the Cochrane Library. Chinese databases include CNKI, WF, VIP, and CBM. The international clinical trial registration platform and the Chinese clinical trial registration platform of controlled trials will be searched by us from January 2020 to May 2021. According to the inclusion and exclusion criteria, screening literature, extraction data will be conducted by 2 researchers independently. Statistical analysis will be conducted using the RevMan 5.3.5 software. After screening the literature based on the inclusion and exclusion criteria, The Recommendation, Assessment, Development, and Evaluation (GRADE) system will be used to evaluate the quality of each result.Results:This study will provide the evidence for TJQW to be used with conventional western therapy for COVID-19.Conclusion:The efficacy and safety of TJQW used with conventional western therapy for COVID-19 will be assessed.INPLASY registration number:INPLASY202150038     

刚地弓形虫RH株GRA6分子基因克隆与序列分析

目的　克隆刚地弓形虫 RH株致密颗粒抗原 (Dense granule antigen,GRA6 )分子基因 ,为研究使用重组的GRA6分子作为抗原进行弓形虫病诊断奠定基础。　方法　根据己知的 GRA6序列合成一对引物 ,抽提弓形虫速殖子m RNA,以速殖子 m RNA为模板 ,通过反转录 PCR(RT- PCR)扩增出 GRA6的 c DNA条带。目的基因 DNA条带被克隆到质粒 p UC19中形成重组质粒。对重组质粒 DNA进行纯化 ,并对目的基因片段进行核苷酸序列分析。　结果　通过RT- PCR从 m RNA中扩增出一条分子量约 70 0 bp的 DNA条带。核苷酸序列分析表明 ,GRA6分子基因的开放的阅读框全长为 6 93bp,编码 2 30个氨基酸分子 ,与已报道的 RH株 GRA6分子具有 10 0 %的同源性。　结论　本研究获得了刚地弓形虫 RH株的 GRA6分子基因 ,为今后应用此分子作为抗原诊断弓形虫病奠定了基础     

血尿冲剂治疗慢性肾小球源性血尿

目的 观察血尿冲剂治疗肾小球源性血尿(肾性血尿)的疗效.方法 对40例确诊为肾性血尿的患者采用血尿冲剂(穿山甲粉、琥珀粉、花蕊石粉等)治疗,8周为1个疗程,比较治疗前后尿常规指标变化和尿常规各项指标的动态变化.结果 治疗1个疗程后,临床控制27例(67.5％),显效6例(15％),有效5例(12.5％),无效2例(5％),总有效率95％,对蛋白尿有一定的改善.结论 血尿冲剂治疗肾性血尿,能活血通络,化瘀止血,使瘀血祛,经络通,出血止,血尿愈,对尿蛋白亦有一定的改善.     

益母草颗粒剂的制备工艺研究

目的:探讨益母草颗粒剂的制备工艺.方法:用正交法确定益母草颗粒剂的最佳制备工艺条件,在此基础上,加入新型的甜味剂,制成颗粒剂.结果:在益母草水煎液、浸膏剂型的基础上加入适量的糊精和罗汉果苷,以80%的乙醇溶液为湿润剂制成颗粒,干燥,便可得到益母草颗粒剂. 结论:用这种方法制成的益母草颗粒剂质量稳定,更加安全,可以用此制备工艺进行大批量生产.     

妇科万应膏治疗宫寒血瘀型痛经的临床效果观察

目的：观察妇科万应膏治疗宫寒血瘀型痛经患者的疗效和安全性。方法根据纳入/排除标准,共筛选痛经患者223例,采用区组随机的方法,按照3：1的比例随机分为试验组167例与对照组56例。试验组使用妇科万应膏外用穴位贴敷,对照组使用痛经宝颗粒口服,共用药3个月经周期。治疗后比较2组的治疗效果与不良反应。结果治疗3个月经周期后,试验组总有效率为91.02%明显优于对照组的78.57,差异有统计学意义（P ＜0.05）。试验组发生不良反应5例,不良反应发生率2.89%;对照组无不良反应发生,组间比较差异无统计学意义（ P ＞0.05）。结论妇科万应膏治疗宫寒血瘀型月经不调和痛经安全有效,值得临床推广使用。     

Expression of reg protein in rat regenerating islets and its co-localization with insulin in the Beta cell secretory granules

Summary Regenerating islets can be induced by the administration of poly(ADP-ribose) synthetase inhibitors to 90% depancreatized rats. In screening a regenerating isletderived cDNA library, we previously isolated a novel gene, reg (regenerating gene), which encodes a 165-amino acid protein with a 21-amino acid signal sequence. In the present study, we have examined the expression and localization of reg protein in the regenerating islets by immunocytochemical techniques using a monoclonal antibody against a recombinant rat reg protein of 144 amino acids without the signal sequence. Light microscopy examination showed strong immunoreactivity for reg protein in the regenerating islets of the rats at two weeks and two months after the 90% pancreatectomy, whereas reg protein was almost undetectable in normal rat islets or in the islets of the rats one year after the pancreatectomy. Almost all the reg protein-positive cells were stained for insulin. By applying the immunogold technique at the ultrastructural level, it was demonstrated that both reg protein and insulin occur in the central granular core of the regenerating Beta cell secretory granules. These results suggest that reg protein is synthesized in and secreted from the regenerating Beta cells and that its expression is closely associated with Beta-cell regeneration.     

灵葜四君颗粒联合化疗治疗胃癌的临床研究

目的:观察灵葜四君颗粒联合化疗治疗胃癌的临床疗效.方法:80例胃癌术后患者随机分为治疗组(灵葜四君颗粒加化疗)和对照组(单纯化疗)各40例.观察治疗前后卡氏评分、外周血常规和血液流变学的变化,并观察自然杀伤(NK)细胞、T淋巴细胞亚群、可溶性白细胞介素-2受体(SIL-2R)等指标的变化.结果:治疗组化疗后生存质量改善,外周血白细胞和红细胞数稳定;对照组外周血白细胞、红细胞数、血红蛋白浓度均较化疗前明显下降.治疗组化疗后NK细胞活性、CD3、CD4阳性T细胞数和CD4/CD8比值升高,血清SIL-2R的浓度和全血粘度(低切、中切、高切)均明显下降;而对照组化疗后NK细胞活性、CD4阳性T细胞数和CD4/CD8比值下降,血清SIL-2R的浓度升高而全血粘度无明显改变.结论:灵葜四君颗粒对化疗有增效减毒作用,对胃癌术后患者机体免疫功能有保护和促进作用,并能防治化疗引起的骨髓抑制,降低血液粘度.     

Anti-eosinophil peroxidase antibodies detected in patients with primary biliary cirrhosis.

OBJECTIVE:: The possible involvement of eosinophils in primary biliary cirrhosis (PBC) has been suggested for many years. Activated eosinophils release basic granule proteins and are thought to be involved in the tissue damage of PBC. Eosinophil peroxidase (EPO) is one of the eosinophilic granule proteins and is toxic to epithelial cells. In this study, we examined whether autoantibodies to EPO are present in sera of patients with PBC and we evaluated the relationship between the presence of anti-EPO antibodies and the clinical features of PBC. METHODS:: Sera obtained from 61 patients with PBC, 31 patients with autoimmune hepatitis (AIH), 87 patients with chronic viral hepatitis (CVH), 27 patients with bronchial asthma, and 20 healthy controls were examined. Enzyme-linked immunosorbent assay (ELISA) and Western blot methods were used for detection of anti-EPO antibodies. RESULTS:: The mean OD value of anti-EPO antibodies, as determined by ELISA, was significantly (p<0.01) higher in patients with PBC (0.197+/-0.121) than in patients with AIH (0.124+/-0.077), CVH (0.090+/-0.038) or bronchial asthma (0.073+/-0.025) and in healthy controls (0.072+/-0.029). The results of ELISA showed that 32 (52.5%) of the 61 patients with PBC, 9 (29.0%) of the 31 patients with AIH, 7 (8.0%) of the 87 patients with CVH, and 1 (3.7%) of the 27 patients with bronchial asthma were positive for anti-EPO antibodies. In addition, PBC patients who were positive for anti-EPO antibodies had a significantly smaller number of peripheral eosinophils than did patients who were negative for anti-EPO antibodies (99.2+/-54.4cells/mul versus 176.9+/-117.5cells/mul, p<0.01). However, there was no correlation between the titers of anti-EPO antibodies and those of AMA or AMA-M2, ANA or serum levels of IgM. CONCLUSIONS:: This is the first report of the detection of anti-EPO antibodies in patients with PBC. Further study is needed to clarify the role of anti-EPO antibodies in the pathogenesis of PBC.