Objective : To explore the correlation between the clinical, pathological, genetic features, prognosis, and tumor lymph node metastasis in patients with stage ⅠA-Ⅲ B lung invasive non-mucinous adenocarcinoma(INMA). Methods: A retrospective analysis was conducted on 67 eligible patients with INMA. Clinical data, histopathological assessments, and genetic testing were collected. Disease progression-free survival(PFS) was the primary endpoint through follow-up. The chi-square test or Fisher's exact test was used to analyse the correlation between tumour lymph node metastasis and clinicopathological and genetic characteristics. The Cox proportional hazards regression model and Kaplan-Meier method were used to analyse the impact of tumour lymph node metastasis on prognosis. Results: A total of 67 patients were included, aged 46-77 years, with a median age of 61 years. Age, gender, and smoking history were not significantly associated with tumor lymph node metastasis. Larger tumor diameter, tumor progression, and receiving postoperative adjuvant treatment were associated with tumour lymph node metastasis(P<0.05). Poorer differentiated tumors according to International Association for the Study of Lung Cancer(IASLC) grading system was more likely to have lymph node metastasis(P=0.043). There was no significant difference in the types of driver gene mutations and lymph node metastasis. However,EGFRmutations were more common in patients without lymph node metastasis, while co-mutations were more common in patients with lymph node metastasis. Lymph node metastasis was significantly associated with PFS. Patients without lymph node metastasis had a significantly better PFS compared to those with lymph node metastasis(P=0.002). Under different treatment conditions, patients without lymph node metastasis exhibited a significant advantage in PFS when untreated. While treatment showed a trend toward improved PFS, the difference did not reach statistical significance. Additionally, no significant differences in PFS were observed between patients with or without lymph node metastasis following chemotherapy or targeted therapy. Conclusion Lymph node metastasis in INMA patients is related to tumor size, progression status, and gene co-mutations, and is a key prognostic indicator affecting PFS.

Objective: To investigate the correlations among clinicopathological features, histopathological growth patterns and prognosis of extrapulmonary multiple metastatic(stage ⅣB) pulmonary adenocarcinoma with epidermal growth factor receptor(EGFR) mutations. Methods : A total of 488 eligible patients with adenocarcinoma of stage ⅣB. Clinicopathological data,EGFRgene mutation subtypes, metastatic sites, histopathological growth patterns and survival information were collected. The chi-square test(χ2test) and Fisher's exact probability method were used to detect the correlation between the metastasis status and various clinical characteristics; the Kaplan-Meier method was used to conduct survival analysis on the median Progression-Free Survival(PFS) under different clinical characteristics. Cox univariate and multivariate regression analyses were conducted to evaluate the impact of various clinical characteristics on prognosis. Results : The metastatic patterns of stage ⅣB pulmonary adenocarcinoma withEGFRmutations was correlated with histopathological growth patterns(P<0.05). In the group with multiple metastases in a single organ, the proportion of micropapillary type in the group with multiple metastases in a single organ was higher than that in the group with multiple-organ metastases(51.1%vs41.1%), while the proportion of solid type in the group with multiple-organ metastases was higher than that in the group with multiple metastases in a single organ(23.8%vs14.2%). Multiple brain or multiple bone metastases were correlated with histopathological growth patterns and tumor differentiation degree. Compared with the multiple bone metastases group, the proportion of acinar type decreases in the multiple brain metastasis group, while the proportion of micropapillary type increased. Moreover, the proportion of poorly differentiated tumors increased significantly(P<0.05). Compared with multiple bone metastases, the proportion of poorly differentiated tumors significantly increases in the group with multiple brain metastases. The median progression-free survival(PFS) of patients with a predominant solid growth pattern was shorter than that of patients with other growth patterns(12.7 monthsvs17.8 months,P<0.05). The PFS of patients in the poorly differentiated group was worse than that in the moderately differentiated group(15.6 monthsvs17.8 months,P<0.05). There were significant differences in PFS among patients with common sensitive mutations and rare mutationsEGFR(17.3 monthsvs10.2 months,P<0.01). Cox proportional hazards regression model suggested that solid growth pattern, poor differentiation and rare single gene mutation were adverse prognostic factors. Conclusion In stage ⅣB pulmonary adenocarcinoma patients withEGFRmutations, both the metastatic patterns and metastatic sites are significantly correlated with the histopathological growth patterns of tumors. Moreover, theEGFRmutation subtypes as well as the histopathological growth patterns and differentiation degree of tumors significantly affect the prognosis of patients.

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 explore the effect of vaginal microbiota disorder on pregnancy outcomes in the first-time frozen-thawed embryo transfer (FET) patients and perinatal outcomes in single pregnancy live delivery patients.Methods:The clinical data of 2 299 cycles of FET patients in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from January 2021 to December 2022 were collected in a retrospective cohort study. According to the types of dominant bacteria in the vaginal microbiota before transplantation, they were divided into three groups: control group (dominant bacteria were Lactobacillus, which was Gram positive bacteria, 1 849 cycles), the Gram negative bacilli group (191 cycles), and the Gram positive cocci group (259 cycles). Baseline data and pregnancy outcomes were compared among the three groups. The perinatal outcomes of select single pregnancy live birth FET patients who met the inclusion criteria were further analyzed among the three groups. The main observation indicators were live birth rate, miscarriage rate, and preterm birth rate. A multivariate logistic regression model was used to control confounding factors in the main observation indicators, and to analyze the relationship between dominant bacterial types in the vaginal microbiota and live birth rate, miscarriage rate, and preterm birth rate.Results:The difference in endometrial thickness on the day of transplantation among control group, Gram negative bacilli group, and Gram positive cocci group was statistically significant [(9.38±1.58) mm, (9.56±1.70) mm, and (9.84±1.74) mm, respectively, P<0.001]. In the endometrium preparation methods, the proportion of down-regulation+artificial cycle patients in the Gram positive cocci group was higher than that in control group, and the difference was statistically significant [13.13% (34/259) and 7.46% (138/1 849), respectively, P<0.016 7]. In the pregnancy outcomes, there was a statistically significant difference in live birth rate among control group, Gram negative bacilli group, and Gram positive cocci group [49.86% (922/1 849), 49.21% (94/191) and 41.70% (108/259), respectively, P=0.048]. The live birth rate of the Gram positive cocci group was lower than that of control group, and the difference between the two groups was statistically significant ( P<0.016 7). There were no statistically significant differences in other pregnancy outcomes and perinatal outcomes of single pregnancy live birth FET patients (all P>0.05). The multivariate logistic regression model corrected for female age, infertility years, basal follicle stimulating hormone, anti-Müllerian hormone, proportion of single embryo transfer, proportion of single blastocyst transfer, endometrial thickness on transfer day, and endometrial preparation methods, Gram positive cocci were independent risk factors for live brith after FET transplantation (a OR=0.73, 95% CI: 0.55-0.95, P=0.021). Conclusion:The dominant bacteria in the vaginal microbiota before embryo transfer are Gram positive cocci, which may be related to a decrease in live birth rate in first-time FET patients, but not significantly related to the perinatal outcomes.

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 explore the effect of vaginal microbiota disorder on pregnancy outcomes in the first-time frozen-thawed embryo transfer (FET) patients and perinatal outcomes in single pregnancy live delivery patients.Methods:The clinical data of 2 299 cycles of FET patients in the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from January 2021 to December 2022 were collected in a retrospective cohort study. According to the types of dominant bacteria in the vaginal microbiota before transplantation, they were divided into three groups: control group (dominant bacteria were Lactobacillus, which was Gram positive bacteria, 1 849 cycles), the Gram negative bacilli group (191 cycles), and the Gram positive cocci group (259 cycles). Baseline data and pregnancy outcomes were compared among the three groups. The perinatal outcomes of select single pregnancy live birth FET patients who met the inclusion criteria were further analyzed among the three groups. The main observation indicators were live birth rate, miscarriage rate, and preterm birth rate. A multivariate logistic regression model was used to control confounding factors in the main observation indicators, and to analyze the relationship between dominant bacterial types in the vaginal microbiota and live birth rate, miscarriage rate, and preterm birth rate.Results:The difference in endometrial thickness on the day of transplantation among control group, Gram negative bacilli group, and Gram positive cocci group was statistically significant [(9.38±1.58) mm, (9.56±1.70) mm, and (9.84±1.74) mm, respectively, P<0.001]. In the endometrium preparation methods, the proportion of down-regulation+artificial cycle patients in the Gram positive cocci group was higher than that in control group, and the difference was statistically significant [13.13% (34/259) and 7.46% (138/1 849), respectively, P<0.016 7]. In the pregnancy outcomes, there was a statistically significant difference in live birth rate among control group, Gram negative bacilli group, and Gram positive cocci group [49.86% (922/1 849), 49.21% (94/191) and 41.70% (108/259), respectively, P=0.048]. The live birth rate of the Gram positive cocci group was lower than that of control group, and the difference between the two groups was statistically significant ( P<0.016 7). There were no statistically significant differences in other pregnancy outcomes and perinatal outcomes of single pregnancy live birth FET patients (all P>0.05). The multivariate logistic regression model corrected for female age, infertility years, basal follicle stimulating hormone, anti-Müllerian hormone, proportion of single embryo transfer, proportion of single blastocyst transfer, endometrial thickness on transfer day, and endometrial preparation methods, Gram positive cocci were independent risk factors for live brith after FET transplantation (a OR=0.73, 95% CI: 0.55-0.95, P=0.021). Conclusion:The dominant bacteria in the vaginal microbiota before embryo transfer are Gram positive cocci, which may be related to a decrease in live birth rate in first-time FET patients, but not significantly related to the perinatal outcomes.

Objective:To observe the effect of early elevation of luteinizing hormone (LH) with flexible gonadotropin-releasing hormone antagonist (GnRH-A) protocol on pregnancy outcomes of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) fresh embryo transfer. Methods:A retrospective cohort study was conducted to analyze patients with normal ovarian reserve function who underwent IVF/ICSI with flexible GnRH-A protocol at the Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University from January 2016 to September 2021. According to the level of LH before antagonist addition, the patients were divided into two groups, group A: early elevation of LH (LH >10 U/L before the addition of antagonist, n=65), group B: the patients whose LH≤10 U/L before the addition of antagonist ( n=193), which was matched with group A using the 1∶3 propensity score matching by age, duration of infertility, anti-Müllerian hormone, body mass index and antral follicle count. The general conditions, clinical data, embryonic laboratory indicators and clinical outcomes were compared between the two groups. Results:There were no significant differences in the baseline characteristics between the two groups (all P>0.05). There were no significant differences in basic LH levels, gonadotropin (Gn) initiation dosage, total dosage and duration of Gn used, duration of Gn used when adding antagonists and estradiol (E 2) level on the trigger day between group A and group B (all P>0.05). The LH level [12.2 (11.1, 17.5) U/L], E 2 level [3 301.0 (2 708.0, 4 275.0) pmol/L] and follicle diameter [14.0 (12.6, 15.5) mm] were significantly higher in group A than in group B [3.5 (2.2, 5.2) U/L, 2 178.5 (1 208.0, 3 218.0) pmol/L, 13.0 (12.0, 14.0) mm] when adding antagonist (all P<0.001). The level of LH in group A decreased rapidly after adding antagonist, LH level in group A after antagonist application [3.4 (2.0, 5.2) U/L] and the LH level on the trigger day [3.0 (1.7, 4.7) U/L] were still higher than those in group B [2.1 (1.5, 3.3) U/L, P<0.001; 2.1 (1.4, 3.3) U /L, P=0.004], the level of progesterone on the trigger day was not significantly higher than that in group B ( P>0.05). There were no statistically significant differences in the number of oocytes retrieved, two pronuclei fertilization rate, the rate of high-quality embryo, the rate of blastocyst formation, endometrial thickness on the day of transplantation, the number of transferred embryos, the proportion of transferred blastocysts and the implantation rate between the two groups (all P>0.05). The clinical pregnancy rate, the early abortion rate, and the live birth rate were not statistically different between the two groups (all P>0.05). Conclusion:In patients with normal ovarian reserve, LH level was elevated early with antagonist flexible protocol, and decreased rapidly after the timely addition of antagonist, which did not lead to an increase of progesterone on the trigger day, and ultimately did not affect the clinical outcomes of IVF/ICSI fresh embryo transfer.

Objective:To investigate the timing of the first frozen-thawed embryo transfer (FET) on clinical and perinatal outcomes in whole embryo freezing patients, who used the medroxyprogesterone acetate (MPA) in progestin primed ovarian stimulation (PPOS).Methods:A retrospective cohort study was conducted to analyze the clinical data of patients with the first FET after ovulation induction by PPOS protocol in Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University from January 2015 to November 2021. According to the time interval between the day of oocytes retrieved and the day of the first FET in the ovulation induction cycle, they were divided into two groups. Group A: transplantation was performed during the first menstrual cycle after oocyte retrieval (time interval ≥24 d and <45 d, n=80), group B: transplantation was performed at least one month apart after oocytes retrieved (time interval ≥45 d, n=1 040). After propensity score matching (PSM), the general data, clinical excretion promotion, embryo laboratory indicators, indicators related to the first FET cycle, clinical and perinatal outcomes were compared between the two groups. Results:Before PSM, the age of group A was significantly higher than that of group B [38 (35, 41) years vs. 37 (32, 40) years, P=0.020]. After PSM, there were no significant differences in general data, ovulation induction cycle data, embryo laboratory indicators, clinical pregnancy rate, live birth rate and perinatal outcomes between the two groups (all P>0.05). Conclusion:Compared with FET at least one month after oocytes retrieval, FET during the first menstrual cycle after PPOS does not affect clinical or perinatal outcomes. FET can be performed as early as possible after oocytes retrieval according to the condition of patients to shorten the time to reach their first live birth.

Objective:To observe the effect of early elevation of luteinizing hormone (LH) with flexible gonadotropin-releasing hormone antagonist (GnRH-A) protocol on pregnancy outcomes of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) fresh embryo transfer. Methods:A retrospective cohort study was conducted to analyze patients with normal ovarian reserve function who underwent IVF/ICSI with flexible GnRH-A protocol at the Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University from January 2016 to September 2021. According to the level of LH before antagonist addition, the patients were divided into two groups, group A: early elevation of LH (LH >10 U/L before the addition of antagonist, n=65), group B: the patients whose LH≤10 U/L before the addition of antagonist ( n=193), which was matched with group A using the 1∶3 propensity score matching by age, duration of infertility, anti-Müllerian hormone, body mass index and antral follicle count. The general conditions, clinical data, embryonic laboratory indicators and clinical outcomes were compared between the two groups. Results:There were no significant differences in the baseline characteristics between the two groups (all P>0.05). There were no significant differences in basic LH levels, gonadotropin (Gn) initiation dosage, total dosage and duration of Gn used, duration of Gn used when adding antagonists and estradiol (E 2) level on the trigger day between group A and group B (all P>0.05). The LH level [12.2 (11.1, 17.5) U/L], E 2 level [3 301.0 (2 708.0, 4 275.0) pmol/L] and follicle diameter [14.0 (12.6, 15.5) mm] were significantly higher in group A than in group B [3.5 (2.2, 5.2) U/L, 2 178.5 (1 208.0, 3 218.0) pmol/L, 13.0 (12.0, 14.0) mm] when adding antagonist (all P<0.001). The level of LH in group A decreased rapidly after adding antagonist, LH level in group A after antagonist application [3.4 (2.0, 5.2) U/L] and the LH level on the trigger day [3.0 (1.7, 4.7) U/L] were still higher than those in group B [2.1 (1.5, 3.3) U/L, P<0.001; 2.1 (1.4, 3.3) U /L, P=0.004], the level of progesterone on the trigger day was not significantly higher than that in group B ( P>0.05). There were no statistically significant differences in the number of oocytes retrieved, two pronuclei fertilization rate, the rate of high-quality embryo, the rate of blastocyst formation, endometrial thickness on the day of transplantation, the number of transferred embryos, the proportion of transferred blastocysts and the implantation rate between the two groups (all P>0.05). The clinical pregnancy rate, the early abortion rate, and the live birth rate were not statistically different between the two groups (all P>0.05). Conclusion:In patients with normal ovarian reserve, LH level was elevated early with antagonist flexible protocol, and decreased rapidly after the timely addition of antagonist, which did not lead to an increase of progesterone on the trigger day, and ultimately did not affect the clinical outcomes of IVF/ICSI fresh embryo transfer.

Objective:To investigate the timing of the first frozen-thawed embryo transfer (FET) on clinical and perinatal outcomes in whole embryo freezing patients, who used the medroxyprogesterone acetate (MPA) in progestin primed ovarian stimulation (PPOS).Methods:A retrospective cohort study was conducted to analyze the clinical data of patients with the first FET after ovulation induction by PPOS protocol in Reproductive Medicine Center of the Third Affiliated Hospital of Zhengzhou University from January 2015 to November 2021. According to the time interval between the day of oocytes retrieved and the day of the first FET in the ovulation induction cycle, they were divided into two groups. Group A: transplantation was performed during the first menstrual cycle after oocyte retrieval (time interval ≥24 d and <45 d, n=80), group B: transplantation was performed at least one month apart after oocytes retrieved (time interval ≥45 d, n=1 040). After propensity score matching (PSM), the general data, clinical excretion promotion, embryo laboratory indicators, indicators related to the first FET cycle, clinical and perinatal outcomes were compared between the two groups. Results:Before PSM, the age of group A was significantly higher than that of group B [38 (35, 41) years vs. 37 (32, 40) years, P=0.020]. After PSM, there were no significant differences in general data, ovulation induction cycle data, embryo laboratory indicators, clinical pregnancy rate, live birth rate and perinatal outcomes between the two groups (all P>0.05). Conclusion:Compared with FET at least one month after oocytes retrieval, FET during the first menstrual cycle after PPOS does not affect clinical or perinatal outcomes. FET can be performed as early as possible after oocytes retrieval according to the condition of patients to shorten the time to reach their first live birth.