Objective To explore the regulating mechanism of bcl 2 gene in hepatocyte injury caused by obstructive jaundice in rats. Methods Normal rats' and bile duct ligated 7d,14d,21d rats' hepatocytes were isolated by in situ collagenase perfusion and primary culture. (1) bcl 2 mRNA was detected by RT PCR in all cells; (2) After normal rat and bile duct ligated 14d rat hepatocytes were added 100?M GCDC and kept for 24hrs, cells were evaluated by FCM and TUNEL. Results (1) Normal rat hepatocytes did not express bcl 2 by RT PCR technique. bcl 2 was expressed in 7-,14-,21-day BDL rats. (2) After adding 100?M GCDC and keeping for 24hrs, the apoptosis of bile duct ligated rat hepatocytes significantly decreased compared with that of normal rat hepatocytes. Conclusions (1) Bile duct ligated rat hepatocytes expressed bcl 2. (2) Hepatocellular expression of bcl 2 during obstructive jaundice is an adaptive phenomenon to resist apoptosis by bile salts.

Objective To investigate the effects of epigenetic drugs on 2D- and 3D-cultured cholangiocarcinoma cells in vitro.Methods In this study,we have built compact and round TFK-1 spheroid in poly-HEMA coated 96-well plate and determined the effects of epigenetical drugs on 2D and 3D cultured cholangiocarcinoma cells:TFK-1.Viability of 2D and 3D cells model was examined by WST assay and FDA/PI staining. Using methylation-specific PCR analysis,we demonstrated the changes of methylation status of promoters regarding three tumor suppressor genes APC,E-Cadherin,and p16 INK4A.Results The average diameters of compact and round TFK-1 spheroids were in the range of 350-400 μm.The TFK-1 spheroid cells were more resistant to the epigenetic drugs and demonstrated a 11.2155-fold higher IC50 values for hydralazine and valproic acid than the same cells grown as monolayer. Higher doses of epigenetic drugs were needed to reverse the hypermethylation status in 3D cultured cells than 2D cells; however,the parallel dosage - dependent characteristic did not show in the 3D spheroid group.Conclusions Taken together,we established a 3D culture model of human cholangiocarcinoma epithelial spheroid.The 3D spheroid cells are more complex than the 2D monolayer cells and their unique characteristics are able to affect the consequences of epigenetic therapy.The 3D spheroid is a promising model for the research of epigenetic therapy.

Objective To investigate the effects of urinastatin on the production of inflammatory mediators and cytokines of acute pancreatitis(AP), and the effect of treating AP with urinastatin. Methods Serum levels of tumor necrotic factor ?(TNF ?), nitrogen oxide(NO), oxygen free radicals and amylase were determined in AP rats and patients with AP respectively. Effects of urinastatin treatment on the alleration of symptoms, signs and of pancreas in patients with AP were also examined. Results Urinastatin could apparently decrease the serum levels of TNF-?, oxygen free radicals and amylase in AP rats and the patients with AP, also alleviate the symptoms and signs of the patients with AP, and the effective rate of treating AP with urinastatin reached 90 percent. Conclusions Urinastatin, which can inhibit the production of inflammatory mediators and cytokines in AP, is an effective and cheap drug for AP.

Objective To investigate the clinical and laboratory characteristics of anticentromere antibody (ACA)and anti-SSA antibody expressions in patients with Primary Sj(o)gren's Syndrome (PSS). Methods Twelve PSS patients with ACA positive but SSA negative(ACA PSS)and 19 PSS patients with SSA positive but ACA negative(SSA PSS)were enrolled into the study and classified into two groups. We compared the age,laboratory data,occurrence of Raynaud's phenomenon(RP),and histological changes in minor labial salivary glands biopsies of the patients from two group. Results The mean age of the ACA PSS group(68.4 ± 7.9)years was significantly higher than that of the SSA PSS(54. 6 ± 16. 2)years group(P ＜ 0. 05). Serum IgG level of ACA PSS group(17. 89 ±4. 08)g/L was close to the normal range,which was significantly lower than that of SSA PSS(27.90 ±6. 72)(P ＜0. 01). Leukocytopenia was less frequently observed in ACA PSS than in SSA PSS(P ＜ 0. 05),the difference between two groups was statistically significant. We also found more frequent RP in the ACA PSS group than SSA PSS group(P ＜ 0. 05). Conclusions Our data confirm that ACA positive PSS differs from SSA positive PSS at several clinical respects and laboratorial examinations.

BACKGROUND:The non-specific immune suppression method is generaly used for treatment of systemic lupus erythematosus, but poor prognosis, such as infection and high recurrence rate, exists.
OBJECTIVE:To evaluate the therapeutic effect of bone marrow mesenchymal stem cel transplantation on systemic lupus erythematosus in mice.
METHODS:Sixteen mice with systemic lupus erythematosus were equivalently randomized into control and experimental groups, or then subjected to passage 3 bone marrow mesenchymal stem cel transplantation or the equal volume of normal saline via the tail vein, respectively. Mouse urine samples were colected to detect urine protein levels by Bradford method. Blood samples from the tip of the mouse tail were extracted to detect serum anti-ds-DNS antibody concentration by radioimmunoassay. Mouse kidney tissues were taken and observed pathohistologicaly through hematoxylin-eosin staining and immunohistochemistry staining under microscope. Flow cytometry was used to detect the expression of CD4+CD25+T cels in the inner canthus blood, fresh spleen and thymus.
RESULTS AND CONCLUSION:Within 10 weeks after cel transplantation, the urine protein levels in the two groups were gradualy increased, and the rising velocity was higher in the control group than in the experimental group. From the 4th to 10th week, the urine protein levels in the experimental group were significantly lower than those in the control group (P < 0.05). In the control group, lymphocyte infiltration was visible in the kidney tissues with a few of plasmocytes, and pathological findings showed the mice presented with interstitial nephritis; in the experimental group, the mice had no pathological changes in the kidney. In the two groups, immune complexes were found in the mesangial area, which showed a patch-like distribution in the control group and a punctate distribution in the experimental group; the relative proportion of the occupied area in the experimental group was significantly lower than that in the control group. The expression level of CD4+CD25+T cels in the blood and thymus were significantly higher in the experimental group than the control group (P < 0.05), and the expression level of CD4+CD25+T cels in the spleen was slightly higher in the experimental group than the control group with no significant difference (P > 0.05). The serum anti-ds-DNA antibody concentration in the experimental group was significantly lower than that in the control group (P < 0.05). Taken together, bone marrow mesenchymal stem cel transplantation can improve the pathological damage in systemic lupus erythematosus mice, and has a certain therapeutic effect on systemic lupus erythematosus.

The effect of targeted magnetic nanoparticles on hepatoma and the underlying mechanism were examined. Nude mice transplanted with a human hepatoma cell line (HepG2 cells) were randomized into 5 groups, including: (1) group A, receiving normal saline, (2) group B, receiving 5-fluorouracil (5-Fu), (3) group C, receiving magnetic nanoparticles containing 5-Fu, (4) group D, consisting of treatment with magnetic nanoparticles containing 5-Fu and inside magnetic field and (5) group E, receiving pure magnetic nanoparticles and inside magnetic field. Morphological features of transplanted tumors in mice in each group were observed under transmission electron microscope (TEM). The expression of bcl-2/bax protein was immunohistochemically detected by SABC method. The results showed that a large number of apoptotic tumor cells were found in group B and group D under TEM. The expression of bcl-2 protein was significantly decreased and the expression of bax protein increased significantly in both group B and D as compared with those in group A, C and E (P<0.01 for all). The decrease in bcl-2 and the increase in bax were more in group D as compared with group B (P<0.01). It is concluded that the targeted magnetic nanoparticles containing 5-Fu can improve the chemotherapeutic effect of 5-Fu by decreasing bcl-2 expression, increasing bax expression and inducing apoptosis of the liver cancer cells.

The bile was collected from fro patients with biliary infections, with the bacterium isolated to study the sensitivity of each kind of the bacterium to several antibiotics in common use. Except G- bacterium, we also found some kinds of G+ bacterium in infection bile. G- bacterium were not sensitive to Clindamycin, G+ bacterium were sensitive to Ciprofloxacin. Escherichia coli, Xanthomonas maltophilia, Enterobacter cloacae, Pseudomonas aeruginosa were sensitive to Ampicillin. G+ bacterium were not sensitive to Azactam. Enterococcus faecalis, Enterococcus faecium, Enterobacter cloacae were not sensitive to Ceftazidime. Enterococcus faecalis, Staphylococcus coagulase negative, Staphylococcus epidermidis, Pseudomonas aeruginosa were not sensitive to Ceftriaxone Sodium. We didn't found any bacterium resistance Imipenem. The possibility of the existence of G+ bacterium as well as drug resistance should be considered n patients with biliary infections. The value of susceptibility test should be respected to avoid drug abuse of antibiotics.
Anti-Bacterial Agents/*pharmacology ; Anti-Bacterial Agents/therapeutic use ; Cholecystitis/drug therapy ; Cholecystitis/*microbiology ; Drug Resistance, Bacterial ; Enterobacter aerogenes/drug effects ; Enterococcus faecalis/*drug effects ; Escherichia coli Infections/*drug therapy ; Gram-Positive Bacterial Infections/*drug therapy ; Klebsiella Infections/drug therapy ; Klebsiella pneumoniae/drug effects ; Microbial Sensitivity Tests

Objective To study the theraputic effect of cyclophosphamide,glucocorticoid combined with sildenafil on pulmonary hypertension in systemic lupus erythematosus(SLE?PAH) patients,and to analyze the prognosis adverse factors. Methods Forty?six cases patients with SLE?PAH from Tangshan Gongren Hospital from January 2009 to January 2015 were selected as research subjects. All patients were treated with prednisone, cyclophosphamide and sildenafil,and the course of treatment was 24 weeks. The treatment effect was observed, the systemic lupus erythematosus disease activity score(SLEDAI),6 minutes walk distance and heart function before and after the treatment were compared. The effective and uneffective patients'clinical and test information were compared,and the prognosis adverse factors were analyzed. Results After 24 weeks treatment,the total ef?fective rate was 78. 3%(36/46). Pulmonary artery systolic pressure(PASP) dropped from (59±16) mmHg be?fore treatment to (45±15) mmHg after treatment,there was significant statistical difference(t=-4. 539,P<0. 01). SLEDAI dropped from (6. 6±2. 3) points to (2. 4±0. 8) points,,there was significant statistical differ?ence(t =11. 680,P<0. 01). The 6 minutes walk distance rose form (402±96) minutes to (465±97) minutes, there was significant statistical difference( t=-3. 137,P<0. 01) . Cardiac function improved significantly,there was significant statistical difference( P<0. 01) . There were 10 cases uneffective and deterirated patients,with the age of (42±6) years,36 cases effective patients with the age of (33±4) years,the difference was statistically significant( t=-5. 423, P<0. 01 ) . Uneffective and deterirated patients' anti?RNP antibody positive rate was 90. 9%( 9/10 ) , significantly higher than the effective patients ( 52. 8%( 19/36 ) ,χ2 = 4. 552, P<0. 01 ) . Conclusion Cyclophosphamide, glucocorticoid combined with sildenafil has obvious effect on patients with SLE?PAH. PASP,heart function and 6 minutes walk distance are the important indicators of prognosis. Anti?RNP antibody and age are the adverse prognostic factors.

Objective To explore the regulating mechanism of inducible nitric oxide synthase(iNOS) in hepatic injury of obstructive jaundice (OJ) in vivo and in vitro experiments. Methods (1) Rat hepatocytes were isolated by in situ collagenase perfusion and primary culture. Hepatocytes were pretreated with various concentrations of iNOS inhibitor SMT for 20 min. After pretreatment, 50?M GCDC was added for an additional 24hr. Cells were next detected by FCM and TUNEL.(2) Experimental obstructive jaundice (BDL) was induced by double ligation of the bile duct in rats. After BDL for 3d、7d、14d、and 21d, the apoptotic status in liver of all rats were determined with TUNEL, and iNOS protein in liver of OJ was ditermined with immunohistochemistry method. Results (1) SMT decreased GCDC-induced apoptosis in a concentration-dependent manner. (2) The apoptotic rate of liver was related to length of time of OJ. Apoptosis index (AI) was highest from rats with 14d bile duct ligation. The stronger the iNOS expression, the higher was the number of apoptotic cells that was found in OJ. Conclusions iNOS is involved in the regulation and the occurrence and progression of hepatic injury of obstructive jaundice.

Objective To investigate the clinical efficacy of ozone injection combined with radiofrequency ablation (RFA) for the treatment of far lateral lumbar disc herniation (FLLDH). Methods A total of 60 patients with FLLDH, who were admitted to authors’ hospital during the period from March 2013 to March 2014, were randomly and equally divided into the study group (n=30) and the control group (n=30). Simple ozone injection treatment was employed for the patients of the control group, while ozone injection combined with radiofrequency ablation was adopted for the patients of the study group. The same other adjunctive medication was used in both groups. All the patients were followed up at one week as well as at one, 3 and 6 months after the treatment. The clinical effect and the complications of both g roups were recorded. Results Successful operation was obtained in all 60 patients, no perioperative complications occurred, and the technical success rate was 100%. The excellent clinical results evaluated at one week, and one, 3 and 6 months after the treatment in the control group and in the study group were 73.3%, 76.7%, 70%, 60% and 70%, 76.7%, 83.3%, 90% respectively. The 3-month and 6-month clinical results of the study group were significantly better than those of the control group. Conclusion For the treatment of FLLDH, ozone injection combined with radiofrequency ablation is safe and effective. This technique should be recommended in clinical practice.