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Bioactive terpenoids from Curcuma wenyujin (CW), as specialized metabolites, have huge medicinal properties. Accumulating evidence suggested that Jasmonates (JAs) and abscisic acid (ABA) shake hands and coordinately control a wide range of plant growth and defense activities, including the production of specialized metabolites. However, knowledge of how JA and ABA signalings interact to regulate terpenoid biosynthesis (TBS) is limited. In this study, integrative transcriptome and metabolome analyses revealed that exogenous methyl jasmonate (MeJA) and ABA activate the biosynthesis of terpenoids by upregulating the expressions of genes related to the TBS pathway, JA and ABA responses. Most of the differentially changed terpenoids and transcripts were commonly induced by MeJA and ABA. Interactive networks of "gene-gene/metabolite" indicated that CwbHLH2 , a MeJA and ABA dual-responsive regulator, is tightly related to terpenoid production. Furthermore, transgenic experiments revealed that overexpression and RNA interference of CwbHLH2 upregulated and downregulated the contents of representative terpenoids in the MeJA- and ABA-treated transgenic hairy roots, respectively, with 1.52–3.06-fold increases and 29.6–68.7% reduction. Importantly, the genetic transformation of CwbHLH2 enhanced the expressions of multiple TBS pathway genes and bidirectionally enlarged the JA and ABA signalings caused by MeJA or ABA, demonstrating that CwbHLH2 plays a critical interrelational role in integrating MeJA- and ABA-induced TBS regulation. Altogether, our results not only advance a new view of the crosstalk between JA and ABA in modulating TBS but also provide a powerful strategy for improving the yield of bioactive terpenoids in CW. [Display omitted] • MeJA and ABA significantly activate the accumulation of terpenoids in C. wenyujin leaves. • MeJA and ABA coregulate wide transcriptional changes to steer terpenoid formation. • CwbHLH2 plays a critical interrelational role in integrating JA and ABA signalings. • CwbHLH2 promotes MeJA- and ABA-inducible defense responses. • CwbHLH2 provides a powerful tool for improving the yield of terpenoids in Curcuma. [ABSTRACT FROM AUTHOR]

Due to the versatility of load bearing and heat insulation, foam-filled composite corrugated sandwich plates have a broad application in aerospace industry as thermal insulation structures for hypersonic vehicles. In this paper, a C 0 higher-order layerwise finite element model is presented for aeroelastic analysis of foam-filled composite corrugated sandwich plates in supersonic flow with equivalent properties. The homogenization method for the foam-filled corrugated core is formulated by force and energy equivalence under different assumptions of force or moment distributions. In the proposed layerwise model, the composite sandwich plate is divided into three sections through thickness, while a higher-order displacement field is assumed for the single-layer orthotropic core section and a first-order displacement field for the two face-sheet sections made of multi-layer laminates. The effect of transverse normal strain is also taken into consideration. The unsteady aerodynamic pressure is evaluated by the first-order Piston theory. An eight-noded isoparametric element with 16 degrees of freedom per node is used for finite element method. The accuracy of the present model is verified with existing results in the literature. The influence of geometric parameters and material properties on critical dynamic pressure are discussed. [ABSTRACT FROM AUTHOR]

A parametric variational principle and the corresponding numerical algorithm are proposed to solve a linear-quadratic (LQ) optimal control problem with control inequality constraints. Based on the parametric variational principle, this control problem is transformed into a set of Hamiltonian canonical equations coupled with the linear complementarity equations, which are solved by a linear complementarity solver in the discrete-time domain. The costate variable information is also evaluated by the proposed method. The parametric variational algorithm proposed in this paper is suitable for both time-invariant and time-varying systems. Two numerical examples are used to test the validity of the proposed method. The proposed algorithm is used to astrodynamics to solve a practical optimal control problem for rendezvousing spacecrafts with a finite low thrust. The numerical simulations show that the parametric variational algorithm is effective for LQ optimal control problems with control inequality constraints. [ABSTRACT FROM AUTHOR]

The anchorage performances and affecting factors of resin holts were studied by the combination method of theoretical analysis, laboratory tests, numerical modeling and field measurement. The stress distribution along bolts in pullout and fully anchored states was theoretically analyzed; the pullout tests of holts with different profiles and anchored in boreholes with various diameter were carried out in laboratory, and the affecting degree of bolt profiles and diameter difference between holts and boreholes to the bolt pullout force was obtained; the pullout force of bolts anchored in simulated boreholes with different temperature and water dripping content was measured, and the relations between pullout force and temperature, water dripping content were got. By means of FLAC3D,the shear stresses in resin and surrounding rock along bolts with different profiles, resin annulus thickness, egocentricity, and anchored in rocks of different strength were calculated, the affecting of these factors to shear stress distribution was analyzed, and stress distribution features were obtained. The soft rock anchorage test was carried out in underground, in the Fengshui-gou Coal Mine, the Pingzhuang coal mining district, and the pullout loads of bolts and cables in soft rock with different water content were measured. Finally, the suggestions for improving resin abchorage performances and enhancing anchorage force were put forward. [ABSTRACT FROM AUTHOR]

Abstract: 1,1-Dichloro-2,2 bis(p-chlorophenyl) ethylene (p,p′-DDE), the major metabolite of 2,2-bis(4-chlorophenyl)-1,1,1-trichloroethane (DDT), is a known persistent organic pollutant and male reproductive toxicant. It has antiandrogenic effect. However, the mechanism by which p,p′-DDE exposure causes male reproductive toxicity remains unknown. To elucidate the mechanism underpinning the testicular effects of p,p′-DDE, we sought to investigate Fas/FasL apoptotic pathway in the testis of prepubertal rats, including Fas, FasL, caspase-8, -3, and NF-κB. Animals were administered with different doses of p,p′-DDE (0, 20, 60, 100mg/kg b.wt) every other day by intraperitoneal injection for 10 days. The results indicated that p,p′-DDE exposure at over 20mg/kg b.wt showed the induction of apoptotic cell death. p,p′-DDE could induce increase in the MDA level, and decrease in SOD and GSH-Px activity. Significant elevations in the mRNA levels of Fas along with an increase in FasL, caspase-3, -8 were observed in 100mg/kg b.wt group. In protein level, p,p′-DDE could induce increase of FasL and reduction of procaspase-8. NF-κB p65 was activated by p,p′-DDE treatment in rat testis. In addition, the activities of caspase-3, -8 were increased in 100mg/kg b.wt group. Taken together, these results lead us to speculate that in vivo exposure to p,p′-DDE might induce testicular apoptosis in prepubertal rats through the Fas/FasL pathway. [Copyright &y& Elsevier]

• A novel viroid-like RNA (324 nt) was discovered in Dendrobium TSA. • Its infectivity and circularity were proven in D. officinal. • Tentatively designated as Dendrobium viroid (DVd). • DVd has a predicted rod-like secondary structure containing a CCR and a TCR. • DVd groups in the genus Apscaviroid and shares 56% nucleotide identity with CVd V. • DVd had a low incidence (0.96%) in Dendrobium species under natural conditions. Viroids are small circular RNA molecules which have been found to infect many dicot species. Only coconut cadang-cadang viroid and coconut tinangaja viroid have been reported so far to infect a monocot (coconut). Data mining in silico has proven an efficient approach to identify new viruses/viroids, and a systematic screen of public transcriptomic data revealed a 648 nucleotides (nt) sequence potentially representing a novel viroid-like RNA in a transcriptome shotgun assembly from Dendrobium officinale. This sequence contained two central conserved regions (CCRs) characteristic of members of the genus Apscaviroid , indicating that the viroid-like RNA is 324 nt in length. The infectivity of dimeric RNA transcripts generated by in vitro transcription of a synthetic cDNA, was demonstrated by directly injecting into the stems of young Dendrobium officinale plants. The presence of this novel viroid, tentatively designated as Dendrobium viroid (DVd), in the inoculated plants was confirmed by 2D-PAGE together with northern hybridization. DVd is predicted to have a rod-like secondary structure containing a CCR and a terminal conserved region (TCR), and phylogenetic analysis shows that it groups with the known members of the genus Apscaviroid. It is most closely related to citrus viroid V (56% nt identity). A field survey revealed a low DVd incidence (0.96%) in Dendrobium species in China. To our best knowledge, DVd is the only viroid known to infect orchids and the third one from monocotyledonous plants. [ABSTRACT FROM AUTHOR]

Background: Antibody-drug conjugates (ADCs) targeting the RON receptor, a tumorigenic factor contributing to cancer malignancy, has been considered as a novel strategy for cancer therapy. Here we describe a humanized antibody recognizing the RON plexin-semaphorin-integrin (PSI) domain with increased drug delivery capability for potential clinical application. Method: Monoclonal antibody PCM5B14 specific to the human and monkey RON PSI domain was generated and characterized by various immunological methods. Humanized antibody H5B14 was created by grafting PCM5B14 complementarity-determining regions into human IgG1/κ acceptor frameworks and conjugated with monomethyl auristatin E and duocarmycin to form two H5B14-based ADCs. Stability of H5B14-based ADCs in human plasma was measured using hydrophobic interaction chromatography. Various biochemical and biological assays were used to determine ADC- regulated RON internalization, cell viability, spheroid formation, and death of cancer stem-like cells. Efficacies of H5B14-based ADCs in vivo were validated using tumor xenograft models. Maximal tolerated doses of H5B14-based ADCs were established in mice. Results: H5B14 was highly specific to the human RON PSI domain and superior over other anti-RON ADCs in induction of RON internalization in various cancer cell lines tested. H5B14-based ADCS had a drug to antibody ratio of ~ 3.70:1 and were stable in human plasma with a minimal dissociation within a 10-day period. Functionally, H5B14-mediated drug delivery decreased cell viability at early stages with an average IC50 at ~ 20 nM in multiple cancer cell lines examined. H5B14-based ADCs also inhibited spheroid formation and caused death of cancer stem-like cells with RON+/CD44+/ESA+ phenotypes. In vivo, H5B14-based ADCs in a single injection inhibited tumor xenograft growth mediated by multiple cancer cell lines. Tumoristatic concentrations calculated from xenograft tumor models were in the range of 0.63 to 2.0 mg/kg bodyweight. Significantly, H5B14-based ADCs were capable of eradicating tumors at variable levels across multiple xenograft models regardless their malignant statuses. Toxicologically, H5B14-based ADCs were well tolerated in mice up to 60 mg/kg. Conclusion: H5B14-based ADCs targeting the RON PSI domain are superior in inducing RON internalization, leading to robust drug delivery and overall inhibition and eradication of tumors in multiple xenograft models. These findings warrant H5B14-based ADCs for clinical trials in the future. [ABSTRACT FROM AUTHOR]

Following publication of the original article [1], the author reported two errors in the authors affiliations: [ABSTRACT FROM AUTHOR]

Background: Aberrant expression of the RON receptor tyrosine kinase is a pathogenic feature and a validated drug target in various types of cancers. Currently, therapeutic antibodies targeting RON for cancer therapy are under intensive evaluation. Here we report the development and validation of a novel humanized anti-RON antibody-drug conjugate for cancer therapy. Methods: Antibody humanization was achieved by grafting sequences of complementarity-determining regions from mouse monoclonal antibody Zt/g4 into human IgG1/κ acceptor frameworks. The selected humanized Zt/g4 subclone H1L3 was conjugated with monomethyl auristatin E using a dipeptide linker to form H-Zt/g4-MMAE. Pharmacokinetic analysis of H-Zt/g4-MMAE was determined using hydrophobic interaction chromatography and a MMAE ADC ELISA kit. Biochemical and biological assays were used for measuring RON expression, internalization, cell viability and death. Therapeutic efficacies of H-Zt/g4-MMAE were validated in vivo using three pancreatic cancer xenograft models. Toxicological activities of H-Zt/g4-MMAE were determined in mouse and cynomolgus monkey. Results: H-Zt/g4-MMAE had a drug to antibody ratio of 3.77:1 and was highly stable in human plasma with a dissociation rate less than 5% within a 20 day period. H-Zt/g4-MMAE displayed a favorable pharmacokinetic profile in both mouse and cynomolgus monkey. In vitro, H-Zt/g4-MMAE induced RON internalization, which results in killing of pancreatic cancer cells with IC50 values at 10–20 nM. In vivo, H-Zt/g4-MMAE inhibited pancreatic cancer xenograft growth with tumoristatic concentrations at 1~3 mg/kg bodyweight. Significantly, H-Zt/g4-MMAE eradicated tumors across multiple xenograft models regardless their chemoresistant and metastatic statuses. Moreover, H-Zt/g4-MMAE inhibited and eradicated xenografts mediated by pancreatic cancer stem-like cells and by primary cells from patient-derived tumors. Toxicologically, H-Zt/g4-MMAE is well tolerated in mice up to 60 mg/kg. In cynomolgus monkey, H-Zt/g4-MMAE up to 30 mg/kg had a manageable and reversible toxicity profile. Conclusions: H-Zt/g4-MMAE is superior in eradication of pancreatic cancer xenografts with favorable pharmacokinetic profiles and manageable toxicological activities. These findings warrant the transition of H-Zt/g4-MMAE into clinical trials in the future. [ABSTRACT FROM AUTHOR]

Plant JASMONATE ZIM-DOMAIN (JAZ) genes play crucial roles in regulating the biosynthesis of specialized metabolites and stressful responses. However, understanding of JAZs controlling these biological processes lags due to numerous JAZ copies. Here, we found that two leaf-specific CwJAZ4/9 genes from Curcuma wenyujin are strongly induced by methyl-jasmonate (MeJA) and negatively correlated with terpenoid biosynthesis. Yeast two-hybrid, luciferase complementation imaging and in vitro pull-down assays confirmed that CwJAZ4/9 proteins interact with CwMYC2 to form the CwJAZ4/9-CwMYC2 regulatory cascade. Furthermore, transgenic hairy roots showed that CwJAZ4/9 acts as repressors of MeJA-induced terpenoid biosynthesis by inhibiting the terpenoid pathway and jasmonate response, thus reducing terpenoid accumulation. In addition, we revealed that CwJAZ4/9 decreases salt sensitivity and sustains the growth of hairy roots under salt stress by suppressing the salt-mediated jasmonate responses. Transcriptome analysis for MeJA-mediated transgenic hairy root lines further confirmed that CwJAZ4/9 negatively regulates the terpenoid pathway genes and massively alters the expression of genes related to salt stress signaling and responses, and crosstalks of multiple phytohormones. Altogether, our results establish a genetic framework to understand how CwJAZ4/9 inhibits terpenoid biosynthesis and confers salt tolerance, which provides a potential strategy for producing high-value pharmaceutical terpenoids and improving resistant C. wenyujin varieties by a genetic approach., (© 2024 John Wiley & Sons Ltd.)

The oncogenic role of circRNA in cancers including esophageal cancer (EC) has been well studied. However, whether and how circRNAs are involved in cancer cell metabolic processes remains largely unknown. Here, we reported that circRNA, circHIPK3, is highly expressed in ESCC cell lines and tissues. Knockdown of circHIPK3 significantly restrained cell proliferation, colony formation, migration, and invasion in vitro and inhibited tumor growth in vivo. Mechanistically, circHIPK3 was found to act as a ceRNA by sponging miR-637 to regulate FASN expression and fatty acid metabolism in ESCC cells. Anti-sense oligonucleotide (ASO) targeting circHIPK3 substantially inhibited ESCC both in vitro and in vivo. Therefore, these results uncover a modulatory axis constituting of circHIPK3/miR-637/FASN may be a potential biomarker and therapeutic target for ESCC in the clinic., (© 2024. The Author(s).)

Objective: To explore the mediating effect of coping style between illness perception and fear of cancer recurrence in patients undergoing radical prostatectomy., Methods: A questionnaire survey was carried out in 254 eligible patients who underwent radical prostatectomy in the urology department of two comprehensive tertiary hospitals in Wenzhou City from June 2022 to December 2022. The questionnaires include the general data questionnaire, brief illness perception questionnaire (BIPQ), Medical Coping Modes Questionnaire (MCMQ) and Fear of Progression Questionnaire-Short Form (FoP-Q-SF). A structural equation model was used to analyze the mediating effect of coping style between illness perception and fear of cancer recurrence., Results: The score of fear of cancer recurrence in prostate cancer patients is (30.08 ± 10.11). Illness perception, avoidance, and surrender coping styles could forward prediction fear of cancer recurrence (P=0.001, P=0.019, P=0.001); facing coping styles can negatively predict fear of cancer recurrence (P=0.001). Coping style played a part of the mediating role between illness perception and fear of cancer recurrence, and the mediating effect is 0.150，which accounted for 47.62% of the total effect., Conclusion: Coping style is a mediator between illness perception and fear of cancer recurrence in patients undergoing radical prostatectomy. Doctors and nurses should reduce patients' negative perception, guide them to adopt positive coping strategies, and thereby reduce their fear of cancer recurrence.

Obstructive azoospermia is a kind of common clinical disease, which often happens in epididymis, vas deferens and ejaculatory duct obstruction and accounts for male infertility. Azoospermia, which is caused by obstruction of the ejaculatory duct, is relatively rare. but the development of endoscopic technology now allows the disease to be resolved by surgery. In the past, transurethral resection of ejaculatory duct (TURED) has been used to treat patients with ejaculatory duct obstruction and azoosperm-free disease. But with the further study of TURED operation in recent years, as well as in patients with postoperative follow-up results showed that the urine - tube reflux and retrograde ejaculation ejaculation the incidence of iatrogenic injury is higherduring the TURED.therefore, in recent years, some experts committed to seeking a less damage and better effect of the surgical method. With the continuous development of endoscopic technology, seminal vesiculoscopy has come into being, which also creates conditions for further understanding the structure and function of the seminal vesicle and distal seminal duct, accurate diagnosis and minimally invasive treatment of the currently discovered diseases. In this article, we summarized the semen abnormalities associated with seminal vesiculoscopy in the treatment of ejaculatory duct obstruction, and also reviewed the surgical methods, clinical application and research progress of seminal vesiculoscopy.

Wenzhou has improved its environmental quality because of comprehensive environmental remediation; nevertheless, the semen quality of infertile males remains unclear. This study determined whether better environmental quality improved semen quality in this region. We recorded semen quality data from 22 962 infertile males from January 2014 to November 2019 at the Center for Reproductive Health of The First Affiliated Hospital of Wenzhou Medical University (Wenzhou, China). Patients were predominantly 30-35 years old (33.1%) and workers (82.0%), with high school education or lower (77.6%); more than a half of the patients (52.6%) were Wenzhou household registration; and most patients (77.5%) had abnormal semen quality. Patients who were older than 40 years and workers, and those with Wenzhou household registration, had significantly worse semen quality (all P < 0.05). From 2014 to 2019, progressive sperm motility, total sperm motility, and semen volume showed increasing linear trends in all patients (P = 0.021, 0.030, and 0.005, respectively), yet normal sperm morphology showed a linearly decreasing trend (P = 0.046). Sensitivity analyses for subgroups yielded similar results. In conclusion, the improvement of environmental quality and better function of the accessory glands are associated with progressive sperm motility, total sperm motility, and semen volume. Normal sperm morphology is influenced by occupational exposures and personal lifestyle and does not improve with environmental quality., Competing Interests: None

Plant-derived monoterpene indole alkaloids (MIAs) from Uncaria rhynchophylla (UR) have huge medicinal properties in treating Alzheimer's disease, Parkinson's disease, and depression. Although many bioactive UR-MIA products have been isolated as drugs, their biosynthetic pathway remains largely unexplored. In this study, untargeted metabolome identified 79 MIA features in UR tissues (leaf, branch stem, hook stem, and stem), of which 30 MIAs were differentially accumulated among different tissues. Short time series expression analysis captured 58 pathway genes and 12 hub regulators responsible for UR-MIA biosynthesis and regulation, which were strong links with main UR-MIA features. Coexpression networks further pointed to two strictosidine synthases (UrSTR1/5) that were coregulated with multiple MIA-related genes and highly correlated with UR-MIA features (r > 0.7, P < 0.005). Both UrSTR1/5 catalyzed the formation of strictosidine with tryptamine and secologanin as substrates, highlighting the importance of key residues (UrSTR1: Glu309, Tyr155; UrSTR5: Glu295, Tyr141). Further, overexpression of UrSTR1/5 in UR hairy roots constitutively increased the biosynthesis of bioactive UR-MIAs (rhynchophylline, isorhynchophylline, corynoxeine, etc), whereas RNAi of UrSTR1/5 significantly decreased UR-MIA biosynthesis. Collectively, our work not only provides candidates for reconstituting the biosynthesis of bioactive UR-MIAs in heterologous hosts but also highlights a powerful strategy for mining natural product biosynthesis in medicinal plants., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The coronavirus disease 2019 (COVID-19) pandemic caused extensive loss of life worldwide. Further, the COVID-19 and influenza mix-infection had caused great distress to the diagnosis of the disease. To control illness progression and limit viral spread within the population, a real-time reverse-transcription PCR (RT-PCR) assay for early diagnosis of COVID-19 was developed, but detection was time-consuming (4-6 h). To improve the diagnosis of COVID-19 and influenza, we herein developed a recombinase polymerase amplification (RPA) method for simple and rapid amplification of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the causative agent of COVID-19 and Influenza A (H1N1, H3N2) and B (influenza B). Genes encoding the matrix protein (M) for H1N1, and the hemagglutinin (HA) for H3N2, and the polymerase A (PA) for Influenza B, and the nucleocapsid protein (N), the RNA-dependent-RNA polymerase (RdRP) in the open reading frame 1ab (ORF1ab) region, and the envelope protein (E) for SARS-CoV-2 were selected, and specific primers were designed. We validated our method using SARS-CoV-2, H1N1, H3N2 and influenza B plasmid standards and RNA samples extracted from COVID-19 and Influenza A/B (RT-PCR-verified) positive patients. The method could detect SARS-CoV-2 plasmid standard DNA quantitatively between 10 2 and 10 5 copies/ml with a log linearity of 0.99 in 22 min. And this method also be very effective in simultaneous detection of H1N1, H3N2 and influenza B. Clinical validation of 100 cases revealed a sensitivity of 100% for differentiating COVID-19 patients from healthy controls when the specificity was set at 90%. These results demonstrate that this nucleic acid testing method is advantageous compared with traditional PCR and other isothermal nucleic acid amplification methods in terms of time and portability. This method could potentially be used for detection of SARS-CoV-2, H1N1, H3N2 and influenza B, and adapted for point-of-care (POC) detection of a broad range of infectious pathogens in resource-limited settings., (© 2022 The Authors. Journal of Medical Virology published by Wiley Periodicals LLC.)

Dendrobium officinale is an important traditional Chinese medicinal herb, and the stem tissue is the main medicinal that is harvested from D. officinale. Recently, the first viroid was identified from D. officinale in China, and it has been named Dendrobium viroid (DVd). Whether DVd interferes with metabolic pathways in dendrobium plants and affects the medicinal value of the host is unknown. In this study, metabolomics data from stem tissues supported by transcriptome studies were used to investigate how metabolism modulate of D. officinale is altered by DVd infection. Our results show that metabolism of D. officinale is reprogrammed in many ways during DVd infection, and this is reflected by significant changes in the levels of flavonoids, alkaloids, and phenolic acids. Furthermore, we found that DVd infection significantly decreased the accumulation of flavonoids and alkaloid metabolites in infected stems, and the decreases in these metabolites appears to affect the medicinal components of the infected plants, weakening the host antiviral immune response as well. Conversely, phenolic acids occupy a larger proportion of the up-regulated metabolites from DVd infection in comparison with the mock-inoculated control, and the increase in the total phenolic acids may reflect the activation of the pathogen defense response in D. officinale. Taken together, our results provide an interesting overview and give a better understanding of the relationship between metabolism and DVd infection in the orchid D. officinale., (Copyright © 2021 Elsevier B.V. All rights reserved.)

With the increasing awareness of men's health, more and more clinical studies are reported on the relationship of testicular microlithiasis (TM) with male infertility. TM is a relatively rare disease characterized by multiple microcalcifications (<3 mm) in the seminiferous tubules of the testis. This review summarizes the findings in the studies of the diagnosis, epidemiology, correlation with male fertility, treatment, and follow-up of TM in recent years. Many researches show that TM has a certain correlation with the decline of male fertility, which, however, is not identically agreed on at home and abroad. As for the treatment of TM complicated with male infertility, there is still a lack of effective and reliable drugs and strategies and a need for many basic and clinical studies.

This study analyzed the trend in semen quality of infertile male patients in Wenzhou, China, based on the data obtained from 38 905 patients during 2008-2016 in The First Affiliated Hospital of Wenzhou Medical University (Wenzhou, China). The results showed that only 24.9% of the patients had normal semen quality. For the semen quality of infertile male patients, that of the workers and 40-year-olds was significantly worse than the other occupational and age groups. For all the infertile patients, low semen volume, asthenozoospermia, and teratozoospermia accounted for 8.4%, 50.5%, and 54.1%, respectively. During 2008-2016, the annual mean percentage of fast forward motile spermatozoa, percentage of total forward motile spermatozoa, and percentage of spermatozoa with normal morphology decreased linearly with slopes of -2.11, -2.59, and -0.70, respectively. The proportion of patients with asthenozoospermia and multi-abnormal spermatozoa increased during 2008-2016 with slopes of 4.70 and 4.87, respectively, while for low semen volume, it decreased with a slope of -0.47 in the same time period. The proportion of patients with teratozoospermia increased from 2008 to 2011 and from 2011 to 2016 with slopes of 17.10 and 2.09, respectively. In general, the deteriorating trend of semen quality of infertile male patients in Wenzhou was obvious. Future efforts should be made to reveal the adverse influences on semen quality, such as occupational exposure, environmental quality, and living habits. Furthermore, more pervasive reproduction health education is necessary., Competing Interests: None

Purpose: to describe the methodology of transurethral seminal vesiculoscopy and the anatomy of the area of the verumontanum, and to determine the safety of this procedure, especially in terms of postoperative complications., Methods: This retrospective observational study enrolled 144 patients with intractable hematospermia from May 2011 and August 2019. A 4.5/6.5-Fr vesiculoscope was inserted into the seminal vesicle to deal with the positive findings. The solution of quinolones was used to rinse each seminal vesicle., Results: In this study, Transurethral seminal vesiculoscopy was successfully performed in 139 patients (96.53%). Hematospermia was alleviated or disappeared in 116 (80.56%) patients by less than half a year after surgery. Common intraoperative manifestations were hemorrhage, stones, utricle polyps and cysts. The surgical approach in our study were categorized into four types, including 24 (16.7%), 73 (50.7%), 42 (29.2%), and 5 (3.5%) cases in Type A (natural opening of the ejaculatory duct), B (trans-duct fenestration), C (trans-utricle fenestration), and D (not founded), respectively. Sexual function change was recorded in 12 patients of 111 patients, all by the method of trans-utricle fenestration, including 8 (7.21%), 3 (2.70%), and 1 (0.90%) patients in shorter intravaginal ejaculatory latency time, worse erection hardness and loss of orgasm, respectively., Conclusion: Transurethral seminal vesiculoscopy is an effective and safe procedure for the management of hematospermia. The anatomy of the distal seminal tract should be understood more deeply and Wu'method (uncover-curtain method) needs to be promoted to verify its universality and safety. Besides, the complications of the function dysfunction should be discussed in the future in multi-center clinical trials.

Background and Aims: There is a lack of research on metastatic renal pelvis cell carcinoma in the current literature. In this study, we aimed to detect distant metastatic patterns in renal pelvis cell carcinoma, and illustrated the affection of different metastatic sites, surgery to primary site and chemotherapy on prognosis outcomes in patients with diverse conditions., Methods: We collected data between 2010 and 2015 from the Surveillance, Epidemiology and End Results database. Kaplan-Meier analysis with log-rank test was used for survival comparisons. Multivariate Cox regression model was employed to analyze the effect of distant metastatic sites on overall survival (OS) and cancer-specific survival (CSS)., Results: A total of 424 patients were included in the analysis, the median follow-up time was 5 months (interquartile range (IQR): 2-12) and 391 deaths (92.2%) in all patients were recorded. Among them, 192 (45.3%), 153 (36.1%), 137 (32.3%) and 127 (30.0%) patients were diagnosed with lung, bone, liver and brain metastases, respectively, while only 12 (2.8%) patients had brain metastases. The bi-organ, tri-organ and tetra-organ metastatic pattern was found in 135 (31.8%), 32 (7.5%) and 11 (2.6%) patients, respectively. The multivariate Cox analyses showed that distant lymph nodes (DL) metastases was not an independent prognostic factor for both OS and CSS (OS: Hazard ratios (HR) = 1.1, 95% CI = 0.8-1.4, P = 0.622; CSS: HR = 1.0, 95% CI = 0.8-1.3, P = 0.906). Besides, there was no significant difference of survival in patients with T3-T4 stage (OS: HR = 0.8, 95% CI = 0.5-1.2, P = 0.296; CSS: HR = 0.8, 95% CI = 0.5-1.2, P = 0.224), N2-3 stage (OS: HR = 0.8, 95% CI = 0.5-1.3, P = 0.351; CSS: HR = 0.7, 95% CI = 0.4-1.2, P = 0.259) and multi-organ metastases (OS: HR = 0.8, 95% CI = 0.5-1.3, P = 0.359; CSS: HR = 0.7, 95% CI = 0.4-1.2, P = 0.179) between surgery to primary site group and no-surgery to primary site group., Conclusion: we described the metastatic patterns of mRPCC and the prognosis outcomes of DL metastases, surgery to primary site and chemotherapy. Our findings provide more information for clinical therapeutic intervention and translational study designs.

Background: Controversial effectiveness of chemotherapy and still poor prognosis prompt us to find better treatment options. This study targeted at investigating whether adding radiotherapy based on chemotherapy can effectively improve the prognosis of patients, especially for advanced penile cancer., Methods: Data were obtained from the Surveillance, Epidemiology, and End Results database (SEER*Stat software V.8.3.5; USA; Accession numbers: 13693-Nov2015 and lh8N79l2), and the survival curves were conducted using the Kaplan-Meier method. Univariate and multivariate cox regression models were performed in order to determine the hazard ratios (HRs) with 95% confidence intervals (CIs) for penile cancer-specific survival (PCSS). Subgroup analysis via multivariate Cox models were conducted to discovery the different effect in population with different features., Results: The median follow-up time was 25 months, the 2-year PCSS was 52.98% in the chemoradiotherapy group and 55.81% in the chemotherapy group. In multivariate analysis of all patients, combined chemoradiotherapy was not associated with PCSS (HR =0.90, 95% CI: 0.63-1.29, P=0.572). In subgroup analysis, chemoradiotherapy improved the PCSS in N3 patients compared to these patients without therapy of radiotherapy (HR =0.54, 95% CI: 0.30-0.98, P=0.043)., Conclusions: Our study demonstrated a significant correlation of chemoradiotherapy with improved cancer-specific survival of penile cancer (PeCa) in N3 patients. Prospective international multicenter studies are necessary in order to improve prognosis for patients with advanced penile cancer., Competing Interests: Conflicts of Interest: Both authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/tau-20-1044). The authors have no conflicts of interest to declare., (2020 Translational Andrology and Urology. All rights reserved.)

Background: Programmed death ligand 1 (PD-L1) immunotherapy remains poorly efficacious in colorectal cancer (CRC). The recepteur d'origine nantais (RON) receptor tyrosine kinase plays an important role in regulating tumor immunity., Aim: To identify the patterns of RON and PD-L1 expression and explore their clinical significance in CRC., Methods: Gene expression data from the Gene Expression Omnibus database (GEO; n = 290) and patients at the First Affiliated Hospital, Zhejiang University School of Medicine (FAHZUSM; n = 381) were analyzed to determine the prognostic value of RON and PD-L1 expression within the tumor microenvironment of CRC. HT29 cell line was treated with BMS-777607 to explore the relationship between RON activity and PD-L1 expression. Signaling pathways and protein expression perturbed by RON inhibition were evaluated by cellular immunofluorescence and Western blot., Results: In the GEO patient cohort, cut-off values for RON and PD-L1 expression were determined to be 7.70 and 4.3, respectively. Stratification of patients based on these cutoffs demonstrated that high expression of RON and PD-L1 was associated with a poor prognosis. In the FAHZUSM cohort, rates of high expression of RON in tumor cells, high PD-L1 expression in tumor cells and tumor infiltrating monocytes, and both high RON and high PD-L1 expression in the tumor microenvironment were 121 (32%), 43 (11%), 91 (24%), and 51 (13.4%), respectively. High expression of RON was significantly correlated with high expression of PD-L1 in the tumor cell compartment ( P < 0.001). High expression of RON and that of PD-L1 were independent prognostic factors for poorer overall survival. Concurrent high expression of both RON and PD-L1 in the tumor microenvironment was significantly associated with a poor prognosis. In vitro , BMS-777607 inhibited the phosphorylation of RON, inhibited PD-L1 expression, and attenuated activation of the ERK1/2 and AKT signaling pathways in CRC cells., Conclusion: RON, PD-L1, and their crosstalk are significant in predicting the prognostic value of CRC. Moreover, phosphorylation of RON upregulates PD-L1 expression, which provides a novel approach to immunotherapy in CRC., Competing Interests: Conflict-of-interest statement: The authors confirm that there are no known conflicts of interest associated with this publication. The manuscript has been read and approved by all authors and there are no other persons who satisfied the criteria for authorship but are not listed. We further confirm that the order of authors listed in the manuscript has been approved by all of authors., (©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.)

With the continuous improvement of living conditions, increasing attention is being drawn to the genitourinary health of males, which has boomed the development of uroandrology in recent years. Refractory hemospermia, infertility, and perineal pain are commonly seen in some male patients, and mainly relied on medical imaging for diagnosis in the past, which, however, has a high pseudopositive rate and cannot give an etiological explanation. Patients with these diseases often adopt conservative treatments such as medication and physiotherapy, often with poor prognosis, and those suffering frequent recurrence used to be treated by transurethral resection, laparoscopic surgery or open surgery, which are now rarely employed due to their high rate of postoperative complications, slow recovery, and easy recurrence. In recent years, transurethral seminal vesiculoscopy has gained a wide application in the diagnosis and treatment of the above-mentioned uroandrological diseases and shown its advantages of high clinical effectiveness and low incidence of complications. The review updates on the indications, methods, skills and clinical application of transurethral seminal vesiculoscopy.

Background: To develop a nomogram for predicting cancer-specific survival (CSS) of patients with non-metastatic primary adenocarcinoma of the bladder (NMACB)., Methods: We used a retrospective cohort study design. Patient data were obtained from the SEER database, univariate and multivariate Cox regression analyses were performed to identify factors associated with CSS. A nomogram visualization model was established using R language software to predict survival rate. Harrell's concordance index (C-index), area under the receiver operating characteristic (ROC) curve (AUC) in addition to calibration plots were used to assess the performance of the model., Results: A total of 1,635 patients were included in the study. A multivariate Cox regression model indicated that age, histological type, grade, stage, and surgery were independent covariates associated with CSS. Using these prognostic factors, a nomogram was constructed. Harrell's C indices for CSS were 0.729 in the training cohort and 0.716 in the validation cohort. AUC values were 0.769, 0.735 and 0.724 for 1, 3, and 5-year in the training cohort, and 0.738, 0.727 and 0.713 for 1, 3 and 5-year in the validation cohort, respectively. The AUC values and calibration plots indicated that the nomogram provided good predictive performance., Conclusions: A nomogram for predicting CSS in patients with NMACB was developed to assist clinicians in the accurate prediction of mortality risk to allow them to recommend a personalized treatment modality., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/tcr-20-354). The authors have no conflicts of interest to declare., (2020 Translational Cancer Research. All rights reserved.)

Purpose: Triple-negative breast cancer (TNBC) is highly malignant and has poor prognosis and a high mortality rate. The lack of effective therapy has spurred our investigation of new targets for treating this malignant cancer. Here, we identified RON (macrophage-stimulating 1 receptor) and MET (MET proto-oncogene, receptor tyrosine kinase) as a prognostic biomarker and therapeutic targets for potential TNBC treatment., Materials and Methods: We analyzed RON and MET expression in 187 primary TNBC clinical samples with immunohistochemistry. We validated the targeted therapeutic effects of RON and MET in TNBC using three tyrosine kinase inhibitors (TKIs): BMS-777607, INCB28060, and tivantinib. The preclinical therapeutic efficacy of the TKIs was mainly estimated using a TNBC xenograft model., Results: Patients with TNBC had widespread, abnormal expression of RON and MET. There was RON overexpression, MET overexpression, and RON and MET co-overexpression in 63 (33.7%), 63 (33.7%), and 43 cases (23.0%), respectively, which had poor prognosis and short survival. In vivo, the TKI targeting RON ant MET inhibited the activation of the downstream signaling molecules, inhibited TNBC cell migration and proliferation, and increased TNBC cell apoptosis; in the xenograft model, they significantly inhibited tumor growth and shrank tumor volumes. The TKI targeting RON and Met, such as BMS-777607 and tivantinib, yielded stronger anti-tumor effects than INCB28060., Conclusion: RON and MET co-overexpression can be significant pathological characteristics in TNBC for poor prognosis. TKIs targeting RON and MET have stronger drug development potential for treating TNBC.

Background: To examine the association between age at diagnosis and cancer-specific mortality (CSM) in primary urachal adenocarcinoma., Methods: The data was obtained from the National Cancer Institute's Surveillance, Epidemiology, and End Results program (SEER). A total of 393 patients were included in the study. Smooth curve fitting and two-piecewise Cox proportional hazards models were used to identify the nonlinearity between the age at initial diagnosis and cancer-specific survival rate. Survival time between different groups was compared using Kaplan-Meier survival curves and the log-rank test., Results: Using smooth curve fitting we found that the relationship between age at diagnosis and cancer-specific survival takes on a U-shaped curve. The inflection point that we identified for the age at initial diagnosis was 60 years. The log-likelihood ratio test (P<0.05) indicated that the two-piecewise Cox regression model was more appropriate for fitting the correlation of age at diagnosis and CSM. The two-piecewise Cox regression model showed that when the age was <60 years, reduced risk of CSM was significantly associated with increased age (HR: 0.95, P=0.0002). Conversely, when age was >60 years, increased risk of CSM was significantly associated with increased age (HR: 1.05, P=0.0499)., Conclusions: In summary, our study suggested that the relationship between age at diagnosis and cancer-specific survival is nonlinear, and takes on a U-shaped curve. Both younger and older age at initial diagnosis age were associated with increased CSM., Competing Interests: Conflict of Interest: All authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/tau-19-863). The authors have no conflicts of interest to declare., (2020 Translational Andrology and Urology. All rights reserved.)

Background: To develop a nomogram to predict cancer-specific survival (CSS) in patients with metastatic testicular germ cell tumors (mTGCTs)., Methods: Data were obtained from the Surveillance, Epidemiology, and End Results database. Univariate and multivariate Cox regression models were used to identify factors associated with CSS. Survival times between different groups were compared using Kaplan-Meier survival curves and the log-rank test. A nomogram visualization model was established using the R language to predict survival rates. Harrell's concordance index (C-index), the area under the receiver operating characteristic curve (AUC) and calibration plots were used to assess the performance of the model., Results: We analyzed the data of 949 patients. The median follow-up time was 32 months (range 0 to 83 months), and 224 (23.60%) patients died before the last follow-up, of whom 193 (20.33%) died of mTGCTs. The site of distant metastases was an independent prognostic factor for CSS. Compared to patients without involvement of the corresponding organ, patients with bone, brain, liver, and lung involvement had worse CSS. We also found that age, histological type, surgery, radiation therapy, chemotherapy, metastatic site and insurance status affected the CSS of patients with mTGCTs. We used these prognostic factors to construct our nomogram. Harrell's C-index for CSS was 0.739. The AUC and calibration plots indicated good performance of the nomogram., Conclusions: A nomogram for predicting CSS in patients with mTGCTs has been developed, which can help patients and clinicians accurately predict mortality risk and recommend personalized treatment modalities., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/tcr.2020.03.59). The authors have no conflicts of interest to declare., (2020 Translational Cancer Research. All rights reserved.)

The novel coronavirus disease 2019 (COVID-19) broke out in December 2019 and has been rapidly escalating throughout the world. Clinical findings show that the patients with either symptomatic or asymptomatic COVID-19 can be a potential source of infection. Although respiratory droplets and close contact are considered to be the main routes of transmission, there is the possibility of aerosol transmission in a relatively closed environment. The nucleic acid of the novel coronavirus can be detected in nasopharyngeal swabs, sputum and other lower respiratory tract secretions, blood, feces, urine and so on, but whether it exists in the semen has not been confirmed. It is reported that the novel coronavirus may affect the testis that highly expresses angiotensin-converting enzyme 2 (ACE2) and theoretically the semen is a possible carrier of the virus considering the fact that it is discharged from the same channel as the urine. Andrology laboratorians are exposed to most of the specimens above, including semen, and some open operations in the laboratory increase the risk of aerosol generation. Therefore, corresponding protective procedures are necessitated in andrology laboratories to reduce the risk of infection during the outbreak of COVID-19. Based on the knowledge and experience available as regards the pandemic and the characteristics of the work in the andrology laboratory, we summarize some biosafety points for andrology laboratorians to attend to during the outbreak of COVID-19.

RON (recepteur d'origine nantais) and MET (hepatocyte growth factor receptor) are tyrosine kinase receptors. Various cancers have aberrant RON and MET expression and activation, which contribute to cancer cell proliferation, invasiveness, and metastasis. Here, we explored RON and MET expression in pancreatic cancer and their relationship with overall survival (OS) time, and evaluated their significance as therapeutic targets of tyrosine kinase inhibitors in pancreatic cancer. We enrolled 227 patients with pancreatic cancer in the study. RON and MET expression was analyzed by immunohistochemical staining. Four human pancreatic cancer cell lines expressing variable levels of RON or MET and four MET superfamily inhibitors (BMS777607, PHA665752, INCB28060, Tivantinib) were used. The effect of the four tyrosine kinase inhibitors on cell viability, migration, and apoptosis were determined using cell viability, scratch wound healing, and Caspase-Glo 3/7 assays. Cellular signaling was analyzed by immunoprecipitation and western blotting. The therapeutic efficacy of the tyrosine kinase inhibitors was determined with mouse xenograft pancreatic cancer models in vivo . There was wide aberrant RON and MET expression in the cancer tissues. In 227 pancreatic cancer samples, 33% had RON overexpression, 41% had MET overexpression, and 15.4% had RON and MET co-overexpression. RON and MET expression were highly correlated. RON and MET expression levels were significantly related to OS. Patients with RON and MET co-overexpression had poorer OS. BMS777607 and PHA665752 inhibited pancreatic cancer cell viability and migration, and promoted apoptosis by inhibiting RON and MET phosphorylation and further inhibiting the downstream signaling pathways in vitro . They also inhibited tumor growth and further inhibited phosphorylated (phosphor)-RON and phospho-MET expression in the mouse xenograft models in vivo effectively. INCB28060, which inhibits the MET signaling pathway alone, was not effective. RON and MET can be important indicators of prognosis in pancreatic cancer. Tyrosine kinase inhibitors targeting RON and MET in pancreatic cancer are a novel and potential approach for pancreatic cancer therapy., (Copyright © 2019 Hu, Xu, Hong, Wu, Qian, Weng, Liu, Tang, Wang and Yao.)

Aims: Alzheimer's disease (AD) is closely related to abnormal glucose metabolism in the central nervous system. Progesterone has been shown to have obvious neuroprotective effects in the pathogenesis of AD, but the specific mechanism has not been fully elucidated. Therefore, the purpose of this study was to investigate the effect of progesterone on the glucose metabolism of neurons in amyloid precursor protein (APP)/presenilin 1 (PS1) mice and Aβ-induced AD cell model., Materials and Methods: APP/PS1 mice were treated with 40 mg/kg progesterone for 40 days and primary cultured cortical neurons were treated with 1 μM progesterone for 48 h.Then behavior tests,2-NBDG glucose uptake tests and the protein levels of glucose transporter 3 (GLUT3), GLUT4, cAMP-response element binding protein (CREB) and proliferator-activated receptor γ (PPARγ) were examined., Key Findings: Progesterone increased the expression levels of GLUT3 and GLUT4 in the cortex of APP/PS1 mice, accompanied by an improvement in learning and memory. Progesterone increased the levels of CREB and PPARγ in the cerebral cortex of APP/PS1 mice. In vitro, progesterone increased glucose uptake in primary cultured cortical neurons, this effect was blocked by the progesterone receptor membrane component 1 (PGRMC1)-specific blocker AG205 but not by the progesterone receptor (PR)-specific blocker RU486. Meanwhile, progesterone increased the expression of GLUT3, GLUT4, CREB and PPARγ, and AG205 blocked this effect., Significance: These results confirm that progesterone significantly improves the glucose metabolism of neurons.One of the mechanisms of this effect is that progesterone upregulates protein expression of GLUT3 and GLUT4 through pathways PGRMC1/CREB/GLUT3 and PGRMC1/PPARγ/GLUT4., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Tetraena mongolica Maxim, a relict originating from the Tertiary Period, is an endemic species of Zygophyllaceae in China. Three new monoterpenoids ( 1 - 3 ), two new phenols ( 4 , 5 ) with unusual O -sulfoglucosyl groups, a new flavonoid ( 6 ), and nine known compounds were isolated from the leaves of T. mongolica . The structures of these compounds were determined by interpretation of NMR, MS, and ECD data. Some of the isolated compounds showed protective effects on HEK 293t cells damaged by CdCl 2 , with IC 50 values being 55.7 and 80.3 μM for compounds 7 and 8 , respectively, at the time point of 48 h after treatment.

Objective: To investigate the status quo of the diagnosis and treatment of male urethritis (MU) in urology and andrology., Methods: According to The Guidelines for Clinical Diagnosis and Treatment of Sexually Transmitted Diseases (2017), we designed 27 questions on the prevalence, diagnosis, treatment, and prognosis of MU. Using these questions, we conducted a questionnaire investigation among urological, andrological and other relevant clinicians with different professional titles, followed by an analysis of the compliance of the doctors to the Guidelines., Results: Totally, 116 valid questionnaires were collected from 86 urological, 28 andrological and 2 other relevant doctors, including 22 professors, 36 associate professors, 40 attending doctors and 16 resident doctors. MU was found mostly in those aged 20－40 years and more than half of the patients had a history of unclean sex, gonococcal urethritis significantly less prevalent than non-gonococcal, with Ureaplasma urealyticum as the most common pathogen of non-gonococcal urethritis. As for the compliance to the Guidelines in the diagnosis of MU, 22.73% of the professors, 16.67% of the associate professors, 15.00% of the attending doctors and 12.50% of the resident doctors examined the eyes, mouth and perianus (P > 0.05), 40.91% of the professors, 58.33% of the associate professors, 40.00% of the attending doctors and 37.50% of the resident doctors conducted HIV and syphilis screening (P > 0.05), and 86.36% of the professors, 77.78% of the associate professors, 70.00% of the attending doctors and 75.00% of the resident doctors performed genital mycoplasma screening (P > 0.05). Concerning the treatment of MU, 50.00% of the professors, 47.22% of the associate professors, 22.50% of the attending doctors and 43.75% of the resident doctors used anti-Chlamydia trachomatis drugs for gonococcal urethritis (P > 0.05), 0.00% of the professors, 11.11% of the associate professors, 5.00% of the attending doctors and 31.25% of the resident doctors prescribed 1g single-dose oral azithromycin for non-gonococcal urethritis (P < 0.05), 13.64% of the professors, 33.33% of the associate professors, 17.50% of the attending doctors and 6.25% of the resident doctors medicated persistent or recurrent non-gonococcal urethritis for >4 weeks (P > 0.05), 63.64% of the professors, 83.33% of the associate professors, 57.50% of the attending doctors and 62.50% of the resident doctors treated asymptomatic trachomatis and mycoplasma infections according to the proposed medication in the Guidelines (P > 0.05). As regards the results of treatment, the cure rate of gonococcal urethritis was 100.00% by professors, 97.22% by associate professors, 95.00% by attending doctors and 81.25% by resident doctors (P > 0.05), and that of non-gonococcal urethritis was 86.36% by professors, 61.11% by associate professors, 62.50% by attending doctors and 37.50% by resident doctors (P < 0.05)., Conclusions: Urological and andrological clinicians do not strictly follow the Guidelines in the diagnosis and treatment of male urethritis. There are significant differences in the dosing of azithromycin and results of treatment of non-gonococcal urethritis among doctors with different professional titles, but not in the other aspects.

Objective: To investigate the effects of low-dose PDE5 inhibitors on metabolic parameters and erectile function in ED patients with subclinical metabolic syndrome (SCMS)., Methods: Totally, 132 ED patients, aged 21－61 (mean 34.5) years, were treated in the Andrology Clinic of the First Hospital of Wenzhou Medical University from April 2017 to May 2018. According to the diagnostic criteria, we divided the patients into groups A (simple ED, n = 40), B (ED with SCMS, n = 34) and C (ED with MS, n = 58) to receive 3 months of oral administration of tadalafil at 5 mg qd at bedtime, and followed them up for 3 months after drug withdrawal. During the treatment, we advised the patients to keep a healthy diet, change bad habits, participate in regular physical exercise, and maintain psychological balance. Before and right after medication and at 3 months after drug withdrawal, we recorded the changes in the IIEF-5 scores, abdominal circumference, blood pressure and levels of fasting blood sugar (FBS), triglyceride (TG) and high-density lipoprotein (HDL) of the patients., Results: The IIEF-5 scores showed statistically significant differences at different time points between groups A and C (P < 0.01), remarkably higher right after treatment than before treatment and at 3 months after drug withdrawal in group B (19.71 ± 2.40 vs 10.21 ± 3.92 and 16.29 ± 2.41, P < 0.01). At 3 months after drug withdrawal, the abdominal circumference was significantly smaller in group A than in B and C (［78.10 ± 6.00］ vs ［84.15 ± 8.17］ and ［91.53 ± 11.49］ cm, P < 0.01) and the HDL level lower in group C than in A and B (［0.96 ± 0.15］ vs ［1.27 ± 0.14］ and ［1.16 ± 0.2］] mmol/L, P < 0.01). Systolic blood pressure exhibited statistically significant differences between any two time points in group C (P < 0.05 or P < 0.01) but not in group A (P > 0.05) or B (P > 0.05). Diastolic blood pressure was markedly lower in group B right after medication and at 3 months after drug withdrawal than before treatment (［75.62 ± 10.70］ and ［74.65 ± 9.90］ vs ［78.00 ± 11.42］ mmHg, P < 0.05), and so was it in group C (［82.19 ± 10.36］ and ［82.40 ± 10.09］ vs ［86.71 ± 12.32］ mmHg, P < 0.05), but manifested no significant difference between any two time points in group A (P > 0.05). There were statistically significant differences in the FBS level among different time points in groups A and C (P < 0.05) but not in B between post-treatment and 3 months after drug withdrawal (［5.34 ± 0.60］ vs ［5.36 ± 0.40］ mmol/L, P > 0.05), and so were there in the TG level among different time points in groups A and C (P < 0.05) but not in B between pre- and post-treatment (［1.80 ± 0.98］ vs ［1.64 ± 1.19］ mmol/L, P > 0.05)., Conclusions: Periodic administration of low-dose sustained-release PDE5 inhibitors with health education and lifestyle guidance may reverse ED with SCMS and improve most of the related metabolic parameters.

The nuclear factor E2-related factor 2 (NRF2)-Kelch-like ECH-associated protein 1 (KEAP1) signaling cascades is a key transcriptional pathway governing cellular oxidative stress and tumor development. Mammalian hepatitis B X-interacting protein (HBXIP) has critical roles in modulating cancer malignance and tumor progression. However, whether HBXIP interacts with KEAP1 and NRF2 is unclear. Here, we found that HBXIP can effectually compete with NRF2 for binding with KEAP1 protein via its highly conserved GLNLG motif. The HBXIP-mediated reduction in NRF2-KEAP1 complexes promotes NRF2 accumulation and nuclear entry, which facilities the activation of antioxidant response element (ARE)-dependent signaling cascades, thereby reducing the accumulation of endogenous cellular reactive oxygen species (ROS). We also found a strong positive correlation between HBXIP expression and NRF2 expression in breast cancer cells, tissue microarrays and clinical breast cancer tissues. Furthermore, this positive correlation was further confirmed via analysis of 1905 clinical cases of breast carcinoma provided by the cancer genomics database cBioPortal. Strikingly, disrupting the HBXIP-KEAP1 axis via mutating the GLNLG motif of HBXIP leads to potent inhibition of the malignancy of breast carcinoma both in vivo and in vitro. Our findings broaden our understanding of HBXIP as a modulation factor of cellular oxidative stress and address a novel regulatory mechanism governing redox homeostasis and the progression of breast carcinoma.

Objective: To investigate the effect of the frenulum identification positioning method with a disposable suture device in circumcision for the prevention of postoperative penile frenulum malposition., Methods: Totally 212 patients with phimosis or redundant prepuce underwent circumcision from March 2015 to September 2016, including 109 cases of conventional circumcision (the control group) and 103 cases treated by frenulum identification positioning with a disposable suture device (the observation group). We observed the postoperative position of the penile frenulum and median raphe and compared the deviation angles of the frenulum between the two groups of patients., Results: The median of penile frenulum deviation angle (interquartile range) was 0 (3.56) in the observation group, significantly smaller than 12.41 (19.59) in the control (P <0.001, P = 0.000). And the rate of frenulum deviation was remarkably lower in the former (8.74% ［9/103］) than in the latter group (66.06% ［72/109］) (P <0.01)., Conclusions: Circumcision using the frenulum identification positioning method with a disposable suture device can effectively avoid postoperative penile frenulum malposition. With the advantages of safety and easy operation, it deserves clinical application and popularization.

Apoptosis plays an important role in intervertebral disc degeneration (IDD). Overwhelming evidence indicates that RASSF7 is essential for cell growth and apoptosis. Recently, it has been noted that the JNK signaling can be negatively regulated by suppressing phosphorylated-MKK7 activation during pro-apoptosis. We aimed to investigate the RASSF7 expression level in human degenerative nucleus pulposus (NP) cells and non-degenerative NP cells and the link between RASSF7-JNK with NP cells apoptosis. We harvested NP tissues from 20 IDD patients as disease group and 8 cadaveric donors as normal controls. We detected RASSF7 expression by Real-time-PCR and western blotting. Consequently, we found that the expression of RASSF7 was higher in non-degenerative group than in degenerative group (P<0.05). Overexpression of RASSF7 in degenerative NP cells led to decreased apoptosis rate than that in scramble group (P<0.05). Collectively, our findings suggest that RASSF7 plays an important role in human IDD and RASSF7 might be potentially developed as a curative agent.

Objective: To evaluate the clinical efficacy and safety of phosphodiesterase 5 (PDE-5) inhibitors for erectile dysfunction (ED) in patients with diabetes mellitus and provide some evidence for the clinical treatment of the disease., Methods: We searched MedMed, EMbase, Cochrane Library, CNKI, Wan Fang Data, VIP and ZADL for randomized controlled trials on PDE-5 inhibitors for ED in diabetic men and evaluated the methodology of the included trials with the Jadad scale. We used the erectile function domain in the IIEF (IIEF-EF), IIEF questions (IIEF-Q) 3 and 4, SEP-2 and -3, and Global Assessment Questions (GAQ) as the main evaluation indexes and employed the Review Manager 5. 1. 0 software for meta analysis., Results: A total of 13 studies were included, which were all high quality trials with Jadad score > 3. The IIEF-EF scores in 10 of the included studies were subjected to meta analysis using the random-effect model (REM), with a weighted mean difference (WMD) of 5.64 (95% CI 4.41 - 6.83, P < 0.001). The fixed-effect model (FEM) analysis of the IIEF-Q scores in 6 of the studies showed the WMD to be 0.96 (95% CI 0.83 -1.08, P < 0.001) for IIEF-Q3 and 1.11 (95% CI 0.98 - 1.25, P < 0.001) for IIEF-Q4. FEM analysis of the SEP-2 scores showed WMD = 17.67 (95% CI 12. 38 - 22. 97, P < 0.001) in 2 of the studies, and that of the SEP-3 scores WMD = 23.64 (95% CI 17. 49 - 29.79, P < 0.001) in 5 of the studies. The GAQ scores in 11 of the studies were subjected to REM analysis, with OR = 6. 20 and 95% CI 3.65 - 10.52 (P < 0.001). REM analysis was performed on the adverse reactions in 11 of the studies, with OR = 7.43 and 95% CI 4.11 - 13.44 (P < 0.001)., Conclusion: PDE-5 inhibitors can effectively and safely improve erectile function in patients with diabetes mellitus.

To breed a new yam cultivar of Dioscorea alata, the different and excellent germplasm resources were investigated within artificially cultivated population and some superior individuals, with a higher yield and medicinal properties, were selected. Considering results of the yield and medicinal properties during 2006-2013 cropping season, strains and lines were established and selected. As a result, the yield of the new developed cultivar (Wenshanyao No. 1, WSY01-1) reached 2217. 0 kg per 667 m2 (fresh weight) and 348.3 kg per 667 m2 (dry weight), and increased 23.8% and 23.9% comparing with control cultivars (landraces). Comparing with control cultivars, the level of polysaccharide, allantoin, and dioscin increased 36.9%, 48.3%, 20.9%, and reached 12.2%, 1.30%, 579.7 µg · g(-1), respectively. This result showed that the systematic selection method can significantly improve yield and medicinal properties of D. alata, and the developed " Wenshanyao No. 1" exhibits wide spreading prospects.

Objective: To determine the effects of Jiji decoction (Traditional Chinese Medicine) on the cognitive function and oxidative stress in mice with vascular dementia (VD) induced by cerebral ischemia/reperfusion., Methods: Thirty-two mice were randomly divided into nonnal group (n = 8), sham group (operation, but no cerebral ischemia/reperfusi6n, n = 8), model group (vascular dementia model induced by cerebral ischemia/reperfusion, n = 8), and Jiji decoction-treated group (vascular dementia model plus treatment with Jiji decoction, n = 8). Fourteen days of treatment after operation, the cognitive behavior was measured in step-through test, spatial probe test and platform test. Afterwards, to assess the levels of oxidative stress, the activity of superoxide dismutase(SOD) and content of malonaldehyde (MDA) in brain of these mice were measured., Results: Data from step-through test indicated that the escaping latency of Jiji decoction-treated group was prolonged and the error counts were decreased significantly ( P <0.01) compared with those of model group. Data from spatial probe test indicated that the time of entering darkroom, the time of climbing height and the time of entering bright room in Jiji decoction-treated group were shortened and the counts of climbing height were increased (P < 0.05-0.01) significantly compared with those of model group. Data from platform test showed that the escaping latency of Jiji decoction-treated group was prolonged significantly (P < 0.01) compared with that of model group. Compared with normal and sham group, the activity of SOD was decreased and the content of MDA was increased in model group significantly (P < 0.01). Compared with those of model group, the levels of SOD and MDA in Jiji decoction-treated group were improved significantly (P < 0.01)., Conclusion: Jiji decoction could improve cognitive function of VD mice. Its mechanism might be related with the inhibition of oxidative stiess in the brain.

Objective: To investigate the protective effect of phosphodiesterase type 5 inhibitors (tadalafil) on the testis following testicular ischemia-reperfusion injury in rats., Methods: Eighty-four healthy adult male SD rats were randomly and equally divided into groups A (sham operation), B (testicular torsion + low-dose tadalafil), C (testicular torsion + high-dose tadalafil), and D (testicular torsion + placebo). Models were established in the latter three groups by 7200 torsion of the right testis for 2 hours. The animals in groups A and B were treated by gavage with tadalafil at the dose of 0. 5 mg per kg per day, those in group C at 2 mg per kg per day, and those in group D with saline at the same dose. After 3, 7, and 14 days of treatment, the torsioned testes were harvested for evaluation of the superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the testis tissue. The pathological changes in the testis were observed under the light microscope., Results: At 3, 7, and 14 days, the SOD activity was (254.46 +/- 7.43), (278.49 +/- 8.33), and (317.99 +/- 3.31) nU/mg prot in group B, and (277.12 +/- 8.80), (309.40 +/- 2.14), and (320.39 +/- 4.72) nU/mg prot in group C, all obviously higher than in D ([223.21 +/- 4.65], [231.45 +/- 4.16] and [248.28 +/- 5.74] nU/mg prot), while the MDA content was lower in the former two groups than in the latter. At 3 and 7 days, the SOD activity was significantly higher and the MDA level significantly lower in group C than in B (both P < 0.01) , while at 14 days, neither showed any remarkable differences between the two groups (P > 0.05). No obvious histopathological change was observed in the testis tissue of group A. At 3 and 7 days, pathological examination of the testis tissue revealed significant differences in the number of seminiferous epithelial layers, testicular histological score, and seminiferous tubule diameter in group B (P < 0.01), but the three indexes at 14 days in group B and at 7 days in group C exhibited no remarkable differences from those at 14 days in group A., Conclusion: Tadalafil can alleviate testicular ischemia-reperfusion injury following testis torsion/detorsion in a time- and dose-dependent manner.

In order to breed and spread a new cultivar of Curcuma wenyujin, the C. wenyujin germplasm resources were investigated in authentic regions. Better varieties were chosen by comparing the yield, economic characters and quality differences between different cultivars. The results showed that the character of new selected cultivar was stable, the yield of zedoary, turmeric and curcuma was reached 313.7, 177.9, 91.2 kg per 667 m2, respectively, it increased 11.6%, 10.2%, 14.2% comparing with farmer varieties. The volatile oil contents in zedoary and turmeric was 4.0%, 3.0%, respectively. The target ingredients (germacrone) content was stable. It is demonstrated that the new cultivar "Wenyujin No. 1" has value for extension at authentic regions.

Titanium (Ti) based spinal fusion cages are frequently used in the clinics for the treatment of spinal degeneration and related diseases, however, their further clinical application is generally harassed by several drawbacks such as stress shielding, non-biodegradability and additional bone grafting procedure. Our earlier work has demonstrated the efficacy of a biodegradable macro-porous polycaprolactone-tricalcium phosphate (PCL-TCP) composite scaffold in promoting bony tissue ingrowth as well as its ability to sustain mechanical loads upon implantation into an orthotopic defect site. In this study, we investigated the use of PCL-TCP scaffold as an autograft-free spinal fusion cage in a preclinical sheep model over 12 months, and compared the fusion efficacy against Ti cages incorporated with autografts. Results showed that despite PCL-TCP scaffold as an autograft-free cage attaining a slower fusion rate at early stage (6 month), it achieved similar degree of spinal fusion efficacy as Ti cages aided with autograft at 12 month post-operation as evidenced by the radiographic and histological evaluation. PCL-TCP cages alone demonstrated better bone ingrowth with 2.6 fold higher bone/interspace ratio (B/I) and more homogeneous bone tissue distribution compared with that of the Ti cages (88.10  ±  3.63% vs. 33.74  ±  2.78%, p < 0.05) as seen from the histological and micro-CT analysis. Moreover, besides the bone tissue ingrowth, a quantitative approach was illustrated to accurately evaluate the osteointegration of fusion cage with surrounding bone tissue, and showed a 1.36 fold higher degree of osteointegration occurred in PCL-TCP cage group than Ti cage group (CS/PC: 79.31  ±  3.15% vs 58.44  ±  2.43%, p < 0.05). Furthermore, biomechanical analysis showed comparable mechanical strength of fused segments in both groups in terms of the range of motion and stiffness at 12 month (p > 0.05). The degradation profile of the PCL-TCP cages was noted to increase in tandem with new bone ingrowth into the pores, while maintaining good structural integrity necessary for supporting the spinal interbody segments. Therefore, with the better osteointegration, more bone tissue ingrowth as well as its favorable biodegradable and radiolucent properties, PCL-TCP cage has been demonstrated to be a promising candidate as an autograft-free fusion cage for clinical application., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Suspension cultures cell of Sorbus aucuparia (SASC) was used as materials, the changes of physiological and biochemical indexes of SASC after treatment with yeast extract (YE) were detected, and the synthetic mechanism of secondary metabolites in SASC treated with YE was preliminarily explored. The results were as follows: under the assay conditions, SASC was induced to synthesize five biphenyl compounds, and these compounds content changed differently with induction time prolonging; YE treatment inhibited cell growth, the culture medium pH was gradually reduced after treatment; water-soluble protein content showed a trend of slow decline, which was significantly increased in YE treatment group (YE group) compared with the control group (CK group), the maximum relative content was 147.76% in contrast with CK group; both YE group and CK group were extracellular Ca2+ flow influx, but the YE group flow was significantly slow than CK group. The results indicate that YE induced the cells in a stress state, which was not conducive to the growth of cells and forced the cells to synthesize biphenyl compounds against external stress; water-soluble protein may serve as intracellular enzymes involved in the synthesis of compounds regulation; Ca2+ may as signal molecule mediate cell signal transduction respond to YE stress.

Single nucleotide polymorphisms (SNP) is an important molecular marker in traditional Chinese medicine research, and it is widely used in TCM authentication. The present study created a new genotyping method by combining restriction endonuclease digesting with melting curve analysis, which is a stable, rapid and easy doing SNP genotyping method. The new method analyzed SNP genotyping of two chloroplast SNP which was located in or out of the endonuclease recognition site, the results showed that when attaching a 14 bp GC-clamp (cggcgggagggcgg) to 5' end of the primer and selecting suited endonuclease to digest the amplification products, the melting curve of Lonicera japonica and Atractylodes macrocephala were all of double peaks and the adulterants Shan-yin-hua and A. lancea were of single peaks. The results indicated that the method had good stability and reproducibility for identifying authentic medicines from its adulterants. It is a potential SNP genotyping method and named restriction endonuclease digest - melting curve analysis.