[Objective]To investigate the effect of transforming growth factor ?1(TGF-?1) genes transfection on the proliferation and differentiation of chondrocyte by adenovirus vector,and to observe the quality in repair cartilage defect through constructing tissue-engineered cartilage.[Method]Using replicat ion defective adenovirus Adeno-X~(TM) as a carrier,Articular chondrocytes were transferred by high titers level recombinant adenovirus taking TGF-?1 genes.Ultromicrostructure,prolife ration anddifferentiation were observed by light and electron microscope,flow cytometry and so on.Induce expression of exogenous genes coding protein and cartilage cell phenotype were detected by Immunocytochemistry and Northern blot.Gross observation,histology and effect classification were used to evaluate results in repair cartilage defect by combi nation of infected chondrocytes and bone matrix gelatin(BMG).[Result]Immunocytochemistric staining showedthe expression of exogenous gene coding proteins after infection,and procollagen Ⅱ mRNA in cells was detected increasingly by Northern blot.TGF-?1 stimulated proliferation of chondrocytes in primary cultured,and resulted in recruitment of articular chondrocytes into S-G2/M phase.Chondrocytes seeded onto BMG showed high level of proliferation.Transplantation in vivo showed: cartilage defect was repaired satisfactorily by Combination of BMG and infected chondrocytes,and the structure of repair tissue got close to normal articular cartilage.[Conclusion] Articular chondrocytes prolifera tion could be stimulated by adenovirus vector taking TGF-?1 genes,which could be used to constructed tissue-engineered cartilage,and applied in repairing joint cartilage defect.

Objective To observe the impact of the five-emotion restrictive therapy on coronary heart disease. Methods109 hospitalized patients with coronary heart disease were randomly divided into a experimental of 56 patients and a control group of 53 patients according to length of stay. The control group was treated with the conventional therapy; the experimental group was imposed five-emotion restrictive psychological intervention on the basis of conventional therapy. The investigation was done using self-rating anxiety scale(SAS) and self-rating depression scale(SDS) on 1 day after admission and ld before discharge respectively. SAS and SDS scores were surveyed. At the same time Holter monitoring and normal ECG test were compared. Results Compared with control group, the experimental group 1 d before discharge of the SAS and SDS scores were (41.9±7.4) and (38.6±8.4), and the difference was statistically significant (P＜0.01) . The total effective rate of Holter monitoring and normal ECG was 64.15％ and 92.85％ in the control and experimental group, and the difference was significant (P＜0.01) . ConclusionThe pathological seven emotions have a certain affection on the incidence of coronary heart disease, treatment and prognosis of the course. Five-emotion restrictive psychological intervention can adjust timely psychological state of patients, eliminate negative emotions, and significantly help the incidence, treatment and prognosis of the disease.

Objective To study the effect of dialysis adcquacy,microinflammation and residual renal function on nutritional status of hemodialysis patients.Methods One hundred and fourteen patients were enrolled in this study.Kt/V,?_2-MG and serum iPTH were measured as markers of hemodialysis adequacy.Nutritional evaluation included MQSGA,Alb,Hb,TF,IGF-1,IGFBP-3 and anthropometrics such as HGS,BSF,TSF,MAC,MAMC and AMA.Serum IL-6,TNF-?and CRP were detected to assess microinflammation.Urinary volume of 24 hours was measured to investigate the residual renal function (RRF).Results (1)There were different correlations and regressive associations of Kt/V,iPTH and?_2-MG with HGS,MAMC,AMA,Alb,Hb,nPCR,IGF-1 and MQSGA respectively.(2) There were significant correlations and regressive associations of RRF to HGS,TSF,MAMC,Alb,nPCR and IGF-1 within the first year of hemodialysis.(3) There were different correlations and regression relationships of IL-6,TNF-?and CRP with HGS、MAMC、AMA、Alb、TSF、Hb、nPCR、IGF-1 respectively.(4) Multivariate analysis showed that Kt/V,iPTH,IL-6, TNF-?,?_2-MG and RRF were influencing factors,among them,Kt/V,iPTH,IL-6 and TNF-?were independent predictors of nutritional status.Conclusions Hemodialysis adequacy and micruinflammation may impact on nutritional status.Residual renal function may be involved in nutritional status in the first year of hemodialysis.Kt/V,iPTH,IL-6 and TNF-?are independent factors affecting nutritional status.

Objective To Discuss the impacts of different dosage of atorvastatirs on serum hsCRP,IL-10 and MCP-1 levels on post-intervention patients with coronary stenting. Methods 93 post-intervention patients with coronary stenting were selected and randomly divided into 3 groups.Each group took different dosage of oral atorvastatins after the operation for more than one week.The dosage for each group was 10 mg,20 mg and 40 mg,respectively.Each patient was phlebotomized for three times,which are 24 hours before the operation,24 hours after the operation and one week after the operation.Serum MCP-1,IL-10 and hs-CRP levels were measured by enzyme linked immunosorbent assay(ELISA)and immunoturbidimetry(ITM). Results Serum hs-CRP and MCP-1 levels of post-intervention patients were significantly higher than those of pre-intervention.This illustrated that the serum hsCRP and MCP-1 levels were closely related to PCI.Serum hs-CRP and MCP-1 levels decreased in those patients one week after operation which proves they are negatively correlated with the dosage of atorvastatins.There was no statistic evidence to prove the correlation between different dosage of atorvastatins and the level of serum IL-10.The ratio of MCP-1/IL-10 at 24h post-intervention patient was significantly higher than pre-intervention,which proves the ratio was negatively correlated with the dosage of atorvastatins. Conclusion Atorvastatins decreases serum MCP-1 and hs-CRP levels after PCI.Serum MCP-1 and hs-CRP levels were negatively correlated with the dosage of atorvastatins.

Objective To investigate the serum homocysteine (HCY)levels of patients with various liver diseases,and discuss the clinical value of combined detection of HCY and other liver function indicators in liver disease diagnosis.Methods Se-lected 123 cases of inpatients with different liver diseases (79 cases of male,44 cases of female),including 39 cases of chronic active hepatitis,52 cases of cirrhosis and 32 cases of primary hepatic cancer from Department of Gastroenterology,and 90 ca-ses of healthy persons as control group from Out-patient Health Examination Center of Shaanxi Provincial People’s Hospi-tal.Detected serum HCY and some indicators of liver function (TBIL,ALT,GGT and TBA)in different liver disease groups and normal control group with Roche MODUALR automatic biochemical analyzer,and did statistic analysis.Results The serum HCY levels of chronic active hepatitis,liver cirrhosis and primary hepatic cancer were 16.25± 3.98μmol/L,20.89± 4.26μmol/L and 22.92 ± 6.08μmol/L respectively,and significantly higher than those in normal control group (8.16 ± 4.03μmol/L,t=5.352~9.021,P<0.01).The serum HCY levels of liver cirrhosis and primary hepatic cancer group were higher than that of chronic active hepatitis group (P<0.05).Serum levels of HCY were obviously lower after treatment than that before treatment (P<0.05)for three groups of liver diseases.The positive rate of HCY was higher than those of other liver function indicators (P<0.05).Conclusion Serum homocysteine may be as one of the most important indexes of liver damage,and the combined detection of other liver function indicators may have important clinical value for liver disease diagnosis,differential diagnosis,treatment and prognosis.

OBJECTIVETo observe the clinical effect of acupuncture on acute heart failure (AHF) patients.

METHODSTotally 60 patients who were diagnosed as AHF were assigned to the acupuncture group and the control group, 30 in each group. Those in the control group received inotropic agents, preload and afterload reducing therapy, anti-infection and so on. Besides, those in the acupuncture group received needling at relative points, once daily for 5 consecutive days. The mean arterial pressure (MAP), heart rate (HR), cardiac index (CI), stroke volume index (SI), left ventricle working index (LCWI) were monitored by thermodilution pulse-indicated continuous cardiac output (PiCCO) technique. Changes of the aforesaid data were compared between before and after treatment. The ICU length of stay, readmission rate,and the 28-day mortality were also compared between the two groups.

RESULTSAfter 5 days of the treatment, CI, SI, and LCWI increased more obviously (P < 0.01), HR and MAP decreased significantly (P < 0.05). Of them, CI, SI, and LCWI increased more obviously in the acupuncture group (P < 0.05).There was no obvious difference in HR or MAP between the two groups (P > 0.05). Compared with the first day of admission in the same group, CI, SI, and LCWI obviously increased in the acupuncture group from the second day (P < 0.05). HR started to decrease since the fourth day (P < 0.05), and MAP began to decrease until the fifth day (P < 0.05). CI, SI, and LCWI started to increase in the control group from the third day (P < 0.05); HR and MAP both began to decrease since the fifth day (P < 0.05). Compared with the control group, the ICU length of stay was obviously shortened in the acupuncture group (P < 0.05). The readmission rate and the 28-day mortality rate were lower than those of the control group (P < 0.05).

CONCLUSIONThe combination of acupuncture and Western medical therapy might strengthen acute heart failure patients' heart functions, elevate the therapeutic effect, and improve the prognosis.

Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Female ; Heart Failure ; physiopathology ; therapy ; Heart Rate ; Humans ; Male ; Middle Aged ; Stroke Volume

Animals ; Child, Preschool ; Diagnosis, Differential ; Humans ; Male ; Toxoplasma ; Toxoplasmosis ; diagnosis ; drug therapy ; parasitology

Objective To solve the problem of castration resistant prostate cancer (castrationresistant prostate cancer,CRPC) the problem of drug resistance by studing the expression of the CUDC-101 inhibitor of castration resistant prostate cancer androgen receptor splice variant 7.Methods In this study,the expression of AR-V7 protein in prostate cancer cell lines PC-3,VCaP,22Rv1,LNCap was detected by imnmunoblotting between April 2015 and April 2016,and the highest expression level of cell lines was selected follow-up experiments.Through the cell proliferation and activity experiments,the epigenetic inhibitors:histone deacetylase inhibitor CUDC-101,histone methylation inhibitor DZNeP,DNA methylation inhibitor gemcitabine,histone acetyltransferase inhibitor MG149 to select an inhibitor that reduces the expression of AR-V7 protein in CRPC cells.22Rv1 cells were treated with 30 nmol and 300 nmol of CUDC101 and 1,10 and 20 μmol of Enzalutamide (MDV3100),respectively,and their inhibitory effects on the growth of 22Rv1 cells were examined.Results The results of immunoblotting showed that AR-V7 protein was only expressed in CRPC cell line 22Rv1 and negative in non-CRPC cell line.The expression of AR-V7 in 22Rv1 cells treated with CUDC-101 was significantly lower than that of negative control.While other inhibitors had no effect on the expression of AR-V7.In the cell proliferation and activity assay,the inhibitory rates of 30 nmol CUDC-101 and 1,10 and 20 μmol MDV3100 were 10％,35％ and 45％,respectively,higher than that of MDV3100 alone.28％ and 42％,the difference was statistically significant (P ＜ 0.05).The inhibitory rates of 300 nmol CUDC-101 and MDV3100 were 30％,60％ and 65％,respectively,which were significantly higher than those of MDV3100 alone (P ＜ 0.05).Conclusions CUDC-101 can inhibit the castration resistant prostate cancer androgen receptor splice variant 7 expression,and solved the resistance problem of CRPC.

A study on the microbial transformation of glycyrrhetinic acid (GA) was conducted by a fungus, Cunninghamella blakesleeana CGMCC 3.970 systematically. After incubation with the cell cultures of C. blakesleeana CGMCC 3.970 at 25 degrees C for 7 days on a rotary shaker operating at 135 r x min(-1), GA was converted into one major product and five minor products. The products were extracted and purified by solvent extraction, macroporous adsorbent resin, silica gel column chromatography, and semi-preparative RP-HPLC chromatography. Their structures were identified as 3-oxo-15α-hydroxy-18β-glycyrrhetinic acid(1), 3-oxo-15β-hydroxy-18β-glycyrrhetinic acid (2), 7β,15α-dihydroxy-18β-glycyrrhetinic acid (3), 3-oxo-7β, 15α-dihydroxy-18β-glycyrrhetinic acid (4), 7β-hydroxy-18β-glycyrrhetinic acid(5) and 15α-hydroxy-18β-glycyrrhetinic acid(6) by the analyses of MS, 1H-NMR and 13C-NMR spectroscopic data respectively. Among them, 2 was a new compound. These results suggest that C. blakesleeana CGMCC 3.970 has the capability of selective ketonization and hydroxylation for GA. [Key words] glycyrrhetinic acid; Cunninghamella blakesleeana CGMCC 3. 970; microbial transformation
Biotransformation ; Cunninghamella ; metabolism ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; metabolism ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Aim To construct a eukaryotic expression vector of human MCHR2 and to analyze the stable expression and signal transduction pathways of MCHR2 in CHO cells. Methods The full-length MCHR2 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and inserted into eukaryotic expres-sion vector pcDNA3.1(+), resulted in the recombinant expression vector pcDNA3.1-MCHR2. The recombinant plasmid was confirmed by the restriction enzymatic digestion and DNA sequencing analysis, and then transfected into CHO cells by lipofectamine. A stably-transfected cell line was obtained by the dominant G418 selection, and the expression of the MCHR2 gene on transcription and translation levels were identified by RT-PCR and Western blot. Signal transduction pathways mediated by MCHR2 were analyzed by measurement of intracellular cAMP and calcium. Results The eukaryotic expression vector pcDNA3.1-MCHR2 was successfully constructed and the MCHR2 gene was stably transfected into CHO cells. A stably-transfected cell line was established and the MCHR2 gene was efficiently transcribed and translated. MCH stimulation had no effect on the production of cAMP, however, it could induce a clear and transient increase of intracellular calcium, suggesting that MCHR2 was only coupled to Gq protein. Conclusion The stable expression of MCHR2 and analysis of its signal transduction pathways in CHO cell line provided a solid experimental foundation for further studies on the function of the MCHR2 gene in vitro.