Objective:To explore the effectiveness, safety, and cost among urinary follicle-stimulating hormone (u-FSH), recombinant FSH (r-FSH)α, and r-FSHβ in the early follicular phase prolonged protocol for patients under 35 years old with normal ovarian function.Methods:It was a retrospective cohort study. Patients under 35 years old with normal ovarian function who underwent early follicular phase prolonged protocol for ovulation stimulation and using in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) for fertilization in Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2018 to December 2023 were recruited, including the fresh and frozen-thawed embryo transfer (FET) cycles. Patients were divided into u-FSH group, r-FSHα group, and r-FSHβ group. A total of 1 048 ovarian stimulation cycles were included, with 150 cycles, 490 cycles and 408 cycles in the three groups respectively. A total of 710 FET cycles with fresh cycle cancellation were included, with 95 cycles, 320 cycles and 295 cycles in the three groups respectively. The baseline data, pregnancy outcomes, safety, and cost were compared among the three groups. The main observation indicators were cumulative pregnancy rate and cumulative live birth rate (CLBR). A binary logistic regression model was used to control confounding factors, and to analyze the relationship between three ovulation inducing medicine and CLBR. Results:The difference in the number of oocytes retrieved among the u-FSH group, r-FSHα group, and r-FSHβ group was statistically significant [13.0 (10.0, 16.0), 14.0 (11.0, 18.0), 15.0 (11.0, 19.0), respectively, P=0.012], and the difference in the number of 2PN embryos was statistically significant [9.0 (6.0, 12.0), 10.0 (7.0, 13.0), 10.0 (7.0, 13.0), respectively, P=0.046]. There were no statistically significant differences in the number of available embryos, available embryo rate, the number of high-quality embryos, high-quality embryo rate, available blastocyst formation rate, fresh cycle clinical pregnancy rate, live birth rate in fresh cycle, cumulative pregnancy rate of frozen embryos with fresh cycle cancellation, CLBR of frozen embryos with fresh cycle cancellation, cumulative clinical pregnancy rate, CLBR, moderate to severe ovarian hyperstimulation syndrome incidence, ectopic pregnancy rate, multiple pregnancy rate and neonatal malformation rate among the three groups (all P>0.05). In terms of economy, the u-FSH group had the lowest total gonadotropin cost for each patient, while the r-FSHα group had the highest. The differences among the three groups were statistically significant [u-FSH group 4 429.08 (3 198.78, 5 044.23) yuan, r-FSHα group 6 023.72 (5 433.75, 7 529.65) yuan, r-FSHβ group 5 480.00 (4 550.90, 6 437.86) yuan, P<0.001]. Binary logistic regression analysis was conducted, using u-FSH as a control. The CLBR of the r-FSHα group and r-FSHβ group showed no statistically significant difference compared with the u-FSH group (a OR=0.95, 95% CI: 0.57-1.58, P=0.838; a OR=0.89, 95% CI: 0.54-1.48, P=0.654). Conclusion:For patients under 35 years old with normal ovarian function undergoing long protocol ovarian stimulation, the effectiveness and safety of the three ovarian-stimulating medicine are similar, but u-FSH has economic advantages.

Objective:To explore the effectiveness, safety, and cost among urinary follicle-stimulating hormone (u-FSH), recombinant FSH (r-FSH)α, and r-FSHβ in the early follicular phase prolonged protocol for patients under 35 years old with normal ovarian function.Methods:It was a retrospective cohort study. Patients under 35 years old with normal ovarian function who underwent early follicular phase prolonged protocol for ovulation stimulation and using in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) for fertilization in Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2018 to December 2023 were recruited, including the fresh and frozen-thawed embryo transfer (FET) cycles. Patients were divided into u-FSH group, r-FSHα group, and r-FSHβ group. A total of 1 048 ovarian stimulation cycles were included, with 150 cycles, 490 cycles and 408 cycles in the three groups respectively. A total of 710 FET cycles with fresh cycle cancellation were included, with 95 cycles, 320 cycles and 295 cycles in the three groups respectively. The baseline data, pregnancy outcomes, safety, and cost were compared among the three groups. The main observation indicators were cumulative pregnancy rate and cumulative live birth rate (CLBR). A binary logistic regression model was used to control confounding factors, and to analyze the relationship between three ovulation inducing medicine and CLBR. Results:The difference in the number of oocytes retrieved among the u-FSH group, r-FSHα group, and r-FSHβ group was statistically significant [13.0 (10.0, 16.0), 14.0 (11.0, 18.0), 15.0 (11.0, 19.0), respectively, P=0.012], and the difference in the number of 2PN embryos was statistically significant [9.0 (6.0, 12.0), 10.0 (7.0, 13.0), 10.0 (7.0, 13.0), respectively, P=0.046]. There were no statistically significant differences in the number of available embryos, available embryo rate, the number of high-quality embryos, high-quality embryo rate, available blastocyst formation rate, fresh cycle clinical pregnancy rate, live birth rate in fresh cycle, cumulative pregnancy rate of frozen embryos with fresh cycle cancellation, CLBR of frozen embryos with fresh cycle cancellation, cumulative clinical pregnancy rate, CLBR, moderate to severe ovarian hyperstimulation syndrome incidence, ectopic pregnancy rate, multiple pregnancy rate and neonatal malformation rate among the three groups (all P>0.05). In terms of economy, the u-FSH group had the lowest total gonadotropin cost for each patient, while the r-FSHα group had the highest. The differences among the three groups were statistically significant [u-FSH group 4 429.08 (3 198.78, 5 044.23) yuan, r-FSHα group 6 023.72 (5 433.75, 7 529.65) yuan, r-FSHβ group 5 480.00 (4 550.90, 6 437.86) yuan, P<0.001]. Binary logistic regression analysis was conducted, using u-FSH as a control. The CLBR of the r-FSHα group and r-FSHβ group showed no statistically significant difference compared with the u-FSH group (a OR=0.95, 95% CI: 0.57-1.58, P=0.838; a OR=0.89, 95% CI: 0.54-1.48, P=0.654). Conclusion:For patients under 35 years old with normal ovarian function undergoing long protocol ovarian stimulation, the effectiveness and safety of the three ovarian-stimulating medicine are similar, but u-FSH has economic advantages.

Objective:To investigate the effect of luteinizing hormone (LH) levels on the clinical outcome and cumulative live birth rate of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) fresh embryo transfer in patients with normal ovarian reserve function after application of a flexible protocol of gonadotropin-releasing hormone antagonist (GnRH-A) to promote ovulation with the addition of GnRH-A. Methods:A retrospective cohort study was conducted to analyze the data of 685 patients with normal ovarian reserve function who underwent IVF/ICSI after ovulation induction with antagonist flexible regimen between January 2016 and June 2021 at the Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University. The quartile method was used to group patients after the addition of the antagonist by the mean value of serum LH each time they were monitored (usually twice, depending on the rate of follicular growth and patients' need) until the day of the trigger, and were divided into group A (smaller than the 25th percentile of the mean LH level of the included population, LH<1.25 U/L, n=166), group B (in the 25th to 50th percentile of the mean LH level of the included population, 1.25 U/L≤LH<1.91 U/L, n=174), group C (in the 50th to 75th percentile of the mean LH level of the included population, 1.91 U/L≤LH<2.85 U/L, n=171), and group D (greater than the 75th percentile of the mean LH level of the included population, 2.85 U/L≤LH≤7.55 U/L, n=174). The general condition, clinical data, embryo laboratory indices, clinical outcome indices of fresh embryo transfer cycle and cumulative live birth rate were compared among the 4 groups. Results:After correcting for confounding factors by multifactorial linear regression, the number of high-quality embryos and the rate of blastocyst formation of patients in group C were significantly higher than those in group A, and the differences were statistically significant ( B=0.600, 95% CI: 0.086-1.114, P=0.022; B=0.134, 95% CI: 0.052-0.216, P=0.001). The number of high-quality embryos, the number of available embryos, and the rate of blastocyst formation of patients in group D were significantly higher than those in group A, and the differences were statistically significant ( B=0.771, 95% CI: 0.259-1.284, P=0.003; B=0.730, 95% CI: 0.205-1.255, P=0.007; B=0.085, 95% CI: 0.003-0.167, P=0.042).After multifactorial logistic regression, there was no statistically significant difference in live birth rate between group A and groups B, C and D ( P>0.05). The cumulative live birth rate of patients in group D was significantly higher than that in group A, and the difference was statistically significant ( aOR=2.439, 95% CI: 1.169-4.974, P=0.014). Conclusion:In patients with normal ovarian reserve function, a flexible protocol of antagonists was applied to promote ovulation, and the addition of antagonists had no significant effect on the clinical outcome of the fresh embryo transfer cycle in terms of mean LH levels, but the quality of the embryos was significantly reduced when the mean LH level was <1.25 U/L, and this may consequently reduce the developmental potential of the embryos and the cumulative live birth rate.

Objective:To investigate the effect of luteinizing hormone (LH) levels on the clinical outcome and cumulative live birth rate of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) fresh embryo transfer in patients with normal ovarian reserve function after application of a flexible protocol of gonadotropin-releasing hormone antagonist (GnRH-A) to promote ovulation with the addition of GnRH-A. Methods:A retrospective cohort study was conducted to analyze the data of 685 patients with normal ovarian reserve function who underwent IVF/ICSI after ovulation induction with antagonist flexible regimen between January 2016 and June 2021 at the Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University. The quartile method was used to group patients after the addition of the antagonist by the mean value of serum LH each time they were monitored (usually twice, depending on the rate of follicular growth and patients' need) until the day of the trigger, and were divided into group A (smaller than the 25th percentile of the mean LH level of the included population, LH<1.25 U/L, n=166), group B (in the 25th to 50th percentile of the mean LH level of the included population, 1.25 U/L≤LH<1.91 U/L, n=174), group C (in the 50th to 75th percentile of the mean LH level of the included population, 1.91 U/L≤LH<2.85 U/L, n=171), and group D (greater than the 75th percentile of the mean LH level of the included population, 2.85 U/L≤LH≤7.55 U/L, n=174). The general condition, clinical data, embryo laboratory indices, clinical outcome indices of fresh embryo transfer cycle and cumulative live birth rate were compared among the 4 groups. Results:After correcting for confounding factors by multifactorial linear regression, the number of high-quality embryos and the rate of blastocyst formation of patients in group C were significantly higher than those in group A, and the differences were statistically significant ( B=0.600, 95% CI: 0.086-1.114, P=0.022; B=0.134, 95% CI: 0.052-0.216, P=0.001). The number of high-quality embryos, the number of available embryos, and the rate of blastocyst formation of patients in group D were significantly higher than those in group A, and the differences were statistically significant ( B=0.771, 95% CI: 0.259-1.284, P=0.003; B=0.730, 95% CI: 0.205-1.255, P=0.007; B=0.085, 95% CI: 0.003-0.167, P=0.042).After multifactorial logistic regression, there was no statistically significant difference in live birth rate between group A and groups B, C and D ( P>0.05). The cumulative live birth rate of patients in group D was significantly higher than that in group A, and the difference was statistically significant ( aOR=2.439, 95% CI: 1.169-4.974, P=0.014). Conclusion:In patients with normal ovarian reserve function, a flexible protocol of antagonists was applied to promote ovulation, and the addition of antagonists had no significant effect on the clinical outcome of the fresh embryo transfer cycle in terms of mean LH levels, but the quality of the embryos was significantly reduced when the mean LH level was <1.25 U/L, and this may consequently reduce the developmental potential of the embryos and the cumulative live birth rate.

Objective:To observe the effects of acupuncture and moxibustion on ubiquitin-like containing PHD and RING finger domains 1(UHRF1)and DNA methyltransferase 1(DNMT1)in ectopic endometrium of rats with endometriosis(EMS). Methods:Forty Sprague-Dawley rats were randomly divided into a sham operation group with 10 rats and a model-building group with 30 rats according to body mass.EMS rat models were established in the model-building group and then were divided into a model group,an acupuncture and moxibustion group,and a progesterone group,with 10 rats in each group.All rats were fixed by a fixator.The sham operation group and the model group were given normal saline by gavage.The acupuncture and moxibustion group received acupuncture at Xuehai(SP10)and Sanyinjiao(SP6),moxibustion at Guanyuan(CV4),and gavage of normal saline.The progesterone group was given the mixed liquid made of dydrogesterone and normal saline by gavage.After 28 d of treatments,the three diameters(length,width,and height)of EMS rats'ectopic cysts were measured,the cyst volumes were calculated,the volumes before intervention were subtracted,and the difference values were used to evaluate the growth of ectopic cysts.UHRF1 and DNMT1 mRNA and protein levels in normal endometrium,eutopic endometrium,and ectopic endometrium were detected by real-time polymerase chain reaction and immunohistochemistry. Results:There was no significant difference in the ectopic cyst volume difference between the acupuncture and moxibustion group and the progesterone group(P>0.05),but they were smaller than that of the model group(P<0.05).The levels of UHRF1 and DNMT1 mRNA and protein in the ectopic endometrium of the model group were lower than those in the normal endometrium(P<0.05).The levels of DNMT1 mRNA and UHRF1 protein in the eutopic endometrium of the model group were lower than those in the normal endometrium(P<0.05).The levels of UHRF1 mRNA and protein and the level of DNMT1 protein in the ectopic endometrium of the acupuncture and moxibustion group were higher than those in the model group(P<0.05),and the level of UHRF1 mRNA was higher than that in the progesterone group(P<0.05).The level of DNMT1 mRNA in the eutopic endometrium of the acupuncture and moxibustion group was higher than that in the model group(P<0.05).The levels of UHRF1 and DNMT1 mRNA and protein in the acupuncture and moxibustion group were insignificantly different from those in the normal endometrium(P>0.05). Conclusion:Acupuncture and moxibustion may up-regulate the levels of UHRF1 mRNA and UHRF1 and DNMT1 proteins in the ectopic endometrium to the normal level so as to reduce the volume of ectopic cysts and cure EMS in rats.

Objective:To evaluate the application value of amplicon sequencing in identification of drug-contami-nating microbes in Chinese patent drugs.Methods:Both culturing method and Amplicon Sequencing technology were used to identify bacteria and fungi in 16 batches of Chinese patent drugs.The microbial colonies isolated by culturing medium were identified by 16srDNA/ITS sequencing and MALID-TOF-MS.The amplicon sequencing reads were clustered to OTU(Operational Taxonomic Units)by 97％similarity and BLAST in NCBI to get species taxonomy.Alpha and beta diversity index were used to analyze the microbial community composition in and between batches of drugs.Results:Fifteen species of bacteria and 3 species of fungi were isolated by culturing method,while 75 species of bacteria and 78 species of fungi were identified by amplicon sequencing.Between pre-cultured and non-cultured drug groups,the α diversity index and abundance of most dominant microbial genus were not changed significantly(P＞0.05).Beta diversity analysis showed that microbial diversity between different drugs are more than the same drugs between different pre-treated groups.Conclusion:This study revealed that amplicon sequencing method can provide more comprehensive information on the microbial community composition than tradi-tional culture method.It contributes great value to quick inspection and monitor for microbial contamination in chines patent drugs.

The rapid development of modern industries is accompanied with the aggravating water heavy metal pollution, which poses a potential threat to the aquatic environment and the health of local populations. As an efficient and economical adsorbent, biochar demonstrates the adsorption capacity for heavy metal ions and its adsorption capacity is significantly enhanced after modification. Therefore, biochar can effectively mitigate environmental pollution. By reviewing the existing studies, we summarize the modification methods of biochar, compare the advantages and disadvantages of physical, biological, and chemical modification methods, analyze the effects of modification on the adsorption capacity of biochar for heavy metal ions, and expound the modification mechanism of biochar. On this basis, this article puts forward the future research directions of the application of biochar in treating coexisting pollutants, aiming to provide a reference for the application of biochar in the purification of heavy metal-containing wastewater.
Charcoal/chemistry* ; Metals, Heavy ; Adsorption ; Wastewater/chemistry* ; Water Pollutants, Chemical/chemistry* ; Water Purification/methods* ; Heavy Ions ; Waste Disposal, Fluid/methods*

Objective:To evaluate the application value of amplicon sequencing in identification of drug-contami-nating microbes in Chinese patent drugs.Methods:Both culturing method and Amplicon Sequencing technology were used to identify bacteria and fungi in 16 batches of Chinese patent drugs.The microbial colonies isolated by culturing medium were identified by 16srDNA/ITS sequencing and MALID-TOF-MS.The amplicon sequencing reads were clustered to OTU(Operational Taxonomic Units)by 97％similarity and BLAST in NCBI to get species taxonomy.Alpha and beta diversity index were used to analyze the microbial community composition in and between batches of drugs.Results:Fifteen species of bacteria and 3 species of fungi were isolated by culturing method,while 75 species of bacteria and 78 species of fungi were identified by amplicon sequencing.Between pre-cultured and non-cultured drug groups,the α diversity index and abundance of most dominant microbial genus were not changed significantly(P＞0.05).Beta diversity analysis showed that microbial diversity between different drugs are more than the same drugs between different pre-treated groups.Conclusion:This study revealed that amplicon sequencing method can provide more comprehensive information on the microbial community composition than tradi-tional culture method.It contributes great value to quick inspection and monitor for microbial contamination in chines patent drugs.

Objective:To explore whether the duration of oral estrogen treatment before progesterone application affects neonatal outcome in single frozen-thawed embryo transfer (FET) with artificial cycles.Methods:It was a retrospective cohort study. Patients who underwent in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI), receiving single frozen blastocyst transfer with artificial cycle and delivering a single live birth in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University between January 2015 and December 2019 were included. All FET cycles were divided into four groups according to the estrogen treatment duration before progesterone application, ≤12 d ( n=306), 13-15 d ( n=620), 16-18 d ( n=471), and ≥19 d ( n=275). Primary outcome was the incidence of small for gestational age (SGA). Secondary outcomes were the incidence of preterm birth, low birth weight, macrosomia and large for gestational age (LGA). Results:A total of 1 672 single blastocyst transfer cycles were included. The incidence of SGA among the four groups was 7.8% (24/306), 4.8% (30/620), 5.7% (27/471), and 7.6% (21/275), respectively, with no statistically significant difference ( P=0.204). The results of multiple logistic regression analysis showed that the duration of estrogen used before progesterone application did not affect the incidence of SGA in singleton offspring (with ≤12 d as the reference, 13-15 d: a OR=1.37, 95% CI: 0.70-2.70, P=0.361; 16-18 d: a OR=0.74, 95% CI: 0.40-1.36, P=0.336; ≥19 d: a OR=0.81, 95% CI: 0.44-1.49, P=0.501). There were no significant differences in neonatal preterm birth rate ( P=0.204), low birth weight ( P=0.582), incidences of macrosomia ( P=0.201) and LGA infants ( P=0.335) among the four groups. Conclusion:In artificial FET cycle, the duration of oral estrogen treatment before progesterone application does not affect the outcome of singleton offspring after single blastocyst transfer.

Objective:To explore the clinical application value of frozen-thawed single blastocyst derived from nonpronuclear (0PN) zygotes, monopronuclear (1PN) zygotes and two-pronuclei (2PN) discarded embryos.Methods:A retrospective cohort analysis of the data of patients who underwent frozen-thawed single blastocyst transfer at the Reproduction Center of the Third Affiliated Hospital of Zhengzhou University from March 2014 to November 2020. According to the different sources of transplanted blastocysts, they were divided into 4 groups including group A derived from 2PN available embryos (2PN embryos with morphological rating Ⅰ-Ⅲ), group B derived from 0PN embryos, group C derived from 1PN embryos, group D derived from 2PN discarded embryos (2PN embryos with morphological rating Ⅳ). The basic data of the four groups were analyzed,and group A was used as the reference group to compare the clinical pregnancy outcomes of groups B, C and D, respectively. The neonatal situation was compared at the cycle of singleton live birth. Logistic regression was used to correct for confounding factors and calculate the adjusted odds ratio (a OR) and 95% confidence interval ( CI). Results:After correcting for confounding factors by logistic regression, the live birth rate in group B was significantly lower than that in group A (a OR=0.701, 95% CI: 0.534-0.920, P=0.011). The clinical pregnancy rate and the live birth rate in group D were significantly lower than those in group A (a OR=0.595, 95% CI: 0.456-0.777, P<0.001; a OR=0.600, 95% CI: 0.449-0.800, P=0.001). The differences in miscarriage rate, pregnancy complications and multiple pregnancy rate were not statistically significant compared with group A (all P>0.05). The differences in all the above indicators in group C were not statistically significant compared with group A ( P>0.05). The risk of occurrence of macrosomia (group B: a OR=2.367, 95% CI: 1.299-4.315, P=0.005; group D: a OR=2.711, 95% CI: 1.463-5.026, P=0.002), large for gestational age (group B: a OR=1.930, 95% CI: 1.158-3.217, P=0.012; group D: a OR=2.039, 95% CI: 1.174-3.543, P=0.011) were significantly higher in groups B and D than in group A. The differences in the risk of occurrence of low birth weight, small for gestational age and preterm birth were not statistically significant in groups B and D compared with group A ( P>0.05), and the difference in the risk of occurrence of the above indicators was not statistically significant in group C compared with group A ( P>0.05). Conclusion:When 2PN embryos are not available, abandoned 0PN, 1PN, and 2PN embryos can be transferred after blastocyst culture, but attention should be paid to the increased risk of macrosomia and large for gestational age in offspring.