Objective To investigate the effect of CaMK Ⅱ expression on apoptosis of rat hepatocytes BRL-3A.Methods Rat BRL-3A cells were stable passage were cultured.The CaMK Ⅱ γ protein (LV-CaMK Ⅱ γ group) and CaMK Ⅱ γshRNA (shRNA group) lentiviral expression systems were constructed.The corresponding blank vectors (LV-NC group and shRNA-NC group) and normal saline (CON group) were perfused into the control groups.The expression levels of CaMK Ⅱ,Cyt C and MF proteins were detected by Western blotting,and the apoptosis rate of BRL-3A cells was measured by Tunel method.Results The protein expression of CaMK Ⅱ,Cyt C and AIF in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).The protein expression of CaMK Ⅱ,Cyt C and AIF in shRNA group was significantly lower than that in CON group (P＜ 0.05).There was no significant difference among CON group,LV-NC group and shRNA-NC group (P＞0.05).At the same time point,the apoptosis rate of hepatocytes in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).At the same time point,the apoptosis rate of hepatocytes in shRNA group was significantly higher than in CON group (P＜0.05).There was no significant difference in the apoptosis of hepatocytes among CON group,LV-NC group and shRNA-NC group (P＞0.05).Conclusion The specific CaMK Ⅱ signaling pathway can inhibit the apoptosis of BRL-3A cells,while the enhanced CaMK Ⅱ signaling pathway promotes the apoptosis of BRL-3A cells.

Objective To analyze the follow-up results and clinical characteristics of one case of highly sensitized recipient after combined kidney transplantation and splenic fossa auxiliary heterotopic liver transplantation.Methods This patient was diagnosed as having chronic renal insufficiency in the uremia period 10 years ago,subjected to kidney transplantation 9 years ago,and got renal allograft loss 8 years ago.The recipient was positive for PRA (for class Ⅰ,31％,and for class Ⅱ,63％).Under the general anesthesia,the patient was given combined kidney transplantation and splenic fossa auxiliary heterotopic liver transplantation.The ATG was used for immune induction.Rituximab and plasma exchange were applied to prevent acute rejection.Regular follow-up was done after discharge.Results On the postoperative day (POD) one,ALT was 256 IU/L,AST was 342 IU/L and serum creatinine was 502 μmol/L.On the POD 6,ALT and AST levels were normal and serum creatinine was 141 μmol/L.Serum creatinine increased to 202 μmol/L and the volume of urine reduced on the POD 7.The ultrasound displayed graft size increased slightly,substantial echogenicity enhanced,artery blood flow RI increased to 0.8,suggesting the occurrence of acute rejection.A single dose of Rituximab,intravenous IG,and plasma exchange were given.On the POD 60,serum creatinine was reduced to 131 μmol/L.During a follow-up period of 28 months,imrnunosuppresants were given:Tac + MMF + Pred.FK506 valley concentration was maintained at 6-8 μg/L.The function of the transplanted kidney and liver was normal,and the general conditions were good.Conclusion Combined kidney transplantation and splenic fossa auxiliary heterotopic liver transplantation is safe.Individualized medication and regular follow-up are the important factors for long-term survival of recipients.

Objective To evaluate the safety and efficacy of ex vivo ureteroscopy (ExURS) as means of rendering a donated kidney stone-free in a living related renal transplantation.Methods Clinical data were analysed of ExURS as means of rendering a donated kidney stone-free in a living related renal transplantation and relative literature was reviewed.The ECT results showed that GFR of left and right kidney was 38.7 and 42.3 ml/min respectively.The donor underwent a left laparoscopic donor nephrectomy.Immediately after cold perfusion,ExURS was performed with 4 ℃ ice-cold saline irrigation.Basket extraction and holmium laser lithotripsy was performed.Calculi were fragmented with pneumatic intracorporeal lithotripsy and fragments were removed with forceps.F6 indwelling ureteral stents were kept during transplantation.Urine flowed out immediately after reperfusion of the allograft and the distal ureter appeared edema 2 min later.Routine ureter-bladder wall anti-reflux replantation was done after the resection of the edema part.Results Pyeloscopy was successfully performed.A total of 2 calculi,diameter 8,12 mm,were visualized in donor kidney.The ex vivo treatment time was 30 nin.The warm and cold ischenia time was 60s and 50 min,respectively.There were no intraoperative complications.At a follow-up at 8 months,there was no recurrent calculi formation in the recipient and donor.Conclusion ExURS is technically feasible to render a stone-bearing kidney stone free without compromising ureteral integrity or renal allograft function.

Objective To evaluate the clinical efficacy of adult kidney transplantation from unilateral pediatric donor kidney. Methods Clinical data of pediatric donors (n=10) and adult recipients (n=19) undergoing kidney transplantation were retrospectively analyzed. The changes of renal function, liver function and the maximal diameters of the kidney allografts were compared at 1, 7, 14, 28, 60 d after operation. The short-term survival and incidence of postoperative complications of the recipients were analyzed. Results Ten donors included 6 males and 4 females, aged (7±3) years old, with a body mass index (BMI) of (16.3±3.8) kg/m². All donors were donation after brain death followed by cardiac death. Among 19 recipients, 12 were males and 7 were females, aged (34±12) years old, with a BMI of (20.3±1.3) kg/m².An oblique incision was created in the lower right abdomen of the recipients. The arteries and veins of donor kidney were anastomosed with the external iliac arteries and veins of the recipients. The ureter of donor kidney was anastomosed with the bladder of the recipients. After anastomosis, the kidney was placed and fixed in the right iliac fossa. The serum creatinine and blood urea nitrogen levels of the recipients were decreased at 1 week after kidney transplantation, and restored to normal range at postoperative 2 weeks. All parameters related to liver function were normal after operation. At postoperative 1 month, the maximal diameters of the kidney allografts were (9.5±0.3) cm on average, which basically reached those of normal adults. The 1-year survival rate of 19 recipients was 95%. One recipient died from pulmonary infection after ineffective treatment. Two recipients developed rejection, and 1 recipient experienced urinary system infection, who were healed after corresponding treatment. Conclusions Adult kidney transplantation from unilateral pediatric donor kidney is safe, feasible and effective, which can be utilized to enlarge the source of donor kidneys.

Objective Toinvestigatetherelationshipbetweentheexpressionofstromalcellderived factor (SDF)-1,intercellular adhesion molecule (ICAM)-1 and renal tubular necrosis score in rats with renalischemicreperfusioninjury(IRI).Methods SixtySprague-Dawley(SD)ratswererandomlydivided into operation group and sham operation group with 30 rats in each group. Then each group was divided into 6 subgroups (1 h,6 h,12 h,24 h,48 h or 72 h )according to different time of measurement after operation with 5 rats in each subgroup. Renal IRI mode1 s were built in rats of operation group. The bilateral renal arteries were dissected in rats of sham operation group and then the incision was sutured. The renal function, renal tubular necrosis score and the variation of SDF-1 ,ICAM-1 expression in renal tissues at different time points were measured. The relationship between the expression of SDF-1 ,ICAM-1 in renal tissues and renal tubularnecrosisscoreinratsofoperationgroupwasanalyzedbyPearsoncorrelationanalysis.Results The levels of urea nitrogen (BUN)and serum creatinine (Scr)after operation in each subgroup of operation group were significantly higher than those before operation and those in the corresponding subgroups of sham operation group (all in P<0.05). They increased significantly 12 h after operation and reached the peak at 48 h after operation. The renal tubular necrosis score in operation group increased gradually over time (all in P<0.05 ).The highest renal tubular necrosis score was in the 48 h operation subgroup (P<0.05 ). Compared with those in 1 h operation subgroup,the expression of SDF-1 ,ICAM-1 in rats' renal tissues of 6 h operation subgroup began to increase significantly,and they reached the peak at 48 h after operation and began to drop down at 72 h after operation. The expression of SDF-1 ,ICAM-1 in rats renal tissues in operation group were positively correlated with the levels of BUN,Scr and renal tubular necrosis score at different time points after operation (r=0.614,0.662,0.751;0.640,0.703,0.785;P<0.05).Conclusions Whenrat'srenaltissuedevelops IRI,the expression of SDF-1 ,ICAM-1 ,BUN,Scr and renal tubular necrosis score increased.The expression of SDF-1 ,ICAM-1 are positively correlated with BUN,Scr and renal tubular necrosis score. The increased expression of SDF-1 ,ICAM-1 can serve as an indicator of the severity of renal IRI.

Objective To investigate the diagnosis and treatment regimen for patients with chronic diarrhea after renal transplantation.Methods The clinical data of 353 patients with chronic diarrhea who underwent renal allograft transplantation at Department of Urology of Xijing Hospital from January 2007 to June 2014 with regular follow-up were analyzed retrospectively.The occurrence of chronic diarrhea after renal transplantation was observed,including incidence,time of occurrence,course of disease and complications. The changes in general conditions and auxiliary examination indexes (body mass index,anemia and other auxiliary examination indexes),treatment and prognosis of the patients with chronic diarrhea were recorded. Results Fifteen cases (4.2%) of 353 renal transplant recipients had chronic diarrhea. The time of symptomatic and etiological treatment was (15 ±7 ) d.Two patients died during diarrhea (died from gastrointestinal hemorrhage and sudden death caused by severe hypokalemia respectively)and other patients were recovered.Among the 13 patients,5 cases had good prognosis,2 cases died (both died from pulmonary infection),5 cases suffered from renal allograft dysfunction and 1 case suffered from renal allograft insufficiency during the follow-up.Conclusions The etiology of chronic diarrhea after renal transplantation is complex and the patients should receive symptomatic and etiological treatment.The patients with chronic diarrhea after renal transplantation combined with severe complications have poor prognosis.

Objective To discuss the safety of Rh-positive patients receiving kidney grafts from Rh-negative cadaver donor. Methods On November 29 th 2013, two Rh-positive patients received renal transplantation with kidney grafts from Rh-negative cadaver donor at the Department of Urinary Surgery of Xijing Hospital,the Fourth Military Medical University. The clinical data of the two patients were analyzed retrospectively and the relevant literatures were reviewed. Results The two patients underwent renal transplantation successfully and no hyperacute rejection or acute rejection occured after the surgery.The two patients were followed up for 12 months.The kidney grafts survived well and the patients had no obvious discomfort.Conclusions Through enhanced immunosuppression before operation,sufficient perfusion and close postoperative monitoring,it is safe for Rh-positive patients receiving kidney grafts from Rh-negative cadaver donor.

Objective To evaluate the safety and efficacy of robot﹣assisted laparoscopic living donor nephrectomy. Methods Clinical data of 31 donors and recipients undergoing robot﹣assisted laparoscopic living donor nephrectomy in Xijing Hospital of the Fourth Military Medical University from November 2013 to August 2015 were retrospectively analyzed. Results Donor nephrectomy was successfully performed in 31 cases.The operation time ranged from 110 to 190 min.Intraoperative hemorrhage volume was measured as 20﹣100 ml.The warm ischemia time of the donor kidney was 100 to 160 s.The retained length of renal vein was between 1.8 and 3.0 cm and the length of renal artery was 1.4 to 2.3 cm.In 2 cases,spleen injury occurred during the kidney extraction and was treated with splenorrhaphy.One donor had postoperative hemorrhage,which was treated with hemostasis and anemia correction.Thirty one donors received postoperative follow﹣up for over 6 months.No long﹣term complications were observed.Among 31 recipients,one patient had delayed recovery of renal graft function and the serum creatinine level returned to normal range after treatment at postoperative 1 month.The survival rate of kidney grafts was up to 100%. Conclusions Robot﹣assisted laparoscopic living donor nephrectomy is a safe and efficacious surgical procedure for kidney resection,which possesses the advantages of small trauma,rapid recovery and no influence upon renal function.

Objective To evaluate the effect of hypothermia status in the donors upon the renal graft function after renal transplantation from donation after citizen's death. Methods Thirty-six eligible donors were randomly divided into the normal temperature (body temperature 36.5-37.5 ℃ , n=19) and hypothermia groups (body temperature 34.0-35.0 ℃ , n=17). The matched recipients undergoing renal transplantation were also assigned into the normal temperature (n=38) and hypothermia groups (n=34). Perioperative conditions of the donors and recipients were compared between two groups. And postoperative renal graft function of the recipients were statistically compared between two groups, including the incidence of delayed graft function (DGF) and primary nonfunction (PNF). Results No statistical significance was identified in the perioperative amount of urine volume, serum creatinine (Scr), systolic blood pressure, saturation oxygen, warm ischemia time and cold ischemia time of the donors between two groups (all P>0.05). No statistical significance was noted in terms of the operation time, intraoperative mean blood glucose and intraoperative mean arterial pressure of the recipients between two groups (all P>0.05). Postoperative incidence of DGF of the recipients in the hypothermia group was 6%, significantly lower than that in the normal temperature group (24%) (χ2=4.393, P=0.036). Postoperative incidence of PNF of the recipients was 3% in both the hypothermia and normal temperature groups with no statistical significance (χ2=0.000, P=1). Conclusions The hypothermia status of the donors can significantly reduce the incidence of DGF, whereas exerts no evident effect upon the incidence of PNF in the recipients.

Objective To investigate the clinical characteristics, prevention and treatment of multi-drug resistant organisms (MDROs) infection early after renal transplantation from donation after citizen's death. Methods Clinical data of 166 patients undergoing allogeneic renal transplantation and regular follow-up in Xijing Hospital from November 2011 to September 2016 were retrospectively analyzed. General conditions were statistically compared between the recipients undergoing renal transplantation from donation after cardiac death (DCD) and their counterparts receiving living related donor renal transplantation. The incidence of MDROs infection, onset time, course of diseases, complications, infection site and etiological type were observed. The therapeutic methods and clinical prognosis were summarized. Results The incidence of MDROs infection early after renal transplantation in the recipients undergoing DCD renal transplantation was 14%, significantly higher than 2% in those receiving living related donor renal transplantation, and 13% and 2% for the incidence of delayed graft function with statistical significance (both P<0.05). The incidence of renal graft loss was 8%and 2%, and 5% and 1% for the mortality rate without statistical significance between two groups (both P>0.05). MDROs infection occurred in 11 patients after DCD renal transplantation. The most common infection site was urinary system(n=6) and the most prevalent pathogenic bacterium was Escherichia coli (n=4). All patients infected with MDROs were treated with a sufficient dosage of effective antibiotics according to the outcomes of bacterial culture and drug sensitivity test. Eight patients obtained favorable clinical prognosis, one underwent nephrectomy and two died. Conclusions The incidence of MDROs infection early after DCD renal transplantation is higher than that after living related-donor renal transplantation. Strict donor screening, early detection, intimate monitoring and timely treatment can effectively reduce the risk of MDROs and enhance clinical prognosis.