Circ-LRP1B functions as a competing endogenous RNA to regulate proliferation, apoptosis and oxidative stress of LPS-induced human C28/I2 chondrocytes.

Circular RNAs (circRNAs) are crucial for the pathogenesis of human diseases, including osteoarthritis (OA). Here, we set to elucidate the biological action of circ-LRP1B in OA pathogenesis. Human C28/I2 chondrocytes were stimulated by lipopolysaccharide (LPS). Circ-LRP1B, nuclear factor, erythroid 2 like 1 (NRF1) and microRNA (miR)-34a-5p were quantified by quantitative real-time PCR (qRT-PCR) or immunoblotting. Cell viability, proliferation, and apoptosis abilities were gauged by MTT, 5-Ethynyl-2'-Deoxyuridine (EdU) staining, and flow cytometry assays, respectively. Direct relationship between miR-34a-5p and circ-LRP1B or NRF1 was validated by RNA pull-down and dual-luciferase reporter assays. Circ-LRP1B was found to be underexpressed in OA cartilage and LPS-stimulated C28/I2 chondrocytes. Enforced expression of circ-LRP1B promoted cell proliferation, and repressed apoptosis and oxidative stress, as well as impacted OA-specific hallmarks expression of LPS-stimulated C28/I2 cells. NRF1 was identified as a downstream effector of circ-LRP1B function. Moreover, NRF1 was identified as a miR-34a-5p target in LPS-stimulated C28/I2 cells. Circ-LRP1B acted as a competing endogenous RNA (ceRNA) for miR-34a-5p to involve the post-transcriptional regulation of NRF1 expression. Furthermore, the effects of circ-LRP1B overexpression partly depended on the reduction of available miR-34a-5p. These findings demonstrate that circ-LRP1B functions as a ceRNA to regulate the proliferation, apoptosis and oxidative stress of LPS-stimulated human C28/I2 chondrocytes by miR-34a-5p/NRF1 network.

Analysis of Related Influencing Factors of Deep Vein Thrombosis after Lumbar Internal Fixation and Treatment Strategy.

Lumbar internal fixation is a traditional surgical method for the treatment of degenerative diseases of the lumbar spine. However, due to its large surgical trauma, it easily causes complications such as deep venous thrombosis (DVT) after the operation. DVT refers to the abnormal coagulation of blood in deep veins, blocking the lumen, causing venous blood return disorder, causing venous blood return disorder to cause swelling and pain, which affects the recovery of the patient's lumbar spine function. In severe cases, even complicated pulmonary embolism endangers the life and health of the patient. Therefore, it is extremely important to explore the related influencing factors and effective treatment of DVT. The purpose of this study was to investigate the influencing factors and effective treatment of DVT after lumbar internal fixation. Univariate analysis and multivariate a logistic regression model were used to analyze the related factors affecting DVT after lumbar internal fixation. Conventional treatments such as anticoagulation, promotion of venous blood return, and improvement of limb circulation were given to patients with DVT, and functional exercise was guided to compare the hypercoagulability and hyperviscosity of blood in patients with DVT before and after treatment. The results showed that the incidence of DVT after lumbar internal fixation was related to age, BMI, and bed time. Getting out of bed for functional exercise in time after surgery can effectively prevent the formation of DVT. Preoperative grading examination, intraoperative intervention, postoperative physical exercise, and other preventive guidance can be carried out according to different individuals during the perioperative period to prevent the formation of postoperative DVT.

Development and Validation of a Machine Learning Algorithm for Predicting Response to Anticholinergic Medications for Overactive Bladder Syndrome.

OBJECTIVE: To develop and externally validate a prediction model for anticholinergic response in patients with overactive bladder (OAB). METHODS: A machine learning model to predict the likelihood of anticholinergic treatment failure was constructed using a retrospective data set (n=559) of female patients with OAB who were treated with anticholinergic medications between January 2010 and December 2017. Treatment failure was defined as less than 50% improvement in frequency, urgency, incontinence episodes, and nocturia, and the patient's subjective impression of symptomatic relief. Patients were stratified by age (younger than 40 years, 40-60 years, and older than 60 years), and number of previously failed medications. K-fold stratified cross-validation was performed on each stratum using machine learning algorithms. Of these, the random forest model was the most accurate. This model was refined using internal cross validation within each stratum. The area under the curve (AUC) was calculated for each stratum and used to identify the optimal operating points for prediction of treatment failure. The random forest model was then externally validated using a prospectively collected data set (n=82) of women treated with anticholinergic medications at a different clinical site between January 2018 and December 2018. RESULTS: The global accuracy of the final model was 80.3% (95% CI 79.1-81.3), and the AUC was 0.77 (95% CI 0.74-0.79). Using the external validation data set, the model's sensitivity and specificity was 80.4% (95% CI 66.5-89.7%) and 77.4% (95% CI 58.6-89.7%), respectively. The model performed best in women aged younger than 40 years (AUC 0.84, 95% CI 0.81-0.84) and worst in women aged older than 60 years who had previously failed medication (AUC 0.71, 95% CI 0.67-0.75). CONCLUSION: Our externally validated machine learning prediction model can predict anticholinergic treatment failure during the standard 3-month treatment trial period with greater than 80% accuracy. The model can be accessed at https://oabweb.herokuapp.com/app/pre/.

Feasibility Study of Tissue Transglutaminase for Self-Catalytic Cross-Linking of Self-Assembled Collagen Fibril Hydrogel and Its Promising Application in Wound Healing Promotion.

Collagen-based bio-hydrogels are undoubtedly a hot spot in the development of biological dressings for wound healing promotion. Herein, glutamine transaminase (TGase), a biological nontoxic cross-linker with high specific activity and reaction rate under mild conditions, was utilized for the self-catalytic cross-linking of the regenerated collagen (COL) fibril hydrogel fabricated through a molecular self-assembly method. The results showed that the natural triple helical conformation of COL remained completely integrated after self-catalytic cross-linking TGase, which was definitively the fundamental for maintaining its superior bioactivity. It was worth noting that TGase could promote the self-assembly process of COL building blocks into a higher order D-period cross-striated structure. Also, the reconstructed TGase cross-linked COL fibrils exhibited a higher degree of interfiber entanglements with more straight and longer fibrils. Meanwhile, the thermal stability of COL was significantly improved after introducing TGase. Besides, the cytocompatibility analysis suggested that the regenerated COL fibril hydrogel showed excellent cell growth activity and proliferation ability when the dosage of TGase is less than 40 U/g. Further, animal experiments indicated that the targeted COL fibril hydrogel could significantly promote skin wound healing, exhibiting better capacity of skin tissue for regeneration than the COL hydrogel untreated as expected. Therefore, the reconstructed TGase cross-linked COL fibril hydrogel could serve as a novel soft material for wound healing promotion.

[Diagnosis and treatment of idiopathic hypogonadotropic hypogonadism in males].

Idiopathic hypogonadotropic hypogonadism (IHH) is often caused by hyposecretion of gonadotropin and consequently affects male fertility. The patient with IHH has a smaller penis and testes with spermatogenic dysfunction. At present, IHH is treated mainly with hCG, hMG, GnRH, and their different combinations. However, due to the lack of large-sample evidence, it is not yet clear which therapy is the best option. This article presents an overview of our experience in the treatment of IHH in the last decade and a review of relevant literature, aiming at a deeper insight into this male disease.

The angiogenesis in decellularized scaffold-mediated the renal regeneration.

There are increasing numbers of patients underwent partial nephrectomy, and recovery of disturbed renal function is imperative post partial nephrectomy. We previously have demonstrated the decellularized (DC) scaffolds could mediate the residual kidney regeneration and thus improve disturbed renal function after partial nephrectomy. However, the cellular changes including the angiogenesis in the implanted DC scaffold has not yet been elaborated. In this study, we observed that the scaffold promoted the proliferation of human umbilical vein endothelial cells (HUVEC) that adhered to the DC scaffold in vitro. We next examined the pathological changes of the implanted DC graft in vivo, and found a decreased volume of the scaffold and a dramatic angiogenesis within the scaffold. The average microvessel density (aMVD) increased at the early stage, while decreased at the later stage post transplantation. Expression level of vascular endothelial growth factor (VEGF) showed similar dynamic changes. In addition, many endothelial cells (ECs) and endothelial progenitor cells (EPCs) were distributed in the region which contained active angiogenesis in the scaffold. However, the implanted graft became fibrosis and the angiogenesis degraded at final stage roughly 8 weeks post transplantation. Our data indicate that DC scaffold can be vascularized in vivo and possible mechanisms are discussed.

Association between polymorphisms in the serotonin 2C receptor gene and premature ejaculation in Han Chinese subjects.

INTRODUCTION: Lifelong premature ejaculation (LPE) is characterized by persistently shorter intravaginal ejaculation latency time (IELT) than found acceptable by the patient or his partner. It has been postulated to be a neurobiological dysfunction with genetic vulnerability and is related to disturbances of central serotonin (5-hydroxytryptamine, 5-HT) neurotransmission and 5-HT receptor function. AIM: To investigate the relationship between the C-759T and G-697C polymorphisms of the 5-HT(2C) receptor and LPE. METHODS: A prospective study was conducted in 106 Han Chinese men with LPE, characterized by IELT of less than 1 min, and 84 healthy controls with IELT of more than 3 min. All subjects were genotyped for the C-759T and G-697C polymorphisms located in the promoter region of the 5-HT(2C) receptor. The frequencies of genotypes and single nucleotide mutations were compared between the two groups. RESULTS: Three genotypes were detected both in the men with LPE and in the control group: -759C/-697G, -759T/-697C, and -759C/ -697C. Genotype -759T/-697G was not detected. The frequency of genotype -759T/-697C was higher in patients with LPE than in the control group (30.2 vs. 11.9%, p < 0.05), whereas the frequency of genotype -759C/-697G was lower in patients with LPE than in the control group (66.0 vs. 83.3%, p < 0.05). No difference was found for genotype -759C/ -697C between the two groups. Mutations at -759T and -697C were more frequent in patients than in the control group (-759T: 30.2 vs. 13.3%, p < 0.05; -697C: 30.4 vs. 16.7%, p < 0.05, respectively). CONCLUSIONS: Our findings indicated that polymorphisms in the 5-HT(2C) receptor gene are associated with LPE, and men who carry the -759T or -697C genotype have increased odds of premature ejaculation. Further investigation in this field is necessary.

[Experience of using three phosphodiesterase type 5 inhibitors in the same period for the treatment of erectile dysfunction].

OBJECTIVE: To observe and compare the effects and adverse events of the three phosphodiesterase type 5 (PDE-5) inhibitors sildenafil, vardenafil and tadalafil used in the same period for the treatment of erectile dysfunction (ED). METHODS: A total of 311 ED patients were treated with the three PDE-5 inhibitors in the same period, 134 with sildenafil, 109 with vardenafil and 88 with tadalafil, and followed up for 2-11 months. Their effects were compared. RESULTS: Sildenafil produced satisfactory effect in 72 patients (79.12%), vardenafil in 63 (81.81%) and tadalafil in 52 (78.78%), respectively. And the efficacy rates of sildenafil, vardenafil and tadalafil in the treatment of ED with premature ejaculation were 55.56%, 44.44% and 42.3%, respectively. Our study also showed that the young and the newly married preferred vardenafil, the middle-aged chose tadalafil and the older and long-term users tend to take sildenafil. The main reasons for discontinuing the use of the three inhibitors were their high price, incomplete effect and the patients' worry about their adverse action. CONCLUSION: The three PDE-5 inhibitors can produce basically similar effects on ED, all with safety and few mild adverse events.

The effect of alpha-adrenergic antagonists in chronic prostatitis/chronic pelvic pain syndrome: a meta-analysis of randomized controlled trials.

The effectiveness of alpha-adrenergic antagonists on patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has not been supported by well-evaluated study. The meta-analysis was performed to supply the best evidence about use of this class of drugs in CP/CPPS. A fully recursive literature search to June 2005 was conducted in PubMed, EMBASE, the Cochrane Controlled Trials Register, and the Chinese Biomedicine Database to identify potentially relevant randomized controlled trials. RevMan4.2 was used for statistical analysis. Nine studies with 734 patients were included. Combined analysis showed a significant reduction of total NIH-CPSI or I-PSS in patients with treatment duration of more than 3 months. There were also valuable results in urinary symptom alleviation. Alpha-adrenergic antagonists did not show benefit in pain. The meta-analysis revealed that the use of alpha-adrenergic antagonists was warranted in CP/CPPS, and the treatment duration should be long enough (more than 3 months).

Preliminary study of the relationship between DAZ gene copy deletions and spermatogenic impairment in Chinese men.

Copy deletion screening of DAZ gene family on the Y chromosome in 485 patients with idiopathic azoospermia or oligozoospermia and 236 fertile men revealed that the prevalence of deletion patterns of the entire DAZ gene and DAZ1/DAZ2 gene were significantly higher in the patients than in fertile men. The deletion patterns correlate with spermatogenic impairment in a Chinese population.

DAZ1/DAZ2 cluster deletion mediated by gr/gr recombination per se may not be sufficient for spermatogenesis impairment: a study of Chinese normozoospermic men.

AIM: To explore the possible effect of the deleted in azoospermia (DAZ) copy cluster deletion on spermatogenesis in the Chinese population, the deletion of the azoospermia factor c (AZFc) region was analyzed in 346 normozoospermic men. METHODS: Three DAZ single nucleotide variant loci and seven AZFc-specific sequence-tagged sites were examined with polymerase chain reaction (PCR)-restriction fragment length polymorphism and routine PCR. RESULTS: Five (1.4%) of the normozoospermic men were found to have deletion of gr/gr-DAZ1/DAZ2. None of the men were found to have b2/b4-entire DAZ deletion. CONCLUSION: The presence of gr/gr-DAZ1/DAZ2 deletion in five men with normozoospermia suggests that this deletion per se may not be sufficient for spermatogenic impairment in Chinese men.

[Progress in the study of stem Leydig cells].

Leydig cells are the primary source of testosterone. As is true for all specialized cells, Leydig cells originally arise from primitive undifferentiated stem cells, stem Leydig cells (SLCs). The huge potential of SLCs is to be used therapeutically to restore testosterone levels in males with androgen deficiency due to diverse causes. The initial paradigm for cell therapy would call for harvesting the SLCs of an androgen deficient male, amplifying these cells in vitro, inducing differentiation in vitro, and then implanting the mature Leydig cells back into the same individual.

[Important genetic etiology of infertile Chinese males: chromosome abnormality and deletion of DAZ gene copy in the AZFc region of Y chromosome].

OBJECTIVE: To investigate the correlation of male infertility with abnormality of chromosomal quantity and construction and with the deletion of DAZ gene copy in the AZFc region of Y chromosome. METHODS: Included in the study were 247 azoospermic and 206 severe oligozoospermic patients, as well as 210 fertile men as controls. Multi-PCR and PCR-RFLP were used to analyze the deletion of DAZ gene copies in the AZFc region of Y chromosome. Chromosomal quantity and construction were detected by G-band in the 453 patients. RESULTS: In the azoospermic and severe oligozoospermic patients, the incidences of chromosomal abnormality were 12.6% and 8.3%; the rates of complete DAZ deletion were 7.7% and 11.2%, and the rates of DAZ1/DAZ2 deletion were 7.3% and 4.9% respectively, but no deletion was detected in the controls. CONCLUSION: There is a high frequency of chromosomal abnormality and DAZ gene copy deletion in patients with azoospermia and oligospermia, which suggests that chromosomal abnormality and partial and complete deletion of DAZ gene copy might be important genetic causes of Chinese male infertility.

[Semen quality and sperm morphologic study of infertile men with varicocele].

OBJECTIVE: To examine the semen quality and the sperm morphology in infertile men with varicocele. METHODS: Semen from 98 infertile men with varicocele were studied and those of 130 normal semen donors were taken as the control. Semen analysis was performed based on the methods described in the WHO manual and sperm morphology was evaluated by WHO criteria. RESULTS: A significantly reduced percentage of normal morphologic sperm and of forward progression were found in patients with varicocele comparing with those of the control (P <0.001). The head defects were observed as the predominant type of sperm malformation. CONCLUSION: The varicocele increases malformed sperm in ejaculates, which may result from impaired male fertility by varicocele. Sperm morphologic assessment with WHO criteria provides a sensitive and practical measurement of sperm damage in infertile men with varicocele.

[Clinical study of sildenafil in the treatment of premature ejaculation complicated by erectile dysfunction].

OBJECTIVES: To evaluate the efficacy and safety of sildenafil citrate in the treatment of premature ejaculation (PE) complicated by erectile dysfunction (ED). METHODS: Forty-five patients of PE complicated by ED received flexible doses of sildenafil from 50 to 100 mg for 1 to 3 months. Intravaginal ejaculatory latency time (IELT) and sexual satisfaction ratio (SSR) of partner were recorded to evaluate the effect of PE treatment, as well as the general efficacy and satisfaction of ED treatment. And the difference of IIEF-5 before and after the treatment were compared. RESULTS: Twenty-seven patients had their PE improved and the effective rate was 60%. Forty patients reported the improvement in erection and the percentage of erectile improvement was 88.88%. All the 27 patients with improvement of PE achieved effective erection through the administration of 50 mg sildenafil and the satisfaction rate reached 81.48%. On the other hand, only 1 case (5.56%) reported satisfaction over the treatment in the 18 patients who did not obtain improvement of PE. Between the PE improvement group and non-improvement group, there were significant differences (P < 0.001) shown in IIEF-5 scores before and after the treatment. Mild or moderate side effects were reported in 9 patients(20%), who recovered without any treatment. CONCLUSIONS: To premature ejaculation patients with ED, sildenafil can safely and effectively improve their erectile function, the satisfaction over the ED treatment outcome means that their PE symptoms could be alleviated.

Novel mutations of PKD1 gene in Chinese patients with autosomal dominant polycystic kidney disease.

BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is a common disease in China. The major gene responsible for ADPKD, PKD1, has been fully characterized and shown to encode an integral membrane protein, polycystin 1, which is thought to be involved in cell-cell and cell-matrix interaction. Until now, 82 mutations of PKD1 gene have been reported in European, American, and Asian populations. However, there has been no report on mutations of the PKD1 gene in a Chinese population. METHODS: Eighty Chinese patients in 60 families with ADPKD were screened for mutations in the 3' region of the PKD1 gene using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and DNA-sequencing techniques. RESULTS: Three mutations were found. The first mutation is a 12593delA frameshift mutation in exon 45, and the polycystin change is 4129WfsX4197, 107 amino acids shorter than the normal polycystin (4302aa). The second mutation is a 12470InsA frameshift mutation in exon 45, producing 4088DfsX4156, and the predicted protein is 148 amino acids shorter than the normal. The third one is a 11151C-->T transition in exon 37 converting Pro3648 to Leu. In addition, nine DNA variants, including IVS44delG, were identified. CONCLUSIONS: Three mutations in Chinese ADPKD patients are described and all of them are de novo mutations. Data obtained from mutation analysis also suggests that the mutation rate of the 3' single-copy region of PKD1 in Chinese ADPKD patients is very low, and there are no mutation hot spots in the PKD1 gene. Mutations found in Chinese ADPKD patients, including nucleotide substitution and minor frameshift, are similar to the findings reported by other researchers. Many mutations of the PKD1 gene probably exist in the duplicated region, promoter region, and the introns of PKD1.