Association of Estrogen Receptor 1 Genetic Polymorphisms with Recurrent Spontaneous Abortion Risk.

BACKGROUND: Estrogen is one of the most important reproductive steroidal hormones and plays a critical role in the maintenance of pregnancy, and its function is mediated by estrogen receptor 1(ESR1). The polymorphisms of ESR1 were involved in recurrent spontaneous abortion (RSA); however, the association between ESR1 polymorphisms and RSA remains controversial. The present meta-analysis was aimed to clarify the association between ESR1 PvuII (-397C/T, rs2234693) and XbaI (-351A/G, rs9340799) polymorphisms and the risk of RSA. METHODS: All the included articles were retrieved from PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, and Wanfang Med Online Database up to January 3, 2018. Data were processed in the Stata 12.0 software. The odds ratios (OR s) and 95% confidence intervals (95% CI s) were calculated using fixed-effects models (FEM)/random-effects models (REM). RESULTS: Seven case-control studies with 836 cases and 1164 controls were included in the study. Generally, the ESR1 polymorphisms were not associated with RSA in any of the genetic analysis models. However, it was found that as rs9340799 polymorphism was related to increased risk of RSA in non-Asian group in the homozygous genetic model (OR = 2.40, 95% CI = 1.05-5.50, P = 0.039). Moreover, in Asian group, rs9340799 polymorphism was found to be related to decreased RSA risk in both the heterozygous model (OR = 0.53, 95% CI = 0.33-0.85, P = 0.009) and the dominant genetic model (OR = 0.55, 95% CI = 0.30-0.98, P = 0.042). CONCLUSIONS: Generally, there was no significant association between the polymorphisms of ESR1 and the risk of RSA. However, subgroup analysis indicated that ESR1 rs9340799 polymorphism was related to increased RSA risk in the non-Asian group while associated with decreased RSA risk in Asian group.

Does Traditional Chinese Medicine pattern affect acupoint specific effect? Analysis of data from a multicenter, randomized, controlled trial for primary dysmenorrhea.

OBJECTIVES: This study assessed the importance of the Traditional Chinese Medicine (TCM) pattern on an acupoint-specific effect. DESIGN: This was a TCM pattern subdivision analysis of the first intervention data from a multicenter, randomized, controlled trial (ISRCTN24863192) (the main trial). SETTINGS: The main trial recruited participants from six hospitals in three provinces in China. SUBJECTS: Five hundred and one (501) participants diagnosed with primary dysmenorrhea (PD) were enrolled in the main trial. INTERVENTIONS: The main trial randomly and equally divided participants into three treatment groups with bilateral electroacupuncture at three sites, respectively: Sanyinjiao (SP6), Xuanzhong (GB39), and an adjacent nonacupoint. Participants were diagnosed with TCM patterns before the treatment. The intervention was carried out when the visual analogue scale (VAS) score of participant's menstrual pain was ≥ 40 mm on the first day of menstruation and lasted for 30 minutes. OUTCOME MEASURES: The immediate improvement of pain was measured with a 100-mm VAS before the intervention, at 5 minutes, 10 minutes, and 30 minutes during the intervention, and at 30 minutes after the completion of this intervention. RESULTS: Three (3) TCM patterns (n=320) were eligible for analysis, including Cold and Dampness Stagnation pattern (n=184), Qi and Blood Stagnation pattern (n=84), and Qi and Blood Deficiency pattern (n=52). In Cold and Dampness Stagnation pattern, the SP6 group had a significant reduction in VAS scores compared with the GB39 group (mean difference -7.6 mm) and the nonacupoint group (mean difference -8.2 mm), respectively. There was no difference between the latter two groups. There were no group differences in VAS scores in the other two patterns. CONCLUSIONS: It suggested that TCM pattern might affect acupoint specific effect on the immediate pain relief obtained for participants with PD.

[Construction of rapid tagging vector of genes in Giardia lamblia].

OBJECTIVE: To construct a recombinant vector for rapid gene tagging in Giardia lamblia. METHODS: To obtain the recombinant vector pGL gdh-Neo with the Neo selection marker, the Neo gene was put under the control of gdh promoter by overlap PCR and inserted into pGEM-5zf. A DNA fragment containing multiple cloning sites (MCS) followed by triple hemagglutinin(3HA) coding sequences was synthesized and cloned into the pGL gdh-Neo to construct a recombinant vector pGL MCS-3HA-gdh-Neo. Giardia H2A gene was selected as a tagging gene to validate the effectivity of the recombinant vector pGL MCS-3HA-gdh-Neo. The histone H2A coding sequence was amplified by PCR, digested with EcoR I and Spe I, and inserted into MCS of pGL MCS-3HA-gdh-Neo. The resulting plasmid was then linearized and transfected into Giardia trophozoites. The H2A recombinant strain selected by G418 was analyzed by PCR,Western blotting and immunofluorescence. RESULTS: A rapid tagging recombinant vector with multiple cloning sites and triple hemagglutinin (3HA) was constructed with a length of 4 260 bp. The H2A recombinant vector was transfected into Giardia trophozoites and integrate into the Giardia genome at the correct locus. The HA-tagged H2A protein was expressed with a molecular weight (Mr) of 16 900. CONCLUSION: A rapid tagging recombinant vector of genes in Giardia lamblia, pGL MCS-3HA-gdh-Neo, has been constructed.

[Cloning, expression and purification of silent information regulator 2 from Giardia lamblia].

OBJECTIVE: To clone and express silent information regulator 2 (Sir2) gene from Giardia lamblia. METHODS: The GlSir2 gene was amplified by PCR from genomic DNA of Giardia lamblia (Chinese strain C2 clone). PCR product was cloned into pMD-19T vector and transformed into E coli JM109. The recombinant plasmid was sequenced and then cloned into the pET28b vector. The pET28b-Gl1Sir2 recombinant plasmid was transformed into E. coli BL21(DE3), followed by expression of the protein induced by IPTG. The recombinant protein was analyzed by SDS-PAGE. Inclusion bodies were dissolved with 8 mol/L urea, and the supernatant was collected and applied to Ni2+ affinity chromatography. The purified recombinant protein was renatured by dialysis and verified by Western blotting using anti-His tag antibody. RESULTS: GlSir2 gene sequence was cloned. The GISir2 open reading frame (1 680 bp) encoded a 559-amino acid protein with Mr 62 800. The recombinant plasmid pET28b-GlSir2 expressed an inclusion body protein of GISir2 after being induced with IPTG. The protein purity reached above 80% after purification. The purified protein was renatured by dialysis. The recombinant GISir2 was recognized by anti-His tag antibody. CONCLUSION: The coding sequence of GLSir2 gene was cloned and expressed in vitro. The recombinant protein was identified by anti-His tag antibody.

[Transcriptional regulation by silence information regulator 2].

Silence information regulator 2 (Sir2) family is a group of conserved deacetylases, widely existed in organisms from archaea to mammals, and characterized by NAD(+)-dependent deacetylase and ADP-ribosyltransferase activities. Sir2 plays an important role in various life progresses, such as chromatin silence, gene regulation, metabolic regulation, life span of cells and so on. Through its deacetylase activity and/or interaction with other proteins, Sir2 can regulate chromatin structure or modify transcription related factors, thus regulating the transcriptional process. This article emphasizes on the progress of Sir2-dependent regulation of gene transcription.