Objective To investigate the effect of recombinant plasmid pshRNA-DNMT3b on expression of DNMT3b mRNA and protein and on the proliferation of bladder cancer T24 cells,and research the function of DNMT3b in the process of bladder tumor formation.Methods There were three groups in this study,which are blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),respectively.T24 cells were cultured routinely and transfected by the recombinant plasmids with lipfectamine 2000.The cells were detected by methods of RT-PCR,western blot and MTT.The varying level of DNMT3b mRNA and expression protein,and the conditions of cellular survival rate were observed.Results The recombinant plasmids were successfully transfected into T24 cell lines.The grey valHe of RT-PCR elctrophoretogram was analyzed by the software of Gel-pro analyzer,the rate of blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),was (99.56±1.24)%,(99.12±1.35)%,(75.77±1.42)%,(44.69±1.05)%and(20.52±0.89)%,respectively.The analytical resuit of western blot image was(99.43±1.28)%,(98.90±1.31)%,(67.83±1.02)%,(43.43±1.05)%and(21.92±0.89)%.There was no statistically difference in survival between blank control and HK(P＞0.05).The group of pshRNA-DNMT3b and other two groups had statistical difference only at the 72th hour and the cell inhibitory growth rate only increase 0.45%.Conclusions The recombinant ptasmid pshRNA-DNMT3b can inhibit the expression of mRNA and protein of DNMT3b effectively.However,it has slight function on inhibiting cell proliferation.

Objective To develop an HPLC method for the determination of mycophenolic acid(MPA), mycophenolic acid glucuronide(MPAG) in plasma.Methods The samples were precipitated with zinc sulphate-methanol solution before injection.Carbamazepine was selected as internal standard,ZORBAX XDB C18 (4.6 mm ×250 mm,5 μm) column was used and the flow rate was 1 mL/min.The mobile phase consisted of methanol-acetonitrile-potassium dihydrogen phosphate buffer solution(gradient elution) .The column temperature was 30℃ and the detective wave length was 254 nm.And then the MPA,MPAG concentration of 32 patients in 7-14 days after renal transplantion were determined.Results The assay was linear within 0.2-50μg/mL for MPA, 2.5-500 μg/mL for MPAG(r>0.999).Absolute recovery rates of MPA,MPAG were more than 80%, the recoveries were between 90%-110%.The intra-day and inter-day RSDs were both lower than 10%.Totally 32 cases of renal transplantion patiens were with mycophenolate mofetil at the dose of 1-1.5 g/d,and MPA in plasma was within the range of 0.32-6.19μg/mL,MPAG in plasma was within the range of 9.52-149.25μg/mL.Conclusion The method is accurate, convenient and rapid, which could be used in the quantitative determination of plasma concentration of MPA,MPAG in renal transplantion patients.

Inevitable ischemia-reperfusion injury (IRI) occurred in organ donation after citizen's death often results in delayed graft function (DGF) after renal transplantation. Although the incidence of DGF had reduced to some extent due to continuous hypothermic machine perfusion (HMP) for renal graft using LifePort, DGF of renal graft remains a major clinical problem. In order to further reduce the incidence of DGF after renal transplantation, renal transplant surgeons should fully understand the role of continuous HMP in renal transplantation, continue to conduct more in-depth basic and applied research. This article mainly summarized the effect of continuous HMP on renal transplantation and its research progress.

Objective To evaluate the effect of donor-derived infection (DDI) on clinical prognosis of kidney transplant recipients. Methods Clinical data of 82 donors from donation after citizen's death and 148 kidney transplant recipients were retrospectively analyzed. According to the culture results of the lavage fluid of donor kidney, all recipients were divided into the lavage fluid culture of donor kidney positive group (positive group, n=92) and lavage fluid culture of donor kidney negative group (negative group, n=56). All recipients were assigned into the DDI group (n=19) and non-DDI group (n=129) according to whether they developed DDI or not. The distribution and composition ratio of positive strains in the lavage fluid of donor kidney were analyzed. The incidence of postoperative infection and other complications was assessed in the recipients. Perioperative conditions of the recipients were statistically compared between the DDI and non-DDI groups. The treatment efficacy and clinical prognosis of DDI recipients were evaluated. Results Among 148 recipients, 92 obtained positive culture results in the lavage fluid of donor kidney. A total of 131 pathogenic strains were isolated, including 41.2% (54/131) of Gram-positive cocci, 48.9% (64/131) of Gram-negative bacilli and 9.9%(13/131) of fungi. Among 148 recipients, 52 cases were infected. And 45% (41/92) and 20% (11/56) of the recipients were infected in the positive and negative group, respectively. Statistical significance was noted between two groups (P=0.002). Surgical site was the most common infection site in 52 infected recipients, followed by the urinary system. Nineteen recipients developed DDI with an incidence rate of 12.8% and fatality of 16%. Compared with the non-DDI recipients, DDI recipients had significantly higher graft loss rate and fatality, and longer postoperative hospital stay (all P < 0.05). Eight cases presented with carbapenem-resistant Klebsiella pneumoniae (CRKP) infection, after treatment with tigecycline and/or polymyxin and carbapenems, 3 cases died, and 3 underwent kidney graft resection. In the other 8 recipients with CRKP infection, 2 cases were treated with ceftazidime-avibactam (CAZ-AVI) alone, 3 treated with CAZ-AVI combined with carbapenems, and 3 initially treated with tigecycline combined with carbapenems followed by CAZ-AVI for salvage treatment. After corresponding treatment, the recipients achieved long-term survival. Conclusions DDI may lead to severe complications, while early specific antibacterial treatment plays a positive role.

Objective To evaluate the clinical effects of small-incision orbicularis-levator fixation technique for monocular double-eyelid blepharoplasty.Methods Data were collected in 56 patients (5 males and 51 females) with monocular double eyelid from January 2013 to January 2014.All patients underwent small-incision orbicularis-levator fixation blepharoplasty.To ensure the symmetry between the two eyelids,extensive study and design might be done before the surgery,including the positon of incisions,the amount of tissue removed,the tightness of sutures fixation during surgery,and so on.Results All patients were successfully operated and were performed postoperative follow-up for 19.3±3.4 months.MRI was taken on their upper eyelids preoperatively and 6 months postoperatively.Before the surgery,the MRI orbicularis oculi muscle thickness were 4.08±0.74 U in the left side and 4.81±1.05 U in the right (t=2.66,P＜0.05);the areas of orbit fat were 84.26±9.75 U2 in the left and 93.85±12.15 U2 in the right (t =3.21,P＜0.05).As for 6 months postoperatively,the MRI orbicularis oculi muscle thickness were 4.07±0.69 U in the left side and 4.37±0.99 U in the right (t=1.46,P=0.15);the areas of orbit fat were 82.88±8.81 U2 in the left and 85.30±11.94 U2 in the right (t =1.21,P=0.23).Fifty-four patients were satisfied with surgery,the eyelid's crease turned narrow in 1 case and disappeared in 1 case.Those 2 cases were adjusted by secondary surgery using the same method.Conclusions Small-incision orbicularis-levator fixation blepharoplasty has a high success rate,with acceptable bilateral symmetry and slight scar.

Objective To evaluate the clinical efficacy of en-bloc kidney transplantation from infantile organ donation after citizen's death to adult recipients. Methods Clinical data, surgical approach, use of immunosuppressive agents and follow-up of two adults undergoing kidney transplantation from infantile donor organs were retrospectively analyzed. Relevant literature review was performed. Results One male recipient was diagnosed with primary diseases of chronic renal lesions and renal failure. After kidney transplantation, the recipient obtained favorable recovery of kidney function. The grafted kidney was gradually increased in size. During the final follow-up (10 months after surgery), the serum creatinine level was measured as 84 μmol/L. The other female recipient was diagnosed with renal failure accompanied with uremia. The recipient died from heart failure complicated with severe pulmonary infection at postoperative 23 d. No vascular complications occurred in either recipient. Conclusions Kidney transplantation from infantile donor organs to adult recipients yields favorable clinical efficacy and the grafted kidney is significantly increased in size during the early stage. Precise intraoperative manipulation contributes to preventing the incidence of arterial embolism of the donor kidney and other postoperative complications.

Purpose: The causal relationship between the incidence and prognosis of chronic kidney disease (CKD) and serum testosterone levels in patients is not yet fully understood. This study aims to use the National Health and Nutrition Examination Survey (NHANES), a large-scale nationally representative sample, to investigate the relationship between CKD and testosterone. Materials and Methods: This study included six NHANES cycles for linear regression analysis, verified by multiple imputation methods. Stratified analysis and subgroup analysis were used to demonstrate the stability of CKD’s effect on testosterone. Furthermore, we used Kaplan-Meier plots and log-rank tests to evaluate differences in survival rates between CKD male patients with low and normal levels of testosterone. Results: From a total of 71,163 subjects, the cohort selected 28,663 eligible participants. Results showed that CKD patients had testosterone levels 28.423 ng/mL (24.762, 32.083) lower than non-CKD patients. The results of multiple imputations (β=27.700, 95% confidence interval: 23.427, 31.974) were consistent with those of linear regression analysis, and the numerical match was good. Stratified regression analysis, and subgroup analysis results showed that CKD had a significant impact on testosterone at different dimensions. Kaplan-Meier plots showed significantly reduced survival rates in low testosterone CKD male patients (p<0.0001). Conclusions The results of this big data analysis suggest that there may be a two-way risk between low levels of testosterone and CKD. The testosterone levels of CKD patients were significantly lower than those of the non-CKD population, and CKD patients with low testosterone levels had poorer prognoses. These results suggest that correcting testosterone levels in a timely manner can have preventive and therapeutic effects on the progression of CKD.

Objective To investigate clinical characteristics and risk factors of different stages of urinary tract infection after kidney transplantation.Methods Clinical data of 209 kidney transplant recipients were retrospectively analyzed.According to time points of postoperative follow-up,all recipients were divided into 3 stages:within 1 month post-kidney transplantation,1-6 months post-kidney transplantation,and 7-12 months post-kidney transplantation.The incidence of urinary tract infection,urine culture results of recipients with urinary tract infection and drug resistance characteristics of common pathogens during different stages after kidney transplantation were analyzed.The strains of patients with recurrent urinary tract infection were identified.The risk factors of urinary tract infection and the effect of urinary tract infection on renal allograft function were analyzed.Results The urinary tract infection rate was 90.0％in the first stage,49.3％in the second stage and 22.5％in the third stage.The urinary tract infection rates of male recipients undergoing living-related organ donation in the second and third stages were lower than those of female recipients(both P＜0.05).Urine culture test yielded positive results in 60 cases,and 84 strains of pathogenic bacteria were detected,mainly Gram-negative bacteria,among which Klebsiella pneumoniae accounted for the highest proportion.Sixty-six recipients had recurrent urinary tract infection,and the detected pathogens included Klebsiella pneumoniae,Escherichia coli and Candida glabrata,etc.Univariate analysis showed that postoperative use of antithymocyte globulin was the risk factor for urinary tract infection in the first stage.Preoperative urinary tract infection and donor type were the risk factors for urinary tract infection in the second stage.Gender and age of the recipients were the risk factors for urinary tract infection in the third stage.Multivariate analysis revealed that postoperative use of antithymocyte globulin was the risk factor for urinary tract infection in the first stage.Gender and age of the recipients were the risk factors for urinary tract infection in the third stage(all P＜0.05).In the third stage,65 cases were cured and 38 cases were not cured.In the treated recipients,the serum creatinine level and white blood cell count were decreased after corresponding treatment than those before treatment(both P＜0.05).Conclusions Gram-negative bacteria are the main pathogens of urinary tract infection in kidney transplant recipients,and drug resistance is relatively high.Postoperative use of antithymocyte globulin,female and old age are the risk factors for urinary tract infection in kidney transplant recipients.

Objective:To analyze the clinical characteristics of acute and chronic gastrointestinal bleeding in patients with end-stage renal disease(ESRD)after kidney transplantation,to improve the understanding of the causes,diagnosis,treatment and prevention of this complication,and to improve the management of patients with gastrointestinal bleeding after kidney transplantation.Methods:The clini-cal,imaging and pathological data of patients with gastrointestinal bleeding after kidney transplantation in the Department of Urology of The First Affiliated Hospital of Anhui Medical University from August,2015 to December,2020 were collected.The etiology,early clinical manifestations,abnormal laboratory tests and examinations,treatment procedures,late prevention and treatment measures and outcomes of gastro-intestinal bleeding were retrospectively studied,and the relevant literature was summarized and reviewed.Results:A total of 17 patients were included in this study.Nine patients had chronic small amount of bleeding,hemoglobin gradually decreased,melena and fecal occult blood positive in the early stage,and the general condition was good,vital signs were stable,and were cured by drug treatment.Gastroscopy showed small ulcers with active bleeding foci in 2 cases,and the bleeding was stopped by titanium clips,and the prognosis was good.Gastroscopy showed that the anterior wall longitudinal ulcer at the junction of gastric antrum body was not effective in 1 case,and the small branch of right gastroepithelial artery was embolized,and the patient recovered and discharged after 2 weeks.Gastroscopy showed deep pit ulcer at the lesser curvature of gastric antrum in 1 patient,who underwent distal gastroduodenal artery emboliza-tion and had a good prognosis.Gastroscopy showed huge multiple ulcers in the stomach and duodenal bulb in 2 patients,who underwent subtotal gastrectomy and partial duodenectomy,duodenal stump exclu-sion and remnant gastrojejunostomy.One patient recovered and was discharged,and the other patient died of rebleeding on the 12th day after surgery.Two cases of diverticulum underwent surgical resection of diverticulum,and the prognosis was good.Conclusion:The onset of gastrointestinal hemorrhage in kidney transplant patients is insidious,and the condition is acute or slow,which can cause different de-grees of damage to the patient and the transplanted kidney.Active prevention,early diagnosis,timely drug treatment,if the effect is not good,decisive endoscopic titanium clip hemostasis,transvascular in-terventional embolization,and even surgical treatment can minimize the harm of gastrointestinal bleeding.