Expression of B7 costimulation molecules by colorectal cancer cells reduces tumorigenicity and induces anti-tumor immunity

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Cloning and expression of MXR7 gene in human HCC tissue

AIM To clone and identify the whole cDNA ofMXR7 gene and to find out its expression inhuman HCC,and normal tissues.METHODS The DNA primers were designed andsynthesized according to the whole cDNAsequence of MXR? gene.The cDNA of humanHCC was taken as the template while the cDNA ofMXR7 gene was synthesized by polymerasechain reaction（PCR）.Recombinant DNAconforming to reading frame was constructed byconnecting purified PCR product of the cDNA ofMXR? gene with expression vector pGEX-5X-1 offusion protein.The plasmid MXRT/pGEX-5X-1was identified by sequencing.Using 32p labeledMXR? cDNA as probe,MXR7 mRNA expressionwas detected by Northern blot analysis in 12different human normal tissues,7 preoperativelyuntreated non-liver tumor tissues,30preoperatively untreated HCC,theparacancerous liver tissues and 12 normal livertissues samples.RESULTS Restriction enzyme and sequenceanalysis confirmed that the insertion sequence invector pGEX-5X-1 was the same as the cDNAsequence of MXR7 gene.Northern blot analysisshowed no expression of MXR? mRNA in 12 kindsof normal human tissues including liver,7 tumortissues in other sites and 12 normal livertissues,the frequencies of MXR7 mRNA expression in HCC and paracancerous livertissues were 76.6% and 13.3%,respectively.The frequency of MXR7 mRNA expression in HCCwithout elevation of serum AFP and in HCC【5cm was 90%（9/10）and 83.3%（5/6）,respectively.CONCLUSION MXR7 mRNA is highly expressedin human HCC,which is specific and occurs at anearly stage of HCC,suggesting MXR7 mRNA canbe a tumor biomarker for HCC.The detection ofMXR7 mRNA expression in the biopsied livertissue is helpful in discovering early subclinicalliver cancer in those with negative serum AFP.

Expression and significance of MMP-7,c-Jun and c-Fos in rats skin photoaging

Objective:To investigate the expression and significance of the MMP-7,c-Jun and c-Fos in rat photoaging skin.Methods:A total of 45 SD rats were randomly divided into control group,model group,natural recovery group,physiological saline injection group and dermal pluripotent stem cells transplantation(DMSCs group),model group,natural recovery group,physiological saline injection group.DMSCs were treated with UV lamp irradiation to establish light aging skin model.Rats were then sacrificed after model prepared,no treatment was processed in the natural recovery group.Saline injections was adopted in saline group,DESCs group was treated with DESCs transplantation.Rats were sacrificed after 4 weeks.The expression of MMP-7,c-Jun and c-Kos were detected using the immunohistocheniical metluxl.Results:In model group,MMP 7positive expression was higher than that in the other 4 groups,but without statistically difference(P>0.05);c-Jun,c-Fos expression were higher than that in the control group and DESCs group(P<0.05),there was no significant difference comparing natural recovery group with physiological saline injection group(P>0.05).Conclusions:MMP-7,c-Jun and c-Fos can be used as diagnosis indicators in the early stage of light aging,and they jointly participate in its development.DMSCs transplants is effective in treating light aging skin.

Cloning and expression of HLA-B7 gene

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基于心功能及IGFBP7、sST2、CGRP、ET分析沙库巴曲缬沙坦在治疗冠心病合并慢性心力衰竭中的应用效果

目的 分析冠心病(CHD)合并慢性心力衰竭(CHF)患者应用沙库巴曲缬沙坦治疗的效果。方法 选择2020年1月至2023年1月邯郸市第四医院收治的86例CHD合并CHF患者,以随机数字表法将其分为对照组和试验组各43例。两组CHD治疗均应用硝酸酯类、他汀类及抗血小板药物,对照组CHF治疗应用坎地沙坦酯片、醛固酮受体拮抗剂及β受体阻滞剂,试验组治疗则将对照组中的坎地沙坦酯片替换为沙库巴曲缬沙坦钠片。比较两组疗效、不良反应、心功能指标[左室短轴缩短率(LVFS)、左室射血分数(LVEF)、6min步行距离(6 MWD)]、心室重构指标[Ⅲ型胶原前肽(PⅢP)、层粘蛋白(LN)、基质金属蛋白酶-9(MMP-9)]、心肌损伤和血管内皮功能相关指标[胰岛素样生长因子结合蛋白7(IGFBP7)、可溶性生长刺激表达基因2(sST2)、降钙素基因相关肽(CGRP)、内皮素(ET)]。结果与对照组比,试验组治疗3个月后的总有效率更高,差异有统计学意义(P<0.05)。两组治疗3个月后的LVFS、LVEF、6 MWD、IGFBP7、CGRP与治疗前比升高,且试验组与对照组比更高,差异有统计学意义(P<0.05);PⅢP、LN、MMP-9、sST2、ET降低,试验组与对照组比更低,差异有统计学意义(P<0.05)。两组不良反应总发生率对比差异无统计学意义(P>0.05)。结论 沙库巴曲缬沙坦可有效调节CHD合并CHF患者IGFBP7、sST2、CGRP、ET,改善血管内皮功能、心肌损伤、心室重构及心功能,进而可提高疗效,且具有良好的安全性。

Effects of intra-gastric beta-casomorphin-7 on somatostatin and gastrin gene expression in rat gastric mucosa

瞄准：调查实际上在老鼠胃粘膜的胃的 somatostatin 和促胃液素送信人 RNA 的规定上的 beta-casomorphin-7。方法：Somatostatin 和促胃液素 mRNA 被 RT-PCR 和原位杂交(ISH ) 在 24 只老鼠确定。老鼠被划分成三个治疗组：基础饮食 + 生理盐水(n = 8 ) ，基础饮食 + beta-casomorphin-7 (7.5 乘 10-7 摩尔)(n = 8 ) ，并且基础饮食 +poly-Gly-7 (多样地与 7.5 包含 N 的相等的摩尔 10-7 摩尔 beta-casomorphin-7 )(n = 8 ) 。在为 30 天的口服以后，老鼠被 exsanguinations 打死。结果：在为 30 d 的 beta-casomorphin-7 的 intra 胃的管理以后，促胃液素 mRNA 增加了 52.8%(P 【 0.05， n = 8 ) ，并且 somatostatin mRNA 层次与控制相比在 30.7% 减少了(P 【 0.01， n = 8 ) 。尽管促胃液素 mRNA 表示被 35.6% 与控制组对照提高，在二基因的表达式的有效差量都没在 poly-Gly-treated 组被观察(P = 0.15， n = 8 ) 。一个 casomorphin 答案的长期的口服显著地减少了 D 房间的平灰色，但是在窦和宫底两个都降低 D 房间的数字。有趣地， G 房间的数字在窦和宫底增加了，但是它的平均密度仅仅在窦被扩充。结论：Beta-casomorphin-7 能够从 G 和 D 的规章的肽的 modulating 基因表示。从原位杂交研究的数据显示 beta-casomorphin-7 借助于 somatostatin 的 paracrine 行动间接地影响促胃液素基因表示，并且取决于它的内在的分子的函数。

Cloning and Characterization of Porcine TSARG7 Gene and Analysis of Its Tissue-Specific Expression

TSARG7 is a novel member of the acyltransferase family since its sequence possesses the highly conserved phosphate acyltransferase （PlsC） domain existing in all acyltransferase-like proteins. The porcine TSARG7 had been identified by cloning in silico but had not been confirmed experimentally. The full-length mRNA of porcine TSARG7 gene was sequenced and two splice variants were discovered. The full-length cDNA of TSARG7 variant 1 was 2 513 bp and variant 2 was 2 634 bp. The putative porcine TSARG7 proteins, which were located in the cytoplasm, encoded 458 and 456 amino acids, respectively. Real-time PCR analysis showed that TSARG7 gene was expressed in various tissues, but at different levels. The expression levels of this gene were higher in the skeletal muscle, heart, and testis than that in other tissues, suggesting that the TSARG7 gene played a role in procine skeletal muscle, heart, and testis functions.

Expression of the B7 - related molecule B7 - H1 by glioma cells: a potential mechanism of immune paralysis

Human glioblastoma is a highly lethal tumor that is known for its immune inhibitory capabilities.B7-homologue l(B7-H 1),a recently identified homologue of B7.1/2(CD80/86),has been described to exert costimulatoryand immune regulatory functions.We investigated the expression and the functional activity of B7-H 1 in humanglioma cells in vitro and in vivo.Although lacking B7.1/2(CD80/86),all 12 glioma cel1 1ines constitutivelyexpressed B7-H1 mRNA and protein.Exposure to IFN-gamma strongly enhanced B7-H 1 expression.Im-

TIMP-2·IGFBP-7对成例肾病综合征急性肾损伤发生发展的预测价值

目的 明确[TIMP-2]×[IGFBP-7]对成例肾病综合征急性肾损伤发生的预测价值。方法 选取2020年12月—2022年12月济南市第八人民医院收治的102例肾病综合征患者为研究对象,根据急性肾损伤(acute kidney injury, AKI)的发生情况分为AKI组(n=29)及非AKI组(n=73)。对比两组患者的TIMP-2、IGFBP-7、[TIMP-2]×[IGFBP-7]水平,并评估其预测价值。结果 AKI组入院时的TIMP-2、IGFBP-7、[TIMP-2]×[IGFBP-7]水平分别为(3.34±0.57)ng/mL、(159.48±28.95)ng/mL、(543.54±168.17)(ng/mL)2均高于非AKI组,差异有统计学意义(t=11.507、9.184、12.132,P<0.001)。[TIMP-2]×[IGFBP-7]预测AKI的AUC、95%CI、P值、约登指数、敏感度及阴性预测值均高于单一TIMP-2预测和IGFBP-7预测,差异有统计学意义(P<0.001)。结论 [TIMP-2]×[IGFBP-7]对肾病综合征发生AKI有早期预测价值,可为临床提供诊断依据,改善患者预后。

Potential bone-inducing activity in vitro of recombinant human bone morphogenetic protein-7 from a CHO expression system

Objective: To express the recombinant human bone morphogenetic protein-7(rhBMP-7) in Chinese hamster ovary(CHO) cells, and to establish the in vitro biological activity assay of rhBMP-7.Methods: Human BMP-7 cDNA was subcloned into p114 mammalian expression vector and transfected to CHO cells by using the Lipofectamine 2000 transfection method. CHO cell supernatants were harvested and analyzed to identify the molecule mass of secreted rhBMP-7 and examine its biological activity in vitro to stimulate the synthesis of alkaline phophatase(ALP), a characteristic of osteoblast phenotypes. Results: rhBMP-7 was produced stably in CHO cells, as a processed mature disulfide-linked homodimer, with an apparent molecular mass of 36 000. Examination of the rhBMP-7 biological activity showed that rhBMP-7 specifically stimulated the production of ALP(4-fold increase at 100 ng of rhBMP-7/ml). Conclusion: The rhBMP-7 from CHO expression system has significant biological activity in induction of osteoblast phenotype, which demonstrates rhBMP-7 has the potential bone regeneration activity.

溴氰菊酯胁迫下家蚕品种辐7解毒酶基因的表达模式

昆虫体内解毒酶的过量表达和活性增强是昆虫对杀虫剂产生抗性的主要原因。辐射诱变家蚕品种辐7对菊酯类农药具有较强的耐受能力,研究溴氰菊酯胁迫下辐7体内解毒酶基因的表达模式,对于解析辐7的农药耐受性机制,培育耐农药污染的家蚕品种具有重要意义。根据不同浓度溴氰菊酯处理后辐7雄蛾的求偶和交配行为观察,最终选择2.5 mg/L溴氰菊酯处理组进行解毒酶基因的表达检测。采用荧光定量PCR检测溴氰菊酯处理后3 h和6 h辐7雄蛾触角解毒酶基因(8个羧酸酯酶基因、2个丝氨酸蛋白酶抑制剂基因、1个糜蛋白酶基因、3个脂肪酶基因、1个细胞色素P450基因和1个谷胱甘肽-S-转移酶基因)的转录水平:7个羧酸酯酶基因的表达先上调再下调,1个羧酸酯酶基因表达先下调再上调;1个细胞色素P450基因、1个丝氨酸蛋白酶抑制剂基因和1个糜蛋白酶基因表达先上调再下调;1个谷胱甘肽-S-转移酶基因、1个丝氨酸蛋白酶抑制剂基因和2个脂肪酶基因表达持续上调;1个脂肪酶基因表达在处理后6 h上调。结果表明,溴氰菊酯处理后诱导辐7雄蛾体内解毒酶基因转录水平上调,从而在一定程度上增强了辐7雄蛾对微量菊酯类农药的耐受性。

Inhibition of HOXB7 Gene Expression in Melanoma Cells by Small Interfering RNA

Objective： HOXB7 gene is a kind of transcription regulator over-expressed in malignant melanoma （MM） cell lines. It can specifically up-regulate the expression of angiogenic factors and tumor growth factors such as bFGF, GROa, VEGF and induce angiogenesis in melanoma, resulting in the proliferation and metastasis of tumor cells. We designed and synthesized HOXB7 specific siRNA to study its interfering effect on the expressions of HOXB7 and bFGF genes in melanoma A375 cell line and the biologic characteristics of A375 cells. Methods： Three synthesized siRNA with different sequences were separately transfected into A375 cells by lipofecter 2000. The expression of HOXB7 and bFGF mRNA in transfected cells was detected by RT-PCR 24 and 48 hours after transduction. The expression of bFGF protein in the transfected cells were detected by flowcytometry 48 hours after transfection. MTT assay was used to analyze the cell proliferation rate of siRNA transfected cells. Based on the in vitro experiment results, one effective siRNA sequence was selected for the construction of in vivo siRNA expression vector. Then, a malignant melanoma animal model was established. The siRNA expression plasmid was injected into the tumor foci and its influence on the growth and angiogenesis of tumor was observed. Results： The mRNA expressions of both HOXB7 and bFGF genes in the A375 cells reduced significantly 24 and 48 hour after transfection of siRNA. Expression level of the protein of angiogenic factor bFGF induced by HOXB7 gene in siRNA transfected cells was significantly lower than that in control cells 48 hours after transduction. Cell proliferation was also suppressed in siRNA transfected cells. Two of the three siRNA strands showed prominent interference effect. The in vivo study indicated that the tumor size and the microvessel density in the tumor both reduced after injection of HOXB7siRNA plasmid. Conclusion： Down-regulation of HOXB7 gene expression can effectively reduce the expression of angiogenic factor bFGF and the proliferation of MM cells. Besides, the growth and angiogenesis of MM tumor were also inhibited.

Eukaryocytic Expression of Human B7. 1 （CD80） and its Antileukemia Effect

Objective: To construct the human B7. 1 (CD80) eukaryocytic expressing vector and its expression on HL60 cells to investigate the immunotherapeutic and immunoprotective effects of B7. 1 molecule on acute leukemia. Methods: ①B7. 1 gene was subcloned from the cloning vector using PCR. Both of tlie PCR products and eukaryocytic expressing vector pHook were digested with Apa I , Sal 1 and linked using T4 DNA ligase. Tlie ligased products were transduced into DH5a. B7. 1 gene containing clones were selected by digestion with Apa I and Sal I which were further conformed by sequencing. ②HL60 cells were transformed by B7. 1 with lipofectamine and detected by FACS. ③Tumor formation, mice survival time and the immunotherapeutic and immunoprotective effects by immunization with B7. 1+ cells were evaluated. Results: ①The PCR products were about 620 bp. Six clones were all digested by Apa I and Sal I to produce 620 bp gene fragment which was in tlie same way as 67. 1. It demonstrated t/iat t/ie recombinant vector had been constructed successfully. Further sequencing confirmed the validity of the construction. There was no nucleotide mutation. ② B7. 1 was availably expressed on HL60 cells. ③ B7. 1+ HL60 cells delayed the growth of tumor and prolonged the survival time of leukemic mice, distinctively. So did tlie immunoprotection of B7. 1 + HL60 cells. Conclusion: The human B7. 1(CD80) eukaryocytic expressing vector can be successfully constructed by molecular cloned methods and can be stably and availably expressed on tlie membrane of B7. 1 negative acute myelocytic leukemia (AML) cell line HL60. Furthermore, the costimulatory molecular vaccine has significant effection of immunotherapy and immunoprotection and gives the rationale for clinical application.

The value of B7-H3 expression in expressed prostatic secretions in differential diagnosis of patients with inflammatory elevation of PSA in t-PSA gray zone

Objective To investigate the value of BT-H3 in expressed prostatic secretions ( EPS) in differential diagnosis of patients with inflammatory elevation of PSA in t - PSA gray zone ( 4 - 10 ng /ml) . Methods One hundred and sixteen patients from ages of 19 to 80 years ( mean,40 years) were studied. In the group there

IGFBP-5及p53与汉防己甲素抑制乳腺癌MCF-7细胞增殖的关系研究

目的研究汉防己甲素(tetrandrine,Tet)抑制乳腺癌MCF-7细胞增殖的作用及可能的分子机制。方法采用CCK-8、流式及Western blot分析不同浓度Tet对MCF-7细胞增殖与凋亡的影响;通过real-time PCR、Western blot检测Tet对MCF-7细胞IGFBP-5表达的影响;应用CCK-8分析Tet及IGFBP-5过表达或沉默表达对Tet抑制乳腺癌MCF-7细胞增殖作用的影响;利用Western blot检测Tet对MCF-7细胞p53表达的影响,以及过表达IGFBP-5和(或)Tet对乳腺癌MCF-7细胞p53表达的影响。结果 Tet呈浓度和时间依赖性抑制MCF-7细胞增殖,诱导G1期阻滞,并降低PCNA表达水平;同时,Tet明显增加MCF-7细胞凋亡比例,升高Bad的蛋白水平,降低Bcl-2的蛋白水平。Tet上调MCF-7细胞IGFBP-5的表达水平,过表达IGFBP-5增加Tet对MCF-7细胞的增殖抑制作用,而沉默IGFBP-5则减弱Tet的这种作用。Tet升高MCF-7细胞的p53蛋白水平,降低MDM2的蛋白水平;过表达IGFBP-5增强Tet对p53表达的促进和对MDM2表达的抑制作用。结论 Tet对MCF-7细胞增殖具有抑制作用,机制可能与其上调IGFBP-5而增强p53信号的功能有关。

LabVIEW 7 Express平台上的液压测试系统

为了满足工程实践对先进液压测试技术的需要,研究了基于美国NI公司最新推出的LabVIEW7Express虚拟仪 器开发平台的液压测试系统的软硬件结构,介绍了测试系统实现网络通讯、数据库链接和多种编程语言联合作业的方法, 提出了现代液压测试系统的技术方案。

IGFBP-7在子宫内膜腺癌中的表达及其意义

目的:研究胰岛素样生长因子结合蛋白7(IGFBP-7)在子宫内膜腺癌组织的表达,并分析其与子宫内膜样腺癌临床病理参数之间的关系。方法:选取2011年1月~2012年12月郑州大学第一附属医院病理科存档的手术切除石蜡包埋子宫内膜组织标本共123例,其中子宫内膜样腺癌61例,子宫内膜非典型增生22例,子宫内膜简单性增生10例,正常子宫内膜组织30例。免疫组化SP法检测IGFBP-7的表达,实时荧光定量RT-PCR法检测IGFBP-7 mRNA的表达。分析IGFBP-7的阳性表达率与子宫内膜样腺癌临床病理参数之间的关系。结果:(1)免疫组化法检测显示,子宫内膜样腺癌组织中的IGFBP-7阳性表达率显著高于子宫内膜非典型增生和子宫内膜简单性增生(P<0.05);IGFBP-7的表达与子宫内膜癌的病理分级、超重、合并高血压、合并糖尿病有关(P<0.05);(2)RT-PCR检测结果显示,子宫内膜样腺癌中IGFBP7 mRNA表达量显著高于子宫内膜非典型增生、简单性增生和正常子宫内膜组织(P<0.01)。结论:IGFBP-7基因可能参与了子宫内膜腺癌的发生发展过程。

绵羊IGFBP-7基因的克隆及序列分析

试验以凉山半细毛羊为研究对象,采用RT-PCR方法克隆了其IGFBP-7基因的CDS全序列,生物信息学方法深入分析其序列。结果表明,凉山半细毛羊IGFBP-7基因的CDS序列为846 bp,编码282个氨基酸,与牛、人、鼠的CDS同源性分别为99%、95%、90%,氨基酸序列同源性分别为98%、93%、89%,Gen Bank登录号为FJ589640.1;IGFBP-7基因的氨基酸分子质量为29.0 ku,理论等电点(p I)为8.25;进化分析显示其与牛、山羊等哺乳动物关系较近,与斑马鱼、鲍等亲缘关系较远;IGFBP-7基因的蛋白质疏水性区域与亲水性区域间隔较为均匀分布,有1个信号肽、2个跨膜区、16个磷酸化位点、4个N-糖基化位点和1个O-糖基化位点;二级结构分析显示无规卷曲、α-螺旋和β-折叠区域分别为64.89%、19.86%、15.25%;三级结构分析显示存在IGFBP_N和Ig-like功能域。该试验为进一步研究绵羊IGFBP-7基因的功能奠定了基础。

LabVIEW 7 Express及其应用

本文通过对虚拟仪器及面向虚拟仪器的图形化编程语言LabVIEW的简单介绍,阐述了LabVIEW7Ex-press作为最新版系统测试编程语言的特点及新特性,并以一个简单的例子说明其在编写面向虚拟仪器的应用程序方面的方法及优点。