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.

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 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.

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 investigate the correlation between expression of human telomerase reverse transcriptase (hTERT) mRNA in peripheral blood and postoperative recurrence and prognosis in the patients with advanced colorectal cancer (CRC).Methods The preoperative and 1 month postoperative quantitative expression of hTERT mRNA in peripheral blood in 85 patients with advanced CRC were detected with real-time fluorescence quantitative real-time fluorescence quantitative polymerase chain reaction (RTPCR).The quantitative expression of hTERT mRNA in peripheral blood in 30 normal controls were also detected.The postoperative prognosis was followed-up,and the quantitative expression of hTERT mRNA in peripheral blood in the patients with recurred CRC and the patients without recurred CRC were detected repeatedly.The factors that affected the preoperative hTERT mRNA expression were analyzed.The relationship was investigated between the hTERT mRNA expression levels and postoperative recurrence and prognosis in the patients with advanced CRC.Results The hTERT mRNA expression levels in peripheral blood in advanced CRC group were significantly higher than those in the normal controls [(6.22 ±5.85)2-△△Ct vs (0.45 ±0.27)2-△△Ct,(P ＜0.01)],and were significantly decreased at the time point of 1 month after the operation (P ＜ 0.01).The hTERT mRNA expression levels in the postoperative recurred CRC group were significantly higher than those in the 1 month postoperation group and no recurrence CRC group [(5.07 ± 2.87)2-△△Ct vs (1.83 ± 1.08)2-△△Ct,(2.15 ± 1.49)2-△△Ct,(P ＜0.01)].Univariate analysis revealed that preoperative high hTERT mRNA expression levels in peripheral blood were related to the differentiated degree of tumor,tumor size,lymph node metastasis ratio,and liver metastasis (P ＜ 0.05 or P ＜ 0.01).Multivariate logistic regression analysis revealed that differentiated degree of the tumor,lymph node metastasis,and liver metastasis were independent risk factors for the elevation of preoperative hTERT mRNA expression in the advanced CRC.Elevated preoperative hTERT mRNA expression was positively related to the postoperative recurrence rate,and was negatively related to postoperative survival rate.Conclusions Preoperative hTERT mRNA expression levels in peripheral blood were related to the important pathologic features in patients with advanced CRC,and might be a valuable prognostic factor.Postoperatively elevated hTERT mRNA expression in peripheral blood was related to recurrence of advanced CRC.

AIM: To investigate the effect of antisense oligonucleotides(ASON) of c-sis on cellular cycle and proliferation of pulmonary artery vascular smooth muscle cells(VSMC).METHODS: Tissue mass culture was done to get VSMC of pulmonary artery. Different concentrations of antisense oligonucleotides of c-sis were added into the cultures to observe the VSMC proliferation curve using MTT test. The changes of VSMC cellular cycle were also observed by flow cytometry. RESULTS: ASON with mid-to high concentrations restrained the proliferation of VSMC apparently with the peak of cell growth being attenuated or eliminated. Affected by mid-concentration ASON, PDGF-BB showed significant accelerating effect on the proliferation of VSMC. The ratio of G 0/G 1 in cellular cycle was increased significantly in VSMC culture with ASON in comparison with control. The G 0/G 1 ratio also showed significant differences among different concentration of ASON groups( P

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 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.

This study aimed to optimize the ultrasonic extraction of gardenia oils from Gardenia jasminoides Ellis.by response surface analysis methodology (RSM).The duration of extraction,ratio of liquid to materials and extracting times were selected as impact factors based on single-factor experiment.RSM was adopted to learn the effects of the three factors on the extraction rate of Gardenia oil from Gardeniajasminoides Ellis.As a result,it was found that the duration of extraction was 28.7 min,while the ratio of liquid to materials was 11.62:1,and the number of extracting times were 3.36.Under this extracting condition,the average extraction rate of the oil was 13.5％ according to three validation experiments,fitting the predicted value well.In conclusion,it was demonstrated that the ultrasonic extraction method effectively improved the extraction rate of gardenia oil,providing a certain basis for the development and utilization of gardenia oil.

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 .