The distribution and morphology of 5-HT immunoreactive endocrine cells in the gastrointestinal tract of 5 adult rats were studied by the immunohistochemicael PAP method with nickel-intensified DAB on paraffin sections of intestine rolls. The density of 5-HT immunoreactive endocrine cells in the gastrointestinal tract of the rat is highest in the pylorus, duodenum and colon and moderate in the jejunum, ileum, caecum and rectum and lowest in the body of the stomach. The 5-HT immunoreactive endocrine cells are various in shape. Some of them have several processes extending between other epithelial cells. The basal portion of some endocrine cells have processes with 5-HT positive substance accumulating in their ends. The processes of the basal portion of some endocrine cells extend into lamina propria through the basal membrane. The 5-HT positive substance of many endocrine cells can be found to extend to luminal surface of the crypt or intestinal tract. These results indicate that the 5-HT immunoreactive endocrine cells can release 5-HT by both endocrine and exocrine ways.

Localization and quantitation of GnRH, somatostatin (SS), and ?-endorphin (?-EP) in human placentalvilla were studied using immunogold-silver staining method. GnRH and SS immunoreactive positive substance existed in the cytotrophoblasts of many placental villi and in syncytiotrophoblasts of few placental villi, ?-EP immunoreactive positive substance localized in syncytiotrophoblasts of many placental villi and in cytotrophoblasts of few placental villi. These results suggest that the cytotrophoblasts and syncytiotrophoblasts are able to synthesize these three kinds of regulatory peptides, however, SS and GnRH may be synthesized mainly in the cytotrophoblasts, ?-EP mainly be synthesized in the syncytiotrophoblasts. The amount of GnRH and ?-EP in placenta villi show obvious change with progress of pregnancy. This change show a negative relationship between the GnRH and ?-EP, which suggested that there may be a functional reciprocal inhibition between the GnRH and ?-EP.

Objective Uncemented prothesis has become the preference of first hip arthroplasty, but there is a little study on its application in cemented acetabular revision arthroplasty.This paper aimed to evaluate the clinical effects of uncemented acetabular cup in cemented acetabular prosthesis revision arthroplasty. Methods A retrospective analysis was made on the clinical data of 31 patients(31 hips) who underwent revision arthroplasty using uncemented acetabular prosthesis from January 2012 to March 2015.Har-ris hip score( HSS) was applied to measure the hip function and visual analogue score( VAS) was preformed to assess the pain degree of knee joint preoperatively and postoperatively(3 months, 6 months and 1 year).All the patients were followed up for a mean of 22 months ranging from 3 to 42 months. Result The mean HSS increased from preoperative points (53.19 ±9.12) to postoperative points([77.71 ±5.75]at 3 months,[80.61 ±5.74] at 6 months,[82.94 ±5.80] at 1 year ).The mean VAS score decreased from preoperativepoints (6.23 ±1.23) to postoperative points(0.26 ±0.27).No lossening, infection or deep vein thrombosis were found in the patients′prostheses. Conclusion For patients with PaproskyI-Ⅱ acetabular defects, uncemented acetabular prosthesis has satisfactory short-term clinical results in revision arthroplasty with cemented acetabular prosthesis, however, long-term clinical results remain to be further observed.

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 study whether gonadotropin releasing hormone receptor (GnRHR)mRNA exist in rat submaxillary gland and it's gene sequence. Methods\ The total RNA isolated from rat submaxillary gland was amplified by RT\|PCR, the PCR products were identified by sequencing with Sanger's method. Results\ The specific amplified band of GnRHR mRNA was detected through agarose gel electrophoresis and gene sequence is identical to the sequence of GnRHR which has been reported in rat pituitary. Conclusion\ GnRHR can be produced by submaxillary and response for modulating biological function of submaxillary.\;

Objective To investigate the mutations of basal core promoter (BCP) and precore (PreC) region of hepatitis B virus (HBV) and the association with the development of hepatocellular carcinoma in patients with chronic HBV infection.Methods Totally 381 untreated HBV patients were recruited from the Department of Infectious Diseases,People's Hospital of Shangyu from Jan 2003 to Dec 2010,which included patients with chronic hepatitis B (CHB,n =166),cirrhotic hepatocellular carcinoma (cirrhotic-HCC,n =158) and noncirrhotic hepatocellular carcinoma (noncirrhotic-HCC,n=57).The mutations in HBV BCP and PreC and the genotypes of HBV were determined by polymerase chain reaction (PCR) and direct sequencing.Data were analyzed by chi square test and Logistic regression.Results The HBV genotype of most cases was genotype B (CHB,n =124;cirrhotic-HCC,n=126 ; noncirrhotic-HCC,n=50).In univariant analysis,BCP V1753 (x2 =7.927,P=0.005),BCP T1762/A1764 (x2 =12.796,P＜0.01),PreC A1896 (x2 =6.890,P=0.009) and PreC A1899 (x2=11.850,P =0.001) mutations were more frequently detected in cirrhotic-HCC patients than those in CHB patients.PreC A1896 (x2 =27.310,P＜0.01) and A1899 (x2=7.575,P=0.006) mutations were highly detected in noncirrhotic-HCC patients than those in CHB patients.Multivariate Logistic regression analysis revealed that in HBeAg positive patients,BCP T1762/A1764 (wald=6.180,P=0.016,OR=8.883) and PreC A1899 (wald=10.279,P=0.001,OR=7.475) mutations were independently associated with the development of cirrhotic-HCC; PreC A1896 (wald=4.324,P=0.038,OR=4.439) and PreC A1899 (wald=4.850,P=0.028,OR=6.010)mutations were independently associated with the development of noncirrhotic-HCC.While in HBeAg negative patients,PreC A1896 mutation (wald=15.448,P＜0.01,OR=12.128) was independently associated with the development of noncirrhotic-HCC.Conclusions BCP T1762/A1764 mutations are associated with the development of cirrhotic-HCC in HBeAg positive patients.PreC A1896 mutation is associated with the development of noncirrhotic-HCC in HBeAg positive and HBeAg negative patients.PreC A1899 mutation is associated with the development of cirrhotic-HCC and noncirrhotic-HCC in HBeAg positive patients.

Objective To detect the existence of gonadotropin-releasing hormone receptors(GnRH-R) in rat liver,and to supply morphological evidence for studying the functional significance of the GnRH in hepatocyte. Methods Immunohistochemical SABC method and in situ hybridization with a digoxigenin labeled probe were used to study the localization of GnRH-R in the livers of five rats. Results The rat hepatocytes were found with the GnRH-R immunoreactivity and GnRHR mRNA hybridization signals.The GnRH-R immunoreactive substance was distributed in the cytoplasm of all positive cells,with immuno-negative nuclei.The GnRHR mRNA hybridization signals were also detected in the cytoplasm of all positive cells but not in the nuclei.Hepatocytes in different parts of liver lobules showed different intensity of GnRH-R immunoreactivity and GnRH-R mRNA hybridization signals.Conclusion The rat hepatocyte may express GnRH-R.The hepatocytes in different parts in liver lobules exhibit different levels of GnRH-R expression.

Objective To investigate the distribution of folliclestimulating hormone (FSH),luteinizing hormone (LH) and colocalization with gonadotropin releasing hormone receptor (GnRHR) in rat submaxilary glands. Methods Distribution of FSH, LH and colocalization with GnRHR consecutive sections of rat submaxilary glands were investigated by immunohistochemical colocalization methods. Results FSH and LH immunoreactivity were observed in the epithelial cells of serous acinus, secretory tubes, excretory ducts and granular convoluted tubule. The immunoreactive materials were brown and distributed in the cytoplasma with negative nucleolus. The results of immunohistochemical colocalization showed not only FSH but also GnRHR immunoreactivity in the same structure of two adjacent section. The distribution of the positive substance of FSH and GnRHR were similar to each other. The most of showing GnRHR immunoreactivity cells were detected LH immunoreactivity in the same structure of two adjacene section and the others were immunonegative. The GnRNR immunoreactive materials were distributed in the cytoplasma with negative nucleolus.Conclusion The epithelial cell of serous acinus, secretory tubes, excretory ducts and granular convoluted tubule of rat submaxilary glands may be synthesized and secreted FSH and LH. These cells with FSH and LH positive immunoreaction of rat submaxilary glands may be regulated by Gonadotropin releasing hormone (GnRH) through autocrine or paracrine.;

Objective In this study, we investigated the distribution of Peroxiredoxin II mRNA in follicles at all stages in mouse ovaries and mouse secondary oocytes(MII eggs) basing on our previous researches, to provide the morphological basis for exploring the effect of Peroxiredoxin II on oogenesis and oocyte maturation in the mouse. Methods To observe the distribution of Peroxiredoxin II mRNA in follicles at all stages in ovaries and the localization of Peroxiredoxin II mRNA in Germinal-Vesicle intact oocytes(GV oocytes) and MII eggs by in situ Hybridization. Results In situ hybridization of ovary section revealed that the signals for Peroxiredoxin II mRNA were undetectable in oocytes of primordial follicles, and moderate signals for Peroxiredoxin II mRNA were observed in oocytes of primary follicles. Moreover, strong signals for Peroxiredoxin II mRNA were evident in antral follicles. The signals for Peroxiredoxin II mRNA also existed in GV oocytes and MII eggs in vitro. The hybrid signals were stronger in GV oocytes than in MII eggs. In addition, the weak but consistent signals for Peroxiredoxin II mRNA were detected in follicular cells from primordial follicles to large antral follicles. Peroxiredoxin II mRNA was located in cytoplasm of oocytes and follicular cells, but not in nuclei.Conclusion These results suggested that Peroxiredoxin II might be involved in the regulation of oogenesis and oocyte matruation in the mouse.;