Objective To compare the clinical outcomes of the clomiphene citrate(CC)dual stimulation protocol and the luteal phase ovarian stimulation protocol after natural cycle oocytes retrieved in POSEIDON patients with low expected prognosis.Methods The clinical data from patients who underwent in vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)at the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from May 2016 to December 2023 were analyzed retrospectively.According to the POSEI-DON criteria,including POSEIDON group 3 and group 4,which were treated with the CC dual stimulation protocol(the CC group)and the luteal-phase ovarian stimulation protocol after the natural cycle oocytes retrieval(the natural cycle group).The basic data and clinical outcomes of the two ovulation protocols were analyzed,and the main observations included the number of oocytes retrieved,the cumulative clinical pregnancy rate and the cumulative live birth rate,and explored the impact of ovulation promotion regimens on clinical outcomes through binary logis-tic regression.Results On the whole,there was no significant difference in the total number of oocytes retrieved,cumulative clinical pregnancy rate and live birth rate between the CC group and the natural cycle group(all P>0.05),And in POSEIDON group 4,the numbers of oocytes retrieved in the follicular phase of the CC group were higher than those in the natural cycle group,whereas the number of oocytes retrieved in the luteal phase of natural cycle group were higher,and the difference was statistically significant(P<0.05).After logistic regression analysis,the ovulation stimulation protocols did not affect the cumulative clinical pregnancy rate and live birth rate of POSEI-DON 3 and 4 group(P>0.05).Conclusions For the POSEIDON patients with a low expected prognosis,the clinical effectiveness of the luteal phase ovarian stimulation protocol after natural cycle oocytes retrieved is not inferior to that of the CC dual stimulation protocol.In POSEIDON group 4,on the premise that the AMH levels of patients in the natural cycle group were lower than those in the CC group,who underwent natural cycle ovulation followed by luteal phase induction achieved similar clinical outcomes to those of the CC dual stimulation protocol group.Therefore,the luteal phase ovarian stimulation protocol after natural cycle oocytes retrieved should be more clinically recommended for POSEIDON group 4 patients.

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 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 compare the clinical outcomes of the clomiphene citrate(CC)dual stimulation protocol and the luteal phase ovarian stimulation protocol after natural cycle oocytes retrieved in POSEIDON patients with low expected prognosis.Methods The clinical data from patients who underwent in vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)at the Reproductive Center of the Third Affiliated Hospital of Zhengzhou University from May 2016 to December 2023 were analyzed retrospectively.According to the POSEI-DON criteria,including POSEIDON group 3 and group 4,which were treated with the CC dual stimulation protocol(the CC group)and the luteal-phase ovarian stimulation protocol after the natural cycle oocytes retrieval(the natural cycle group).The basic data and clinical outcomes of the two ovulation protocols were analyzed,and the main observations included the number of oocytes retrieved,the cumulative clinical pregnancy rate and the cumulative live birth rate,and explored the impact of ovulation promotion regimens on clinical outcomes through binary logis-tic regression.Results On the whole,there was no significant difference in the total number of oocytes retrieved,cumulative clinical pregnancy rate and live birth rate between the CC group and the natural cycle group(all P>0.05),And in POSEIDON group 4,the numbers of oocytes retrieved in the follicular phase of the CC group were higher than those in the natural cycle group,whereas the number of oocytes retrieved in the luteal phase of natural cycle group were higher,and the difference was statistically significant(P<0.05).After logistic regression analysis,the ovulation stimulation protocols did not affect the cumulative clinical pregnancy rate and live birth rate of POSEI-DON 3 and 4 group(P>0.05).Conclusions For the POSEIDON patients with a low expected prognosis,the clinical effectiveness of the luteal phase ovarian stimulation protocol after natural cycle oocytes retrieved is not inferior to that of the CC dual stimulation protocol.In POSEIDON group 4,on the premise that the AMH levels of patients in the natural cycle group were lower than those in the CC group,who underwent natural cycle ovulation followed by luteal phase induction achieved similar clinical outcomes to those of the CC dual stimulation protocol group.Therefore,the luteal phase ovarian stimulation protocol after natural cycle oocytes retrieved should be more clinically recommended for POSEIDON group 4 patients.

Rejection and adverse reactions caused by long-term use of immunosuppressants severely affect the survival rate and quality of life of organ transplant recipients. Immune tolerance induction plays a key role in improving the survival rate and quality of life of organ transplant recipients. In recent years, tremendous progress has been achieved in adoptive re-transfusion of regulatory cells. In this article, research progress in regulatory T cell (Treg), myeloid-derived suppressor cell (MDSC) and regulatory B cell (Breg) in animal experiment and clinical application was reviewed, and the main clinical problems of adoptive re-transfusion of regulatory cells, the application of chimeric antigen receptor Treg and the concept of cell therapy in immune evaluation were summarized, aiming to deepen the understanding of regulatory cell therapy, promote the application of regulatory cells in immune tolerance of organ transplantation, and improve clinical efficacy of organ transplantation and the quality of life of recipients.

Objective To investigate the role of tolerogenic dendritic cell (tolDC) in inducing immune tolerance in liver transplantation. Methods Liver transplantation rat models of spontaneous tolerance [Brown Norway (BN)→Lewis, tolerance group, n=6] and acute rejection (AR) (Lewis→BN) were established. In AR rat models, tolDC transfusion was performed in the study group (tolDC group, n=6) and no intervention was given in the control group (AR group, n=6). The survival time of rats in each group was observed. The transplant liver tissues of rats were prepared for pathological examination in each group. The expression of myeloid dendritic cell (mDC) and plasmacytoid dendritic cell (pDC) in rat peripheral blood, transplant liver, spleen and lymph nodes in each group was detected by flow cytometry. The expression levels of serum interleukin (IL)-10 and interferon (IFN)-γ in each group were measured by enzyme-linked immune absorbent assay. Results Pathological manifestations of rats in the AR group mainly included inflammatory cell infiltration and tissue structural disorder in transplant liver, and the survival time was 7-14 d. In the tolDC and tolerance groups, the transplant liver tissues were almost normal, and the longest survival time exceeded 100 d. Compared with the AR group, the expression levels of CD11⁺mDC in peripheral blood, transplant liver, spleen and lymph nodes of rats were significantly down-regulated in the tolerance and tolDC groups (all P < 0.05), and those of CD86 and major histocompatibility complex (MHC)Ⅱon the surface of CD11⁺mDC were also significantly down-regulated (all P < 0.05). Compared with the AR group, the expression levels of pDC in peripheral blood, transplant liver, spleen and lymph nodes of rats were significantly up-regulated in the tolerance and tolDC groups (all P < 0.05), whereas those of MHCⅡon the surface of pDC were all significantly down-regulated (all P < 0.05). Compared with the AR group, the expression levels of serum IL-10 were significantly up-regulated, and IFN-γ were significantly down-regulated in the tolerance and tolDC groups (all P < 0.05). Conclusions As tolDC subsets, mDC and pDC play a positive role in regulating the incidence of graft immune tolerance in rats after liver transplantation.

Objective To investigate the guiding role of quantitative evaluation system of immune status in the individualized management of immunosuppressants for the recipients after liver transplantation. Methods Clinical data of 239 liver transplant recipients were retrospectively analyzed. MingDao Immune Cell Analysis (MICA) was established. All recipients were divided into two groups according to the adjustment regimens of immunosuppressants. The immunosuppressant regimen was adjusted according to MingDao Immune System Score (MISS) in the MISS group (n=84), and the medication plan was empirically adjusted during the same period in the control group (n=155). According to the time of postoperative detection (t), the recipients in the MISS group were divided into subgroup A (t ≤ 28 d, n=78), subgroup B (28 d < t ≤ 6 months, n=68), subgroup C (6 months < t ≤ 12 months, n=18), subgroup D (12 months < t ≤ 24 months, n=18) and subgroup E (t > 24 months, n=19). In the MISS group, postoperative MISS scores of recipients in subgroups A-E were analyzed. The incidence of acute rejection and opportunistic infection and the overall survival rate were statistically compared between the MISS and control groups. Results The MISS scores in subgroups A-E were -7.0 (-13.2, -2.0), -2.0 (-5.8, 1.8), -0.5 (-7.3, 2.8), -2.0 (-4.5, 3.3) and -3.0 (-6.0, 1.0), respectively. The immune status of the recipients was gradually improved over postoperative time, and the difference between two groups was statistically significant (P < 0.05). In the MISS group, 15% (13/84) of the recipients developed acute rejection, and 27% (42/155) in the control group, and the difference was statistically significant (P < 0.05). In the MISS group, the MISS score of the recipients with acute rejection was 0 (-2.5, 3.5), and -5.0 (-12.0, -1.0) for their counterparts without acute rejection, and the difference was statistically significant (P < 0.05). In the MISS group, 2% (2/84) of the recipients presented with postoperative opportunistic infection, and 9% (14/155) in the control group, and the difference was statistically significant (P < 0.05). In the MISS group, the 1- and 3-year overall survival rates were 86.9% and 79.8%, and 83.2% and 76.8% in the control group, and no significant difference was observed between two groups (P > 0.05). Conclusions MICA and MISS score may reflect the immune status of liver transplant recipients, and guide the individualized management of administration of immunosuppressants after liver transplantation.

Objective To evaluate the changes and significance of lymphocyte subsets in the recipients with acute rejection after liver transplantation. Methods The recipients presenting with acute rejection after liver transplantation were assigned into the rejection group (n=17), and their counterparts with stable liver function were allocated into the control group (n=17) according to the ratio of 1∶1 by propensity score matching method. The incidence of acute rejection after liver transplantation was analyzed, and the concentration of tacrolimus in the recipients was compared between two groups. The absolute value and proportion of lymphocyte subsets in peripheral blood were compared between two groups. The diagnostic value of lymphocyte subsets for acute rejection after liver transplantation was assessed by the receiver operating characteristic (ROC) curve. The absolute value and proportion of lymphocyte subsets in the rejection group were compared before and after treatment. Results Among 17 recipients in the rejection group, 4 cases developed acute rejection within postoperative 28 d, and 13 cases had acute rejection within postoperative 29-180 d. No significant difference was noted in the tacrolimus concentration between two groups (P=0.295). Compared with the control group, the proportions of peripheral blood T cells, CD4⁺T cells, B cells and natural killer (NK) T cells were significantly increased in the rejection group (all P < 0.05). The elevated proportion of NKT cells in the early stage after liver transplantation was an independent risk factor for acute rejection following liver transplantation[odds ratio (OR) 1.774, 95% confidence interval (CI) 1.059-2.971, P=0.029]. ROC curve analysis showed that the area under curve (AUC) of CD4⁺T cells, B cells and NKT cells was 0.76, 0.73 and 0.77, respectively. The AUC of combined use of CD4⁺T cells, B cells and NKT cells was 0.89, with a cut-off value of 0.69, sensitivity of 0.706 and specificity of 0.941. After corresponding treatment, all recipients were gradually recovered, and liver functions were eventually restored to normal in the rejection group. After treatment, the proportion of T cells, CD4⁺T cells, CD8⁺T cells and NK cells was significantly decreased (all P < 0.05). Conclusions The elevated proportion of NKT cells indicates an increased risk of acute rejection after liver transplantation. Combined use of CD4⁺T cells, B cells and NKT cells may deliver early detection and diagnosis of acute rejection after liver transplantation.

To study the epidemiology and etiology characteristics of first imported Chikungunya fever case in Henan province, China, 2017. The patient was confirmed by Chikungunya virus (CHIKV) infected as CHIKV ribonucleotide was continuously detected in his serum specimens. BHK-21 cell line was used for virus isolation, the strain was named CHIKV/Henan001/2017. CHIKV/Henan001/2017 belonged to genotype ECSA. The highest ribonucleotide homology sequence of highly conserved region E1 with CHIKV/Henan001/2017 was hk02 strain (99.8%), who was an imported strain to Hong Kong, China, 2016. Epidemiological information and laboratory testing confirmed it was an imported Chikungunya fever case in Henan province, 2017. No secondary case has been reported.

Objective: To confirm the laboratory diagnosis of dengue bordline cases reported in Henan Province and trace its origin from molecular level in 2017. Methods: The study samples were blood samples (3-5 ml), which came from 8 suspected cases of dengue fever reported in the 2017 direct reporting system of Henan provincial infectious disease monitoring network. Meanwhile, case investigation was conducted according to National dengue fever surveillance programme. Serum were separated from blood samples and tested for Dengue NS1 antigen, IgM & IgG antibodies, and dengue RNA. According to dengue diagnosis criteria, confirmed cases were identified by testing results. Samples carried dengue RNA performed for real-time PCR genotyping and amplification of E gene. Then, the amplicons were sequenced and homological and phylogenetic analyses were constructed. Results: 8 serum samples of suspected dengue cases were collected in Henan Province, 2017. Six of them were diagnosed as dengue confirmed cases. All the dengue confirmed cases belonged to outside imported cases, 5 of them were positive by dengue RNA testing. Genotyping results showed there were 1 DENV1 case, 2 DENV2 cases and 2 DENV3 cases. A DENV2 case and a DENV3 case of this study were traced its origin successfully. The sequence of Pakistan imported DENV2 case belongs to cosmopolitan genotype, which was the most consistent with Pakistan's DENV2 KJ010186 in 2013 (identity 99.0%). The sequence of Malaysia imported DENV3 case belongs to genotype I, which was the most consistent with Singapore's DENV3 KX224276 in 2014(identity 99.0%). Conclusion The laboratory diagnosis and molecular traceability of dengue cases in Henan Province in 2017 confirmed that all cases were imported and did not cause local epidemics.