2.Serum progesterone levels in early pregnant women and its relation to luteal maintenance therapy for pregnancy protection
Chinese Journal of General Practitioners 2015;14(4):269-273
Objective To investigate serum levels of β-human chorionic gonadotrophin (β-HCG) and progesterone (P) in early pregnant women,and their relation to luteal maintenance therapy for early pregnancy protection.Methods One hundred and thirty five infertility women treated in Department of Gynecology and Obstetrics of Beijing Chaoyang Hospital from January 2007 to December 2011.Total 150pregnancy cycles,including 84 with intrauterine insemination (IUI) and 66 with natural conception,were divided into two groups:normal intrauterine pregnancy group (group A,n =80) and early pregnancy loss group (group B,n =70).Medical history,ultrasonic findings,serum female hormone,P and β-HCG levels at early pregnancy stage were analyzed.Results The age of group A and group B was (30.0 ± 3.9) years and(30.7 ± 4.9) years,respectively (P > 0.05).The follicle-stimulating hormone/luteinizing hormone in group A and group B was 1.57 ±0.96 and 1.56 ± 1.08 ; the estradiol levels in two groups were (152 ±66) pmol/L and (147 ± 69) pmol/L,respectively (both P > 0.05).There were no differences in dominant follicles and endometrial thickness between groups A and B (P > 0.05).Ovulation promotion and luteal support treatments were adopted in both groups:50% (40/80) of cycles in group A received ovulation promotion,73% (58/80) of cycles received luteal support,while 44% (31/70) and 76% (53/70) received in groups B,respectively.The levels of serum progesterone in group A during 14-21 d,22-27 d and ≥ 28 d after ovulation were higher than those in group B at each time points (P < 0.0l).There were no significant differences in serum progesterone levels between women with luteal support treatment and those without luteal support treatment in both groups (P > 0.05).Conclusion Dynamic monitoring of serum progesterone level in early pregnancy may be used as an auxiliary index for prediction of pregnancy outcome,but it may not be an indicator for luteal maintenance therapy.
3.Evaluation of clinical characteristics of genitourinary syndrome in postmenopausal women
Jiao CHENG ; Biao WANG ; Zhenyu ZHANG ; Xiaodong ZHANG
Chinese Journal of General Practitioners 2016;15(7):553-556
One hundred and ten postmenopausal women,complaining vaginal discomfort,recurrent urinary tract infections or painful intercourse and visiting the hospital between March 2014 and February 2015 were enrolled in the study.Among them,58 patients received vaginal administration of estriol estriol ointment (group A) and 52 patients received conventional treatment (group B).There were no significant differences in mean age of menopause and mean menopause duration between two groups.The clinical symptoms,vaginal health scores,pelvic organ prolapse were observed after treatment.After 6-month treatment,the female vaginal health score was significantly improved in women group A,the rate of urinary orifice was reduced (78% vs.7%),and the rate of pelvic organ prolapse was decreased (36% vs.21%).The geritourinary syndrome of menopause is a new term for vulvovaginal atrophy with clinical characteristics of multi-system changes in postmenopausal women,including atrophic vaginitis,female urogenital diseases,and pelvic organ prolapse.The new diagnostic criteria are needed for evaluation and management of geritourinary syndrome.
4.Comparison between Astragalus membranaceus var. mongholicus and Hedysarum polybotrys based on ITS sequences and metabolomics.
Meili JIAO ; Zhenyu LI ; Fusheng ZHANG ; Xuemei QIN
Acta Pharmaceutica Sinica 2015;50(12):1625-31
Astragalus membranaceus var. mongholicus and Hedysarum polybotrys belong to different genera, but have similar drug efficacy in traditional Chinese medicine theory, and H. polybotrys was used as the legal A. membranaceus var. mongholicus previously. In this study, similarities and differences between them were analyzed via their ITS/ITS2 fragments information. The ITS (internal transcribed spacer) regions were amplified using polymerase chain reaction and then sequenced in two-way. The alignment lengths of ITS regions were 616 bp, in which 508 loci were consistent, and 103 loci were different, accounting for 82.47% and 16.72% of the total ITS nucleotides in length, respectively. As genotype determines phenotype, 1HNMR-based metabolomic approach was further used to reveal the chemical similarities and differences between them. Thirty-four metabolites were identified in the 1H NMR spectra, and twenty-seven metabolites were the common components. Amino acids, carbohydrates and other primary metabolites were similar, while a large difference existed in the flavonoids and astragalosides. This study suggests that A. membranaceus var. mongholicus and H. polybotrys show similarities and differences from molecular and chemical perspectives, which has laid a foundation for elucidating the effective material basis of drug with similar efficacy and resources utilization.
6.Protective effect of osthole on SH-SY5Y cells transfected with APP595/596 gene
Yanan JIAO ; Yingjia YAO ; Liang KONG ; Shaoheng LI ; Zhenyu TAO ; Yuhui YAN ; Jingxian YANG
Chinese Journal of Pathophysiology 2015;(11):2053-2058
AIM:To explore the protective effect of osthole on the SH-SY5Y cells transfected with APP595/596 gene, and to investigate the molecular mechanism.METHODS:The SH-SY5Y cells were transfected with APP595/596 gene in vitro for establishing a cell model to study the pathogenic role of amyloid β-protein ( Aβ) .The cell viability was detected by CCK-8 assay.The release of lactate dehydrogenase ( LDH) was determined by the colour reaction of dia-phorase-INT.The cell apoptotic rate was analyzed by flow cytometry.The expression of β-site APP cleaving enzyme 1 ( BACE1) at mRNA and protein levels was detected by RT-PCR and Western blot.The expression of Aβwas measured by the technique of immunofluorescence cytochemistry and Western blot.RESULTS: Treatment with osthole inhibited the LDH release, and increased the viability of the cells.The percentage of apoptotic cells was also significantly decreased. Osthole also inhibited the expression of BACE1 at mRNA and protein levels and the protein expression of Aβ.CONCLU-SION:Osthole has protective effect on SH-SY5Y cells transfected with APP595/596 gene.The mechanism may be associ-ation with inhibiting the mRNA and protein expression of BACE1.
7.Osthole promotes the proliferation of neural stem cells in vitro
Yingjia YAO ; Yu HU ; Shaoheng LI ; Yanan JIAO ; Liang KONG ; Zhenyu TAO ; Jingxian YANG
Chinese Journal of Tissue Engineering Research 2014;(32):5184-5189
BACKGROUND:Neural stem cells have self-renewal and multidirectional differentiation potential, but under normal circumstances, the number of neural stem cells is less, and most cells are in the resting state. Thus, to promote the proliferation of neural stem cells is the key to the treatment of neurodegenerative diseases. OBJECTIVE:To investigate the effects of osthole on the proliferation of neural stem cells cultured in vitro, and to analyze its mechanism underlying promoting the proliferation. METHODS:Neural stem cells were cultured in vitro, and passage 3 cells were cultured with different concentrations of osthole(10, 50 and 100μmol/L). After 24 hours, cellvitality was determined by cellcounting kit-8. After 3, 5, 7 days of further culture, the radius of neurospheres was measured, and Ki67-positive cells were counted by immunofluorescence staining. Meanwhile, after 3 days of further culture, the gene expression of Notch 1, Hes 1 and Mash 1 in neural stem cells was detected by RT-PCR. RESULTS AND CONCLUSION:Compared with the control group, 50, 100μmol/L osthole could obviously promote the proliferation ability of neural stem cells. 100μmol/L osthole had the most significant effect and increased the expression of Notch 1 gene, Hes 1 gene, but it had no effect on Mash 1 gene. These results suggest that osthole can promote proliferation of neural stem cells cultured in vitro and its mechanism may be associated with activation of Notch 1 gene and Hes 1 gene in Notch signaling pathway.
8.Anti-apoptosis and anti-inflammatory effect of osthole in mice following stab wound injury
Liang KONG ; Yingjia YAO ; Yanan JIAO ; Shaoheng LI ; Zhenyu TAO ; Yuhui YAN ; Jingxian YANG
Chinese Pharmacological Bulletin 2015;(7):999-1003,1004
Aim To investigate the effects of osthol on cell apoptosis and inflammatory cell infiltration after brain stab wound injury in mice. Methods The mice underwent the stab wound injury by a needle, then were randomly divided into sham operation group, model group, osthol 10, 20, 30 mg · kg-1 treatment group. The main examinations included mice brain wa-ter content; the apoptotic cytokines Bax, Bcl-2, Caspase-3 mRNA expression were assessed by PT-PCR; immunohistochemistry staining was used to de-tect neutrophils (MPO) and microglia (Iba-1) infiltra-tion and Caspase-3 positive cell expression around in-jured lesions. Results Treatment with osthole 20, 30 mg·kg-1 group significantly reduced the water content in injured brain, improved the ratio of Bax/Bcl-2, and reduced the expression of apoptosis cytokine Caspase-3 mRNA. Osthole 30 mg·kg-1 treatment group obvious-ly reduced the infiltration of neutrophils and microglial cells and significantly reduced the number of apoptotic cells around the injured cerebral cortex. Conclusion Osthole has therapeutic effect on stab wound injury in mice, and the possible mechanism may be by reducing the infiltration of inflammatory cells and reducing apop-totic cells.
9.Neurotrophin 3 gene overexpression promotes cholinergic differentiation in neural stem cells
Yuhui YAN ; Shaoheng LI ; Liang KONG ; Yingjia YAO ; Yanan JIAO ; Zhenyu TAO ; Jie SONG ; Jingxian YANG
Chinese Pharmacological Bulletin 2016;32(5):631-637
Aim To investigate the effects of neurotro-phin-3 ( NT-3 ) gene overexpression on the differentia-tion into cholinergic neuron of neural stem cells ( NSCs) in vitro and its underlying mechanism. Meth-ods Brain-derived NSCs from newborn mice were iso-lated and cultured in vitro and determined by immuno-fluorescence. The NSCs were divided into three groups: NSCs, GFP-NSCs and NT-3-NSCs groups. The expression of NT-3 was detected by immunofluo-rescence and ELISA. Then, the ability of NSCs on dif-ferentiation into cholinergic neuron was detected by im-munofluorescence and RT-PCR, and the Acetylcholine Assay Kit was used for acetylcholine ( ACh) , and the expression of Hes1 , Mash1 and Ngn1 mRNA was de-termined by RT-PCR. Results The neurosphere dis-played Nestin and Sox 2-postive by immunofluores-cence, suggesting that the cultured cells were NSCs. The proportion of ChAT immunopositive cells was sig-nificantly higher in the NT-3-NSCs group than that in the other two groups ( P <0. 01 ) . Ach secretion in NT-3-NSCs was significantly elevated compared with the other two groups ( P <0. 01 ) . NSCs transfected with NT-3 increased the levels of Mash1 and Ngn1 mR-NA, and decreased the level of Hes1 mRNA ( P <0. 05 ) . Conclusion NT-3 can significantly promote the in vitro differentiation of NSCs into cholinergic neu-rons via probablly inhibiting Notch signaling pathway.
10.Osthole promotes differentiation into neurons and reduces neuronal apoptosis via Wnt/β-catenin signaling pathway in APP transduced neural stem cells
Yingjia YAO ; Liang KONG ; Yanan JIAO ; Shaoheng LI ; Zhenyu TAO ; Yuhui YAN ; Jingxian YANG
Chinese Pharmacological Bulletin 2015;(11):1516-1523
Aim To investigate the effects of osthole ( Ost) on the ability of proliferation and differentiation in APP transduced neural stem cells( NSCs) , and neu-ronal apoptosis, in order to find related mechanism. Methods A model of Alzheimer′s disease( AD) cells was successfully established by transducing APP gene into NSCs in vitro. The ability of proliferation and dif-ferentiation was tested by staining. The viability of NSCs was determined by using CCK-8 assay. The cell apoptosis was tested by Hoechst 33258 staining. The expression of GSK-3β and β-catenin mRNA was deter-mined by RT-PCR. The expression of GSK-3β and β-catenin protein was determined by Western blot. Re-sults The ability of proliferation had increased by 10 . 24% with Ost treatment, compared with APP group. The ability of differentiation had increased by 6 . 74%with Ost treatment, compared with APP group. The vi-ability of NSCs had increased and cell apoptotic rate had decreased significantly. From the results of RT-PCR and Western blot, we could find the expression of GSK-3βmRNA and protein had decreased, and the ex-pression of β-catenin mRNA and protein had increased significantly, compared with APP group. Conclusion Ost could enhance the ability of proliferation and dif-ferentiation into more neurons of NSCs transducing APP gene, and reduce neuronal apoptosis. It might be relat-ed with activiting Wnt/β-catenin signaling pathway.