[Sperm-hyaluronan binding assay in in vitro fertilization and embryo transfer].

OBJECTIVE: To explore the application value of sperm-hyaluronan binding (SHB) assay in in vitro fertilization and embryo transfer (IVF-ET). METHODS: This retrospective study included 163 cases of IVF-ET performed in our hospital from January to August 2019 due to female fallopian tube abnormality-induced infertility. The men were found with normal seminal parameters at semen analysis. According to the SHB rate, we divided the patients into a normal group (n = 126) and an abnormal group (n = 37) and analyzed the general conditions and the rates of normal fertilization, cleavage, available embryos, embryo implantation and clinical pregnancy. RESULTS: Statistically significant differences were observed between the normal and abnormal groups in the rates of SHB, normal fertilization and available embryos (P < 0.01 or P < 0.05), but not in the general conditions or the rates of cleavage, embryo implantation and clinical pregnancy. CONCLUSIONS: The SHB rate is not correlated with sperm concentration or the percentages of progressively motile sperm and morphologically normal sperm. Sperm-hyaluronan binding assay helps predict the outcomes of IVF-ET and embryo quality. An SHB rate of <58.5% indicates low rates of normal fertilization and available embryos, but has no significant correlation with the rates of embryo implantation and clinical pregnancy.

[Design and exploration of genetic comprehensive experiments based on Ds insertion mutants].

The cultivation of innovative abilities has become an important guide for higher education in China. Strengthening the integrated knowledge to design experiments is an effective way to improve undergraduate students' innovative abilities. Herein we designed a comprehensive experiment for molecular genetics by utilizing a rice Ds insertion mutant identified previously in our research project. In the comprehensive experiment, we adopt the method of scientific research as the main line of teaching and take the interesting phenotype of the rice mutant as the breakthrough point to reform and innovate genetics laboratory teaching. On the basis of this, we combined the progressive teaching method and guided the students to learn the TAIL-PCR skill and conduct an innovative experiment through expanding their knowledge. The comprehensive experiment will deepen students' understandings of the relationship between genotypes and phenotypes, help them master the effective way of thinking and technologies for scientific research to further improve their ability of the integrated application capability of theory and practices.

Abnormal expression of TLRs may play a role in lower embryo quality of women with polycystic ovary syndrome.

Toll-like receptors (TLRs) localize in mammalian ovary, including granulosa cells, cumulus cells, and theca cells. Previous studies demonstrated that TLRs may be important for the cumulus-oocyte complex expansion and fertilization. There is no evidence to indicate that the deletion of TLRs will induce infertility; however, the abnormal expression of TLRs may decrease oocyte quality and fertility rate. In the present study, we investigated the effects of polycystic ovary syndrome (PCOS) on the expression of TLRs in cumulus cells by using western-blot and quantitative real-time PCR (qRT-PCR) analyses. We found that the expression of TLR4 and 9 in cumulus cells was influenced significantly by PCOS. We also observed that overweight/obesity changed the expression of TLR2 and 5 in cumulus cells of PCOS subjects. In addition, we found that the rate of available embryos of women with PCOS was slightly lower. These results indicate that the abnormal expression of TLRs in cumulus may be a reason for the lower embryo quality of women with PCOS. ABBREVIATIONS: ART: assisted reproductive technology BMI: body mass index COC: cumulus-cell-oocyte complex PCOS: polycystic ovary syndrome q RT-PCR: quantitative real-time PCR TLRs: Toll-like receptors.

[Effects of tillage and straw returning on microorganism quantity, enzyme activities in soils and grain yield].

A two-year field study with split plot design was conducted to investigate the effects of different soil tillage (conventional tillage, CT; deep tillage, DT; subsoil tillage, ST) and straw returning (all straw retention, AS; no straw returning, NS) on microorganism quantity, enzyme activities in soil and grain yield. The results showed that, deep or subsoil tillage and straw returning not only reduced the soil bulk density and promoted the content of organic carbon in soil, but increased the soil microbial quantity, soil enzyme activities and grain yield. Furthermore, such influences in maize season were greater than that in wheat season. Compared with CT+NS, DT+AS and ST+AS decreased the soil bulk density at 20-30 cm depth by 8.5% and 6.6%, increased the content of soil organic carbon by 14.8% and 12.4%, increased the microorganism quantity by 45.9% and 33.9%, increased the soil enzyme activities by 34.1% and 25.5%, increased the grain yield by 18.0% and 19.3%, respectively. No significant difference was observed between DT+AS and ST+AS. We concluded that retaining crop residue and deep or subsoil tillage improved soil microorganism quantity, enzyme activities and crop yield.

Analysis on genetic diversification and heterosis in autotetraploid rice.

Polyploidization has played an important role in plant evolution and is a pathway for plants to increase genetic diversification and to get higher heterosis comparing with that of diploid does. This study was undertaken to assess the genetic variation and relationships among 40 autotetraploid rice genotypes and their counterpart diploid cultivars with 99 SSR markers screened from published rice genome. The 99 SSR markers detected polymorphism among autotetraploid genotypes and revealed a total of 291 alleles with an average of 2.949 alleles per locus. Autotetraploid lines showed higher genetic diversity and significant variation in agronomic traits than diploid cultivars. Phylogenetic analysis revealed that most of autotetraploid lines were genetically different from their diploid parents, and inter-subspecific hybrids were prepared on the basis of genetic distance between parents. Inter-subspecific autotetraploid hybrids showed a higher and positive heterobeltiosis and competitive heterosis than diploid hybrids, especially for grain yield. Genetic distance appeared not to predict heterosis in diploid rice for all traits; however, it showed a significant correlation with grain yield, grain length and grain length to width ratio in autotetraploid rice. This extensive research on autotetraploid heterosis and genetic diversity will be useful for the development of autotetraploid rice hybrids.

[Outcomes of ICSI with sperm from different sources: a retrospective study of 431 cycles].

OBJECTIVE: To compare the outcomes of intracytoplasmic sperm injection (ICSI) with retrieved epididymal and testicular sperm for obstructive azoospermia and with ejaculated sperm for severe oligozoospermia and asthenospermia. METHODS: We retrospectively analyzed 431 ICSI cycles, which were divided according to sperm sources into Groups A (n=287 in patients with severe oligozoospermia or asthenospermia using ejaculated sperm), B (n=109 in obstructive azoospermia patients with sperm retrieved by percutaneous epididymal sperm aspiration, PESA) and C (n=35 in obstructive azoospermia patients with sperm retrieved by testicular sperm extraction, TESE). Comparisons were made among the three groups in the rates of embryo implantation, fertilization, pregnancy, cleavage, and miscarriage. RESULTS: Group A showed statistically significant differences from Groups B and C in the rates of embryo implantation and pregnancy (18.46% vs. 25.23% and 28.76%, 31.23% vs. 42.16% and 39.39%, P < 0.05). But no significant differences were seen in the rates of fertilization, cleavage and miscarriage among the three groups (P > 0.05). CONCLUSION: The rates of embryo implantation and clinical pregnancy are higher in patients with obstructive azoospermia than in those with severe oligozoospermia or asthenospermia after ICSI with ejaculated sperm.

[The culture and identification of rat testis Leydig cell].

OBJECTIVE: To establish a primary culture method of rat testis Leydig cell. METHODS: The primary rat Leydig cells were treated with or without 4 U/ml human chorionic gonadotropin (hCG), and testosterone in culture medium was detected by radioimmunoassay. The morphology and biological characteristics of Leydig cell were observed. RESULTS: The culture cells were highly homogeneous, proliferative and had a high differentiation rate. The high purified Leydig cells were verified by their dynamic morphological changes and 3beta-hydroxysteroid dehydrogenase delta4-delta5 isomerase (3beta-HSD) histochemical staining. The testosterone secretion induced by hCG significantly increased (P < 0.05) 24 hours after inoculation than that induced without hCG in the control. CONCLUSION: It suggests that the Leydig cell cultured in vitro may secrete high concentration of testosterone, and this study laid the basis of androgen replacement therapy for partial androgen deficiency in aging male.

[Surgical therapy for azoospermia with ejaculatory duct obstruction].

OBJECTIVE: To investigate the effect of transurethral resection of ejaculatory ducts (TURED) for azoospermia with ejaculatory duct obstruction (EDO). METHODS: From June 2003 to December 2004, 20 azoospermia with EDO were diagnosed, diagnostic criteria included a history, physical examination, semen analyses, semen fructose measurement, endocrine assessment, testicular biopsy and transrectal ultrasonography (TRUS); All 20 cases were treated by TURED. Fifteen of them were followed up more than 3 months after the treatment. The semen samples of them were analysed at 3-month intervals in post-therapy. RESULTS: Semen analyses in all 20 cases showed the typical characteristics of EDO, low semen volume (0.4-1.6 ml), azoospermia, low pH, absent or low semen fructose. TRUS showed the main etiology factor of EDO was a midline cyst in 11, lateral cystic lesions in 2, the remaining 7 cases had dilated ejaculatory duct with or without dilated seminal vesicles. Among 15 cases followed up more than 3 months after TURED, 10/15 (67%) had an improvement in semen parameters and 3/15 (20%) had pregnancies. Semen analyses had not been done in anther 5 cases. CONCLUSION: Transurethral resection of ejaculatory ducts may be a safe and effective method for the treatment of azoospermia with EDO.

[Experimental study of changes in microscopic characteristics and ability of secreting testosterone of aged SD rat leydig cells].

OBJECTIVE: To research the changes in microscopic characteristics and ability of secreting testosterone between aged SD rat Leydig cells and young SD rat Leydig cells. METHODS: The total and free serum testosterone levels of serum both young and aged rats were examined. The changes in microscopic characteristics between young and aged rat Leydig cells were observed under microscope and electron microscope. The testosterone secreted by cultured Leydig cells of stimulated by hCG and Forskolin both groups were detected. RESULTS: A significant difference was found in both total and free testosterone levels between young and old rats (P < 0.05). Aged SD rat Leydig cells were observed smaller in volume and more markedly stained than young ones; The secretion ability of aged rat Leydig cells was found lower than that of young rat Leydig cells with or without hCG and Forskolin stimulation (P < 0.05). CONCLUSION: The secreting index of aged SD rat Leydig cells is lower than that of young rat Leydig cells both in vivo and vitro, and the reason is the system of synthesizing testosterone is arrested.

[Culture and identification of human adult Leydig cells].

OBJECTIVE: To obtain a simple and effective method to isolate and purify adult Leydig cells. METHODS: The testes of human adults were digested and then the density gradient centrifugation of the cells was performed with four different Percoll concentrations (60%, 34%, 26%, 21%) to isolate Leydig cells, whose characteristics were identified by cytological observation staining, 3beta-HSD staining and detection of hCG and testosterone secretion. RESULTS: High-concentration (> 90%) purified Leydig cells were acquired, and many identification experiments demonstrated the adequate testosterone secretory function of the isolated and purified Leydig cells. CONCLUSION: This method is easy and efficient for the isolation and purification of adult Leydig cells.

[Measures for optimizing the efficacy of sildenafil on patients with erectile dysfunction].

Incorrect drug administration, lack of sexual stimulation, the way of obtaining sildenafil and associating risk factors of erectile dysfunction like severe cardiovascular or neurogenic diseases were the common causes of initial sidenafil failure. To improve the therapeutic efficacy for patients with initial sidenafil failure, education about how to use sildenafil properly plays an important role. Moreover, dose titration, more attempts, a good follow-up and sildenafil taking on a daily basis will yield a high response rate to sildenafil rechallenge.

Two types of new ferritin cDNA sequences from Xenopus laevis germinal vesicle oocytes.

Some of Xenopus ferritin cDNA family genes have already been sequenced. In this study, we report that two ferritin cDNA genes have been cloned from the Xenopus laevis germinal vesicle (GV) oocytes. The deduced proteins have different lengths with varied sequences when compared with the published Xenopus ferritins. One of them is the ferritin light chain homologous (LCH), which is reported for the first time in Xenopus and the other is the ferritin heavy chain homologous (HCH) that is first reported in Xenopus GV oocyte.