ObjectiveTo determine whether Jinshui Huanxian component formula Ⅱ （ECC-JHF Ⅱ） and matrix hardness interfere with the process of epithelial-mesenchymal transition （EMT） during pulmonary fibrosis by regulating integrin β₁ （ITG β₁）， and to elucidate the underlying mechanism. MethodsA pulmonary fibrosis mouse model was established by a single intratracheal instillation of bleomycin （BLM）. To observe the disease progression at various time points， the mice were sacrificed before modeling and on days 7， 14， 21， 28 and 42 after modeling for assessment of relevant indicators. To evaluate the therapeutic effect of drug intervention， the mice were given ECC-JHF Ⅱ and pirfenidone （PFD） by gavage starting on day 29 after modeling for 14 days of treatment， after which the mice were sacrificed. Pulmonary function， pathological changes， collagen deposition， lung tissue hardness and EMT markers were detected. Polyethylene glycol （PEG） composite hydrogels with different hardness were prepared in vitro using biomaterials such as eight-arm polyethylene glycol thiol （PEG-SH） and eight-arm polyethylene glycol maleimide （PEG-MAL）. Human type Ⅱ alveolar epithelial cells cultured in hard matrix culture dishes were treated with low， medium， and high doses of ECC-JHF Ⅱ （15.31， 30.63， 61.25 mg·L^-1） to observe the intervention effect of ECC-JHF Ⅱ on EMT induced by transforming growth factor beta 1 （TGF-β₁）. Human type Ⅱ alveolar epithelial cells cultured on soft （3.8 kPa） and hard （culture dishes， ~ GPa） matrices were treated with ECC-JHF Ⅱ， respectively. The mRNA and fluorescence expression levels of the epithelial marker E-cadherin （CDH1）， the mesenchymal marker N-cadherin （CDH2） and vimentin （VIM）， as well as ITG β₁， were detected by real-time quantitative polymerase chain reaction （Real-time PCR） and immunofluorescence. Western blot was used to detect the protein expression of CDH1， CDH2 and ITG β₁， and the biomechanical mechanism of ECC-JHF Ⅱ on EMT and pulmonary fibrosis was further verified by knocking down and specific activation of ITG β₁. ResultsECC-JHF Ⅱ treatment improved BLM-induced pulmonary fibrosis in mice， as evidenced by significantly increased minute ventilation （MV） and 50% expiratory flow at tidal volume （EF₅₀） （P<0.05）， and significantly increased tidal volume （TV） after treatment （P<0.01） compared with the model group. Immunohistopathological analysis revealed attenuated inflammation and alveolar injury， reduced deposition of type Ⅰ collagen （Col-Ⅰ）， and significantly decreased lung tissue stiffness （P<0.01）， thereby alleviating disease progression. The real-time PCR results showed that， compared with the model group， the ECC-JHF Ⅱ group showed increased CDH1 expression and decreased CDH2 and VIM expression in mouse lung tissues （P<0.05， P<0.01）， indicating that ECC-JHF Ⅱ inhibited the EMT process to ameliorate pulmonary fibrosis. In vitro experimental results showed that both ECC-JHF Ⅱ and matrix softening reversed the EMT phenotype switch. Specifically， compared with the hard matrix group， the cells cultured on the soft matrix exhibited significantly upregulated expression of the epithelial marker CDH1 （P<0.01） and significantly downregulated expression of the mesenchymal markers CDH2 and VIM （P<0.05）. Furthermore， ECC-JHF Ⅱ inhibited the expression of ITG β₁ both in vitro and in vivo. Knockdown and specific activation of ITG β₁ further confirmed that ITG β₁ is involved in the regulatory process by which ECC-JHF Ⅱ inhibits EMT and ameliorates pulmonary fibrosis. ConclusionECC-JHF Ⅱ and matrix softening can block the EMT process and ameliorate pulmonary fibrosis， and the underlying mechanism may be associated with the inhibition of ITG β₁-mediated mechanotransduction. This study reveals a novel mechanism of ECC-JHF Ⅱ intervention in pulmonary fibrosis from a biomechanical perspective.

Objective:To investigate the perinatal outcome of selective fetal reduction of dizygotic twins pregnancies after double embryo transferred assisted by in vitro fertilization/ intracytoplasmic sperm injection (IVF/ICSI). Methods:The clinical data of patients with single birth who underwent IVF/ICSI after double embryo transferred from September 1, 2005 to July 31, 2020 in Reproductive Center of the Third Affiliated Hospital of Zhengzhou University were retrospectively analyzed. The patients were divided into three groups according to the occurrence of fetal reduction: 80 cases of dizygotic twins to singleton by selective fetal reduction (group A), 832 cases of dizygotic twins to singleton by spontaneous fetal reduction (group B), and 6 178 cases of singleton (group C). Using group A as the reference, after approximately propensity score matching (PSM) with 1∶4 ratio, 289 cases in group B and 271 cases in group C were obtained. The differences in baseline data and perinatal outcomes between group A and group B or group C were analyzed.Results:1) Before PSM, there were statistically significant differences in patients' age, previous fertility history, type of infertility, and stage of embryo transferred between group A and group B or group C (all P<0.05); after PSM, there were no statistically significant differences in baseline data between group A and group B or group C (all P>0.05). 2) After PSM, the premature birth rate was higher in group A than in group B [17.5% (14/80) vs. 8.7% (25/289), P=0.023] and the gestational weeks [(38.18±1.98) weeks vs. (38.64±1.83) weeks, P=0.034] were lower; group A had higher incidence rates of premature birth [17.5% (14/80) vs. 7.7% (21/271), P=0.011] and premature rupture of membranes [5.0% (4/80) vs. 0.4% (1/271), P=0.002] than those in group C, but gestational weeks [(38.18±1.98) weeks vs. (38.85±1.47) weeks, P=0.002] and birth weight [(3 253.07±475.73) g vs. (3 384.89±479.54) g, P=0.029] were lower. Conclusion:Perinatal outcomes of selective fetal reduction of dizygotic twins pregnancies after assisted by IVF/ICSI were poor. Selective fetal reduction in multiple pregnancies is not the best remedy. The strategy of selective single embryo transfer should be adopted to effectively reduce the rate of multiple pregnancies and improve maternal and infant outcomes.

Objective:To investigate the perinatal outcome of selective fetal reduction of dizygotic twins pregnancies after double embryo transferred assisted by in vitro fertilization/ intracytoplasmic sperm injection (IVF/ICSI). Methods:The clinical data of patients with single birth who underwent IVF/ICSI after double embryo transferred from September 1, 2005 to July 31, 2020 in Reproductive Center of the Third Affiliated Hospital of Zhengzhou University were retrospectively analyzed. The patients were divided into three groups according to the occurrence of fetal reduction: 80 cases of dizygotic twins to singleton by selective fetal reduction (group A), 832 cases of dizygotic twins to singleton by spontaneous fetal reduction (group B), and 6 178 cases of singleton (group C). Using group A as the reference, after approximately propensity score matching (PSM) with 1∶4 ratio, 289 cases in group B and 271 cases in group C were obtained. The differences in baseline data and perinatal outcomes between group A and group B or group C were analyzed.Results:1) Before PSM, there were statistically significant differences in patients' age, previous fertility history, type of infertility, and stage of embryo transferred between group A and group B or group C (all P<0.05); after PSM, there were no statistically significant differences in baseline data between group A and group B or group C (all P>0.05). 2) After PSM, the premature birth rate was higher in group A than in group B [17.5% (14/80) vs. 8.7% (25/289), P=0.023] and the gestational weeks [(38.18±1.98) weeks vs. (38.64±1.83) weeks, P=0.034] were lower; group A had higher incidence rates of premature birth [17.5% (14/80) vs. 7.7% (21/271), P=0.011] and premature rupture of membranes [5.0% (4/80) vs. 0.4% (1/271), P=0.002] than those in group C, but gestational weeks [(38.18±1.98) weeks vs. (38.85±1.47) weeks, P=0.002] and birth weight [(3 253.07±475.73) g vs. (3 384.89±479.54) g, P=0.029] were lower. Conclusion:Perinatal outcomes of selective fetal reduction of dizygotic twins pregnancies after assisted by IVF/ICSI were poor. Selective fetal reduction in multiple pregnancies is not the best remedy. The strategy of selective single embryo transfer should be adopted to effectively reduce the rate of multiple pregnancies and improve maternal and infant outcomes.

Objective:To investigate the obstetric outcomes of intracytoplasmic sperm injection and frozen-thawed embryo transfer (ICSI-FET) between donor and autologous oocytes.Methods:A retrospective cohort study was conducted to analyze the clinical data of pregnant patients who underwent ICSI-FET in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from June 2016 to January 2020. Totally 73 patients with donor oocytes, and 550 patients who used autologous oocytes in the same period, and the patients were matched at 1∶3 with propensity score matching (PSM), then there were 47 patients in donor group and 131 patients in autologous group. The general conditions and obstetric outcomes were compared among donor group and autologous group. Multivariate logistic regression and linear regression were applied to analyze the factors affecting perinatal complications.Results:The bilateral antral follicle count (3.08±4.78) and basal estradiol level [(71.55±45.29) pmol/L] in donor group were significantly lower than those in autologous group [14.95±6.42, (132.84±74.89) pmol/L, all P<0.001]. The birth weight of singleton in donor group [(2 916.48±537.55) g] was lower than that in autologous group [(3 326.67±503.43) g], and there was significant difference ( P<0.001). There were no significant differences in premature birth rate [21.28% (10/47) vs. 16.03% (21/131), P=0.416] and incidence of hypertensive disorder complicating pregnancy [12.77% (6/47) vs. 7.63% (10/131), P=0.448] between donor group and autologous group, but both of them had an increasing trend in donor group. Oocyte-donated ICSI-FET reduced the birth weight of singleton (MD=-388.225, 95% CI=-625.914--150.537, P=0.002). Conclusion:The perinatal outcome of oocyte-donated ICSI-FET is relatively safe, but the birth weight of singleton is lower than that of self-oocyte ICSI-FET.

Objective:To investigate the obstetric outcomes of intracytoplasmic sperm injection and frozen-thawed embryo transfer (ICSI-FET) between donor and autologous oocytes.Methods:A retrospective cohort study was conducted to analyze the clinical data of pregnant patients who underwent ICSI-FET in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from June 2016 to January 2020. Totally 73 patients with donor oocytes, and 550 patients who used autologous oocytes in the same period, and the patients were matched at 1∶3 with propensity score matching (PSM), then there were 47 patients in donor group and 131 patients in autologous group. The general conditions and obstetric outcomes were compared among donor group and autologous group. Multivariate logistic regression and linear regression were applied to analyze the factors affecting perinatal complications.Results:The bilateral antral follicle count (3.08±4.78) and basal estradiol level [(71.55±45.29) pmol/L] in donor group were significantly lower than those in autologous group [14.95±6.42, (132.84±74.89) pmol/L, all P<0.001]. The birth weight of singleton in donor group [(2 916.48±537.55) g] was lower than that in autologous group [(3 326.67±503.43) g], and there was significant difference ( P<0.001). There were no significant differences in premature birth rate [21.28% (10/47) vs. 16.03% (21/131), P=0.416] and incidence of hypertensive disorder complicating pregnancy [12.77% (6/47) vs. 7.63% (10/131), P=0.448] between donor group and autologous group, but both of them had an increasing trend in donor group. Oocyte-donated ICSI-FET reduced the birth weight of singleton (MD=-388.225, 95% CI=-625.914--150.537, P=0.002). Conclusion:The perinatal outcome of oocyte-donated ICSI-FET is relatively safe, but the birth weight of singleton is lower than that of self-oocyte ICSI-FET.