BACKGROUND:Establishing the animal model of membranous nephropathy is of importance to figure out the pathogenesis of membranous nephropathy.
OBJECTIVE:To investigate the expression of nephrin and podocin in the model of membrane nephropathy in rats, and to investigate their relationships with the pathogenesis of membranous nephropathy.
METHODS:A total of 40 Sprague-Dawley rats were equivalently randomized into model and control groups. Rats in the model group were in premunity by given subcutaneous and multi-point injection of 1 mg cationic bovine serum albumin firstly dissolved in 0.5 mL normal saline and then ful y emulsified with the equal incomplete Freund’s adjuvant for 1 week, and 16 mg/kg cationic bovine serum albumin was injected via vein tails, once every other day for 4 weeks. The same volume of normal saline was injected into the controls. The mRNA expressions of nephrin and podocin in renal tissues were detected using real-time PCR, and biochemical indicators and morphological observation were measured at 2 and 4 weeks after modeling.
RESULTS AND CONCLUSION:(1) In the model group, the total amount of urine and serum albumin levels were significantly decreased accompanying with overt proteinuria, and the serum creatinine, urea nitrogen, cholesterol and triglyceride levels were significantly increased al in a time-independent manner compared with the control group (P<0.01), and the difference was significant (P<0.05). (2) The pathological examination showed that rats in the model group had different degrees of renal tubular dilatation, glomerular basement membrane thickening, mesangial cel s and stromal hyperplasia, which was typical of membranous nephritis. (3) Moreover, the mRNA expressions of podocin and nephrin in the model group were lower than those in the control group. (4) In conclusion, the decreased expressions of podocin and nephfin may disturb the integrity of the slit membrane of podocytes giving rise to the damage of glomerular filtration barrier, and proteinuria appears in final.

Objective To explore the changes of MR diffusion imaging (DWI) appearance in newborn rats with hypoxic‐ischemic brain damage(HIBD) ,and its relationship with the changes of Triphenyl tetrazolium chloride (TTC) staining .Methods Using liga‐tion of the left carotid artery method to establish three different degrees of HIBD animal models ;DWI was performed at each time point(6-12 h ,12-24 h ,3 d ,7 d);Fresh brain tissue taken from another model groups of newborn rats in 12 h ,24 h ,3 d ,7 d were staining in TTC ,then we observed its relationship with DWI .Results The lesion location of three model groups mainly distributed in the left side of cortex and subcortical region ,with prolonged hypoxia time ,hippocampus ,lateral side white matter ,thalamus were also have varying degrees of involvement .The right side of the cortex and subcortical in some cases involved .TTC staining showed posi‐tive results in 3 d ,its loss stained area were consistent with DWI abnormal signal area .Conclusion DWI can be evaluation of HIBD lesions early .The early lesions of HIBD mainly distributed in the left side of cortex and subcortical region .

Objective To explore the application of diffusion tensor imaging(DTI) in neonates with hypoxic ischemic encephalopathy(HIE).Methods 37 cases of full-term neonates diagnosed as HIE by pediatricians and 12 normal neonates were selected.All subjects underwent MRI and DTI at 3.0 Tesla.The correlation of FA value in posterior limb of internal capsule and neonatal NBNA score was examined.Results The FA values in posterior limb of internal capsule, splenium had significant difference between the HIE group (mild group, moderate group, severe group) and control group (P<0.05);in central white matter of the frontal lobe, thalamus had significant difference between HIE severe group and control group (P<0.05);in nucleus of the bean had no statistic difference between the HIE group (mild group, moderate group,severe group) and control group (P>0.05).The FA in posterior limb of internal capsule and NBNA score(12-14 d;26-28 d)showed significant correlation (r=0.96,P<0.05),as well as the sequelae shape (r=0.87,P<0.05).Conclusion DTI is promising for early diagnosis of anisotropic index changes to evaluate HIE brain damage and prognosis.

Objective To investigate the comparison of clinical effects between two models of alimentary reconstruction after total gastrectomy in the elderly patients with gastric cancer.Methods 70 patients receiving alimentary tract reconstructions of Orr-type Roux-en-Y reconstruction (Orrtype,40 cases) and modified Brawn Ⅰ (30 cases) from January 2007 to December 2012 were retrospectively analyzed.The operative time,amount of bleeding,early postoperative complications and mortality,food intake,nutritional status and alimentary tract function were compared at 6 months after surgery.Results There were no significant differences between the two methods in the operative time [(198.8±14.0)min vs 233.5±30.7)min,t=-l.697,P＞0.05)],amount of bleeding [(420 ± 43) ml vs (340 ± 25) ml t =-1.956,P ＞ 0.05],and early postoperative complications [(17.5％ vs 16.7％),x2 =0.008,P＞0.05].However,times of liquid diet intake and semi liquid diet intake of the Orr-type reconstruction method was comparatively shorter than that of the modified Brawn Ⅰ [(4.8±2.1) d vs (7.6±2.4) d,and (9.5±3.6) d vs (11.5±3.7) d,t=-5.192,P＜0.05)].Nutritional status (weight,hemoglobin,total serum proteins and serum albumin) at 6 months after operation showed no significant differences between the two methods.But constituent ratio of Visick scores Ⅰ-Ⅱ of Orr-type was bigger than that of modified Brawn Ⅰ (86.7％ vs 62.5％,x2 =5.063,P＞0.05).Conclusions Orr-type Roux-en-Y reconstruction method can avoid reflux esophagitis,and the procedure is more simple than the modified Brawn Ⅰ method.Therefore,Orr-type Roux-en-Y reconstruction should be recommended as an adoptable method of digestive reconstruction after total gastrectomy for gastric cancer in the elderly patients.

Objective To evaluate the surgical modalities and prognosis in elderly patients with gastric cancer.Methods Sixty three gastric cancer patients aged ≥≥ 75 years underwent radical surgery,with D0 + D1 resection in 32 cases and D2 resection in 31 cases,in Beijing Hospital from January 2005 to December 2009.Results More lymph nodes were dissected in D2 group than those in D0 + D1 group (27.42 ± 12.75 vs.14.59 ± 12.11,t =-4.095,P ＜ 0.05).There was no significant difference in postoperative complication rate [25.81％ (8/31) vs.31.25％ (10/32),x2 =0.095,P ＞ 0.05] and perioperative death rate [3.23％ (1/31) vs.6.25％ (2/32),x2 =0.324,P ＞ 0.05] between two groups.The 5-year survival rate was higher in D2 group than that in D0 ± D1 group (47.3％ vs.21.4％,x2 =6.346,P ＜ 0.05).The 5-year survival rate was higher in POSSUM score (PS) ＜ 20 group than that in PS≥20 group (46.3％ vs.11.1％,x2 =5.492,P ＜0.05).In PS≥20 group,postoperative complication rate did not increase after D2 radical resection(1/8 vs.3/10,x2 =0.824,P ＞ 0.05).Univariate analysis showed that PS ＜ 20 was associated with the prognosis of patients (x2 =5.492,P ＜ 0.05).Cox proportional hazards model showed that lymph node metastasis (OR=4.103,95％ CI:1.790-9.405,P ＜ 0.05) and D2 radical gastrectomy(OR =0.313,95％ CI:0.158-0.620,P ＜ 0.05) were the independent factors associated with the prognosis of patients aged ≥ 75 years with gastric cancer (all P ＜ 0.05).Conclusion Standardized D2 lymph node dissection is beneficial for gastric cancer patients aged ≥75 years.PS ＜ 20,lymph node metastasis and D2 radical gastrectomy are associated with the prognosis of patients.

Objective To observe the clinic efficacy of open transforaminal lumbar interbody fusion (TLIF) compared with minimally invasive operation in treating lumbar spinal stenosis and instability among obese and non‐obese patients .Methods A ret‐rospective analysis was performed in these cases of mono‐segmental lumbar spinal stenosis and instability between January 2011 and January 2013 .Perioperative index ,clinical efficacy ,and imaging results were observed and compared between different groups .Re‐sults Thirty‐four obese cases and 105 non‐obese cases were divided into two groups ,including conventional posterior open TLIF and minimally invasive TLIF operation ,to compare the results .Perioperative indexes of obese patients were more than non‐obese patients undergone open TLIF operation way and there was significant difference(P<0 .01);while by minimally invasiveTLIF oper‐ation way ,comparing the results of the surgical blood loss and incision size between obese and non‐obese group ,there was signifi‐cant difference(P<0 .05) .However ,there was not significant difference in the operative time between two groups(P>0 .05) .No cases of slippage or breakage of implants were found among all these patients after 6 months of follow up .Postoperative VAS and ODI among these four groups were better than before(P<0 .01) .Undergoing open postoperative VAS in obese group and in non‐obese group ,there was significant difference(P< 0 .05);undergoing minimally invasive postoperative ODI in obese group and in non‐obese group ,there was significant difference(P< 0 .05) .But ,undergoing open postoperative ODI in obese group and in non‐obese group ,there was not significant difference(P>0 .05);undergoing minimally invasive postoperative VAS in obese group and in non‐obese group ,there was not significant difference(P>0 .05) .Conclusion Therefore ,obese may be risk factor in treating lumbar spinal stenosis and instability .

Objective To study the clinical value of using a four-point method (FP) in measuring joint angles.Method Methods Twenty-five subjects without knee dysfunction were studied.Every knee joint was tested in extension,below 90° of flexion and above 90° of flexion with the traditional three-point method (TP) and the new four-point method.X-rays were taken as a gold standard measurement.The reliability and validity were analyzed using Bland-Altman plots,which can yield 95% limits of agreement (LOA).Results (1) The joint axis centers moved with changes in joint angle.(2) The 95% LOA of inter-tester reliability was (-5.0,2.7) with the FP and (-6.5,4.4) with the TP.The 95% LOAs of intra-tester reliability were (-3.6,4.0) for tester A and (-4.1,4.8) for tester B with the FP,while (-5.0,6.4) for tester A and (-6.1,5.8) for tester B with the TP.(3) The 95% LOAs in the three positions were (3.5,2.6) in extension,(-3.3,3.6) in mild flexion and (-1.9,5.7) in greater flexion with the FP,and ( 6.8,1.1),(-4.0,7.1) and (0,8.5) with the TP.Conclusions The FP gives better reliability and validity than the TP in joint angle measurement.

AIM:To explore the influences of semaphorin 3A (Sema 3A) on hydrogen peroxide (H2O2)-induced injury in human umbilical vein endothelial cells (HUVECs).METHODS:Sema 3A over-expression vectors were constructed and transfected into the HUVECs by Lipofectamine 2000, and the over-expression effect was verified by qPCR and Western blot.The HUVECs in different groups were treated with or without 200 μmol/L H2O2 for 4 h.The levels of inflammatory cytokines were measured by qPCR.The levels of lactic dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by corresponding colorimetry.The cell viability was measured by MTT assay.The cell apoptosis was analyzed by flow cytometry.The levels of apoptosis-related proteins cleaved caspase-3 and Bcl-2 were determined by Western blot.RESULTS:H2O2 induced inflammatory cytokine secretion, increased the levels of LDH and MDA, decreased SOD activity and cell viability, and increased cell apoptosis in the HUVECs.Over-expression of Sema 3A enhanced the above processes.No injury effect of Sema 3A over-expression on HUVECs without H2O2 treatment was observed, indicating that the injury effects of Sema 3A on HUVECs depended on H2O2.CONCLUSION:Sema 3A markedly enhances H2O2-induced injury in the HUVECs, which depends on H2O2.Sema 3A may promote oxidative stress-caused endothelial cell injury.

OBJECTIVETo investigate the predictive value of preoperative Glasgow prognostic score (GPS) for the postoperative complications and survival in patients with colorectal cancer (CRC) undergoing laparoscopic radical resection.

METHODSThis retrospective study was conducted in the Beijing Hospital between January 2009 and January 2012. A total of 228 patients with primary CRC undergoing laparoscopic radical resection were analyzed. The GPS was constructed based on routine preoperative blood tests of C-reactive protein and serum albumin. The patients were classified into three groups according to GPS (GPS 0, 1, 2 groups). Survival curves were described by the Kaplan-Meier method and compared by the Log-rank test. The univariate and multivariate analyses were performed with the Cox proportional hazard model to identify the influence of GPS on prognosis in patients with CRC undergoing laparoscopic radical resection.

RESULTSPreoperative CRP level was increased in 48 cases (21.1%), and preoperative serum albumin level was decreased in 104 cases (45.6%) in the whole group. These 228 patients were classified into 99, 105 and 24 patients in GPS 0, 1, 2 group respectively. GPS was significantly associated with age, preoperative body mass index (BMI), carcinoembryonic antigen (CEA), CA19-9, tumor location, tumor differentiation and TNM stage (all P<0.05). Postoperative complication rates of GPS 0, 1, 2 group were 6.1%, 14.3% and 70.8% respectively (χ=59.147, P=0.000). Serious postoperative complication rates were 3.0%, 6.7% and 58.3% respectively (χ=65.807, P=0.000). Univariate and multivariate analyses revealed that GPS was an independent risk factor of postoperative complications(HR=21.611, 95%CI: 5.936-78.681, P=0.000) and severe complications (HR=35.833, 95%CI: 7.364-174.355, P = 0.000). The 5-year survival rate was 50% and the average total survival time was 58.2 (95% CI: 54.6-61.7) months in the whole group. The median overall survival time in GPS 0, 1, 2 group was 74.6(95%CI: 70.4-78.7) months, 49.8(95%CI: 45.2-54.4) months and 27.8 (95%CI: 21.8-33.8) months respectively(χ=98.425, P=0.000). The median disease-free survival time was 73.9(95%CI: 69.2-78.7) months, 47.4 (95% CI: 41.6-53.1) months and 19.9 (95%CI: 14.8-25.0) months respectively (χ=91.305, P=0.000). GPS was an independent risk factor of disease-free survival (HR=4.840, 95%CI: 2.413-9.709, P=0.000) and overall survival (HR=6.267, 95%CI: 3.073-12.784, P=0.000).

CONCLUSIONSGPS can be used as an effective predictor of the prognosis for patients with CRC undergoing laparoscopic radical surgery. Higher GPS suggests more postoperative complications and worse prognosis.

Adult ; Aged ; Biomarkers, Tumor ; analysis ; C-Reactive Protein ; analysis ; CA-19-9 Antigen ; analysis ; Carcinoembryonic Antigen ; analysis ; Colorectal Neoplasms ; blood ; surgery ; Disease-Free Survival ; Female ; Humans ; Laparoscopy ; Male ; Middle Aged ; Multivariate Analysis ; Postoperative Complications ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Risk Factors ; Serum Albumin ; Survival Rate

Aim Toconstructthreeplasmidsincluding Smad3 WT,Smad3 EPSM and Smad3 3S-A stable transfection in HepG2 cell lines to investigate phospho-domains of Snad3(pSmad3C or pSmad3L),their pro-tein expression and roles in HepG2 cell proliferation, apoptosisandcellcycle.Methods Threeplasmidsin-cluding Smad3 WT (Carry the wild Smad3 gene ), Smad3 EPSM(Carry the mutated phosphorylation site in linker region of Smad3 gene)and Smad3 3S-A(Car-ry the mutated phosphorylation site in C-terminal of Smad3 gene)were respectively transfected into HepG2 cells by using a liposome transfection reagent.Verifi-cation of positive cells was done by screening with G418 via co-culture.Transfection efficiency was deter-mined by Western blot.Cell proliferation was induced by exogenous TGF-β1 in the respective stably transfect-ed HepG2 cell lines.Cell proliferation was monitored by MTT.Cell cycle and apoptosis were determined by flowcytometry(FCM).Results Therewaselevated protein expression of the respective phospho-domain sites in the stably transfected HepG2 cells for Smad3 WT(C-terminus and Linker),Smad3 EPSM(C-termi-nus)and Smad3 3S-A(Linker),which indicated suc-cessful stable transfection of HepG2 cell lines.The re-sults from MTT experiment showed that TGF-β1 could induce proliferation of HepG2 cells with or without the transfection of Smad3 WT,Smad3 EPSM and Smad3 3S-A plasmids,meanwhile transfected Smad3 EPSM plasmids could significantly inhibit proliferation of HepG2 cells induced by TGF-β1 , and transfected Smad3 3S-A plasmids accelerate proliferation of HepG2 cells induced by TGF-β1 .Cell cycle analysis showed that the G0/G1 phase of HepG2 cells with stable trans-fection of Smad3 EPSM plasmid increased compared with HepG2 cells with or without stable transfection of Smad3 WT plasmid,meanwhile the G2/M phase of HepG2 cells with stable transfection of Smad3 3 S-A plasmid increased.Compared with Smad3 WT trans-fected cells, apoptosis in Smad3 EPSM transfected cells was markedly increased,while that of Smad3 3S-Atransfectedcellsdecreased.Conclusions Thethree plasmids of Smad3 WT,Smad3 EPSM and Smad3 3S-A stably transfected HepG2 cell lines have been suc-cessively constructed.The construction of three plas-mids transfected HepG2 cell lines provides the research foundation for studying medical as well as possible reg-ulatory mechanism of pSmad3 C/pSmad3 L.