Objective To construct the lamino acid transporter 1 eukaryotic expression vector of C 57 mouse and to express the gene inNeuro-2atumor cells,and explore the effect of LAT1on proliferation and apoptosis of Neuro-2a cell. Methods The full-length LAT1 cDNA was synthesized by RT-PCR and cloned into pcDNA3.1vector to construct recombinant plasmid.The constructed pcDNA3.1-LAT1vector was verified by Enzyme digestion and sequencing and then transfected intomurine Neuro-2acellsby liposome.The transfected cells were selected with G418 and stably expressed strain was constructed .The expression of LAT1 was detected by RT-PCR and western blot .Proliferation was analyzed by MTT , cell cycle and apoptosis were detected by flow cytometric analysis .Results The full-length LAT1 cDNA was amplified successfully and pcDNA3.1-LAT1eukaryoticvector was constructed successfully .Enzyme digestion and sequencing confirmed the sequence was correct .Neuro-2acells were transfected and Stably expressed strain was constructed successfully.MTT showed that the group of transfected restructuring plasmid could significantly affect Neuro -2a cell proliferation more than the control groups ( P <0.05 ) .From the flowcytometric analysis , LAT1 could promote cell proliferation and inhibit Neuro-2a cell apoptosis.Conclusion LAT1 can express successfully inNeuro-2acells which were transfected with recombinantpcDNA3.1-LAT1plasmid.LAT1 in Neuro-2a cells can promote cell proliferation and inhibit the cell apoptosis which provides a basis for the study of LAT 1.That lays the foundation for studying biological effects of LAT 1.

Cholestasis ; etiology ; Gestational Age ; Humans ; Hydrops Fetalis ; physiopathology ; Infant, Newborn ; Liver Function Tests ; Male ; Tachycardia, Paroxysmal ; complications ; embryology ; physiopathology ; Tachycardia, Supraventricular ; complications ; embryology ; physiopathology

Objective To investigate the expression changes of macrophage migration inhibitory factor( MIF) and C-Jun N-terminal kinase ( JNK) in the liver of rat with abnormal glucose metabolism and explore the pathological mecha-nism of abnormal glucose metabolism with non-alcoholic fatty liver disease ( NAFLD) .Methods Sixty SD rats were ran-domly divided into impaired glucose tolerance (IGT) model group (n=20), type 2 diabetes mellitus (T2DM) model group (n=20), IGT control group (n=10) and T2DM control group (n=10).IGT models were produced by feeding high-fat diet, T2DM models were produced by feeding high-fat diet for 4 weeks and intraperitoneally injected streptozotocin. Liver cell apoptosis was detected by TUNEL staining.The expression of MIF mRNA in liver tissue was determined by real-time PCR.The expressions of MIF, caspase-3, JNK proteins and phosphorylated JNK(p-JNK)in the liver tissue were de-termined by Western blotting.Results Apoptotic cells were obviously increased in the IGT and T2DM groups.Expression of MIF mRNA in the IGT and T2DM groups was markedly higher than that in the control group, respectively (P<0.01). The expressions of MIF, caspase-3, JNK proteins and phosphorylated JNK were markedly higher than that of the control group, respectively (P<0.05 or P<0.01).The expressions of MIF, caspase-3, JNK proteins in the T2DM group were significantly decreased compared with those of the IGT group, while the expression of phosphorylated JNK was significantly higher ( P<0.01) .Conclusions Abnormal glucose metabolism accompanied with NAFLD is probably related to the in-crease of MIF, Caspase-3, JNK and phosphorylated JNK expression.

[ ABSTRACT] AIM:To observe the changes of perilipin and adipose differentiation-related protein ( ADRP) du-ring the development of diabetes mellitus and to explore the effect of perilipin and ADRP on abnormal glucose metabolism with non-alcoholic fatty liver disease ( NAFLD) .METHODS:The rat model of impaired glucose tolerance ( IGT) was in-duced by feeding high-fat diet, and the type 2 diabetes mellitus (T2DM) model was induced by feeding high-fat diet for 4 weeks and intraperitoneally injecting streptozotocin.The morphological change of the liver tissue was observed under optical microscope.The serum contents of perilipin and ADRP were measured by ELISA.The mRNA expression of perilipin and ADRP in the liver tissues was detected by real-time PCR.The protein expression of ADRP in the liver tissues was deter-mined by Western blotting.RESULTS:HE staining showed steatosis in the liver of the rats in IGT group was more serious than that in T2DM group.The biochemical and the pathological processes of rat models were consistent with the clinical feature of related diseases.The serum content of perilipin had no difference among various groups.The mRNA expression of perilipin in IGT group and T2DM group was significantly higher than that in control group.Compared with IGT group, the mRNA expression of perilipin in T2DM group was significantly increased.The serum content of ADRP in T2DM group was significantly lower than that in control group.The mRNA and protein expression of ADRP in model groups was signifi-cantly lower than that in control group.Compared with IGT group, the mRNA expression of ADRP in T2DM group was sig-nificantly reduced.CONCLUSION: The serum content of ADRP plays a role in the development and progression of T2DM.It is negatively correlated with HOMA-IR.NAFLD occurs during progression of abnormal glucose metabolism in-duced by feeding high-fat diet.The development of abnormal glucose metabolism with NAFLD is probably related to the in-creased expression of perilipin and the reduced expression of ADRP.

Objective To evaluate the predictive factors and renal outcomes of idiopathic membranous nephropathy (IMN) in patients with type 2 diabetes (T2DM).Methods In this retrospective study,clinical data of 101 IMN patients with T2DM and 96 patients with diabetic nephropathy (DN) were consecutively collected.Logistic regression was used to assess potential clinical factors indicating IMN and COX regression was employed to analyze risks of IMN in developing to endstage renal disease (ESRD),as compared with that of DN,in patients with T2DM.Results In a multivariate model,age ≥55 years old,presence of nephrotic syndrome,estimated glomerular filtration rate (eGFR) ＞ 60 ml · min-1 · (1.73 m2)-1,duration of diabetes≤5 years and absence of diabetic retinopathy,were associated with IMN,as compared with DN,in patients with T2DM.In T2DM patients presented with nephrotic syndrome,age≥55 years old,eGFR ＞ 60 ml· min1· (1.73 m2)-1,duration of diabetes≤5 years and absence of diabetic retinopathy,were also associated with IMN,as compared with DN.Receiver operating characteristic curve (ROC) showed eGFR 65.5 ml · min-1 · (1.73 m2) 1 was an optimal cutoff in differentiating DN and IMN.DN was associated with 16.8 times as high risk of incident ESRD as compared with IMN in T2DM patients.Conclusions In patients with T2DM,age≥55 years,presence of nephrotic syndrome,early stage of CKD,duration of diabetes≤5 years and absence of retinopathy,may indicate IMN rather than DN.T2DM patients with IMN have much better renal prognosis as compared with DN.

Objective To retrospectively evaluate the relevant factors for hepatitis B virus-associated glomerulonephritis (HBV-GN).Methods A total of 86 patients with pathologyproven HBV-GN and 135 HBV carriers with non-HBV-GN were included in this retrospective casecontrol study.Logistic regression analysis was used to detect the relevant factors for HBV-GN.Results On univariate analysis,the factors associated with HBV-GN were as follows: male (OR 2.79,95%CI 1.48-5.25,P=0.001),HBeAg positivity (OR 2.60,95%CI 1.49-4.53,P=0.001),HBV replication (OR 3.63,95%CI 1.80-7.33,P＜0.01),liver cirrhosis (OR 4.58,95%CI 1.41-14.91,P=0.011),and elevated alanine aminotransferase (ALT) (OR 2.53,95%CI 1.42-4.51,P=0.002).On multivariate analysis,the associations remained significant for male (OR 2.21,95%CI 1.12-4.33,P=0.022),HBV replication (OR 2.77,95%CI 1.28-5.97,P=0.01),liver cirrhosis (0R 4.55,95%CI 1.29-16.10,P=0.019) and elevated ALT (OR 1.96,95%CI 1.04-3.69,P=0.037).Compared with HBV-associated IgA nephritis (HBV-IgAN) in multivariate model,HBV-associated membranous nephropathy (HBV-MN) or membranoproliferative glomerulonephritis (HBV-MPGN) was significantly associated with male (OR 6.51,95%CI 1.76-24.11,P=0.005) and HBV replication (OR 7.22,95%CI 1.68-30.97,P=0.008).Conclusions Male,HBV replication,liver cirrhosis and elevated ALT may be predictive factors for HBV-GN.Compared with HBV-IgAN,HBV-MN or HBV-MPGN is significantly associated with male and HBV replication.

Objective To explore the effect and mechanism of Baicalein in the treatment of hyperuricemia.Methods The mouse model of hyperuricemia was established by yeast extract combined with potassium oxazinate.The effect and potential mechanism of Baicalein in the treatment of hyperuricemia were studied by biochemical indexes,pathological changes,non-target metabonomics and network pharmacology.Results Baicalein could reduce the contents of serum uric acid,creatinine and blood urea nitrogen,reduce the inflammatory injury of renal tissue,up-regulate the expression level of uric acid excretion protein and down-regulate the expression level of uric acid reabsorption protein.Nine disease-related targets such as BCL2,SIRT1 and XDH were screened by network pharmacology.Six key metabolic pathways including nicotinic acid and nicotinamide metabolism,caffeine metabolism and purine metabolism were screened by metabonomics analysis.Conclusions Baicalein can treat hyperuricemia and reduce renal injury,and its mechanism may be related to the metabolic pathways of nicotinic acid and nicotinamide regulated by SIRT1 and quinolinate.

Objective To evaluate the efficacy of laparoscopic radical resection for rectal cancer by collecting and analyzing long-term outcomes of patients and to investigate prognostic factors of overall survival and disease free survival.Methods The clinicopathological data of 235 patients who underwent laparoscopic radical resection for rectal cancer from Jan 2007 to Dec 2010 were retrospectively analyzed.COX proportional hazards regression model was used to determine the risk factors for overall survival and disease free survival.Results A total of 235 patients were included in this analysis.Local recurrence rate were 8.1％ at 3 years and 9.8％ at 5 years.Overall and disease free survival were 85.2％ and 75.1％ at 3 years,77.1％ and 69.6％ at 5 years,respectively.Factors found to significantly and independently predict a poor overall and disease free survival were laparoscopic Hartmann,postoperative complications,stage Ⅲ tumor and ulcerative type tumor.Neural invasion was also an adverse prognostic factor of overall survival.Conclusions Laparoscopic Hartmann,postoperative complications,stage Ⅲ tumor and ulcerative type tumor were independently associated with overall and disease free survival.In addition to this,neural invasion was also an adverse prognostic factor of overall survival.

Objective To evaluate the diagnostic value of T cells spot test of tuberculosis infection (T-SPOT .TB) on blood and cerebrospinal fluid for tuberculous meningitis (TBM ).Methods One hundred and fifteen adult patients with suspected tuberculous meningitis were retrospectively enrolled from March 2013 to March 2017 in Huashan Hospital affiliated to Fudan University .Among them ,30 were diagnosed with TBM (7 definite ,19 highly probable and 4 possible) ,37 with other infectious meningitis and 29 with non-infectious meningitis .The diagnostic sensitivity ,specificity ,positive predictive values (PPV) and negative predictive values (NPV) of T-SPOT .TB on peripheral mononuclear cells (PBMC) and cerebrospinal fluid mononuclear cells (CSF-MC ) were analyzed using Fisher exact test , and the diagnostic performance was evaluated by using receiver operating characteristic (ROC ) curve and area under the curve (AUC).Results When including the 30 TBM cases and 66 non-TBM cases into analysis , the sensitivities and specificities , PPV and NPV of PBMC and CSF-MC for diagnosing TBM were as follows :93 .1% and 66 .7%,77% and 87 .7%,65 .9% and 71 .4%,95 .9% and 85 .1%,respectively . When including the 30 TBM and 37 other infectious meningitis into analysis , the sensitivities and specificities ,PPV and NPV of the PBMC and CSF-MC for diagnosing TBM were as follows :93 .1% and 66.7%,68 .6% and 86 .5%,71.1% and 80 .0%,92 .3% and 76 .2%,respectively .By ROC curve analysis ,the AUC of blood and CSF were 0 .882 (95% CI :0 .795－0 .969) and 0 .814 (95% CI :0 .704－0 .925) ,respectively .Using a cut-off value of 32 spot forming cells (SFC) per million CSF-MC for T-SPOT .TB on CSF-MC showed a sensitivity of 66 .7%,a specificity of 91 .9%,PPV of 87 .0% and NPV of 77 .3% .The positive likelihood ratio and negative likelihood ratio were 8 .22 and 0 .363 respectively . Conclusions T-SPOT .TB on CSF-MC has a role in diagnosing TBM .And 32 SFC per million CSF-MC might be the optimal cut-off value to differentiate TBM and non-TBM .

Objective: To analyze the clinical data of the elderly peritoneal dialysis (PD) patients in Peking Union Medical College Hospital (PUMCH), and to find the risk factors for the long-term survival. Methods: Baseline data and the outcome of maintenance PD patients from 1996-03 to 2015-09-30 were collected for a retrospective cohort study. Patients were divided into the non-elderly group (<65 years old), the 65-79 years old group and the ≥80 years old group, and were follow to 2016-09-30. The survival rate was calculated by Kaplan-Meier method and the risk factors of outcome were analyzed by the Cox's regression model. Results: Among 577 PD patients, about 243(42.1%) were elderly patients, including 207 patients aged between 65 and 79 years (35.9%) and 36 patients aged 80 or more (6.2%). The most common primary disease causing PD was diabetic nephropathy (DN) for both elderly and non-elderly patients. The 1-year, 3-year, 5-year survival rate of patients aged between 65 and 79 years were 87.0%, 61.9%, 32.4% respectively, and 72.5%, 48.5%, 27.3% for the ≥80 years old group. The dominating reasons of death were cardiovascular events and infection. There was no difference of technical survival rates among three groups, and the most common reason for technical failure was peritonitis. For elderly patients, diabetes (HR=2.193, 95% CI 1.445-3.328, P<0.001) and lower baseline serum albumin (HR=0.968, 95%CI 0.940-0.996, P=0.026) were independent risk factors for death. However, for non-elderly patients, diabetes (HR=3.746, 95%CI 2.149-6.529, P<0.001) was the only independent risk factor for death. Conclusions Cardiovascular diseases and infection are the main reasons for death among the elderly PD patients in PUMCH. Diabetes and lower baseline serum albumin may predict the mortality of elderly PD patients independently. Better management of nutrition might improve survival in elderly PD patients.