The decline of fertility in male uremic patients is correlated with low expression of the cystic fibrosis transmembrane conductance regulator protein (CFTR) in human sperm.

BACKGROUND: The present study was designed to investigate the possible association between infertility of male uremic patients and expression of the cystic fibrosis transmembrane conductance regulator (CFTR) protein in their sperm. METHODS: Semen was collected and analyzed. Serum levels of FSH, LH and testosterone were measured by radioimmunoassay. The sperm CFTR expressions of 21 uremic patients and 15 renal transplant patients were measured and compared with those of 32 healthy and 33 infertile men. RESULTS: Only 9 ± 5.9% of sperm from uremic patients expressed CFTR, significantly less than those of the renal transplant patients (29 ± 14.3%, P< 0.001), the infertile men (42 ± 20.7%, P< 0.001) and the healthy men (51 ± 20.5%, P< 0.001). Furthermore, significantly fewer sperm from renal transplant patients expressed CFTR than those of the infertile men (P< 0.05) and the healthy men (P< 0.01). LH levels in uremic patients were significantly higher than in all other groups, whereas FSH levels in uremic patients were only significantly higher than in infertile and healthy men. There was no significant difference in testosterone level among the four categories. CONCLUSIONS: Sperm CFTR expression is depressed in uremic patients but recovers to some degree after renal transplant along with some improvement in fertility, indicating a 'reversible' change. These results suggest that the CFTR expression rate in sperm is correlated with the decline of uremic patients' fertility, and may be considered as a potential marker to assess the fertility of male uremic patients.

Human Umbilical Cord Mesenchymal Stem Cells to Treat Neuromyelitis Optica Spectrum Disorder (hUC-MSC-NMOSD): A Study Protocol for a Prospective, Multicenter, Randomized, Placebo-Controlled Clinical Trial.

Background: Neuromyelitis Optica spectrum disorder (NMOSD) is severe relapsing and disabling autoimmune disease of the central nervous system. Its optimal first-line treatment to reduce relapse rate and ameliorate neurological disability remains unclear. We will conduct a prospective, multicenter, randomized, placebo-controlled clinical trial to study the safety and effectiveness of human umbilical cord mesenchymal stem cells (hUC-MSCs) in treating NMOSD. Methods: The trial is planned to recruit 430 AQP4-IgG seropositive NMOSD patients. It consists of three consecutive stages. The first stage will be carried out in the leading center only and aims to evaluate the safety of hUC-MSCs. Patients will be treated with three different doses of hUC-MSCs: 1, 2, or 5 × 106 MSC/kg·weight for the low-, medium-, and high-dose group, respectively. The second and third stages will be carried out in six centers. The second stage aims to find the optimal dosage. Patients will be 1:1:1:1 randomized into the low-, medium-, high-dose group and the controlled group. The third stage aims to evaluate the effectiveness. Patients will be 1:1 randomized into the optimal dose and the controlled group. The primary endpoint is the first recurrent time and secondary endpoints are the recurrent times, EDSS scores, MRI lesion numbers, OSIS scores, Hauser walking index, and SF-36 scores. Endpoint events and side effects will be evaluated every 3 months for 2 years. Discussion: Although hUC-MSC has shown promising treatment effects of NMOSD in preclinical studies, there is still a lack of well-designed clinical trials to evaluate the safety and effectiveness of hUC-MSC among NMOSD patients. As far as we know, this trial will be the first one to systematically demonstrate the clinical safety and efficacy of hUC-MSC in treating NMOSD and might be able to determine the optimal dose of hUC-MSC for NMOSD patients. Trial registration: The study was registered with the Chinese Clinical Trial Registry (CHICTR.org.cn) on 2 March 2016 (registration No. ChiCTR-INR-16008037), and the revised trial protocol (Protocol version 1.2.1) was released on 16 March 2020.

[Impregnate occasion for male renal transplant recipients].

OBJECTIVE: To explore the impregnate occasion for male renal transplant recipients. METHODS: Twenty-six male renal transplant recipients were divided into 3 groups according to the post-transplantation time and the administered dose of Cyclosporine A: 7 in Group A (less than 6 months after renal transplantation and at the dose of 4.1-6 mg/[kg x d]), 11 in Group B (6-24 months and 2.1-4 mg/[kg x d]) and 8 in Group C (longer than 24 months and 1.3-2 mg/[kg x d]). The semen of the patients were detected and compared with those of 12 normal volunteers. RESULTS: Statistically significant differences were observed in sperm motility and sperm head deformity between Group A and C (P < 0.05), but not in pH value and sperm volume, vitality and concentration among the 3 groups (P > 0.05). CONCLUSION: For male renal transplant recipients, 2 years or longer after the transplantation is the most suitable time for impregnation.

[Sperm morphology does not influence fertilization rate in vitro].

OBJECTIVE: To determine the correlation of the rate of morphologically normal sperm, sperm head defect, teratozoosperm index (TZI) and sperm deformity index (SDI) with the fertilization outcome of IVF and intracytoplasmic sperm injection (ICSI). METHODS: We detected normal sperm morphology, sperm head defect, TZI and SDI by Krüger's strict criteria and compared them with the rate of fertilization. RESULTS: In either IVF or ICSI, the fertilization rate did not show any correlation with the normal sperm morphology rate, the head defect rate, TZI and SDI, nor any statistic differences were found in the rates of fertilization, clinical pregnancy and quality embryos either between the SDI > 1.6 and the SDI < 1.6 groups (P > 0.05) or between the normal sperm morphology rate > 15% and < 15% groups (P > 0.05). CONCLUSION: Sperm morphology has no predictive value for the rate of fertilization in vitro.

Morphological characteristics of spermatozoa before and after renal transplantation.

AIM: To investigate the changes of the spermatozoa ultrastructures before and after renal transplantation in uremic patients. METHODS: The sperm of five uremic patients before and after transplantation and four healthy volunteers were collected and examined by scanning electron microscopy. RESULTS: Abnormal spermatozoa were found in patients pre-transplantation; abnormalities included deletion of the acrosome, absence of the postacrosomal and postnuclear ring, dumbbell-like changes of the head, tail curling, and absence of the mitochondrial sheath in the mid-segment. After renal transplantation, most of the spermatozoa became normal. CONCLUSION: There are many abnormalities with regard to the appearance and structure of the head, acrosome, mitochondria and tail of the spermatozoa in uremic patients. The majority of the spermatozoa returned to normal after renal transplantation, but a few still presented some abnormalities possibly relating to the administration of immunosuppressants.

[Effects of different dosages of cyclosporine A on the semen parameters of renal transplant patients].

OBJECTIVES: To evaluate the effects of different dosages of cyclosporine A (CsA) on the main semen parameters and sperm morphology of the patients after renal transplantation. METHODS: The semen of 18 patients after renal transplantation treated with different dosages of CsA was analyzed and the semen parameters and sperm morphology were compared with those of 12 normal volunteers. RESULTS: There was not significant difference between the main parameters of the patients treated with 1.5-3.0 mg.Kg-1.d-1 of CsA and 3.1-5.5 mg.Kg-1.d-1 of CsA and those of the volunteers (P > 0.05), but the rate of normal sperm morphology was significantly different between the two groups(P < 0.05). CONCLUSIONS: Different therapeutic dosages of CsA did not have any effect on most of the semen parameters of the patients after renal transplantation, but did affect the sperm morphology.

Marital status and fertility of 185 male renal transplant recipients in China.

A questionnaire was designed to assess the effects of renal transplantation in men of reproductive age on marital status and fertility. The study sought to correlate recipients' marital status and fertility with the health of the recipients after the transplantation, the health of children they fathered after the procedure, and the functioning of the transplanted kidney. Male recipients (n = 243) who were single and of reproductive age before renal transplantation were selected from 2007 recipients of a renal transplant recorded in the authors' hospitals in China. Of the 243 surveyed, 185 completed the questionnaire and participated in follow-up in the clinic or by telephone. Their marital status and fertility were investigated. Of the 185 recipients, 69 got married 12-88 months (mean, 32.19 +/- 14.30 months) after renal transplantation, and 62 of 69 couples were actively attempting to become pregnant. Fifty-three patients fathered 54 children, including 1 pair of twins, 9-72 months (mean, 25.81 +/- 15.33 months) after marriage. The birth weights of the newborns ranged from 2500 to 4600 g (mean, 3395 +/- 456.80 g). These children developed well. Nine patients did not father any children, and 3 of these 9 cases were attributable to infertility in the wife. Seven patients were using contraceptives. Three recipients suffered from chronic graft rejection and resumed hemodialysis 2-11 years after they fathered children. In addition, 2 patients died after fathering 1 child: 1 from dysfunction of the transplanted kidney 9 years after birth of his child, and another in an accident 1 year after his child's birth. Our findings suggest that, like men without renal transplants, male recipients of renal transplants can get married and father children, and the transplantation procedure appears to have no significant effect on the children fathered afterwards, on the recipients' health, or on the functioning of the transplanted kidney. It is very important to indicate that, in addition to needing contraception if they do not conceive, male renal transplant recipients should expect fertility rates that are similar to those of the general population.

[Effects of different segments of acupuncture serum on eosinophil counts in the rat with eosinophilia].

OBJECTIVE: To further analyze and identify effective components of anti-asthma in acupuncture serum. METHODS: Changes of eosinophils in the peripheral blood of rats with eosinophilia were observed for 10 days after intravenous injection of the different segments of serum (serum: normal saline = 1:20, 2.5 mL/kg, from the first day of the model establishment, for 3 consecutive days). RESULTS: After intravenous injection of different segments of serum, the eosinophil counts in the peripheral blood decreased significantly from the 3rd day as compared with those of the model group (P<0.05). CONCLUSION: The effective components of acupuncture serum from asthmatic rats treated by acupuncture for eosinophils are not a single component, and acupuncture stimulation may produce many kinds of components of anti-asthma.

Montelukast, a cysteinyl leukotriene receptor-1 antagonist, dose- and time-dependently protects against focal cerebral ischemia in mice.

Our previous studies showed that cysteinyl leukotriene receptor-1 (CysLT1) antagonist pranlukast has a neuroprotective effect on cerebral ischemia in rats and mice. However, whether the neuroprotective effect of pranlukast is its special action or a common action of CysLT1 receptor antagonists remains to be clarified. This study was performed to determine whether montelukast, another CysLT1 receptor antagonist, has the neuroprotective effect on focal cerebral ischemia in mice, and to observe its dose- and time-dependent properties. Permanent focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO). Montelukast was injected intraperitoneally either as multiple doses (once a day for 3 days and 30 min before MCAO) or as a single dose (at 30 min before, 30 min after, or 1 h after MCAO), respectively, and pranlukast and edaravone were used as controls. The neurological deficits, infarct volumes, brain edema, neuron density, and Evans blue extravasation in the brain were determined 24 h after MCAO. Pretreatments with multiple doses or a single dose of montelukast (0.1 and 1.0 mg/kg) before MCAO significantly attenuated all the ischemic insults. Post-treatment with a single dose of montelukast (0.1 and 1.0 mg/kg) at 30 min after MCAO also significantly decreased brain edema and infarct volume, but not neurological deficits. However, post-treatment with a single dose of montelukast at 1 h after MCAO had no significant effect. Pranlukast showed the same effects as montelukast, but edaravone attenuated the ischemic insults only with multiple doses before MCAO. Thus, montelukast has a dose- and time-dependent neuroprotective effect on permanent focal cerebral ischemia in mice, with an effective dose range of 0.1-1.0 mg/kg and a therapeutic window of 30 min. These findings further support the therapeutic potential of CysLT1 receptor antagonists in the treatment of cerebral ischemia at earlier phases.

[Expression and purification of recombinant huwentoxin-I in Pichia pastoris].

HWTX-I is a peptide neurotoxin purified from the crude venom of the Chinese bird Spider Selenocosmia Huwena, which has analyesic activity. rHWTX-I expressed by P. pastoris and secreted to culture supernatant was first precipitated by (NH4)2SO4, then it was isolated and desalted by ultrofiltration following by ion exchange chromatography of CM column, after reverse phase HPLC of C18 column and vacuum drying, the pure HWTX-I protein was obtained which was proved to be recombinant HWTX-I by Tricine SDS-PAGE, MALDI-TOF mass spectrometry, amino acid composition analysis, the N-terminal amino acid sequence and its biological activity. The final yield of the purified HWTX-I was about 80 mg/L accounting for 23.6% of its total secretory proteins.

Wwp2, an E3 ubiquitin ligase that targets transcription factor Oct-4 for ubiquitination.

The POU transcription factor Oct-4 is a master regulator affecting the fate of pluripotent embryonic stem cells. However, the precise mechanisms by which the activation and expression of Oct-4 are regulated still remain to be elucidated. We describe here a novel murine ubiquitin ligase, Wwp2, that specifically interacts with Oct-4 and promotes its ubiquitination both in vivo and in vitro. Remarkably, the expression of a catalytically inactive point mutant of Wwp2 abolishes Oct-4 ubiquitination. Moreover, Wwp2 promotes Oct-4 degradation in the presence of overexpressed ubiquitin. The degradation is blocked by treatment with proteasome inhibitor. Fusion of a single ubiquitin to Oct-4 inactivates its transcriptional activity in a heterologous Oct-4-driven reporter system. Furthermore, overexpression of Wwp2 in embryonic stem cells significantly reduces the Oct-4-transcriptional activities. Collectively, we demonstrate for the first time that Oct-4 can be post-translationally modified by ubiquitination and that this modification dramatically suppresses its transcriptional activity. These results reveal that the functional status of Oct-4, in addition to its expression level, dictates its transcriptional activity, and the results open up a new avenue to understand how Oct-4 defines the fate of embryonic stem cells.