Objective:To investigate the influencing factors and perinatal outcomes associated with monozygotic twins (MZT) following elective single embryo transfer (eSET) via in vitro fertilization or intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). Methods:A retrospective cohort study was conducted on 12 079 patients who achieved pregnancy after undergoing IVF/ICSI-eSET at Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University between January 2015 and September 2023. Patients were stratified into two groups based on ultrasound findings 30 d post-transfer: singleton pregnancy group and MZT pregnancy group. Finally, 300 MZT and 1 500 single pregnancies, which were randomly matched according to 1∶5 were included by study period. General patients' characteristics, embryo-related factors, and perinatal outcomes were compared between the two groups. A multivariate logistic regression model was employed to identify risk factors for MZT after single embryo transfer, adjusting for potential confounding variables.Results:The incidence of twin pregnancy following single embryo transfer was 2.48% (300/12 079), which was higher than that of naturally conceived monozygotic twin pregnancy. No significant difference was found in baseline characteristics between the two groups (all P>0.05). The blastocyst transfer rate was higher in the MZT pregnancy group [93.3% (280/300)] than in the singleton pregnancy group [88.8% (1 332/1 500), P=0.022]. Multivariate logistic regression analysis also showed that blastocyst transfer was associated with an increased risk of MZT ( OR=0.552, P=0.016, 95% CI: 0.341-0.894). Analysis of blastocyst cycles showed that the risk of MZT was higher when transferring high-quality blastocysts [79.6% (223/280) vs. 67.8% (903/1 332), P<0.001], where as a trophectoderm (TE) grading of C [20.4% (57/280) vs. 32.2% (429/1 332), P<0.001] had a lower risk of MZT. After adjusting for confounding factors, the risk of MZT was found to increase with the transfer of blastocysts with a B-grade inner cell mass (ICM) ( OR=0.601, P=0.001, 95% CI: 0.442-0.819) and A/B grade TE (grade A: OR=2.951, P<0.001, 95% CI: 1.980-4.399; grade B: OR=1.840, P<0.001, 95% CI: 1.315-2.576). The risk of complications during pregnancy [47.7% (143/300) vs. 19.3% (289/1 500), P<0.001], preterm labor [55.1% (140/254) vs. 7.4% (101/1 368), P<0.001], and the risk of stillbirth [3.7% (11/300) vs. 1.5% (22/1 500), P=0.016] were significantly higher in the MZT pregnancy group than in the singleton pregnancy group. Conclusion:Assisted reproductive technology may contribute to the risk of MZT. Transfer of blastocysts, particularly those with loose ICM arrangement and dense TE arrangement, appears to increase the risk of MZT in patients undergoing eSET.

Objective:To investigate the impact of different luteal phase support protocols on pregnancy outcomes in patients aged ≤35 years undergoing modified natural cycle frozen-thawed embryo transfer (mNC-FET).Methods:A retrospective cohort study was conducted to analyze 2 086 cycles of patients aged ≤35 years who received mNC-FET cycles in Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2018 to December 2020. The cycles were divided into three groups based on luteal phase support protocols used. The patients received a combination of progesterone soft capsule and dydrogesterone in the group A (446 cycles), the patients received dydrogesterone in the group B (439 cycles), and the patients received a combination of progesterone vaginal sustained-release gel and dydrogesterone in the group C (1 201 cycles). The pregnancy and perinatal outcomes were compared between groups A and B, groups C and B after matching the baseline data in a ratio of 1∶1 using the propensity score matching (PSM). The effect of different luteal phase support on live birth rate was analyzed after adjusting for confounding factors affected by univariate and multivariate generalized estimating equation (GEE).Results:After PSM, there were no significant differences between groups A and B, groups C and B in human chorionic gonadotropin positive rate, clinical pregnancy rate, ectopic pregnancy rate, live birth rate in transplant cycle, incidence of low weight, macrosomia, premature delivery rate, pregnancy complication rate and incidence of birth defects (all P>0.05). GEE analysis showed that three different luteal phase support regimens were not associated with live birth rate. Conclusion:In the mNC-FET cycle, patients aged ≤35 years who chose dydrogesterone alone as luteal phase support drug, had no difference in live birth rate and perinatal outcome between progesterone soft capsules or progesterone vaginal sustained-release gel combined with dydrogesterone, but the outcome still needs to be confirmed by large sample prospective studies.

Objective:To investigate the influencing factors and perinatal outcomes associated with monozygotic twins (MZT) following elective single embryo transfer (eSET) via in vitro fertilization or intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). Methods:A retrospective cohort study was conducted on 12 079 patients who achieved pregnancy after undergoing IVF/ICSI-eSET at Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University between January 2015 and September 2023. Patients were stratified into two groups based on ultrasound findings 30 d post-transfer: singleton pregnancy group and MZT pregnancy group. Finally, 300 MZT and 1 500 single pregnancies, which were randomly matched according to 1∶5 were included by study period. General patients' characteristics, embryo-related factors, and perinatal outcomes were compared between the two groups. A multivariate logistic regression model was employed to identify risk factors for MZT after single embryo transfer, adjusting for potential confounding variables.Results:The incidence of twin pregnancy following single embryo transfer was 2.48% (300/12 079), which was higher than that of naturally conceived monozygotic twin pregnancy. No significant difference was found in baseline characteristics between the two groups (all P>0.05). The blastocyst transfer rate was higher in the MZT pregnancy group [93.3% (280/300)] than in the singleton pregnancy group [88.8% (1 332/1 500), P=0.022]. Multivariate logistic regression analysis also showed that blastocyst transfer was associated with an increased risk of MZT ( OR=0.552, P=0.016, 95% CI: 0.341-0.894). Analysis of blastocyst cycles showed that the risk of MZT was higher when transferring high-quality blastocysts [79.6% (223/280) vs. 67.8% (903/1 332), P<0.001], where as a trophectoderm (TE) grading of C [20.4% (57/280) vs. 32.2% (429/1 332), P<0.001] had a lower risk of MZT. After adjusting for confounding factors, the risk of MZT was found to increase with the transfer of blastocysts with a B-grade inner cell mass (ICM) ( OR=0.601, P=0.001, 95% CI: 0.442-0.819) and A/B grade TE (grade A: OR=2.951, P<0.001, 95% CI: 1.980-4.399; grade B: OR=1.840, P<0.001, 95% CI: 1.315-2.576). The risk of complications during pregnancy [47.7% (143/300) vs. 19.3% (289/1 500), P<0.001], preterm labor [55.1% (140/254) vs. 7.4% (101/1 368), P<0.001], and the risk of stillbirth [3.7% (11/300) vs. 1.5% (22/1 500), P=0.016] were significantly higher in the MZT pregnancy group than in the singleton pregnancy group. Conclusion:Assisted reproductive technology may contribute to the risk of MZT. Transfer of blastocysts, particularly those with loose ICM arrangement and dense TE arrangement, appears to increase the risk of MZT in patients undergoing eSET.

Objective:To investigate the impact of different luteal phase support protocols on pregnancy outcomes in patients aged ≤35 years undergoing modified natural cycle frozen-thawed embryo transfer (mNC-FET).Methods:A retrospective cohort study was conducted to analyze 2 086 cycles of patients aged ≤35 years who received mNC-FET cycles in Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2018 to December 2020. The cycles were divided into three groups based on luteal phase support protocols used. The patients received a combination of progesterone soft capsule and dydrogesterone in the group A (446 cycles), the patients received dydrogesterone in the group B (439 cycles), and the patients received a combination of progesterone vaginal sustained-release gel and dydrogesterone in the group C (1 201 cycles). The pregnancy and perinatal outcomes were compared between groups A and B, groups C and B after matching the baseline data in a ratio of 1∶1 using the propensity score matching (PSM). The effect of different luteal phase support on live birth rate was analyzed after adjusting for confounding factors affected by univariate and multivariate generalized estimating equation (GEE).Results:After PSM, there were no significant differences between groups A and B, groups C and B in human chorionic gonadotropin positive rate, clinical pregnancy rate, ectopic pregnancy rate, live birth rate in transplant cycle, incidence of low weight, macrosomia, premature delivery rate, pregnancy complication rate and incidence of birth defects (all P>0.05). GEE analysis showed that three different luteal phase support regimens were not associated with live birth rate. Conclusion:In the mNC-FET cycle, patients aged ≤35 years who chose dydrogesterone alone as luteal phase support drug, had no difference in live birth rate and perinatal outcome between progesterone soft capsules or progesterone vaginal sustained-release gel combined with dydrogesterone, but the outcome still needs to be confirmed by large sample prospective studies.

Objective:To evaluate the efficacy and safety of a Chinese herbal mask in the treatment of melanized-type melasma by stereological measurements.Methods:A randomized, controlled, evaluator- and statistician-blind clinical study was conducted. A total of 116 patients with melasma who met the inclusion and exclusion criteria were selected and equally divided into two groups by using a random number table method, with 58 patients in each group. The treatment group received topical treatment with a Chinese herbal mask once a night, while the control group received topical hydroquinone cream in the morning and evening twice daily. The course of treatment lasted 12 weeks, with follow-ups every 4 weeks, and an additional follow-up was performed 12 weeks after discontinuation of treatment. A facial image analyzer and a reflectance confocal microscopy (RCM) were used to collect images before and after treatment. Melasma area and severity index (MASI) scores and RCM scores served as outcome measures, and stereological measurement methods were employed to compare scores before and after treatment; response rates and recurrence rates were calculated, and adverse reactions were observed and recorded. Statistical analyses were performed using the chi-square test and t test. Results:A total of 54 patients in the treatment group and 47 patients in the control group completed the study. After 4- and 8-week treatment, there were no significant differences in MASI scores or RCM scores between the two groups (all P > 0.05) ; after 12-week treatment, the MASI scores and RCM scores were significantly lower in the treatment group (2.91 ± 1.16, 5.48 ± 1.60, respectively) than in the control group (3.99 ± 2.03, 6.66 ± 1.88, t = -3.32, -3.37, respectively, both P < 0.01). After the end of treatment, there was no significant difference in the response rate between the treatment group (44/54, 81.5%) and the control group (34/47, 72.3%; χ2 = 0.12, P > 0.05). Twelve weeks after discontinuation of treatment, 2 patients (3.7%) relapsed in the treatment group, while 15 (31.9%) relapsed in the control group, showing a significant difference in recurrence rates between the two groups ( χ2 = 12.34, P < 0.01). Adverse reactions occurred in 2 patients (3.7%) in the treatment group and 6 (11.5%) in the control group, and there was no significant difference in the incidence of adverse reactions between the two groups ( χ2 = 1.34, P > 0.05) . Conclusions:The Chinese herbal mask was safe and effective in the treatment of melanized-type melasma. The application of the facial image analyzer and RCM technology in combination with stereological measurement methods has certain value in reducing the subjectivity in efficacy evaluation for melasma.

Objective To exploring the effect of inflammatory indicators on the outcome of assisted conception of in vitro fertilization/intracytoplasmic sperm injection(IVF/ICSI)in patients with ovarian endometriosis(OEM).Methods The study subjects were selected as the patients with OEM who received IVF/ICSI treatment at the Third Affiliated Hospital of Zhengzhou University from January 2019 to February 2024.Based on whether previous surgery for uterine endometriosis cyst removal had been performed,they were divided into non-surgery group(n=73)and surgery group(n=112).To explore the differences in inflammatory markers between two groups and to determine whether they have an impact on the outcome of assisted reproduction.Results The lymphocytes(L)and blastocyst formation rates of patients in surgery group were significantly higher than those in non-surgery group,platelet to lymphocyte ratio(PLR)and carbohydrate antigen 125(CA125)were significantly lower than those in non-surgery group(P＜0.05).There were no statistically significant differences in the clinical pregnancy rate and live birth rate between two groups of patients(P＞0.05).After adjusting for confounding factors,the results of multivariate Logistic regression analysis showed that L,PLR,and CA125 had no statistically significant impact on the clinical pregnancy rate and live birth rate(P＞0.05).Both maternal age and the number of embryos transferred exerted a certain influence on clinical pregnancy rates and live birth rates(P＜0.05),furthermore,the type of embryo transferred had a significant effect on clinical pregnancy rates(P＜0.05).Conclusion Compared with non-surgery group,surgery group had higher L level and lower PRL and CA125 levels,but these changes did not significantly affect clinical pregnancy or live birth outcomes in OEM patients undergoing IVF/ICSI assisted reproduction.

To investigate the immune protective effect induced by superoxide dismutase(SOD)of the Toxocara canis(T.canis)in mice,in this experiment,seventy-five Kunming mice of six-week-age were randomly divided into 5 groups for the immune protection experiment,the mice were subcutaneously immunized with 50,75 and 125 mg/L Tc-SOD(0.1 mL)on days 0,14 and 28,re-spectively.In the blank group,mice did not receive any treatment,while the PBS group was injected with equal amounts of sterility PBS.A total of three immunizations and each immunization interval of 14 days.14 days after the third immunization,the 3 000 infected worm eggs were given orally in all groups.Before each immunization,14 days after the third immunization,and after the T.canis attack on the 7th day,the blood was sampled from the tail vein and sera were separated,while the IgG levels in serum were detected by indirect ELISA.The proliferation ability of splenic lympho-cytes and the mRNA expression levels of Th1(IFN-γ,IL-2)and Th2 cytokines(IL-4,IL-10)were analyzed with CCK-8 and qRT-PCR.The larvae reduction rates were calculated on the 7th day after the T.canis attack,and the pathological changes in the livers and lungs were observed using H&E staining.The results showed that compared with the PBS group,the serum IgG antibody levels increased with the frequency and duration of Tc-SOD immunization.It remained at a high level af-ter the T.canis attack,and the differences were very significant(P＜0.001).The mRNA expres-sion levels of IFN-γ,IL-2,IL-4,and IL-10 were dramatically increased(P＜0.01),while IL-4 andIL-10 were significantly higher than IFN-γ and IL-2 on the 14th day after the third immunization(P＜0.05),showing the Th2 type predominant immune response.And induce the proliferation of spleen lymphocytes in vitro(P＜0.01).The reduction rates of larvae of 50,75 and 125 mg/L Tc-SOD immune groups were 18.7％,24.9％and 37.6％,respectively,with significant differences in the 125 mg/L Tc-SOD group(P＜0.05),and had better immune protection effect.Moreover,the H&E staining results indicated that three Tc-SOD immune groups reduce the inflammatory cell infiltration(neutrophils,eosinophils,and macrophages)and hemorrhagic lesions in the livers and lungs of mice.The above results indicated that Tc-SOD could induce humoral and cellular immune responses in mice,mainly Th2 immune response,and provide immune protection against T.canis infection.

Objective:To evaluate the efficacy and safety of a Chinese herbal mask in the treatment of melanized-type melasma by stereological measurements.Methods:A randomized, controlled, evaluator- and statistician-blind clinical study was conducted. A total of 116 patients with melasma who met the inclusion and exclusion criteria were selected and equally divided into two groups by using a random number table method, with 58 patients in each group. The treatment group received topical treatment with a Chinese herbal mask once a night, while the control group received topical hydroquinone cream in the morning and evening twice daily. The course of treatment lasted 12 weeks, with follow-ups every 4 weeks, and an additional follow-up was performed 12 weeks after discontinuation of treatment. A facial image analyzer and a reflectance confocal microscopy (RCM) were used to collect images before and after treatment. Melasma area and severity index (MASI) scores and RCM scores served as outcome measures, and stereological measurement methods were employed to compare scores before and after treatment; response rates and recurrence rates were calculated, and adverse reactions were observed and recorded. Statistical analyses were performed using the chi-square test and t test. Results:A total of 54 patients in the treatment group and 47 patients in the control group completed the study. After 4- and 8-week treatment, there were no significant differences in MASI scores or RCM scores between the two groups (all P > 0.05) ; after 12-week treatment, the MASI scores and RCM scores were significantly lower in the treatment group (2.91 ± 1.16, 5.48 ± 1.60, respectively) than in the control group (3.99 ± 2.03, 6.66 ± 1.88, t = -3.32, -3.37, respectively, both P < 0.01). After the end of treatment, there was no significant difference in the response rate between the treatment group (44/54, 81.5%) and the control group (34/47, 72.3%; χ2 = 0.12, P > 0.05). Twelve weeks after discontinuation of treatment, 2 patients (3.7%) relapsed in the treatment group, while 15 (31.9%) relapsed in the control group, showing a significant difference in recurrence rates between the two groups ( χ2 = 12.34, P < 0.01). Adverse reactions occurred in 2 patients (3.7%) in the treatment group and 6 (11.5%) in the control group, and there was no significant difference in the incidence of adverse reactions between the two groups ( χ2 = 1.34, P > 0.05) . Conclusions:The Chinese herbal mask was safe and effective in the treatment of melanized-type melasma. The application of the facial image analyzer and RCM technology in combination with stereological measurement methods has certain value in reducing the subjectivity in efficacy evaluation for melasma.

Objective To exploring the effect of inflammatory indicators on the outcome of assisted conception of in vitro fertilization/intracytoplasmic sperm injection(IVF/ICSI)in patients with ovarian endometriosis(OEM).Methods The study subjects were selected as the patients with OEM who received IVF/ICSI treatment at the Third Affiliated Hospital of Zhengzhou University from January 2019 to February 2024.Based on whether previous surgery for uterine endometriosis cyst removal had been performed,they were divided into non-surgery group(n=73)and surgery group(n=112).To explore the differences in inflammatory markers between two groups and to determine whether they have an impact on the outcome of assisted reproduction.Results The lymphocytes(L)and blastocyst formation rates of patients in surgery group were significantly higher than those in non-surgery group,platelet to lymphocyte ratio(PLR)and carbohydrate antigen 125(CA125)were significantly lower than those in non-surgery group(P＜0.05).There were no statistically significant differences in the clinical pregnancy rate and live birth rate between two groups of patients(P＞0.05).After adjusting for confounding factors,the results of multivariate Logistic regression analysis showed that L,PLR,and CA125 had no statistically significant impact on the clinical pregnancy rate and live birth rate(P＞0.05).Both maternal age and the number of embryos transferred exerted a certain influence on clinical pregnancy rates and live birth rates(P＜0.05),furthermore,the type of embryo transferred had a significant effect on clinical pregnancy rates(P＜0.05).Conclusion Compared with non-surgery group,surgery group had higher L level and lower PRL and CA125 levels,but these changes did not significantly affect clinical pregnancy or live birth outcomes in OEM patients undergoing IVF/ICSI assisted reproduction.

To investigate the immune protective effect induced by superoxide dismutase(SOD)of the Toxocara canis(T.canis)in mice,in this experiment,seventy-five Kunming mice of six-week-age were randomly divided into 5 groups for the immune protection experiment,the mice were subcutaneously immunized with 50,75 and 125 mg/L Tc-SOD(0.1 mL)on days 0,14 and 28,re-spectively.In the blank group,mice did not receive any treatment,while the PBS group was injected with equal amounts of sterility PBS.A total of three immunizations and each immunization interval of 14 days.14 days after the third immunization,the 3 000 infected worm eggs were given orally in all groups.Before each immunization,14 days after the third immunization,and after the T.canis attack on the 7th day,the blood was sampled from the tail vein and sera were separated,while the IgG levels in serum were detected by indirect ELISA.The proliferation ability of splenic lympho-cytes and the mRNA expression levels of Th1(IFN-γ,IL-2)and Th2 cytokines(IL-4,IL-10)were analyzed with CCK-8 and qRT-PCR.The larvae reduction rates were calculated on the 7th day after the T.canis attack,and the pathological changes in the livers and lungs were observed using H&E staining.The results showed that compared with the PBS group,the serum IgG antibody levels increased with the frequency and duration of Tc-SOD immunization.It remained at a high level af-ter the T.canis attack,and the differences were very significant(P＜0.001).The mRNA expres-sion levels of IFN-γ,IL-2,IL-4,and IL-10 were dramatically increased(P＜0.01),while IL-4 andIL-10 were significantly higher than IFN-γ and IL-2 on the 14th day after the third immunization(P＜0.05),showing the Th2 type predominant immune response.And induce the proliferation of spleen lymphocytes in vitro(P＜0.01).The reduction rates of larvae of 50,75 and 125 mg/L Tc-SOD immune groups were 18.7％,24.9％and 37.6％,respectively,with significant differences in the 125 mg/L Tc-SOD group(P＜0.05),and had better immune protection effect.Moreover,the H&E staining results indicated that three Tc-SOD immune groups reduce the inflammatory cell infiltration(neutrophils,eosinophils,and macrophages)and hemorrhagic lesions in the livers and lungs of mice.The above results indicated that Tc-SOD could induce humoral and cellular immune responses in mice,mainly Th2 immune response,and provide immune protection against T.canis infection.