BACKGROUND:Brahma-related gene 1 (Brg1), a catalytic subunit of an important chromatin remodeling complex, has been considered as a key nuclear transcriptional factor, and tends to be decreased in diabetic cardiomyopathy.
OBJECTIVE:To construct an adenovirus vector carrying Brg1, and observe its protective role in oxidative stress induced-cardiomyocyte apoptosis.
METHODS:The recombinant adenovirus plasmid was linearized and transfected into HEK293 cel s using Fugene HD for packaging and amplification. The adenovirus particles were further purified, quantified, and sequential y transfected to cardiomyocytes of neonatal Sprague-Dawley rats. The Adeno-EGFP transfected and non-transfected cardiomyocytes were used as control group. 24 hours later, the transfection efficiency was observed by fluorescent microscope, and expressions of Brg1 mRNA and protein were detected by quantified PCR and western blotting. After treatment with 100 μmol/L H2O2 for 12 hours, the expressions of Brg1 protein and cleaved-Caspase 3 were measured by western blotting, and cel apoptosis was analyzed by flow cytometry.
RESULTS AND CONCLUSION:(1) The recombinant adenovirus vector of Brg1 had been successful y transfected into cardiomyocytes with higher expressions of Brg1 mRNA and protein, and the transfection efficiency reached more than 90%. (2) After H2O2 treatment, the Brg1 was significantly down-regulated in contrast to the up-regulation of cleaved-Caspase 3;the flow cytometry data showed that the apoptotic cel s were increased. But in Adeno-Brg1 transfected cardiomyocytes, the H2O2 induced cel apoptosis was significantly decreased compared with non-transfected cel s and empty vector transfected cel s. (3) These results suggest that oxidative stress can directly inhibit the Brg1 expression, and overexpression of Brg1 can protect the cardiomyocytes from cel apoptosis induced by oxidative stress.

Objective To observe the effect of Oxycontin combined with Gabapentin for treatment of malignant neuropathic pain.Methods Sixty-three cases of malignant neuropathic pain in Jinshan Hospital Affiliated to Fudan University were randomly divided into group A, B and C.Patients of which were given Oxycontin, Gabapentin, Oxycontin combined with Gabapentin respectively for pain treatment.The analgesic effects, toxic reaction side effects, quality of life, and immune function were all compared in three groups.Results Compared with pretherapy, the cancer pain score (NRS), quality of life (QOL) and karnofsky performance status(KPS) scores in all groups were changed significantly after drugs therapy(F=375.852,154.612, 151.838,P＜0.05).The levels of CD3,CD4, CD4/CD8 and NK cells in all groups were higher than before therapy(F=158.935,108.145,366.973,92.090,P＜0.05).After treatment,the NRS, QOL and KPS scores in group C were 2.00± 0.86,44.80± 6.07, 84.50± 6.05, in group A were 3.35 ± 0.67,37.35 ± 5.71,74.50 ±10.99,and in group B were 4.05±0.94,35.85±5.90,72.00±8.34, and the different were significant (F =3.250,10.499,3.465,P＜0.05).The levels of CD3, CD4, CD4/CD8and NK cells in group C were (72.94 ±5.63)％,(41.52±4.19)％, 1.86±0.30, (27.57±6.86)％,in group A were (62.84±5.27)％, (33.84 ±5.40)％,1.35±0.37, (20.49±6.67) ％,and in group B were (62.22±8.10)％, (33.19±6.90)％, 1.32 ± ±0.41, (20.32±5.63) ％, and the different were significant (F =3.377,3.344,3.352,3.386, P＜ 0.05).The patient in group C had less adverse effects than those in group A and B.Conclusion Oxycontin and Gabapentin in treatment of malignant neuropathic pain is effective.

Background and purpose:As the second general platinum antineoplastic agent, carboplatin has been applied in treating more and more malignant tumors clinically. But it has severe bone marrow suppression when applied in large doses. The purpose of this research was to explore the influence of carboplatin of fractional dose, combined with X ray,on the immunity of Lewis lung cancer in mice. Methods:The model of tumor-bearing mice was induced by injecting Lewis lung cancer cells into the right infra-axillary dermis. In cases given the same total dose, the tumor growth and the immune suppression effect of carboplatin of single dose or of fractional dose , combined with X ray radiation, on the mouse with Lewis lung cancer were observed. Results:The concentration of IL-10 and TNF-? in serum,the spleen exponent were respectively(144.9?48.4)ng/ml,(194.63?64.4)ng/ml,(12.07?2.2)mg/ g in the control group;(279.0?46.9)ng/ml,(71.5?8.4)ng/ml,(5.52?1.31)mg/g in carboplatin of single dose with X ray radiation group, which ,compared with the control group, can dramatically increase the concentration of IL-10,decrease the concentration of TNF-? in the serum ,and dramatically decrease the spleen exponent(P

Objective To investigate the relationship between the effect of unmethylated cytosine phosphate-guanine oligodeoxynucleotide ( CpG ODN ) 7909 on X-ray-induced G2/M phase arrest and apoptosis and the phosphorylation of ATM kinase.Methods Human lung adenocarcinoma A549 cells were randomly classified into five groups,control,CpG group,irradiation group,CpG + irradiation group and ATM siRNA + CpG + irradiation group.Cell survival fraction was evaluated by clonogenic assay.Cell cycle and apoptosis were analyzed by flow cytometry.The expressions of ATM,checkpoint kinase-2(Chk2) and p53 were detected with Western blot.Results Compared to the situation under X-ray irradiation alone,decreased cell clonogenic survival,prolonged G2/M arrest,and increased cell apoptosis were observed after the combination treatment with CpG ODN7909 and X-rays ( t =13.41,17.32 and 7.71,P ＜ 0.05).Moreover,the phosphorylations on ATM,Chk2,and p53 were increased in the irradiated A549 cells that had been pre-treated with CpG ODN7909.After ATM siRNA interfering,abrogation of G2/M arrest,reduction of apoptosis,and decrease of Chk2 and p53 phosphorylation were found in A549 cells treated with CpG ODN7909 and X-rays ( t =26.84,2.98,47.24 and 67.47,P ＜0.05).Conclusions CpG ODN7909 can enhance the X-ray-induced phosphorylation of ATM kinase in human lung adenocarcinoma cells in vitro,which might be involved in regulating G2/M phase arrest and apoptosis.

Objective To explore the combination effect of unmethylated cytosine-phosphate-guanine oligodeoxynucleotide (CpG ODN) 1826 and X-rays on Lewis lung cancer in mouse and the dose response of CpG ODN.Methods The tumor-bearing mouse model was established by injecting Lewis lung cancer cells into the right infra-axillary dermis of mouse.Sixty-four C57BL /6 J mice were evenly randomized into eight groups with 8 mice each:control group,IR group,CpG OND1826 0.15 mg group,CpG OND1826 0.3 mg group,CpG OND1826 0.45 mg group,CpG OND1826 0.15 mg + IR group,CpG OND1826 0.30 mg+ IR group,and CpG OND1826 0.45 mg + IR group.On the 1st,2nd,and 9th days,CpG ODN was injected into mouse.After 3 hours of injection,the mice were start to irradiate with X-rays once a day on the 2nd-6th days,and the total dose was 12.50 Gy.Tumor growth and TGD were measured,and the apoptosis of tumor cells were examined with TUNEL.Results The Lewis lung cancer-bearing model was successfully established in all mice.Under the treatments of CpG OND1826 and irradiation,the tumor volumes were smaller than that of control group,and the tumor volumes of CpG OND1826 0.45 mg+IR group was the smallest.TUNEL results revealed that the apoptosis rate were (2.40 ± 0.51 )％ in control group,(5.62 ±0.50)％ in IR,(7.13±0.83)％ in CpG OND1826 0.15 mg,(11.63±1.06)％ in CpG OND1826 0.3 mg,(19.13 ±0.83)％ in CpG OND1826 0.45 rag,( 12.88±0.83)％ in CpG OND1826 0.15 mg+ IR,(20.57±2.37)％ in CpG OND1826 0.3 mg+ IR,and (28.17 ±3.31)％ in CpG OND1826 0.45 mg + IR group,and thus the apoptosis rate of every therapy group was higher than that in control ( t=11.15,7.91,17.82,39.48,24.73,16.61 and 17.05,P＜0.05).The apoptosis rates of CpG ODN1826 plus X-ray irradiation group were significantly higher than those in IR alone ( t =13.78,15.08 and 17.47,P＜0.05 ) or CpG ODN group (t=18.53,9.66and7.51,P＜0.05).Conclusions CpG ODN1826 can dramatically increase the efficiency of radiotherapy by inhibiting tumor growth and promoting lumor apoptosis.

Purpose To investigate the relationship between the expression of Runx3 protein and pathogenesis, development,clinicopathological features and its prognostic significance in gastric adenocarcinoma.Methods Immunohistochemical SP method was used to detect the expression of Runx3 protein in 63 cases of gastric cancer,19 cases of atypical hyperplasia and 10 cases of normal gastric mucosa obtained from patients whose partial gastrotomy was performed for benign diseases.Results Compared with gastric atypical hyperplasia tissue (68.4%) and normal gastric mucosa (90%), the positive rate of Runx3 protein in gastric cancer (39.7%) was significantly lower (P＜0.05); the expression of Runx3 protein was related to tumor differentiation, invasive depth and lymphnode metastasis, but not to age, sex, tumor size and TNM stages; the survival rate of the patients with Runx3 protein expression was higher than that without Runx3 expression (P＜0.05).Conclusion Runx3 protein may play an important role in the occurrence and development of gastric cancer, and it is an important marker in evaluating the prognosis of the patients.

Objective To study the effects of 6-week aerobic treadmill exercise on the genes expression in skeletal muscle of C57BL/6 mice.Gene array was performed and the data was analyzed to evaluate the adaptive changes at gene level caused by exercise.Methods Twenty male C57BL/6 mice were randomly divided into two groups:control group(C) and aerobic exercise group (E).The mice from group E were forced to run on treadmill for 6 weeks.After 6 weeks,all mice were fasted for 16 hours,and then sacrificed.Their flexor muscle was removed.4 samples from each group were analyzed with gene chips to dissect the differentially expressed genes between the two groups.Results Our results showed that 723 genes were differentially expressed in E group as compared with C group after the 6 weeks aerobic exercise,among which 510 genes were up-and 213 down-regulated.There were 6 differentially expressed calcium signaling genes and 2 ofwhich(Gnall and Pdgfra) were up-regulated,while the others(Phka1,Picd4,RYR1 and Ppid)were down regulated.

Objective To evaluate the therapeutic effect and side effects of three-dimensional conformal re-irradiation combined with nedaplation plus 5-fluorouracil synchronously in treatment of recurrent esophageal carcinoma.Methods Fifty-four patients with recurrentesophageal carcinoma were randomly divided into two groups:study group with 27 patients who were treated with three-dimensional conformal re-irradiation (50 ～ 56Gy/25 ～ 28 F) combined with chemotherapy synchronously and control group with 27 patients who were treated with re-irradiation (50 ～56 Gy/25 ～ 28 F)combined with chemotherapy orderly.The patients of study group were treated with nedaplation80 mg/(m2 · d)plus5-FU 350 mg/( m2 · d),from 1 to 5 day synchronously in radiotherapy and after radiotherapy.The patients of control group were treated with nedaplation80 mg/( m2 · d)plus 5-FU 350 mg/(m2 · d)d from 1 to 5 day orderly after radiotherapy.The patients were treated every 3 -4weeks as a cycle with total 2 - 4 cycles in these two groups.Results ( 1 ) The short-term effect:the short-term effect(CR + PR)of the two groups were 59.2％ (16/27) and 55.5％ (15/27) respectively,there was no statistical difference between the two groups( x2 =0.076,P =0.5 ).( 2 ) The survival rate:the survival rate of one,two and three years were 54.2％,26.4％,15.7％ in study group and 40.3％,19.7％,10.8％ in control group respectively The survival rate in study group was significantly higher than that in control group ( x2 =6.87,P =0.032 ).( 3 ) The study of side effects:there was significant difference on the hematologic toxicity between the study group and the control group ( x2 =5.882,P =0.043 ).And there was no significant difference on the non-hematologic toxicity between the two groups ( x2 =4.25,P =0.75 ).(4) The acute radiationesophagitis:There was no statistical difference on radiation-esophagitis between the two groups ( x2 =0.869,P =0.661 ).Conclusion Three-dimensional conformal re-irradiation combined with nedaplation plus 5-fluorouracil synchronously for recurrent esophageal carcinoma has a good efficacy and can also improve the survival rate.

AIM:To probe whether CpG oligodeoxyribonucleotides 7909 (CpG ODN7909) combined with Toll like receptor (TLR)9 affected the chemotherapeutic sensitivity of doctaxel (DOC) in human lung cancer A549 and H520 cell lines.METHODS:Sequences of TLR9 siRNAs were designed.A549 and H520 cells were transfected with TLR9 siR-NA by lipofectamine.The expression of TLR9 was detected by Western blot .The cell activity was measured by CCK-8 as-say.The experiments were divided into blank control group , control siRNA group and TLR9 siRNA interference group.The cell cycle distribution and cell apoptosis were analyzed by flow cytometry .The expression of P38 and Bax was determined by Western blot .The cells in each group were exposed to CpG ODN 7909 and/or DOC.RESULTS: In A549 cells and H520 cells, CpG ODN7909 alone had no obvious effect on the cell activity , G2/M phase arrest and apoptosis , but in-creased the protein expression of P 38 and Bax ( P<0.01) .In addition, there was no significant changes of the above inde-xes in CpG ODN7909 treated-TLR9 siRNA group was observed .DOC alone significantly inhibited the cell activity , higher the G2/M phase fractions, apoptotic rates and Bax expression (P<0.01), but didn’t affect the expression of P38 in all 3 groups.Compared with the cells treated with DOC alone , the cells treated with CpG ODN7909 combined with DOC exhibi-ted lower cell activity, higher G2/M phase fractions, apoptosis rates and more Bax expression (P<0.01), showed no sig-nificant change of P38 expression.In addition, there was no significant change of the above indexes in CpG ODN 7909 com-bined with DOC treated-TLR9 siRNA group was observed .CONCLUSION:CpG ODN7909 may enhance the chemothera-peutic sensitivity of DOC in human lung cancer cells by combining with TLR 9.The mechanism might be related to enhan-cing the inhibitory effect and apoptosis of DOC on the cell activity in vitro, arresting the cells at G 2/M phase of the cells .

Objective Identify novel protein kinase Cε(nPKCε)-interacted proteins in the cortex of hypoxic preconditioned mice.Methods Immunoprecipitation (IP) and two-dimensional electrophoresis (2-DE) combining with ImageMaster 2D Platinum software were applied to analyze the differential expressions of nPKCe-interacted proteins;the target protein spots were identified by matrix-associated laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and Western blot.Results Compared with control group,there were 34 upregulated protein spots and 20 downregulated protein spots in cytosolic fraction,while 27 upregulated prtein spots and 28 downregulated protein spots were determined in particulate fraction of cerebral cortex of HPC mice.The levels of nPKCε-interacted HSP 70 and 14-3-3γ/protein expressions increased significantly in both cytosolic and particulate fractions;but the protein level of nPKCε-interacted HSP60 increased only in particulate fraction of cerebral cortex of HPC mice.Conclusion nPKCε might be involved in the development of cerebral HPC via the regulation of its interacted proteins such as HSP60,HSP70 and 14-3-3γ.