Objective To clone lexA gene,express and purify the repressor LexA from Pseudomonas aeruginosa(PA),prepare the polyclonal antibody against PC-1 protein in rabbits,detect immunological activity of LexA protein.Methods The genomic DNA was extracted from the PAO1,the gene fragment encoding the mature LexA was amplied by PCR.It was linked the vector pET32a(+)and expressed in the E.coli BL21(DE3).The expressed protein was purified by two steps of Ni2+ chelate affinity chromatography and gel filtration chromatography respectively.The purified LexA protein immune the rabbits by injection and prepare the polyclonal antibody against PC-1 protein.The immunological activity of expressed and purified LexA protein was detected by ELISA,and Western blot.Results The expressed fused protein was found in insoluble form,accounted for 45% of the total bacteria protein.The final purity was 98.97%,which was determined by the HPLC.The expressed and purified LexA protein had satisfactory immunological activity.

ObjectiveTo investigate the serum prostate-specific antigen (tPSA),serum free PSA to total PSA ratio (f/tPSA),prostate volume (PV) and prostate-specific antigen density (PSAD) in early prostate cancer (PCa) diagnosis.MethodsRetrospective analysis was performed on serum PSA values and related test results from 252 cases of BPH patients and 49 patients with PCa.Prostate volume (PV) was measured by transrectal ultrasound (TRUS),and the f/tPSAand PSAD values were calculated.The differences of serum tPSA,f/tPSA,PV,and PSAD between BPH and PCa group were compared,the area under the ROC curve was used to evaluate these four indicators for its diagnostic sensitivity and diagnostic specificity.ResultsThe values of tPSA,PSAD in PCa group were significantly higher than BPH group ( P ＜0.05),while the values of f/tPSA,PV in PCa group were significantly lower than BPH group ( P ＜0.01orP ＜0.05).The ROC area showed that serum tPSA(0.8013),f/tPSA(0.7390),PV(0.5613) had lower diagnosis value than PSAD(0.9214) in early prostate cancer ( PSAD ＞ tPSA ＞ f/tPSA ＞ PV).When the upper limit of normal PSA was set to take 4ng/ml,the sensitivity was 91.49％,diagnostic specificity was 51.05％.When the f/tPSA threshold set to 0.16,the diagnostic sensitivity was 57.78％,diagnostic specificity was 78.72％.When PSAD threshold was set to 0.15,diagnostic sensitivity was 88.24％,diagnostic specificity was 81.52％.ConclusionsPSA,f/tPSA and PSAD are indicators for biopsy or followup in early diagnosis of prostate cancer.In particular,the diagnostic value of PSAD has higher sensitivity and specificity than PSA and f/tPSA in the diagnosis of prostate cancer.

OBJECTIVETo study the efficacy of Xiaoyao Powder (XYP) combined with interferon alpha (IFN-alpha) in treating HBeAg positive chronic hepatitis B (CHB) patients and the effect on their quality of life (QOL).

METHODSTotally 193 patients with HBeAg-positive CHB confirmed by liver biopsy were randomly assigned to 2 groups, Group A (94 cases) and Group B (99 cases). IFN-alpha1b was subcutaneously injected to patients in Group A at the dose of 50 microg, thrice per week. Those in Group B additionally took XYP. The therapeutic course for all was 24 weeks. Clinical efficacy was observed by assessing ALT restoration rate, HBeAg negative rate, HBeAg conversion rate, HBV DNA negative rate, complete response rate, partial response rate, and symptoms integral. The evaluation of QOL was performed by using chronic liver disease questionnaire (CLDQ) score. Adverse reaction occurrence rate was observed in the two groups.

RESULTSBetter effects were obtained in Group A on ALT restoration rate, HBeAg negative rate, HBV DNA negative rate, complete response rate, partial response rate, TCM symptoms integral, the total effective rate of TCM sysmptoms, CLDQ score, and adverse reaction rates, showing statistical difference when compared with Group B (P < 0.05, P < 0.01).

CONCLUSIONXYP could elevate the efficacy of TCM symptoms of HBeAg-positive CHB patients and anti-viral effect, improve their QOL, and reduce adverse reaction of IFN-alpha.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hepatitis B e Antigens ; Hepatitis B, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Quality of Life ; Treatment Outcome

Objective To evaluate of impact of thalidomide on CD4 +CD25 +T lymphocytes in patients with acute myeloid leukemia and its clinical curative effect.Methods 71 cases of patients with AML were randomly divided into the control group and the experiment group, and 31 healthy people were selected to be the normal group.The experiment group and the control group patients were treated with the same chemotherapy, the experiment group was treated with thalidomide.The levels of CD4 +CD25 +T lymphocyte, CD3 +T lymphocyte, CD4 +T lymphocyte, CD8 +T lymphocyte, ratio of CD4 +/CD8 +and NK cells were detected at before treatment, 10 to 14d after treatment, complete remission 6 months follow-up.In normal group, the same index was detected before and after chemotherapy, and 10-14 d.The clinical curative effect of the experiment group and the control group were observed.Results The effective rate of the experiment group was higher than of the control group(P<0.05); The levels of CD4 +CD25 +T lymphocyte in the experiment group and control group were significantly higher than in the normal group(P<0.05), the two groups decreased compared with the control group, the level of CD4 +CD25 +T lymphocyte in the experiment group was significantly decreased(P<0.05).And with thalidomide treatment, the experiment group in the CD3 +T lymphocytes, CD4 +T lymphocytes and ratio of CD4 +/CD8 +, NK cells were significantly higher than the control group ( P<0.05 ) .Conclusion Thalidomide can improve both the immunity cell function and the clinical efficiency in patients with AML.The mechanism is related to reduce the level of CD4 +CD25 +T lymphocyte.

Objective To investigate the effect of long-term antiviral treatment on clinical outcome and liver histology in patients with hepatitis B virus ( HBV)-related compensated cirrhosis .Methods A total of 61 patients with HBV-related compensated cirrhosis receiving antiviral therapy were enrolled from Taizhou Hospital of Zhejiang Province during September 2010 and March 2015, including 26 HBeAg-positive cases and 35 HBeAg-negative cases .Thirty-nine patients were treated with entecavir ( ETV ) and 22 were treated with adefovir dipivoxil ( ADV ) .Biochemical , serological and virological markers were examined every 3 months during treatment, and Child-Turcotte-Pugh (CTP) scores were calculated.All the patients underwent liver biopsy before and 144 weeks after antiviral treatment .Metavir scoring system was used to evaluate the liver histological activity ( A) and fibrosis score ( F) .Wilcoxon rank sum test and paired t-test were used for the evaluation of liver histopathology and liver function before and after treatment , respectively.Results After 144 weeks of antiviral treatment , HBV DNA was reduced and below the lower limit of detection in 58 patients (95.1%), HBeAg disappeared in 14 patients (14/26, 53.8%), and HBeAg seroconversion was observed in 10 patients (10/26, 38.5%); alanine aminotransferase ( ALT), aspartate amino transaminase (AST), total bilirubin (TBil) and CTP score decreased (t=7.489, 8.259, 14.000 and 6.026, all P<0.01), prothrombin time (PT) was shortened (t=9.777, P<0.01), and serum albumin (Alb) increased (t=3.446, P<0.01).Improvements in both liver histologic activity and fibrosis score were observed (Z=5.716 and 6.657, all P<0.01).Liver histological activity decreased from A1 to A0 in 16 cases, from A2 to A0 in 9 cases, from A2 to A1 in 15 cases, from A3 to A0 in 1 case, from A3 to A1 in 5 cases, and from A3 to A2 in 5 cases.Fibrosis score at the baseline was F 4 for all patients, while after treatment, there were 7 patients with F1, 22 with F2, 20 with F3, and F4 remained in rest 12 patients.Conclusion Both clinical and histological improvements can be obtained after long-term antiviral treatment for patients with HBV-related compensated cirrhosis .

Carcinoma, Hepatocellular ; diagnosis ; Humans ; Liver Neoplasms ; diagnosis ; Serologic Tests

Objective:To explore the clinical effect of "Bian Que flying rescue" system for patients with chest pain in pre-hospital care.Methods:160 patients with acute chest pain were randomly divided into observation groups (80 cases) and control group (80 cases). The patients of observation group received the treatment of "Bian Que flying rescue" system and the patients of control group received the routine pre-hospital emergency measures. A series of indicators including the diagnosis time of pre-hospital, the time span between attack and beginning treatment, clinical curative effect, occurrence of complication and hospital stay time were compared and recorded.Results:The diagnosis time of pre-hospital, diagnosis time of intra-hospital and the time between attack and beginning treatment of observation group were significantly shortened, and the differences of these indicators between two groups were significant (t=14.095,t=9.997,t=4.230,P<0.05), respectively. The numbers of the attack of various complication of observation group were significantly reduced, and the differences of cardiac failure, cardiogenic shock, malignant arrhythmia and myocardial infarction between the two groups were statistically significant (x2=10.230,x2=9.141, x2=5.223,x2=5.096,P<0.05).And the prognosis of observation group was significantly enhanced, and the number of die in one week was significantly reduced, and the differences of these indicators between the two groups were significantly(x2=12.172,P<0.05). Besides, the time of hospital stay and the total cost of hospital stay of observation group were, respectively, significantly shortened and decreased, and the left ventricular ejection fraction (LVEF) of observation group was significantly increased. And the differences of all of these indicators between the two groups were statistically significant(t=4.022,t=4.495,t=9.047,P<0.05).Conclusion: "Bian Que fling rescue" system which was applied in the first aid outside of hospital for patients with chest pain can significantly improve the success rate of rescue, and reduce case fatality rate and the occurrences of complications including congestive heart failure and cardiogenic shock, and shorten the time of therapy, and increase economic efficiency of hospital. It has a certain clinical application value.

OBJECTIVETo observe the efficacy of Zaozhu Yinchen Recipe (ZZYCR) on non-alcoholic steatohepatitis (NASH) patients, and to explore its effect on serum free fatty acid (FFA) and tumor necrosis factor alpha (TNF-alpha).

METHODSTotally 120 patients with NASH were randomly assigned to the treatment group (60 cases, treated with ZZYCR, one dose per day) and the control group (60 cases, treated with Silibin Meglumine Tablets, 20 mg each time, thrice per day). The therapeutic course for all was 24 weeks. Serum levels of ALT and AST activities, TC and TG levels were detected before and after treatment. Peritoneal CT was performed in all patients, and CT ratios of liver and spleen calculated. NAFLD activity score (NAS) and degree of hepatic fibrosis were assessed using pathological examinations of liver tissue, and efficacy also evaluated. Serum contents of FFA and TNF-alpha were also detected.

RESULTSCompared with before treatment in the same group, activities of ALT and AST, serum levels of TC, TG, FFA, and TNF-alpha, NAS, scores of symptoms and signs all obviously decreased, degree of hepatic fibrosis was obviously improved in the two groups (P < 0.05, P < 0.01). These changes were more obviously seen in the treatment group (P < 0.05). After 24-week treatment, the total effective rate and total clinical efficacy were 80.00% (48/60 cases) and 85.00% (51/60 cases) in the treatment group, obviously higher than those in the control group [60.00% (36/60 cases) and 73.33% (44/60 cases) respectively], with significant difference (P < 0.05, P < 0.01).

CONCLUSIONZZYCR could improve the clinical efficacy of NASH patients, and its mechanism might be associated with inhibiting serum levels of FFA and TNF-alpha.

Drugs, Chinese Herbal ; therapeutic use ; Fatty Acids, Nonesterified ; blood ; Humans ; Non-alcoholic Fatty Liver Disease ; drug therapy ; Tumor Necrosis Factor-alpha ; blood

AIM: To investigate therapeutic efficiency of amniotic extraction on dry eye in rabbit model induced by topical benzalkonium chloride (BAC).
METHODS: Totally 26 rabbits (26 right eyes) with dry eye model were studied and divided into two groups:group A (control group with PBS eye drops, n = 13) and group B ( amniotic extraction group, n = 13). Another two rabbits were chosen as normal control. The SchirmerⅠ tests ( S Ⅰ t) and corneal fluorescein staining ( FL) were made, and the tear total protein content, amylase activity, lactoferrin, lysozyme contents, goblet cell density were performed in two groups before treatment and 1, 2, 4 and 8 wk after treatment.
RESULTS: There were significant differences in SIT, FL scores, lysozyme activity and goblet cell density among different groups at different time points (P<0. 05). But, there was no significant differences in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups before treatment (P>0. 05). After 8wks' treatment with PBS, the mean differences of the group A showed great changes in SⅠt, lysozyme and goblet cell density compared with those before treatment ( P < 0. 05); but there was no significant differences in FL scores compared with those before treatment (P>0. 05). As for group B, 8wks after treatment, there were statistical changes in SⅠt, FL, lysozyme (P<0. 05); but there was no significant differences in goblet cell density compared with those before treatment ( P > 0. 05). It was evident that statistical differences were observed in S Ⅰ t, FL scores, lysozyme activity and goblet cell density between two groups at each time point (P<0. 05). However, there were no significant differences in total protein, lactoferrin, amylase activity at different time points (P>0. 05). Meanwhile there was no significant differences in total protein, lactoferrin, amylase activity between two groups before treatment ( P > 0. 05 ). But there were significant differences in total protein, lactoferrin, amylase activity between two groups after 4 and 8 wks'treatment (P<0. 05).
CONCLUSION: Amniotic extraction has significant therapeutic effect on the dry eye in rabbit model.

Human Nestin (hNestin) has been found to express in melanoma,and its expression is positively correlated with the advanced stage of melanoma.However,the precise role of hNestin in the development of melanoma has not been fully understood.The present study aimed to explore the role of hNestin in the proliferation and invasion of melanoma cells.The lentivirus vector carrying a short hairpin RNAs (shRNAs) targeting hNestin (hNestin-shRNA-LV) was stably infected into human melanoma cells UACC903,which expressed high levels of hNestin.The effects of hNestin knockdown on the proliferation,apoptosis,migration of melanoma cells and the related signaling pathways were investigated by immunofluorence,Western blotting and reverse transcription polymerase chain reaction (RT-PCR),respectively.The results showed that hNestin was expressed in most melanoma specimens and the melanoma cells studied.Knockdown of hNestin expression significantly inhibited the proliferation of melanoma cells,blocked the formation of cell colony,arrested cell cycle at G1/S stage and suppressed the activation of Akt and GSK3β.hNestin-silent cells also showed a sheet-like appearance with tight cell-cell adhesion,decreased membrane expression of N-cadherin and β-catenin,and attenuated migration.Furthermore,hNestin silence resulted in the inhibition of tumor growth in vivo.Our study indicates that hNestin knockdown suppresses the proliferation of melanoma cells,which might be through affecting Akt-GSK3β-Rb pathway-mediated G1/S arrest,and hNestin silence inhibits the migration by selectively modulating the expression of cell adhesion molecules in the process of epithelial-mesenchymal transition.