Objective To generate cytokine induced killer (CIK) by inducing lymphocytes cells from peripheral blood in vitro , and to study the phenotype and biological activities of CIK. Methods Lymphocytes cells were isolated fresh from peripheral blood of healthy donors by Ficoll Hypaque density centrifugation, and the cells obtained were resuspended in RPMI medium consisting of 10% fetal calf serum at 37℃, 5%CO 2 for 2h. On day 0, the nonadherent cells were activated with IFN ? (1 000U/ml) and the following days were stimulated with CD3mAb (3 75?g/ml) and rhIL 2 (600U/ml). Phenotype of CIK were analysed by FACS. The proliferation of CIK was tested using 3 H Thymidine, and the killing experiment using MTT tests. Results The proliferation peak of CIK was at day 20, and declined at day 30. The percentages of the cytotoxicity of CIK cells for stomach cancer cell line MGC803 and liver cancer cell line Bel7402 was 65 45%?5 68% and 68 37%?3 93% respectively. CIK cells expressed the phenotypes of CD3CD56, CD3, CD54, CD28CD54, CD11a, HLA DR and CD28. Conclusion IFN ?, CD3mAb and rhIL 2 can induce strong proliferation of peripheral lymphocytes to produce CIK cells expressing the phenotype of CD3CD56, CD3, CD54, CD28CD54, CD11a, HLA DR, CD28, which have strong cytotoxicity on tumor cells.

Objective To evaluate the protective effect of genistein on psoralens plus ultraviolet A (PUVA)-induced photoaging in human dermal fibroblasts (HDFs) in vitro.Methods Dermal fibroblasts were isolated from the foreskin of a healthy 5-year-old boy,and subjected to primary culture.After 5-8 passages of subculture,the fibroblasts were collected and used in the following experiment.To determine the optimal concentration of genistein,methyl thiazolyl tetrazolium (MTT) assay was conducted to detect the proliferation of fibroblasts pretreated with 8-methoxypsoralen (8-MOP) and various concentrations (0-20 μg/ml) of genistein for 24 hours followed by UVA irradiation.Then,the fibroblasts were divided into 3 groups:normal control group receiving no treatment,photoaging group incubated with 8-MOP for 24 hours followed by UVA irradiation,and genistein group incubated with both 8-MOP and genistein at the optimal concentration for 24 hours followed by UVA irradiation.After additional culture,invert microscopy was carried out to observe the morphology of fibroblasts,enzyme histochemistry to assess senescent cells by using SA-β-Gal kit,flow cytometry to determine the level of reactive oxygen species (ROS),real-time fluorescence-based quantitative PCR to detect the matrix metalloproteinase-1 (MMP-1) expression.One-way analysis of variance was conducted to assess the differences in these parameters among these groups.Results At 24 hours after UVA irradiation,the percentage of fibroblasts positive for SA-β-galactosidase was (0.67 ± 0.58)％,(96.67 ± 1.53)％ and (51.67 ± 2.08)％ in the normal control group,photoaging group and genistein group respectively,with significant differences among these groups (P ＜ 0.01).The level of ROS in the photoaging group and genistein group was (0.88 ± 0.24) and (0.62 ± 0.02) fold higher than that in the control group(both P ＜ 0.01).Moreover,the MMP-1 expression level in the photoaging group and genistein group was 10 times and 6 times that in the control group,respectively,with significant differences among the 3 groups (P ＜ 0.05).Conclusion In vitro,genistein can protect against PUVA-induced photoaging in human dermal fibroblasts to some extent.

5) who were conscious and had no obvious cardio-respiratory and hepato-renal dysfunction were enrolled in this study. The patients received intravenous PCA with morphine for the first 48 hours. The PCIA morphine solution contained morphine 2-6mg?ml-1 and droperidol 1-2 mg?30 ml-1 . PCIA included a bolus dose of morphine 1-3 mg with a 5 min lockout. No background infusion was given. Morphine PCIA was then combined with transdermal fentanyl. The initial dose of transdermal fentanyl was 25?g ? h-1 in the first two to three days. The dose was then gradually increased in 25 ?g?h-1 increments according to VAS scores and the amount of Ⅳ morphine needed to reduce the persistent pain until transdermal fentanyl alone could provide sufficient relief of persistent pain and Ⅳ morphine was given only for breakthrough pain. Pain intensity (VAS scores) before and after treatment, daily consumption of morphine (mg?d-1) and transdermal fentanyl (?g?h-1), vital signs and side effects were recorded.Results The 13 patients included 6 males and 7 females. Their mean (?SD) age was 54?15 yrs and body weight 53? 8 kg. There was positive correlation between the titrated dose of transdermal fentanyl (??k-1) and the initial need of Ⅳ morphine (mg?day-1). Y= 1.3606 X + 6.5088. Persistent pain intensity and breakthrough pain intensity evaluated by VAS scores were significantly reduced during the treatment ( P

Objective To investigate the diagnosis and treatment outcomes of testieulax endodermal sinus tumor. Methods Twenty-four cases diagnosed with testieular endodermal sinus tumors from November 1996 to April 2007 were retruspeetively reviewed. Eighteen patients presented with stage Ⅰ disease, 4 presented with stage Ⅰ , and 2 presented with stage Ⅲ. Inguinal radical or-chieetomy were performed in all patients. Retroperitoneal lymph node dissection was performed in 8 ea-ses. Results The histological structures were rather complicated in 24 tumors. Twenty-three eases (96%) were found with reticular pattern, 22(92%) with hyaline body, 20(83%) with glandlike structure, 16(67%) with Shiller-Dural body, and 13(54%) with solid formation. The former 3 kinds were regarded as the main diagnostic criteria. Twenty-one eases were followed up for 20 months to 12 years. During the follow-up, 2 patients died of the disease. Condesions Early diagnosis and combi-nation therapy, including radical orehieetomy and chemotherapy, are the keys to improve the curative effect of testieular endodermal sinus tumors. Active surveillance of AFP is critical for monitoring the recurrence and metastasis of this tumor.

Objective To explore the application of the paries anterior vagina U shape muscularis mucosal flap for the repairing of female complicated urethrovaginal fistula. Methods Fifteen complicated urethrovaginal fistulas patients resulted from severe pelvic frac-ture and with history of failing repairing urethrovaginal fistulas were analyzed retrospectively. The mean diameter of maximum urethrovagi-nal fistula was approximately 2.3 cm. The parias anterior vagina U shape mucosal flaps were used to repair the urethrovaginal fistulas, and the paries anterior vagina muscularis mucosae were used to cover the fistulas surface again. Results Fifteen patients were followed up for 6 to 80 months, and they gained normal voiding, with no urine leakage, urinary incontinence and urethra] stricture occurring. Conclusion The paries anterior vagina U shape mucosal flap is an effective technique for repairing complicated urethrovaginal fistula.

AIM: To study the protective effects of HIF-1α gene transfection on hypoxic injury in human HepG2 cells. METHODS: After gene transfection, HepG2 cells were randomly divided into 4 groups: normoxia with Ad-GFP transfected group, normoxia with Ad-HIF-1 transfected group, hypoxia with Ad-GFP transfected group and hypoxia with Ad-HIF-1 transfected group. LDH leaking rate, cell viability, contents of NO and ROS, the iNOS activity were measured. RESULTS: High levels of HIF-1α mRNA and protein were detected in Ad-HIF transfected HepG2 cells. Cell viability was significantly lower in Ad-GFP transfected-hypoxia group than that in Ad-GFP transfected-normoxia group (P<0.05). No marked difference of cell viability was found between Ad-HIF transfected-hypoxia group and Ad-HIF transfected-normoxia group. ROS was significantly higher in Ad-GFP transfected-hypoxia group than that in Ad-GFP transfected-normoxia group (P<0.05), while no marked difference was found either between Ad-HIF transfected-hypoxia group and Ad-HIF transfected-normoxia group or between Ad-HIF transfected-hypoxia group and Ad-GFP transfected-hypoxia group. The content of NO and iNOS activity were significantly higher in Ad-HIF transfected-normoxia group and Ad-GFP transfected-hypoxia group than those in Ad-GFP transfected-normoxia group (P<0.05), no marked difference was found either between Ad-HIF transfected-hypoxia group and Ad-GFP transfected-hypoxia group or between Ad-HIF transfected-hypoxia group and Ad-HIF transfected-normoxia group. CONCLUSION: Higher HIF-1α expression is contributed to protective effects against hypoxic injury in HepG2 cells, the mechanisms of which may be correlated with promoting expression of gene regulated by HIF-1 and restraining over-expression of injure factors.

Objective To study the prevalence rate of nonalcoholic fatty liver disease (NAFLD)and its correlation with the metabolic syndrome among professional crowd in Yichang city ,and analyze the characteristic of prevalence of NAFLD .Methods Physical check‐up and liver ultrasonography were done and fasting blood GLU ,TG ,HDL‐C ,UA ,CRP were measured for sampling survey professional people in Yichang city .We sampled 6 450 people in 15 company ,including 3 284 men ,3 166 women(20 to 70 years old) ,the results were analyzed .Results The NAFLD prevalence rate of Yichang professional crowd was 21 .71% ,28 .68% in male and 14 .47% in female respectively(P<0 .01) .The prevalence rate in male was higher than that of female before 60 years old . NAFLD prevalence rate in women showed a trend of increasing along with the age growth ,the incidence rate come up to 31 .31%when women were over 60 years old .The highest prevalence rate of MS related components in NAFLD group were obesity (69 .98% )、high blood TG level(61 .10% ) .Conclusion Male before 60 years old and female over 60 years old of Yichang profes‐sional crowd belong to NAFLD high‐risk groups ,the group should be focused on as regular monitoring ,prevention and interven‐tion .NAFLD prevalence rate significantly increased in people with MS .The most important factors of suffering from NAFLD are o‐besity ,high blood TG level .

Objective This article aimed to investigate the treatment adherence of patients with hypertension and examine determinants of adherence.Methods A longitudinal design was adopted to test the treatment adherence and correlated factors of patients with hypertension twice from 2009 to 2012.The influencing factors of treatment adherence in patients with hypertension were analyzed.Results A total of 520 patients finished the first investigation,and 331 patients completed the second investigation.The comparison of demographic characteristics between patients who participated the second investigation or not showed no significant difference except for the education degree (x2=9.38).The incidences of symptoms and complications,systolic blood pressure and diastolic blood pressure of hypertensive patients and the laboratory indexes such as serum creatinine (SCr),total cholesterol (TC),low density lipoprotein (LDL),high density lipoprotein (HDL),fasting blood-glucose and postprandial blood sugar in the second investigation were lower than those of the first investigation,but the difference showed no statistical significance.The scores of treatment adherence,self-efficacy,social support,quality of life were higher in the second investigation than those of the first investigation.The multiple regression analysis found that treatment adherence,social support,education degree and duration of disease course were significant predictors in the first investigation entered the equation,accounting for 26％ of the total variance,among which treatment adherence explained 15％ of the variance,social support 7％,education degree 3％ and duration of disease course 1％.Conclusions The treatment adherence of hypertensive patients improved over one year follow-up.Healthcare providers should pay attention to adherence behavior at initial phase of disease development,and effective strategies targeted patients at risk are suggested to be necessary and should be designed according to the factors affecting adherence.

Objective To investigate and compare the applications of white blood cell count (WBC),neutrophil percentage (N%),C reactive protein (CRP)and procalcitonin (PCT)in the detection of bacterial infections.Methods Patients were randomly recruited in the study,70 patients with bacterial infection disease were recruited in the study as bacterial infection group,81 patients without bacterial infection were enrolled as no infection group.WBC,N%,CRP and PCT were detected,then comparative analysis of test results performed.Results Compared with no bacterial infection group,WBC,N%,CRP and PCT were increased in bacterial infection group(P <0.05),CRP and PCT increased obviously.The positive rate of WBC,N%,CRP and PCT in bacterial infection group was significantly higher than that of no bacterial infection group(P <0.05).In the bacterial infection group,the positive rate of N%,CRP and PCT was significantly higher than that of WBC(P <0.05),the positive rate of CRP was higher than PCT(P <0.05).But the positive rate of CRP was relatively high,and PCT was low in no bacterial infection group,suggesting that the false positive rate of CRP was higher,while that of PCT was lower,which had higher specificity.Conclusion WBC,N%,CRP and PCT all have clinical value for bacterial infections diagnosis.For the diagnosis of bacterial infections,N%,CRP and PCT is superior to WBC.CRP is more sensitive,but less specific,therefore,PCT with higher specificity was more suitable in the diagnosis of bacterial infections.

Objective To investigate the effects of all trans-retinoic acid (ATRA) on the expressions of iron metabolism-related genes and their products in K562 cells and the possible relationship. Methods (1) The characteristics of K562 leukemic cell differentiation induced by ATRA was evaluated by Benzidine, Wright's, NSE and NBT staining.(2) The expression levels of cellular surface antigens (CD71 and CD 13) in K562 cells cultured with ATRA were measured by flow cytometry. (3) IRP/IRE binding activity was assessed by RNA/protein band-shift assay.(4) Ferritin was determined by radioimmunoassay.(5) The mRNA expression levels of H-Fn, TfR and IRP2 in K562 cells cultured with different concentrations of ATRA were delineated by RT-PCR method, confirmed by sequencing of RT-PCR products. Results K562 cells could be induced to differentiate into neutrophils by ATRA, confirmed by cytochemical staining. The expression of CD71 decreased while CD13 increased. The mRNA expression levels of TfR and IRP2 were decreased while mRNA expression level of H-Fn was increased in K562 cells cultured with ATRA, compared to that in control cells. Concomitantly,IRP binding activity was significantly decreased but the level of ferritin was significantly increased in K562 cells cultured with ATRA. Conclusions During the course of K562 cells induction and differentiation to myelocytes by ATRA, the expression level of iron metabolism-related genes and products were changed but the upstream-regulation mechanism still remains unclear.