Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; GATA6 Transcription Factor ; genetics ; Gene Expression Regulation ; Muscle, Smooth, Vascular ; drug effects ; Myocytes, Smooth Muscle ; drug effects ; Phenotype ; Rats ; Rats, Sprague-Dawley ; Simvastatin ; pharmacology

Objective To investigate the effect of GATA-6 on endogenous carbon monoxide(CO) inhibited pulmonary vascular smooth muscle cells(PVSMCs) proliferation induced by platelet-derived growth factor(PDGF).Methods Tissue mass culture was done to get PVSMCs artery in SD rats.The PVSMCs were stimulated to proliferation by PDGF(20 ?g/L),and the 3 different concentrations of hemin[a substrate and inducer of heme oxygenase-1(HO-1)] add into the cultures to induce CO production.The PVSMC cell proli-feration was detected by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method,DNA synthesis was detected by thymidine incorporation,and GATA-6 mRNA expression was detected by reverse transcription polymerase chain reaction(RT-PCR).Results CO inhibited the PVSMCs proliferation induced by PDGF in a dose-dependent and time-dependent manner.Hemin with high concentration markedly inhibited the proliferation and DNA synthesis of PVSMCs.After 2 hours with PDGF,the expression of(GATA-6) mRNA markedly down-regulated,and began returned after 6 hours.However,CO could reversed this down-regulation.Conclusions CO can inhibit the proliferation of PVSMCs induced by PDGF.PDGF can result in the expression of GATA-6 mRNA down-regulated;the down-regulation is reversed by CO.This study suggested that CO maybe inhibit PVSMCs proliferation by regulating expression of GATA-6.

This research is to establish an HPLC method for the simultaneous determination of ophiopogonin D, ophiopogonin D', ophiopogonin C, deacetylophiopojaponin A and ophiogenin-3-O-α-L-rhamnosyl-(1-->2)-β-D-glucoside in Ophiopogonis Radix. HPLC-ELSD analysis was performed on a Kromasil 100-5 C₁₈ column (4.6 mm x 250 mm, 5 µm), with the mobile phase of acetonitrile (A) -water (B) in gradient elution mode (0-45 min, 35%-55% A), at a flow rate of 1 mL · min⁻¹. The column temperature was 35 °C and the drift tube temperature was 100 °C in a gas flow rate of 3.0 L · min⁻¹. The result showed that baseline of all the 5 constituents was well separated, and every constituent had wide linearity range and good linear relation (r > 0.999). The recovery rate was between 95.75% and 103.1%. The new established method for simultaneous determination of saponin constituents in Ophiopogonis Radix was sensitive and has good, repeatability. It could be applied to quality evaluation of Ophiopogonis Radix.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification

Objective To investigate the correlation between serum levels of homocystein (Hcy),Cystatin C (Cys C)and cognitive impairment in patients with chronic kidney disease (CKD).Methods 78 patients with CKD from January 2016 to May 2017 in Hanzhong People's Hospital in Shaanxi were enrolled for the study and divided into stage 3 groups (n 27),stage 4 group (n=28) and stage 5 group (n =23) according to stage of CKD.Montreal cognitive assessment scale (MoCA) was performed to evaluate the cognitive level of patients,and they were divided into cognitive impairment group and normal group.Serum levels of Hcy,Cys C,creatinine (SCr) and urea nitrogen (BUN) were compared among groups,and the correlation between them and cognitive level was analyzed.Results ①Serum levels of Hcy,Cys C,SCr and BUN in stage 3 group were significantly lower than that in stage 4 and 5 group,and these indexes in stage 4 group were significantly lower than that in stage 5 group (F=19.221～66.539,all P＜0.05).The score of MoCA in stage 3 group was significantly lower than that in stage 5 group,and the incidence of cognitive impairment was significantly higher than that in stage 5 group (F/x2 =3.993～6.358,all P＜0.05).②The serum levels of Hcy,Cys C,SCr and BUN in cognitive impairment group were significantly higher than in normal group (t =2.198 ～ 3.835,all P＜ 0.05).③ Pearson analysis result showed that MoCA score was negatively correlated with serum Hcy,Cys C and SCr (r=-0.370,-0.392,-0.253,P＜0.05),but there was no sig nificant correlation between score and BUN (r=-0.211,P＞0.05).④For prediction of cognitive impairment,area under the receiver operating curve of serum levels of Hcy,Cys C,SCr and BUN were 0.818,0853,0.751 and 0.709 (P＜0.05).Conclusion Serum levels of Hcy,Cys C were significantly negatively correlated with cognitive function in patients with CKD,whose predictive value were higher than that ot traditional renal function indexes such as SCr and BUN.

Objective To investigate the effects of radix salviae militiorrhizae (RSM) on acute lung injury induced by"two hits"and to study its probable mechanism.Method Thirty Wister rats were randomly divided into three groups:namely normal control group,model group and RSM treatment group.The model was created by"two-hits"in which 0.2 ml/kg oleic acid was injected into tail vein first,and then 2 mg/kg lipopolysaccharide was administered four hours later.After model rats sacrificed,the pathological changes of lung were observed,and lung wet/dry weight ratio,protein content,and the ratio of neutrophiles in brochoalveolar lavage fluid (BALF) were calculated.In addition,the expression of Fas,FasL protein and apoptosis were evaluated by immunohistochemical studies and TUNEL technique.Results The acute lung injury rat model was successfully induced by"two hits".The gross and micrographic injury of lung was milder in RSM treatment rats than in model rats.The W/D ratio,protein contents and the ratio of neutrophiles in BALF were also markedly reduced in comparison with model rats,while the expression of Fas and Fasl,and the apoptosis index in model rats were significantly increased compared with other two groups.Furthermore,it showed a positive correlation between the expression of Fas,FasL,and the number of cell with apoptosis.Conclusions RSM shows a protective effect on ALT rats caused by"two hits"likely reaulted from inhibiting the expressions of Fas and Fasl,which are associated with the cell apoptosis of lung tissue.

Photosynthetic bacteria(PSB) showed great promise in biohydrogen production. Chromatium vinosum was able to utilize the fermentation waste of Klebsiella oxytoca for both photo-fermentative and dark-fermentative hydrogen production. The content of residual sugars and main organic acids decreased obviously after hydrogen production by C.vinosum. The maximal hydrogen production of C.vinosum was obtained at pH 6.5 adding extra 0.1%(W/W) NH_4Cl. Under photo-fermentative conditions, the content of butyric acid decreased by 54.38%, and the maximal hydrogen yield was 36.97 mL/mg cell. Under dark-fermentative conditions, the content of butyric acid decreased by 36.1% and the maximal hydrogen production was achieved as 37.50 mL/mg cell.

Cognitive impairment is a common complication of diabetes. Hippocampus plays an important role in cognitive function. In hyperglycemia, synaptophysin, a crucial synaptic vesicle membrane protein in hippocampus neuron is found to be down-regulated. Recent evidences have shown that angiotensin IV can facilitate memory acquisition and recovery. However, whether it can also improve cognitive functions of diabetic rats with cognitive disorder, and the possible mechanisms are uncertain. Hence, the objectives of this study. Forty fi ve Sprague Dawley male rats were randomly divided into three groups: Control, diabetic group and diabetes with angiotensin IV treatment group. The cognitive functions, mainly learning and memory of the rats were evaluated using Morris water maze task. The synapses ultrastructure, relative mRNA concentrations and protein expression levels of synaptophysin in hippocampus CA1 area were estimated using transmission electron microscope, RT-PCR, immunohistochemistry and western blotting, respectively. Our study showed that in the diabetic rats with angiotensin IV treatment, the cognitive impairment as measured by Morris water maze task improved, the ultrastructure of synapses in hippocampus reversed, the relative mRNA concentrations and protein levels of synaptophysin in hippocampus signifi cantly increased, when compared with diabetic rats. We conclude that angiotensin IV plays an important role in improving cognitive function of diabetic rats. The possible mechanisms are up-regulating the expression of synaptophysin and normalizing the ultrastructure of synapses in hippocampus.

Objective To evaluate the efficacy and safety of recombinant adenovirus-p53 (rhAd-p53) combined with neoadjuvant chemotherapy in treatment of locally advanced cervical cancer.Methods Forty patients with stage Ⅰ R2-Ⅲ A locally advanced cervical cancer were randomly divided into 2 groups,gene therapy + neoadjuvant chemotherapy group (rhAd-p53+PVB group,n =20.They received one course of chemotherapy consisting of PVB.rhAd-p53 solution 1 ×1012 VP was injected intratumorally every three days for three circles since the 3rd day of PVB chemotherapy) and chemotherapy group (PVB group,n=20,the above course of chemotherapy was conducted).The volums of tumors was observed.Patients were monitored for adverse event.The expression of VEGF,p53 pertein and MVD in tumor tissue was detected by immunohistochemistry.Results The evaluation was performed three weeks after the completion of chemotherapy.The PVB group response rate (CR+PR) was 75 ％,while the effective rate was 95 ％ of the PVB combined with gene group.After using of the PVB chemotherapy,the tumor was shrunk by (11.42±2.78) cm2.However,the volums of tumor were significantly shrunk by (15.25±4.00) cm2 using the PVB combined with gene therapy,and P ＜ 0.05.The positive expression rate of VEGF,p53 protein and MVD were reduced respectively in PVB group and rhAd-p53 + PVB group with statistic significance.There were no additional adverse events by recombinant adenovirus-p53 combined with neoadjuvant chemotherapy.Conclusion A potentially gene therapeutic agent for cervical cancer treatment,intratumoral injection of rhAd-p53 is effective.

A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis
Alkyl and Aryl Transferases ; biosynthesis ; genetics ; Azulenes ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Open Reading Frames ; Polyisoprenyl Phosphates ; Recombinant Proteins ; biosynthesis ; Sesquiterpenes ; metabolism ; Sesquiterpenes, Guaiane ; Thymelaeaceae ; enzymology ; genetics

Ataxia Telangiectasia Mutated Proteins ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Cell Cycle Proteins ; metabolism ; Checkpoint Kinase 2 ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Protein-Serine-Threonine Kinases ; metabolism ; Tumor Burden ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism