人肝细胞癌中转移抑制基因nm23编码蛋白的表达

目的探讨肝脏冷保存与移植肝原发性无功能的关系。方法采用具有免疫耐受性的同种大鼠原位肝移植模型进行研究。将鼠肝分别在４℃ＵＷ液中保存１，６和２４ｈ，分别在移植后１ｈ和２４ｈ取肝脏做病理观察。结果保存在ＵＷ液中的肝脏基本保存了正常的组织结构，而在移植后则不同程度地出现了变性或坏死，并且肝细胞的坏死是以小叶中央静脉为中心。保存时间越长，病变越严重。结论肝脏冷保存时间与移植肝原发性无功能的发生密切相关。     

Copper transportion of WD protein in hepatocytes from Wilson disease patients in vitro

AIM:To study the effect of copper transporting P-typeATPase in copper metabolism of hepstocyte andpathogenesis of Wilson disease(WD).METHODS:WD copper transporting properties in someorganelles of the cultured hepstocytes were studied fromWD patients and normal controls.These culturedhepstocytes were incubated in the media of copper 15mg·L~(-1) only,copper 15 mg·L~(-1) with vincristine(agonist of P-type ATPase)0.5mg·L~(-1),or copper 15 mg·L~(-1) withvanadate(antagonist of P-type ATPase)18 39mg.L~(-1)separately.Mlcrosome(endoplasmic reticulum and Golgiapparatus),lysosome,mitochondria,and cytosol wereisolated by differential centrifugstion.Copper contents inthese organelles were measured with atomic absorptionspectrophotometer,and the influence in copper transportionof these organelles by vanadate and vincristine werecomparatively analyzed between WD patients and controls.WD copper transporting P-type ATPase was detected bySDS-PAGE in conjunction with Western blot in liver samplesof WD patients and controls.RESULTS:The specific WD proteins(Mr155 000 lanes)wereexpressed in human hepatocytes,including the control andWD patients.After incubation with medium containingcopper for 2 h or 24 h,the microsome copper concentrationIn WD patients was obviously lower than that of controls,and the addtion of vanadste or vincristine would change thecopper transporting of microsomes obviously.Whenincubated with vincristine,levels of copper in microsomewere significantly increased,while incubated with vanadate,the copper corcentrations in microsome were obviouslydecreased.The results indicated that there were WDproteins,the copper tmnsportion P-type ATPase in themicrosome of hepstocytes.WD patients possessedabnormal copper transporting function of WD protein in themicrosome,and the agonist might correct the defect of copper transportion by promoting the activity of coppertransportion P-type ATPase.CONCLUSION:Copper tmnsportion P-type ATPase plays animportant role in hepatocytic copper metabolism.Dysfunction of hepatocytic WD protein copper transportionmight be one of the most important factors for WD.     

胃癌MG-7抗原模拟表位与HSP70融合基因的构建及表达

目的构建和表达胃癌MG7模拟表位与人热休克蛋白70(HSP70)的融合基因.方法采用加端PCR的方法,将MG7模拟表位(GC23)的编码序列融合到HSP基因的3′端;PCR获得的融合基因片段经TA克隆法克隆到pUCm-T载体上,并进行DNA测序;将融合基因片段从pUCm-T载体上酶切后,亚克隆到表达载体PET-21a(+)上;将重组质粒PET-21a(+)/GC23-hsp转化大肠杆菌,经IPTG诱导后,收集细菌,菌体裂解后进行SDS-PAGE及Western blot检测结果应用PCR方法扩增出约2.0kb的目的片段,序列测定结果证实,模拟表位GC23序列成功地融合到HSP70基因的3′端,GC23及HSP70基因序列均正确;经EcoRⅠ+XhoⅠ酶切及PCR鉴定证实,融合基因成功地克隆到原核表达载体PET-21a(+)上;转入重组质粒的大肠杆菌经IPTG诱导后,进行SSS-PAGE发现在Mr为72000处有表达量明显增多的蛋白条带,Western blot证实,Mr72000处的条带为目的条带.结论成功构建并表达了MG7模拟表位与HSP70的融合基因.     

胃癌D17S261和D17S799位点二核苷酸重复序列不稳定性的意义

目的研究二核苷酸重复序列不稳定性〔ＤＲＳＩ〕在胃癌发生中的作用及其临床意义．方法采用ＰＣＲ方法检测了Ｄ１７Ｓ２６１和Ｄ１７Ｓ７９９位点二核苷酸重复序列不稳定性．结果胃癌总ＤＲＳＩ发生率为３４％（１７／５０），其中高中分化腺癌ＤＲＳＩ阳性率（６６７％，１０／１５）显著高于低分化癌（１９４％，６／３１，Ｐ＜００１）；肠型胃癌ＤＲＳＩ阳性率（５５６％，１０／１８）显著高于胃型胃癌（２０％，６／３０，Ｐ＜００５），ＤＲＳＩ与胃癌部位、大小、浸润、分期、淋巴结转移无显著相关．结论ＤＲＳＩ在胃癌的发生中可能起重要作用．     

Identification of differentially expressed genes in normal mucosa, adenoma and adenocarcinoma of colon by SSH

Abbreviation CRC colorectal cancer.SSHsuppression subtrsctive hybridization.LD PCR longdistance polymerase chain reaction.HLA human leukocyteantigen.IGRBP insulin-like growth factor binding protein.GN guanylin,EF1 elongation factor 1AIM To construct subtracted cDNA libraries and furtheridentify differentially expressed genes that are related tothe development of colorectal carcinoma(CRC).METHODS Suppression subtractive hybridization(SSH)was done on cDNAs of normal mucosa,adenoma andadenocarcinoma tissues from the same patient.Threesubtracted cDNA libraries were constructed and thenhybridized with forward and backward subtracted probesfor differential screening.Positive clones from eachsubtracted cDNA library were selected for sequencing andBLAST analysis.Finally,virtual Northern Blot confirmedsuch differential expression.RESULTS By this way,there were about 3-4×10~2clones identified in each subtracted cDNA library,inwhich about 85% positive clones were differentiallyscreened.Sequencing and BLAST homology searchrevealed some clones containing sequences of knowngene fragments and several possibly novel genes showingfew or no sequence homologies with any knownsequences in the database.CONCLUSION All results confirmed the effectiveness andsensitivity of SSH.The differentially expressed genesduring the development of CRC can be used to shed lighton the pathogenesis of CRC and be useful genetic markersfor early diagnosis and therapy.     

食管癌患者O6-烷基鸟嘌呤-DNA烷基转移酶基因多态性研究

目的　检测中国食管癌高发区居民和高加索居民AGT     

Expression of liver cancer associated gene HCCA3

AIM:To study and clone a novel liver cancer related gene,and to explore the molecular basis of liver cancer genesis.METHODS:Using mRNA differential display polymerssechain reaction(DDPCR),we investigated the difference ofmRNA in human hepstocallular carcinoma(HCC)and pairedsurrounding liver tissues,and got a gene probe.Byscreening a human placenta cDNA library and genornichomologous extend,we obtained a full-length cDNA namedHCCA3.We analyzed the expression of this novel gene in 42pairs of HCC and the surrounding liver tissues,anddistribution in human normal tissues by mssns of Northernblot assay.RESULTS:A full-length cDNA of liver cancer associated geneHCCA3 has been submitted to the GeneBank nucleotidesequence databases(Accession No.AF276707).Thepositive expression rate of this gene was 78.6%(33/42)inHCC tissues,and the clinical pathological data showed thatthe HCCA3 was closely associated with the invasion oftumor capsule(P=0.023)and adjacant small metastasissatellite nodules lesions(P=0.041).The HCCA3 was widelydistributed in the human normal tissues,which wasIntensively expressed In lungs,brain and colon tissues,while lowly expressed In the liver tissues.CONCLUSION:A novel full-length cDNA was cloned anddifferentiated,which was highly expressed in liver cancertissues.The high expression was closely related to thetumor invssivensss and metastasis,that may be the lateheredited change in HCC genssis.     

小儿消化性溃疡遗传性及遗传度的研究

目的研究遗传因素在小儿消化性溃疡（ＰＵ）发病中的作用．方法内镜诊断３～１４岁ＰＵ患儿５２例，抽血测定血型，并随访患儿的遗传度及家族发病情况．同时问卷调查了本市３所小学１１１９位学生家系中一级亲属ＰＵ发病率，按照遗传度计算公式求出小儿ＰＵ的遗传度．结果儿童ＰＵ中多见的是十二指肠溃疡，首次发病年龄＜９岁占４８％，男性患者约为女性２倍，体形多为瘦长形，体块指数＜４５占６７３０％，Ｒｈ＋Ｏ血型人占４６１５％，有ＰＵ家族史为４８０８％，患者一级亲属ＰＵ发病率约为群体的１１倍，其遗传度为１０８６６％．结论小儿ＰＵ与成人不完全相同，遗传因素在本病的发生中起着十分重要的作用．     

Expression of the c-erbB-2 proto-oncogene product in gastric carcinoma and precancerous lesions

AIM: To study the relationship between the expression of the c-erbB-2 proto-oncogene product with gastric mucosal carcinogenesis and the behavior of gastric carcinoma.METHODS: Specimens from nine normal gastric mucosa, 23 gastric mucosal dysplasia (10 slight, six moderate, seven severe), 18 early gastric carcinoma, and 30 advanced gastric carcinoma were marked with P₁₈₅ monoclonal antibody using the immunohistochemical peroxidase-avidin-biotin complex method. The relation between P₁₈₅ expression with histological type, size, and lymph node metastasis of gastric carcinoma were analyzed.RESULTS: Normal gastric mucosa was negative for P₁₈₅; Only a few cells in the neck region of the mucosal glands were very weakly positive. Relatively high positive rates were found in the slight, moderate, and severe dysplasia specimens (50%, 83.3%, and 85.7%, respectively). A 22.2% and 56.7% P₁₈₅-positive rate was found in early gastric carcinoma and in advanced gastric carcinoma, respectively. Statistically, the P₁₈₅-positive rates in severe dysplasia and advanced gastric carcinoma were significantly higher than that in early gastric carcinoma (P < 0.05). The P₁₈₅-positive rate in the group with lymph node metastasis was significantly higher than that of the group without lymph node metastasis (59.3% vs 23.8%, P < 0.05), but P₁₈₅ expression was not related to histological type and size of gastric carcinoma.CONCLUSION: The c-erbB-2 proto-oncogene might participate in gastric mucosal proliferation, repair, and carcinogenesis, and gastric carcinoma with P₁₈₅ expression might have a stronger potential of infiltration and metastasis.