Objective To investigate the effect of kaempferol(KAE)on the function of drug-resistant Bel-7402/5-Fu cells.Methods Bel-7402/5-Fu cells were treated with KAE,and cells were divided into the control group and the drug group(0.064,0.320,1.600,8,40,200 μmol/L KAE).Cells were divided into the si-NC group and the DNA-PKcs interference group,or the control group,the KAE group,the KAE+si-DNA-PKcs group or the KAE+DMSO group,the KAE+MG132 group and the KAE+CQ group based on interfering DNA dependent kinase catalytic subunits(DNA-PKcs)or addition of proteasome inhibitor MG132 or autophagy inhibitor CQ.Cell proliferation was detected using CCK-8.The expression level of histone H2AX phosphorylation(γ-H2AX),DNA-PKcs,DNA double strand break repair/V(D)J recombinant protein(Artemis)and drug pump gene(P-gp)were analyzed using real-time fluorescence quantitative PCR(RT-qPCR)and Western blot assay.Cell cycle and apoptosis were detected by flow cytometry.The stability of DNA-PKcs proteins was analyzed by protein stability experiments.Ubiquitination of DNA-PKcs protein was evaluated by immunoprecipitation assay.Results Compared to the control group,treating cells with 8 μmol/L KAE for 24 h inhibited about 50%of cell proliferation ability.Therefore,this time and concentration were chosen for subsequent research.Compared to the control group,the expression level of γ-H2AX mRNA and protein significantly increased,while expression levels of DNA-PKcs,Artemis and P-gp mRNA and proteins significantly decreased in the KAE group(P＜0.05).Compared to the control group,KAE promoted cell cycle arrest in the G2/M phase of Bel-7402/5-Fu cells and increased cell apoptosis.Compared to the si-NC group,siRNA-1664 significantly downregulated the mRNA and protein expression levels of DNA-PKcs(P＜0.05).Compared with the KAE group,the effect of KAE was further promoted in the KAE+si-DNA-PKcs group of Bel-7402/5-Fu cells.Compared with the control group,the protein expression level of DNA-PKcs decreased in the KAE+DMSO group(P＜0.05).Compared with the KAE+DMSO group,the protein expression level of DNA-PKcs increased in the KAE+MG132 group(P＜0.05),while there was no significant change in the protein expression level of DNA-PKcs in the KAE+CQ group(P＞0.05).Compared to the control group,there was promoted ubiquitination of DNA-PKcs in the KAE+DMSO group,and the inhibited ubiquitination in the KAE+MG132 group(P＜0.05).Conclusion KAE may induce cell apoptosis and cell cycle arrest in drug-resistant Bel-7402/5-Fu cells.

Objective To explore the effect of non-invasive positive pressure ventilation (CPAP) on airway inflammation factors of patients with chronic obstructive pulmonary disease (COPD).Methods A total of 60 patients with COPD were randomly divided into control group (n =30) and observation group (n =30).The patients in the control group were treated with conven-tional symptomatic support.In the observation group,non-invasive positive pressure ventilation was applied on the basis of the control group.IL-8 and TNF-α were measured by enzyme-linked immunosorbent assay.The cell suspension,neutrophil percentage of white blood cells,clinical efficacy and the airway inflammatory factors were compared between two groups.Results After treatment,the levels of pH and p (O2) in the observation group were significantly higher than those in the control group (P ＜ 0.05),and the level of p (CO2) were significantly lower than the control group (P ＜ 0.05).There was no significant difference in neutrophil count before and after treatment between the two groups (P ＞ 0.05).The IL-8 and TNF-α in the observation group were significantly lower than those in the control group (P ＜ 0.05).The incidence rate of complications in the observation group was 3.33％,which was significantly lower than 10.00％ in the control group (P ＜ 0.05).Conclusion Combination of non-invasive positive pressure ventilation and conventional therapy is effective in treatment of patients with COPD.

Objective To explore the effect of non-invasive positive pressure ventilation (CPAP) on airway inflammation factors of patients with chronic obstructive pulmonary disease (COPD).Methods A total of 60 patients with COPD were randomly divided into control group (n =30) and observation group (n =30).The patients in the control group were treated with conven-tional symptomatic support.In the observation group,non-invasive positive pressure ventilation was applied on the basis of the control group.IL-8 and TNF-α were measured by enzyme-linked immunosorbent assay.The cell suspension,neutrophil percentage of white blood cells,clinical efficacy and the airway inflammatory factors were compared between two groups.Results After treatment,the levels of pH and p (O2) in the observation group were significantly higher than those in the control group (P ＜ 0.05),and the level of p (CO2) were significantly lower than the control group (P ＜ 0.05).There was no significant difference in neutrophil count before and after treatment between the two groups (P ＞ 0.05).The IL-8 and TNF-α in the observation group were significantly lower than those in the control group (P ＜ 0.05).The incidence rate of complications in the observation group was 3.33％,which was significantly lower than 10.00％ in the control group (P ＜ 0.05).Conclusion Combination of non-invasive positive pressure ventilation and conventional therapy is effective in treatment of patients with COPD.

Objective:To study influence of renal sympathetic denervation (RDN)on cardiac function of dogs with heart failure (HF).Methods:A total of 40 dogs were randomly and equally divided into RDN group [received bilat- eral renal artery radiofrequency ablation (RFA)]and model group (only received femoral puncture).Pacemaker was implanted in every dog,and dog HF model was established using rapid right ventricular pacing.Cardiac and re-nal function indexes,BNP and sympathetic activity index levels were observed and compared between two groups be- fore RFA/sham operation,instant and four weeks after model establishment.Results:After operation four weeks, compared with model group,there were significant reductions in levels of epinephrine (E)[(362.69±42.54)ng/ml vs.(290.36±42.32)ng/ml],renin (R)[(305.46± 39.68)ng/ml vs.(230.04±32.80)ng/ml],aldosterone (AD)[(408.00±38.56)ng/ml vs.(246.00± 48.37)ng/ml],angiotensin Ⅱ (ATⅡ)[(280.00±48.08)pg/ml vs.(172.00±25.04)pg/ml]and norepinephrine (NE)[(425.65±50.54)ng/ml vs.(316.76±46.29)ng/ml]in RDN group (P<0.05 all);there were significant reductions in HR,respiratory rate (RR)and BNP level in RDN group,P<0.05 all;there were significant rise in SBP,LVEF,CO,CI,left ventricular pressure maximal rising rate (+dp/dtmax),left ventricular pressure maximal dropping rate (-dp/dtmax)and left ventricular end-systolic pressure (LVESP),and significant reductions in left ventricular end-systolic dimension (LVESd),left ventricular end-diastolic dimension (LVEDd)and left ventricular end-diastolic pressure (LVEDP)in RDN group,P<0.05 all.Conclusion:RDN can decrease renal sympathetic activity,improve heart function,inhibit myocardial remode- ling,its therapeutic effect is significant

Objective To evaluate the effects of Ro20?1724 on cognitive dysfunction induced by repetitive ketamine anesthesia in immature rats. Methods Thirty?two Sprague?Dawley rats of both sex, aged 21 days, weighing 45-55 g, were randomly divided into 4 groups ( n=8 each ) using a random number table: control group ( group C ) , ketamine group ( group K ) , ketamine+Ro20?1724 group ( group K+R) , and ketamine+anhydrous alcohol group ( group K+A) . In K, K+R and K+E groups, 70 mg∕kg ketamine was intraperitoneally injected once a day for 7 consecutive days. The equal volume of normal saline was given instead in group C. Two days after the rats were fed a common diet, 0.5 mg∕kg Ro20?1724 and the equal volume of anhydrous alcohol were injected in K+R and K+E groups, respectively, and the equal volume of normal saline was given instead in K and C groups, once a day for 7 consecutive days. Morris water maze test was used to test cognitive function, and the escape latency and frequency of crossing the original platform were recorded. The rats were sacrificed after the end of behavior tests, and hippocampi were removed to detect the content of brain?derived neurotrophic factor ( BDNF) in CA1 region using enzyme?linked immunosorbent assay. Results Compared with group C, the escape latency was significantly prolonged on 1st-4th days in K and K+E groups, the escape latency was prolonged on 3rd-4th days in K+R group, and the frequency of crossing the original platform and content of BDNF in CA1 region were decreased in K, K+R and K+E groups. Compared with K group, the escape latency was significantly shortened, and the frequency of crossing the original platform was increased on 3rd-4th days, and the content of BDNF in CA1 region was increased in K+R group, and no significant changes were found in the parameters mentioned above in K+E group. Conclusion Ro20?1724 can improve cognitive dysfunction induced by repetitive ketamine anesthesia in immature rats, and enhanced production of BDNF is involved in the mechanism.

Objective To investigate the influence of the serum vascular endothelial growth factor (VEGF) in the glioma patients by the malignant degress.Methods Quantitative analysis the serum VEGF levels with the double antibody sandwich assay (ELISA) for 35 cases of glioma patients and 10 cases of healthy human and analyze the correlation of them.Results Mean serum VEGF expression in glioma patients was 120.71 ±45.99 pg/ml,normal value of the healthy people was 63.70 ± 6.50,the difference was statistically significant (P ＜0.05); serum VEGF expression was 156.43 ± 14.90 pg/ml in high-grade glioma patients (Ⅲ-Ⅳ grade) and 67.14 ± 6.12 pg/ml in low-grade glioma patients (Ⅰ-Ⅱ grade),the difference was statistically significant (P ＜ 0.05) ; there' s no different from low-grade glioma patients (Ⅰ-Ⅱ grade) and nomal(P ＞ 0.05).Conclusion The serum VEGF levels in the glioma patients was influence by the malignant degress.

Objective: To observe influence of long-term exercise training on carotid intima-media thickness (IMT) in patients with mild-moderate hypertension. Methods: A total of 92 patients with essential mild-moderate hypertension were randomly divided into routine treatment group (n=46) and exercise group (n=46, received exercise training based on routine treatment) according to number table. All patients were followed up for one year, and the blood pressure, carotid artery diameter and carotid IMT were measured, compared and analyzed in two groups before and after treatment. Results: There was no significant difference in systolic and diastolic blood pressure before treatment between two groups (P＞0.05). Compared with baseline value, there were significant decrease in systolic blood pressure [(176.66±l1.78)mmHg vs. (130.89±13.01)mmHg], diastolic blood pressure [(101.43±6.41)mmHg vs. (81.71±8.45)mmHg], carotid artery diameter [(6.62±0.97)mm vs. (6.22±1.01)mm] and carotid IMT [(0.98±0.12)mm vs. (0.84±0.11)mm] in exercise group after one-year training (P<0.05 all), and they were all significantly lower than those of routine treatment group (P<0.05 all ). Conclusion: Long-term exercise training can effectively control blood pressure, decrease carotid artery diameter and carotid intima-media thickness.

ObjectiveTo investigate the expression and biological significance of PIM1 in the breast cancer,atypical hyperplasia breast,and normal breast tissues.MethodsThe protein expression levels of PIM1 in the breast cancer,atypical hyperplasia breast,and normal breast tissues were assayed by immunohistochemistry and Western blot.ResultsThe expression rates of PIM1 protein were 78.75％ (63/80)in breast cancers,42.00％ (21/50) in atypical hyperplasia tissues,and 23.33 ％ (7/30) in normal breast tissues,respectively.A significant correlation was found between PIM1 expression and the clinical stage and lymphonodus metastasis.However,no significant correlation was found between PIM1 expression and patients'age and tumor sizes.The level of PIM1 protein in breast cancers was obviously higher than that in non - cancerous tissues.ConclusionsThe positive rate of PIM1 in breast cancers is significantly higher than that in atypical hyperplasia breast tissues and in normal breast tissues.PIM1 may be an early molecularevent in mammary tumorigenesis,and its overexpression may significantly relate to the malignant progression.It would be a new parameter for the early diagnosis and a biomarker for breast cancers.It is feasible to utilize paraffin specimens for index test with advantages of convenience,easy availability,and low expense.

Objective To investigate the blood pressure circadian rhythm in patients with IgA nephropathy by ambulatory blood pressure monitoring and explore its role in the disease progression. Methods A cross sectional study was carried out.Blood pressure rhythm was studied by ambulatory 24-hour monitoring with a portable oscillometric recorder in selected patients with primary IgA nephropathy.The term dipper was described as blood pressure during night dropped at least 10％ below daytime blood pressure.The term non-dipper referred to those in whom the nocturnal decline in blood pressure was less than 10％.Clinicopathological indices between dipper and non-dipper groups were compared. Results Ninety-three patients completed ambulatory blood pressure monitoring among whom 68 (73％) patients were non-dipper.The frequency of non-dipper was 70％,70％ and 81％ in the patients at chronic kidney disease stage 1,2 and 3 or more.The frequency did not differ among these three group patients (P=-0.587).77％ of patients with hypertension and 69％ of patients with normotension were non-dipper (P=0.373).The disappearance of blood pressure circadian rhythm in IgA nephropathy was not influenced by age,gender,blood pressure,proteinuria,renal function and renal pathology lesions.Among the patients who were followed up regularly for more than 12 months (n=54),patients in the dipper group had a trend of slower eGFR decline rate than those in non-dipper group albeit the difference was not significant (P=0.329).Subgroup analysis revealed that in patients with hypertension and non-dipper (n=29),the eGFR decline rate was much faster than that in dipper group[(-6.79±11.58 )vs (-0.34±1.74) ml ·min-1 ·(1.73 m2)-1·year-1,P=0.019]. Conclusions Most patients with IgA nephropathy present disappearance of blood pressure circadian rhythm,even among those at an early stage or without hypertension.The loss of blood pressure rhythm may be associated with a rapid renal function decline rate in those with hypertension.

Objective To discuss the diagnosis and surgical management of multilocular cystic renal cell carcinoma (MCRCC) and to evaluate the gene function of the mutation of von Hippel-Lindau (VHL) gene in MCRCC. Methods Seventeen MCRCC cases (11 men and 6 women) out of 512 cases of renal cell carcinoma from 2000 to 2010 were retrospectively analyzed. The mean age of the 17 patients was 46 years (37-61 years). Ultrasonography and CT were available in all 17 cases, and 1 case was misdiagnosed as parapelvic renal cyst. The mutation of VHL gene was detected by PCR in the specimens of can-cerous tissue and adjacent normal tissue from 11 cases of MCRCC. Results Three of 17 cases underwent nephron sparing surgery, the others underwent radical nephrectomy. One case underwent unroofing of parapelvic renal cyst, but the rapid frozen pathology of the cyst wall showed renal cell carcinoma of clear type. As a result, radical nephrectomy was eventually performed. All 17 cases were confirmed as MCRCC by eva-luating pathological characteristics, such as the cyst wall lined by single or several layers of clear tumor cells and the nuclei which were small and anachromasis. Clinical stages of all cases were T1N0M0, in which there were 14 cases with pathological T1G1 and 3 cases with pathological T1G2. All patients underwent a follow-up of 9 to 36 months (mean, 12 months) without recurrence or metastasis. Mutation of VHL gene was detected in 7 of 11 cases (64%), but all adjacent normal tissues were negative. Conclusions As a rare subtype of renal cell carcinoma, MCRCC is difficult to diagnose. CT is an essential measure in diagnosis of MCRCC preoperatively. Because of the good prognosis of reported cases, nephron sparing surgery for the treatment of MCRCC is recommended. VHL gene mutations may play an important role in the carcinogenesis of MCRCC.