The role of age and AMH on cumulative live birth rates over multiple frozen-thawed embryo transfer cycles: a study based on low prognosis patients of POSEIDON 3 and 4 groups.

BACKGROUND: Among the POSEIDON criteria, group 3 and group 4 have an expected low prognosis. For those patients with inadequate ovary reserve, embryo accumulated from consecutive oocyte retrieval cycles for multiple frozen-thawed embryo transfers (FET) has become more common. It is necessary to inform them of the pregnancy outcomes after single or multiple FET cycles before the treatment. However few studies about cumulative live birth rate (CLBR) for those with low prognosis have been reported. METHODS: This retrospective study included 4712 patients undergoing frozen embryo transfer cycles from July 2015 to August 2020. Patients were stratified as POSEIDON group 3, group 4, control 1 group (< 35 years) and control 2 group (≥ 35 years). The primary outcome is CLBRs up to six FET cycles and the secondary outcomes were LBRs per transfer cycle. Optimistic approach was used for the analysis of CLBRs and the depiction of cumulative incidence curves. RESULTS: Under optimistic model analyses, control 1 group exhibited the highest CLBR (93.98%, 95%CI 91.63-95.67%) within 6 FET cycles, followed by the CLBR from women in POSEIDON group 3(92.51%, 95%CI 77.1-97.55)was slightly lower than that in control 1 group. The CLBR of POSEIDON group 4(55% ,95%CI 39.34-70.66%)was the lowest and significantly lower than that of control 2 group(88.7%, 95%CI 80.68-96.72%). Further, patients in POSEIDON group 4 reached a CLBR plateau after 5 FET cycles. CONCLUSIONS: The patients of POSEIDON group 3 may not be considered as traditional "low prognosis" in clinical practice as extending the number of FET cycles up to 6 can archive considerably CLBR as control women. While for the POSEIDON group 4, a simple repeat of the FET cycle is not recommended after four failed FET cycles, some strategies such as PGT-A may be beneficial.

A novel technique for avoidance of sternotomy, diaphragmic incision and cardiopulmonary bypass during cavoatrial tumor thrombectomy for renal cell carcinoma with intraatrial tumor thrombus: a case series at a single center.

BACKGROUND: Radical nephrectomy with thrombectomy in patients with renal cell carcinoma (RCC) and level IV thrombus extending to the right atrium (RA) offers improved survival. However, this procedure is associated with significant perioperative morbidity and mortality. In this report, we describe a novel milking technique for patients with RA tumor thrombus using abdominal access, which does not require diaphragmic incision, sternotomy, right atriotomy, or cardiopulmonary bypass (CPB). METHODS: Between January 2019 and January 2022, four patients underwent resection of renal cell carcinoma extending into RA by a milking technique developed to avoid diaphragmic incision, sternotomy, or CPB. Patient characteristics, perioperative data, pathological features, and survival were evaluated. RESULTS: Complete resection was successful through pure transabdominal access without diaphragmic incision, sternotomy, or CPB in all patients. CONCLUSION: We conclude that radical nephrectomy and thrombectomy in optimized cases with renal cell carcinoma extending into RA can be safely and effectively performed without diaphragmic incision, sternotomy, or CPB, avoiding serious perioperative complications while providing acceptable oncological outcomes.

Influence of ambient air pollution on successful pregnancy with frozen embryo transfer: A machine learning prediction model.

Numerous air pollutants have been reported to influence the outcomes of in vitro fertilization (IVF). However, whether air pollution affects implantation in frozen embryo transfer (FET) process is under debate. We aimed to find the association between ambient air pollution and implantation potential of FET and test the value of adding air pollution data to a random forest model (RFM) predicting intrauterine pregnancy. Using a retrospective study of a 4-year single-center design，we analyzed 3698 cycles of women living in Shanghai who underwent FET between 2015 and 2018. To estimate patients' individual exposure to air pollution, we computed averages of daily concentrations of six air pollutants including PM2.5, PM10, SO2, CO, NO2, and O3 measured at 9 monitoring stations in Shanghai for the exposure period (one month before FET). Moreover, A predictive model of 15 variables was established using RFM. Air pollutants levels of patients with or without intrauterine pregnancy were compared. Our results indicated that for exposure periods before FET, NO2 were negatively associated with intrauterine pregnancy (OR: 0.906, CI: 0.816-0.989). AUROC increased from 0.712 to 0.771 as air pollutants features were added. Overall, our findings demonstrate that exposure to NO2 before transfer has an adverse effect on clinical pregnancy. The performance to predict intrauterine pregnancy will improve with the use of air pollution data in RFM.

Does surgery benefit patients with oligometastatic or metastatic prostate cancer? - A retrospective cohort study and meta-analysis.

BACKGROUND: To evaluate long-term oncological outcomes of radical prostatectomy (RP) plus androgen deprivation therapy (ADT) in oligometastatic prostate cancer (PCa) patients. METHODS: Our study included oligometastatic PCa patients hospitalized between January 1, 2010 and December 31, 2015, who received ADT with or without RP. We evaluated survival by employing Kaplan-Meier methods, with log-rank tests and univariate and multivariate Cox regression analyses. A meta-analysis of previously published studies was additionally performed. RESULTS: The median follow-up times of both groups were 68.4 months (interquartile range = 56.5-85.0). In this cohort study, significant statistical difference in preoperative total prostate-specific antigen (tPSA; p = .121), clinical T stage (p = .115), and N stage (p = .394) were not found between the two groups. Meanwhile, the difference in overall survival (OS) between the two groups did not reach statistical significance (p = .649). A significant difference was not observed in castration-resistant prostate cancer (CRPC)-free survival between two groups as well (p = .183). Numbers of metastases might be an independent prognosis factor (p = .05) for OS, and postoperative tPSA is a risk predictor for CRPC-free survival (p = .032). A meta-analysis of four relevant studies demonstrated significant statistical difference in clinical improvement with RP plus ADT over ADT alone in OS survival (p < .001; hazard ratio [HR] = 0.51; 95% confidence interval [CI] = 0.38-0.69) instead of CRPC-free survival (p = .42; HR = 0.86; 95% CI = 0.59-1.24). CONCLUSION: The addition of RP to ADT for the treatment of oligometastatic PCa was associated with an improved OS instead of CRPC-free survival.

TMT-based proteomic and bioinformatic analyses of human granulosa cells from obese and normal-weight female subjects.

BACKGROUND: Increasing evidence supports a relationship between obesity and either infertility or subfertility in women. Most previous omics studies were focused on determining if the serum and follicular fluid expression profiles of subjects afflicted with both obesity-related infertility and polycystic ovary syndrome (PCOS) are different than those in normal healthy controls. As granulosa cells (GCs) are essential for oocyte development and fertility, we determined here if the protein expression profiles in the GCs from obese subjects are different than those in their normal-weight counterpart. METHODS: GC samples were collected from obese female subjects (n = 14) and normal-weight female subjects (n = 12) who were infertile and underwent in vitro fertilization (IVF) treatment due to tubal pathology. A quantitative approach including tandem mass tag labeling and liquid chromatography tandem mass spectrometry (TMT) was employed to identify differentially expressed proteins. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were then conducted to interrogate the functions and pathways of identified proteins. Clinical, hormonal, and biochemical parameters were also analyzed in both groups. RESULTS: A total of 228 differentially expressed proteins were noted, including 138 that were upregulated whereas 90 others were downregulated. Significant pathways and GO terms associated with protein expression changes were also identified, especially within the mitochondrial electron transport chain. The levels of free fatty acids in both the serum and follicular fluid of obese subjects were significantly higher than those in matched normal-weight subjects. CONCLUSIONS: In GCs obtained from obese subjects, their mitochondria were damaged and the endoplasmic reticulum stress response was accompanied by dysregulated hormonal synthesis whereas none of these changes occurred in normal-weight subjects. These alterations may be related to the high FFA and TG levels detected in human follicular fluid.

TRIB3 regulates FSHR expression in human granulosa cells under high levels of free fatty acids.

BACKGROUND: Granulosa cells (GCs) in cumulus oophorus highly express follicle stimulating hormone receptor (FSHR), which is the most important mediator of both estradiol synthesis and oocyte maturation. Obese women have elevated free fatty acids (FFAs) levels in their follicular fluids and decreased FSHR expression in GCs, which is related to an altered protein kinase B/glycogen synthase kinase 3ß (Akt/GSK3ß) signaling pathway. Such FFA increases accompany 3-fold rises in pseudokinase 3 (TRIB3) expression and reduce the Akt phosphorylation status in both the human liver and in insulinoma cell lines. Therefore, in a high FFA environment, we determined if TRIB3 mediates regulation of FSHR via the Akt/GSK3ß signaling pathway in human GCs. METHODS: GCs from women undergoing in vitro fertilization were collected and designated as high and low FFAs cohorts based on their follicular fluid FFA content. GCs with low FFA levels and a human granulosa-like tumor (KGN) cell line were exposed to palmitic acid (PA), which is a dominate FFA follicular fluid constituent. The effects were assessed of this substitution on the Akt/GSK3ß signaling pathway activity as well as the expressions of TRIB3 and FSHR at both the gene and protein levels by qPCR, Western blot and immunofluorescence staining analyses. Meanwhile, the individual effects of TRIB3 knockdown in KGN cells and p-AKT inhibitors were compared to determine the mechanisms of FFA-induced FSHR downregulation. RESULTS: The average FSH dose consuming per oocyte (FSH dose/oocyte) was elevated and Top embryo quality ratio was decreased in women with high levels of FFAs in their follicular fluid. In these women, the GC TRIB3 and ATF4 protein expression levels were upregulated which was accompanied by FSHR downregulation. Such upregulation was confirmed based on corresponding increases in their gene expression levels. On the other hand, the levels of p-Akt decreased while p-GSK3ß increased in the GCs. Moreover, TRIB3 knockdown reversed declines in FSHR expression and estradiol (E2) production in KGN cells treated with PA, which also resulted in increased p-Akt levels and declines in the p-GSK3ß level. In contrast, treatment of TRIB3-knockdown cells with an inhibitor of p-Akt (Ser473) resulted in rises in the levels of both p-GSK3ß as well as FSHR expression whereas E2 synthesis fell. CONCLUSIONS: During exposure to a high FFA content, TRIB3 can reduce FSHR expression through stimulation of the Akt/GSK3ß pathway in human GCs. This response may contribute to inducing oocyte maturation.

Adjuvant instant preoperative renal artery embolization facilitates the radical nephrectomy and thrombectomy in locally advanced renal cancer with venous thrombus: a retrospective study of 54 cases.

BACKGROUND: The role of renal artery embolization (RAE) in the therapeutic armamentarium is always controversial. The present study aimed to assess the safety and the surgical outcomes of the instant renal artery embolization (I-RAE) prior to nephrectomy and thrombectomy in patients with locally advanced renal cell carcinoma (RCC) with venous thrombus. METHODS: We performed a retrospective analysis of 54 patients treated with nephrectomy and thrombectomy between January 2012 and January 2019. Twenty-four patients were treated with I-RAE before surgery. Thirty patients received surgery alone (non-RAE group). The patient demographics, operation time, blood loss, transfusion requirements, complications, and other surgical parameters were analyzed between the two groups. RESULTS: The mean tumor size in the I-RAE group was significantly larger than that in the non-RAE group (11.1 cm versus 7.9 cm; p = .001). The mean estimated blood loss was significantly lower in the I-RAE group compared to that in the non-RAE group (596 ml versus 827 ml; p = .015), and the patients in the non-RAE group were more likely to receive blood transfusion (red blood cell, RBC units, 4 U versus 6 U, p = .025; plasma volume, 200 ml versus 400 ml, p = .01). No differences were found in operative duration, ICU stay, perioperative complications, and length of postoperative hospitalization. CONCLUSIONS: Instant preoperative adjuvant renal artery embolization (I-RAE) is a safe technique. It facilitates nephrectomy and thrombectomy by reducing blood loss, transfusion requirements, and complications of delayed operations, providing urologists with a reliable option for treatment of locally advanced RCC with tumor thrombus.

Emergency retroperitoneal laparoscopic partial nephrectomy for ruptured renal angiomyolipomas: a retrospective single-center series of 15 cases.

BACKGROUND: To assess the safety, tumor control and renal function preservation of the emergency retroperitoneal laparoscopic partial nephrectomy (LPN) for ruptured renal angiomyolipoma (AML) and summarize our single-center initial experience. METHODS: We performed a retrospective analysis of 15 patients pathologically confirmed renal AML treated with emergency retroperitoneal LPN between January 2016 and May 2019. The patient demographics, operation time, blood loss, transfusion requirements, complications and other surgical parameters were analyzed. Follow-up was performed by serum creatinine and imaging modalities. RESULTS: Fifteen patients were performed with emergency LPN with the median age 41.6 years. The mean size of the renal AMLs was 7.8 cm. The mean size of the retroperitoneal hematomas was 8.5 cm. All the emergency surgeries were performed successfully without any conversion to nephrectomy or open surgery. The mean operative time was 101 min. The mean warm ischemia time was 28 min. The mean estimated blood loss was 311 ml. Five patients required intraoperative blood transfusion (33.3%, 5/15). The mean transfused RBC was 4 U (range 2-6 U), and the mean transfused plasma was 200 ml (range 200-400 ml). The mean drainage duration was 3 days (range 2-5 days). The mean postoperative hospitalization was 4.7 days. No patients experienced intraoperative complications. The mean serum creatine was slightly higher after surgery (53.1 vs. 55.9 µmol/L). One patient had postoperative perirenal fluid collection. No patients needed dialysis. No recurrence was observed in the patients at the median follow-up of 24.1 months. CONCLUSIONS: Our initial experience shows that the emergency retroperitoneal LPN is a safe, minimally invasive procedure for emergency patients with ruptured renal AMLs. It could be considered as an effective alternative to renal artery embolization in selected emergency patients.

Endometrium cytokine profiles are altered following ovarian stimulation but almost not in subsequent hormone replacement cycles.

PURPOSE: To investigate the impact of ovarian stimulation and hormone replacement treatment on key regulatory cytokines in endometrial secretion during endometrium implantation window. METHODS: Fifty-six patients undergoing ovarian stimulation (OS) with gonadotropin releasing hormone antagonist and frozen embryo transfer with hormone replacement treatment (HRT) were recruited. Endometrial secretion aspiration was performed repeatedly during implantation window in natural, OS and HRT cycles of every patient. The concentrations of 17 mediators, known to be involved in human embryo implantation, were assessed by multiplex immunoassay. RESULTS: Compared with natural cycle (NC), the concentration of IFN-γ, G-CSF and IL-8 within endometrium were almost the same following OS and HRT. Furthermore, increased MCP-1 levels were observed following HRT and OS. In addition, an increase in IL-1b, IL-7, IL-17, IL-6, TNF-a, IL-12, IL-4, IL-13, IL-10, IL-5, VEGF and MIP-1b concentrations were found in OS cycle only. The level of GM-CSF was lower in HRT cycle and higher in OS cycle, when compared with NC. Among all 17 cytokines, no correlation was found between cytokine concentrations and serum estradiol and progesterone, while only IL-7 concentration has a low correlation with serum LH level. CONCLUSION: Compared to natural and hormone replacement cycle, patients' endometrium cytokine profiles present an increased inflammatory response following ovarian stimulation.

Comparison of clinical outcomes among dual ovarian stimulation, mild stimulation and luteal phase stimulation protocols in women with poor ovarian response.

This study aimed to determine whether consecutive ovarian stimulation in follicular and luteal phases within a single menstrual cycle (dual stimulation) is achievable and superior to conventional stimulation for poor ovarian responders (PORs). Data of 260 PORs were retrospectively collected and divided into three groups. Group A comprised of cycles with dual ovarian stimulation (n = 76), which were divided into two subgroups (follicular [group A-F] and luteal phase stimulation [group A-L]); group B comprised of cycles with ovarian stimulation that was performed only in the luteal phase (n = 52). Group C comprised of mild ovarian stimulation cycles (n = 132). Baseline parameters were not different among the three groups. The numbers of oocytes and embryo obtained were less in group A-F than group B and C, while group A overall had significantly more oocytes and viable embryo retrieved than did group B and C. Group A-L consumed significantly less gonadotropin than group B, without compromising the number of retrieved oocytes and embryo. The pregnancy outcomes of transfer of embryo from different stimulation phases were similar. We conclude that dual ovarian stimulation protocol is effective and potentially optimal for PORs.

Follicular fluid cytokine composition and oocyte quality of polycystic ovary syndrome patients with metabolic syndrome undergoing in vitro fertilization.

PURPOSE: To provide insights into the correlation among lipid metabolism, cytokine profiles in the follicular fluid (FF) and embryo quality of women with Polycystic Ovary Syndrome (PCOS) with metabolic syndrome (MS). METHODS: Ninety women undergoing in vitro fertilization (IVF) treatment were recruited, including 60 PCOS patients (PCOS non MS and PCOS MS) and 30 age-matched controls. Individual FF samples were analyzed using the cytometric multiplex immunoassay. RESULTS: In the FF, the PCOS MS group was associated with higher, total cholesterol (TC), triglyceride (TG) and lower high-density lipoprotein (HDL) concentrations compared with that in the control group (P<0.05). The FF tumor necrosis factor-α (TNF-α) level in the PCOS MS group was 3.89±1.18ng/mL, which was significantly higher compared with the control (2.94±1.02ng/mL) and PCOS non-MS groups (3.05±1.21ng/mL) (P=0.002), while the granulocyte colony-stimulating factor (G-CSF) level in the PCOS MS group (4.18±1.33ng/mL) was lower compared with the control (5.61±1.82ng/mL) and PCOS non-MS groups (5.32±1.91ng/mL) (P=0.004). The FF G-CSF showed a trend toward negative relationship with TG and TC; TNF-α concentration was positively associated with TG. The percentage of top-quality embryo decreased in the PCOS MS group than in the other two groups (20% vs. 38.4% and 34.6%). CONCLUSIONS: In conclusion, there was an elevated lipolysis condition within the FF of PCOS MS patients and the TNF-α and G-CSF levels in FF were associated with top-quality embryo percentage. TNF-α and G-CSF may be the key cytokines involved in the mechanism of decreased embryo development potential in PCOS MS patients.

CYLD downregulates Livin and synergistically improves gemcitabine chemosensitivity and decreases migratory/invasive potential in bladder cancer: the effect is autophagy-associated.

Although GC (gemcitabine and cisplatin) chemotherapy remains an effective method for treating bladder cancer (BCa), chemoresistance is a major obstacle in chemotherapy. In this study, we determined whether gemcitabine resistance correlates with migratory/invasive potential in BCa and whether this relationship is regulated by the cylindromatosis (CYLD)-Livin module. First, we independently investigated the correlation of CYLD/Livin and gemcitabine resistance with the potential for tumor migration and invasiveness. Second, we found that co-transfected CYLD and Livin dramatically improved sensitivity to gemcitabine chemotherapy and decreased migration/invasion potential. Next, we determined that CYLD may regulate Livin by the NF-κB-dependent pathway. We also found that CYLD overexpression and Livin knockdown might improve gemcitabine chemosensitivity by decreasing autophagy and increasing apoptosis in BCa cells. Finally, the effects of CYLD-Livin on tumor growth in vivo were evaluated. Our study demonstrates that CYLD-Livin might represent a potential therapeutic for chemoresistant BCa.

Treatment of Renal Stones ≥20 mm with Extracorporeal Shock Wave Lithotripsy.

AIMS: To identify subgroups of patients with renal stones ≥20 mm that are more suitable for extracorporeal shock wave lithotripsy (ESWL) monotherapy. METHODS: A total of 376 patients with renal stones ≥20 mm underwent monotherapy with ESWL. The treatment outcome was evaluated after 3 months of follow-up. A stone-free status or fragmentation of stones to 4 mm or smaller was considered efficacious. RESULTS: At 3 months after treatment, the overall stone-free rate was 64.4%, and the efficacy rate was 70.7%. The efficacy rate was 89.4% for patients with a residual stone surface area ≤50% of baseline after the first ESWL, while the efficacy rate was 32.4% for other patients. The efficacy was 92.2% for stones ≤400 mm2 and those with lower radiodensity, as determined by a plain (KUB) film. CONCLUSIONS: For renal stones with a surface area ≤400 mm2 and a radiodensity equal to or less than that of the 12th rib as determined by a KUB film, ESWL may be considered the first line of treatment, even for stones with a diameter ≥20 mm. For large stones requiring repeat treatments, the surface area of the residual stones after the first ESWL is a predictor of the final treatment result.

Elevated histone deacetylase 10 expression promotes the progression of clear cell renal cell carcinoma by Notch-1-PTEN signaling axis.

BACKGROUND: Clear cell renal cell carcinoma (ccRCC), the most common pathological subtype of kidney cancer, accounts for approximately 70% to 80% of all cases. Histone deacetylase 10 (HDAC10) belongs to the HDAC class IIb subgroup, one of the histone deacetylases (HDAC) family. Previous studies suggest that HDAC10 may regulate the development of multiple tumor types. The specific molecular mechanisms employed by HDAC10 in the etiology of ccRCC still need to be discovered. METHODS: The analysis included examining HDAC10 expression levels and their clinical importance within a cohort of inpatients and ccRCC patients documented in the Tumor Genome Atlas (TCGA). Moreover, the biological functions and underlying molecular mechanisms of HDAC10 were investigated. RESULTS: HDAC10 showed increased expression in ccRCC tumor tissues. Subsequent analysis revealed overexpression of HDAC10 was associated with advanced clinical phenotype and unfavorable prognosis. The absence of HDAC10 significantly decreased ccRCC cell proliferation and migration capabilities. Mechanistic research suggests that HDAC10 may promote RCC development by activating the Notch-1 pathway and downregulating PTEN expression levels. CONCLUSION: In summary, HDAC10 can modulate critical biological processes in ccRCC, including proliferation, migration, and apoptosis. Notably, the Notch-1 pathway and PTEN serve as crucial signaling pathways and target genes through which HDAC10 regulates the progression of ccRCC. These findings offer a novel outlook for ccRCC treatment.

Long-term pituitary downregulation before frozen embryo transfer could improve pregnancy outcomes in women with adenomyosis.

Some studies have shown that long-term gonadotropin-releasing hormone (GnRH) agonist administration before in vitro fertilization/intracytoplasmic sperm in infertile women with endometriosis or adenomyosis significantly increases the chances of pregnancy. We were interested in whether long-term GnRH agonist pretreatment could improve pregnancy outcomes in adenomyosis patients undergoing frozen embryo transfer (FET) after preparation of the endometrium with hormone replacement therapy (HRT). Totally, 339 patients with adenomyosis were included in this retrospective study, 194 received long-term GnRH agonist plus HRT (down-regulation + HRT) and 145 received HRT. There were no differences between the groups in characteristic such as age, body mass index, duration or cause of infertility, serum CA-125 level and basal hormone levels. On the day of progesterone administration, mean endometrial thickness and serum progesterone level were significantly greater in HRT patients. Mean score and number of embryos transferred showed no differences. In down regulation + HRT group, clinical pregnancy, implantation and ongoing pregnancy rates were 51.35%, 32.56% and 48.91%, respectively, significantly higher than that of HRT group (24.83%, 16.07% and 21.38%, respectively). So, we concluded that in FET, long-term GnRH agonist pretreatment significantly improved pregnancy outcomes in patients with adenomyosis.

Type 2 diabetes mellitus and the risk of male infertility: a Mendelian randomization study.

Objective: To assess the causal effect of type 2 diabetes mellitus (T2DM) on male infertility (MI) and erectile dysfunction (ED) by Mendelian randomization (MR) analysis. Methods: Data for T2DM, MI, and ED were obtained from genome-wide association studies (GWAS) involving 298, 957, 73, 479, and 223, 805 Europeans, respectively. We performed univariate MR analysis using MR Egger, Weighted median (WM) and Inverse variance weighted (IVW) methods to assess causal effects among the three. Through the Genotype Tissue Expression (GTEx) database, single-nucleotide polymorphisms (SNPs) that affect the expression levels of T2DM-related genes were located using expression quantitative trait loci (eQTL). Results: MR analysis showed a significant causal relationship between T2DM and ED (WM, OR: 1.180, 95%CI: 1.010-1.378, P = 0.037; IVW, OR: 1.190, 95%CI: 1.084-1.300, P < 0.001). There is also a significant causal relationship between T2DM and MI (MR Egger, OR: 0.549, 95%CI: 0.317-0.952, P = 0.037; WM, OR: 0.593, 95%CI: 0.400, P = 0.010; IVW, OR: 0.767, 95%CI: 0.600-0.980, P = 0.034). ED may not cause MI (P > 0.05). We also found that rs6585827 corresponding to the PLEKHA1 gene associated with T2DM is an eQTL variant affecting the expression of this gene. Conclusion: T2DM has a direct causal effect on ED and MI. The level of PLEKHA1 expression suppressed by rs6585827 is potentially associated with a lower risk of T2DM.

Genomic Characterization and gE/gI-Deleted Strain Construction of Novel PRV Variants Isolated in Central China.

Pseudorabies virus (PRV) variants have caused substantial economic losses in the swine industry in China since 2011. To surveil the genetic variation in PRV field strains, here, two novel variant strains of PRV were isolated from Shanxi Province in central China and were designated SX1910 and SX1911. To identify the genetic characteristics of the two isolates, their complete genomes were sequenced, and phylogenetic analysis and sequence alignment revealed that field PRV variants have undergone genetic variations; notably, the protein-coding sequences UL5, UL36, US1 and IE180 exhibited extensive variation and contained one or more hypervariable regions. Furthermore, we also found that the glycoproteins gB and gD of the two isolates had some novel amino acid (aa) mutations. Importantly, most of these mutations were located on the surface of the protein molecule, according to protein structure model analysis. We constructed a mutant virus of SX1911 with deletion of the gE and gI genes via CRISPR/Cas9. When tested in mice, SX1911-ΔgE/gI-vaccinated mice were protected within a comparable range to Bartha-K61-vaccinated mice. Additionally, a higher dose of inactivated Bartha-K61 protected the mice from lethal SX1911 challenge, while a lower neutralization titer, higher viral load and more severe microscopic lesions were displayed in Bartha-K61-vaccinated mice. These findings highlight the need for continuous monitoring of PRV and novel vaccine development or vaccination program design for PRV control in China.

Frozen-thawed embryo transfer cycles in China: clinical outcomes of two and three multicellular embryos transfers.

PURPOSE: To analyze the clinical outcomes of frozen embryo transfer (FET) cycles when two or three multicellular embryos were transferred in Chinese women. METHODS: A retrospective study was conducted to analyze 980 FET cycles performed between January 2007 and October 2010. Two (785 cycles) or three (195 cycles) multicellular embryos were transferred. RESULTS: Both in patients under 35 years (n = 776) and those aged 35 to 39 years (n = 169), the transfer of two versus three multicellular embryos results in similar clinical pregnancy rates (CPR), implantation rates (IR) and live birth rates (LBR). In both age groups, the multiple pregnancy rate (MPR) was significantly higher in the three-embryo groups. Among women over 40 years of age (n = 35), there were no differences in the CPR, IR, MBR or LBR between the two groups CONCLUSIONS: Transferring two instead of three multicellular embryos in patients under 40 years old significantly decreases the risk of MPR without compromising PR, IR and LBR. In the age group above 40, transferring two instead of three multicellular embryos did not decrease PR, IR, MBR or LBR. Transferring more embryos when a patient had more unsuccessful cycles was not warranted in all patients.

The effect of human cumulus cells on the maturation and developmental potential of immature oocytes in ICSI cycles.

PURPOSE: To investigate the effect of human cumulus cells on the maturation and developmental potential of immature oocytes in ICSI cycles. METHODS: Immature oocytes were randomly divided into two groups: the cumulus-denuded oocyte group (group A) and the cumulus-intact oocyte group (group B). Only oocytes that reached metaphase II (MII) stage after in vitro maturation were used in the ICSI procedure. In vivo mature sibling MII oocytes served as the control group. Maturation rate, fertilization rate, embryo quality and developmental potential were examined. RESULTS: There was no significant difference in maturation rate between group A (68.16%) and group B (70.49%; P > 0.05). The total fertilization rate among the three groups was comparable (P > 0.05), while the zygotes with two pronuclei in group A (74.59%) or group B (75.97%) were significantly lower than those in control group (84.29%; P < 0.05). The available embryo rate in group A (11.49%) was markedly lower than that in group B (27.66%; P < 0.05), and both of them were significantly lower than that in control group (62.38%; P < 0.05). The proportion of ≥6-cell embryos in group B (45.74%) was notably higher than in group A (26.44%; P < 0.05), and both were markedly lower than in control group (65.92%; P < 0.05). The proportion of embryos with <10% fragmentation in group A (13.79%) was significantly lower than in group B (29.79%; P < 0.05), and both were notably lower than in control group (42.98%; P < 0.05). CONCLUSIONS: The presence of cumulus cells surrounding the immature oocytes during IVM before ICSI had no influence on nuclear maturation and fertilization, but leads to better subsequent embryonic development. This is perhaps mediated by an improvement in cytoplasmic maturation.