To analyze and compare the protective effects of active components in different ethyl acetate extracts (EAEEPs) from Eclipta prostrate, in order to study the comparison of materials bases protecting normal human bronchial epithelial (NHBE) cells. The MTT assay was taken to compare the protective effect of different EAEEPs on cigarette smoke extracts (CSE) -induced NHBE cells. The ultra-performance liquid chromatography (UPLC) was applied to analyze the content of phenolic acid, coumaric grass ether and flavonoid in EAEEPs. According to the results, all of the eight EAEEPs (0-200 mg x L(-1)) showed certain protective effect on NHBE cells, with statistical difference. Specifically, the total mass of EAEEP VII (89.15 mg x L(-1)) and EAEEP VIII (57.44 mg x L(-1)), which showed the strongest activity, was not the highest, while EAEEP III (132.25 mg x L(-1)) displayed the highest total mass. In the combination with the "component structure" theory, the analysis showed a significant difference in the mass structure among phenolic acid, coumaric grass ether and flavonoid in EAEEP VIII and EAEEP VIII, which were 1.0: 1. 0: 0.5 and 1.0: 1.9: 0.8, respectively. The results suggested a specific optimal "component structure" relationship may exist in EAEEP, which could provide reference for the material base study and quality control.
Bronchi ; cytology ; drug effects ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Eclipta ; chemistry ; Epithelial Cells ; cytology ; drug effects ; Humans ; Protective Agents ; chemistry ; isolation & purification ; pharmacology ; Tobacco Smoke Pollution ; adverse effects

Objective To investigate the inhibitory effects of RNA interference(RNAi)on the expression of matrix metalloproteinase-9(MMP-9)gene and invasiveness and adhesion of ovarian cancer cells.Methods Four groups of different specific target sequence in coding region of MMP-9 and one non- specific sequence were chosen,which were Sitel,Site2,Site3,Site4 and Site5.Small interference RNA (siRNA)expression cassettes(SEC)were constructed by PCR and transfected into ovarian cancer HO- 8910PM cells.RT-PCR and western blot were used to detect mRNA and protein expression of MMP-9 gene; the abilities of invasion and adhesion were detected by Matrigel invasion assay and cell adhesion assay. Results The expression of MMP-9 was inhibited and the inhibitory effects of different sequence were varied.The mRNA expression was 0.64?0.06,0.47?0.07,0.55?0.10 in Sitel,Site2,Site3 group, and protein expression was 0.30?0.09,0.27?0.08,0.37?0.12,respectively.Site2 group had the most efficient inhibitory effect,followed by Sitel and Site3 groups.Cell growth curve revealed that cell growth was significantly inhibited in Site2 group.Invasiveness and adhesion were significantly reduced,the inhibitory rate on invasion in Site1,Site2,Site3 groups were 50.0%,50.0% and 37.5%,respectively;the inhibitory rate on adhesion in Site1,Site2,Site3,Site4 groups were 43.8%,48.8%,33.9%,24.2% at 60 min and 41.6%,40.2%,35.1%,16.0% at 90 min,respectively.Conclusions RNAi exists in ovarian HO-8910PM cells.MMP-9 siRNA can specifically down-regulate MMP-9 expression and lead to the inhibition of invasiveness and adhesion in ovarian cancer cells.

To introduce the application of Stata software to heterogeneity test in meta-analysis.A data set was set up according to the example in the study,and the corresponding commands of the methods in Stata 9 software were applied to test the example.The methods used were Q-test and Ⅰ2statistic attached to the fixed effect model forest plot,H statistic and Galbraith plot.The existence of the heterogeneity among studies could he detected hy Q-test and H statistic and the degree of the heterogeneity could be detected by,Ⅰ2 statistic.The outliers which were the sources of the heterogeneity could be spotted from the Galbraith plot.Heterogeneity test in Meta-analysis can be completed by the four methods in Stata software simply and quickly.H and Ⅰ2 statistics are more robust,and the outliers of the heterogeneity can be clearly seen in the Galbraith plot among the four methods.

The difference between three representative components of total salvianolic acids in pharmacodynamic activity were compared by three different pharmacological experiments: HUVECs oxidative damage experiment, 4 items of blood coagulation in vitro experiment in rabbits and experimental myocardial ischemia in rats. And the effects of contribution rate of each component were calculated by multi index comprehensive evaluation method based on CRITIC weights. The contribution rates of salvianolic acid B, rosmarinic acid and Danshensu were 28.85%, 30.11%, 41.04%. Apparent oil/water partition coefficient of each representative components of total salvianolic acids in n-octyl alcohol-buffer was tested and the total salvianolic acid components were characterized based on a combination of the approach of self-defined weighting coefficient with effects of contribution rate. Apparent oil/water partition coefficient of total salvianolic acids was 0.32, 1.06, 0.89, 0.98, 0.90, 0.13, 0.02, 0.20, 0.56 when in octanol-water/pH 1.2 dilute hydrochloric acid solution/ pH 2.0, 2.5, 5.0, 5.8, 6.8, 7.4, 7.8 phosphate buffer solution. It provides a certain reference for the characterization of components.
Animals ; Benzofurans ; chemistry ; pharmacology ; Cinnamates ; chemistry ; pharmacology ; Depsides ; chemistry ; pharmacology ; Lactates ; chemistry ; pharmacology ; Male ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Solubility

OBJECTIVETo study the effect of Tangshenkang Granule (TG) containing serum on renal mesangial cells' (RMCs) proliferation and TGF-β1/Smad2/3 pathway in the high glucose condition.

METHODSTwelve SD rats were randomly divided into four groups, i.e., the low dose TG group, the middle dose TG group, the high dose TG group, and the blank control group, 3 in each group. After 7-day gastrogavage via portal vein blood, rats were sacrificed and their serum samples were collected. RMCs were cultured in common rat serum and TG containing serum respectively. The proliferation of mesangial cells was determined by methly thiazolyl tetrazolium (MTT) assay to determine the optimal TG containing serum concentration. Expression levels of TGF-β1 mRNA and protein were determined by real time quantitative PCR and ELISA. Smad2/3 protein expression and phosphorylation were determined by Western blot and immunofluorescence.

RESULTSTG containing serum at different doses could inhibit high glucose induced RMC cells' proliferation, TGF-β1 over-expression and Smad2/3 phosphorylation.

CONCLUSIONTG containing serum could inhibit high glucose induced RMC cells' proliferation, and its mechanism might be possibly associated with inhibiting TGF-β1/Smad2/3 signaling pathway.

Animals ; Cell Proliferation ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; Mesangial Cells ; Phosphorylation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Serum ; Signal Transduction ; Smad2 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo observe the effect of Yangjing Zhongyu Decoction (YZD) on mRNA and protein expression of PCNA, StAR, and FSHR in ovarian granulose cells (GCs) cultured by excess androgen.

METHODSOvarian GCs from porcine follicles were isolated and cultured in vitro. Follicular stimulating hormone (FSH) or YZD was added in the GCs treated by excess testosterone propionate. Totally 48 h later mRNA and protein expression of PCNA, StAR, and FSHR were detected by RT-PCR and Western blot.

RESULTSExcess androgen inhibited mRNA and protein expression of PCNA, StAR, and FSHR of GCs. FSH and YZD could antagonize inhibition of excess androgens, and promote mRNA and protein expression of PCNA, StAR, and FSHR in GCs.

CONCLUSIONYZD could antagonize the inhibition of excess androgen on mRNA and protein expression of PCNA, StAR and FSHR in GCs. Thus, we inferred that YZD could improve the follicle dysplasia by promoting mRNA and protein expression of PCNA, StAR and FSHR in GCs.

Androgens ; pharmacology ; Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Follicle Stimulating Hormone ; pharmacology ; Granulosa Cells ; cytology ; drug effects ; metabolism ; Membrane Transport Proteins ; genetics ; metabolism ; Ovarian Follicle ; cytology ; drug effects ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; RNA, Messenger ; genetics ; Receptors, FSH ; genetics ; metabolism ; Swine

OBJECTIVETo investigate the interaction between myeloid-derived suppressor cells (MDSCs) and nature killer cells during acute fulminate hepatitis.

METHODSAcute fulminate hepatitis were induced by i.p. co-injection of LPS and D-GalN in mice, and the ratio of MDSCs,NK cells and the activation of NK cells in different tissues were analyzed by FACS at 0 h,1.5 h,3 h and 6 h.

RESULTSThe percentage of MDSCs and NKG2D+NK cells in different tissues increased as acute fulminate hepatitis progressed, with the increased NK cells in liver tissue. The mean fluorescence intensity of NKG2D on NK cells in different tissues were also enhanced.

CONCLUSIONMDSCs induce the proliferation and activation of NK cells in mice with acute fulminate hepatitis.

Acute Disease ; Animals ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Hepatitis ; etiology ; immunology ; Killer Cells, Natural ; immunology ; Lipopolysaccharides ; Liver Failure, Acute ; chemically induced ; immunology ; Male ; Mice ; Mice, Inbred C57BL ; Myeloid Cells ; immunology ; T-Lymphocytes, Regulatory ; immunology

Humans ; Infant, Newborn ; Male ; Nephrotic Syndrome ; congenital ; Syphilis, Congenital ; complications ; diagnosis ; drug therapy

This study aimed to explore the outcomes of progestin-primed ovarian stimulation protocol (PPOS) in aged infertile women who failed to get pregnant in the first IVF/ICSI-ET cycles with GnRH-a long protocol.A self-controlled study was conducted to retrospectively investigate the clinical outcomes of 104 aged infertile patients who didn't get pregnant in the first IVF/ICSI-ET treatment by stimulating with GnRH-a long protocol (non-PPOS group),and underwent PPOS protocol (PPOS group) in the second cycle between January 2016 and December 2016 in the Center for Reproductive Medicine,Renmin Hospital of Wuhan University.The primary outcomes included clinical pregnancy rate of frozen-thawed embryos transfer (FET) in PPOS group,and good-quality embryo rate in both groups.The secondary outcomes were fertilization rate,egg utilization rate and cycle cancellation rate.The results showed that there were no significant differences in basal follicle stimulating hormone (bFSH),antral follicle count (AFC),duration and total dosage of gonadotropin (Gn),number of oocytes retrieved,intracytoplasmic sperm injection (ICSI) rate,fertilization rate,and cycle cancellation rate between the two groups (P＞0.05).However,the oocyte utilization rate and good-quality embryo rate in PPOS group were significantly higher than those in non-PPOS group (P＜0.05).By the end of April 2017,62 FET cycles were conducted in PPOS group.The clinical pregnancy rate and embryo implantation rate were 22.58％ and 12.70％,respectively.In conclusion,PPOS protocol may provide better clinical outcomes by improving the oocyte utilization rate and good-quality embryo rate for aged infertile patients who failed to get pregnant in the first IVF/ICSI-ET cycles.

Our previous studies showed that the proto-oncogene c-erbB₂ played an important role in primordial follicles growth. The present study was conducted to investigate the role of MAPK and PKC signaling pathways in the primordial follicle onset in neonatal rats, and the relationship between c-erbB₂ and MAPK/PKC signaling pathways. Ovaries collected from 2-day-old Sprague-Dawley rats were cultured in the Waymouth culture system in vitro. Ovaries were transfected with c-erbB₂ siRNA, or treated with PD98059 (50 mumol/L) or Calphostin (0.5 mumol/L) in the culture medium. RT-PCR was performed to measure the expression of c-erbB₂ mRNA, and Western blot analysis was performed to measure the expression of ErbB₂, MAPK and PKC protein after the neonatal rat ovaries were cultured for 8 d. The quantities of every-stage follicles of ovaries cultured for 8 d were obtained in histological section stained with hematoxylin eosin. The results showed that c-erbB₂ siRNA reduced the levels of c-erbB₂ mRNA (P<0.01) and the levels of ErbB₂, MAPK and PKC protein (P<0.01) significantly. But the levels of c-erbB₂ mRNA and ErbB₂ protein exhibited no change (P>0.05) in the ovaries cultured with PD98059 or Calphostin. After the ovaries were transfected with c-erbB₂ siRNA or cultured with PD98059 or Calphostin for 8 d, the quantities of primary follicles and second follicles were lower than those in the control group (P<0.05 or P<0.01), but the quantity of the primordial follicles was higher than that in the control group (P<0.01). These results suggest that proto-oncogene c-erbB₂ promotes the initiation of primordial follicle growth through the MAPK and PKC signal transduction, and c-erbB₂ is possibly the upstream of PKC and MAPK signaling pathway in the regulation of primordial follicle onset.
Animals ; Animals, Newborn ; Enzyme Inhibitors ; pharmacology ; Female ; Flavonoids ; pharmacology ; MAP Kinase Signaling System ; Naphthalenes ; pharmacology ; Ovarian Follicle ; metabolism ; Protein Kinase C ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; Rats ; Rats, Sprague-Dawley ; Receptor, ErbB-2 ; metabolism ; Signal Transduction