[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.

OBJECTIVETo study the effect of acupuncture on the immune function of sepsis patients.

METHODSNinety sepsis patients were assigned to the control group, the thymosin a1 group, and the acupuncture treatment group according to random digit table, 30 patients in each group. Patients in the control group were treated according to the guideline of Surviving Sepsis Campaign (SSC). Patients in the control group received routine treatment. Those in the thymosin alpha1 group additionally received subdermal injection of thymosin alpha1 (1.6 mg), once per day for 6 successive days. Needling at related points such as Zusanli (ST36), Yanglingquan (GB34), Neiguan (PC6), Guanyuan (RN4), and so on, was performed in patients of the acupuncture treatment group, once per day for 6 successive days. T cell subgroups (CD3+, CD4+, CD8+, CD4+ /CD8+) and immunoglobulin levels (IgG, IgA, IgM) were detected. The length of ICU hospital stay, hospital readmission rate, and 28-day mortality were compared among the three groups.

RESULTSAfter six days of treatment, CD3+, CD4+, CD8+, IgG, IgA, IgM, and CD4+ /CD8+ ratio of three groups were all significantly increased (P < 0.01). Of them, CD3+, CD4+, CD8+, IgG, IgA, and IgM increased more significantly in the thymosin alpha1 group and the acupuncture treatment group (P < 0.01). Compared with the control group, the ICU hospitalization length was significantly shortened, the hospital readmission rate and the 28-day mortality were lower in the thymosin alpha1 group and the acupuncture treatment group (P < 0.05, P < 0.01). There was no statistical difference in each index between the thymosin alpha1 group and the acupuncture treatment group (P > 0.05).

CONCLUSIONAcupuncture could adjust the immune function of sepsis patients, improve their immunological indicators and prognoses.

Acupuncture Therapy ; CD4-CD8 Ratio ; Humans ; Length of Stay ; Prognosis ; Sepsis ; diagnosis ; immunology ; therapy ; T-Lymphocyte Subsets ; Thymosin ; analogs & derivatives

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

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

Objective To research the effect and safety of Reteplase for Injection combined with reduced glutathione in the treatment of acute ST segment elevation myocardial infarction.Methods Patients with acute ST segment elevation myocardial infarction (90 cases) in Jianli People's Hospital from February 2015 to February 2016 were selected and divided equally into A,B and C groups according to different treatment methods.Totally 30 patients in group A were given conventional therapy,30 patients in group B were given reduced glutathione for treatment on the basis of conventional treatment,and the other 30 in group C were given combined therapy of reteplase and reduced glutathione on the basis of conventional treatment.The vascular recanalization rate,improvement of effective indicators including creatine kinase isoenzyme (CKMB),troponin Ⅰ (cTnⅠ),left ventricular end diastolic diameter (LVEDd) and left ventricular ejection fraction (LVEF),oxidative stress kinase including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px),and incidence of adverse events of patients were compared among the three groups.Results After thrombolysis,the vascular recanalization rate of group C at different time points (2 h,6 h and 12 h) showed significant difference compared with those of groups A and B (P ＜ 0.05).After the treatment,the effective indicators of the three groups were both significantly improved (P ＜ 0.05),and group C improved more significantly than groups A and B (P ＜ 0.05).After the treatment,the SOD and GSH-Px of groups B and C both significantly improved than group A (P ＜ 0.05),but there was no significant difference between groups B and C.After treatment,the incidence of adverse events of group C was significantly lower than those of groups A and B (P ＜ 0.05).Conclusion Reteplase for Injection combined with reduced glutathione has significant curative effect in the treatment of acute ST segment elevation myocardial infarction,which can effectively improve the cardiac function and inhibition of oxidative stress.It is of higher security but with lower incidence of adverse events.

Objective Autosomal dominant congenital cataract (ADCC) is a common heredit disease.Some known genes and mutated loci related to ADCC have been found.The present study provides other disease-causing genes in the ADCC family.This study was to identify the genetic defect in four generations of a Chinese family with autosomal dominant congenital membranous cataracts and demonstrate the functional analysis of a candidate gene in the family.MethodsThe family with hereditary cataract was recruited from the Tianjin Medical University Eye Center.The family history was collected and recorded.Clinical and ophthalmologic examinations were performed on 6 affected and 14 unaffected family members and periphery blood samples were collected from all of the subjects for genomic DNA preparation.The members were genotyped with microsatellite markers at loci associated with cataracts.Multiplex polymerase chain reaction (PCR) was carried out with microsatellite markers near to candidated loci related to congenital cataracts.PCR products from each DNA sample were separated on a polyarcylamide gel and analyzed.Exclusion analysis was performed by allele sharing analysis and gene sequencing.This trail was approved by the Human Research Ethics Committee of this hospital.The oral informed consent was obtained from all of the subjects before the initiation of this trial.ResultsThe hereditary characteristic of this family was in accordance with the autosomal dominant inheritance with a gene penetrance 100%.Affected members of the family were diagnosed with membranous cataracts without other ocular symptom.The disease-causing gene locus were mapped to 22q11.2-q12.1 at a size of about 2.4 Mbp.The multiple-sequence alignments of complete coding region and splice site of CRYBB1,CRYBB2,CRYBB3,CRYBA4 were obtained but no mutation was found in this study.CRYBB1,CRYBB2,CRYBB3,CRYBA4 were screened by directly sequencing.ConclusionAll known ADCC loci have been excluded from the family.Further study should be carried out to screen other relevant genes or loci in patients with ADCC.The pathogenic gene in the family should be identified through extensive scanning of genes,and a new disease-causing gene may exist in this family.

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.

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.