Gastrins ; metabolism ; Gonadal Steroid Hormones ; metabolism ; Growth Hormone ; metabolism ; Hormones ; metabolism ; Humans ; Liver Diseases ; metabolism ; Thyroid Hormones ; metabolism

Objective To determine the role of liver function test in the preoperative diagnosis of concomitant asymptomatic common bile duct (CBD)stone in patients with cholecystolithiasis.Methods A retrospective study was conducted from January 2012 to October 2013 at the Department of Hepatobiliary Surgery,the Affiliated Hospital of Zunyi Medical College on 426 patients who were operated for cholecystolithiasis.According to whether they had abnormal liver function,these patients were divided into the CBD stone group (n =44) and the cholecystolithiasis group (n =382).The values of the different components of liver functions test such as ALT,AST,AKP,GGT,TBIL and DBIL in diagnosing CBD stone were statistically analyzed.Results This study involved 426 patients,159 men and 231 women,with a mean age of (50.96 ± 12.93) years.44 patients with both CBD stone and cholecystolithiasis.The rates of abnormal liver function were 77.27％ (34/44) vs 4.45％ (17/382) in CBD stone group vs cholecystolithiasis group.The difference was significant (x2 =198.54 ; P =0.000).In logistic regression analysis,only elevated serum level of GGT (OR =10.067 ; P =0.007) remained as an independent predictor of CBD stone.On ROC (receiver operating characteristic) curves analysis,the area under the curve was 0.881.With a cut-off point for GGT at 84.5 U/L,there was a sensitivity of 72.2 per cent,specificity of 96.1 per cent,and positive and negative predictive values of 76.3 per cent and 96.2 per cent respectively.Conclusion Our study showed that liver function,especially GGT,was a predictor of CBD stone.

Inteleukin-37 is a member of IL-1 family and originally named as IL-1F7.It has five different subtypes (IL-37a-e).It has been reported that IL-37 suppresses pro-inflammatory cytokines production of a variety of immune cells, and further regulate innate immune response. In addition, IL-37 has protective effects on colitis, arthritis, pancreatitis and other inflammatory diseases that induced by exogenous stimuli. In a word, IL-37 is a novel anti-inflammatory cytokine and plays an important role in a variety of inflammation-related diseases.

Objective To investigate the expression of recombinant IL-37b protein and removal of the endotoxin, and identify its biological activity.Methods The prokaryotic expression vector pET28/IL-37b was constructed and to transform Escherichia coli (E.coli) Rosetta.After induction with IPTG, the recombinant protein was purified through Ni2+-NTA gel column and identified by SDS-PAGE and Coomassie brilliant blue staining.Then, the endotoxin protein was removed and was treated with LPS-stimulated RAW 264.7 cells.The culture supernatant was collected.The expression of IL-6 was detected by ELISA and the biological activity of the protein was identified.Results The recombinant IL-37b with high purity was expressed and the endotoxin produced by prokaryotic expression was reduced, and it was identified to have good biological activity.Conclusions In this study a recombinant IL-37b protein with high biological activity is successfully obtained.

Objective To establish a C57BL/6 mouse model of intestinal infection induced by S.Typhimurium.Methods In order to improve the infectious sensitivity of S.Typhimurium, C57BL/6 mice were intragastrically given 5% (w/v) NaHCO3.Then mice were challenged with S.Typhimurium.The health condition, survival and body weight of mice were observed from day 0 to day 7 after the bacterial infection.The pathological changes were also examined.Results the mice challenged with S.Typhimurium showed decreased body weight and typical clinical signs, including in appetence, piloerection and low survival rate.Macroscopic dissection revealed that intestinal hyperemia and swelling were founded in the mice challenged with S.Typhimurium.Histopathology showed intestinal epithelial and mucosal damages.Conclusions We have successfully established a C57BL/6 mouse model of S.Typhimurium infection.This model may be of crucial significance for studying the biological functions of associated immunological molecules or cytokines in the process of inflammatory bowel disease induced by S.Typhimurium.

Objective Resveratrol can improve nonalcoholic fatty liver disease , but its action mechanisms remain unclear . This study aimed to investigate the protective effect of resveratrol against the free fatty acid ( FFA)-induced apoptosis of human hepatic L02 cells and its possible mechanisms . Methods Human hepatic L02 cells were incubated with FFA and resveratrol for 24 hours.The prepared cells were divided into a blank control , an FFA ( 2 mmol/L) , and a resveratrol group ( 50 μmol/L resveratrol +2 mmol L/FFA).After treatment, we measured the triglyceride (TG), glutathi-one (GSH), and malonaldchyde (MDA) contents and caspase3 ac-tivity in the hepatocytes , determined the apoptosis of the cells by flow cytometry , and detected the protein expression of silent information regulator 1 (SIRT1) by Western blot as well as the mRNA expressions of catalase (CAT), Mn superoxide dismucase (MnSOD), Bcl-2, and Bax by qRT-PCR. Results The TG content and caspase 3 activity in the hepatocytes were significantly increased in the FFA ([3518.±64.2] μmol/L and [5.97 ±0.78] U/g) and the resveratrol group ([201.1 ±60.1] μmol/L and [3.60 ±0.73] U/g) as compared with those of the blank control ([40.2 ±7.4] μmol/L and [2.56 ±0.49] U/g) (both P<0.05), but the caspase3 ac-tivity was markedly decreased in the resveratrol group in comparison with that of the FFA group (P<0.05).Both early and late apop-tosis rates of the hepatocytes were remarkably higher in the FFA ([6.75 ±0.81]%and [8.52 ±0.54]%) and the resveratrol group ([4.94 ±0.44]%and [6.52 ±0.61]%) than those in the blank control ([3.38 ±0.33]% and [2.72 ±0.19]%) ( both P<0.05), with statistically significant differences between the former two groups (P<0.05).The resveratrol group showed significant differences in the GSH content ([100.2 ±8.8] nmol/g), the MDA level ([2.36 ±0.82] mg/g), and the relative expression of SIRT1 (0.84 ±0.04) from the FFA group ([73.8 ±13.1] nmol/g, [3.77 ±0.92] mg/g, and 0.61 ±0.07) and the control ([113.7 ±13.8] nmol/g, [1.85 ±0.41] mg/g, and 0.90 ±0.02) (all P<0.05).The resveratrol group also exhibited statistically significant differences in the relative expressions of the MnSOD , CAT, and Bax genes from the FFA and control groups (P<0.05). Conclusion Resveratrol attenuates FFA-induced apoptosis of human hepatic L 02 cells by activating SIRT1 and reducing the oxidative stress of hepatocytes .

Objective To investigate the molecular epidemiology and risk factors of the expression of the efflux pumps of multidrug resistant Pseudomonas aeruginosa (Pa). Method From April 2014 to October 2014 , clinical features of 100 patients infected with multidrug resistant Pa were retrospectivly analyzed . Results MexAB-OprM was expressed in all multidrug resistant Pa strains (100%). The expression rates of MexEF-OprN and MexCD-OprJin the multidrug resistant Pa strains were 50% and 36%, respectively. The risk factor of the three efflux pumps expression was analyzed and the results were as follows: the use of carbapenems and glycopeptides antibiotics was the risk factor of MexAB-OprM overexpression which was induced. The use of macrolide and carbapenems antibiotics was the risk factor of the induced MexEF-OprN and MexCD-OprJ expression. Invasive procedures, low WBC, anaemia, chronic fundamental disease and hospitalized in ICU ward were the common risk factors of communicable expression or overexpression of the three efflux pumps. Invasive procedures, low WBC, anaemia, chronic fundamental disease and hospitalized in ICU ward were commonly independent risk factors of expression of the three efflux pumps. Conclusion The normalized use of carbapenems and glycopeptides antibiotics , the reduction of unnecessary invasive procedures , the severely grasped standard of hospitalized in ICU ward , curing the fundamental diseases and improving the immunonity played important roles in the prevention of the expression or overexpression of the efflux pumps of multidrug resistant Pa.

Objective To evaluate the clinical effectiveness of 3?dimensional scaffold of typeⅠcollagen based autolo?gous chondrocyte implantation (ACI). Methods Nine patients of knee articular cartilage defect treated with 3?dimensional scaf?fold of type Ⅰcollagen based ACI from January 2013 to March 2014 was analyzed retrospectively, including 6 males and 3 fe?males with an average age of 30 years old. 4 defects located in femoral condyle, 4 in trochlea and 1 in patellae with a mean size of 4.9 ± 2.1cm2 (range, 2.5-10). ACI comprises 2?stage procedure:chondrocytes were first harvested from non?load bearing area of femoral condyle, then chondrocytes expand in vitro for 8-14 days to get enough cells. On second stage, cartilage defects were cov?ered by the grafts and fixed with fibrin albumen glue. All patients received strict rehabilitation protocol. International Knee Docu?mentation Committee (IKDC) scores and Lysh?lm scores were compared pre?operatively and 3, 6, 12 months post?operatively. MR and magnetic resonance observation of cartilage repair tissue (MOCART) scores were analyzed within 3 days, 3, 6, 12 months post?operatively. Results All the patients were followed up. IKDC score was 52.7 ± 6.9 pre?operatively and respectively 71.1 ± 6.6, 83.3±2.9 and 92.0±3.6 3, 6, 12 months post?operatively with significant differences. The Lysh?m score was 55.8±8.7 pre?oper?atively and respectively 74.8±7.0, 84.8±4.8 and 93.1±5.7 3, 6, 12 months post?operatively with significant differences. 8 patients had MRI. The mean MOCART score 3 days, 3, 6, 12 months post?operatively was respectively 43.6±6.0, 47.8±5.8, 57.8±5.8, 64.3± 4.8 and 72.1±4.9 with significant differences. T2 value of transplanted area was 48.7±3.2 12 months post?operatively with no sig?nificant differences compared to normal area. Conclusion Three?dimensional scaffold of typeⅠcollagen based ACI could re?pair knee articular cartilage defect. It may be a good choice for treating articular cartilage defect which shows satisfactory results.

To enhance the specificity of anti-TNF-α single chain Fv antibody (TNF-scFv) to inflamed site, we constructed a bispecific antibody BsDb that targets TNF-α and ED-B-containing fibronectin (B-FN) by covalently linking TNF-scFv and the anti-ED-B scFv L19 at the gene level via a flexible peptide linker deriving from human serum albumin. BsDb was successfully secreted from Pichia pastoris as functional protein, identified by immunoblotting, and purified to homogeneity with affinity chromatography. BsDb retained the immunoreactivity of its original antibodies TNF-scFv and L19, and showed a marked gain in antigen-binding affinity and in TNF-α-neutralizing ability, when compared to TNF-scFv and L19 that were produced in Escherichia coli. In the adjuvant-induced arthritis (AIA) mice model, BsDb showed selective accumulation and retention in the inflamed paws but rapid clearance from blood, resulting in high arthritic paw to blood ratios. These data indicate that BsDb is endowed with high specificity to inflamed site and low toxicity to normal tissues and holds great potential for in vivo application for the targeted therapy of RA and other chronic inflammatory diseases.
Animals ; Antibodies, Bispecific ; biosynthesis ; immunology ; Antibodies, Neutralizing ; biosynthesis ; immunology ; Escherichia coli ; Fibronectins ; chemistry ; immunology ; Humans ; Mice ; Single-Chain Antibodies ; biosynthesis ; immunology ; Tumor Necrosis Factor-alpha ; immunology

In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.