[Objective]To construct the recombinant adenovirus vector co-expressing human vascular endothelial growth factor 165(VEGF165) and angiogenin-1(Ang-1),and to observe the expression of target gene after transfection.[Method]Molecular biologic techniques were used to clone the genes of VEGF165 and Ang-1,and PCR was used to amplify the DNA sequence of IRES(internal ribozyme entry site) from pIRES2-EGFP.The genes of VEGF165,IRES and Ang-1 were subcloned to pTrack-CMV one by one to get the plasmid named pTrack-CMV-VIA,which contained all the three genes.The linearized shuttle plasmid was cotransformed into BJ5183 bacteria with backbone vector AdEasy-1.Further the recombinant plasmid was packaged and amplified in QBI-293A cells after Pac I digestion to get adenovirus vector pAd-VIA.The expression of the gene of interests was evaluated by fluorescence microscopic analysis of GFP expression and enzyme linked immunosorbent assay(ELISA).[Result]The recombinant adenovirus plasmid was consistent with that shown by sequencing and restriction endonucleases digestion.The recombinant pAd-VIA vector was packaged and amplified successfully in QBI-293A cells.The titer was 2?1010 PFU/ml after amplifying.High positive GFP was expressed in the field through fluorescence microscopy after being transfected by recombinant pAd-VIA.ELISA indicated the expression of the target genes.The difference between the tansfected and non-transfected group was significant(P

Objective To evaluate the serum 25-hydroxyvitamin D [25 (OH)D3] levels in non-diabetic people in Taiyuan community,and its relationship with obesity and abdominal obesity.Methods This crosssectional study enrolled 417 non-diabetic people in Tainyuan community from July to September in 2011 [225 males and 192 females,mean age (47.6 ±7.8) years].For all the enrolled subjects,we collected data about anthropometric indexes,medical history,blood glucose,lipid profile,25 (OH) D3,fasting insulin (Fins) level,homeostasis model assessement for insulin resistance (HOMA-IR),body fat content (％) measured with dualenergy X-ray absorptiometry (DEXA),and intra-abdominal adipose tissue (IAAT) and abdominal subcutaneous adipose tissue (SCAT) calculated based on magnetic resonance imaging at the 4th to 5th lumbar disc level (IAAT≥80 cm2 was considered abdominal obesity).Correlation analysis was performed to assess the relationships of serum 25 (OH) D3 levels with biochemical and body fat indexes.Results The average serum level of 25 (OH) D3 was 44.5 nmol/L in the 417 people,with vitamin D deficiency in 261 cases (62.6％) and vitamin D insufficiency in 109 cases (26.1％).Of the vitamin D deficient and insufficient people,77.6％ had abdominal obesity according to IAAT (IAAT ≥ 80 cm2).According to Pearson analysis result,lg 25 (OH)D3 was negatively correlated with lg Fins,lg HOMA-IR,waist circumference,systolic blood pressure,diastolic blood pressure,total abdominal adipose tissue (TAAT),IAAT (all P ＜0.05).It was also negatively correlated with waist-to-hip-ratio,body mass index (BMI),and body fat content (％),but without statistical significance (all P ＞0.05).In stepwise multiple linear regression model,lg HOMA-IR (t =-4.278,P =0.001) and TAAT (t =-5.146,P =0.002) were independently correlated with lg 25 (OH) D3.Conclusions About 90％ of all non-diabetic population in Taiyuan community have vitamin D deficiency or insufficiency,of whom 4/5 have abdominal obesity.Serum 25 (OH)D3 level is negatively correlated with abdominal adipose tissue,but not significantly associated with BMI.HOMA-IR and TAAT are independent risk factors for vitamin D deficiency.

Objective To develop a new method for describe features of inflow of left ventricle(LV) from color Doppler image by color singular value,and evaluat the value of color singular value in assessment of diastolic dysfunction. Methods Patients with diastolic dysfunction of LV including myocardial infarction (n = 37), angina( n = 35), cardiomyopathy( n = 45) and hypertension ( n =46) were selected and 30 healthy cases as control. CV and AV from color singular value of LV inflow was displayed and calculated automatically by an analyzing system developed in our laboratory. Results Compared with that of control (CV:2.40 0.22 and AV: -0.25 0.05), the absolute value of CV(1.67-1.90 in average)and AV( - 0. 11～-0. 20 in average) from color singular value were decreased significantly( P <0. 001) in groupsof myocardial infarction, angina, cardiomyopathy and hypertension. Conclusions Singular value can be applied to detect color differences in LV inflow from color Doppler signals. Changes of singular value within the diastolic inflow under pathological conditions may reflect the diastolic dysfunction of LV.

PURPOSE: The aim of this study is to investigate the effect of genetic variations and the expression of the reduced folate carrier (RFC) and dihydrofolate reductase (DHFR) on the drug sensitivity to methotrexate (MTX) in different cancer cell lines. MATERIALS AND METHODS: We examined the six human cancer cell lines (MCF-7, AGS, A549, NCI-H23, HCT-116 and Saos-2). The cytotoxicity of MTX was measured by sulforhodamine B (SRB) assay. The expressions of the DHFR and RFC were evaluated by real-time PCR and western blotting. Four single nucleotide polymorphisms (SNPs) of the DHFR and two SNPs of the RFC were genotyped. RESULTS: The IC50s of MTX was in an extensively broad range from 6.05+/-0.81 nM to>1,000 nM in the cell lines. The Saos-2 (>1,000 nM) and MCF-7 (114.31+/-5.34 nM) cells were most resistant to MTX; in contrast, the AGS and HCT-116 cells were highly sensitive to MTX with an IC50 of 6.05+/-0.81 nM and 13.56+/-3.76 nM, respectively. A reciprocal change of the RFC and DHFR mRNA expression was found between the MTX-sensitive AGS and MTX-resistant Saos-2 cells. There was no significant difference in the expression levels of RFC protein in both the AGS and Saos-2 cells, whereas DHFR protein was more increased in the MTX-resistant Saos-2 cells treated with MTX. The genotype of the MTX-sensitive AGS cells were mutant variants of the DHFR; in contrast, the Saos-2 cells had the wild-type of the DHFR. CONCLUSION: In conclusion, this study showed that inverse change of the RFC and DHFR mRNA and protein expression was associated with RFC and DHFR polymorphisms and it is postulated that this phenomenon might play an important role in sensitivity of certain cancers to MTX.
Blotting, Western ; Cell Line ; Folic Acid ; Genetic Variation ; Genotype ; HCT116 Cells ; Humans ; Inhibitory Concentration 50 ; Methotrexate ; Polymorphism, Single Nucleotide ; Real-Time Polymerase Chain Reaction ; Reduced Folate Carrier Protein ; Rhodamines ; RNA, Messenger ; Tetrahydrofolate Dehydrogenase

Objective:To explore any effect of combining intermittent theta-burst stimulation (iTBS) of the cerebellum with physiotherapy on the balance function and gait of stroke survivors.Methods:Thirty-two hemiplegic stroke survivors were divided at random into a treatment group and a control group, each of 16. Both groups received conventional physical therapy. Before their physiotherapy sessions the treatment group received iTBS treatment of the cerebellar hemisphere contralateral to the affected cerebral hemisphere, while the control group was given pseudo-stimulation on the same site. The iTBS was given once a day for 200s each time, 6 times a week for 3 weeks consecutively. Before and after the treatment, as well as 3 weeks later, both groups′ balance was evaluated using the Berg Balance Scale (BBS). Their ability to shift their center of gravity, total length of their shaking trajectory, and maximum shaking diameter were also quantified. Walking ability was assessed using 10m walk test (10MWT) times and the Tinetti Gait Assessment Scale (POMA-G). Lower limb motor function was quantified using the relevant Fugl-Meyer assessment (FMA-LE) and the subjects′ ability in the activities of daily living was measured with the Barthel index (BI).Results:After the 3 weeks of treatment and at the follow-up the average BBS score of the treatment group had improved significantly more than the control group′s average, as had its total track length and maximum shake diameter. The average POMA-G, FMA-LE and BI scores of the treatment group were also significantly better.Conclusions:Combining iTBS with physiotherapy can improve the balance and gait of stroke survivors more effectively than physiotherapy alone.

Guanxin Tongluo capsule is a compound traditional Chinese medicine preparation for the treatment of coronary heart disease. The underlying mechanism may be related to expanding coronary artery, increasing coronary artery blood flow, reducing myocardial oxygen consumption, and decreasing whole blood viscosity, plasma viscosity, and hematocrit. This paper reviews the clinical research progress of Guanxin Tongluo capsule in the treatment of coronary heart disease from the aspects of etiology, pathogenesis, pharmacological research, and clinical research, providing evidence for scientific research and clinical application of Guanxin Tongluo capsule.

PURPOSE: Dose-limiting toxicities of docetaxel are widely considered to be neutropenia, anemia, skin toxicity, and nausea. One of the factors that limit the use of docetaxel is its unpredictability of inter-individual variation in toxicity. MATERIALS AND METHODS: In order to identify the genetic factors that affect the risk of docetaxel-induced toxicities, we recruited patients who received docetaxel chemotherapy. We genotyped 92 patients with single-nucleotide polymorphisms (SNPs) in 5 genes: CYP3A4 (CYP3A4*1B, CYP3A4*18, and CYP3A4*3), CYP3A5 (CYP3A5*2 and CYP3A5*3), ABCB1 (C1236T, G2677G/T, and C3435T), SLCO1B3 (rs11045585), and ABCC2 (rs12762549). RESULTS: Out of 92 patients, 70 had grade 3 or 4 neutropenia; 4 had grade 1 or 2; and 18 had no toxicity (76.1%, 4.3%, and 19.6%, respectively). The findings of the SNP analysis showed that patients with TT genotype of ABCB1 3435C>T polymorphism showed significantly higher risk of neutropenia and anemia (p=0.029 and p=0.044, respectively). There were significant associations between docetaxel-induced leucopenia and 2677G/T of ABCB1 and rs12762549 of ABCC2 (p=0.025 and p=0.028, respectively). In a multivariate analysis, we observed that patients carrying 2677G>T in ABCB1might be associated with higher risk of chemo-resistance when treated with docetaxel (odds ratio [OR], 6.48; confidence interval, 1.92 to 21.94; p=0.003). In a subgroup analysis of non-small cell lung cancer patients, a significant association of tumor response with G2677T/A (OR, 4.54) in ABCB1 and SLCO1B3 (OR, 9.44) was observed. CONCLUSION: Our data suggest that ABCB1 (2677G/T) and SLCO1B3 (rs11055585) might be major genetic predictors of docetaxel-related toxicities in patients receiving docetaxel chemotherapy.
Anemia ; Carcinoma, Non-Small-Cell Lung ; Drug Therapy* ; Genetic Variation* ; Genotype ; Humans ; Multivariate Analysis ; Nausea ; Neutropenia ; Polymorphism, Single Nucleotide ; Skin

The pathological mechanism of allergic rhinitis (AR) involves hierarchically acute and chronic reactions among antigens, immunocytes, immunoglobulins and inflammatory factors. AR, which is characterized by heterogeneity, reacts differently to the current treatment on diverse individuals. Some patients receive unsatisfying clinical outcomes after the treatment. Chinese herbal medicine has become a frequent option to treat AR because it can improve clinical symptoms with excellent safety. The description of Biqiu in ancient Chinese medical literatures is very similar to the clinical manifestation of AR. The major pathogeneses of Biqiu included infection of wind-cold pathogens, deficiency and cold in the lung, and evil depression into heat and wind. Therefore, in modern traditional Chinese medicine （TCM）, Biqiu is considered as an integration of AR, allergic sinusitis and vasomotor rhinitis. According to the pathogenesis and clinical manifestations of Biqiu (including AR), this disease belongs to wind disease in TCM. But modern TCM focuses on deficiency of internal organs and ignores wind evil in syndrome differentiation, contrary to the fact that wind medicine is always used in treating AR. This even leads to an abnormal phenomenon that clinical studies attach little importance to syndrome differentiation. In this paper, we started from the phenomenon that wind medicine is always used in TCM for AR, considering TCM literatures and the study results of modern therapy for wind diseases, and put forward the ideas as follows: AR belongs to exogenous wind disease in TCM, and wind pathogen is the major pathogen, in clinical syndrome differentiation, we should attach more importance to wind evil, and dispelling wind and eliminating dampness is the key to treatment. Meanwhile, based on the fact that most of the classical prescriptions with good effectiveness for AR originated from Treatise on Febrile Disease, mainly including the wind medicines for treating exogenous diseases, it can be inferred that the wind-dispelling and itch-arresting effects of wind medicines are helpful to alleviate or block the allergic reaction of nasal mucus and achieve the goal of relieving itching and sneezing, and the wind-dispelling and dampness-eliminating effects are helpful to alleviate the edema of nasal mucus and achieve the goal of relieving running nose and nasal obstruction. In modern researches, it is also found that classical prescriptions which take wind medicine as a principle component and wind medicine used in AR treatment have good anti-inflammatory, anti-allergic and immune function adjusting effects. This indicates that dispelling wind and eliminating dampness and the wind medicine treatment under its instruction not only correspond to TCM theory, but also have modern science evidences.