A LAMP-Taqman rapid detection system for porcine pseudorabies virus(PRV)was de-veloped based on LAMP and quantitative PCR.LAMP primers were designed for PRV conserved sequences,and the loop primer modified by the fluorescent quenching group was used as the Taq-man probe.The composition optimization,specificity,sensitivity and repeatability of the LAMP-Taqman system were tested using positive samples and recombinant plasmid as templates.Thirty-eight samples of pork swabs were tested with the commercial LAMP detection kit in parallel to verify the actual detection effect of the LAMP-Taqman detection system.The results showed that the optimal final concentration of each component was as follows:PRV-FIP/BIP 0.8 μmol/L,Bst DNA polymerase 0.7 U/μL,Taq DNA polymerase 0.24 U/μL,dNTPs 1.6 mmol/L,MgSO47.2 mmol/L.This system had good specificity and did not cross-react with other virus samples.The linear correlation coefficient of gradient samples was 0.995,the coefficient of variation of repeatable tests was less than 3.000％,and the minimum detection limit could reach 2.81 ×102 copies/μL.The test results of the actual swab samples were consistent with the commercial isothermal fluores-cence detection reagents.In conclusion,the PRV detection system established by LAMP-Taqman method in this study is specific,sensitive,stable and accurate,and is a reliable technical method suitable for the accurate detection of porcine PRV.

This article reviewed the research status, application forms and specific implementation steps of multi-component exercise, and the application of multi-component exercise in elderly patients with sarcopenia. This article analyzed the necessity of multi-component exercise, the insufficiency and prospect of multi-component exercise in elderly patients with sarcopenia, and provides certain reference significance for medical personnel to carry out multi-component exercise intervention in elderly patients with sarcopenia in China.

Objective    To assess the early outcome of transapical transcatheter aortic valve replacement (TAVR) for patients with aortic insufficiency. Methods    The patients with aortic valvular disease who underwent transapical TAVR from October 2020 to October 2022 in the Department of Cardiac and Vascular Surgery, the First Affiliated Hospital of Anhui Medical University were enrolled in the current retrospective study. The patients with aortic stenosis were assembled in a group A, and the patients with aortic insufficiency were assembled in a group B. The improvements of heart function and complications were assessed for the two groups. Results    A total of 56 patients were enrolled, including 32 males and 24 females with an average age of 73.34±5.10 years. There were 31 patients in the group A and 25 patients in the group B. There was no statistical difference between the two groups in the age, gender, height, weight, hypertension, coronary artery disease, peripheral vascular disease, chronic obstructive pulmonary disease, renal disorder or classification of heart function (P>0.05). Also, there was still no statistical difference in the rate of permanent peacemaker implants, emergent open surgery, valve re-implants, or perivalvular leakage (P>0.05). After TAVR, the left ventricular diastolic diameter, left ventricular ejection fraction, complicated moderated mitral and tricuspid regurgitation were significantly improved in both groups compared with preoperative findings (P<0.05); however, there was no statistical difference in these parameters between groups (P>0.05). Conclusion    Interventional valve (J-Valve) in the treatment of patients with aortic insufficiency through transapical TAVR significantly improves cardiac function and reduces functional valve regurgitation.

Metabolomics, which mainly studies the metabolite components of organisms, tissues, cells and their dynamic changes, is an emerging omics technology following genomics and proteomics. Metabolites are the final products of cellular regulation, and the concentration of metabolites is considered to be the ultimate response of a biological system to genetic or environmental changes. Secondary metabolites with chemical diversity are widely present in living organisms, thus accurate quantification of secondary metabolites through appropriate analytical platforms is an important task of metabolomics. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the most commonly used method for the detection of metabolites, providing a basis for the wide application of plant secondary metabolites. This review summarizes the advances of using LC-MS/MS techniques for the detection of phytohormone, folic acid, flavonoids and other secondary metabolites.
Chromatography, Liquid/methods* ; Tandem Mass Spectrometry/methods* ; Metabolomics/methods* ; Plants ; Proteomics

Objective:To examine 2019 novel coronavirus (2019-nCoV) RNA in clinical specimens of COVID-19 patients in Anhui province, and provide evidence for laboratory diagnosis of COVID-19 and risk assessment of clinical specimens.Methods:ORF1ab gene and N gene of 2019-nCoV were detected by real-time fluorescence RT-PCR in 466 clinical specimens of 197 COVID-19 cases. Chi-square test was used to analyze the differences in positive rates of specimens with clinical classification and time of onset.Results:The positive rates of 2019-nCoV in throat swab, sputum, serum, blood sample were 88.83%, 94.67%, 6.78% and 5.08%. The positive rate for 2019-nCoV RNA in throat swabs and sputum differed significantly ( χ2=8.994, P=0.003) in common cases during 7 days after illness onset. Conclusions:The positive rate of RNA in sputum was higher than throat swabs. 2019-nCoV RNA was detected in serum and blood specimens of COVID-19 cases. There was a risk of serum and blood specimens for transmission of COVID-19.

Objective To investigate curative efficacy of transurethral endoscopic surgery for foreign body-induced lower urinary tract injury. Methods A retrospective case series study was conducted to analyze the clinical data of 31 patients with foreign body-induced lower urinary tract injury admitted to Henan Provincial People's Hospital from January 1998 to January 2018. There were 27 males and four females, aged 6-68 years [(27. 1 ± 16. 3)years]. Foreign bodies were located at the bladder in 12 patients, at urethra in 16, and at bladder and urethra in three. The foreign bodies were placed into the body by patients themselves in 23 patients, by other people in one, iatrogenic implantation in six, and by unknown approach in one. All patients underwent urethral surgeries, and individualized treatment plan was formulated according to the specific conditions such as the foreign body type, size, shape, location,activity and combined injury. The success rate, operation time and hospitalization time were recorded. The sexual function of the married males was evaluated by international index of erectile function-5 (IIEF-5) (IIEF-5 score≥22 points as normal) before and after the operation. The urination after the operation was evaluated by the maximum urinary flow rate ( Qmax) ( Qmax<15 ml/s as abnormal) and the complications were recorded. Results All the patients underwent successful surgeries and the foreign bodies were removed, including 26 patients who underwent transurethral endoscopic surgery alone with success rate of 84％ and five patients underwent open surgery due to failed transurethral endoscopic removal. All patients were followed up for 6-36 months [(27. 0 ± 7. 7)months]. The operation lasted for 20-72 minutes [(42. 0 ± 21. 7) minutes]. The hospitalization time was 2-7 days [(4. 0 ± 1. 7) days]. During the follow-up, 11 married males obtained normal sexual function after surgery [ postoperative IIEF-5:(23. 4 ± 1. 1) points vs. preoperative IIEF-5: (23. 8 ± 0. 9) points]. Twenty-nine patients had normal urination [Qmax=(21. 7 ± 5. 9)ml/s]. Two patients had narrow urinary tract (Qmax<15 ml/s), of whom one patient received regular urethral dilatation and another received resection and anastomosis of urethral stricture, both with satisfactory improvement of urination. Three patients had fever caused by urinary tract infection and recovered within 2 weeks after anti-infection treatment. There were no serious complications such as male sexual dysfunction, complex urethral stricture or septic shock. Conclusions Transurethral endoscopic surgery for foreign body-induced lower urinary tract injury has high success rate, minor impact on the sexual function of male patients and low incidence of complications, which is worthy of clinical practice.

Objective To summarize the epidemiological features,diagnosis and treatment of the penile fractures in single clinical center in recent 25 years so as to improve the treatment level.Methods A retrospective case series study was conducted on 23 patients with penile fractures from January 1993 to June 2017.The average age of patients was 37.78 years (range,21-66 years).Nineteen patients were married and four were not.Fourteen patients had penile fractures in summer,five in winter,three in spring and one in autumn.Three patients had Bachelor's degree or above,while 20 had less than university education.There were nine urban and 14 rural residents.Nineteen patients were caused by violent intercourse,two by violent masturbation,one by oral sex and one without willing explanation.All patients were unilateral rupture of albuginea penis,including five with urethral rupture.No misdiagnosis or missed diagnosis occurred.The time from injury to surgery was average 32 h (1993-2002),10.5 h (2003-2012),and 3.8 h (2013-2017),respectively.Emergency operations were performed using the degloved distal foreskin to repair the albuginea penis.The penile erectile function,urination and complications were recorded.Results All patients were followed up for 6-62 months (mean,26 months).In all patients,morning erection was restored at days 1 to 3 after operation,and normal urination was attained after the removal of urethral catheter.Normal sexual life was restored within 3 months after the operation.Penis elephantiasis combined with poor postoperative skin healing was seen in one patient,and mild penile erection pain was reported by one patient.No obvious erectile dysfunction,abnormal urination or other severe complications were reported.Conclusions The epidemiological features of penile fractures include low incidence,relatively concentrated region and age of onset,diversity of causes,and patients with insufficient education.Penile fractures can be diagnosed based on medical history and body examination.Emergency surgery is the first choice for penile fracture,with satisfactory outcomes and few postoperative complications.

Objective: To investigate the mechanisms of nuclear export signal of androgen receptor (NESAR) in the regulation of androgen receptor (AR) protein expression and stability in prostate cancer.Methods: The green fluorescent protein fusion protein expression vectors pEGFP-AR(1-918aa), pEGFP-NESAR (743-817aa), pEGFP-NAR (1-665aa) and pEGFP-NAR-NESAR, and lysine mutants of NESAR pEGFP-NESAR K776R, pEGFP-NESAR K807R and pEGFP-NESAR K776R/K807R, were transiently transfec-ted into prostate cancer cell line PC3.Fluorescence microscopy, Western blot and immunoprecipitation were used to detect NESAR regulation of androgen receptor stability.Results: Under the fluorescence microscope, NESAR-containing fusion proteins were cytoplasmic localization, and their fluorescence intensities were much weaker than those without NESAR.The expression levels of NESAR-containing fusion proteins were significantly lower than those without NESAR.The half-lives of GFP-NESAR and GFP-NAR-NESAR were less than 6 h, while the expression of GFP and GFP-NAR was relatively stable and the half-life was more than 24 h in the presence of cycloheximide.The expression levels of GFP-NESAR were significantly increased by proteasome inhibitor MG132 treatment in a dose-dependent manner;in contrast, MG132 did not show any significant effect on the protein levels of GFP.When new protein synthesis was blocked, MG132 could also prevent the degradation of GFP-NESAR in the transfected cells in the presence of cycloheximide, while it had no significant effect on GFP protein stability in the parallel experiment.GFP immunoprecipitation showed that the ubiquitination level of GFP-NESAR fusion protein was significantly higher than that of the GFP control.The mutations of lysine sites K776 and K807 in NESAR significantly reduced the level of ubiquitination, and showed increased protein stability, indicating that they were the key amino acid residues of NESAR ubiquitination.Conclusion: NESAR was unstable and decreased the stability of its fusion proteins.NESAR was the target of polyubiquitination and mediated the degradation of its fusion proteins through the ubiquitin-proteasome pathway in prostate cancer cells.Our research provides a new way to regulate the level and/or activity of AR proteins, thus helping us understand the molecular mechanisms of AR degradation and strict control of AR in the progression to castration-resistance.

Ginsenosides (GS)as the main ingredient of ginseng,include dozens of monomer,such as Rg3,Rh2,Rd,Rh1 ,Rg2,etc.GS and various monomers have different anti-hepatoma mechanisms.The main mechanism includes the following aspects,inducing the apoptosis of liver cancer cells,inhibiting the prolifera-tion,invasion and metastasis,reversing the multidrug resistance of liver cancer cells,increasing therapeutic effects by combining with other treatments,improving the immune function of organism and inhibiting hepatic glu-cose production.Different GS monomers play different roles of anti-hepatoma through their respective channels.

Objective To investigate the efficacy and safety of transrectal ultrasound guided transurethral balloon dilation of the prostate (TUDP) for the treatment of benign prostatic hyperplasia (BPH).Methods A total of 23 men with BPH who had undergone TUDP were retrospectively analyzed,including 16 men with indwelling urinary catheters before the operation.During the TUDP,the prostatic apex and membranous urethra were dilated by inner balloon,and the prostatic urethra and bladder neck were dilated by outer balloon.The patients were followed up at the 1st,3rd,6th and 12th month after the operation,and the observation parameters included subjective symptoms,such as international prostatic symptom score (IPSS) and quality of life (QOL) score,and objective parameters,such as maximum flow rate (Qmax) and post-void residual (PVR).Results The operation time was 30-165 min,and the intraoperative hemorrhage volume was 5-50 ml.The IPSS scores at 1st,3rd,6th and 12th month after the operation were 10.4± 4.2,8.7±3.2,9.5±4.6 and 8.3±1.5 respectively,which were significantly decreased in comparison to the IPSS score (22.0±7.2) before the operation (P＜0.05).The QOL scores at 1st,3rd,6th and 12th month after the operation were 2.1±1.1,1.6±1.0,1.8±1.1 and 1.6±1.0 respectively,which were significantly improved in comparison to the QOL score (4.9±0.9) before the operation (P＜0.05).The Qmax at 1st,3rd,6th and 12th month after the operation were (10.5±3.4),(12.4±4.2),(10.9±3.9) and (12.7±4.6) ml/s respectively,which were significantly increased in comparison to the Qmax(1.9± 1.9 ml/s) before the operation (P＜0.05).There were 5,4,2 cases of urinary incontinence at the 1st,2nd and 3rd month after the operation,but they recovered at the 4th month follow-up.Besides,there were 3 cases suffered from epididymitis.Conclusion Transrectal ultrasound guided TUDP is a safe,effective and cheap option for the treatment of BPH,especially for the primary hospitals.