Objective To investigate the effect of GnRH-A in gastric cavity on the function of somatostatin cells in the digestive tract of rats. Methods The immunohistochemical ABC method and enzyme-link immunoassay were used in the experiment. Results The densities of the somatostatin positive cells in the stomach and duodenum of the experiment groups were 26^6?3^893 and 51^7?2^214 respectively,and significantly lower than those in stomach 48^3?6^019 and duodenum 58^5?4^454 of control groups respectively(P

BACKGROUND: With the development and application of molecular biology and molecular genetic techniques,people research the production of human insulin by means of genetic engineering,and consider from the molecular level to reconstruct the cloning cell line which excretes insulin,so as to replace insulin injection and islet transplantation to treat diabetes mellitus.OBJECTIVE: To construct and screen human insulin gene eukaryon expression carrier of high performance.DESIGN: A randomized control experiment.SETTING: Experimental Animal Department of China Medical University.MATERIALS: Forty healthy adult Kunming mice of clean degree, weighing 20-30 g, half males and half females, were provided by the experimental animal center of Munitions University of Chinese PLA,and they were free to the access of the artificial granule food and water,and all the mice were lighted for 14 hours every day. Escherichia coli DH5α and BHK cell line were preserved by the experimental animal center.METHODS: Recombinant plasmid pEF1α-ImINS was constructed with routine method.The baby hamster kidney (BHK) cells were transfected with recombinant plasmid pEF1α-ImINS and other 3 recombinant plasmids of insulin, then screened with G418, and the positive clone cells were passaged to the 20th generation, the expressions of insulin and/or proinsulin in BHK cells were detected with radioimmunoassay and immunohistochemical technique. The PBS suspension (5×107) and the supernatant (0.5 mL) of the 20th generation BHK positive cells trasnfected with pEF1α-ImINS were injected intraperitoneally to each mice, the blood glucose was detected before and after injection to analyze the biological effect of the transgeneic product in decreasing blood glucose.MAIN OUTCOME MEASURES: The integration and expression of insulin gene in BHK cells and the changes of blood glucose before and after injection were mainly observed.RESULTS: The highest insulin expression was 7.984 mIU/L,the gray value of insulin expression was 177.50±5.10 in the cytoplasm of BHK cells, and 150.30±1.43 in the nuclear. The blood glucose at 6 hours after injection of the suspension of pEF1α-ImINS transfected BHK clone cell strain was obviously decreased,which was very significantly different from that at 24 hours before injection (P ＜ 0.01).CONCLUSION: The recombinant plasmid pEF1α-ImINS is the plasmid with the highest expression of insulin in BHK cells.The pEF1α-ImINS transfected clone cell strain has insulin expressions in the mice, and plays a role in decreasing the blood glucose of normal mice.

Objective To construct, express and identify the anti-idiotypic antibody single chain variable fragment (scFv) against Edwardsiella tarda. Methods By using RT-PCR method, the variable regions of the heavy and light chain of the anti-idiotypic monoclonal antibody (mAb) 1E11 against Edwardsiella tarda were cloned and joined with a (Gly_4ser)_3 linker, and the scFv in the orientation of V_L-linker-V_H was constructed. It was then cloned into vector plasmid pET-28a, expressed in Escherichia coli BL21(DE3), and confirmed by SDS-PAGE, Western blot and ELISA. Results The recombinant scFv could be expressed in E.coli BL21 (DE3) in a fusion protein pattern. The expression product was in the form of an inclusion body and the purified fusion protein was obtained after being purified and refolded. The SDS-PAGE and Western blot analysis showed that the molecular had the binding activity to the antigen. Conclusion The recombinant anti-idiotype scFv has been successfully constructed and expressed in E.coli BL21 (DE3), providing the basis and potential for preparation of genetically engineered vaccine against Edwardsiella tarda.

Objective To construct,express and identify the anti-idiotypic antibody single chain variable fragment(scFv) against Edwardsiella tarda.Methods By using RT-PCR method,the variable regions of the heavy and light chain of the anti-idiotypic monoclonal antibody(mAb) 1E11 against Edwardsiella tarda were cloned and joined with a(Gly4Ser)3 linker,and the scFv in the orientation of VL-linker-VH was constructed.It was then cloned into vector plasmid pET-28a,expressed in Escherichia coli BL21(DE3),and confirmed by SDS-PAGE,Western blot and ELISA.Results The recombinant scFv could be expressed in E.coli BL21(DE3) in a fusion protein pattern.The expression product was in the form of an inclusion body and the purified fusion protein was obtained after being purified and refolded.The SDS-PAGE and Western blot analysis showed that the molecular weight of scFv protein was 27 ku.Indirect ELISA confirmed that the scFv had the binding activity to the antigen.Conclusion The recombinant anti-idiotype scFv has been successfully constructed and expressed in E.coli BL21(DE3),providing the basis and potential for preparation of genetically engineered vaccine against Edwardsiella tarda.

This paper is to report our experience of using the internal fixation with compression-screw to treat 9 fractures of carpal navicular bone including two fresh) two old and 5 non- union cases.The mean follow-up time of the 9 cases was one year. Two fresh and 2 old fractures healed uneventfully and so did the 2 cases out of the 5 non-unions. The symptoms, however, disappeared in all the cases and the function of the wrist was or nearly was normal. All the patients resumed their original work.It is believed that this method of internal fixation with compression-screw is simple and easy to perform. Its effect is reliable. No external fixation is needed after the operation. And the function of the wrist can be restored after a short period of treatment.It is suggested that the indications of this operative treatment are non-union cases without ischemic necrosis, unstable fresh fractures with displacement and fractures in the proximal third or half of the navicular bone.

Objective To study the value of intensive care nursing scoring system(ICNSS) on care resource allocation in intensive care unit(ICU). Methods 108 ICU patients were divided into the observation group (55 cases) and the control group (53 cases) randomly. In the observation group,the ICNSS scale was applied to evaluate nursing workload, and care resource was allocated according to intensive care nursing scoring. In the control group, care resource was allocated according to dynamic monitoring of acute physiology and chronic health evaluation (APACHE Ⅱ ) scoring. The ICU monitoring time,medical cost,incidence of complications during the hospitalization,satisfaction of the nurses and the patients or their relatives were compared between two groups. Results The ICU monitoring time,medical cost and incidenceof complications during the hospitalization in the observation group were significantly less than those in the control group, while the satisfaction of the nurses and the patients or their relatives in the observation group was markedly better than that in the control group. Conclusions Care resource allocation according to ICNSS is worthy of promotion and application in ICU because it can more significantly improve nursing quality,satisfaction of nurses and patients or their relatives than care resource allocation according to APACHE Ⅱ scoring.

Cell smears were made from fresh mammary tissue during operation. There are 32 samples with definite pathological diagnosis. DNA content was measured in interphase nuclei of single epithelial cells of lesions of the human breast by a microspectrophotometric technique. The normal lymphocytes from 2 cases of normal blood samples and from 4 cases of mammalopathic tissue were used as a reference for the DNA value corresponding to a diploid number of chromosomes. The normal lymphocyte had mean DNA value in 11.2?0.2 A.U. and 10.9?1.3 A.U. The rest of the measurements showed: 12.1?1.3 A.U. in the, epithelial nuclei of 10 cases of neighbouring mammalopathic tissue; 22.6?1.0 A.U. in 9 cases of fibrcma of the breast; 31.9?2.3 A.U. in 8 cases of carcinomas of the breast. The cell nuclei of carcinomas showed abnormal DNA values and the multiploid cells were dominant. It's histograms showed a shift to right and either a double-peak or multiploid-peak distribution. The DNA contents of the benign lesion cells were significantly lower than that of cancer cells, but higher than that of normal cells. The peak of their histogram showed they are between 2C-4C.

Objedctive To study the mechanism of GnRH agalogus(Alarelin)on[ Ca2 + ]; mobilization in stomach smooth muscle cells (SSMc)of rats. Methods Cells were cutured and loaded with Fluo-3-AM. [Ca2 + ]; was measured by nuomeent intensity(FL) in each cell with confocal micoopy. Results (1 ) In Hanks solution. l0-7, l0-6, l0 - 5mol L- 1 Alarelin can elevate[ Ca2 + ], its peak - resting values reached 6. 00 ?0. 50 .9. 23 ?0. 62. l8. 97 ?2. 42 respectively, which indicate that the level of [Ca2+ ]; act in an deede- pendent and time - dependent. (2) thntredly. when Alarelin was 10 4mol?Ｌ－１, its peak - resting value only reached 6. 32 ?0 .67, conpared with Alarelin l0 5m.l L－1 which was sigificantly lower(P 0. 05 ). (5 )When pertreatment with Lacidipine in Hanks solution, the effect of Alarelin l0 5mol L-1.as partly inhibited(P

Objective To investigate the expression of TNF-related apoptosis-inducing ligand(TRAIL) and TNF-related apoptosis-inducing ligand receptor(TRAILR) in thyroid carcinoma. Methods Expression and distribution of TRAIL and TRAILR in thyroid tissues from 13 patients with papillary thyroid carcinoma, 3 patients with follicular thyroid carcinoma and 5 normal necrospy subjects were studied by immunohistochemical methods. Results TRAIL and all of TRAILR immunoreactivity was observed in both thyroid tissues from papillary thyroid carcinoma and follicular thyroid carcinoma. TRAILR4 was expressed highly in all thyroid carcinoma tissues but weakly in normal thyroid tissues.Conclusion TRAIL and all of TRAILR were present in thyroid tissues of papillary thyroid carcinoma and follicular thyroid carcinoma. Expression of TRAIL, TRAILR1 and TRAILR2 by thyrocytes in thyroid carcinoma may induce their apoptosis through autocrine or paracrine. Thyroid carcinoma cells were sensitive to TRAIL-inducing apoptosis despite expression of both decoy receptors. [

Objective To investigate the effect of GnRH-A in gastric cavity on function of gastrin clls in digestive tract of rats. Methods The immunohistochemical ABC method and enzyme-link immunoassay were used in the experiment. Results The densities of the gastrin immunoreactivity cells in the stomach and small instestin of the experiment groups were 19.60?3.63 and 18.00?2.31 respectively, those in stomach was 10.30?2.41 while in small intestin was 9.00?2.05 of control groups respectively (P