For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; Hepatic Stellate Cells ; High-Throughput Screening Assays ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

Objective To assess the surgical effectiveness of pelvic endometriosis (EMS) by Laparoscopic. Methods Retrospective analysis was performed in 1272 pelvic EMS patients underwent laparoscopic surgical treatment. All patients were classified into four groups according to the R-AFS,63 patients of Ⅰ stage,44 patients of Ⅱ stage,475 patients of Ⅲ stage and 690 patients of Ⅳ stage. Symptoms of EMS,surgical effect and follow-up outcomes were compared. Results In all stages, there were no significant differences on moderate to severe dysmenorrhea,increase in CA125 and infertility (Ps ＞ 0. 05 ). In the EMS patients combined with AM or/and DIE, the rates of the moderate to severe dysmenorrhea, CA125 increase and infertility were 84.6% (11/13 ) ,92.3 %(12/13) and 53.8% (7/13)in stage Ⅰ ,81.8% (9/11) ,90. 9% ( 10/11 )and 54.5% (6/11) in stage Ⅱ ,84.8%(173/204) ,93. 1% (190/204) and 47.1% (96/204) in stage Ⅲ ,and 81.6% (213/261) ,91.2% (238/261) and 46.4% ( 121/261 ) in stage Ⅳ, respectively. The rates in non-consolidated AM or/and DIE patients were 46.0%(23/50),62.0% (31/50) and 22.0% ( 1 1/50) in stage Ⅰ ,45.5 % ( 15/33 ),54.5% ( 18/33 ) and 18.2% (6/33)in stage Ⅱ ,41.7% ( 113/271 ) ,62.7% ( 165/271 ) and 23.6% (64/271) in stage Ⅲ ,and 47.3% (203/429),60.1% (248/429) and 21.7% (93/429). The difference between the two groups with and without consolidated AM or/and DIE had statistical significance( P ＜ 0.05 or P ＜ 0.01 ). After the treatment, the scores of life's energy, ache,emotion,sleep,social activity and body movement of NHP were significantly superior than those before treatment in all patients ( P ＜ 0.01 ). The surgery time-consuming of Ⅲ and Ⅳ stage patients were (64.5 ± 18.4) min and (61.7 ± 17.1 ) min respectively, which were significantly higher than that of Ⅰ and Ⅱ stage ( ( 31.9 ± 12.3 ) min and (40.3 ± 10. 6) main ] ( t = 20.25, P ＜ 0.01 ). The massive hemorrhages and the internal damage organs occurred in Ⅲ and Ⅳ stage surgery [2.9% and 3.5% ,respectively] ,whereas much less in Ⅰ and Ⅱ stage (0.8% and 1.4% ) ,with no significant difference(P ＞0. 05). After treatment,the rate of symptoms recurrence of Ⅲ and Ⅳ stage patients respectively were 21.4% ,which were higher than that of Ⅰ and Ⅱ stage patients ( 2.3% ) (P ＜ 0.05 )The rate of pregnancy in Ⅲ and Ⅳ stage patients ( 15.4% ) were lower than that of Ⅰ and Ⅱ stage patients (69. 6% ) ( P ＜ 0. 01 ). Conclusions Pelvic EMS decreased the quality of life. EMS patients combined with adenomyosis or/and deep infiltrating EMS have more severe pelvic pain symptoms, less surgical effectiveness and more serious complications. Ⅲ and Ⅳ stage patients are more liable to symptoms recurrence and lower pregnancy rate.

Objective To construct the new diet intervention scheme of dyslipidemia of university teachers, discuss the effect of this plan on eating behavior change. Methods Choose two groups of college teachers in Harbin, who were hyperlipemia and had physical examinations from June to September, 2014. 44 teachers from one college were conducted the new dietary intervention as experimental group, 37 teachers from another college were the control group, used the usual method. Compared two groups before and after the intervention of dietary behavior and blood lipid. Results The experimental group′s dietary behaviors changed strongly after implement the new dietary intervention, the scores of DTS before the experiment (58.82 ± 18.47) points,3 months after the experiment (48.36 ± 14.25) points and 6 months after the experiment (44.18±14.92) points were statistically significant (F=21.308, P < 0.01). There was no statistically significant difference in control group (F = 1.129, P > 0.05), respectively (60.51 ± 16.91) points, (57.19 ± 16.35) points, (56.92 ± 21.35) points. After 6 months, the experimental group′s subjects of TC was (4.28±3.73) mmol/L, the control group was (6.23±1.04) mmol/L, the difference was statistically significant (t = 3.082, P < 0.05), there were no significant differences in TG, LDL-C and HDL-C of the two groups (P>0.05). Conclusions The new dietary guidance plan can effectively improve and maintain the dietary behavior in hyperlipidemia college teachers, and decrease the blood lipid level.

In our study, 198 types of pesticides in 120 types 333 lots of traditional Chinese medicine (TCM), which were reasonably classified according to its matrix property, were determined by using the pretreatment platform and gas chromatography-mass spectrometry method. As a result, 158 were contaminated with pesticides. However, the content of pesticides in most TCM was very low. In addition, types of pesticides were different in different part of materia medica. In conclusion, the current status of pesticide residues pollutants in TCM was summarized, and the result can provide proof for the formulation of maximum residue limit. The new species of herbs and the new detecting index should be electively monitored in Chinese Pharmacopeia.
China ; Drug Contamination ; prevention & control ; Pesticide Residues ; analysis ; Plants, Medicinal ; chemistry ; Quality Control

Objective To investigate the impact of miR-200c overexpression on colon cancer cell proliferation ability and the related mechanism.Methods MicroRNAs which may combined with the transcription factor AP-2α were screened and forecasted by the bioinformatics database,while its eukaryotic expression plasmids and specific inhibitor were synthesized.Plasmids PEZX-miR-200c,PEZX-NC,pmirGLO-AP-2α3'UTR,pmir-GLO and the specific inhibitors miR-67-inhibtor,miR-200c-inhibitor were transfected in vitro into colon cancer HCT-116 and SW480 cells and the HEK293T cell by Lipofectamine2000.The expression of AP-2α mRNA and protein in colon cancer cells was analyzed by qRT-PCR,Westem blot and immunocytochemical staining.CCK-8 assay and flow cytometry were adopted to observe the effect of miR-200c on colon cancer cells proliferation and apoptosis.Dual-Luciferase assay experiments were performed to observe the relative luciferase activity induced by miR-200c.Results The proliferation activity was significantly decreased in anti-miR-200c/SW480 group,while in PEZX-miR-200c/HCT-116 group,it was higher than that in PEZX-NC/HCT-116 group.The apoptosis ability was significantly increased in anti-miR-200c/SW480 group [(78±0.7) ％ vs (66±1.1) ％,P ＜ 0.05].The expression of AP-2o both in mRNA and protein levels was decreased in PEZX-miR-200c/HCT-116 group,while the protein level was increased in Anti-miR-200c/SW480 group.The relative luciferase activity inhibited by miR-200c was decreased in HEK-293T cells transfected with PEZX-miR-200c and pmir-GLO-AP-2α3' UTR (0.51±0.09 vs 0.98±0.04,P ＜ 0.01).Conclusion MicroRNA-200c could promote cell proliferation ability by targeting transcriptional factor AP-2α in human colorectal cancer cells.

Objective To investigate the association between methylthioadenosine phosphorylase (MTAP) gene single nucleotide polymorphisms (SNP) and myocardial infarction (MI) in the Chinese Han ethnicity. Methods 432 patients suffered from myocardial infarction and 430 controls were involved for case and control groups, respectively. Nine tag SNPs in MTAP gene were selected and genotyped. Results We found no significant association of selected tag SNPs with MI in all of the samples. However, in stratified analysis, significant association was observed at rs7027989 in male subjects. The risk of MI increased by 26% (P=0.005) for male subjects of minor allele carriers in a dominant model. The increased risk of MI at rs7027989 remained significant after adjusting for confounding factors. Conclusion MTAP gene might be involved in the etiology of MI in Chinese Han ethnicity.

The samples of sulfur-fumigated Paeoniae Alba Radix acquired both by random spot check from domestic market and self-production by the research group in the laboratory were used to evaluate the effects of sulphur fumigation on the quality of Paeoniae Alba Radix by comparing sulfur-fumigated degree and character, the content of paeoniflorin and paeoniflorin sulfurous acid ester, and changes of the fingerprint. We used methods in Chinese Pharmacopeia to evaluate the character of sulfur-fumigated Paeoniae Alba Radix and determinate the content of aulfur-fumigated paeoniflorin. LC-MS method was used to analyze paeoniflorin-converted products. HPLC fingerprint methods were established to evaluate the differences on quality by similarity. Results showed that fumigated Paeoniae Alba Radix became white and its unique fragrance disappeared, along with the production of pungent sour gas. It also had a significant effect on paeoniflorin content. As sulfur smoked degree aggravated, paeoniflorin content decreased subsequently, some of which turned into paeoniflorin sulfurous acid ester, and this change was not reversible. Fingerprint also showed obvious changes. Obviously, sulfur fumigation had severe influence on the quality of Paeoniae Alba Radix, but we can control the quality of the Paeoniae Alba Radix by testing the paeoniflorin sulfurous acid ester content.
Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Fumigation ; methods ; Paeonia ; chemistry ; Quality Control ; Sulfur ; chemistry

Background 5-hydroxytryptamine 2A receptor (5-HT2A) gene has been regarded as a candidate gene for susceptibility to schizophrenia. In particular, the 5-HT2A receptor has received much attention because it demonstrates to be an important site of action of atypical antipsychotic agents to alleviate negative symptoms. The current study investigated whether the 5-hydroxytryptamine 2A receptor (5-HT2A) gene T102C polymorphism was associated with treatment response to risperidone in the first episode Chinese patients with schizophrenia.Methods 201 first episode Chinese Han patients with schizophrenia were given risperidone for up to 56 days.Genotyping of 5-HT2A gene T102C polymorphism were performed by using PCR-RFLP. The Positive and Negative Symptom Scale (PANSS) was used for the evaluation of the severity of psychotic symptoms before and after 8 weeks treatment with risperidone.Results 5-HT2A receptor 102 -T/T genotype was significantly associated with both the PANSS total and negative syndrome subscale scores before treatment, and with the reduction rates in both the PANSS total and negative syndrome subscale scores after eight weeks risperidone treatment.Conclusions The results suggest that 5-HT2A T102C A/A1 genotype subgroup influences individual response to risperidone in first-episode Chinese patients with schizophrenia.

OBJECTIVE: To study the association of the polymorphisms of the serum amyloid A1 (SAA1) gene at rs4638289 and rs7131332 loci with Kawasaki disease (KD) and its complication coronary artery lesion (CAL) in children. METHODS: A total of 105 Han children with KD who were hospitalized and treated from 2013 to 2017 were enrolled as the KD group. A total of 100 Han children who underwent physical examination were enrolled as the control group. According to the presence or absence of CAL, the KD group was further divided into a CAL group with 23 children and a non-CAL (NCAL) group with 82 children. Polymerase chain reaction-restriction fragment length polymorphism was used to investigate the polymorphisms of the SAA1 gene at rs4638289 and rs7131332 loci. RESUKTS: For the locus rs4638289 of the SAA1 gene, there were no significant differences between the KD and control groups in the genotype frequencies of AA, AT, and TT and the allele frequencies of A and T (P>0.05). But there were significant differences between the CAL and NCAL groups in the genotype frequencies of AA, AT, and TT (P=0.016), while there were no significant differences in the allele frequencies of A and T (P>0.05). AT genotype was a protective factor against CAL (OR=0.276, 95%CI: 0.099-0.772, P=0.011). For the locus rs7131332 of the SAA1 gene, there were no significant differences between the KD and control groups in the genotype frequencies of AA, AG, and GG and the allele frequencies of A and G (P>0.05). There were also no significant differences between the CAL and NCAL groups in the genotype frequencies of AA, AG, and GG and the allele frequencies of A and G (P>0.05). CONCLUSIONS Polymorphisms of the SAA1 gene at loci rs4638289 and rs7131332 are not associated with the onset of KD, while the polymorphism at the locus rs4638289 is associated with CAL in KD patients. KD patients with genotype AT may have a reduced risk of CAL.
Case-Control Studies ; Child ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Mucocutaneous Lymph Node Syndrome ; genetics ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Serum Amyloid A Protein ; genetics

OBJECTIVETo investigate the inhibitory effects of saikosaponin-d (SSd) on angiogenesis in chicken embryos and its mechanism of action.

METHODSChorioallantoic membrane (CAM) model was established successfully in 86 chicken embryos. They were divided into 4 groups after fenestration: the three SSd treated groups (A, B and C) treated with high (20 microg/mL, n = 16), middle (10 microg/mL, n = 19) and low (5 microg/mL, n = 25) dose of SSd respectively, and the control group treated with 0.01 mol/L PBS (n = 26). The drug or reagent was administered by grafting 20 microL onto the surface of CAM. After incubation for 3 days, the vessel growth was recorded by digital photography; inflammatory cells were counted under light microscope with HE staining, and the positive rate of angiogenesis reaction was calculated by Leica image analyzer.

RESULTSOn the 6th day of the embryonic age, vessels in the chicken embryo CAM showed a radial growing in spok-wheel pattern around the gelatin sponges with lateral axis running through it. Whereas after 3 days of SSd treatment, the angiogenesis reduced significantly with vague microvessels around the sponge, and vascular truncation and absence revealed. Microscopic examinations showed that the number of microvessels and infiltrated inflammatory cells in the sponge and peripheral CAM mesenchyme in the SSd groups were less than those in the control group, especially on vessels of medium and small size (P < 0.05, P < 0.01, respectively), but was insignificant on great vessels (P > 0.05). Correlation analysis revealed no correlation between the number of the great vessels in CAM and the infiltrated inflammatory degrees (r = 0.117, P > 0.05), but the increase of small vessels in CAM was positively correlated with that of inflammatory cells (r = 0.971, P < 0.01).

CONCLUSIONSSSd could inhibit the physiological angiogenesis of chicken embryoe, especially for the medium and small vessels, while there was no significant effect on great vessels (P > 0.05). Its mechanism of action may be related to its inhibition on leukocyte migration and activation.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Chick Embryo ; Chorioallantoic Membrane ; drug effects ; Neovascularization, Physiologic ; drug effects ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Saponins ; pharmacology