Dissecting peri-implantation development using cultured human embryos and embryo-like assembloids.

Studies of cultured embryos have provided insights into human peri-implantation development. However, detailed knowledge of peri-implantation lineage development as well as underlying mechanisms remains obscure. Using 3D-cultured human embryos, herein we report a complete cell atlas of the early post-implantation lineages and decipher cellular composition and gene signatures of the epiblast and hypoblast derivatives. In addition, we develop an embryo-like assembloid (E-assembloid) by assembling naive hESCs and extraembryonic cells. Using human embryos and E-assembloids, we reveal that WNT, BMP and Nodal signaling pathways synergistically, but functionally differently, orchestrate human peri-implantation lineage development. Specially, we dissect mechanisms underlying extraembryonic mesoderm and extraembryonic endoderm specifications. Finally, an improved E-assembloid is developed to recapitulate the epiblast and hypoblast development and tissue architectures in the pre-gastrulation human embryo. Our findings provide insights into human peri-implantation development, and the E-assembloid offers a useful model to disentangle cellular behaviors and signaling interactions that drive human embryogenesis.

Effects of the body mass index of males on hormone levels, sperm and embryo parameters, and clinical outcomes in non-obstructive azoospermia: a systematic review and meta-analysis.

Background: Research has shown that the body mass index (BMI) is not correlated with sperm retrieval outcomes, while serum testosterone and gonadotropins are related to the BMI in non-obstructive azoospermia (NOA). Previously, no comprehensive assessment had been conducted on the effect of the BMI of males in NOA. This study sought to comprehensively evaluate the effects of the BMI of males on hormone levels, sperm and embryo parameters, and clinical outcomes in NOA. Methods: The PubMed, Embase, Cochrane Library, and Web of Science databases were searched to retrieved relevant articles published up to May 27, 2022. Articles which examined patients with NOA (population), included patients with a BMI ≥25 kg/m2 (intervention) versus patients with a BMI <25 kg/m2 (comparator), assessed reproductive hormones, sperm and embryo parameters, and clinical outcomes (outcome), and were cohort studies (study design) were included. The quality of the included studies was assessed by the Newcastle-Ottawa Scale (NOS). A sensitivity analysis was conducted for all the outcomes. Results: A total of 12 studies comprising 2,994 NOA patients were included. Patients with a BMI ≥25 kg/m2 had lower follicle-stimulating hormone (FSH) [pooled weighted mean difference (WMD): -0.67, 95% confidence interval (CI): -0.94 to -0.41, P<0.001] and total testosterone (TT) (pooled WMD: -1.35, 95% CI: -2.10 to -0.60, P<0.001) levels than those with a normal weight (BMI <25 kg/m2). The testicular volume of the BMI ≥25 kg/m2 group was larger than that of the normal weight group (pooled WMD: 0.26, 95% CI: 0.09 to 0.44, P=0.003). The average BMI of the group with successful sperm extraction was lower than that of the group with failed sperm extraction (pooled WMD: -0.97, 95% CI: -1.89 to -0.04, P=0.041). The live-birth rate of the BMI ≥25 kg/m2 group was lower than that of the normal weight group [pooled relative risk (RR) =0.88, 95% CI: 0.78 to 0.99, P=0.031]. Conclusions: The BMI of the males was an important factor affecting the FSH and TT levels, testicular volume, sperm retrieval success, and live-birth rate in NOA. Weight management may benefit NOA patients.

LH level on ovulation trigger day has a different impact on the outcomes of agonist and antagonist regimens during in vitro fertilization.

BACKGROUND: To assess the impact of the luteinizing hormone level on ovulation trigger day (LHOTD) on in vitro fertilization (IVF) outcomes in gonadotropin-releasing hormone (GnRH) agonist and antagonist regimens during fresh embryo transfer cycles. METHODS: A stepwise, progressive multivariate regression model was introduced to assess the effect of the LHOTD on clinical pregnancy and live birth rates. Mantel‒Haenszel stratification analysis was used to examine the association between the LHOTD and clinical outcomes with the antagonist regimen. RESULTS: The LHOTD had different distributions in the agonist and antagonist regimens. The cycles were assigned into three LHOTD tertile groups. In the agonist regimen, compared with the 1st tertile (T1), in the 2nd (T2) and 3rd (T3) tertiles, the adjusted odds ratios (ORs) (95% confidence intervals [CIs], P value) were 1.187 (1.047-1.345, 0.007) and 1.420 (1.252-1.610, < 0.001) for clinical pregnancy, respectively, and 1.149 (1.009-1.309, 0.036) and 1.476 (1.296-1.681, < 0.001) for live birth. In the antagonist regimen, there was no significant difference in clinical pregnancy and live birth rates among the tertiles. However, in the stratified group of patients aged less than 35 years, the ORs (95% CIs, P value) of T2 and T3 were 1.316 (1.051-1.648, 0.017) and 1.354 (1.077-1.703, 0.009) for clinical pregnancy, respectively, and 1.275 (1.008-1.611, 0.043) and1.269 (0.999-1.611, 0.051) for live birth. Moreover, there was a discrepancy in the results among the subdivided LHOTD T1 groups adopting the antagonist regimen. Compared with that of the < 1.06 mIU/mL subgroup, the ORs (95% CIs, P value) of the > 1.5 mIU/mL subgroup were 1.693 (1.194-2.400, 0.003) for clinical pregnancy and 1.532 (1.057-2.220, 0.024) for live birth after eliminating potential confounders. CONCLUSIONS: The LHOTD was profoundly suppressed in the agonist regimen, and its level was positively correlated with clinical pregnancy and live birth rates. In contrast, in the flexible antagonist regimen, the LHOTD was significantly higher than that in the agonist regimen and did not correlate with the outcome, except for women in the nonadvanced age group and those with an excessively suppressed LHOTD. Further investigation is required to determine the rationale for these findings.

[Expression of the kallistatin gene in rat corpus cavernosum].

OBJECTIVE: To investigate the expression of the kallistatin gene in the corpus cavernosum and search for some new molecular targets for the regulation of penile erectile function and treatment of ED. METHODS: Using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), Western blot and immunofluorescence staining, we detected the expression of kallistatin in the rat corpus cavernosum and compared it with that in the aorta. RESULTS: The results of RT-qPCR and Western blot revealed both mRNA and protein expressions of kallistatin in the rat corpus cavernosal tissue, with no statistically significant difference from those in the aorta (P > 0.05). Immunofluorescence staining showed that kallistatin was expressed in both endothelial and smooth muscle cells in the corpus cavernosum and localized in the cytoplasm, with no statistically significant difference from its expression in the aorta (P > 0.05) either. CONCLUSIONS: The kallistatin gene is highly expressed in the corpus cavernosum and localized in cavernosal endothelial and smooth muscle cells, suggestive of its involvement in the cellular function of cavernosal endothelial and smooth muscle cells and its participation in the regulation of penile erectile function.

Association between the RAGE (receptor for advanced glycation end-products) -374T/A gene polymorphism and diabetic retinopathy in T2DM

Summary Objective: Interaction between advanced glycation end-products (AGEs) and receptor for AGEs (RAGE) in cells could affect both extracellular and intracellular structure and function, which plays a pivotal role in diabetic microvascular complications. The results from previous epidemiological studies on the association between RAGE gene -374T/A polymorphism and diabetic retinopathy (DR) risk were inconsistent. Thus, we conducted this meta-analysis to summarize the possible association between RAGE -374T/A polymorphism and DR risk. Method: We searched all relevant articles on the association between RAGE -374T/A polymorphism and DR risk from PubMed, Cochrane Library, ScienceDirect, Wanfang, VIP and Chinese National Knowledge Infrastructure (CNKI) web databases up to August 2016. Odds ratio (OR) with 95% confidence interval (CI) were calculated to assess those associations. All analyses were performed using the Review Manager software. Results: Nine case-control studies, including 1,705 DR cases and 2,236 controls were enrolled, and the results showed that the A allele of RAGE -374T/A polymorphism was significantly associated with increased DR risk in dominant model (TA/AA vs. TT: OR=1.22, 95CI 1.05-1.41, p=0.006) and heterozygote model (TA vs. TT: OR=1.26, 95CI 1.07-1.47, p=0.005). The subgroup analysis by ethnicity showed that significantly increased DR risk was found in both Asian and Caucasian populations. Conclusion: This meta-analysis reveals that the A allele of RAGE -374T/A polymorphism probably increase DR risk.

Association between the RAGE (receptor for advanced glycation end-products) -374T/A gene polymorphism and diabetic retinopathy in T2DM.

OBJECTIVE: Interaction between advanced glycation end-products (AGEs) and receptor for AGEs (RAGE) in cells could affect both extracellular and intracellular structure and function, which plays a pivotal role in diabetic microvascular complications. The results from previous epidemiological studies on the association between RAGE gene -374T/A polymorphism and diabetic retinopathy (DR) risk were inconsistent. Thus, we conducted this meta-analysis to summarize the possible association between RAGE -374T/A polymorphism and DR risk. METHOD: We searched all relevant articles on the association between RAGE -374T/A polymorphism and DR risk from PubMed, Cochrane Library, ScienceDirect, Wanfang, VIP and Chinese National Knowledge Infrastructure (CNKI) web databases up to August 2016. Odds ratio (OR) with 95% confidence interval (CI) were calculated to assess those associations. All analyses were performed using the Review Manager software. RESULTS: Nine case-control studies, including 1,705 DR cases and 2,236 controls were enrolled, and the results showed that the A allele of RAGE -374T/A polymorphism was significantly associated with increased DR risk in dominant model (TA/AA vs. TT: OR=1.22, 95CI 1.05-1.41, p=0.006) and heterozygote model (TA vs. TT: OR=1.26, 95CI 1.07-1.47, p=0.005). The subgroup analysis by ethnicity showed that significantly increased DR risk was found in both Asian and Caucasian populations. CONCLUSION: This meta-analysis reveals that the A allele of RAGE -374T/A polymorphism probably increase DR risk.

Anti-tumor effects of fusion vaccine prepared by renal cell carcinoma 786-O cell line and peripheral blood dendritic cells of healthy volunteers in vitro and in human immune reconstituted SCID mice.

Dendritic cells (DC), as professional antigen presenting cells, play the central role in the process of body initiating the anti-tumor immunity, and the study on DC anti-tumor vaccine has become heated in recent years. In this study, we used polyethylene glycol (PEG) to induce renal cell carcinoma (RCC) 786-O cell line fused with peripheral blood DC of healthy volunteers, and discuss the biological characteristics of fusion vaccine and its anti-tumor effects in vitro and in human immune reconstituted SCID mice model of RCC. The study found that PEG could effectively induce cell fusion, and the expressions of CD86 and HLA-DR in fusion vaccine group were significantly up-regulated compared with the DC control group; the secretion of IL-12 was much higher and longer than that of the control; the functions of dendritic cell-tumor fusion vaccine to stimulate the proliferation of allogenic T lymphocytes and to kill RCC786-O cells in vitro were significantly higher than those of the control group, and after the killing, apoptosis body was observed in the target cells; after the injection of fusion vaccine into human immune reconstituted SCID mice model of RCC786-O via vena caudalis, the volume of mice tumor was reduced significantly, proliferation index of tumor cells decreased obviously compared with that of the control group, and more hemorrhage and putrescence focuses presented, accompanying large quantity of lymphocytes soakage. The results of this experimental study shows that fusion vaccine of RCC786-O cell line and DC can significantly stimulate the proliferation of allogenic T cells and specifically inhibit and kill RCC cells in vitro and in vivo, which makes the DC-RCC786-O fusion vaccine a possible new way of effective RCC immunotherapy.