Developing and piloting an expert system for better routine voluntary HIV counseling and testing in China: preliminary results and lessons

This study aims at developing and testing a pragmatic expert system for HIV voluntary counseling (VCT-ES) that leverages best practices. The VCT-ES was developed via evolutionary prototyping and piloted in 10 voluntary HIV counseling and testing (VCT) clinics from Anhui and Beijing representative of prefecture and county level VCT clinics in China. All counselors with the clinics and the clients to the clinics within selected two weeks at baseline and the end of study were invited to participate. Assessment measures included essential counseling procedures (ECPs); clients' satisfaction, knowledge, and behavior efficacy. VCT-ES was developed which tries to model and facilitate standard VCT operation procedures and best practices. One hundred and eighty-two (96 at baseline vs. 86 after intervention) cases recordings, 172 (96 vs. 76) client questionnaires, 10 counselor, and 2 expert rating instruments were collected; and 17 clients and 8 counselors participated in qualitative interviews. VCT-ES increased delivery of ECPs from 18.94% to 66.39% on average; increased clients' knowledge from 40.51% to 86.34% and self-efficacy by 22.42%. The VCT-ES applications listed were rated 9.1 on average (maximum = 10). The VCT-ES could be an easy and effective solution to better routine VCT and merits further research.     

Plasma level of lipopolysaccharide-binding protein is indicative of acute graft-versus-host disease following allogeneic hematopoietic stem cell transplantation

Acute graft-versus-host disease (aGVHD) is the major cause of non-relapse mortality following allogeneic hematopoietic stem cell transplantation. To date, there are no consensus specific plasma biomarkers for aGVHD. We recently identified several candidates differentially expressed in aGVHD patients. Here, we have validated one such candidate: lipopolysaccharide-binding protein (LBP). We detected plasma LBP level by ELISA in 73 patients and performed correlation analysis with the progression and severity of aGVHD. We found that plasma LBP level increased during the period of aGVHD and decreased markedly as aGVHD was resolved. LBP level in patients with moderate aGVHD (25-50 % skin rash area of grade 1 and grade 2) was higher than in patients with no, little (skin rash area <25 % of grade 1), or severe aGVHD (grade 3-4). Higher LBP level indicated higher probability of aGVHD. Multivariate analysis showed that LBP level above 15000 ng/ml was significantly associated with an increased risk of aGVHD (HR 2.43; 95 % CI 1.29-4.58; P = 0.006). If LBP level exceeded 15000 ng/ml at d7 and d14 after HSCT, the subsequent probability of aGVHD increased markedly, especially at the time point of d14. There was no correlation between LBP level and the site of aGVHD. In conclusion, our study demonstrated that an elevated LBP level of >15000 ng/ml may serve as a biomarker for the prediction and monitoring of aGVHD.     

刚地弓形虫磷酸甘油酸变位酶2基因片段克隆、表达及抗原性分析

目的克隆、表达刚地弓形虫(Toxoplasma gondii)磷酸甘油酸变位酶2(TgPGAM2)基因片段,并分析其抗原性。方法提取弓形虫RH株速殖子总RNA,逆转录合成cDNA。PCR扩增TgPGAM2基因。扩增产物经双酶切后连接入pET30a(+)载体,重组质粒转化大肠埃希菌(E.coli)DH5α,阳性菌落经PCR和双酶切鉴定,并测序。将测序正确的重组质粒pET30a(+)-TgPGAM2转化至E.coli BL21并加入异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)结合考马斯亮蓝染色检测表达产物。以兔抗弓形虫血清为一抗,蛋白质印迹(Western blotting)分析重组蛋白的抗原性。结果PCR扩增产物约为750 bp。菌落PCR、双酶切和测序结果显示,重组质粒pET30a(+)-TgPGAM2构建成功。SDS-PAGE结果显示,经IPTG诱导获得相对分子质量(Mr)约30 000的可溶性重组蛋白。Western blotting分析证实其能被兔抗弓形虫血清识别。结论刚地弓形虫RH株TgPGAM2基因片段可在原核表达系统中表达,且该可溶性重组蛋白具有抗原性。     

动态心电图对体表心电图P-R间期延长的诊断价值

目的 探讨动态心电图对体表心电图P．R间期延长的诊断价值,分析P—R间期延长的临床意义和预后。方法选择2008年1月至2011年8月在昆明医学院第二附属医院住院和门诊心电图有P-R间期延长的患者90例为研究对象．监测其动态心电图。分析P．R间期的变化及阵发性心房颤动、房室结双径路传导、第二度I型房室阻滞、间歇性左右束支阻滞4类心律失常。并且根据P—R间期延长程度分为3组,对比其与上述4类心律失常的关系。结果动态心动图结果中伴发第二度I型房室阻滞2l例（23．33％）、房室结双径路传导17例（18．89％）、间歇性左右束支阻滞9例（10％）、阵发性心房颤动7例（7．78％）。在所有的患者中,P．R间期延长伴发上述4类心律失常发生率为60％。心电图结果中,P-R间期延长50例,显著延长29例,过度延长11例。动态心动图结果中。P。R间期延长32例,显著延长38例,过度延长20例。结论P—R间期延长不应均视为良性和无须干预的心律失常,相反,需要进行进一步动态心动图监测及电生理检查和追踪观察,争取早期发现病变、消除病变,为临床早期诊断、早期治疗提供辅助手段。     

二维斑点追踪技术评价社区人群左心房应变率与左心室舒张功能的相关性

目的:应用二维斑点追踪技术评价社区人群左心房应变率,及其与左心室舒张功能分级的相关性。方法:研究对象为左心室射血分数正常的社区人群,共708例脱机处理,根据多普勒二尖瓣口血流、二尖瓣环组织、肺静脉血流对左心室舒张功能进行分组:A组(左心室舒张功能正常组)、B组(左心室舒张功能轻度受损组)、C组(左心室舒张功能中/重度受损组)。采用二维斑点追踪技术(STE),测量收缩期左心房应变率峰值SRs,舒张早期左心房应变率峰值SRe和舒张晚期左心房应变率峰值SRa。结果:①B组、C组SRs较A组降低,分别为(1.07±0.35)s-1、(0.90±0.24)s-1和(1.12±0.28)s-1(P<0.01),B组、C组SRe较A组亦明显降低,分别为(-0.79±0.35)s-1、(-0.65±0.29)s-1和(-1.02±0.33)s-1(P<0.01);SRa在B组高于A组[分别为(-1.60±0.58)s-1和(-1.48±0.47)s-1,P<0.05],亦高于C组[分别为(-1.60±0.58)s-1和(-1.24±0.48)s-1,P<0.01],且C组显著小于A组[分别为(-1.24±0.48)s-1和(-1.48±0.47)s-1,P<0.01]。②SRs、SRe与左心室舒张功能分级均呈负相关(r分别为-0.178和-0.366,P<0.01)。结论:二维斑点追踪技术测量左心房应变率,可间接反映左心室舒张功能。     

P2X_1受体在急性冠状动脉综合征中的表达

目的观察P2X1受体在急性冠状动脉综合征(ACS)患者血小板中的表达及激活P2X1受体后血小板钙离子浓度的变化,探讨P2X1受体在ACS发生时对血小板功能的调节作用。方法选取70例ACS患者为研究对象,其中急性心肌梗死(AMI)患者30例,不稳定型心绞痛(UA)患者40例,同期入院的健康受试者20例为对照组。应用反转录-聚合酶链反应(RT-PCR)和实时荧光定量PCR方法,观察P2X1受体和血小板膜糖蛋白Ⅱb/Ⅲa(GPⅡb/Ⅲa)受体在对照组和ACS组血小板中的表达情况。以α,β-me ATP(3μmol/L)激活血小板,Fluo-3 AM Ester为标记物在荧光分光光度计下观察血小板钙离子水平变化。结果对照组及ACS组的血小板中均有P2X1受体、GPⅡb/Ⅲa受体的表达,且在ACS组的表达较对照组高(1.62±0.69比0.88±0.04,1.58±0.11比0.85±0.06,P<0.05);ACS组患者钙离子基线水平高于对照组[(418.96±31.85)nmol/L比(307.20±23.37)nmol/L,P<0.05]。结论 P2X1受体在ACS的发生发展过程中起重要作用。     

脂联素受体在结直肠癌和结直肠腺瘤组织中表达

目的:检测脂联素受体1(adiponectin receptors1,R1)和R2在人结直肠癌组织、腺瘤组织、癌旁组织中的表达情况与临床病理特征的关系及3种组织中R1和R2的关联性.方法:采用免疫组织化学SP法检测51例新鲜结直肠癌组织,42例结直肠腺瘤组织,35例癌旁组织中脂联素受体R1和R2的表达.结果:免疫组织化学结果显示R1在人结直肠癌组织中表达高于腺瘤组织(P=0.047),结直肠癌组织中表达高于癌旁组织(P=0.002),腺瘤组织中的表达高于癌旁组织(P=0.042).R2在人结直肠癌组织中表达高于腺瘤组织(P=0.035),结直肠癌组织中表达高于癌旁组织(P=0.002),腺瘤组织中表达高于癌旁组织(P=0.046).在3种组织中,R1与R2的表达无关联性(P>0.05).在51例结直肠癌组织中脂联素受体表达与分化程度、淋巴结转移、TNM分期相关(均P<0.05),与性别、年龄、肿瘤大小、组织学类型无明显相关(P>0.05).结论:脂联素受体在人结直肠癌组织、腺瘤组织、癌旁组织中普遍表达,且在结直肠癌组织中的表达最高,腺瘤次之.脂联素受体表达与分化程度、淋巴结转移、TNM(tumor regional lymph node and metastasis)分期有关.脂联素受体有可能成为结直肠癌治疗的一个新靶点.     

鸟苷酸环化酶C基因沉默对人胃癌裸鼠皮下种植瘤的影响

目的:探讨鸟苷酸环化酶C(GC-C)基因沉默对人胃癌裸鼠皮下种植瘤的影响及其机制.方法:将SGC-7901细胞(SGC-7901组)、空质粒转染的细胞(PRNA组)、GC-C基因稳定沉默的细胞(GC-C-shRNA组)悬液先在裸鼠皮下接种成瘤,再取瘤组织块分别接种到裸鼠皮下,建立3种胃癌裸鼠皮下种植瘤模型.观察裸鼠一般情况、肿瘤的成瘤率、生长速度,描绘肿瘤的生长曲线,计算肿瘤的抑瘤率;采用实时荧光定量PCR(RFQ-PCR),Western blot和免疫组织化学法检测GC-C和趋化因子受体4(CXCR4)在各组移植瘤组织中的表达.结果:与SGC-7901组和PRNA组比较,GC-C-shRNA组裸鼠移植瘤生长速度明显减慢、体积明显变小(抑制率分别为33.7%、33.2%),肿瘤异型性、坏死程度明显降低,差异具有统计学意义(均P<0.05),且移植瘤组织中GC-C和CXCR4 mRNA和蛋白均明显下调(GC-CmRNA:7.47±1.70 vs 11.18±0.60,11.28±0.85;GC-C蛋白:0.52±0.15 vs 1.04±0.19,1.03±0.24;CXCR4蛋白:0.67±0.13 vs 1.02±0.21,1.03±0.23,均P<0.05).结论:GC-C基因稳定沉默在体内能保持稳定,且能抑制裸鼠移植瘤生长,该过程可能与CXCR4下调有关.     

重度支气管哮喘硫酸镁治疗疗效分析

目的探讨重度支气管哮喘应用硫酸镁治疗疗效与安全性。方法将2009年11月～2011年4月于我院诊治的支气管哮喘重度患者81例分为两组。对照组39例均给予吸氧、平喘等常规治疗;观察组42例在对照组治疗外另给予硫酸镁治疗。结果观察组42例患者显效34例,有效7例,无效1例,总有效率97.62%,显著高于对照组(χ2=12.85,P<0.01)。两组患者均无不良反应。结论对重度支气管哮喘患者在常规治疗外静脉滴注硫酸镁治疗,疗效显著。