BACKGROUND: Calcium pyruvate has great effect on reducing bodymass, and chitosan can regulate immunity system, promote bone growth, decrease blood sugar and adjust blood lipid. Calcium pyruvate and chi tosan capsule (CCC) was made by calcium pyruvate and chitosan combined with Chinese medicine, which can be used both as food and medicine. However, its effect still needs further observation. OBJECTIVE: To investigate the weight-reducing effect of CCC on obesity rats, and explore its mechanism.DESIGN: A randomized controlled trial.SETTING: Teaching and Research Section of Preventive Medicine, Department of Public Health, Xi'an Medical College, and Department of Public Health, Medical School of Xi' an Jiaotong University.MATERIALS: The experiment was conducted in the Department of Public Health, Medical School of Xi'an Jiaotong University from April to July 2001. Totally 60 healthy male weaning SD rats with body mass of 50-80 g,were provided by the Animal Center of Xi'an Jiaotong University. CCC,mainly composed of calcium pyruvate and chitosan, mixed with Chinese medicine extract after prepared with distilled water, was made by the author.METHODS: ①Grouping and modeling: The 60 rats were randomly divided into 5 groups: blank control group, model control group, high, middle and low dose CCC supplement groups, respectively with 12 rats in each group. The rats in the blank control group were fed w ith basal diet; the other groups were fed with high fat and nutrition diet to establish rat models of nutritional obesity. ②Administration: The rats in the blank and model control groups were given suspension mixed with starch matrix in capsule and distilled water at same dose. The rats in the high, middle and low dose CCC groups were intragastrically infused with 3, 1.5 and 0.75 g/kg CCC, once aily for 30 days. ③Detection: Body mass, body length and wet weight of fat tissue were measured before and after administration to calculate the obese index and ratio of fat weight/body mass. The adipocyte number and adipocyte size were observed by 400-fold microscope and ocular micrometer, respectively. The detection of serum triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C) and serum leptin levels were performed according to the leptin radioimmunity analyzer kit.MAIN OUTCOME MEASURES: ①Body mass, body length and obesity index before and after administration. ②Wet weight of fat, fat/body mass,adipocyte number and size. ③TG, TC, HDL-C and serum leptin.RESULTS: All the 60 rats were involved in the result analysis. Before administration, the body mass of the blank control group was obviously lower than other 4 groups (P ＜ 0.01), and there was no significant difference among the 4 groups (P ＞ 0.05). After administration, the mean body mass,obesity index, wet weight of fat tissue, ratio of fat/body mass and leptin level of the high, middle and low CCC groups were lower than the model group (P ＜ 0.05 or 0.01), and the adipocyte size were significantly smaller than those in the model group (P ＜ 0.05 or 0.01), but the adipocyte number was more than the model group (P ＜ 0.05 or 0.01). Meanwhile, level of TG and TC of the high and middle dose CCC groups were lower than the model group (P ＜ 0.05 or 0.01), there was no significant difference in the body length and HDL-C of each group.CONCLUSION: ①CCC shows evident weight-reducing effect on rats with nutritional obesity, which is not related to the ability of CCC to enhance the serum leptin level. ②CCC can also lower the blood lipid level.

Objective To investigate the effectiveness of choledochoscopy and duodenoscopy in the treatment of residual calculus of the intrahepatic bile duct. Methods Clinical data of 573 cases of choledochoscopy and 36 cases of duodenoscopy from February 1985 to June 2004 were analyzed. Results Stones were completely removed under choledochoscope by way of T-tube sinus tract in 96.02% of patients (507/528) and by way of subcutaneous blind loop in 84.62% of patients (22/26). Percutaneous transhepatic choledochoscope placement was successfully conducted in 84.21% of patients (16/19) and stones were completely removed in 78.95% of the patients (15/19). The success rate of duodenoscopic cholangiography was 91.67% (33/36) and stones were completely removed under duodenoscope in 80.56% of patients (29/36). Conclusions Treatment of residual and recurrent calculus of the intrahepatic bile duct by choledochoscopy and duodenoscopy is effective and convenient.

Objective To analyze the MRI data of misdiagnosed and missed diagnosed of breast lesions and their histopathological features.Methods Data from 241 breast lesions within 121 patients were recruited in this study.The data included MRI images,uhrasounds and X-ray images were retrospectively interpreted by two radiologist and each lesion was assessed according to the BI-RADS classification.The pathologic features of miss or error diagnosed lesions on MRI were analyzed.Results In 241 breast lesions (malignance 120,bcnign 121),4 lcsions were miss diagnosed on MRI.Thcy were 2 intraductal papillomatosis and 2 fibroadenoma.All was benign.Twenty three lesions were misdiagnosed on MRI.Sixteen were overestimation,including 3 chronic inflammations,3 sclerosing adenosis,2 fibroadenoma,4 fibrocystic changes with or without atypical ductal hyperplasia (ADH),2 intraductal papilloma,1 infiltration of pectoralis major muscle and 1 axillary lymphnode metastasis.Meanwhile,there were 7 lesions were underestimation.These lesions included 2 invasive ductal carcinomas,1 mucinous adenocarcinoma,2 DCIS and 1 blunt duct adenosis with ADH and focal cancerous,1 inflammatory breast cancer underwent chemotherapy.The sensitivity and specificity and accuracy of breast MRI were 95.83 ％ (115/120),72.73 ％ (88/121),84.23 ％ (203/241),respectively.MRI findings had no difference with respect to mammogram or ultrasound was 75.10 ％ (181/241).Conclusion MRI misdiagnosis and missed often occurs in smaller breast lesions,morphologic and hemodynamic malignant manifestation atypical,especially intraductal lesions.MRI diagnosis should be combined with physical examination,X-ray mammogram and ultrasound to improve diagnostic accuracy and reduce missed diagnosis.

Objective To determine the best method to detect thermo-acidophilic bacteria in apple juice concentrate. Methods L 16 (4 5) orthogonal test was applied to analyse the four factors, including culture medium, ways of heat shock, dilution degrees of AJC and pH value. The results were comprehensively scored according to the number of colonies. Results The isolated bacteria were polymorphous. A better method was as follows: after being diluted to 1∶10, the AJC was submitted to a heat shock at 80 ℃ for 10 min, and then spread on k agar with a pH value of 3.5. Samples were incubated at 45 ℃ for 3 days, and then enumerated. Conclusion This method can considerably improve the sensitivity of detecting thermo-acidophilic bacteria in apple juice concentrate.

Objective To observe the effect of astragalus polysaccharides ( APS) on glucose homeostasis regulation and focus on glycogen synthase kinase 3 beta (GSK3 beta) activity and subcellular localization (nuclear translocation).Methods HepG2 human hepatoma cells were cultured in vitro and treated with high glucose of different concentrations (30, 40 mM) to induce hepatocyte endoplasmic reticulum stress model, then acquire optimum operating concentration.The HepG2 cells were treated with APS of different concentrations (50, 100, 200, 400 μg/mL) to select the most effective concentration.The HepG2 cells were divided into seven groups with different treatment: negative control group (C), positive control group (Tm), 30 mM high glucose-induced group (G30), 45 mM high glucose-induced group (G45), negative control+APS group (CA), positive control+APS group ( TA) and high glucose-induced+APS group ( GA).Effect of APS at different concentrations on proliferation activity of HepG2 cells were detected by MTT assay, transcription and shear levels of XBPlmRNA in HepG2 cells by quantitative real-time PCR, and phosphorylation levels of GSK3βin cytoplasm and nucleus by immunoblotting techniques.Results The optimum operating glucose concentration was 30 mM.The most effective APS concentration was 200μg/mL.The transcription and shear levels of XBPlmRNA in HepG2 cells of GA group were lower than those of G30 group ( P<0.05), respectively, but there were no significant differences between TA and Tm group.The phosphorylation levels of GSK3βin cytoplasm and nucleus of GA group were higher than those of G group(P<0.05), respectively, but there were no significant differences between TA and Tm group. Conclusion APS could improve hepatic steatosis, and its mechanism might be that APS inhibits the activity and nuclear localization of GSK3β, then alleviate endoplasmic reticulum stress.

Objective To investigate the effect of reinforced Decoction of Angelicae Sinensis for enriching blood (RDAEB) on the immunity of immunosuppressed mice induced by cyclophosphamide (Cy). Methods Mice were given RDAEB through stomach perfusion for 10 d(50 mg/d). Then, RBC-C3bR rate,RBC-IC rate( as the indexes of erythrocyte immunity)and E-rosette forming rate,acidic α-naphthyl acetate esterase positive rate, lymphocyte transformation rate (as the indexes of cellular immunity) of mice were tested.Results RBC-C3Br rate, RBC-IC rate,E-rosette forming rate, acidic α-naphthyl acetate esterase positive rate and lymphocyte transformation rate in the Cy-RDAEB group were markedly higher than those in the Cy group (P＜0. 01 ),and returned to the levels of normal group. Conclusion RDAEB is effective in recovering and enhancing cellular and erythrocyte immunity of immunosuppressed mice.

ObjectiveTo explore the relationship between the negative co-inhibitor programmed death-1 ( PD-1 ) and the pathogenesis of myasthenia gravis ( MG), by detecting the expression of PD-1 and programmed death ligand-1 ( PD-L1 ) on peripheral blood mononuclear cells (PBMCs) and soluble PD-1 (sPD-1) in plasma from myasthenia gravis patients. MethodsPeripheral blood samples were collected from 45 MG patients and 33 healthy persons without prednisone or other immunodepressant treatment during the half year ahead of withdrawal.The expression of PD-1 and PD-L1 on PBMCs were detected using immuno-fluorescence labeling and flow cytometry, and the concentrations of sPD-1 in plasma were measured using an ELISA kit. Results(1) The proportion of CD4+ PD-1 + T cells, as well as CD14+ PD-L1 +monocytes of the MG group was higher than that of the control group. There were no significant differences in the proportion of CD4+ PD-1 + T cells or CD14+ PD-L1 + monocytes in the MG sub-groups between different genders or MG types. While the proportion of CD4+ PD-1 + T cells of the late-onset MG (age ≥40) group was higher than that of the early-onset MG group (age ＜40). And it was higher in the MG patients with thymoma or thymus hyperplasia than that from the MG patients with normal thymus. The proportion of CD14+ PD-L1 +monocytes from the MG patients with thymoma or thymus hyperplasia group decreased obviously compared with that of the patients with normal thymus group; but no difference could be found between the late-onset group and early-onset group. (2)The concentration of sPD-1 in the plasma from the group of MG patients was(6. 92 ±0. 72) ng/ml,which was higher than that of the healthy control group ( (3.28 ±0. 42) ng/ml),even more, it was significantly higher in the early-onset MG group than that of the late-onset MG group,there was a negative correlation( r =-0. 526, P =0. 000) between the age of onset and the concentration of sPD-1. ConclusionsThe increased expressions of PD-1 on CD4+ T cells and PD-L1 on CD14+ monocytes in MG patients suggested the involvement of the couple of molecules in the pathogenesis of MG.Higher concentration of soluble PD-1 in the plasma of patients with MG suggested that it might disturb the ligation of PD-1 and PD-L1 on T cells and antigen presenting cells, which might result in the abnormal transportation of the negative modulating signal, and accelerate the pathological progress of MG.

Objective To investigate the role and mechanism of B7-H4, a negative costimulatory molecule, in mediating the immunomodulatory effects of mesenchymal stem cells C3H10T1/2 (C3H10) on T cell polarization. Methods The lentiviral vectors that carried the shRNA targeting mouse B7-H4 were transfected into mouse mesenchymal stem cells (C3H10-B7-H4). The cells were co-cultured with PHA-acti-vated mice spleen lymphocytes before and after the transfection. ELISA was performed to detect the concen-trations of cytokines in supernatants of cell culture in order to elucidate the effects of B7-H4 expressed by C3H10 on T cell polarization. A mouse model of experimental allergic encephalitis (EAE) was established. Fifty C57BL/6 mice were divided into five groups including control group, EAE group, C3H10 group (injec-ting EAE mice with C3H10 cells), C3H10-NC group ( injecting EAE mice with C3H10-NC cells) and C3H10-B7-H4 group (injecting EAE mice with C3H10-B7-H4 cells). ELISA was performed to detect the soluble form of IL-2, IL-17, IFN-γ and IL-4 in plasma samples. Results Knocking down the B7-H4 gene with shRNA significantly decreased the expression of B7-H4 on C3H10 cells, which weakened the inhibitory effects of C3H10 cells on the secretion of IL-2, IL-17 and IFN-γ by spleen lymphocytes. The therapeutic effects of C3H10-B7-H4 cells on mice with EAE were weakened after silencing the B7-H4 gene expression, which was manifested as higher nerve function score and earlier onset and bring forwarded peak time of EAE than those of the C3H10 group. Treating EAE mice with C3H10-B7-H4 cells was less efficient in inhibiting the expression of IL-2, IL-17 and IFN-γin plasma. However, knocking down the B7-H4 gene had no signif-icant effect on the expression of IL-4 in terms of treating EAE with C3H10 cells. Conclusion The co-inhib-itor molecule B7-H4 expressed on C3H10 cells mediated the treatment of EAE with C3H10 cells by regula-ting Th1 and Th17 effector T cells.

Objective To investigate the role of circulating neutrophil extracellular trap (NET) levels in the postoperative formation of deep vein thrombosis (DVT) in patients undergoing total knee arthroplasty (TKA). Methods Circulating NET levels were measured preoperatively and on postoperative days 1, 3, and 7 in 30 patients diagnosed with DVT by venography after TKA and 30 controls from May 2014 to July 2015. Results In patients with DVT, the mean circulating NET levels were significantly higher on postoperative days 1 and 3 compared with those in the non?DVT group (postoperative day 1, 1.598 ± 0.067 vs. 1.212 ± 0.037, t=7.514, P=0.000;postoperative day 3, 1.305±0.044 vs. 1.167±0.032, t=2.675, P=0.015). ROC curve analysis revealed the inferior sensitivity and specificity of NET levels on postoperative day 3 compared with postoperative day 1. Area under the curve (AUC) postoperative day 1 was 0.828 (95%CI:0.722, 0.933, P=0.000);AUC postoperative day 3 was 0.677 (95%CI:0.541, 0.814, P=0.018). The cutoff point for NET levels on postoperative day 1 was 1.294, with a sensitivity of 80%, a specificity of 80%, a posi?tive predictive value of 80%, and a negative predictive value of 80%. After Logistic regression, the NET level on postoperative day 1 was considered an independent predictor of DVT (OR:24.08, 95%CI:4.94, 117.28, P=0.000). Conclusion High circulating NET levels are associated with DVT in patients who undergo TKA. NETs may serve as a potential biological marker to delineate patients undergoing TKA who are most at risk for DVT.

This study aimed to investigate the effects of Sargassum fusiforme polysaccharide (SFPS I, II, and III) on the apoptosis and regulation of human erythroleukemia (HEL) cells. The effect of different doses of SFPS on HEL cell growth was detected using the Cell Counting Kit-8 method, and apoptosis was detected by Hoechst staining. Cell cycle distribution and apoptosis were detected using flow cytometry. Expression of the cell cycle gene, p53, antiapoptotic genes, Bcl-xL and Bcl-2, and pro-apoptotic genes, Bax, Bad, and Caspase-3, as well as the expression of the corresponding proteins, were detected using real-time quantitative polymerase chain reaction (qPCR) and Western blot. The results showed that SFPS II and III decreased HEL cell viability and induced HEL cell apoptosis. Different concentrations of SFPS (I, II, and III) were detected that induced much less toxic effect in normal human embryonic lung (MRC-5) cells, and SFPS I increased cell proliferation, indicating its favorable selectivity towards cancer cells. The mechanism by which SFPS induced apoptosis was also found to be related to the induction of cell cycle arrest in the G/G phase and the increased expression of apoptosis-related genes and proteins. We concluded that SFPS induces HEL cell apoptosis, possibly via activation of the Caspase pathway, providing the theoretical basis for the development of SFPS-based anti-tumor drug products.