Central role of GRP78 in growth of gastric carcinoma cells / 中国病理生理杂志

AIM:To explore the effect of glucose-regulated protein 78 (GRP78) on the gastric carcinogenesis. METHODS:GPR78 expression patterns were examined in 34 specimens from gastric carcinoma patients using the immu-nohistochemistry (IHC) assay, and in 10 specimens using Western blotting analysis .In addition, the expression of GPR78 and cyclin D1 was detected in human gastric cancer cell lines SGC 7901 and SGC7901-H78 (overexpressing GRP78) by Western blotting.RESULTS:By IHC assay, GRP78 was found to be highly expressed in the cytoplasm of gastric carcino-mas as compared with the adjacent non-malignant tissues and corresponding normal tissues .GRP78 expression was positive-ly correlated with gender and histological differentiation (P0.05).Furthermore, we found that with the increased expression of GRP 78 in SGC7901-H78 cells, the expres-sion of cyclin D1 was also elevated .CONCLUSION:GRP78 might be a key player to be involved in the growth of gastric cancer.

Construction of lentiviral vector-mediated siRNA knockdown of ER-α36 and its action on gastric cancer cell growth / 中国病理生理杂志

[ ABSTRACT] AIM:To construct a lentiviral vector for stable delivery of the ER-α36 gene and to detect its effect on SGC7901 cell growth.METHODS: The efficient RNAi targeting sequences identified for the ER-α36 gene were screened.The Oligo DNA was synthesized with target sequences and annealed to form double-stranded DNA.Then it was digested by Xho I and EcoR I and connected with GV307 vector to produce LV-ER-α36-RNAi lentiviral vector.PCR was used to screen the positive clones and sequence.The LV-ER-α36-RNAi, pHelper 1.0 and pHelper 2.0 plasmids were co-transfected into 293T cells for producing lentiviral vector and infecting SGC7901 cell line.Fluorescence microscopy, real-time PCR and Western blotting were used to detect the transfection efficiency and gene silencing effect.17β-estrodial at concentration of 1 ×10 -10 mol/L was used to stimulate the recombinant cell line, and the action on the growth of gastric cancer cells and the expression of Src, ERK1/2 and cyclin D1 were determined.RESULTS: DNA sequencing analysis confirmed the identity of recombinant shRNA expression vectors.Immunofluorescence assay demonstrated that transfection efficiency was above 80%.Transfection of LV-ER-α36-RNAi significantly knocked down the expression of ER-α36 at mR-NA and protein levels with tetracycline ( TeT) simulating as revealed by real-time PCR and Western blotting.Compared with control group, the growth of the recombinant cell line declined and the expression of Src, ERK1/2 and cyclin D1 and the activation of Src decreased (P<0.05).CONCLUSION: Lentiviral vectors that silence ER-α36 expression are con-structed successfully and can be used to study the role of ER-α36 in gastric cancer.The ER-α36 is related with many kinds of cancer cell growth, including gastric cancer cells.

Construction of mixed ability in eight-year program clinical teaching of obstetrics and gynecology / 中华医学教育探索杂志

Huazhong University of Science and Technology is one of the five schools that trial implementing eight-year program medical education of first batch.Eight-year program medical students in Tongji college came into obstetrics and gynecology clinical contact in 2010.According to the characteristics of students and discipline,obstetrics and gynecology department used multiple teaching methods and means as PBL teaching,integrated case management,typical clinical case conference,clinical diagnosis and treatment,academic lecture,research report,experimental design,scientific research and experi-ment,etc.Available practice mode was reformed.Those students accomplished clinical practice,and achieved expected effect.

Determination of the Early Time of Death by Computerized Image Analysis of DNA Degradation: Which Is the Best Quantitative Indicator of DNA Degradation? / 华中科技大学学报（医学）（英德文版）

This study evaluated the correlation between DNA degradation of the splenic lymphocytes and the early time of death, examined the early time of death by computerized image analysis technique (CIAT) and identified the best parameter that quantitatively reflects the DNA degradation.The spleen tissues from 34 SD rats were collected, subjected to cell smearing every 2 h within the first 36 h after death, stained by Feulgen-Van's staining, three indices reflecting DNA content in splenic lymphocytes, including integral optical density (IOD), average optical density (AOD), average gray scale (AG) were measured by the image analysis. Our results showed that IOD and AOD decreased and AG increased over time within the first 36 h. A stepwise linear regression analysis showed that only AG was fitted. A correlation between the postmortem interval (PMI) and AG was identified and the corresponding regression equation was obtained. Our study suggests that CIAT is a useful and promising tool for the estimation of early PMI with good objectivity and reproducibility,and AG is a more effective and better quantitative indicator for the estimation of PMI within the first 36 h after death in rats.

The in vitro antitumor effect and in vivo tumor-specificity distribution of human-mouse chimeric antibody against transferrin receptor.

Transferrin receptor (TfR/CD71) deserves attention as a selective target for cancer therapy due to its higher expression in tumors versus normal tissues. Also, it has been shown the mouse-derived monoclonal antibody against TfR can significantly inhibit the proliferation of tumor cells. Through constructing the chimeric antibody against TfR, the antigenicity of antibody can be weakened, and most importantly, the antitumor effect of antibody can be strengthened by the introduction of the human Fc fragment. In previous studies, we successfully constructed the human-mouse chimeric antibody against TfR (D2C) and demonstrated that its Fab fragment could specially recognize the TfR on the surface of target cells. In this study, through labeling the chimeric antibody D2C with 125I, we calculated the affinity constant (Ka) of 9.34-9.62x10(9) l/mol for this antibody according to the Scatchard drawing method. Moreover, in vivo studies in nude mice-bearing human liver cancer (SMMC-7721) xenografts have shown that the radioactivity distribution ratio of 131I-D2C on T/NT was 2-14:1 or 3-21:1 on the seventh day after intraperitoneal or intratumoral injection of 131I-labeled D2C (131I-D2C). These evidences indicated that the in vivo distribution of D2C display the characteristics of certain tumor-specificity localization. In vitro studies, D2C can induce the apoptosis of K562 through the mitochondria death pathway and arrest the cell at G1 phase, as determined by cell cycle analysis. Using the human tumor cells (K562, CEM, and SMMC-7721) expressing TfR as target cells, and normal human PBMC as effector cells, Fc fragment of D2C can perform both the antibody-dependent cell-mediated cytotoxicity and the complement-dependent cytotoxicity. Together, it was demonstrated that the D2C display a tumor-specificity distribution, and has a strong antitumor effect. Thus, it has the potential therapeutic significance.

Correlative Analysis on the Relationship between PMI and DNA Degradation of Cell Nucleus in Human Different Tissues / 华中科技大学学报（医学）（英德文版）

To determining the postmortem interval (PMI) through quantitative analysis of the DNA degradation of cell nucleus in human brain and spleen by using image analysis technique (IAT). The brain and spleen tissues from 32 cadavers with known PMI were collected, subjected to cell smear every 1 h within the first 5-36 h after death, stained by Feulgen-Van's staining, Three indices reflecting DNA in brain cells (astrocytes) and splenic lymphocytes, including integral optical density (IOD), average optical density (AOD), average gray (AG) were measured by employing the mage analysis instrument. The results showed that IOD and AOD declined and AG increased with the prolongation of dead time within 5-36 h. A correlation between the PMI and gray parameters (IOD,AOD and AG) was identified and the corresponding regression equation was obtained. The parameters (IOD,AOD and AG) were proved to be effective quantitative indicators for accurate estimation of PMI within 5-36 h after death.

Correlative analysis on the relationship between PMI and DNA degradation of cell nucleus in human different tissues / 华中科技大学学报（医学）（英德文版）

To determining the postmortem interval (PMI) through quantitative analysis of the DNA degradation of cell nucleus in human brain and spleen by using image analysis technique (IAT). The brain and spleen tissues from 32 cadavers with known PMI were collected, subjected to cell smear every 1 h within the first 5-36 h after death, stained by Feulgen-Van's staining, Three indices reflecting DNA in brain cells (astrocytes) and splenic lymphocytes, including integral optical density (IOD), average optical density (AOD), average gray (AG) were measured by employing the mage analysis instrument. The results showed that IOD and AOD declined and AG increased with the prolongation of dead time within 5-36 h. A correlation between the PMI and gray parameters (IOD, AOD and AG) was identified and the corresponding regression equation was obtained. The parameters (IOD, AOD and AG) were proved to be effective quantitative indicators for accurate estimation of PMI within 5-36 h after death.

Prognostic significance of the number of lymph nodes removed surgically in patients with colorectal cancer / 中国普通外科杂志

or ≤50 in each patient. 5-year survival curves were estimated with the Kaplan-Meier method and compared by the log-rank test. ResultsWT5”BZ Analysis revealed that among node-negative patients, the 5-yerar's survival rate was 23% higher in patients with more than 50 nodes removed than those with less than 50. For node-positive patients, the difference of 5-year′s survival rate between the related two subgroups reached 36%.WT5”HZConclusion The number of lymph node resected has great impact on the long term survival of patients with colorectal cancer.

The relationship between the resection extent and number of lymph node dissection and the effect of metastatic lymph node number on the prognosis of proximal gastric cancer / 中国普通外科杂志

Objective To study the relation between the surgical extent and number of lymph node dissection and the effect of metastatic lymph node number on the prognosis of proximal gastric cancer. Method Thirty-one patients with proximal gastric cancer underwent D_2 or D_3 operation, and lymph node were harvested from specimen. The number of lymph node dissection in proximal gastrectomy was compared with that in total gastrectomy. Results A total of 1971 lymph nodes were obtained from the 31 specimens, the mean was 63 per case. There were 57 lymph nodes per case for patients with proximal gastrectomy and 71 per case with total gastrectomy,64 per case with left half pancreaticosplenectomy and 63 per case with splenectomy. According to the new 5th N stage system, the 5 year survival of N_1,N_2,N_3 were 36%, 11%, and 0 respectively. Conclusion Along with the wide invasion of the carcinoma wide resection with extended lymph node dissection is mandatory, reserved left half of the pancreas has no influence on the number of harvested lymphnode, the new quantitative N staging is superior to the old in predicting the prognosis.

Study on relationship between changes of DNA content in human spleen nuclei and the postmortem interval by image analysis / 中国法医学杂志

Objective To study changes of DNA content in human spleen nuclei and seek an experimental method for estimation of the postmortem interval(PMI)using computerized image-analysis technique(CIAT).Methods Smear sections from spleen sampled were collected in 36 cadavers with known the accurate PMI respectively every hour within the first 36 hours after death,which were then fixed with cold Carony fixation.The smeared sections were stained by Feulgen-van's staining method.3 indices for spleen nucleic DNA including integral optical density(IOD),average optical density(AOD)and average gray(AG)were measured using the CIAT.Results IOD and AOD in the spleen nuclei declined regularly,whereas AG increased with the extension of PMIs in 36 hours.Conclusion There are definite relationships between the PMI and gray parameters(IOD、AOD and AG)representing the DAN content of nucleic DNA in the spleen in 36 hours after death,which may be used for estimation of PMI.