Objective: To examine the causal relationship between inflammatory factors and breast cancer using two-sample Mendelian randomization (MR) approach, so as to provide the basis for the prevention and treatment of breast cancer. Methods: Data of 91 inflammatory cytokines (n=14 824) and 5 subtypes of breast cancer (n=247 173) were collected from genome-wide association studies (GWAS). Single nucleotide polymorphism (SNP) associated with 91 inflammatory factors were selected as instrumental variables. MR analyses were performed using the inverse-variance weighted (IVW) method with inflammatory factors as exposure factors and breast cancer as outcome variables. The risk of type I error and the effect of multiple testing were reduced using the FDR correction method. The stability and reliability of the results were verified using Steiger test of directionality, MR-Egger regression, MR-PRESSO test and leave-one out method. Results: Twenty-three inflammatory factors, including β nerve growth factor, interleukin-5, cystatin D and C-X-C chemokine ligand 1 were statistically associated with breast cancer (all P<0.05). After FDR adjustment, only evaluated abundance of oncostatin-M was found to be statistically associated with an increased risk of Basal-like (triple-negative) breast cancer (OR=1.186, 95%CI: 1.081-1.302, P=0.001, q=0.029), and the other 22 inflammatory factors had a high risk of type I error (all q>0.1). The sensitivity analysis indicated that the results were robust. No instrumental variables were found to have a significant impact on the results, which could exclude the influence of heterogeneity, horizontal pleiotropy, and reverse causality on the outcome. Conclusion The increased abundance of oncostatin-M may increase the risk of Basal-like (triple-negative) breast cancer.

Objective:To discuss the effect of Wnt/β-catenin signaling pathway inhibitor methyl 3-｛[(4-methyl-phenyl)sulfonyl]amino ｝ benzoate(MS AB)on the fibrogenic response of the human endometrial stromal cells(HESCs),and to provide the foundation for the application of MSAB in the target therapy of intrauteriue adhesion(JUA).Methods:The normal HESCs were cultured in vitro and divided into two groups:control group and transforming growth factor β1(TGF-β1)group;the HESCs from the adhesion part of the IUA patients were cultured in vitro,regarded as IUA group.Western blotting method was used to detect the expression levels of fibrotic marker protein type Ⅰ collagen α1(COL1A1)in the cells in various groups at different time points(0,12,24,48,and 60 h)after treated with TGF-β1.MTT assay was used to detect the proliferation activities of the cells in various groups.Western blotting method was used to detect the expression levels of the fibrotic marker protein COL1A1,stromal marker proteins such as N-cadherin and α-smooth muscle actin(α-SMA),and Wnt/β-catenin signaling pathway-related protein β-catenin in the cells in control and IUA groups.Based on the MSAB concentrations,the normal HESCs were divided into 0(control),0.25,0.50,0.75,and 1.00 μmol·L-1 MSAB groups,and MTT assay was used to detect the survival rates of the cells in various groups.After treated with MSAB,the normal HESCs were divided into control group(normal HESCs),TGF-β1 group(10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn,replaced with complete culture medium,and the cells continued to be cultured for 24 h),and MSAB group(10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn,replaced with a complete medium containing 0.75 μmol·L-1 MSAB and the cells continued to be cultured for 24 h).Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of epithelial-mesenchymal transition(EMT)-related transcription factors Snail,Slug,Smuc,ZEB1,and ZEB2,and COL1A1 mRNA in the cells in various groups.Western blotting method was used to detect the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in various groups.Results:Compared with control group(after treated with TGF-β1 for 0 h),the expression levels of COL1A1 proteins in the HESCs after treated with TGF-β1 for 12,24,48,and 60 h in TGF-β1 group were increased(P＜0.05 or P＜0.01).Compared with control group,there was no significant difference in the proliferation activity of the HESCs in IUA and TGF-β1 groups(P＞0.05).Compared with control group,the expression levels of COL1A1,β-catenin,N-cadherin,and α-SMA proteins in the cells in IUA group were increased(P＜0.05 or P＜0.01).Compared with control group,the survival rates of the cells in 0.75 and 1.00 μmol·L-1 MSAB groups were decreased(P＜0.05 or P＜0.01).Compared with control group,the expression levels of Snail,Slug,and COL1A1 mRNA in the cells in TGF-β1 group were increased(P＜0.05 or P＜0.01);compared with TGF-β1 group,the expression levels of Snail,Slug,and COL1A1 mRNA in the cells in MSAB group were decreased(P＜0.05 or P＜0.01).Compared with control group,after treated with TGF-β1 for 24 h,the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in TGF-β1 group were increased(P＜0.01);compared with TGF-β1 group,the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in MSAB group were decreased(P＜0.05 or P＜0.01).Conclusion:MSAB can inhibit the fibrogenic responses of the HESCs in vitro,and the results provide the theoretical basis for the application of MSAB in the target therapy of IUA.

Objective:To investigate the effects of bisphenol A(BPA)on the proliferation activity and stemness characteristics of endometrial mesenchymal stem/stromal cells(eMSCs),and to elucidate the improvement effect of human umbilical cord mesenchymal stem cell-derived supernatant(hUCMSC-Sup)on the cell injury.Methods:The eMSCs were cultured in vitro and treated with different concentrations of BPA(0,200,250,300,350,and 400 μmol·L-1).The eMSCs were divided into control group(only cultured with culture solution),BPA group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA),BPA+hUCMSC-Sup group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA and 50%volumetric ratio of hUCMSC-Sup),and BPA+CHIR-99021 group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA and 10 μmol·L-1 CHIR-99021).The survival rates of eMSCs in various groups were detected by methyl thiazolyl tetrazolium(MTT)assay.The numbers and diameters of the spheroids in various groups were detected by spheroids formation assay,the proliferation activities of the cells in eMSCs stem cell spheroids in various groups were detected by CCK-8 assay;the percentage of CD73+cells in eMSCs in various groups were detected by flow cytometry;the expression levels of sex determining region Y-box 2(Sox2),octamer-binding transcription factor 4(Oct4),and Nanog mRNA in the eMSCs in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method,the expression levels of β-catenin protein in the eMSCs in various groups were detected by Western blotting method.Results:The MTT results showed that after treated with BPA for 24 and 48 h,compared with 0 μmol·L-1 BPA group,the survival rates of eMSCs in 200,250,300,350,and 400 μmol·L-1 BPA groups were significantly decreased(P<0.01).At 24 and 48 h after treatment,compared with control group,the survival rate of the eMSCs in BPA group was significantly decreased(P<0.01);at 48 h after treatment,compared with BPA group,the survival rate of the eMSCs in BPA+hUCMSC-Sup group was significantly inereased(P<0.05).The spheroids formation assay results showed that compared with culture 3 d group,the numbers and diameters of stem cell spheroids of the eMSCs in culture 4 d group and culture 5 d group were significantly increased(P<0.05 or P<0.01);compared with control group,after 48 h of culture,the number and diameter of the cells in eMSCs stem cell spheroids in BPA group were significantly decreased(P<0.05 or P<0.01).The CCK-8 results showed that after 24 and 48 h of treatment,compared with control group,the proliferation activity of the cells in eMSCs stem cell spheroids in BPA group was significantly decreased(P<0.01);compared with BPA group,the proliferation activity of the cells in eMSCs stem cell spheroids in BPA+hUCMSC-Sup group was significantly increased(P<0.01).The flow cytometry results showed that compared with control group,the percentage of the CD73+cells in the eMSCs in BPA group was significantly decreased(P<0.01);compared with BPA group,the percentage of the CD73+cells in eMSCs in BPA+hUCMSC-Sup group was significantly increased(P<0.01).The RT-qPCR results showed that compared with control group,the expression levels of Sox2,Oct4,and Nanog mRNA in the cells in BPA group were significantly decreased(P<0.01);compared with BPA group,the expression levels of Sox2,Oct4,and Nanog mRNA in the cells in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were significantly increased(P<0.01).The Western blotting results showed that compared with control group,the expression level of β-catenin protein in the eMSCs in BPA group was significantly decreased(P<0.01);compared with BPA group,the expression levels of β-catenin protein in the eMSCs in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were signifrcantly inereased(P<0.01).Conclusion:BPA can inhibit the stemness characteristics of the eMSCs,and injury the self-renewal and repair of endometrium;its mechanism may be related to down-regulating the activity of Wnt/β-catenin signal pathway in the cells.hUCMSC-Sup can promote the proliferation of injured eMSCs,and has improvement effect on the stemness injury induced by BPA.

Allergic inflammation is closely related to the activation of mast cells (MCs), which is regulated by its intracellular Ca²⁺ level, but the intake and effects of the intracellular Ca²⁺ remain unclear. The Ca²⁺ influx is controlled by members of Ca²⁺ channels, among which calcium voltage-gated channel subunit alpha1 C (Ca_V1.2) is the most robust. This study aimed to reveal the role and underlying mechanism of MC Ca_V1.2 in allergic inflammation. We found that Ca_V1.2 participated in MC activation and allergic inflammation. Nimodipine (Nim), as a strong Ca_V1.2-specific antagonist, ameliorated allergic inflammation in mice. Further, Ca_V1.2 activation in MC was triggered by phosphatizing at its Ser1928 through protein kinase C (PKC), which calcium/calmodulin-dependent protein kinase II (CaMKII) catalyzed. Overexpression or knockdown of MC Ca_V1.2 influenced MC activation. Importantly, Ca_V1.2 expression in MC had detrimental effects, while its deficiency ameliorated allergic pulmonary inflammation. Results provide novel insights into Ca_V1.2 function and a potential drug target for controlling allergic inflammation.

Allergic inflammation is closely related to the activation of mast cells(MCs),which is regulated by its intracellular Ca2+level,but the intake and effects of the intracellular Ca2+remain unclear.The Ca2+influx is controlled by members of Ca2+channels,among which calcium voltage-gated channel subunit alpha1 C(Cav1.2)is the most robust.This study aimed to reveal the role and underlying mechanism of MC Cav1.2 in allergic inflammation.We found that Cav1.2 participated in MC activation and allergic inflammation.Nimodipine(Nim),as a strong Cav1.2-specific antagonist,ameliorated allergic inflammation in mice.Further,Cav1.2 activation in MC was triggered by phosphatizing at its Ser1928 through protein kinase C(PKC),which calcium/calmodulin-dependent protein kinase Ⅱ(CaMKⅡ)catalyzed.Overexpression or knockdown of MC Cav1.2 influenced MC activation.Importantly,Cav1.2 expression in MC had detrimental effects,while its deficiency ameliorated allergic pulmonary inflammation.Results provide novel insights into Cav1.2 function and a potential drug target for controlling allergic inflammation.

Objective: To explore the possible mechanism of moxibustion in myocardial protection of rats undergoing long-term fatigue exercise based on observing the classical pyroptosis pathway mediated by nuclear factor kappa-B (NF-κB)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/cysteinyl aspartate specific proteinase 1 (Caspase-1).Methods: A total of 50 specific-pathogen-free male Sprague-Dawley rats were bought. Ten unqualified rats were excluded, and the remaining 40 rats were divided into a normal group, a normal + Shenque (CV8) group, a model group, a model + non-meridian non-point group, and a model + Shenque (CV8) group according to the random number table method, with 8 rats in each group. Except for rats in the normal group and the normal + Shenque (CV8) group, rats in the other three groups were trained with an incline running table exercise protocol to create a long-term fatigue exercise model, 1 h/time, once a day for 5 d with 2 d off, for a total of 8 weeks. Rats in the normal group received no modeling or intervention. Rats in the normal + Shenque (CV8) group were not modeled but received mild moxibustion at Shenque (CV8); those in the model group were modeled only without intervention; those in the model + non-meridian non-point group received moxibustion at non-meridian and non-point spots after the modeling; those in the model + Shenque (CV8) group received moxibustion at Shenque (CV8) after modeling. The above moxibustion interventions were performed for 15 min/time once daily, for 5 d with 2 d off per week and a total of 8 weeks. Blood was collected from the femoral artery 4 h after the last exercise, and the serum interleukin (IL)-1β and IL-18 levels were measured. The NF-κB, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, and gasdermin D (GSDMD) expression levels were detected by Western blotting. Myocardial morphology and pyroptosis were observed by hematoxylin-eosin (HE) staining and electron microscopy. Results: The HE staining results showed that the myocardial cells in the model group and the model + non-meridian non-point group were disorganized with blurred transverse lines, widened interstitial spaces, interstitial edema, and inflammatory cell infiltration. The structure of myocardial cells in the model + Shenque (CV8) group was clearly visible, with slightly widened interstitial spaces and occasional infiltration of inflammatory cells in the interstitium. Compared with the normal group, the serum IL-1β and IL-18 levels were increased, and myocardial NF-κB, NLRP3, ASC, Caspase-1, and GSDMD expression levels were elevated in the model group and the model + non-meridian non-point group (P<0.01). Compared with the model group, the above indicators did not change significantly in the model + non-meridian non-point group, while all the above indicators were decreased in the model + Shenque (CV8) group (P<0.01). Compared with the model + non-meridian non-point group, all the above biochemical indicators were decreased in the model + Shenque (CV8) group (P<0.01). Transmission electron microscopy showed that the mitochondria number was increased in the model group and the model + non-meridian non-point group, some of the mitochondrial lumen was irregularly enlarged, the cell membrane structure was unclear, and chromatin was aggregated. The mitochondria number was increased, the swelling was reduced, and the nuclear membrane structure was more intact in the model + Shenque (CV8) group. Conclusion: Moxibustion at Shenque (CV8) regulates the NF-κB/NLRP3/Caspase-1 pathway and reduces the pyroptosisin the myocardium of rats with long-term fatigue exercise, thus reducing the myocardial injury caused by long-term fatigue exercise.

Objective:To analyze the factors influencing clinical pregnancy outcomes after single embryo transfer (SET) undergoing in vitro fertilization-embryo transfer (IVF-ET) treatment cycles. Methods:A retrospective cohort analysis was carried out on SET cycles of Center for Reproductive Medicine in Nanfang Hospital from September 1st, 2013 to December 31st, 2019. Totally 2734 SET cycles were assigned to day 3 (D3) group, day 4 (D4) group, day 5 (D5) group, day 6 (D6) group according to the different stages of embryo development and analyzed for the relation of clinical pregnancy outcomes to ages, different embryo stages and quality.Results:The total clinical pregnancy rate (CPR) was 39.8% (1098/2734) and the live birth rate (LBR) was 30.5% (842/2734) in the 2761 SET cycles. The significant differences were observed among four groups in CPR [D3 group, D4 group, D5 group and D6 group: 33.3% (264/793), 36.4% (142/390), 52.5% (492/937) and 32.6% (200/614), respectively, P<0.001] and LBR [24.8% (203/793), 30.5% (119/390), 40.9% (383/937) and 22.3% (137/614), respectively, P<0.001]. In D3 group, significantly higher CPR and LBR were observed after transfer of top-quality cleavage embryos (8-cell I, 7-cell I, 8-cell II) than those in other cleavage embryos [CPR: 41.7% (207/496) vs. 19.2% (57/297), P<0.001; LBP: 32.1% (159/496) vs. 14.8% (441/297), P<0.001]. In D4 group, significantly higher CPR and LBR were observed after transfer of embryo with entirely compaction than partial compaction [CPR: 40.4% (134/332) vs. 13.8% (8/58), P<0.001; LBR: 34.0% (113/332) vs. 10.3% (113/332), P=0.001]. In slow-growing blastocysts group, fresh transfer of embryos which began blastulation but did not reach Gardner stage III by D5 resulted in similar outcomes to the transfer of fully expanded blastocysts by D6 [CPR: 30.6% (22/72) vs. 32.6% (200/614), P>0.05; LBR: 27.8% (20/72) vs. 22.3% (137/614), P>0.05]. Conclusion:SET of a top-quality D5 blastocyst or D4 morula can reduce the incidence of multiple pregnancies and obtain the best pregnancy outcome. For slow-growing D5 blastocysts, it may be a strategy to improve pregnancy outcome to continue culture until D6 fully expanded blastocysts and then perform subsequent frozen-thawed embryo transfer. For cleavage embryo, SET of top-quality cleavage embryos (8-cell I, 7-cell I, 8-cell II) also achieved satisfactory pregnancy outcome.

Objective:To analyze the factors influencing clinical pregnancy outcomes after single embryo transfer (SET) undergoing in vitro fertilization-embryo transfer (IVF-ET) treatment cycles. Methods:A retrospective cohort analysis was carried out on SET cycles of Center for Reproductive Medicine in Nanfang Hospital from September 1st, 2013 to December 31st, 2019. Totally 2734 SET cycles were assigned to day 3 (D3) group, day 4 (D4) group, day 5 (D5) group, day 6 (D6) group according to the different stages of embryo development and analyzed for the relation of clinical pregnancy outcomes to ages, different embryo stages and quality.Results:The total clinical pregnancy rate (CPR) was 39.8% (1098/2734) and the live birth rate (LBR) was 30.5% (842/2734) in the 2761 SET cycles. The significant differences were observed among four groups in CPR [D3 group, D4 group, D5 group and D6 group: 33.3% (264/793), 36.4% (142/390), 52.5% (492/937) and 32.6% (200/614), respectively, P<0.001] and LBR [24.8% (203/793), 30.5% (119/390), 40.9% (383/937) and 22.3% (137/614), respectively, P<0.001]. In D3 group, significantly higher CPR and LBR were observed after transfer of top-quality cleavage embryos (8-cell I, 7-cell I, 8-cell II) than those in other cleavage embryos [CPR: 41.7% (207/496) vs. 19.2% (57/297), P<0.001; LBP: 32.1% (159/496) vs. 14.8% (441/297), P<0.001]. In D4 group, significantly higher CPR and LBR were observed after transfer of embryo with entirely compaction than partial compaction [CPR: 40.4% (134/332) vs. 13.8% (8/58), P<0.001; LBR: 34.0% (113/332) vs. 10.3% (113/332), P=0.001]. In slow-growing blastocysts group, fresh transfer of embryos which began blastulation but did not reach Gardner stage III by D5 resulted in similar outcomes to the transfer of fully expanded blastocysts by D6 [CPR: 30.6% (22/72) vs. 32.6% (200/614), P>0.05; LBR: 27.8% (20/72) vs. 22.3% (137/614), P>0.05]. Conclusion:SET of a top-quality D5 blastocyst or D4 morula can reduce the incidence of multiple pregnancies and obtain the best pregnancy outcome. For slow-growing D5 blastocysts, it may be a strategy to improve pregnancy outcome to continue culture until D6 fully expanded blastocysts and then perform subsequent frozen-thawed embryo transfer. For cleavage embryo, SET of top-quality cleavage embryos (8-cell I, 7-cell I, 8-cell II) also achieved satisfactory pregnancy outcome.

Objective To analyze the clinical characteristics in children with digestive tract poison-ing in emergency department and discuss the measures of prevention and treatment. Methods Four hundred and seventy-three cases with digestive tract poisoning who visited the emergency department of Zhengzhou Children′s Hospital were retrospectively analyzed from January 2015 to December 2017. The ages,toxic poi-soning causes,types,clinical features, laboratory examinations, hospitalization expenses and outcomes were analyzed. They were divided into drug poisoning and non-drug poisoning group to compare. Results There were 317 (67. 0%) cases aged 6 days to 3 years old,133 (28. 1%) cases aged 3 to 6 years old,23 cases (4. 9%) >6 years old. The incidence rate was similar in each season but slightly lower in winter. There were 462 (97. 7%) cases of accidental poisoning,of which 377 (79. 7%) cases were mistreated and 85 (18. 0%) cases were mistakenly fed by parents,other 11 (2. 3%) cases were non-accidental poisoning. Three hundreds and thirty-six (71. 0%) cases were drug poisoning,and 137 (29. 0%) cases were non-drug poisoning. Drug poisonings were higher in urban children than in rural children, the difference was statistically significant (χ2 ＝7. 037,P＝0. 008). The percentage of digestive symptoms and blood system symptoms in non-drug poi-soning group were higher than those in drug poisoning group,and the differences were statistically significant (54. 0% vs. 8. 3%,χ2 ＝120. 067,P<0. 001;7. 3% vs. 3. 0%,χ2 ＝4. 491,P ＝0. 034). The percentage of cardiovascular system symptoms and respiratory symptoms in the non-drug poisoning group were lower than that in the drug poisoning group,and the differences were statistically significant (1. 5% vs. 14. 9%, χ2 ＝17. 915,P<0. 001;2. 9% vs. 11. 0%,χ2 ＝8. 050,P＝0. 005). Except for liver function and myocardial en-zyme,the percentage of abnormal laboratory indicators(white blood cells,platelets,blood glucose,lactic acid, electrolyte,coagulation) in non-drug poisoning group were higher than those in the drug poisoning group,and the differences were all statistically significant(all P<0. 05). The hospitalization cost of the non-drug poison-ing group was greater than that of the drug poisoning group,and the difference was statistically significant (Z＝ －12. 444,P<0. 001). Both in the drug-poisoning group and non-drug poisoning group,the cure or im-provement rate of the <6 h treating group were higher than that of the >6h treating group,and the difference was all statistically significant(all P<0. 05). Conclusion Children with acute gastrointestinal poisoning are mostly infants and preschoolers,mainly accidental poisoning,and often taken by mistake. Drug poisonings are mostly found in cities and non-drug poisonings in rural areas. Non-drug poisoning children have more serious damages and higher hospitalization costs than drug poisoning children. Early treatment after poisoning is an important factor to improve cure rate.

OBJECTIVETo use combined G-banding and array-comparative genomic hybridization (aCGH) for the prenatal diagnosis of a fetus with 5q35 deletion syndrome.

METHODSChromosomal karotypes of the fetus and parents were analyzed with G-banding analysis. aCGH was performed to detect minor chromosomal structural abnormalities.

RESULTSThe karyotype of the fetus was ascertained as 46, XY, t(5;10)(q35;p13), and the karyotypes of the parents were normal. aCGH has identified a de novo 1.68 Mb deletion at 5q35.2q35.3 and a 1.44 Mb duplication at 10p14p13.

CONCLUSIONaCGH has a higher resolution and greater accuracy for mapping chromosomal aberrations and is a useful supplement for G banding karyptyping analysis.

Adult ; Chromosome Banding ; Chromosome Deletion ; Chromosomes, Human, Pair 5 ; genetics ; Comparative Genomic Hybridization ; Cri-du-Chat Syndrome ; diagnosis ; embryology ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Karyotyping ; Male ; Prenatal Diagnosis ; Trisomy ; diagnosis ; genetics