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.

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 investigate the role of Rho kinase (ROCK) in the protective effects of hydrogen on intestinal epithelial barrier function in sepsis. Methods Caco-2 cells were cultured routinely, and divided into 6 groups randomly (n=3):control group (C group), hydrogen-rich medium group (H group), lipopolysaccharide (LPS)-treatment group (L group), hy?drogen+LPS-treatment group (HL group), Rho kinase inhibitor (Y-37632) treatment group (Y group) and Rho kinase inhibi?tor Y-27632+LPS-treatment group (YL group). H group was treated with 0.6 mmol/L hydrogen-rich media. The concentra?tion of LPS and Y-27632 were 50 mg/L and 25μmol/L separately. After the Caco-2 monolayer model was established, the transepithelial electrical resistance (TEER) values were measured regularly. When the TEER value reached 800Ω·cm2, the treatment was administered. Then TEER values were measured at 6 h, 12 h and 24 h, and FITC-dextran permeability was de?tected at 24 h. Cells were seeded on 6-well plates. After cell density reached 80%-90%, treatments were given randomly. The real time-polymerase chain reaction (RT-PCR) was conducted to assess mRNA levels of ZO-1 and ROCK mRNA. ZO-1 and ROCK protein expression levels were detected by Western blot assay. Results Compared with C group, TEER values were elevated in 12 h and 24 h in H group (P<0.05). There were no statistical significances in FITC-dextran permeability,
protein expression levels of ZO-1 and ROCK between C group and H group (P>0.05). TEER values were elevated at 6 h, 12 h and 24 h in Y group (P<0.05). There was no significant difference in FITC-dextran permeability between C group and Y group (P > 0.05). The mRNA expression of ZO-1 increased and mRNA expression of ROCK decreased in Y group (P <0.05). The TEER values reduced at 6 h, 12 h and 24 h in L group. The FITC-dextran permeability increased significantly, mRNA and protein expressions of ZO-1 significantly decreased, mRNA and protein expressions of ROCK significantly in?creased in L group (all P<0.05). Compared with L group, TEER values increased significantly at 6 h, 12 h and 24 h in YL group, FITC-dextran permeability decreased, mRNA expressions of ZO-1 increased, mRNA expressions of ROCK de?creased in YL group (P<0.05). Compared with L group, TEER values increased at 6 h, 12 h and 24 h in HL group, FITC-dextran permeability reduced markedly, protein expressions of ZO-1 increased at each time point, protein expressions of ROCK decreased at each time point in HL group (P<0.05). Conclusion Hydrogen can protect intestinal barrier function against sepsis, ameliorate the integrity and permeability of intestinal epithelium and increase the expressions of intercellular tight junction proteins. The suppression of Rho kinase over-expression induced by LPS may be involved in these protective effects of hydrogen.

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 establish a mouse model of orthotopic Lewis lung carcinoma using Matrigel, to evaluate the tumor growth and metastasis, and to provide a more stable mouse model of orthotopic lung cancer, which is more similar to human lung cancer.Methods Logarithmic phase of cultured Lewis lung cancer cells were suspended in Matrigel, vac-cinated into the left lung of inbred C57BL/6 mice.Five mice were killed on the 4th, 7th, 10th, 13th, and 16th days, re-spectively, and to observe the median survival, tumor formation rate, tumor growth, and metastasis.Pathological changes of the mouse lung, liver, kidney and spleen were examined.Results In 5 mice killed on the 7th postoperative day, small tumor nodules were observed on the lung in three mice and no tumor was visible by gross inspection in the other two mice, but small tumor nodules were observed under the microscope.For all the mice killed on the 10th postoperative day, tumors were visible to the naked eye on the lung of all the five mice.On the 13th day, orthotopic tumor was observed on the lung with bloody pleural effusion and pleural metastasis in all the five mice.On the 25th day, in addition to the pleural metasta-sis, one mouse had pericardial metastasis and renal metastasis.The survival periods of the 5 mice were 17 d, 20 d, 22 d, 22 d, and 25 d, respectively, with a median survival period of 21.2 d (17-25 d), and the tumor formation rate was 100%.Conclusions Mouse models of orthotopic Lewis lung carcinoma is successfully established using injection of tumor cells suspended in Matrigel.This model is more similar to the growth of human lung cancer, with good stability, high tumor formation rate and characteristics of distant metastasis, therefore, is worthy of further 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.

Objective To investigate the expression and clinical significance of silent mating type information regulation 2 homolog-1(SIRT-1)and nuclear factor-κB(NF-κB)in non-small cell lung cancer (NSCLC). Methods The expressions of SIRT-1 and NF-κB were evaluated by immunohistochemistry in 108 selected cases of primary NSCLC and 48 samples of para-carcinoma normal tissue. The relationships of the expressions of SIRT-1 and NF-κB and clinical pathological features were analyzed,respectively. Results Immunohistochemical results showed that the positive expression rates of SIRT-1 and NF-κB in NSCLC tissue were 90. 7%(98 / 108)and 94. 4%(102 / 108),respectively,significantly higher than those in normal lung tissue 4. 2%(2 / 48),16. 7%(8 / 48),χ2 = 108. 237,P = 0. 000;χ2 = 96. 683,P = 0. 000,and the expre-ssion of SIRT-1 and NF-κB showed a positive correlation(r = 0. 480,P = 0. 001). There were significant posi-tive correlations between the expression of SIRT-1 and tumor size(r = 0. 227,P = 0. 018),TNM stage(r =0. 298,P = 0. 002)and lymph node metastasis(r = 0. 280,P = 0. 003),and there was negative correlation between the expression of SIRT-1 and differentiation(r = - 0. 300,P = 0. 002),and there were no correlation between the expression of SIRT-1 and sex,age and histological type in NSCLC tissues. There were significant positive correlation between the expression of NF-κB and TNM stage(r = 0. 256,P = 0. 009)and lymph node metastasis(r = 0. 261,P = 0. 006),and there was negative correlation between the expression of NF-κB and differentiation(r = - 0. 235,P = 0. 013),and there were no correlation between the expression of NF-κB and sex,age,histological type and tumor size in NSCLC tissues. Conclusion The positive expression rates of SIRT-1 and NF-κB in NSCLC tissue are significantly higher than those in normal lung tissue,and they are rela-ted to TNM stage,lymph node metastasis and differentiation,and the former is also related to tumor size. High expression of SIRT-1 and NF-κB may play important roles in the occurrence and development of NSCLC.