Objective To investigate the cytotoxic effects of IL-2 combined with different dosages of sorafenib on renal cellular carcinoma cell line 786-0. Methods Renal carcinoma cell 786-0 was cultured. Then , IL-2 (20 μmol/L) combined with different dosages of sorafenib (6.9, 13.8, 20.8 μmol/L) were used to treat tumor cell 786-0. The inhibitory effect on cell proliferation was determined by MTT assay. Cell apoptosis was measured by Annexin V-FITC kit. The tumor-bearing mice models were established and divided into four groups. Results The tumor cell growth was inhibited with the time-course correlation in all groups. In the 48-hour high doses group, the inhibitory rate was up to (74.67±1.87) %. The rates of cell proliferation inhibition and cell apoptosis were higher in the high dosages group than those in the other groups. Conclusions Immunotherapy combined with target therapy could significantly inhibit the growth of renal cellular carcinoma. But we should find a proper dosage, which could improve the clinical effect and reduce the adverse effect.

Objective To study the efficacy of comprehensive treatment for type ⅢA prostatitis.Methods One hundred and eighty-four patients with type Ⅲ A prostatitis, recruited to this study, were comprehensively treated for 8 - 12 weeks by oral antibiotics and α-1 receptor antagonist,indometacin suppository applied into rectal, prostate massage and psychological counseling. The clinical effects of the treatment were evaluated according to the NIH chronic prostatitis symptom index (NIH-CPSI) and leukocyte counts in the expressed prostatic secretions ( EPS ). Results Before and after the treatment, the NIH-CPSI scores were 28. 6 ± 6. 5 and 12. 9 ± 3. 8 ( t = 28. 3, P ＜ 0. 05 ); the pain or discomfort scores were 14. 1 ± 3. 3 and 6. 4 ± 2.2( t = 26. 3, P ＜ 0. 05 ), the urinary symptoms scores were 5.6 ± 1.8 and 2. 1 ± 0. 9 ( t = 23.6, P ＜ 0. 05 ), the scores of life quality were 8.9 ± 3. 1 and 4. 4 ± 2.4 ( t = 15.6, P ＜ 0. 05 ), the leukocyte counts were ( 24. 5 ±4. 4)/HP and ( 6. 2 ± 2. 7 )/HP ( t = 48.1, P ＜ 0. 05 ) respectively, all comparisons showed significantly differences. Seventy-nine cases recovered completely, 57 cases recovered excellently, 36 cases recovered effectively and 12 cases did not recover, the overall effective rate was 93.5%. Conclusion Comprehensive treatment is an effective method for type Ⅲ A prostatitis.

Objective To study the role and clinical significance of chemokine receptor-4 (CXCR4) and vascular endothelial growth factor (VEGF) in the occurrence and development of renal cell carcinoma. Methods Expression of CXCR4 and VEGF were detected by SP immunohistochemical technique in 56 cases of kidney carcinoma tissues (including 20 cases of lymph node metastasis), 10 normal tissues nearby kidney cancer. Results The positive rates of CXCR4 and VEGF were 66. 1% (37/56) and 73. 2% (41/56),which were significantly higher than those in normal tissues( 20. 0% (2/10) and 30. 0% (3/10), respectively) (P ＜ 0. 05 ＝. The expression of CXCR4 protein was significantly positively correlated with that of VEGF protein (r = 0. 315 ,P ＜ 0.05 ＝ in renal cell carcinoma. The expression of CXCR4 and VEGF was closely related to stages of tumor ( χ2 = 9. 520, P = 0. 023; χ2 = 9. 072, P = 0. 027 ), lymphatic metastasis, degree of invasion ( χ2 =4. 972, P = 0. 026; χ2 = 3.910, P = 0. 034 ), and microvessel density ( MVD) ( P ＜ 0. 05 ＝. However, they were not related to sex ( χ2 = 0. 020, P= 0. 887; χ2 = 0. 001, P = 0. 716 ), tumor size ( χ2 = 0. 003, P = 0. 995; χ2 =0. 108, P = 0. 990) and pathologic types ( χ2 = 1. 960, P = 0. 900; χ2 = 0. 112, P = 0. 994). Conclusion There is a significant positive correlation between high expressions of CXCR4 and VEGF proteins in renal cell carcinoma,the high expressions of CXCR4 and VEGF proteins may be related to the metastasis and prognosis of renal cell carcinoma,thus they could be used as important indicators in judging the metastasis prognosis of renal cell carcinoma,and offer prospects for the treatment of renal cell carcinona.

Objective To evaluate the feasibility and the clinical value of extraperitoneal laparoscopic radical prostatectomy in treatment of localized prostate cancer.Methods Clinical data of 26 patients with localized prostate cancer treated with extraperitoneal laparoscopic radical prostatectomy were analyzed retrospectively.All patients were pathologic diagnosed with prostate cancer by preoperative prostate biopsy or transurethral resection of prostate surgery.Gleason grade was from 6-8.Results Twenty-six operations were successfully accomplished ,without converting to open approach.The operative time was 120-270 min(mean was 165 min) ,the intraoperative blood loss was 180-650 ml (mean was 320 ml) ,indwelling catheter time 12-19 d (mean was 14 d).There were 6 cases with little uroclepsia, satisfactory with urination after contract urethral sphincter for 1-3 Months.Pathologically confirmed all prostate cancer;2 cases of positive margins after surgery plus endocrine therapy.All the cases were followed up from 2 to 36 months.The biochemical recurrence was 5 cases who had undergone endocrine therapy.Conclusion Extraperitoneal laparoscopic radical prostatectomy is a safe and feasible procedure with little trauma, small bleeding and fast recovery which is well worth popularizing.Replace open surgery may become frist choice therapeutic method for localized prostate cancer.

OBJECTIVE:To investigate the therapeutic efficacy of edaravone combined with citicoline sodium on acute cere-bral infarction and its effects on the levels of oxidative stress and inflammatory factors. METHODS:108 patients with acute cere-bral infarction were randomly divided into edaravone group(single group)and edaravone+citicoline sodium group(drug combina-tion group),with 54 cases in each group. Based on routine treatment,single group was given Edaravone injection 30 mg added in-to 100 ml 0.9% Sodium chloride injection intravenously,bid,used up within 30 min each time;drug combination group was addi-tionally given Citicoline sodium injection 0.5 g added into 250 ml 0.9% Sodium chloride injection intravenously,qd,on the basis of single group. Treatment course of 2 groups lasted for 2 weeks. NIHSS,HDS,Barthel index,oxidant stress indicator and inflam-matory factors were compared between 2 groups before and after treatment. RESULTS:After treatment,the effective rate of NI-HSS in drug combination group was 81.48%,which was significantly higher than single group(53.70%),with statistical signifi-cance (χ2=9.511,P=0.002). HDS score and Barthel index of 2 groups were significantly increased after treatment,especially in drug combination group,with statistical significance(P<0.05). Compared with before treatment,contents of MDA and ET-1 in 2 groups were decreased significantly,while SOD activity and NO content were increased significantly;the inflammatory cytokines IL-6,IL-8,IL-12 and IL-16,TNF-α were all decreased gradually,with statistical significance (P<0.05);the improvement of each indicator in drug combination group was more significant than single group,with statistical significance(P<0.05). CONCLU-SIONS:Edaravone combined with citicoline sodium show good therapeutic efficacy in the treatment of acute cerebral infarction, can decrease the levels of oxidative stress and inflammation and promote the recovery of the neurological function and the daily liv-ing ability.

Objective To study the relationship between the methylation status of CPG islands in MEG3 gene promoter region of epithelial ovarian cancer and its clinical and pathological features. Methods The promoter methylation status was evaluated by MSP (methylation-specific polymerase chain reaction ) in 47 cases of ovarian cancer tissue and 15 cases of normal control. Results The methylation ratio (42.6%) of the MEG3 genes in the ovarian cancer was statistically significantly higher (P = 0.035 ) than that (13.3%) in the normal control. The methyation rate of the group with an age > 60 years old was slightly higher than that of the group with an age≤60 years old, without statistically significant (P > 0.05), so was observed in ovarian cancers of stage Ⅰ andⅡ than that in stage Ⅲ and Ⅳ. There were also no significant differences in MEG3 gene methylation positive rate neither in different pathological grading nor in various ovarian cancer tissues (P > 0.05). Conclusion Abnormal methylation in MEG3 gene may be associated with epithelial ovarian cancer , but no relation to its clinical pathology.

Objective To evaluate the expression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT-1) in ovarian cancer cell lines, and to investigate the biological effects of down-regulated MALAT-1 on OVCAR3 cells.Methods qRT-PCR analysis was used to examine the expression level of MALAT-1 gene in ovarian cancer cells, including ES-2, A2780, SKOV3 and OVCAR3 cell lines.For functional research, four shRNA oligos specially targeting MALAT-1 and a empty vector were designed and constructed into pGPU6/GFP/Neo, then transfected into OVCAR3 cells.qRT-PCR was used to confirm the effective suppression of MALAT-1.Changes of proliferation and adhesion of cells were analyzed by CCK-8 and adhesion assays.Wound-healing, transwell migration and invasion assays were used to examine migration and invasion of MALAT-l-silencing cells in vitro.Results The expression of MALAT-1 gene in OVCAR3 cells was high, and qRT-PCR results confirmed successfully the knockdown of MALAT-1 after transient transfection.After successful suppression of MALAT-1, the proliferation, wound-healing and adhesion ability in vitro were inhibited to some degree.In transwell migration assay, the number of migration cells in MALAT-1-silencing group was 52.17±4.48, which is much less than that in the negative and control groups (286.50± 12.23 and 295.67±6.96, respectively).In invasion assay, the number of invasion cells passing the transwell membrane in MALAT-1-silencing group (37.33±2.40) was also decreased significantly, compared to that in the negative and control groups (239.00±15.72 and 222.67±20.85, P ＜ 0.05).Conclusions shRNA-mediated silence of MALAT-1 can effectively inhibit the proliferation, adhesion, migration and invasion abilities of ovarian cancer cell line OVCAR3 in vitro, indicating MALAT-1 is expected to be a target gene for the treatment of ovarian cancer.

OBJECTIVETo investigate the reversal effects of different concentrations of DNA methylation inhibitor, 5-aza-2-deoxycytidine, on the hypermethylation of maternally expressed gene 3 (MEG3) gene promoter, and then the inhibitory effect of restoration of MEG3 expression on the proliferation of ovarian cancer cells.

METHODSHuman ovarian cancer OVCAR3 cells were treated with various concentration of 5-aza-2-deoxycytidine (0, 1, 5, 10, 20 µmol/L, respectively) for 6 days. Then the methylation status of MEG3 promoter was detected by methylation specific PCR (MSP). The alteration of MEG3 gene expression was detected by RT-PCR. Cell proliferation was determined by MTT assay and EdU incorporation assay.

RESULTSAfter treated with 5-aza-2-deoxycytidine, the methylation status of MEG3 in the 0, 1, 5, 10, 20 µmol/L 5-aza-2-deoxycytidine groups were 1.00 ± 0.00, 0.79 ± 0.00, 0.67 ± 0.00, 0.65 ± 0.03 and 0.61 ± 0.01 folds, respectively (P < 0.05 for all). The relative expressions of MEG3 mRNA in the 0, 1, 5, 10, 20 µmol/L 5-aza-2-deoxycytidine groups were 1.00 ± 0.00, 2.04 ± 0.16, 2.44 ± 0.17, 3.19 ± 0.34 and 5.34 ± 0.39, respectively (P < 0.05 for all). In contrast to the negative control, the inhibition rates of the OVCAR3 cell growth were increased significantly when treated with 1, 5, 10, 20 µmol/L 5-aza-2-deoxycytidine in 2, 4 and 6 days. There were (40.78 ± 0.80)%, (35.65 ± 0.33)%, (31.81 ± 0.66)%, (27.33 ± 1.27)% and (17.75 ± 1.85)% of EdU-positive cells in the 0, 1, 5, 10 and 20 µmol/L 5-aza-2-deoxycytidine groups (P < 0.01 for all).

CONCLUSIONSMaternally expressed gene 3 promoter hypermethylation is reversed by 5-aza-2-deoxycytidine in ovarian cancer cells. The downregulation of MEG3 gene might be resulted from the methylation, and the re-expression of MEG3 partly contribute to the growth inhibition of epithelial ovarian cancer cells.

Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; DNA Methylation ; Female ; Humans ; Neoplasms, Glandular and Epithelial ; Ovarian Neoplasms ; Promoter Regions, Genetic ; RNA, Messenger