Objective To investigate the effects of nuclear factor-kappaB(NF-κB)/p65 signaling transduction on the apoptosis and expressions of apoptosis-related genes Bax in nude mouse lung tumour cell xenografts. Methods The nude mice Lewis lung carcinoma cell xenograft model was established,and then the mice were intraperitoneally injected with NF-κB/p65 small interfering RNA(siRNA). The apoptosis of xenografted tumor cells in nude mice was detected by TUNEL assay. Expressions of Bax mRNA and protein were detected by RT-PCR and Western blotting. Results The result of TUNEL assay demonstrated that p65 siRNA evidently evoked cell apoptosis. Compared to the PBS treatment group or the normal control mice,both mRNA and protein expression levels of Bax in tumor xenografts were significantly up-regulated. There were significant differences among three groups(P<0.05). Conclusion NF-κB/p65 subunit may play an essential role in cell apoptosis of Lewis lung tumor.

Objective To compare the efficacy of the therapeutic alliance of radiotherapy and Semustine (CCNU)chemotherapy with radiotherapy alone after partial resection of high-grade sliomas.Methods Forty-nine cases with partial resection of brain gliomas were randomly divided into two groups.In the control group(sole radiotherapy n=19),patients were only treated with local radiotherapy,with the dose of 40Gy/20f/4W,and then added the amount to 61～64Gy/27～28f/5～6W with low fractionation radiotherapy.In the observation group(therapeutic alliance n=23),patients underwent the above mentioned radiotherapy plus six Courses of adjuvant CCNU chemotherapy.Meanwhile,enhanced GD-MRI was performed to evaluate the changes of glioma volume,the nerve function status was evaluated with Karnofsky scale,and record the survival time.Results Twenty-four weeks after radiotherapy,improvements in glioma size and nerve function status of the observation group were superior than that in the control group(all P＜0.05),meanwhile,the long-term survival rate of observation group war also significantly better than that of the control group(P＜0.05).Conclusion Radiotherapy combined with CCNU could increase the glioma remission rate after partial resection,improve the life quality of patients.and extend their survival time.

Objective:To evaluate the efficacy of silymarin on type 2 diabetic nephropathy in rats. Methods:Totally 60 male al-bino Wistar rats at the age of 8 weeks were involved in the study, and non-insulin-dependent diabetes mellitus was induced in 45 over-night-fasted rats by intramuscular injection of streptozotocin and nicotinamide. Among the 45 rats, three ones failed to make the model, and forty-two rats with diabetic nephropathy were randomly divided into diabetes control group (DC group), diabetes treated by low dosage silymarin group ( DT-L group) and diabetes treated by high dosage silymarin group ( DT-H group) with 14 cases in each. The dosage of silymarin in DT-L group and DT-H group was 60 mg·kg-1 ·d-1 and 120mg·kg-1 ·d-1 , respectively, and the treatment course was 60 days. The other 15 rats were divided into the normal control group ( NC group) with equal volume buffer solution. The glycosylated hemoglobin, urine volume, serum creatine, serum uric acid, and urine albumin levels were analyzed by the semi-automat-ic biochemial analyzer. Results:At the end of the study, the DC group had significantly higher levels of blood glucose, glycosylated hemoglobin, urine volume, serum uric acid and urine albumin than the NC group (P<0. 05), while the two silymarin-treated groups showed significantly lower levels of the indices than the DC group (P<0. 05). Conclusion:The results show that silymarin has pro-tective effect on kidneys of rats with type 2 diabetes mellitus, which lay foundation for the future clinical application.

To study the chemical constituents of Trichosanthes kirilowii Maxim., chromatographic methods such as D101 macroporous resin, silica gel column chromatographic technology, Sephadex LH-20, octadecylsilyl (ODS) column chromatographic technique and preparative HPLC were used and nine compounds were isolated from a 95% (v/v) ethanol extract of the plant. By using spectroscopic techniques including 1H NMR, 13C NMR, 1H-1H COSY, HSQC and HMBC, these compounds were identified as 5-ethoxymethyl-1-carboxyl propyl-1H-pyrrole-2-carbaldehyde (1), 5-hydroxymethyl-2-furfural (2), chrysoeriol (3), 4'-hydroxyscutellarin (4), vanillic acid (5), alpha-spinasterol (6), beta-D-glucopyranosyl-a-spinasterol (7), stigmast-7-en-3beta-ol (8), and adenosine (9), separately. Among them, compound 1 is a new compound, and compounds 3, 4 and 5 are isolated from the genus Trichosanthes kirilowii Maxim. for the first time.

As an important member of metabotropic glutamate receptors (mGluR), metabotropic glutamate receptor 1 (mGluR1) plays an important role in the signal transduction of central nervous system. Selective mGluR1 antagonists can block the signaling pathway activated by mGluR1 and exert a series of physiological actions including analgesia, antianxiety, antidepression, etc. Currently, the discovery and modification of selective mGluR1 antagonists have become a hot research focus. This paper reviews the structural catalogs of selective mGluR1 antagonists and their structure-activity relationships in the last decade.

Objective To investigate the expression and clinical significance of Sir2?related enzymes?1(SIRT1)in colorectal cancer. Methods The expression of SIRT1 was evaluated by immunohistochemistry in 86 selected cases of primary Colorectal cancer and 30 samples of normal rectum tissues besided carcinoma. The relationship of the expression and clinical pathological features were analyzed. Results The positive expression rate of SIRT1 in Colorectal cancer tissue was 88.4%,and significantly higher than in normal rectum tissue besided carcinoma(P<0.001). There were no correlation between the expression of SIRT1 and sex,age and tumor diameter,and significant positive correlation between the expression of SIRT1 and TNM stage (P<0.05),lymph node metastasis(P<0.05),infiltrating depth(P<0.05),and negagtive correlation between the expression of SIRT1 and tumor differentiation(P<0.05). Conclusion The positive expression rate of SIRT1 in colorectal cancer tissue was significantly higher than in normal rectum tissue besided carcinoma and intimate correlation with tumor differentiation,TNM stage,lymph node metastasis and Infiltrat?ing depth. Conclusion SIRT1 may be an important assistance gene to diagnose and judge prognosis,and may improve diagnostic rate and auxiliari?ly judge prognosis as a important Colorectal cancer marker.

Objective To establish real-time PCR method for the detection of JAK2 V617F mutation and evaluate its clinical significance in patients with myeloprollferative disorders and leukemia.Methods 71 chronic myelocytic leukemia(CML) patients, 22 essential thrombocythemia (ET) patients, 11 primary myelofibrosis (PMF) patients, 9 polycythemia vera (PV) patients and 7 cosinophilia patients were enrolled in this study. JAK2 V617F mutation was determined by real-time PCR and amplification refractory mutation system (ARMS), followed by sequencing. Human erythroleukemia cell (HEL cell)DNA was used as homozygous control of JAK2 V617F mutation. The detection limit for either real-time PCR or ARMS was evaluated. Results Real-time PCR assay showed that there was a melting temperature(Tin) peak at (75.0±0.2)℃ for wild type samples and a Tm peak at (76.6±0.2)℃ for mutation type samples. JAK2 V617F mutation was detected in 8(88.9%) patients with PV, 12(54.5%) patients with ET and 7(63.6%) patients with PMF respectively. But there was only one positive case in 71 CML patient (1.4%). The results showed complete concordance with ARMS results and confirmed by sequencing. The mutation could be detected in 102 HEL cells per 106 white blood cells by real-time PCR, whereas the mutation can be assessed in 104 HEL cells per 106 white blood cells by ARMS. Thus, the sensitivity of real-time PCR was 100-fold higher than ARMS. Conclusions The real-time PCR method is successfully established for detection of JAK2 V617F mutation. This method is more sensitive, convenient than ARMS, and suitable for clinical application. There is high frequency of JAK2 V617F mutation in myeloproliferative disorders and it could be used as the diagnostic marker for myeloproliferative disorders.

Objective To compare the efficacy and safety of S-1 plus nedaplatin and standard second-line chemotherapy in the treatment of advanced lung adenocarcinoma that failure in the first-line chemotherapy.Methods A total of 95 cases with IIIb-IV stage non-small cell lung adenocarcinoma that failure in the first-line chemotherapy was included in this paper and divided into two groups:group A(n=5 1 )was treated with S-1 combined with nedaplatin chemotherapy;group B(n=44)was treated with pemetrexed or docetaxel single-agent chemotherapy,21 days were a cycle of treatment,and totally 4 cycles of chemotherapy. Results The objective response rates(CRR)in group A and group B were 33.0% and 19.3%respectively(P<0.05),and the difference was significant;the clinical benefit rates(CBR)in two groups were 57. 1% and 40.9% respectively (P <0.05 ),and there was significant difference between two groups. The median perfect forward secrecy(PFS)was 99 days and 40 days respectively,and the difference between two groups was significant(P<0.05 ). The one-year survival rates in groups A and B were 5 1.3% and 33.7%(P<0.05 ),and there was significant difference(P<0.05 ). Logistic multi-factor regression analysis showed that the ORR and CER had no correlation with the gender, age, proportion score (PS )and the first-line chemotherapy selection. Conclusion S-1 plus nedaplatin chemotherapy is better than the standard second-line chemotherapy in the treatment of advanced lung adenocarcinoma that failure in the first-line chemotherapy,with the advantage of good tolerance,and it can provide a new choice for the treatment of advanced non-small cell lung adenocarcinoma.

Objective To observe the expression change of embryonic dry related genes in embryonic stem of pancreatic cancer and analyze its significance.Methods Pancreatic cancer cell lines PANC-1 was incubated with with CD24,CD44 antibody incubation,the CD24 + and CD44 +pancreatic cancer stem cells were sorted and pancreatic cancer stem cells' forming speed were observed under a microscope.The differentiation of stem cells were induced by 1640 culture,and the expression of CD24 +and CD44 +were detected by flow cytometry.The expression of embryonic stem cell marker Oct4 and Nano were detected by immunohistochemical and the tolerance difference of Oct4 and Nano cell to chemotherapeutic drugs in vitro were observed and compared by CCK8 method.Results The number of CD24 +and CD44 +isolated from human pancreatic cancer cell lines PANC-1 was 1%~3%.Oct4 and Nano expression in sorted cells were significantly higher than un -sorted cells,and the difference was significant (P<0.05 ). Conclusion CD24 +and CD44 +cells has high drug resistance and other characteristics of pancreatic cancer stem cells,and the expression of embryonic stem-related genes Oct4 and Nano are high pancreatic cancer stem cells.

Objective To investigate the effect of hydrogen on endotoxin-induced expression of zonula occludens-1 (ZO-1) in human colon epithelial cells (Caco-2 cells).Methods Caco-2 cells were cultured routinely,seeded in Transwell chambers or wells,and randomly divided into 4 groups (n =45 each) using a random number table:control group (group C);hydrogen-rich culture medium group (group H);endotoxin group (group E);hydrogen-rich culture medium + endotoxin group (group HE).The cells were cultured in high-glucose DMEM culture medium in group C.The cells were incubated in hydrogen-rich culture medium containing hydrogen 0.6 mmol/L in group H.The cells were incubated in highglucose DMEM culture medium containing 50 μg/ml lipopolysaccharide in group E.The cells were incubated in hydrogen-rich culture medium containing 50 μg/ml lipopolysaccharide and 0.6 mmol/L hydrogen in group HE.Transepithelial electrical resistance (TEER) was measured before incubation or culture,and at 6,12 and 24 h of incubation or culture.The viability of Caco-2 cells was measured by methyl thiazolyl tetrazolium assay at 24 h of incubation or culture.The expression of ZO-1 mRNA in Caco-2 cells was determined using real-time reverse transcriptase polymerase chain reaction at 6,12 and 24 h of incubation or culture.The distribution of ZO-1 in Caco-2 cells was observed by immunofluorescence at 24 of incubation or culture.Results Compared with group C,TEER was significantly decreased at 6,12 and 24 h of incubation or culture,and the expression of ZO-1 mRNA was significantly down-regulated in E and HE groups (P＜0.05),and no significant change was found in the parameters mentioned above in group H (P＞0.05).Compared with group E,TEER was significantly increased at 6,12 and 24 h of incubation or culture,and the expression of ZO-1 mRNA was significantly up-regulated in group HE (P＜0.05).The distribution of ZO-1 protein in cell membrane became discontinuous,and the distribution of ZO-1 protein in cytoplasm was significantly increased in group E.Compared with group E,the distribution of ZO-1 protein in cell membrane was significantly increased and gradually became continuous,and the distribution of ZO-1 protein in cytoplasm was significantly decreased in group HE.Conclusion The mechanism by which hydrogen reduces the damage to human colon epithelial cell barrier is related to up-regulation of ZO-1 expression and improvement in the redistribution of ZO-1 protein.