Objective To investigate expressions of programmed cell death ligand 1 (PD-L1) in hepatic tissues at the different stages of hepatitis B virus ( HBV) infection, and clarify its role in the mechanism of chronic hepatitis B virus infection. Methods The expressions of PD-L1 were detected by immunohistochemistry and computer image quantitative analysis in the hepatic tissues of 65 chronic HBV infected patients and 5 healthy controls. The correlations between PD-L1 expression and inflammatory grading in the hepatic tissues, total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum HBV DNA level were analyzed. Results The PD-L1 expressions in hepatic tissues of HBV infection with G0 - G4 inflammatory grades were 3. 07 % ±0.93%, 8.01%±1.49%, 11.60%±2.60%, 18.41%±2.21% and 26. 04% ±3. 41%, respectively,which were all significantly stronger than that in controls (0. 64%±0. 28%). PD-L1 expression was a positively correlated with inflammation grading of hepatitis tissues, TBil, ALT and AST level in serum (r=0. 917, 0. 787, 0. 483, 0. 628; all P<0. 05), and negatively correlated with serum HBV DNA load (r=-0. 620, P<0. 05). Conclusion The upregulated PD-L1 expression may be probably involved in the chronicity of HBV infection.

ObjectiveTo observe the short-term efficacy of treatment with the diclofenac diethylamine emulge import by pulsed ultrasound, massage therapy and quadriceps muscle strengthen training to the patients with knee osteoarthritis for elderly.MethodsOn the basis of health education, 30 elderly patients with knee osteoarthritis were accepted treatments with 10 times of diclofenac diethylamine emulge import by pulsed ultrasound, 6 times of massage therapy and then they completed quadriceps muscle strengthen training once or twice a day.ResultsThe pain, walking ability, sit-to-stand ability and lower extremities muscle strength improved significantly for the patients after treatments (P<0.05 or P<0.001). ConclusionThe diclofenac diethylamine emulge import by pulsed ultrasound, massage therapy and quadriceps muscle strengthen training have a good short-term efficacy to the elderly patients with knee osteoarthritis.

Objective:To determine the basic characteristics and clinical manifestations of hospitalized patients with psoriasis in Hunan, and to provide reasonable reference for the etiology and treatment of psoriasis.
Methods:Totally 190 patients with psoriasis from January 1, 2012 to December 31, 2012 treated in the Department of Dermatology, the Second Affiliated Hospital of Central South University were retrospectively analyzed. The data were analyzed by SPSS17.0.
Results:The ratio of male to female inpatient numbers was 3.13:1, aged was between 40 and 70 years. The most common subtype of psoriasis was psoriasis vulgaris (64.73%), followed by psoriatic erythroderma (18.42%). The distribution of the subtype of psoriasis did not differ by gender. Nineteen patients recalled clearly the cause of proriasis, 5 of which were caused by medicine, and 4 by drinking. Totally 96 cases accompanied with other diseases, 24.21%of which accompanied with cardiovascular system disease.
Conclusion:There is no season difference in the hospitalization of patients with psoriasis. there are more male than female inpatients. Treatment of psoriasis should consider clinical classification, drug adverse reactions, and individual factors for individual treatment.

Objective To study the dynamic changes of Hepcidin-25 in different disease process in patients with multiple myeloma (MM) and its relationship with therapeutic effect and prognosis.Methods The expression levels of Hepcidin-25 in 54 MM patients were analyzed by RT-PCR and ELISA.The correlations between dynamic changes of patients' Hepcidin-25 expression and clinical manifestations,clinical predictive value of the prognosis were researched.Results Expressions of Hepcidin-25 mRNA and protein in MM patients were significantly higher than those in normal control group [0.58±0.19 vs 0.08±0.027,P =0.001,(5.2±11.9) ng/ml vs (22.5±7.1) ng/ml,P =0.019].Hepcidin-25 expression of 17 patients remission after treatment was (26.4±7.3) ng/ml,it was significantly lower than that before treatment [(33.4±7.4) ng/ml,t =5.312,P =0.021].But the Hepcidin-25 level of the treatment ineffective patients (37 cases) significantly increased [(55.9±12.7) ng/ml vs (39.1±9.9) ng/ml,t =2.811,P =0.037].There existed obvious differences in survival curves of two groups (P ＜ 0.01).Hb level of patients remission after treatment was 124 g/L,and significantly higher than that before treatment (102 g/L,t =2.113,P =0.035).It had a negative correlation with Hepcidin-25 (r =-0.535,P =0.002).But the Hepcidin-25 level of the treatment ineffective patients significantly increased (46 g/L vs 73 g/L,t =2.730,P =0.036).It also had a negative correlation with Hepcidin-25 (r =-0.642,P =0.001).Conclusions Hepcidin-25 expression levels in patients with MM are high,and are closely related to anemia of chronic disease.High expression of Hepcidin-25 has a predictive value for clinical effect and prognosis.

OBJECTIVETo demonstrate the effects and mechanisms of Qifu decoction( QFD) on renal interstitial fibrosis (RIF) in model rats with yang-deficiency syndrome.

METHODThe rats were randomly divided into 3 groups, the Sham group (Group A), the Model group (Group B), the Qifu decoction group (Group C) and the Enalapril group (Group D). The RIF model was established by adenine administrated and unilateral ureteral obstruction (UUO) of the left ureter. After the model was successfully established, the rats in Group C and D were administrated with QFD or the Enalapril suspension,while the rats in Group A and B were administrated with distilled water. All rats were administrated for 3 weeks. Before administration and at the end of week 1, 2 and 3, the rats were weighted, and 24 h urinary protein excretion (Upro), urinary β2-microglobulin (Uβ2-MG) and urinary N-acetyl-D-glucosaminidase (NAG) were examined, respectively. All rats were killed after administration for 3 weeks. Blood and renal tissues were collected, renal morphology and tubulointerstitial morphology were evaluated, respectively. Serum cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), blood urea nitrogen (BUN), serum creatinine (Scr) and uric acid (UA) were detected, respectively. The protein expressions of E-cadherin, α-smooth muscle actin(α-SMA), transforming growth factor-β1 (TGF-β1), onnective tissue growth factor (CTGF) extracellular signal-regulated protein kinase 1/2(ERK1/2) and phosphorylated-ERK1/2 (p-ERK1/2) in kidney were evaluated, respectively.

RESULTQFD ameliorated serum cAMP level and the rate of cAMP/cGMP, attenuated urinary β2-MG level, NAG level and renal tubulointerstitial fibrosis, increased E-cadherin protein expression, and reduced α-SMA, TGF-β1, CTGF and p-ERK1/2 protein expressions in the kidney. However, QFD had no influence on renal function in vivo. In addition, these effects were better than those of the model rats treated by Enalapril.

CONCLUSIONQFD could alleviate yang-deficiency parameters, as well as urinary β2-MG level and NAG level in model rats induced by adenine administration and UUO. Moreover, QFD could improve EMT and RIF by up-regulating E-cadherin protein expression, and down-regulating α-SMA, TGF-β1, CTGF and p-ERK1/2 protein expressions, the key molecular in ERK1/2 signaling pathway.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; Fibrosis ; Kidney ; drug effects ; pathology ; Kidney Diseases ; drug therapy ; pathology ; MAP Kinase Signaling System ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Ureteral Obstruction ; complications ; Yang Deficiency ; complications

Objective: To explore the etiology, clinical manifestations, pathological features, diagnosis and treatment of small-cell carcinoma of the endometrium. Methods: This case report introduces a case of small-cell carcinoma of the endometrium and reviews the related literatures. Results: This patient presented with vaginal bleeding and was diagnosed of stage II endometrial adenocarcinoma before surgery. Laparoscopic modified radical hysterectomy and double accessory and pelvic lymph node dissection was performed. Pathological examination revealed small-cell carcinoma of the endometrium complicated with stage II endometrial adenocarcinoma. There was immunohistochemical evidence of synaptophysin expression. Chemotherapy with EP regimen (etoposide plus carboplatin) and radiotherapy were administered after surgery. Conclusion: Primary small-cell carcinoma of the endometrium is an extremely rare and aggressive disease with a poor clinical outcome. Surgery combined with chemotherapy and radiotherapy may contribute to an improved prognosis.

Objective To investigate the effect of neutrophil extracellular traps (NETs) on inflammation of rheumatoid arthritis (RA), especially angiogenesis. Methods The presence of NETs in synovial tissues of RA and osteoarthritis (OA) was observed by immunofluorescence assay. Neutrophils were isolated from peripheral blood of health volunteers. Neutrophils were cultured in vitro, the formation of NETs was observed. NETs were extracted as a stimulating agent. The effects of NETs on the proliferation of human umbilical vein endothelial cells (HUVECs) and synovial fibroblasts (RAFLS) were evaluated by MTT, and which were classified into two groups: HUVECs group and RAFLS group, with the following treatment: control and NETs (0.28 mg/L). Wound repair assay was employed to evaluate the effect on the cell migration stimulated with NETs. The experiment was divided into three groups:control, VEGF (40μg/L VEGF) and NETs (0.28 mg/L NETs). Results (1) Compared with OA, NETs were found more in the synovial tissue of RA. (2) NETs formation was induced by stimulator in vitro. The concentration of extracted NETs-DNA was 27.8 mg/L. (3) MTT assay showed that compared with the control groups, low concentration of NETs (0.28 mg/L) promoted the proliferation of HUVECs (0.499 ± 0.011 vs. 0.393 ± 0.009, P<0.05) and RAFLS (0.266 ± 0.007 vs. 0.192 ± 0.007, P<0.05). (4) It was showed that a significant wound closure induced by low concentration of NETs (0.28 mg/L) was found compared with control. Conclusion Our present study suggests that NETs are found more in the synovial tissue of RA, and low concentration of NETs can promote angiogenesis in RA.

Objective To explore whether serum amyloid A (SAA) can induce the formation of neutrophil extracellular traps(NETs)in neutrophils in vitro. Methods A stable method for inducing NETs formation in vitro was established, in-cluding isolation of peripheral blood neutrophils, cell culture, and NETs formation and observation. The neutrophils were iso-lated from peripheral blood of healthy volunteers. And cells were cultured in vitro and classified into three groups:negative control (NC) group, SAA group and lipopolysaccharide (LPS) group. Following the distinct stimulation in three groups, NETs formation was observed and its percentage was calculated. The concentration of hinstone (h) 3 in supernatant was detected by ELISA. Results The purification and vitality of isolated neutrophils were both more than 95%. The nuclei of neutrophils lost their shape and spread, NETs formation was found. More NETs formation was found in SAA group than that in NC group (P < 0.05). Moreover, the concentration of h3 in supernatant was significantly higher in SAA group than that in NC group (P<0.05). Conclusion SAA can induce the formation of NETs in vitro.

Objective To investigate the effects of different doses of alcohol on the synthesis of testosterone and the expression of androgen binding protein(ABP)mRNA in rat testis.Methods Forty male Wistar rats were randomly divided into 4 groups(10 rats each group)and received either distilled water(control group)or alcohol(alcohol-fed groups)for 5 months.Alcohol was administered by garage with a single daily dose : 5 g/kg(large dose group),2.5 g/kg(middle dose group)and 0.5 g/kg(small dose group).Testosterone content was measured by ELISA.mRNA levels of peripheral-type benzodiazepine receptors(PBR),PPARct and ABP were assayed by RT-PCR.Results Compared with control group:(1)ethanol feeding with daily doses of 5 g/kg,2.5 g/kg and 0.5 g/kg significantly decreased testosterone levels by 31.13%(P0.05)respectively,indicating that ethanol might impair testosterone synthesis;(2) mRNA levels of PBR were decreased in all three ethanol-treated groups(all P

Objective To purify Shiga toxin Ⅱ (STX Ⅱ) of enterohaemorrhagic Escherichia coli (EHEC) O157: H7 by affinity chromatography, and characterize its biological function. Methods The immno-affinity chromatography column was prepared by STX Ⅱ A subunit-specific antibody S1D8 coupling to Sepharose 4B matrix. The purity and specificity of STX Ⅱ molecule secreted by EHEC O157:H7 were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, respectively. The purified toxin was serially diluted and the toxic activities to Vero cell line and mice were observed. The 50% cytotoxic dose (CD50) for Vero cell line and 100% lethal dose (LD100) for mice were calculated. The protection effect of anti-STX Ⅱ polysera to the mice against the purified toxin challenge was also observed. Results STX Ⅱ was successfully purified from culture supernatant of EHEC O157:H7 using affinity chromatography scheme. The relative molecular weights of STX Ⅱ A and B subunits were 32 000 and 7 500 confirmed by SDS-PAGE, respectively. The purified toxin could react with monoclonal antibodies against STX Ⅱ A and B subunits, respectively.The toxin was cytotoxic to Vero cell with CD50 of 20 ng/L and lethal to mice with LD100 of 5 ng.The toxin could be neutralized by anti-STX Ⅱ polysera in vivo. Conclusion STX Ⅱ is successfully purified and its toxic effects are confirmed in both cell line and mouse model.