腹腔热化疗对胃肠道恶性肿瘤患者T淋巴细胞亚群及sIL-2R水平的影响

为评估腹腔灌注热化疗（ＩＰＨＣ）对胃肠道恶性肿瘤患者免疫功能的影响,对３２例胃肠道恶性肿瘤患者施行术后腹腔灌注热化疗,测定其外周血Ｔ淋巴细胞亚群（ＴＬＳ）和血清白细胞介素Ⅱ受体（ｓＩＬ２Ｒ）水平,与２０例健康对照者进行比较。结果：行ＩＰＨＣ前,ＣＤ＋３、ＣＤ＋４、ＣＤ＋８、ＣＤ＋４／ＣＤ＋８较对照组降低,ｓＩＬ２Ｒ升高;行ＩＰＨＣ后,ＣＤ＋３、ＣＤ＋４、ＣＤ＋８、ＣＤ＋４／ＣＤ＋８明显下降（Ｐ＜０．０１）,ｓＩＬ２Ｒ降低（Ｐ＜０．０１）。结论：腹腔灌注热化疗能明显提高患者免疫功能。     

表达白细胞介素10的系膜细胞基因转移载体的建立

目的为特异性地将病毒性白细胞介素１０（ｖＩＬ１０）基因转移到肾脏提供一种手段。方法应用逆转录病毒载体介导的基因转移技术将外源性基因ｖＩＬ１０转移到大鼠肾脏系膜细胞,应用ＲＴＰＣＲ、ＥＬＩＳＡ方法检测其表达,并通过对混合淋巴细胞反应（ＭＬＲ）的影响对其活性进行分析。结果ｖＩＬ１０在转染的系膜细胞表达２００００ｐｇ·（１０６细胞）－１／２４ｈ,应用３ＨＴｄＲ掺入法检测ｖＩＬ１０对ＭＬＲ的影响,实验组明显低于对照组（Ｐ＜００５）。结论外源性基因在系膜细胞有效地转录和表达,并对ＭＬＲ有抑制作用。     

狼疮肾炎患者血与尿白细胞介素-8测定的临床意义

目的　探讨血清和尿中白细胞介素８（ Ｉ Ｌ８） 水平变化在系统性红斑狼疮（ Ｓ Ｌ Ｅ） 及狼疮肾炎（ Ｌ Ｎ） 中的临床意义。方法　采用 Ｅ Ｌ Ｉ Ｓ Ａ 法对４７ 例 Ｓ Ｌ Ｅ 患者（ 含３７ 例 Ｌ Ｎ） 、１３ 例原发性慢性肾小球肾炎和１４ 例正常对照进行血清和尿中 Ｉ Ｌ８ 水平检测,比较分析其与 Ｓ Ｌ Ｅ 活动指数（ Ｓ Ｌ Ｅ Ｄ Ａ Ｉ） 、肾脏损害的相关关系。结果　 Ｓ Ｌ Ｅ 患者血清 Ｉ Ｌ８ 水平活动期为５５３ ｐｇ／ｍｌ（ 中位数,下同） 静止期为２６３８ ｐｇ／ｍｌ,明显高于正常对照组２０５４ ｐｇ／ｍｌ（ Ｐ ＜ ０００１） ,并且活动期高于静止期（ Ｐ ＜ ００５） ,血清 Ｉ Ｌ８ 水平与 Ｓ Ｌ Ｅ Ｄ Ａ Ｉ正相关（ ｒ ＝ ０６００３ , Ｐ ＜ ００５） ; Ｓ Ｌ Ｅ 患者尿 Ｉ Ｌ８ 显著高于正常对照组２０６７ｐｇ／ｍｌ（ Ｐ＜ ００５） ,且 Ｌ Ｎ 组２５２９ ｐｇ／ｍｌ 明显高于 Ｓ Ｌ Ｅ 无肾炎组为２１１ ｐｇ／ｍｌ 及原发性慢性肾小球肾炎组１９４１ ｐｇ／ｍｌ（ Ｐ 值均＜ ００５） ;肾脏组织病理狼疮活动指数（ Ａ Ｉ） 高者其血清、尿 Ｉ Ｌ８ 水平显著高于 Ａ Ｉ低者（ Ｐ ＜ ００５） 。结论　血清 Ｉ Ｌ     

Comparison of the nuclear matrix protein 22 with voided urine cytology in the diagnosis of transitional cell carcinoma of the bladder

Several urinary markers for transitional cell carcinoma have been investigated, including urine cytology, bladder tumor antigen, autocrine motility factor receptor and fibrin degradation products. Unfortunately, they have poor overall sensitivity. The United States Food and Drug Administration have recently approved nuclear matrix protein (NMP 22) for the detection of occult or rapidly recurring disease after transurethral resection of bladder tumor. The objective of the current study was to assess the sensitivity of NMP 22 for the detection of bladder carcinoma, as well as to correlate the NMP 22 values with multiplicity of tumor, tumor size, configuration, stage and grade respectively. A total of 78 patients (38 with bladder cancer) provided a urine sample which was divided into appropriate aliquots for each of urine cytology and NMP 22. Comparative results demonstrate a clear superiority of NMP 22 in bladder cancer detection (52.6% vs 31.6% sensitivity), while specificity was in favor of urine cytology (100% vs 82.5%). For superficial tumors, sensitivity was 78.5% for NMP 22 and 41.6% for cytology and for invasive cancers, sensitivity was 90% for NMP 22 and 60% for cytology. Urinary NMP 22 levels were significantly correlated with tumor grade and were significantly higher in large tumors than small tumors. NMP 22 test results showed sufficient sensitivity in comparison with urine cytology for the detection of transitional cell carcinoma. However, we do not think that it is a useful tool as a substitute for endoscopic examination for the detection and surveillance in bladder cancer.     

Decreased collagen-degrading activity could be a marker of prolonged mesangial matrix expansion

Background Mesangial matrix expansion is caused by the overproduction and/or the impaired proteolytic degradation of the extracellular matrix. However, the relative contribution of these changes to the development of prolonged mesangial matrix expansion is still poorly understood. We aimed to elucidate the relative role of the matrix metalloproteinase (MMP)/tissue inhibitors of metalloproteinases (TIMPs) system in the development of prolonged mesangial matrix expansion.Methods We prepared two rat models, showing reversible or prolonged mesangial matrix expansion, induced by a single injection or two consecutive injections of anti-Thy-1.1 monoclonal antibody 1-22-3, respectively. We analyzed the glomerular expression of type I and type IV collagens; MMP-2, -9, and -13; membrane type 1-MMP (MT1-MMP); TIMP-1; and urinary type I collagen-degrading activity in both models.Results There were no differences in glomerular mRNA levels of type I and type IV collagens between the reversible and the prolonged models. MMP-9 mRNA expression and protein level was lower in the prolonged model than in the reversible one, whereas there were no differences in mRNA levels of MMP-2, -13, MT1-MMP, or TIMP-1 between the two models. Urinary type I collagen-degrading activity in the prolonged model was lower than that in the reversible one. Furthermore, there was a significant correlation between the mesangial matrix expansion and urinary type I collagen-degrading activity.Conclusions Impaired expression of MMP-9 may contribute to the development of prolonged mesangial matrix expansion. Analysis of urinary type I collagen-degrading activity may provide additional diagnostic information in mesangial proliferative glomerulonephritis.     

Changes of interleukin-1 β, tumor necrosis factor α and interleukin-6 in brain and plasma after brain injury in rats

Objective: To study the changes of interleukin-1 β(IL-1β), tumor necrosis factor α (TNFα) and interleukin-6 (IL-6) levels in brain and plasma after brain injury and to assess the relationship between the cytokine levels and injury severity in rats. Methods. A total of 51 male Wistar rats, weighing 280-340g, were anesthetized with chloral hydrate(400 mg/kg body weight) through intraperitoneal injection and fixed on a stereotaxic instrument. Severe brain injury was created in 16 rats (severe injury group) and moderate brain injury in 18 rats (moderate injury group) by a fluid percussion model, and cytokine levels of IL-1β, TNFα and IL-6 were measured with biological assay. And sham operation was made on the other 17 rats (control group). Results: In the control group, the levels of IL-1β,TNFα and IL-6 were hardly detected in the cortex of the rats, but in the ipsilateral cortex of the rats in both injury groups, they increased obviously at 8 hours after injury.The increasing degree of these cytokines had no significant difference between the two injury groups. The levels of IL-6 in the plasma of all the rats increased slightly, whereas the levels of IL-1β and TNFα were undetectable. Conc|usions: The increase of IL-1β, TNFα and IL-6 levels is closely related to brain injury. The increased cytokine levels in the central nervous system are not parallel to those in the peripheral blood. It suggests that inflammatory cytokines play important roles in the secondary neural damage after brain injury.     

丙戊酸对大鼠急性脊髓损伤后神经功能恢复的影响及作用机制

目的:探讨丙戊酸(VPA)对大鼠脊髓损伤(SCI)后运动功能恢复的影响及作用机制。方法:60只雄性SD大鼠随机均分为3组:假手术组(C组)、损伤组(SCI组)和丙戊酸保护组(VPA组)。SCI组和VPA组采用改良Allen法制作大鼠T10 SCI模型。VPA组术后即刻及其后每12h皮下注射VPA 300mg/kg,至取材;C组和SCI组在相应时间点注射等体积的生理盐水。伤后6h,每组取5只大鼠处死取材,其余大鼠在伤后24h、48h和72h每组取5只先行后肢运动功能BBB评分,随后处死取材。切片后分别行HE染色观察脊髓组织病理变化,免疫荧光双标法在激光共聚焦显微镜下观察核因子κB(NF-κB)途径的激活状态,免疫组化法检测白介素1β(IL-1β)的表达。结果:C组大鼠各时间点BBB评分均为21分,VPA组和SCI组各时间点的评分均低于C组(P<0.05),但VPA组各时间点的评分均高于同时间点SCI组,在伤后48h和72h两组差异有显著性(P<0.05)。病理检查显示C组脊髓组织形态正常,VPA组和SCI组伤后6h损伤中央区即可见明显出血灶,灰质中神经元肿胀坏死,白质中神经纤维肿胀;伤后24h、48h出血灶界限更明显,并可见空洞形成和炎症细胞浸润;伤后72h上述病理变化仍明显;VPA组各时间点的病理变化与SCI组相似,但炎症细胞浸润减少。C组偶见或未见NF-κB核阳性细胞和IL-1β表达,与C组相比,SCI组和VPA组NF-κB核阳性细胞百分比和IL-1β表达量从伤后6h即显著性增高,24h达高峰,以后逐渐减少,72h仍显著性高于C组(P<0.05);VPA组各时间点NF-κB核阳性细胞百分比和IL-1β表达量均低于同时间点SCI组(P<0.05)。结论:VPA可促进大鼠SCI后运动神经功能恢复,其机制可能与抑制炎症反应有关。     

稳定沉默Rictor表达的前列腺癌CWR22RV1的构建

目的 应用RNA沉默技术下调前列腺癌CWR22RV1细胞株的Rictor基因表达,并筛选具有稳定转染Rictor-shRNA的细胞株,为进一步研究Rictor在前列腺癌中的作用打下基础.方法 针对Rictor基因设计合成siRNA序列;鉴定、测序并转染293T细胞观察基因表达情况;构建Rictor-shRNA慢病毒载体;进行Rictor-shRNA慢病毒包装及滴度测定;筛选稳定表达Rictor-shRNA的前列腺癌CWR22RV1细胞株;Western blot检测稳定转染前列腺癌CWR22RV1细胞株Rictor的表达水平.结果 Rictor-shRNA慢病毒成功包装后转染前列腺癌CWR22RV1细胞,进行嘌呤霉素筛选,获得沉默Rictor基因的前列腺癌CWR22RV1细胞株,Western blot检测显示该细胞株Rictor水平明显降低(P＜0.01).结论 成功构建了稳定沉默Rictor表达的前列腺癌CWR22RV1细胞株,为后续的实验了打下坚实的基础.     

MiR-30a-5p通过泛素水解酶22抑制人宫颈癌细胞上皮-间质转化功能

【目的】研究microRNA-30a-5p（miR-30a-5p）对人宫颈癌Hela细胞上皮-间质转化功能的影响及其相关机制。【方法】宫颈癌Hela细胞株分别转染目的mir的模拟物和阴性对照模拟物,分别以30a-5p组、NC组命名并标记细胞。同时,以未经过处理的Hela细胞作为对照（Control组）。分别用逆转录-聚合酶链反应法检测各组宫颈癌细胞的miR-30a-5p含量。Transwell实验检测3组细胞迁移能力和侵袭能力。Western-blot法检测3组细胞神经-钙粘素（N-cadherin）、α-连环蛋白（α-Catenin）和泛素水解酶22（USP22）表达水平。运用生物信息学方法预测miR-30a-5p的靶基因。采用Western blot法检测USP22过表达对miR-30a-5p抑制EMT的拮抗作用。双荧光素酶实验检测miR-30a-5p与USP22的关系。建立皮下移植瘤模型观察miR-30a-5p的体内作用。【结果】30a-5p组宫颈癌细胞miR-30a-5p的表达水平明显上调,表达水平为Control组的853.82（862.26~843.11）倍（P＜0.01）。30a-5p组侵袭细胞数量8.17（8.32~8.03）明显低于Control组（P＜0.01）。30a-5p组细胞N-cadherin蛋白的细胞内含量明显下降,α-Catenin蛋白的细胞内含量明显上升,USP22蛋白表达量明显降低。合并USP22过表达处理的30a-5p组宫颈癌细胞中N-cadherin蛋白表达量明显升高,α-Catenin蛋白表达量明显降低。双荧光素酶检验结果显示USP22为miR-30a-5p的下游靶基因（P＜0.01）。30a-5p组皮下移植瘤明显小于Control组（P＜0.01）。与Control组肿瘤组织相比,30a-5p组肿瘤组织miR-30a-5p的相对含量升高,USP22蛋白含量降低,N-cadherin蛋白的含量降低,α-Catenin蛋白含量升高。【结论】miR-30a-5p在宫颈癌Hela细胞中,可能通过靶向识别下游靶基因USP22,进而抑制其翻译。最终实现对宫颈癌细胞EMT过程的抑制。     

A preliminary study of cytokine gene polymorphism effects on Saudi patients with colorectal cancer

Objectives:To determine the possible associations of polymorphisms in interleukin (IL)-8 (rs4073 T/A), IL-10 (rs1800896 A/G), IL-22 (rs1179251 C/G and rs2227485 C/T), IL-27 (rs17855750 T/G), and transforming growth factor beta 1 (TGFß1) (rs1800469 C/T) with colorectal cancer (CRC) susceptibility in Saudi patients.Methods:The case-control study was carried out between July 2019 and January 2020 in King Khaled University Hospital, Riyadh, Saudi Arabia. A total of 70 patients with CRC and 70 healthy controls were included in the study. Single nucleotide polymorphisms of promoter regions were determined using TaqMan genotyping assays.Results:A statistically significant reduction in CRC risk was identified for carriers of the IL-10 (rs1800896 A/G) AG genotype, IL-22 (rs1179251 C/G) G allele, IL-27 (rs17855750 T/G) G allele and TGFß1 (rs1800469 C/T) CT and TT genotype. While IL-10 (rs1800896 A/G) AA genotype and TGFß1 (rs1800469 C/T) CC genotype were significantly associated with increased susceptibility to CRC. No significant associations were identified between the cytokine polymorphisms of IL-8 (rs4073 T/A) and IL-22 (rs2227485 C/T), and CRC risk.Conclusion:Our findings indicate a significant impact of IL-10 (rs1800896 A/G), IL-22 (rs1179251 C/G), IL-27 (rs17855750 T/G) and TGF-ß1 (rs1800469 C/T) polymorphisms on risk of CRC; while the IL-8 (rs4073 T/A) and IL-22 (rs2227485 C/T) and polymorphisms were not associated with CRC risk.