Effect of probiotics on children with autism spectrum disorders: a meta-analysis.

BACKGROUND: Researches have found that alteration of intestinal flora may be closely related to the development of autism spectrum disorder (ASD). However, whether probiotics supplementation has a protective effect on ASD remains controversial. This meta-analysis aimed to analyze the outcome of probiotics in the treatment of ASD children. METHODS: The Pubmed, Cochrane Library, Web of Science and Embase were searched until Sep 2022. Randomized controlled trials (RCTs) relevant to the probiotics and placebo treatment on ASD children were screened. Quality assessment of the included RCTs was evaluated by the Cochrane collaboration's tool. The primary outcomes were ASD assessment scales, including ABC (aberrant behavior checklist) and CBCL (child behavior checklist) for evaluating the behavior improvement, SRS (social responsiveness scale) for social assessment, DQ (developmental quotient) for physical and mental development and CGI-I (clinical global impression improvement) for overall improvement. The secondary outcome was total 6-GSI (gastrointestinal severity index). RESULTS: In total, 6 RCTs from 6 studies with 302 children were included in the systemic review. Total 6-GSI (MD=-0.59, 95%CI [-1.02,-0.17], P < 0.05) decreased significantly after oral administration of probiotics. Whereas, there was no statistical difference in ABC, CBCL, SRS, DQ and CGI-I between probiotics and placebo groups in ASD children. CONCLUSION: Probiotics treatment could improve gastrointestinal symptoms, but there was no significant improvement in ASD.

[Expression of Zfx in mouse testicular spermatogenic cells].

OBJECTIVE: To investigate the expression of Zfx gene in spermatogenic cells. METHODS: The testes of d6, d8, d17 and adult mice were collected, single cell suspension was prepared by combinatorial enzyme digestion, spermatogenic cells were isolated by BSA density gradient method, and Zfx expression was detected by real-time quantitative polymerase chain reaction (qRT-PCR) and Western Blot (WB). RESULTS: Single cell suspension prepared by combination enzyme digestion method and density gradient method laid with BSA can obtain various types of spermatogenic cells with purity>85%; The expression level of the Zfx gene is low in primitive type A spermatogonia, type A spermatogonia, and type B spermatogonia, whereas it is high in preleptotene spermatocytes, pachytene spermatocytes, and round spermatid cells. It is not expressed in elongating spermatids and mature sperm. CONCLUSION: Zfx gene exhibits periodic expression in various levels of spermatogenic cells and may be an important transcription factor involved in regulating meiosis in spermatogenic cells.

Decreased Event-Related Desynchronization of Mental Rotation Tasks in Young Tibetan Immigrants.

The present study aimed to explore the cortical activity underlying mental rotation in high-altitude immigrants via the event-related desynchronization (ERD), the electroencephalogram time-frequency analysis, and source localization based on electroencephalographic data. When compared with the low-altitude individuals, the reaction time of mental rotation tasks was significantly slower in immigrants who had lived in high-altitude areas for 3 years. The time-frequency analysis showed that the alpha ERD and the beta ERD within the time window (400-700 ms) were decreased during the mental rotation tasks in these immigrants. The decreased ERD was observed at the parietal-occipital regions within the alpha band and at the central-parietal regions within the beta band. The decreased ERD might embody the sensorimotor-related cortical activity from hypoxia, which might be involved in cognitive control function in high-altitude immigrants, which provided insights into the neural mechanism of spatial cognition change on aspect of embodied cognition due to high-altitude exposure.

Analysis of influencing factors related to elevated serum troponin I level for COVID-19 patients in Yichang, China.

BACKGROUND: Cardiac injury is a common condition among hospitalized coronavirus disease 2019 (COVID-19) patients, and is associated with a higher risk of mortality. However, the mechanism of myocardial injury in COVID-19 remains unclear. In this retrospective study, we compared the clinical characteristics of COVID-19 patients with different troponin I (TnI) levels during hospitalization to provide a clinical reference for the identification of those at high-risk. METHODS: In total, 218 patients diagnosed with COVID-19 in Yichang Central People's Hospital and Yichang Third People's Hospital between January 23 and February 19, 2020 were initially included. Of these patients, 89 underwent TnI testing during hospitalization and were finally included in the study. The medical history, clinical signs and symptoms at the time of admission, and laboratory test results were recorded. The patients were assigned to the normal TnI group (TnI <0.01 µg/L; n=67) or the elevated TnI group (TnI >0.01 µg/L; n=22). RESULTS: The incidence of elevated TnI in our patient cohort was 24.7%. There were significant differences between the two groups in the following factors: history of coronary heart disease (CHD), age, lymphocyte count, prothrombin time (PT), activated partial thromboplastin time (APTT), and levels of interleukin (IL)-6, C-reactive protein (CRP), myoglobin (MYO), lactate dehydrogenase (LDH), and albumin (all P<0.05). Binary logistic analysis showed that a history of CHD, age, lymphocyte count, IL-6, APTT, and MYO were influencing factors of elevated serum TnI. CONCLUSIONS: A history of CHD, advanced age, decreased lymphocyte count, increased IL-6, increased MYO, and prolonged APTT were independent influencing factors of elevated TnI in COVID-19 patients. COVID-19 patients with these characteristics are prone to myocardial injury.

Sex control by Zfy siRNA in the dairy cattle.

Zinc-finger Y is located in the short arm of the Y-chromosome and is a highly conserved gene that plays an important role in spermatogenesis. The objective of this study was to investigate the influence of silencing the Zfy gene during spermatogenesis on Y-sperm formation and offspring sex determination in Bos taurus cattle. Three recombinant expression vectors pLL3.7/a, pLL3.7/b and pLL3.7/c were evaluated and only pLL3.7/a effectively silenced the Zfy gene. The pLL3.7/a recombinant expression vector was injected into bull testes, using three injections. Semen was collected and preserved by extending and freezing. The frozen semen was subsequently used in artificial insemination of cows during a breeding season in accordance with the production plan on the farm where the experiment was conducted. Results showed that, after exposure to pLL3.7/a, sperm motility decreased (P < 0.01), but the sperm density was similar (p > 0.05) to the non-treated control semen. Injection of pLL3.7/a resulted in 72.0% female offspring, and was greater than the 49.4% female calves in the control (P ＜ 0.01), Results from this research suggests that the Zfy gene plays a role in the process of Y-sperm formation, and Zfy siRNA is a potential useful approach to control sex of offspring in cattle.

A genetic method for sex determination in Ovis spp. by interruption of the zinc finger protein, Y-linked (ZFY) gene on the Y chromosome.

The mammalian Y chromosome plays a critical role in spermatogenesis. However, the exact functions of each gene on the Y chromosome have not been completely elucidated, due, in part, to difficulties in gene targeting analysis of the Y chromosome. The zinc finger protein, Y-linked (ZFY) gene was first proposed to be a sex determination factor, although its function in spermatogenesis has recently been elucidated. Nevertheless, ZFY gene targeting analysis has not been performed to date. In the present study, RNA interference (RNAi) was used to generate ZFY-interrupted Hu sheep by injecting short hairpin RNA (shRNA) into round spermatids. The resulting spermatozoa exhibited abnormal sperm morphology, including spermatozoa without tails and others with head and tail abnormalities. Quantitative real-time polymerase chain reaction analysis showed that ZFY mRNA expression was decreased significantly in Hu sheep with interrupted ZFY compared with wild-type Hu sheep. The sex ratio of lambs also exhibited a bias towards females. Together, the experimental strategy and findings of the present study reveal that ZFY also functions in spermatogenesis in Hu sheep and facilitate the use of RNAi in the control of sex in Hu sheep.

Xeno-free culture of human spermatogonial stem cells supported by human embryonic stem cell-derived fibroblast-like cells.

Spermatogonial stem cells (SSCs) divide continuously to support spermatogenesis throughout postnatal life and transmit genetic information to the next generation. Here, we report the successful establishment of the method for the isolation and identification of human SSCs from testicular tissue, and to determine the culture conditions required to expand SSCs on human embryonic stem cell-derived fibroblast-like cells (hdFs). Large-scale cultures of SSCs were maintained on hdF feeder layers and expanded in the presence of a combination of cytokines and glial cell line-derived neurotrophic factor for at least 2 months. Cell surface marker analysis showed that SSCs retained high levels of alkaline phosphatase activity and stained strongly for anti-stage-specific embryonic antigen (SSEA)-1, OCT4 and CD49f. They also expressed the genes OCT4, SOX3 and STRA8 as detected by reverse transcription polymerase chain reaction (RT-PCR) analysis. These data clearly illustrate a novel approach for the growth of human SSCs using hdFs as feeder cells, potentially eliminating xenogeneic contaminants. This system provides a new opportunity for the study of the regulatory mechanism of the 'niche' that governs SSC self-renewal, and will be a valuable source of SSCs for potential clinical applications.

Leptin and varicocele-related spermatogenesis dysfunction: animal experiment and clinical study.

The objective of this study was to explore the relationships between varicocele-related spermatogenesis dysfunction and the expression of leptin and leptin receptors. In rats with experimental varicocele, the function of spermatogenesis, the expression of leptin and leptin receptors in testes were analysed; and in patients with varicocele-related male infertility, serum and seminal plasma levels of leptin, gonadal hormones and semen parameters were evaluated. In the testes of rats, leptin was expressed in seminiferous tubules and intersitium, leptin receptor was predominantly expressed in interstitium. The expression of leptin and its receptor in the testis of rats was not related to the weight of rat, but was inversely related to the weight of testis (r = -0.408, p = 0.009 and r = -0.433, p = 0.005, respectively), the Johnsen scores (r = -0.916, p = 0.000 and r = -0.863, p = 0.000, respectively), the seminiferous tubules diameter (r = -0.853, p = 0.000 and r = -0.870, p = 0.000, respectively) and the thickness of seminiferous epithelium (r = -0.929, p = 0.000 and r = -0.948, p = 0.000, respectively). In varicocele patients (N = 40), the sperm concentration and motility were significantly lower (p = 0.000) than those in the control group (N = 25), and the leptin level in seminal plasma was significantly higher (p = 0.000) than that in the control group. The leptin in serum and seminal plasma was positively related (r = 0.223, p = 0.002). The seminal plasma leptin level was inversely related to sperm concentration (r = -0.632, p = 0.000) and motility (r = -0.635, p = 0.000). There was no significant relation between serum leptin and seminal parameters and between leptin and gonadal hormone values. The dysfunction of spermatogenesis in varicocele-related infertile male is associated with increase in leptin and leptin receptors. Leptin may have local effects on the function of testis and spermatogenesis.

[The feeder layer of human embryonic fibroblasts supports the growth of human spermatogonial stem cells].

OBJECTIVE: To investigate the methods and conditions for the isolation, purification and culture of human spermatogonial stem cells (SSCs) on the feeder layer cells of human embryonic fibroblasts (hEFs). METHODS: SSCs isolated and purified from normal human fetal testicular tissues by sequential two-step enzyme digestion and Percoll uncontinuous density gradient centrifugation were cultured on the feeder layer cells of hEFs isolated from 5-9 weeks old human embryos. The surface markers SSEA-1 and OCT4 of the SSCs were detected by immunohistochemistry; the alkaline phosphatase (AKP) activity of the SSC clones measured; and the expressions of the SSC-related genes determined by RT-PCR. RESULTS: SSCs survived, proliferated and formed colonies on the feeder layers, and the colonies were highly positive for SSEA-1 and OCT4, with strong AKP activity and high expressions of the SSC-related genes. CONCLUSION: The feeder layer of hEFs supports the growth of human spermatogonial stem cells.

[Effects of growth hormone supplementation on erectile function and expression of nNOS in aging rats].

OBJECTIVE: To explore the effects of growth hormone( GH) supplementation on erectile function and expression of nNOS in the intracavernosal nerves in aging rats. METHODS: Twenty male Sprague-Dawley rats aged 18 months were randomly divided into Groups A and B, and ten 2-month-old male Sprague-Dawley rats included in Group C. 1 U/(kg x d) GH was given to Group A, and the same volume of saline to Groups B and C. After 8 weeks of treatment, evaluation was made of the erections induced by apomorphine (APO), maximal intracavernous pressure (ICP) induced by intracavernous papaverine injection and expression of nNOS in the intracavernosal nerves by streptavidin-peroxidase immunohistochemical techniques. RESULTS: After 8 weeks of treatment, the erection frequency, maximal ICP and expression of nNOS in the intracavernosal nerves of the rats in Groups A and C were significantly higher than those in Group B (P < 0.05 or P < 0.01). CONCLUSION: Growth hormone supplementation can improve the erectile function of aging rats, which may be attributed to the increase in the expression of nNOS in the intracavernosal nerves.

[Experimental research on human spermatozoa membrane proteins by two-dimensional gel electrophoresis].

OBJECTIVES: To analyze human spermatozoa membrane proteins by two-dimensional gel electrophoresis and to provide a basis for drawing the protein map of normal human spermatozoa membrane proteins. METHODS: Spermatozoa were purified by Percoll density centrifugation, and spermatozoa membrane proteins were analyzed by two-dimensional gel electrophoresis using isoelectric focusing and polyacrylamide gel electrophoresis. RESULTS: About 800 protein spots could be identified by the imaging analysis system. The molecular weight and isoelectric point of most proteins were 20,000 to 100,000 and 3.0 to 7.0 respectively. CONCLUSIONS: There are more than 800 types of proteins in the human spermatozoa membrane. The spermatozoa membrane protein can be identified with a certain precision by two-dimensional gel electrophoresis.

[Measurement of the reactive oxygen species and cytokines in the seminal plasma of leukocytospermic patients].

OBJECTIVES: To detect the levels of reactive oxygen species (ROS), superoxide dismutase(SOD) and interleukin 8(IL-8) in seminal plasma of infertile patients, and evaluate the possible relationship between those levels. METHODS: Semen was collected from normal donors (15 cases), infertile men without infection (16 cases), and infertile men with infection (leukocytospermia, 11 cases). The routine analysis of semen was accomplished, and then the levels of IL-8, malondialdehyde (MDA), SOD, and white blood cell (WBC) were examined. The correlative analysis between the level of ROS and other parameters in these populations was made. RESULTS: In leukocytospermic group, the levels of MDA, WBC, and IL-8 were higher than those in the other two groups (P < 0.001). Significantly positive correlation was observed between IL-8 and MDA (r = 0.852, P < 0.001) and between the levels of IL-8 and WBC. CONCLUSIONS: These findings suggest that increased oxidative stress in patients with leukocytospermia may cause the increase of IL-8(r = 0.818, P < 0.01). The increased oxidative stress may be due to defect in ROS scavenging system.

Cryopreservation-induced decrease in heat-shock protein 90 in human spermatozoa and its mechanism.

AIM: To study the protein changes of spermatozoa associated with sperm motility during sperm cryopreservation and its mechanism. METHODS: In 18 healthy men, the seminal sperm motility and HSP90 levels were studied before and after cryopreservation using SDS-PAGE, Western blotting and computerized image analysis. RESULTS: The sperm motility declined significantly after cryopreservation (P<0.01). The average grey level and the integrated grey level of sperm HSP90 before cooling were 34.1+/-3.2 and 243.0+/-21.6, respectively, while those after thawing were 23.2+/-2.5 and 105.7+/-28.5, respectively. Both parameters were decreased significantly (P<0.01). No HSP90 was found in the seminal plasma before and after cryopreservation. CONCLUSION: HSP90 in human spermatozoa was decreased substantially after cryopreservation. This may result from protein degradation, rather than leakage into the seminal plasma.

[Seminal plasma angiotensin II detection and its clinical implication].

OBJECTIVE: To investigate the variation of seminal plasma angiotension II (Ang II) in infertile men and its clinical implication. METHODS: Ang II values in paired blood plasma and seminal plasma from 43 infertile men(13 azoospermia, 8 asthenozoopermia, 17 asthenozoospermia and 5 cases with normal semen parameters) and 10 normal controls were obtained by SPE-HPLC-RIA. All semen samples with spermatozoa were analyzed by CASA for sperm count, motility and other parameters. Acrosome reaction rate (AR) was assessed by triple-stain. RESULTS: The mean concentration of seminal plasma Ang II was 4 times as high as that of blood plasma in all patients and controls (P < 0.01), but there was no correlation between them. The seminal plasma Ang II of azoospermic patients was higher than that of other infertile men and controls(P < 0.05), but no difference was found between the latter two groups. There was no correlation between seminal plasma Ang II values and other traditional parameters of sperm together with AR. CONCLUSIONS: Seminal plasma Ang II may be secreted locally in male reproductive tract. In addition to testis and epididymis, prostate and/or seminal vesicle may also be the source of it. The reason why seminal plasma Ang II of azoospermic patients is higher than that of others remains unknown. Further study is required to clarify the exact role of seminal plasma Ang II in the mechanisms of male fertility regulation.

[The effects of testosterone undecanoate on relaxation of rat corpus cavernosum smooth muscle in vitro].

OBJECTIVES: To investigate the role of Undecanoate (Andriol), as a kind of testosterone, in regulating the relaxation of isolated rat corpus cavernosum in vitro. METHODS: The castrated rats were given high and low dosage Andriol respectively, compared with intact and castrated rats. After treatment of 4 weeks, the corpora cavernosa were cut, trimmed as to strips. Norepinephrine(NE) was added to contract each of the tissue strips. Next, sodium nitroprusside(SNP), electric functional stimulation(EFS) were used to relax the strips. Percent relaxations were examined. RESULTS: High-dose Andriol(20 mg/kg) was significantly effective to castrated rats on percent relaxation induced by 10(-3) mol/L SNP and EFS. Low-dose Andriol(10 mg/kg) was also effective to relax strips induced by 10(-3) mol/L SNP. According to statistics, the differences were significant (P < 0.01). CONCLUSIONS: The percent relaxations of castrated rats were increased after taking Andriol, and it could increase the relaxation of corpara cavernosa.