The objective of present study was to develop a simple and reliable voiding spot assay (VSA) system to evaluate the lower urinary tract function of mice, and to establish it as a standardized protocol. Ultraviolet (UV) light was used to screen out the filter paper without autofluorescence and with optimal urine diffusion properties. Next, the appropriate wavelength of UV was determined based on the quality of the photographic image of urine spots on the filter paper. To confirm that the urine stain area on the filter paper was correlated with the amount of urine, a volume-area standard curve was constructed. The utility of this VSA system was validated using female wild-type C57BL/6J mice aged 12-13 weeks, and the data generated under identical procedural settings were compared among laboratories. Furthermore, this VSA system was employed to analyze the changes in voiding patterns in mice with urinary tract infections or transportation stress. No. 4 filter paper with a thickness of 0.7 mm was identified as the most suitable material for VSA, exhibiting no autofluorescence and facilitating optimal urine diffusion. The filter paper retained its integrity during the assay, and there was a linear correlation between urine volume and stained area under 365 nm UV light. Utilizing this VSA system, we determined that female wild-type C57BL/6J mice produced approximately 695.8 μL total urine and 5.5 primary voiding spots (PVS) with an average size of 126.4 μL/spot within 4-h period. Over 84% of PVS volumes ranged from 20 to 200 μL. Notably, PVS volumes of mice were similar across different laboratories. Mice with urinary tract infections or transportation stress exhibited significant changes in VSA parameters, including increased voiding frequency, PVS number, and decreased PVS volume. Therefore, this VSA system can be used to evaluate the urinary function of normal mice, as well as those with urinary tract infection or transportation stress.
Mice ; Female ; Animals ; Urodynamics ; Mice, Inbred C57BL ; Urination ; Urinary Bladder ; Urinary Tract Infections

Objective To analyze the risk factors of early acute kidney injury (AKI) after liver transplantation from donation after cardiac death(DCD) donor liver. Methods Clinical data of 184 donors and recipients undergoing liver transplantation from DCD donor liver were retrospectively analyzed. According to the incidence of early AKI, all participants were divided into the AKI and non-AKI groups. The patients in the AKI group were subject to AKI staging. Baseline data, preoperative, intraoperative and postoperative related parameters were statistically compared between two groups. The cumulative survival rate and clinical prognosis of patients in non-AKI group and AKI group with different staging were statistically analyzed by Kaplan-Meier curve analysis. Results Among 184 patients, 68 cases (37.0%) presented with early AKI after liver transplantation including 31 stage 1 AKI, 26 stage 2 AKI and 11 stage 3 AKI, mainly occurring within postoperative 3 d. Univariate analysis revealed that preoperative levels of albumin <35 g/L, preoperative levels of serum sodium ≤137 mmol/L, operation time>7.5 h, intraoperative hemorrhage volume>3 000 mL, intraoperative red cell infusion volume>15 U and intraoperative urine amount ≤100 mL/h were the risk factors of early AKI after liver transplantation (all P<0.05). Multi-variate Logistic regression analysis demonstrated that intraoperative red cell infusion >15 U was an independent risk factor of early AKI after liver transplantation [odds ratio(OR) 1.061, 95% confidence interval(CI)1.008-1.118,P=0.024].Result of Kaplan-Meier survival curve suggested that the cumulative survival rate was gradually reduced along with the aggravation of AKI with statistical significance (all P<0.05). Conclusions The incidence of early AKI following liver transplantation is relatively high. The severity of early AKI is intimately correlated with the short- and long-term prognosis of the recipients. A large quantity of intraoperative red blood cell infusion is an independent risk factor of AKI.

Objective To explore the relationship between WT1 and prognosis of patients with acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL), and to evaluate the possibility of WT1 as a potential marker for monitoring the minimal residual disease (MRD). Methods Bone marrow mononuclear cells from 58 patients with primary AML, 32 patients with primary ALL, 40 patients with AML-complete remission (CR), 28 patients with ALL-CR and 31 patients with trilineage hyperplasia (control group) were collected. Real-time fluorescent quantitative PCR method was used to detect the expression of WT1 in all patients. The expression threshold of WT1 in each group was established. WT1 copy number/ABL copy number ratio×100%denotes the relative expression level of WT1 gene. Results Median relative expression level of WT1 in the control patients was much lower than that in primary AML patients [0.026%(0-0.240%) vs. 20.880 % (3.550 %-48.500 %), Z=-7.74, P<0.000 1]. Relative expression level of WT1 between AML-CR patients [0.102%(0-5.380%)] and primary AML patients had significant difference (Z=-8.34, P<0.000 1). Moreover, the relative expression level rate of the first course in AML patients with higher WT1 expression level (>20.880 %) was 60.7 % (17/28), while the CR rate was 76.7 % (23/30) in those with lower WT1 expression. WT1 expression was increased dramatically in recurrent AML patients. Relative expression level of WT1 was significantly higher in primary ALL patients [0.350 % (0.021 %-10.780 %)] compared with that in the control group Z=-2.58, P<0.05. There was no significant difference in relative expression level of WT1 between ALL and ALL-CR patients [0.038 % (0-2.800 %), P=0.065]. Conclusion WT1 expression level in AML patients is relatively high, which could be used as an effective index of prognosis evaluation and MRD monitoring for AML patients, but not for ALL patients.

Objective To investigate the relationship of central venous pressure (CVP) and organ function in early period after orthotopic liver transplantation (OLT).Methods A retrospective study was conducted on 111 patients who underwent OLT.According to the value of mean CVP after OLT,all patients were divided into three groups:low CVP group (CVP＜8 rnmHg,1 rnmHg =0.133 kPa),normal CVP group (CVP 8-12 mmHg) and high CVP group (CVP ＞12 mmHg).Meanwhile,According to whether the CVP dropped below 8 mmHg or not in the past 48h after surgery,all patients were divided into two groups.Results There were significant differences in serum total bilirubin,serum creatinine and serum lactate among low,normal,and high CVP groups (P＜0.05).The time of vasoactive agent,fluid balance,time of mechanical ventilation and incidence of acute kidney injury in groups with CVP not dropped below 8 rnmHg were higher than those in groups with CVP dropped below 8 mmHg (P＜0.05).Conclusion CVP was associated with liver,kidney,lung function and lactate.Controlling a lower CVP can significantly shorten the time of mechanical ventilation and reduce the incidence of acute kidney injury after OLT.

Objective To summarize the clinical efficacy of liver transplantation from donation after cardiac death (DCD). Methods Clinical data of both the donors and recipients (n=182) undergoing liver transplantation from DCD were retrospectively analyzed. According to the type of primary diseases, 182 recipients were divided into the benign group (n=135) and hepatocellular carcinoma (liver cancer) group (n=47). Perioperative conditions, 1- and 3-year survival rate of the recipients were statistically compared between two groups. Clinical prognosis and the incidence of postoperative complications of the recipients were summarized. Postoperative complications mainly included early allograft dysfunction (EAD), vascular complications, acute kidney injury (AKI), pulmonary infection, acute rejection, cytomegalovirus (CMV) infection and billiary tract complication. Results No statistical significance was identified in the anhepatic phase, operation time and length of intensive care unit (ICU) stay between two groups (all P>0.05). The 1-year survival rates of the 182 recipients and grafts were 93.1%, and 84.9% for the 3-year survival rates. In the benign group, the 1- and 3-year survival rates of the recipients were 92.5% and 88.1%. In the liver cancer group, the 1-year survival rate of the recipients was 95%, 91% for the disease-free survival rate, and 78% for the 3-year survival rate, respectively. No statistical significance was noted in the overall survival rate of the recipients between two groups (P=0.879). In terms of postoperative complications, billiary tract complications occurred in 26 patients, vascular complications in 14, AKI in 34, pulmonary infection in 22, acute rejection in 11, EAD in 11 and CMV infection in 10. The incidence of postoperative billiary tract complications in patients with T-tube insertion was significantly lower than that in their counterparts without T-tube insertion (8% vs. 19%, P<0.05). Conclusions Liver transplantation from DCD is an efficacious treatment for end-stage liver diseases and liver cancer, which yields relatively high short-term clinical efficacy.

Background:Acute myeloid leukemia (AML) with t(8;21) is a heterogeneous disease. Identifying AML patients with t(8;21) who have a poor prognosis despite achieving remission is important for determining the best subsequent therapy. This study aimed to evaluate the impact of Wilm tumor gene?1 (WT1) transcript levels and cellular homolog of the viral oncogenev?KIT receptor tyrosine kinase (C?KIT) mutations at diagnosis, andRUNX1?RUNX1T1 transcript levels after the second consolidation chemotherapy cycle on outcomes.
Methods:Eighty?eight AML patients with t(8;21) who received chemotherapy only or allogeneic hematopoietic stem cell transplantation (allo?HSCT) were included. Patients who achieved remission, received two or more cycles of consolidation chemotherapy, and had a positive measureable residual disease (MRD) test result (deifned as<3?log reduction inRUNX1?RUNX1T1 transcript levels compared to baseline) after 2–8 cycles of consolidation chemotherapy were recommended to receive allo?HSCT. Patients who had a negative MRD test result were recommended to receive further chemotherapy up to only 8 cycles.WT1 transcript levels andC?KIT mutations at diagnosis, andRUNX1?RUNX1T1 transcript levels after the second consolidation chemotherapy cycle were tested.
Results:Patients who had aC?KIT mutation had signiifcantly lowerWT1 transcript levels than patients who did not have aC?KIT mutation (6.7%±10.6% vs. 19.5%±19.9%,P<0.001). LowWT1 transcript levels (≤5.0%) but notC?KIT mutation at diagnosis, a positive MRD test result after the second cycle of consolidation chemotherapy, and receiv?ing only chemotherapy were independently associated with high cumulative incidence of relapse in all patients (hazard ratio [HR]=3.53, 2.30, and 11.49; 95% conifdence interval [CI] 1.64–7.62, 1.82–7.56, and 4.43–29.82;P=0.002, 0.034, and<0.001, respectively); these conditions were also independently associated with low leukemia?free survival (HR=3.71, 2.33, and 5.85; 95% CI 1.82–7.56, 1.17–4.64, and 2.75–12.44;P<0.001, 0.016, and<0.001, respectively) and overall survival (HR=3.50, 2.32, and 4.34; 95% CI 1.56–7.82, 1.09–4.97, and 1.98–9.53;P=0.002, 0.030, and<0.001, respectively) in all patients.
Conclusions: Testing forWT1 transcript levels at diagnosis in patients with AML and t(8;21) may predict outcomes in those who achieve remission. A randomized study is warranted to determine whether allo?HSCT can improve prog?nosis in these patients.

Objective To investigate the effect of fatty acid synthase (FASN)on apoptosis in pancreatic cancer cell PANC-1 and the possible molecular mechanism.Methods Annexin V/FITC and flow cytometry were performed to detect the expression of FASN in pancreatic cancer PANC-1 after C75 treatment and the change of apoptosis in pancreatic cancer cell PANC-1 treated with C75.Quantity reverse transcriptase polymerase chain reaction (RT-PCR)and Western blot were used to measure the protein and RNA expressions of Caspase-3,bcl-2 and FASN.Results Inhibited by C75,the activity of FASN in pancreatic cancer cell PANC-1 was significantly decreased.Meanwhile,PANC-1 showed an increased apoptosis level in a dose-dependent manner (P < 0.05 ). Furthermore,after C75 inhibited FASN in pancreatic cancer cells,the protein and RNA expressions of Caspase-3 significantly increased (P <0.05)whereas the level of Bcl-2 reduced (P <0.05).Conclusion FASN is involved in the process of apoptosis in PANC-1 via Bcl-2 and Caspase-3.Therefore,FASN will provide a new target for the treatment of pancreatic cancer and generate better treatment efficacy.

Objective The aim of this study is to demonstrate the diagnostic effect of brainstem evoked potential for pancreatic encephalopathy in rats with severe acute pancreatitis. Methods Sixty male Sprague-Dawley (SD) rats were randomly divided into two equal groups: a sham-operated (SO) group and a severe acute pancreatitis (SAP) group. Each group was evaluated at 3, 6, and 12 h during the experiment. To detect the brain stem evoked potential change at different time points. The ultrastructure of brain tissue was observed by transmission electron microscope (TEM). The expressions of the apoptosis-related proteins Bcl-2, Bax and caspase-3 were observed using immunohistochemical and Western Blot technique. Results In SAP group, congestion, edema, inflammatory cell infiltration, mitochondrial swelling and cell apoptosis were apparent. Compared with SO group, the brain stem evoked potential in severe acute pancreatitis group was obviously reduced in SAP group. Compared with SAP group, the expressions of Bcl-2 have increased, whereas the expressions of Bax and caspase-3 have decreased in SO group significantly ( <0.05) . Conclusion Brain stem evoked potential is a sensitive method in detection of rat brain damage. The results showed that the consistency and the damage degree of rats may be important clinical diagnostic index of pancreatic encephalopathy.

Objective Estahlished the method to detect different transcripts of EVI1 gene expression with quantitative PCR and study the expression patterns of EVI1 gene in different leukemia groups to investigate the association between EVI1 gene expression and the incidence and prognosis of leukemia.Methods 60 cases acute myeloid leukemia (AML) and 9 cases normal control were detected in the study,37 cases were male and 32 cases were female,age 10-70 years,median age 42 years,M3 36 cases,M2 16 cases and M4 8 cases according to FAB classification criteria,control samples of nine cases were normal healthy people.Using the quantitative PCR (Taq Man probe) to detect the expression of different transcripts of EVI1 gene.The t test was used to detect the expression difference among different leukemia groups.Results ABL gene was used as internal reference,relative changes of EVI1 gene expression level were detected by EVI1/ABL.In all the control patients,EVI1 gene of different transcription of this expression were detected,expression level of EVI1 gene different transcription was significant with the difference (P ＜ 0.05),transcription 2 and 5 (the same primers) were the lowest,followed for transcription 1 and 6,expression of transcription 3 was the highest.The expression levels of transcripts 2 and 5,1,6,3 were nagative,0.005,0.050 and 0.512 respectively in healthy control samples.In addition,the EVI1 gene expression was negatively correlated with expression of the fusion gene AML-ETO and CBFB-MYH11 in AML.Conclusion The study established a stable,fast and accurate method to detect the expression of EVI1 gene.

Objective To delineate the potency of YC-1 on the proliferation,apoptosis,cell cycle and the protein expression of Caspase-3,-8,-9 in U937 and THP-1 leukemia cell lines.Methods MTT assay was performed to detect proliferation.Flow cytometry was used to measure the apoptosis and cell cycle.The expression of Caspase-3,-8 and-9 were detected by Western blot.Results The MTT assay showed that cell proliferation was inhibited in a concentration-dependent manner in 1.0,3.0,10.0 μmol/L YC-l-treated U937 and THP-1 cells.The survival rates for YC-1 after 24 h in U937 cells were (76.5±4.4) ％,(68.7±6.8) ％,(60.9±13.2) ％ respectively and (94.1±1.4) ％,(81.4±2.0) ％,(72.7±3.0) ％ respectively in THP-1 cell,compared with the control group (100 ％),there were significant differences (F =15.870,126.629,P ＜ 0.01).The apoptosis rates for 1.0,3.0,10.0 μmol/L YC-1 after 24 h were (40.7±1.0) ％,(55.6±2.3) ％,(71.8±1.5) ％respectively in U937 cells and (34.6±2.0) ％,(50.3±3.5) ％,(59.6±4.6) ％ respectively in THP-1 cells.With the control group (4.7±1.4) ％,(1.8±1.0) ％,there were significant difference (F =937.229,200.447,P ＜ 0.01).However,there was no significant difference for cell cycle.In addition,Cleaved Caspase-8 and Cleaved Caspase-3 expression after 1.0,3.0,10.0 μmol/L YC-1 treated for 24 h were significantly higher than control,but the expression of Caspase-9 did not appear significant change in U937 cells.As the same concentration and time point,Cleaved Caspase-3 expression increased with no change of Caspase-9 or Caspase-8 in THP-1 cells.Conclusion YC-1 effectively suppress the proliferation with little effect on cell cycle,but induce the apoptosis,have no effect on cell cycle,and the mechanism of apoptosis may be related to the Caspase activation in U937 and THP-1 cell lines.