补体因子H相关蛋白1促进巨噬细胞分泌肿瘤坏死因子-α调控足细胞增殖和迁移实验研究

目的:探讨补体因子H相关蛋白1(CFHR1)通过巨噬细胞分泌肿瘤坏死因子-α(TNF-α)调控足细胞增殖和迁移的作用。方法:体外培养人单核巨噬细胞和人肾足细胞。巨噬细胞分为对照组和CFHR1干预组,分别进行牛血清白蛋白或CFHR1重组蛋白干预24 h,ELISA法测定上清液TNF-α水平。足细胞分为空白组、TNF-α干预组、对照上清液干预组、CFHR1上清液干预组、CFHR1上清液+TNF-α中和抗体干预组。CCK8法检测各组细胞增殖。Transwell法检测各组细胞迁移。Wb法检测各组细胞中相关蛋白变化。结果:巨噬细胞的CFHR1干预组上清液中TNF-α含量显著增加(P<0.05)。与空白组比较,TNF-α干预组、CFHR1上清液干预组的细胞增殖比率和迁移数量均显著降低(均P<0.05)。与CFHR1上清液干预组比较,CFHR1上清液+TNF-α中和抗体干预组的细胞增殖比率和迁移数量均显著提高(均P<0.05)。与空白组比较,TNF-α干预组、CFHR1上清液干预组的足细胞裂孔膜蛋白(Nephrin)、足突蛋白(Podocin)、纤维状肌动蛋白(F-Actin)、整合素α3β1蛋白(α3β1)表达均显著降低(均P<0.05)。与CFHR1上清液干预组比较,CFHR1上清液+TNF-α中和抗体干预组的Nephrin、Podocin、F-actin、α3β1蛋白表达均显著增多(均P<0.05)。结论:CFHR1促进巨噬细胞分泌的TNF-α可显著抑制足细胞增殖水平和迁移能力,这可能是高浓度CFHR1促进肾病综合征发展的途径。

亚硝酸盐对焦酚红法检测24 h尿液总蛋白的干扰分析

目的探讨亚硝酸盐对于焦酚红法检测24 h尿液总蛋白的干扰效应,并评估维生素C纠正干扰的可行性。方法根据美国临床和实验室标准协会(CLSI)EP7-A3文件,选取24 h尿液总蛋白定量结果为150 mg/L、500 mg/L和1000 mg/L的新鲜尿液标本作为对照样本,并配制含不同浓度亚硝酸钠的干扰物样本。利用配对差异实验确认亚硝酸盐的干扰作用,并通过剂量效应实验明确亚硝酸盐浓度与干扰程度的关系。在终浓度为200μg/mL亚硝酸钠存在的情况下,评估不同浓度维生素C纠正干扰的效果。同时收集61例亚硝酸盐阳性和40例亚硝酸盐阴性的临床标本,比较两组标本维生素C纠正前后的相对差值,评估其临床应用价值。结果配对差异实验中200μg/mL亚硝酸钠对150 mg/L和500 mg/L两个浓度的尿总蛋白测定分别存在-157.8%和-36.2%的相对干扰,显著大于1/2TEa(22%),而对于高浓度1000 mg/L的尿总蛋白虽存在-20.5%的负干扰,但在可接受范围内。剂量效应实验结果显示随着尿液中亚硝酸盐浓度的增加,尿总蛋白检测所受负干扰作用亦逐渐增大。在终浓度为200μg/mL亚硝酸钠存在的情况下,添加0.2 mg/mL的维生素C可分别将150 mg/L和500 mg/L的尿总蛋白纠正至148 mg/L(-1.1%)和402 mg/L(-19.5%),均处于可接受范围内。临床标本中亚硝酸盐阳性组通过维生素C纠正产生的相对差值显著高于亚硝酸盐阴性组(P<0.01)。结论亚硝酸盐对于焦酚红法检测尿液总蛋白存在负干扰,尤其是在尿总蛋白150 mg/L情况下,需引起临床实验室的关注。添加0.2 mg/mL维生素C可有效纠正低浓度尿总蛋白标本中亚硝酸盐的干扰作用,具有一定的临床应用价值。

免疫检查点T细胞激活抑制物免疫球蛋白可变区结构域、人内源性逆转录病毒-H长端重复相关蛋白2在子宫内膜癌中的表达及临床意义

目的分析免疫检查点T细胞激活抑制物免疫球蛋白可变区结构域(VISTA)、人内源性逆转录病毒-H长端重复相关蛋白2(HHLA2)在子宫内膜癌(EC)中的表达及临床意义。方法选取2017年9月至2019年10月唐山市妇幼保健院收治的120例的EC患者作为研究对象,收集其的EC组织和癌旁正常组织。检测EC组织和癌旁正常组织VISTA、HHLA2的表达;采用Spearman分析免疫检查点VISTA、HHLA2的相关性;Kaplan-Meier分析VISTA、HHLA2与预后的关系;Cox回归分析影响EC患者预后的危险因素。结果EC组织中VISTA、HHLA2阳性表达率显著高于正常癌旁组织(P<0.05)。Spearman相关性结果显示EC组织中VISTA与HHLA2表达呈正相关性(r=0.587,P<0.05)。EC组织中VISTA、HHLA2表达与临床病理特征有关(P<0.05)。Kaplan-Meier曲线结果显示VISTA阳性表达患者3年生存率低于阴性表达患者(χ^(2)=11.864,P<0.05),HHLA2阳性表达患者3年生存率低于阴性表达患者(χ^(2)=4.975,P<0.05)。Cox回归分析结果显示VISTA和HHLA2是影响EC患者预后的危险因素(P<0.05)。结论免疫检查点VISTA、HHLA2在EC中高表达,其是影响EC预后的危险因素,二者可能是有价值的预后标志物。

新型Na^(+)/H^(+)逆向转运蛋白UPF0118的Na^(+)结合鉴定

UPF0118蛋白是一种新型Na^(+)(Li^(+))/H^(+)逆向转运蛋白,属于AI-2E超家族,目前已被划分至Na^(+)/H^(+)逆向转运蛋白亚家族,但无直接证据表明该蛋白可与Na+产生相互作用。为鉴定UPF0118蛋白是否具有Na^(+)结合功能,利用PCR技术扩增upf0118基因编码序列,通过Eco RⅠ和PstⅠ酶切位点构建至原核表达载体p ETDuet-1,转化至E. coli BL21*(DE3),对诱导后大量表达的蛋白进行Ni^(2+)亲和层析和凝胶过滤层析,利用等温滴定量热技术对纯化蛋白进行滴定。结果表明,在pH 5.5条件下,Na^(+)滴入导致蛋白溶液产生明显放热现象。在pH 5.5条件下该蛋白具有Na^(+)结合功能,证明该蛋白Na+结合功能为其功能单元与分子机制的研究奠定基础。

绝经后H型高血压合并骨质疏松症患者血清MCP-1、PTX3、GDF15的变化及其临床意义分析

目的分析绝经后H型高血压(HHT)合并骨质疏松症(OP)患者血清单核细胞趋化蛋白(MCP)-1、正五聚蛋白3(PTX3)、生长分化因子15(GDF15)的变化及其临床意义。方法选取2020年6月至2023年5月我院收治的127例绝经后HHT合并OP患者为研究组,另选取同期85例绝经后OP患者为OP组,62例绝经后单纯HHT患者为HHT组、60例体检健康的绝经后女性志愿者为健康组。比较4组血清MCP-1、PTX3、GDF15水平、股骨颈和腰椎(L_(1~4))骨密度、血清骨代谢指标[骨钙素(BGP)、Ⅰ型胶原羧基端肽β特殊序列(β-CTX)、Ⅰ型前胶原氨基端前肽(PINP)]及炎症因子[肿瘤坏死因子α(TNF-α)、白细胞介素-6(IL-6)]。采用Pearson相关性分析血清MCP-1、PTX3、GDF15水平与骨密度、血清骨代谢指标及炎症因子的相关性。采用受试者工作特征(ROC)曲线分析血清MCP-1、PTX3、GDF15对绝经后HHT合并OP的诊断价值。结果与健康组比较,OP组、HHT组及研究组血清MCP-1、PTX3、GDF15、BGP、β-CTX、PINP水平均较高,且OP组高于HHT组,研究组高于OP组和HHT组(P<0.05)。与健康组比较,OP组、HHT组及研究组血清TNF-α、IL-6水平均较高,且HHT组高于OP组,研究组高于OP组和HHT组(P<0.05)。与健康组比较,OP组、HHT组及研究组的股骨颈和腰椎(L_(1~4))骨密度均较低,且OP组低于HHT组,研究组低于OP组和HHT组(P<0.05)。Pearson相关性分析显示,血清MCP-1、PTX3、GDF15水平与股骨颈和腰椎(L_(1~4))骨密度呈负相关,与血清BGP、β-CTX、PINP、TNF-α、IL-6水平呈正相关(P<0.05)。ROC曲线分析显示,血清MCP-1、PTX3、GDF15联合诊断绝经后HHT合并OP的AUC值为0.945,高于单独指标诊断。结论绝经后HHT合并OP患者血清MCP-1、PTX3、GDF15水平升高,且与股骨颈和腰椎(L_(1~4))骨密度、血清骨代谢指标及炎症因子密切相关,三者联合检测对诊断绝经后HHT合并OP患者有较高价值。

着丝粒蛋白H在肾上腺皮质癌的表达及对肾上腺皮质癌细胞活力和迁移的影响

目的:研究着丝粒蛋白H(CENP-H)在肾上腺皮质癌(ACC)的表达,分析其与疾病进展和预后的关系;探索敲减CENP-H表达对ACC细胞活力和迁移的影响。方法:应用GEPIA2数据库分析CENP-H在76例ACC患者和128例健康对照中的mRNA表达,并分析其在不同肿瘤分期的表达及与预后的相关性;运用UALCAN数据库进一步分析CENP-H在ACC不同肿瘤分期的mRNA表达及与预后的相关性;采用免疫组织化学染色检测CENP-H蛋白在ACC和正常肾上腺石蜡切片的表达。构建敲减CENP-H表达的siRNA(siCENP-H),将人ACC细胞系H295R分为siCENP-H组和siNC组;CCK-8实验检测细胞活力的变化,划痕实验检测细胞迁移能力的变化,Western blot实验检测CENP-H、p-ERK1/2、t-ERK1/2、p-P38、t-P38、p-JNK1/2和t-JNK1/2蛋白水平。结果:CENP-H在ACC中表达较健康对照显著升高,且与肿瘤分期和预后显著相关;CENP-H蛋白在ACC组织中的表达水平显著高于正常肾上腺;敲减CENP-H后,H295R细胞活力和迁移能力较对照组显著降低,p-P38和p-JNK1/2蛋白水平显著降低,以p-JNK1/2的降低尤为显著。结论:CENP-H在ACC中高表达;敲减CENP-H表达能抑制ACC细胞活力和迁移,其机制可能与抑制P38和JNK信号通路的活化有关。

基于心肌重塑和心功能标志物NT-proBNP、H-FABP水平探讨黄芪注射液治疗慢性心力衰竭的作用

【目的】基于心肌重塑和心功能标志物N端脑钠肽前体(NT-proBNP)、心型脂肪酸结合蛋白(H-FABP)水平探讨黄芪注射液治疗慢性心力衰竭的作用。【方法】将80例心气虚型慢性心力衰竭患者随机分为对照组和研究组,每组各40例。对照组给予西医常规治疗,研究组在对照组的基础上给予黄芪注射液治疗,疗程为1周。观察2组患者治疗前后Lee氏心衰计分、中医证候积分、明尼苏达心力衰竭生活质量调查问卷(MLHFQ)评分及6 min步行距离的变化情况,检测2组患者治疗前后左室射血分数(LVEF)、每搏输出量(SV)和左室舒张末期内径(LVEDD)等心功能指标以及血清NT-proBNP、H-FABP的水平变化,并评估2组患者的临床疗效。【结果】(1)疗效方面,治疗1周后,研究组的总有效率为90.00%(36/40),对照组为62.50%(25/40),组间比较(χ^(2)检验),研究组的疗效明显优于对照组(P<0.01)。(2)结局指标方面,治疗后,2组患者的Lee氏心衰计分、中医证候积分及MLHFQ评分均较治疗前明显下降(P<0.01),且研究组的下降幅度均明显优于对照组(P<0.01)。(3)6 min步行距离方面,治疗后,2组患者的6 min步行距离均较治疗前明显增加(P<0.05或P<0.01),且研究组的增加幅度明显优于对照组(P<0.05)。(4)实验室指标方面,治疗后,2组患者的血清NT-proBNP、H-FABP水平均较治疗前明显下降(P<0.01),且研究组的下降幅度均明显优于对照组(P<0.01)。(5)心功能指标方面,治疗后,2组患者的LVEF、SV水平均较治疗前明显升高(P<0.01),且研究组的升高幅度均明显优于对照组(P<0.01);而2组患者的LVEDD治疗前后及治疗后组间比较,差异均无统计学意义(P>0.05)。【结论】在西医常规治疗基础上联合黄芪注射液治疗心气虚型慢性心力衰竭患者疗效确切,能有效缓解患者临床症状,改善患者心功能,提高患者生活质量。

Construction of Recombinant Expression Plasmids Containing H and F Protein Genes of Canine Distemper Virus Isolated from a Mink and Their Expression in Prokaryotic Cells

[Objective] This study aimed to construct the recombinant expression plasmids containing H and F protein genes of Canine distemper virus isolated from a mink and to express these two genes in prekaryotic cells as well as to study the reactogenieity of the expressed products. [ Method ] RT-PCR amplification was used to obtain H and F protein genes; TA cloning and subclonlng techniques were used to construct the cloning plasmids（pMD-18T-H and pMD-18T-F） and recombinant expression plasmids（pET28a-H and pET28a-F） ; SDS-PAGE and Western-blotting were adopted to verify whether the target proteins were successfully expressed. [ Result] The recombinant expression plasmids pET28a-H and pET28a-F containing H and F protein genes of Canine distemper virus isolated from a mink were successfully constructed, and both the expressed H and F proteins with respectively relative molecular mass of 31 400 and 38 200 produced positive reac- tion with the CDV standard positive serum. [ Conclusion] The H and F proteins expressed in prokaryotic cells were the same with the natural ones in terms of reac- togenicity, which can be utilized for diagnosis of a CDV＇s infection or for an epidemiological investigation. Meanwhile, they also provide a basis for developing ge- netically engineered subunit vaccines.

益肾排毒方治疗老年糖尿病肾病患者疗效及对24h尿蛋白与β2微球蛋白的影响

目的 探讨益肾排毒方治疗老年糖尿病肾病患者疗效及对24 h尿蛋白、β2微球蛋白的影响。方法 将武汉市中医医院2019年1月—2021年4月收治的老年糖尿病肾病患者101例,随机对照表法随机分为对照组53例和观察组48例。对照组采用降糖、降压、控制蛋白、纠正电解质代谢紊乱、肾性贫血及肾性骨病等对症治疗;在此基础上,观察组采用益肾排毒方治疗,两组均治疗12周。比较两组的临床疗效及不良反应状况,测定白细胞介素-6(interleukin-6,IL-6)、C反应蛋白(C-reactive protein, CRP)、血肌酐(serum creatinine, Scr)、尿素氮(urea nitrogen, BUN)及24 h尿蛋白(24 h urine protein quantification, 24 h Upro)、β2-微球蛋白(β2-microglobulin, β2-MG)。结果 观察组治疗后血清IL-6与CRP低于对照组,差异有统计学意义(P<0.05)。观察组治疗后SCR、BUN及24 h Upro、β2-MG水平低于对照组(P<0.05)。对照组有效率79.25%(42/53)与观察组93.75%(45/48)比较,差异有统计学意义(P<0.05)。对照组总不良率18.87%(10/53)与观察组27.08%(13/48)比较,差异无统计学意义(P>0.05)。结论 益肾排毒方对老年糖尿病肾病患者的疗效显著,可降低患者炎症因子水平,降低24 h Upro、β2-MG,改善肾功能和血管内皮功能,值得推广。

小反刍兽疫病毒H蛋白抗体化学发光免疫分析检测方法的建立

本研究旨在建立小反刍兽疫病毒H蛋白抗体的化学发光免疫分析检测方法。以H蛋白4个反应原性较好的B细胞表位串联后为检测抗原,在确定抗原包被量和血清稀释度,优化血清和酶标二抗反应时间的基础上,用ROC曲线分析确定检测临界值,建立了小反刍兽疫病毒H蛋白抗体化学发光免疫分析检测方法,然后对该方法进行敏感性、特异性和重复性评价。结果显示,建立的小反刍兽疫病毒H蛋白抗体化学发光免疫分析检测方法最佳抗原包被浓度为1.5×10^(-6)μg/孔,待检血清最佳稀释度为1∶100;血清及酶标二抗孵育时间均为10 min,检测方法的临界值为S/P=9.77%,批内及批间变异系数均小于10%;与口蹄疫、羊痘、蓝舌病及山羊传染性胸膜肺炎阳性血清不发生交叉反应;用建立的化学发光法和市售小反刍兽疫抗体cELISA试剂盒平行检测247份田间血清,两者相对符合率为94.33%,但化学发光法敏感性更高。研究结果表明,建立的小反刍兽疫病毒H蛋白抗体化学发光免疫分析方法灵敏、特异、稳定,可用于田间血清样品小反刍兽疫抗体检测。

免疫检查点HHLA2在子宫内膜癌中的表达及临床意义

目的:探索免疫检查点人内源性逆转录病毒-H长末端重复关联蛋白2(HHLA2)在子宫内膜癌中的表达及临床意义。方法:回顾性收集192例子宫内膜癌患者的组织标本及临床病理资料,利用Sanger测序检测POLE突变,免疫组化染色检测错配修复蛋白及p53蛋白表达。将子宫内膜癌划分为POLE突变型、错配修复缺陷型(MMRd)、p53突变型及无特定分子特征(NSMP)型,免疫组化染色检测HHLA2蛋白表达,利用卡方检验分析HHLA2蛋白与临床病理特征及分子分型的关系。采用Kaplan-Meier法和Cox比例风险回归模型分析影响子宫内膜癌患者预后的相关因素。结果:HHLA2蛋白在子宫内膜癌中的阳性表达率为69.8%(134/192)。子宫内膜样癌、中-高分化肿瘤、I期、浅肌层浸润、无淋巴脉管间隙浸润及分子分型为NSMP型的肿瘤中HHLA2蛋白表达率较高。整个研究队列及分子分型为NSMP型的亚组患者中,HHLA2蛋白表达阳性患者的无疾病复发生存率高于HHLA2蛋白表达阴性患者(P<0.05)。多因素生存分析显示,分子分型是子宫内膜癌患者的预后因素(P<0.05)。结论:HHLA2是NSMP型子宫内膜癌的预后因素和潜在治疗靶点。

Overexpression of centromere protein H is significantly associated with breast cancer progression and overall patient survival

Breast cancer is one of the leading causes of cancer death worldwide.This study aimed to analyze the expression of centromere protein H(CENP-H) in breast cancer and to correlate it with clinicopathologic data,including patient survival.Using reverse transcription-polymerase chain reaction and Western blotting to detect the expression of CENP-H in normal mammary epithelial cells,immortalized mammary epithelial cell lines,and breast cancer cell lines,we observed that the mRNA and protein levels of CENP-H were higher in breast cancer cell lines and in immortalized mammary epithelial cells than in normal mammary epithelial cells.We next examined CENP-H expression in 307 paraffin-embedded archived samples of clinicopathologically characterized breast cancer using immunohistochemistry,and detected high CENP-H expression in 134(43.6%) samples.Statistical analysis showed that CENP-H expression was related with clinical stage(P = 0.001),T classification(P = 0.032),N classification(P = 0.018),and Ki-67(P < 0.001).Patients with high CENP-H expression had short overall survival.Multivariate analysis showed that CENP-H expression was an independent prognostic indicator for patient survival.Our results suggest that CENP-H protein is a valuable marker of breast cancer progression and prognosis.

Construction of an oral recombinant DNA vaccine from H pylori neutrophil activating protein and its immunogenicity

AIM: To construct a live attenuated Salmonella ty- phimurium (S. typhimurium) strain harboring the H pylori neutrophil activating protein (HP-NAP) gene as an oral recombinant DNA vaccine, and to evaluate its immuno- genicity. METHODS: By genetic engineering methods, the ge- nomic DNA of H pylori was extracted as a template. The total length of the HP-NAP gene was amplified by poly- merase chain reaction (PCR) and cloned into pBT vector for sequencing and BLAST analysis, then subcloned into a eukaryotic expression vector pIRES followed by PCR identification and restriction enzyme digestion. The iden- tified recombinant plasmid pIRES-NAP was transfected into COS-7 cells for target fusion protein expression, and its antigenicity was detected by Western blotting. Then the recombinant plasmid was transformed into a live attenuated S. typhimurium strain SL7207 as an oral vac- cine strain, and its immunogenicity was evaluated with animal experiments. RESULTS: A 435 bp product was cloned using high homology with HP-NAP gene in GenBank (more than 98%). With identification by PCR and restriction enzyme digestion, a recombinant eukaryotic expression plasmid pIRES-NAP containing the HP-NAP gene of H pylori was successfully constructed. The expressed target protein had a specific reaction with H pylorii whole cell antibody and showed a single strip result detected by Western blotting. Oral immunization of mice with recombinant DNA vaccine strain SL7207 (pIRES-NAP) also induced a specific immune response. CONCLUSION: The successful construction of HP-NAP oral DNA vaccine with good immunogenicity may help to further investigate its immunoprotection effects and de- velop vaccine against H pylori infection.

Dengue virus non-structural 1 protein interacts with heterogeneous nuclear ribonucleoprotein H in human monocytic cells

Objective: To study protein-protein interaction between heterogeneous nuclear ribonucleoprotein H(hn RNP H) and Dengue virus(DENV) proteins. Methods: DENV proteins were screened against the host hn RNP H protein, in order to identify the host-viral protein-protein interactions in DENV infected THP-1 cells by co-immunoprecipitation. The co-localization of the interacting proteins was further confirmed by immunofluorescence microscopy. Results: The host protein hn RNP H was found to interact with DENV nonstructural 1 protein and help the virus to multiply in the cell. Conclusions: The non-structural 1 glycoprotein is a key modulator of host immune response and is also involved in viral replication. Therefore, disruption of this key interaction between hn RNP H and DENV nonstructural 1 could be an important therapeutic strategy for management of DENV infection.

H pylori infection and systemic antibodies to CagA and heat shock protein 60 in patients with coronary heart disease

AIM: To determine the overall prevalence of H pylori and CagA positive H pylori infection and the prevalence of other bacterial and viral causes of chronic infection in patients with coronary heart disease (CHD), and the potential role of anti-heat-shock protein 60 (Hsp60) anti- body response to these proteins in increasing the risk of CHD development. METHODS: Eighty patients with CHD and 160 controls were employed. We also compared the levels of anti- heat-shock protein 60 (Hsp60) antibodies in the two groups. The H pylori infection and the CagA status were determined serologically, using commercially available enzyme-linked immunosorbent assays (ELISA), and a Western blotting method developed in our laboratory. Systemic antibodies to Hsp60 were determined by a sandwich ELISA, using a polyclonal antibody to Hsp60 to sensitise polystyrene plates and a commercially available human Hsp60 as an antigen. RESULTS: The overall prevalence of H pylori infec- tion was 78.7% (n = 63) in patients and 76.2% (n = 122) in controls (P = 0.07). Patients infected by CagA- positive (CagA+) H pylori strains were 71.4% (n = 45) vs 52.4% of infected controls (P = 0.030, OR = 2.27). Sys-temic levels of IgG to Hsp60 were increased in H pylori- negative patients compared with uninfected controls (P < 0.001) and CagA-positive infected patients compared with CagA-positive infected controls (P = 0.007). CONCLUSION: CagA positive H pylori infection may concur to the development of CHD; high levels of anti- Hsp60 antibodies may constitute a marker and/or a con- comitant pathogenic factor of the disease.

anti-EGFR scFv-FTH1/AP1-FTH1纳米粒子的构建及其治疗哮喘小鼠的应用

采用基因工程方法分别将抗表皮生长因子受体(EGFR)的单链抗体(anti-EGFR scFv)和靶向白介素-4-受体(IL-4R)的AP1短肽分别修饰到铁蛋白重链亚基(FTH1)的N端,再通过蛋白表达、体外混合复性后成功制备了anti-EGFR scFv-FTH1/AP1-FTH1双靶向纳米粒子。结果表明:该纳米粒子能正确组装成铁蛋白的特征笼状结构,粒径为(13.2±1.3)nm。在哮喘小鼠模型中发现该纳米粒子能有效抑制哮喘多种症状,且在缓解气道高反应方面的效果优于单靶向anti-EGFR scFv-FTH1/FTH1纳米粒子。

血清蛋白电泳联合24 h尿蛋白定量在糖尿病肾病患者诊断中的应用价值分析

目的 通过比较糖尿病肾病(DN)患者与健康人群的血清蛋白电泳与24 h尿蛋白定量水平,分析其临床诊断价值。方法 选取50例明确诊断的DN患者作为观察组,另选取同期50例健康体检者作为对照组。两组均进行血清蛋白电泳及24 h尿蛋白定量检测。比较两组的血清蛋白电泳[白蛋白(ALB)、β_(1)-球蛋白、β_(2)-球蛋白、α_(1)-球蛋白、α_(2)-球蛋白、γ-球蛋白]水平及24 h尿蛋白定量水平、阳性率。结果 观察组的α_(1)-球蛋白(6.0±1.9)%、α_(2)-球蛋白(13.1±3.7)%、β_(2)-球蛋白(6.5±1.5)%均显著高于对照组的(3.9±0.6)%、(9.3±2.0)%、(4.6±1.0)%,ALB(46.1±10.7)%显著低于对照组的(60.0±2.2)%,差异有统计学意义(P<0.05);两组β_(1)-球蛋白与γ-球蛋白比较,差异无统计学意义(P>0.05)。结论 血清蛋白成分与24 h尿蛋白定量的改变能够辅助诊断DN,可以有效评估DN患者的治疗效果,并对预测远期效果意义重大,而且此方法简单可靠,可以在临床中广泛应用。

H pylori stimulates proliferation of gastric cancer cells through activating mitogen-activated protein kinase cascade

AIM: To explore the mechanism by which H pylori causes activation of gastric epithelial cells. METHODS: A VacA (+) and CagA (+) standard H pylori line NCTC 11637 and a human gastric adenocarcinoma derived gastric epithelial cell line BGC-823 were applied in the study. MTT assay and 3H-TdR incorporation test were used to detect the proliferation of BGC-823 cells and Western blotting was used to detect the activity and existence of related proteins. RESULTS: Incubation with H pylori extract increased the proliferation of gastric epithelial cells, reflected by both live cell number and DNA synthesis rate. The activity of extracellular signal-regulated protein kinase (ERK) signal transduction cascade increased within 20 min after in- cubation with H pylori extract and appeared to be a sus- tained event. MAPK/ERK kinase (MEK) inhibitor PD98059 abolished the action of H pylori extract on both ERK activity and cell proliferation. Incubation with H pylori extract increased c-Fos expression and SRE-dependent gene expression. H pylori extract caused phosphorylation of several proteins including a protein with molecular size of 97.4 kDa and tyrosine kinase inhibitor genistein inhibited the activation of ERK and the proliferation of cells caused by H pylori extract. CONCLUSION: Biologically active elements in H pylori extract cause proliferation of gastric epithelial cells through activating tyrosine kinase and ERK signal trans- duction cascade.

湿热处理对花生球蛋白Ara h 3抗原性的影响

为探究湿热处理对花生球蛋白Ara h 3抗原性的影响,采用硫酸铵分级沉淀法从花生脱脂粉中分离纯化出Ara h 3,进行不同温度、时间和质量浓度的湿热处理,通过正交试验确定最优湿热处理条件,采用间接ELISA法检测湿热处理后Ara h 3抗原性变化,测定外源荧光、游离巯基含量以及圆二色光谱,从蛋白结构的变化解释湿热处理对Ara h 3抗原性的影响。研究表明:湿热处理能够降低Ara h 3抗原性,且在质量浓度10 mg/mL、90℃处理30 min时,抗原性降至最低;湿热处理后的Ara h 3荧光强度、表面疏水性增强,三级结构发生变化;游离巯基含量增加、空间结构发生改变;α-螺旋和β-折叠含量降低,无规则卷曲含量增加,二级结构发生变化。湿热处理后Ara h 3空间结构发生改变,部分抗原表位遭到破坏或被包埋,抗原性降低。

拉贝洛尔对重度子痫前期患者血压、24 h尿蛋白水平及分娩结局的影响

目的分析重度子痫前期患者接受拉贝洛尔治疗对其血压、24 h尿蛋白水平和分娩结局的影响。方法60例重度子痫前期患者,根据入院顺序分为对照组和研究组,每组30例。两组患者均给予常规治疗,在此基础上,对照组患者配合硝苯地平治疗,研究组患者配合拉贝洛尔治疗。比较两组患者血压、24 h尿蛋白水平和不良分娩结局情况。结果研究组患者收缩压、舒张压及24 h尿蛋白水平分别为(139.27±13.68)mm Hg(1 mm Hg=0.133 kPa)、(84.02±6.94)mm Hg、(2670.66±42.78)mg,明显低于对照组的(150.33±13.75)mm Hg、(90.32±4.36)mm Hg、(2960.33±41.48)mg,差异有统计学意义(P<0.05)。研究组患者治疗后的不良分娩结局发生率23.3%(7/30)明显低于对照组的70.0%(21/30),差异有统计学意义(P<0.05)。结论拉贝洛尔治疗重度子痫前期患者的临床疗效确切,能够帮助控制患者血压及24 h尿蛋白水平,改善患者不良分娩结局,应该给予大力的推广与应用。