Objective:To observe the ameliorative effects of exogenous miR-146a-5p on lipopolysaccharide (LPS)-induced embryonic resorption and fetal mouse dysplasiamice, and to preliminarily investigate its mechanism of action.Methods:1) After 36 healthy adult female mice were mated with male mice, uterine tissues were collected from females on day (D) 0 (D0/not pregnant), D0.5 (the day of embryo observed), D4.5, D7.5, D9.5 and D13.5 of gestation, and the expression levels of miR-146a-5p and its target gene TRAF6 protein in uterine tissues of mice at different gestation periods were detected by real-time fluorescent quantitative PCR (qPCR) and Western blotting. 2) The mice on D7.5 of pregnancy were treated with intraperitoneal injection of saline (control, COL group), intraperitoneal injection of 250 μg/kg LPS (named LPS250 group), LPS combined with tail vein injection of 10 nmol miR-146a-5p unrelated sequence (negative control, NC, named LPS250+NC group), or LPS combined with tail vein injection of 10 nmol miR-146a-5p agonist (miR-146a-5p agomir, named LPS250+miR-146a-5p agomir group). The total number of embryos and the number of absorbed embryos in the uterus of pregnant mice were measured and statistically analyzed on D8.5, and the expression levels of TNFα mRNA and TRAF6 protein in uterine tissues were detected by qPCR and Western blotting. 3) Then we reduced the dosage of LPS to 50 μg/kg and treated the same groups, named LPS50+NC group, LPS50+miR-146a-5p agomir group, respectively. The total number of fetal mice/embryos, the number of absorbed embryos, the number of surviving fetal mice, the weight of surviving fetal mice and the weight of the placenta were measured and statistically analyzed on D16.5. 4) Primary mouse bone marrow-derived macrophages (BMDM) were isolated and cultured. Mouse BMDM was inducted to M1 polarization by LPS stimulation, and then was transient transfected miR-146a-5p mimics or their NC fragments. The expression levels of TNFα mRNA and pSTAT1 protein were detected by qPCR and Western blotting. Results:The expression level of miR-146a-5p was significantly higher in the implantation sites of D7.5, D9.5 and D13.5 pregnant mice than in the non-implantation sites ( P=0.013, P=0.012, P=0.003), and the protein expression level of TRAF6 was significantly lower in the implantation site of D13.5 pregnant mice than in the non-implantation site ( P=0.012). After intraperitoneal injection of 250 μg/kg of LPS into D7.5 pregnant mice, the embryo absorption rate of the LPS group on D8.5 was 43.13%±3.31%, which was significantly higher than that of COL group (0%, P=0.002), while the embryo absorption rate of the LPS250+miR-146a-5p agomir group (13.50%±0.87%) was significantly lower than that of the LPS250+NC group (59.33%±4.04%, P=0.001). After intraperitoneal injection of 50 μg/kg of LPS combined with tail vein injection of NC or miR-146a-5p agomir to D7.5 pregnant mice, the fetal mouse weight [(0.29±0.09) g] and placental weight [(0.06±0.02) g] of surviving fetal mice in the LPS50+NC group on D16.5 and the LPS50+miR-146a-5p agomir group were statistically significant [(0.46±0.06) g, P<0.001; (0.07±0.02) g, P=0.021], and the differences in the number of absorbed embryos and embryo uptake rate between the two groups were not statistically significant (all P>0.05). The expression levels of both pSTAT1 protein and TNFα mRNA were significantly downregulated in BMDM transfected with miR-146a-5p mimics compared with those transfected with NC ( P=0.012, P=0.039). Conclusion:miR-146a-5p expression levels were significantly increased at the maternal-fetal interface during the late stage of mouse embryo implantation and placental development. Exogenous miR-146a-5p could effectively improve LPS-induced mouse embryo resorption and fetal mouse dysplasia. miR-146a-5p could inhibit the M1 polarization activity of mouse macrophages, suggesting that miR-146a-5p may inhibit the M1 polarization activity of mouse macrophages by suppressing M1 polarization of mouse maternal-fetal interface macrophages to safeguard the normal establishment and maintenance of pregnancy.

Objective:To observe the ameliorative effects of exogenous miR-146a-5p on lipopolysaccharide (LPS)-induced embryonic resorption and fetal mouse dysplasiamice, and to preliminarily investigate its mechanism of action.Methods:1) After 36 healthy adult female mice were mated with male mice, uterine tissues were collected from females on day (D) 0 (D0/not pregnant), D0.5 (the day of embryo observed), D4.5, D7.5, D9.5 and D13.5 of gestation, and the expression levels of miR-146a-5p and its target gene TRAF6 protein in uterine tissues of mice at different gestation periods were detected by real-time fluorescent quantitative PCR (qPCR) and Western blotting. 2) The mice on D7.5 of pregnancy were treated with intraperitoneal injection of saline (control, COL group), intraperitoneal injection of 250 μg/kg LPS (named LPS250 group), LPS combined with tail vein injection of 10 nmol miR-146a-5p unrelated sequence (negative control, NC, named LPS250+NC group), or LPS combined with tail vein injection of 10 nmol miR-146a-5p agonist (miR-146a-5p agomir, named LPS250+miR-146a-5p agomir group). The total number of embryos and the number of absorbed embryos in the uterus of pregnant mice were measured and statistically analyzed on D8.5, and the expression levels of TNFα mRNA and TRAF6 protein in uterine tissues were detected by qPCR and Western blotting. 3) Then we reduced the dosage of LPS to 50 μg/kg and treated the same groups, named LPS50+NC group, LPS50+miR-146a-5p agomir group, respectively. The total number of fetal mice/embryos, the number of absorbed embryos, the number of surviving fetal mice, the weight of surviving fetal mice and the weight of the placenta were measured and statistically analyzed on D16.5. 4) Primary mouse bone marrow-derived macrophages (BMDM) were isolated and cultured. Mouse BMDM was inducted to M1 polarization by LPS stimulation, and then was transient transfected miR-146a-5p mimics or their NC fragments. The expression levels of TNFα mRNA and pSTAT1 protein were detected by qPCR and Western blotting. Results:The expression level of miR-146a-5p was significantly higher in the implantation sites of D7.5, D9.5 and D13.5 pregnant mice than in the non-implantation sites ( P=0.013, P=0.012, P=0.003), and the protein expression level of TRAF6 was significantly lower in the implantation site of D13.5 pregnant mice than in the non-implantation site ( P=0.012). After intraperitoneal injection of 250 μg/kg of LPS into D7.5 pregnant mice, the embryo absorption rate of the LPS group on D8.5 was 43.13%±3.31%, which was significantly higher than that of COL group (0%, P=0.002), while the embryo absorption rate of the LPS250+miR-146a-5p agomir group (13.50%±0.87%) was significantly lower than that of the LPS250+NC group (59.33%±4.04%, P=0.001). After intraperitoneal injection of 50 μg/kg of LPS combined with tail vein injection of NC or miR-146a-5p agomir to D7.5 pregnant mice, the fetal mouse weight [(0.29±0.09) g] and placental weight [(0.06±0.02) g] of surviving fetal mice in the LPS50+NC group on D16.5 and the LPS50+miR-146a-5p agomir group were statistically significant [(0.46±0.06) g, P<0.001; (0.07±0.02) g, P=0.021], and the differences in the number of absorbed embryos and embryo uptake rate between the two groups were not statistically significant (all P>0.05). The expression levels of both pSTAT1 protein and TNFα mRNA were significantly downregulated in BMDM transfected with miR-146a-5p mimics compared with those transfected with NC ( P=0.012, P=0.039). Conclusion:miR-146a-5p expression levels were significantly increased at the maternal-fetal interface during the late stage of mouse embryo implantation and placental development. Exogenous miR-146a-5p could effectively improve LPS-induced mouse embryo resorption and fetal mouse dysplasia. miR-146a-5p could inhibit the M1 polarization activity of mouse macrophages, suggesting that miR-146a-5p may inhibit the M1 polarization activity of mouse macrophages by suppressing M1 polarization of mouse maternal-fetal interface macrophages to safeguard the normal establishment and maintenance of pregnancy.

Objectives:To screen out genes potentially involved in the dysregulation of immune microhomeostasis at the maternal-fetal interface of recurrent miscarriage (RM) patients, and to identify novel biomarkers of RM by bioinformatic analysis.Methods:The dataset GSE165004 of endometrial tissues from RM patients ( n=24) and normal women as the control ( n=24) was downloaded from the GEO database, and differentially expressed genes (DEGs) and immune-related modules were analyzed by using the R language's Limma package, along with CIBERSORT immune infiltration and Weighted Gene Co-expression Network Analysis (WGCNA). The functional associations of these core genes were evaluated through Gene Set Enrichment Analysis (GSEA) and Gene Set Variation Analysis (GSVA). Finally, we used the decidual tissue dataset GSE161969 to further validate the diagnostic value of these key genes. Results:Differential analysis identified 580 DEGs, and 3 271 immune-related modular genes were selected by WGCNA analysis. FGF2, ANO1, and LAPTM5 were subsequently identified as key genes through machine learning techniques. GSVA analysis further revealed critical roles of FGF2, ANO1 and LAPTM5 in immune infiltration and macrophage pathways. Conclusion:FGF2, ANO1 and LAPTM5 might participate in the immuno-related pathogenesis of RM, and present potential biomarkers for the early diagnosis and treatment of RM.

Objectives:To screen out genes potentially involved in the dysregulation of immune microhomeostasis at the maternal-fetal interface of recurrent miscarriage (RM) patients, and to identify novel biomarkers of RM by bioinformatic analysis.Methods:The dataset GSE165004 of endometrial tissues from RM patients ( n=24) and normal women as the control ( n=24) was downloaded from the GEO database, and differentially expressed genes (DEGs) and immune-related modules were analyzed by using the R language's Limma package, along with CIBERSORT immune infiltration and Weighted Gene Co-expression Network Analysis (WGCNA). The functional associations of these core genes were evaluated through Gene Set Enrichment Analysis (GSEA) and Gene Set Variation Analysis (GSVA). Finally, we used the decidual tissue dataset GSE161969 to further validate the diagnostic value of these key genes. Results:Differential analysis identified 580 DEGs, and 3 271 immune-related modular genes were selected by WGCNA analysis. FGF2, ANO1, and LAPTM5 were subsequently identified as key genes through machine learning techniques. GSVA analysis further revealed critical roles of FGF2, ANO1 and LAPTM5 in immune infiltration and macrophage pathways. Conclusion:FGF2, ANO1 and LAPTM5 might participate in the immuno-related pathogenesis of RM, and present potential biomarkers for the early diagnosis and treatment of RM.

Objective To analyse the high-risk human papilloma virus (HPV) infection in women, and to analyze the protective effect of bivalent HPV vaccine on HPV infection. Methods A case-control study method was used to retrospectively investigate the HPV infection status of 9246 women who received high-risk HPV infection examination in the outpatient department of Shiyan people's Hospital of Hubei from January 2018 to December 2018. The second-generation hybrid capture method and colposcopy examination were used to diagnose. Using a 1:1 matching method, the uninfected individuals who were examined during the same period were taken as the control group, and the confirmed infected group was taken as the case group, and the differences in the vaccination rates of the bivalent HPV vaccine between the two groups were compared. At the same time, the case group was divided into two groups according to the vaccinated and unvaccinated groups and followed up. The events ranged from 24 to 36 months. The incidence of persistent HPV infection, cervical intraepithelial neoplasia (CIN) and cervical cancer were counted to understand Protective effect of bivalent HPV vaccination against HPV infection in a high-risk female population. Results A total of 1 632 cases (17.65%) of 9 246 women were screened positive for high-risk HPV infection. Chi-square results showed that the HPV positive infection rate of rural women (32.84%) was lower than that of urban women (67.16%). , Marital status also has a certain influence on HPV infection. Among the 1632 cases of HPV positive infection, 629 cases (38.54%) were vaccinated with bivalent HPV vaccine, and 1003 cases (61.46%) were not vaccinated with bivalent HPV vaccine. During the follow-up period of 24-36 months, the vaccination group finally obtained follow-up data of 584 cases due to unwillingness to cooperate (18 cases), unable to conduct research due to organic changes (24 cases), and mental disorders (3 cases), with a loss to follow-up rate of 7.15 cases. %; In the unvaccinated group, 949 cases of follow-up data were finally obtained due to change of residence (32 cases), low degree of cooperation (20 cases) and psychological factors (2 cases), and the loss to follow-up rate was 5.38%. The results after follow-up showed that the persistent HPV infection rate in the bivalent HPV vaccination group, the positive rate of high-risk HPV infection at the last follow-up, the cumulative incidence of CIN1 during the follow-up period, the cumulative incidence of CIN2+ during the follow-up period, the incidence of CIN1 at the last follow-up, and the incidence of CIN2+ at the last follow-up. and cervical cancer incidence rates were 3.07%, 0.82%, 1.84%, 1.02%, 0.82%, 0.20%, and 0.00%, respectively, and the bivalent HPV unvaccinated groups were 12.91%, 15.52%, 7.14%, 4.40%, and 3.02%, respectively. , 1.37% and 0.27%. Persistent HPV infection rate, positive rate of high-risk HPV infection at last follow-up, cumulative incidence of CIN1 during follow-up, cumulative incidence of CIN2+ during follow-up, incidence of CIN1 at last follow-up, and incidence of CIN2+ at last follow-up were significantly lower in bivalent HPV vaccination group in the control group (P<0.05). Conclusion Bivalent HPV vaccination has an important protective effect on HPV persistent infection, cervical lesions and cervical cancer in high-risk women.

OBJECTIVE:To evaluate the effects of the implementation of National Essential Medicine System on the utilization rate of hormone drugs in primary medical institutions in China systematically.METHODS:Retrieved from CJFD,Wanfang database and VIP,the literatures about the use of hormone drugs were collected before and after the implementation of National Essential Medicine System.Meta-analysis was performed by using Stata 13.0 software after utilization rate dam extraction and quality evaluation with Cochrane system evaluator manual 5.1.0.RESULTS:A total of 20 literatures were included.Results of Meta-analysis showed that utilization rate of hormone drugs after the implementation of National Essential Medicine System was significantly lower than before implementation,with statistical significance [RD =-0.03,95 ％ CI (-0.05,-0.02),P＜ 0.001].Results of subgroup Meta-analysis of utilization rate of hormone drugs in different areas before and after the implementation of National Essential Medicine System showed that the utilization rate of hormone drugs in eastern areas was significantly lower after the implementation of National Essential Medicine System than before [RD=-0.06,95％ CI(0.09,-0.03),P=0.001] with statistical significance.There was no statistical significance in the utilization rate of hormone drugs in middle area [RD=-0.02,95 ％ CI(0.06,0.02),P=0.235] or western area [RD=-0.01,95％ CI (-0.02,0),P=0.122] before and after the implementation of National Essential Medicine System.CONCLUSIONS:The implementation of National Essential Medicine System reduces the utilization of hormone drugs in primary medical institutions.The effect of the eastern area is more obvious than in middle and western area.

Objective To investigate the clinical significance of thyroglobulin antibody (ATG) and thyroid peroxidase antibody (ATPO) in patients with vitiligo. Methods Venous blood samples were obtained from 87 patients with vitiligo and 90 age- and sex-matched normal human controls. Chemiluminescence was applied to measure the serum levels of ATG, ATPO, free triiodothyronine, free tetraiodothyronine and thyroid stimulating hormone (TSH). Results There was a significant increase in the positivity rates of ATG (23.0% vs 6.7%, P < 0.01) and ATPO (24.1% vs 7.8%, P < 0.01) as well as the serum level of TSH (3.4 ± 2.4 vs 2.4 ± 1.2 pmol/L, P < 0.05) in the patients with vitiligo compared with the normal human controls. It is worth mentioning that all patients positive for ATG or ATPO were diagnosed with vitiligo vulgaris. The positivity rates of ATG and ATPO in patients with vitiligo aged from 11 to 20 years and 21 to 40 years were significantly higher than those in age-matched normal controls (all P < 0.05). Also, female patients had a higher positivity rate of ATG and ATPO than female controls did (34.1% vs 8.5%, χ2 = 8.90, P < 0.01; 34.1% vs 10.6%,χ2 = 7.29, P < 0.05). The highest positivity rates of both ATG and ATPO were 53.3%, which were observed in vitiligo patients aged from 11 to 20 years, followed by patients from 21 to 40 years (ATG 34.5%, ATPO 34.5%). In patients with vitiligo positive for both ATG and ATPO, the occurrence of autoimmune thyroid disease was 70% (14/20), significantly higher than that in ATG- and ATPO- positive healthy controls (16.7%, χ2 = 5.4, P < 0.05). Conclusions ATG and ATPO were observed in young female patients with vitiligo vulgaris, and they may be associated with the development of autoimmune thyroid diseases.