Objective To summarize our experience in the management of stones impacted at cystic duct during laparoscopic cholecystectomy(LC). Methods There were 58 patients with stones impacted at cystic duct during LC from July 1997 to June 2001.Cystic duct was incised to remove stones first,then intraoperative cholangiography was performed.If stones were found in common bile duct,they would be taken out by intraoperative endoscopic sphincterotomy or open operation. Results All the impacted stones in 58 patients were removed successfully.51 coses underwent LC.Intraoperative cholangiography showed there were common bile duct stones in 7 cases.For these 7 coses,5 cases were treated by LC combined with intraoperative endoscopic sphincterotomy and other 2 cases were converted into open operation.No Severe complication occurred. Conclusions Nearly all cases with stones impacted at cystic duct can receive LC by removing impacted stones through incising the cystic duct and intraoperative cholangiography.If there are any stones in common bile duct,they can be removed by endoscopic sphincterotomy.

Objective To investigate the protective effect of IL-37 on hepatocyte injury against hypoxia/reoxygenation (H/R) by promoting polarization of M2-type macrophages and its molecular mechanisms.Methods Real-time fluorescent quantitative PCR (qRT-PCR) and Western blot were used to detect the levels of IL-37 mRNA and protein in human monocyte-macrophage THP-1 cells with different polarizations.The lentivirus with IL-37 gene was infected into THP-1 cells.The levels of CD206,CD86,ARG1 and iNOS mRNA was detected by qRT-PCR.The levels of CD163 and CD86 protein was detected by flow cytometric analysis.THP-1 cells and L02 cells were co-cultured by Transwell and treated with H/R.The survival rate and apoptotic rate of L02 cells were detected.The levels of alanine transaminase (ALT) and aspartate aminotransferase (AST) in culture medium were measured.The levels of STAT6 and its phosphorylation in THP-1 cells were detected by Western blot.Results The levels of IL-37 mRNA and protein were up-regulated in M2-type macrophages.IL-37 promoted the polarization of M2-type macropahges.M2-type macrophages induced by IL-37 were cocultured with L02 cells,the survival rate was significantly increased by H/R treatment (P =0.015),while the apoptotic rate,ALT level and AST level were significantly decreased (P＜0.001).The level of phosphorylated STAT6 in THP-1 cells overexpressing IL-37 was up-regulated (P ＜ 0.01).Conclusions IL-37 can induce polarization of M2-type macrophages and protect hepatocyte injury against H/R.Its mechanism may be related to STAT6 signal pathways.

Microcapsule technology is a high and new technology under the focused research and development in 21st century, which has been used in many research fields. However, its researches on preparation field should be paid more attention to, especially the research direction of TCM drug delivery system, which is of great value for development and application. This article reviewed the application progress of microcapsule technology in pharmaceutic preparation field, and discussed the strategic significance of the researches on TCM microcapsule preparation, with a purpose to provide research ideas and paths for the researches on the new formulations of traditional Chinese medicine.

Objective To express and identify of Toll-like receptor 4 (TLR4) extracellular fragment in Pichia pastoris and to provide basis for the related researches. Methods The human TLR4 extracellular cDNA fragment was amplified by PCR and after confirming by DNA sequence analysis, was inserted into the Pichia pastoris expression vector pPIC9K containing AOX1 promoter and lead sequence of ? factor gene to form a pPIC9K/TLR4 extracellular recombinant expression plasmid. The recombinant plasmid was transformed into GS115 and high copies transformants were screened with G418. After being identified by colony PCR, transformants were cultured in flasks and induced by 1% methanol. The expression products were analyzed by SDS-PAGE and were identified by Western blot analysis. Results The sequencing showed that TLR4 extracellular gene was identical to that in Genbank. The analysis for the recombinant plasmid DNA digested by enzyme demonstrated that the recombinant expression plasmids of pPIC9K/TLR4 extracellular were constructed successfully. After screening by G418, 200 high-copy-number of transformants were acquired. The special TLR4 extracellular gene segment was obtained by identification with colony PCR, which showed that TLR4 extracellular gene of 5 clones were inserted into Pichia pastoris. After methanol induction for 4 d, the expression proteins came up to 50.35% of total proteins in medium supernatant as shown by SDS-PAGE. Western blot analysis proved that the expression protein could bind to TLR4 monoclonal antibody specifically. Conclusion TLR4 extracellular protein is expressed and identified successfully by Pichia pastoris system, which can provide basis for the researches of further screening of high expression colony, protein purification, and functional research.

Objective To investigate the relationship of the metastasis-associated genes and its copy numbers variation in the highly metastatic human epithelial ovarian cancer cell line HO-8910PM. Methods The differentially expressed genes and its copy number variation between HO-8910PM cell line and normal ovarian tissues was detected by human genome UI33A 2.0 gene chip and human mapping 10K array 2.0 gene chip, and the data was analyzed by bioinformatics. Some of metastasis-associated genes were validated the results of single nucleotide polymorphism (SNP) and cDNA chips by fluorescence in situ hybridization (FISH) and real-time quantitative PCR. Results Integrate analysis of two gene chips data showed that there were 385 differentially expressed genes in the same and 379 SNP positional point (6 of them, included 2 genes) between HO-8910PM cell line and normal ovarian tissues, these copy number amplification of 379 SNP positional point of chromosome were ≥3, which had 240, deletion ≤ 1 had 139. Chromosome location analysis showed that there were 385 differentially expressed genes located at all chromosomes, and 261 of them ( 67.8%, 261/385 ) located at 10 chromosomes, included that 34 (8.8% ), 33 ( 8.6% ), 28 (7.3%), 27 (7.0%), 25 (6.5%), 24 (6.2%) of them located at chromosome 3, 2, 9, 10, 1 and 11 respectively, and 23 (6.0%) of them at chromosome 6 and 12 each, 22 (5.7%) of them at chromosome 4 and 5 each. For the function of differentially expressed genes, the results showed that 99 (25.7% ) genes belonged to the family of enzymes and their regulators, 54 ( 14.0% ) genes associated with signal transduction, 50 (13.0%) genes associated with nucleic acid binding, and 36 (9.4%) genes associated with protein binding. Conclusion We have demonstrated that there are 4 kinds of differentially expressed genes related to metastasis of ovarian cancer, which belonged to the families enzyme and its regulator, nucleic acid binding, signal transduction and protein binding, and located at chromosome 1, 2, 3, 4, 5, 6, 9, 10, 11 and 12.

Objective To explore the characteristic manifestations of breast malignant isodense masses using mammography. Methods 121 breast isodense masses with pathological confirmations were collected.Compared indicators with pathological findings were the tumor size,shape,edge,structure changes around the mass,axillary lymph nodes,skin changes,nipple changes,suspi-cious malignant calcification.χ2 test and Logistic regression analysis were used to evaluate the diagnostic values of each indicator.Re-sults In the 121 isodense masses,73 were benign,48 were malignant tumors.According to χ2 test analysis,factors impacted the nature of mass including:tumor morphology (χ2 =14.376,P =0.002),tumor edge (χ2 = 21.555,P =0.000),structure twisted surrounding the mass (χ2 =26.939,P =0.000),axillary lymph nodes (χ2 =1 6.285,P =0.000),skin thickening (χ2 =4.698,P =0.030).According to multivariate binary Logistic regression analysis,risk factors of the Logistic regression model included:edge infiltrating of mass (P =0.014),structure twisted surrounding the mass (P =0.003),axillary lymph nodes (P =0.026).Conclusion The characteristic manifestations of breast malignant isodense masses include edge infiltrating,structure twisted surrounding the mass,axillary lymph nodes.These manifestations are of great significance in differential diagnosis of breast isodense masses.

OBJECTIVE: To assess the diagnostic value of CT peritoneography for peritoneal complications of continuous ambulatory peritoneal dialysis (CAPD). METHODS: CT peritoneography in 11 CAPD patients with clinically suspected dialysis-related complications was prospectively studied. The CAPD patients were all treated in Renji Hospital from 2005 to 2007. CT images were reviewed according to the evidence of peritoneal leaks, hernias, loculate pleural fluid collections, and adhesions. RESULTS: Abnormal findings were found in 9 of 11 CAPD cases including inguinal hernias (3 cases), umbilical hernia (1 case), hydrocele (1 case), leaks in catheter tunnel (2 cases), and peritoneal adhesions (2 cases). Parts of them were confirmed by surgical operation. CONCLUSION: CT peritoneography is useful for the evaluation of complications related to CAPD, and it offers excellent tissue contrast and multiplanar imaging for assessment of the complications.

Objective To investigate the pathogenetic role of endogenous angiotensin Ⅱ (AngⅡ) in the mechanism of stress ulcer in obstructive jaundice rats and to detect the effect of angiotensin converting enzyme inhibitor (ACEI) on stress ulcer in obstructive jaundice rats. Methods After common bile duct ligation (CBDL) in Wistar rats, the content of plasma and gastric mucosal AngⅡ, gastric mucosal blood flow (GMBF) and gastric mucosal damage were measured, and the relationship among them was analyzed.Results The plasma AngⅡ contents increased much more significantly at 1, 3, 7 and 14 days following CBDL than those in non-CBDL rats (P<0.05, <0.01, <0.01 and <0.01, respectively). Within 120 minutes following cold-restraint stress, plasma and gastric mucosal AngⅡ contents were elevated, GMBF decreased, and ulcer index and gastric mucosal damage increased more significantly than those in non-cold-restraint stress rats (P<0.05, <0.05, <0.01, <0.01 and <0.05, respectively). Administration of an ACEI, enalaprili, to CBDL rats (5 mg*kg-1*day-1, orally for two days) before stress reduced both the plasma and gastric mucosal AngⅡ levels, inhibited the decrease of GMBF and decreased ulcer index and gastric mucosal damage (P<0.001, <0.01, <0.01, <0.01 and <0.05, respectively).Conclusion The endogenous AngⅡ plays a significant pathogenetic role in the development of stress ulcer in obstructive jaundice rats, and ACEI may prevent stress ulcer.

AIMTo determine the feasibility of establishing the heterologous expression model of human- serotonin transporter(hSERT or 5-HTT).

METHODScRNA of SERT was transcribed from cDNA, which was cloned in the pOTV vector. Each oocyte of mature xenopus laevis was injected with transcribed cRNA in vivo and incubated at room temperature for 4-9 days. Recording the current induced by 5-HT with voltage clamp technique tested the function of the expressed 5-HT transporter.

RESULTSThe transporter current could be observed in Ringer's solution containing 5-HT, and the 5-HT induced current were concentration-dependent. Norepinephrine and dopamine could not induce the transporter current while the 5-HT induced current could be specifically inhibited by 5-HTT blocker, desipramine.

CONCLUSIONThe results demonstrate that the heterologous expression product in xenopus laevis oocytes is human 5-HT transporter.

Animals ; Carrier Proteins ; genetics ; DNA, Complementary ; genetics ; Female ; Gene Expression ; Models, Animal ; Oocytes ; metabolism ; RNA, Messenger ; genetics ; Serotonin ; metabolism ; Serotonin Plasma Membrane Transport Proteins ; biosynthesis ; genetics ; Xenopus laevis

Objective To construct the care needs assessment index for institutional elderly. Methods A total of 30 experts were consulted by the Delphi method. The indexes were selected and identified according to the inquiry results. Results The response rates of three expert consultation rounds were 93.33％(28/30), 85.71％(24/28), and 79.17％(19/24) respectively. The authority coefficients of the three rounds were all above 0.80. The assessment index includes 4 first level indicators, 13 secondary level indicators, and 48 third level indicators. Weighting results indicated that in terms of the elderly in institutional care, the most important domain of care needs was physical function, followed by ability of activity. At the same time, psychological function and social function in the elderly cannot be ignored. Conclusions The study obtained key elements that should be included in a comprehensive care needs assessment index of the institutional elderly, which laid a solid foundation for further investigation on the formation of specific assessment tools for care needs of the elderly in institutional care.