Objective To investigate the therapeutic effect of ipratropium bromide on bronchial spasm of patients with asthma and chronic obstructive pulmonary disease (COPD).Methods 174 patients with asthma and COPD were selected and divided into two groups, 87 cases in the control group treated with budesonide and formoterol fumarate powder for inhalation , 87 cases in the experimental group received ipratropium bromide on the basis of the control group, pulmonary ventilation function, airway resistance and cytokine levels in induced sputum, the clinical effect and incidence of adverse reactions were compared after the treatment.Results The effective rate in the control group(81.61%)was lower than the experimental group (93.11%), with significant difference (P<0.05); compared with the control group, levels of peak expiratory flow (PEF), maximum midexpiratory flow (MMEF), forced expiratory volume in one second to forced vital capacity ratio (FEV1%), forced vital capacity (FVC) were higher in the experimental group after treatment, levels of airway resistance (Raw), Raw% were lower, level of specific airway conductance (Gsp) was higher after treatment, induced sputum levels of vascular endothelial growth factor (VEGF), intercellular adhesion molecular-1 (ICAM-1), interleukin-13 (IL-13), IL-17 were lower after treatment, with significant difference (P<0.05);there was no significant difference in the incidence of adverse reactions between the two groups.Conclusion The clinical effect of ipratropium bromide in the treatment of asthma and COPD was exactly , ipratropium bromide can significantly improve the pulmonary ventilation function in patients, relieve bronchial spasm, reduce airway resistance, inhibit airway inflammation, and the safety is higher.

Objective To evaluate the values of contrast-enhanced ultrasound in diagnosis of cystic renal cell carcino-ma. Methods A total of 73 patients with renal cystic lesions were included in this study. The image features of ultrasound and contrast-enhanced ultrasound examination were analysed. All of patients underwent surgical treatment and had patholog-ical results. The diagnostic values of the ultrasound and contrast-enhanced ultrasound were analyzed by evaluating the im-age features of cystic renal cell carcinoma. Results There were 64 cases of cystic renal cell carcinoma, 9 cases of benign cyst. With ultrasound and color doppler ultrasound,irregular shape, thickness wall, solid ingredients, divisions and more blood flow signals were found in cystic renal cell carcinoma. Renal cyst showed regular shape, few solid component and thin separation and inconspicuous blood flow signals. In contrast-enhanced ultrasound, cystic renal cancer contrast agent appear-ing time was (15.13±4.21)s, and reached the peak time (23.42±5.68)s, fade time was (28.42±4.27)s. The enhanced mode for fast in and fast out was found in 22 cases (34.3%), fast in and slow out in 30 cases (46.8%), slow in and fast out in 2 cases (3.2%), slow in and slow out in 4 cases (6.4%), and synchronously in and out in 6 cases (9.3%). The hyper-enhancement was found in 42 cases (65.6%), the iso-enhancement and hypo-enhancement in 22 cases (34.4%). In renal cyst, There were three cases out of contrast filling. In other 6 cases, the contrast agent appearing time was (16.67±2.73)s, the peak time was (25.83±3.06)s and fade time was (34.17±4.26)s. The enhanced mode for fast in and fast out was found in 1 case (16.7%), fast in and slow out in 1 case (16.7%) and synchronously in and out in 4 cases (66.6%). The hyper-enhancement was found in 2 cases (33.3%), the iso-enhancement and hypo-enhancement in 4 cases (66.7%). The sensitivity, specificity, positive predic-tive value, negative predictive value and accuracy of ultrasound were 85.9%, 66.7%, 94.8%, 40.0%and 83.6%. The sensitiv-ity, specificity, positive predictive value, negative predictive value and accuracy of contrast-enhanced ultrasound were 92.2%, 77.8%, 96.7%, 58.3%and 90.4%. Conclusion Contrast-enhanced ultrasound can be used in benign and malignan-cy identification of renal cystic lesion.

Objective: To evaluate the effect and safety of neoadjuvant chemotherapy combined with PA-MSHA injection for breast cancer patients. Methods: An open randomized controlled clinical trial was con-ducted. Fourty patients with breast cancer were randomly assigned to neoadjuvant chemotherapy group (the control group, n=20) and neoadjuvant chemotherapy combined with PA-MSHA injection group (the experi-ment group, n=20). The evaluation of therapeutic effect was carried out when the treatment was completed. Kamofsky score was recorded before and after therapy. Venous blood was drawn before and after therapy and immune function (IFN-γ, IL-2, IL-4, and IL-10) and other indicators (Caspase-3, VEGF, MMP-2 and MMP-9) were measured by double antibody ELISA test. Adverse effects of PA-MSHA during therapy were ob-served and recorded. Results: The overall response rate (RR) in the experiment group was significantly higher than that in the control group (P<0.05). No significant difference was found in the pathologic complete remis-sion (pCR) between the experiment group and the control group (P>0.05). In the experimental group, pCR was significantly different before and after therapy (P<0.01). The score in the experimental group was signifi-cantly higher than that in the control group after therapy (P<0.01). With the treatment of chemotherapy and PA-MSHA injection, IFN-γ and IL-2 levels were significantly higher while IL-4 and IL-10 levels were significant-ly lower in the experiment group (P<0.05). A significant increase in serum Caspase-3 and a significant de-crease in serum VEGF, MMP-2 and MMP-9 (P<0.05) after therapy were also observed in the experimental group. The level of serum MMP-9 was decreased significantly (P=<0.05) after therapy in the control group. Con-clusion: Neoadjuvant chemotherapy combined with PA-MSHA injection can significantly improve the RR of breast cancer patients, enhance their cellular immune function, induce the apoptosis and restrain the metasta-sis of breast cancer cells. The PA-MSHA has been proved to be an ideal supplementary therapy for breast cancer.

Objective To investigate the effects and mechanisms of 5-aza-2'-deoxycytidine (5-Aza-CdR) on endometrial cancer cell.Methods In vitro experiments of 5-Aza-CdR were done using human endometrial cancer cell line HEC-1B.Evaluation of cellular proliferation and apoptosis was ascertained respectively using trypan blue exclusion and flow cytometry.RT-PCR and methylation specific PCR(MSP) was done to detect the expression of RASSF1 A mRNA and methylation status of RASSF1 A promoter of HEC-1B cell line.Results (1) The status of cellular growth and apeptosis of HEC-1 B cell line:the growth inhibition effects of 5-Aza-CdR on HEC-1B cell line were both concentration-dependent (P ＜ 0.01) and time-dependent(P ＜0.01),as well as the apoptosis rate of HBC-1-B cell line depended on the dose of 5-Aza-CdR obviously(P ＜0.01).(2)The expression of RASSF1A mRNA of HEC-1B cell line:RASSF1A mRNA was expressed in HEC-1B cell after 5-Aza-CdR treatment,but it was undetectable before the treatment.In the groups with different concentration of 5-Aza-CdR (0.05,0.1,1,5,10 nmol/ml),the expression of RASSF1A mRNA was respectively 0.074±0.004,0.105±0.004,0.167±0.006,0.334±0.005,0.484±0.007,which were remarkably different from the group without 5-Aza-CdR(the expression of RASSF1A mRNA was 0;P ＜ 0.01).(3) The hypermethylation of RASSF1A promoter of HEC-1B cell line:the hypermethylation of RASSF1A promoter was detected in HEC-1B cell line.The status of hypermethylation was decreased after treatment with 5-Aza-CdR of 0.05,0.1,1,5 nmol/ml,meanwhile,both methylation bands and demethylation bands were observed by methylation specific PCR.After the treatment with 5-Aza-CdR of 10 nmol/ml the hypermethylation was absent absolutely.Conclusions (1) In HEC-1B cell line,5-Aza-CdR can inhibit cell proliferation and induce cell apopotosis.(2) 5-Aza-CdR can renew the expression of RASSF1A mRNA of HEC-1B cell line and reverse the hypermethylation of RASSF1A promoter.

Objective:Our aim is to investigate the relationship of VEGF-C,VEGF-D and lymph node metastasis in pancreatic cancer,and to elucidate the role and significance of the cancerous peripheral lymphatic tissue.Methods: The expression of VEGF-C and VEGF-D was assayed by means of immunohistochemistry in 30 pancreatic carcinomas.Results:The positive rates of VEGF-C、VEGF-D were 73% (22/30)、57% (17/30) respectively in pancreatic cancer.The expression of VEGF-C、 VEGF-D in cancerous invasive edge was significantly higher than that in the center of cancerous tissues.There was no correlation between the expression of VEGF-C、VEGF-Dand the site,differentiation,histology types.Ⅲ~Ⅳ stages of pancreatic cancer showed strong expressions of VEGF-C、VEGF-D than that Ⅰ~Ⅱ stages of pancreatic cancer.The positive lymph node in positive VEGF-C and VEGF-D group were higher than that in the negative group.Conclusion:VEGF-C and VEGF-D induced lymphangiogenisis in pancreatic cancer,promoted the tumor cell lymph metastasis.

Renal cell carcinoma is one of the most common tumors in the urinary system. Most of these tumors are malignant. The incidence of renal tumors has increased in recent years. Contrast-enhanced ultrasound examination is increasingly applied in clinics and has become a common diagnostic method for renal lesions. Contrast-enhanced ultrasound can clearly show the organization of tu-mor blood vessels and capillaries and provide dynamic observation of the blood perfusion status of normal human tissues and tumor tis-sues. This review discusses the situation and value of contrast-enhanced ultrasound in renal tumor treatment.

Objective To investigate the relationship between metabolic syndrome (MS) and early recurrence of benign prostatic hyperplasia (BPH) after prostatectomy in the elderly.Methods A total of 152 men aged 65 to 88 years with prostatectomy for more than 5 years were enrolled from August 2008 to March 2013.Blood pressure,body weight,body height,body mass index (BMI) and international prostate symptom score (IPSS) were detected.Levels of fasting blood glucose (FBG),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C),prostate specific antigen (PSA) and maximum urinary flow rate (Qmax) were determined.The correlation of MS-related parameters with prostate volume,IPSS and Qmax were evaluated.Results BPH recurrence rate was significantly higher in patients with MS than without MS (11/26 vs.27/126,x2=12.76,P＜0.001).PSA level,prostate volume and IPSS were significantly higher or larger in the MS group than in the non-MS group [(1.96±0.82)μg/L vs.(1.81±0.90)μg/L,(28.26±5.50) ml vs.(22.38±4.00)ml,(11.12±3.18)vs.(7.11±2.37),F=28.654,44.41,38.56,respectively,P＜0.001],while Qmax value was significantly lower in the MS group than in the non-MS group [(14.77±5.29) ml/s vs.(19.80±4.70) ml/s,F=59.48,P＜0.001].The liner regression analysis showed that in postprostatectomy recurrence group,PV and IPSS had significantly positive correlations with levels of FBG and TG,and BMI (P＜0.05),and had significantly positive correlations with BMI and FBG in non-recurrence group (P＜0.05),while Qmax had a significantly negative correlation with BMI,systolic blood pressure and FBG in the two groups (P＜0.05).Conclusions There is a significant relationship between MS and higher BPH recurrence in elderly patients.

Tumor cells can metabolize glucose through glycolysis to intermediates for biomacromolecule synthesis by inhibiting the activity of the pyruvate dehydrogenase complex (PDC) in mitochondria. In this process, pyruvate dehydrogenase kinases (PDKs) play a key role. The inhibition of the activity of PDKs can effectively block this metabolic pathway, activate mitochondrial oxidative metabolism, and induce tumor cell apoptosis. PDK inhibitors have become a research hotspot in medicinal chemistry, and novel structures targeting classical binding sites have been synthesized. In this paper, recent research progress on PDK inhibitors is reviewed to provide information on these latest entities and to explore their clinical applicability.

A fusion expression vector pPIC3.5K-PDGFR? was constructed to express recombinant receptor tyrosine kinase PDGFR? and the right Pichia pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of strain GS115, a high yield strain named M3 was screened. The strain M3 was cultured in a 5 L fermentor and His-GFP-PDGFR? fusion protein was purified by Ni2+ chelating affinity chromatography. One distinct peak was obtained after elution with 250 mmol/L imidazole. Fusion protein was proved to be 90.08 kD by western blotting, and have tyrosine kinase activity by ELISA. Results showed that the receptor tyrosine kinase PDGFR? was successfully expressed in P. pastoris and could be used as a target for small molecule selective inhibitors screening.

Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion： Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.