Objective To discuss the value of intraoperative cholangiography during laparoscopic (cholecystectomy)(LC). Methods Retrospective analysis was made on the clinical data of 398 patients (undergoing) intraoperative cholangiography during LC from June 1996 to Dec 2004. Results Forty cases of common bile duct stone, 3 cases of cystic duct stone, 53 cases of anomalous bile duct, and 6 cases of bile duct injury were detected. All patients were treated accordingly, and none died. Conlusion The clinical use of cholangiography during LC can help to markedly reduce the incidence of residual bile stones and promptly detect bile duct injury and other serious complications.

Objective To detect the serum levels of epidermal growth factor（EGF）in rheumatoid arthritis（RA）patients,and explore the possible role of EGF in RA.Methods The serum level of EGF was measured by enzyme-linked immunosorbent assay（ELISA）in 38 RA patients diagnosed according to dianostic criteria for RA by American Rheumatism Association and 20 healthy controls.Rheumatoid factor（RF）,erythrocyte sedimentation rate（ESR）and C-creative protein（CRP）were also measured in these 38 RA patients.The relation beween the serum level changes of EGF and the corresponding clinical indexes in the two groups was statistically analyzed.Results Compared with the healthy individuals,the serum EGF levels of RA patients increased significantly（P 0.05）.Compared with simple bone rarefaction patients,the serum levels of EGF increased significantly in the bone erosion patients（P 0.05）.The serum levels of EGF in RA patients had no correlation with RF,ESR and CRP levels（P 0.05）.Conclusion The evident increase of serum EGF level in RA patients,especially in bone erosion patients,suggests that EGF may partly play a role in the pathogenesis of RA.

Objective: TO study the change of substance P(SP) level in Plasma and its relationship with ankylosing spondylitis(AS).Method: Radioimmunoassay. Results: The author observed that the level of SP in Plasma of AS is singificantly higher than that of healthy controls. SP level in AS patients during active stage is higher than in those during inactive stage. Conclusion: Increased SP may play an importantrole in the pathogenesis Of AS.

Objective To investigate the relationship between acute biliary pancreatitis (ABP) and anomalous pancreaticobiliary duetal union (APBDU). Methods 131 patients with ABP were enrolled to test the serum total bilirubin (TB), alanine amintransferase (ALT), aspartate amintransferase (AST), alkaline phosphates (ALP), γ-glutamyl transferase (γ-GT). All the patients received medical treatment, and then these tests were performed again. Thereafter, all the patients underwent selective surgery and intra-operative cholangiography was performed to observe the pancreaticobiliary duetal union. Results 27 patients (20.6%) with APBDU were found in 131 patients. Among them, 8 cases (29.6%) was B-P subtype (TypeⅠ), 16 cases (59.3%) was P-B subtype (TypeⅡ) , and the remaining 3 cases was mixed subtype (TypeⅢ). A significant decrease of ALT, AST, ALP, γ-GT after non-surgical treatment in both group of APBDU and NAPBDU was noted (P＜0.05). The serum levels of ALT, AST,γ-GT in APBDU patients were (71.81± 23.19) U/L, (47.85±27.87) U/L, (52.86±31.49) U/L, respectively; and in NAPBDU patients were (51.96±15.40) U/L, (40.77±16.58) U/L, (34.86±26.47) U/L. The difference was statistically significant (P＜0.05). Condusions APBDU is an important etiology of ABP.

Objective To investigate the effect of T lymphocyte proliferation,Th1 and Th2 cytokines on T cells from systemic lupus erythematosus (SLE)patients by blocking 4-1BB/4-1BBL.Methods The proliferation of T cells from 30 SLE patients and 20 normal controls was detected by MTT and the levels of patients stimulated with anti-CD3 McAb were significantly higher than those of T cells without stimulation(P＜IFN-γand IL-4 in SLE were significantly higher than those of normal controls(P＜0.01).There were more ex-pression of IFN-γ and IL-4 in supernatant of T cells from SLE after stimulated with anti-CD3 McAb(P＜0.01).However,the production of IFN-γ and IL-4 was inhibited by anti-4-1BB McAb(P＜O.05),and especially the level of IL-4 was markedly decreased.Conclusion Blocking 4-1BB/4-IBBL can significantlv inhibit the abnormal activation of T cells and the secretion of Thl and Th2 cytokines of SLE.

AIM: To investigate the effect of single immunoglobin IL-1 receptor related protein (SIGIRR) on damage of alveolar epithelial cells in acute lung injury induced by lipopolysaccharide. METHODS: The acute alveolar epithelial cell injury model was constructed by stimulation of A549 cells with LPS. In order to over-express SIGIRR, the A549 cells were transferred with eukaryotic expression vector containing full length SIGIRR cDNA. The transcriptional activity of NF-κB was measured by dual-luciferase reporter assay system. The concentrations of IL-1β, TNF- α and IL-6 were detected by ELISA. The levels of these inflammatory factors between the transfected cells and untransfected cells were compared. RESULTS: The over-expression of SIGIRR inhibited the transcriptional activity of NF-κB. The increases in IL-1β, TNF-α and IL-6 concentrations in alveolar epithelial cells induced by LPS were observed. CONCLUSION: SIGIRR in alveolar epithelial cells inhibits TLR4 signals triggered by LPS and attenuates the inflammatory reactions in alveolar epithelial cells, which plays a protective role against the acute damage of the alveolar epithelial cells.

Objective To evalue the ability of detecting the resistance of cefoxitin-sensitive,penicillin-resistant Staphylococcus by different methods and analyze the antibiotic susceptibility spectrum of coagulase-negative Staphylococcus which are non-mecA-mediated oxacillin resistance. Methods All the isolates were collected from Huashan hospital between 2007 and 2009. The isolates were recovered from various clinical sources, including respiratory tract, urine, secretion and sterile fluids samples. The oxacillin susceptibility of Staphylococcus aureus was determined by cefoxitin disk diffusion test, cefoxitin MIC test,oxacillin disk diffusion test and oxacillin MIC test Likewise, the oxacillin susceptibility of coagulasenegative Staphylococcus was determined by cefoxitin disk diffusion test and oxacillin MIC test. All the isolates with sensitive to cefoxitin were screened for the mec A gene by PCR Finally, the MIC of non-mecA-mediated oxacillin-resistant Staphylococcus were determined. Results Among 255 cefoxitin disk diffusion test sensitive and penicillin-resistant Staphylococcus aureus, 6 isolates were intermediated to oxacillin and 4 were resistant by oxacillin disk diffusion test, but all the isolates were sensitive by the cefoxitin disk diffusion test,cefoxitin MIC test and oxacillin MIC test. Among 75 cefoxitin disk diffusion test sensitive and penicillin-resistant coagulase-negative Staphylococcus, 16 isolates were resistant to oxacillin by oxacillin MIC method and 4 carried mecA gene. Among 12 non-mecA-mediated oxacillin-resistant Staphylococcus, the susceptible isolates of gentamicin is 10, clindamycin is 8, ciprofloxacin is 11, erythrornycin is 6, trimethoprim/sulfamethoxazo]e is 11 ,and cephalosporins, teicoplaninl, vancomycin, piperacillin/tazobactam, tetracycline are all 12. Conclusions The cefoxitin disk diffusion test can reliably predict mecA-mediated oxacillin resistant Staphylococcus aureus. It would be best to combine cefoxitin disk diffusion test and oxacillin MIC test to improve accuracy of detection of mecA-mediated oxacillin resistant coagulase-negative Staphylococcus.Furthermore, infections due to the non-mecA-mediated oxacillin resistant coagulase-negative Staphylococcus can be treated by penicillinase-stable penicillins, β-lactam/β-lactam inhibitor combinations, relevant cephems and carbapenems.

Objective Cyclin D1 gene plays a significant role in regulating cell cycle progression.It is reported that over-expression of cyclin D1 gene is intimately associated with origination,development and prognosis of tumor and is associated with tumor cells resistance to chemotherapy drug.Suppression of cyclin D1 protein expression leads to cellular chemosensitization.This study was to determine whether this effect also existed in chronic leukemia cell line K562 by inhibiting the expression of cyclin D1 protein through RNA interference.Methods Plasmid vectors expressing small hairpin RNA (shRNA) targeting at cyclin D1 gene were constructed and transfected into K562 cells by chitosan.Cyclin D1 protein was examined using Western Blotting analysis.The cell cycle and apoptosis were determined by flow cytometry.Cellular chemosensitization was evaluated by MTY assay.Results Expression of cyclin D1 protein was markedly down-regulated after transfection with pshRNA-419 and pshRNA-575 at 48 h.Down-regulation of cyclin D1 protein could affect the redistribution of cell cycle,induce apoptosis of K562 cells,decrease 50％inhibitoryconcentration (IC50) of adriamycin and enhance cellular chemosensitization.But there had no above biological effects observed after transfection with blank vector and control vector of m-pshRNA-790 at 48 h.ConclusionK562 cells could be chemosensitized by the down-regulation of cyclin D1 expression through RNA interference.

Objective To establish a system for detecting integration frequency of antibiotic resist-ante integron.Methods We cloned integron and aadA2 gene cassette into different sites of plasmid pACYC 184,and the plasmid was transformed into E.coli BL21(DE3)containing plasmid overexpressing integrase.The positive clone was cultured overnight and then was spread on LB agar plate with or without streptomycin respectively,and with appropriate amount of bacteria.Clones after cultured overnight were counted to detect the integration frequency.Meanwhile we used positive clones in LB agar plate containing streptomycin as templates to carry out PCR.The purified PCR products were sequenced to identify the integration sites.Re-suits The integration frequency of integron capturing aadA2 gene cassette in BL21(DE3) host was 1.1 x 10-3 mainly at attI site.Conclusion This system can be used to detect integration frequency.

AIM: To compare the therapeutic effects of transplantation of human umbilical cord mesenchymal stem cells (hUCMSC) through different ways on diabetic mice.METHODS: hUCMSCs were labeled with enhanced green fluorescent protein (EGFP) and luciferase (Luc) reporter gene, and then the cells were transplanted into the diabetic mice through pancreas or tail vein to monitor the migration of the hUCMSCs in vivo.The pathological changes of pancreas tissue sections were determined by HE staining.Weight and blood glucose of the mice were measured dynamically.To compare the therapeutic effects, serum insulin levels were analyzed and glucose tolerance test were also performed.RESULTS: In vivo bioluminescence imaging results showed that the hUCMSCs transplanted into pancreatic capsule was mainly located in the pancreas while the hUCMSCs transplanted through vein tail injection was mainly located in the lung.HE staining illustrated that islet cells presented distinctive boundary and no infiltration of inflammatory cells in pancreatic capsule transplantation group was observed, but a little inflammatory cell infiltration and fibrosis formation in tail vein injection group were seen.A significant decrease in blood glucose level and a significant increase in serum insulin level in pancreas transplantation group were showed as compared with vein tail injection group.CONCLUSION: Transplantation of hUCMSCs through different approaches demonstrates different effects.The transplantation of hUCMSCs into pancreatic capsule is more effective on hyperglycemia reversion, insulin secretion and improvement of beta-cell function than that through tail vein.