Objective To investigate whether the Notch-1 signaling pathway is involved in the acquisition of the epithelial-mesenchymal transition (EMT) phenotype of gefitinib-acquired resistant lung cancer cells.Methods The PC9 cell line (harboring EGFR exon 19 deletion) and PC9/AB2 cells (gefitinibacquired resistant PC9 cells) were used.siRNA targeting Notch-1 eukaryotic expression vector (siNotch-1) was constructed and PC9/AB2 cells were transfected with siNotch-1.The protein expression of EGFR,Akt,Erk,Notch receptors and ligands,TGF-β receptors,E-cadherin,Vimentin,and Snail were detected by Western blot assay.DNA sequencing and fluorescence in situ hybridization (FISH) analysis were processed to detect mutation of EGFR exon 20 and MET amplification,respectively.For cytotoxicity assay,cell viability was assessed by Cell Counting Kit 8.Results Gefitinib resistant cell line PC9/AB2 had no evidence of MET amplification or EGFR T790M mutation.The expression of Notch-1 was upregulated in gefitinib resistant PC9/AB2 cells compared with that in gefitinib-sensitive PC9 cells.There were no significant protein expression differences of other Notch receptors,Notch ligands or TGF-β receptors between both paired cell lines.Western blot results showed that protein expression of E-cadherin was greatly reduced in PC9/AB2 cells,while elevated levels of Vimentin and Snail were observed.A significant reduction of the expression of Snail and Vimentin in Notch-1 siRNA transfected PC9/AB2 cells with increased E-cadherin expression was found by Western blot assay.PC9/AB2 cells displayed a round like cell morphology after Notch-1 siRNA transfection.Silence of Notch-1 decreased colony-formation ability but enhanced sensitivity of gefitinib on PC9/AB2 cells.Conclusion Notch-1 might play a novel role in acquired resistance to gefitinib,which could be reversed by inhibiting Notch-l.

ObjectiveTo explore the feasibility and outcomes of natural orifice transanal laparoscopic Soave procedure for Hirschsprung's disease and allied disorders (HAD).MethodsFrom March 2010 to December 2011,31 cases (at the age from 3 mos to 6 yrs) with Hirschsprung's disease or allied disorders (5 cases)underwent laparoscopic-assisted Soave pull-through procedure at two tertiary hospitals.We modified this technique by mobilizing the left hemicolon or whole colon via rectal muscular sleeve approach under transanal or transumbilical laparoscopic vision,then endorectal pull-through to completearectosigmoidectomyorsubtotalcolectomy. ResultsAllprocedureswerecompleted successfully.A rectosigmoidectomy was performed in 16 cases with classic HD and subtotal colectomy in 15 cases with extended HD and HAD.The average operative time was ( 117 ± 13) min.The length of the resected segment was 35 -80 cm,and the estimated blood loss was 5 -20 ml. One infant developed postoperative intestinal obstruction that required open exploration.Follow-up of one to 20 mos found no stoma stenosis or constipation recurrence. Enterocolitis developed in 1patient.ConclusionsTransanal or transumbilical laparoscopic-assisted Soave pull-through surgery is safe,effective and with a benefit of much less invasion and almost invisible scars.

AIM: To explore the role of TGF-?_1 and signaling transduction molecule, Smad4, in the development of glomerulosclerosis. METHODS: Expression levels of TGF-?_1, Smad4, collagen Ⅰ proteins were evaluated by immunohistochemistry in renal biopsies from 38 cases with a spectrum of glomerulonephritis, and compared with 20 normal kidney tissue with image analysis system. After stimulation with TGF-?_1, expressions of endogenous Smad4 and collagen Ⅰ mRNA and proteins and its modulation by TGF?_1 were evaluated by RT-PCR and Western blotting analyses in cultured human mesangial cells. RESULTS: All types of proliferative and sclerotized glomerulonephritis showed an increased expression of TGF-?_1, Smad4 and collagen Ⅰ ompared with the 20 normal kidney tissue in glomerular (P

0.05). CONCLUSION: Serum Cys C is significantly increased in ARF and correlated well with the severity of ARF. Serum Cys C can be one of the detectable markers of ARF, but it is independent of the mortality and does not predict the prognosis of these patients.

Objective To explore the risk factors of hypertension in patients with IgA nephropathy in South China. Methods The clinical and renal pathological data of 280 primary IgA nephropathy patients diagnosed by biopsy were analyzed to extinguish the risk factors of hypertension. Results A total of 96 patients were suffered with hypertension (34.3%). A single-variable analysis showed that the age (≥40 years), body weight (≥60 kg), absence of macrohematuria, duration of disease (≥60 months), blood urea nitrogen≥8 mmol/L, serum creatinine (≥133 μmol/L), hyperuricaemia, degree of 24 h-proteinuria (≥1.5 g), segmental glomerular lesions (≥25% ), globe glomerular sclerosis (≥10%), tubular atrophy (≥25%), interstitial fibrosis (≥25%), interstitial inflammation (≥25% ) and arteriole hypertrophy (≥10% ) were all risk factors related to hypertension; multivariate logistic regression analysis showed that serum creatinine, age, arteriole hypertrophy, body weight and 24 h-proteinuria were the independent risk factors. Conclusion Many factors were related the hypertension in patients with IgA nephropathy, while serum creatinine, age, arteriole hypertrophy, body weight and 24 h-proteinuria were the independent risk factors of hypertension.

OBJECTIVETo establish a prognostic model for predicting extracorporeal circulation clotting in patients with continuous renal replacement therapy (CRRT).

METHODS425 patients with CRRT were involved in the study. We built a predictive risk model of extracorporeal blood clotting with the 302 participants, and 103 participants were used to validate the model. The primary endpoint of CRRT was extracorporeal circulation pipe blockage.

RESULTSWe used a score of 0-5 point evaluating system to predict the risk of 24 hours CRRT integral model of cardiopulmonary bypass clogging. The area under the CRRT predictive model of cardiopulmonary bypass clogging integral system ROC curve was 0.790 (95% CI 0.719-0.826) (P<0.001). The evaluating system can determine the blockage of 24 hours CRRT extracorporeal circulation. The results showed that CRRT extracorporeal plugging prediction fitted the integral model and could predict the chance of plugging. The actual plugging rate showed no significant difference from the predicted rate (R² = 0.301, P=0.232). The cardiopulmonary pipe survival time between the 3 groups(low risk, intermediate risk, and high risk) showed a significant difference (P<0.05).

CONCLUSIONWe established a continuity extracorporeal blood purification plugging risk score model, to predict plugging risks during CRRT treatment.

Blood Coagulation ; Extracorporeal Circulation ; Humans ; Models, Theoretical ; Prognosis ; ROC Curve ; Renal Replacement Therapy ; Risk Assessment

Objective:To study the treatment efficacy and safety in using needle-grasper assisted single-port laparoscopic hepaticojejunostomy to treat choledochal cysts in children.Methods:The data of 41 patients with choledochal cysts treated at the Department of Pediatric Surgery, the First Affiliated Hospital of Xiamen University and the Second Hospital of Hebei Medical University from January 2018 to December 2021 were reviewed. There were 8 males and 33 females, aged (2.5±1.9) years. These patients were divided into the needle-grasper assisted single-port laparoscopic hepaticojejunostomy group (needle-grasper group, n=21) and the single-port laparoscopic hepaticojejunostomy group (control group, n=20). Operation time, intestinal function recovery time, gastric tube retention time, abdominal drain indwelling time, postoperative hospital stay, and perioperative complications were compared between the two groups. Results:All 21 children in the needle-grasper group underwent successful surgery without any need to convert to conventional laparoscopic or open surgery. The operation time (156.4±21.2) min was significantly shorter than the control group (218.3±28.6) min ( t=2.95, P=0.017). There were no significant differences in intestinal function recovery time, gastric tube retention time, abdominal drain indwelling time, postoperative hospital stay and perioperative complications between the two groups (all P>0.05). Parents were very satisfied with the cosmetic effect of the invisible scar after surgery. Conclusion:Needle-grasper assisted single-port laparoscopic hepaticojejunostomy was safe, reliable and the operation time was shorter than using a single-port to achieve minimally invasive and scarless surgery.

OBJECTIVETo observe the effect of amiloride on the proteinuria of the 5/6 nephrectomy rats.

METHODSTo establish the 5/6 nephrectomy rats model and divide the experiment into 3 groups, sham operated group(Sham), 5/6 nephrectomy model group(NTX) and 5/6 nephrectomy with amiloride-treated group (NTX+amiloride, n=15). The concentration of protein and mRNA of uPAR and the change of podocytes motility were detected by coomassiebluestaining, immunofluorence method and real-time PCR.

RESULTSAt second week, compared with Control group, the 24 h urine protein of NTX group was significantly increased (47.50 ± 28.05 mg vs 14.28 ± 3.8 mg, P = 0.023). There was no statistical significance in 24-hour urine protein between NTX+amiloride group and NTX group (51.56 ± 21.03 mg vs 47.50 ± 28.05 mg, P = 0.748). The same situation was also observed at the time point of 12 week, comparing with NTX group, 24-hour urine protein decreased in Sham group (188.31 ± 29.82 mg vs 21.32 ± 8.59 mg, P = 0.000) and NTX+amiloride group (188.31 ± 29.82 mg vs 121.37 ± 31.14 mg, P=0.000), with statistical significance when comparing with Sham group, the expression of uPAR mRNA in NTX group was significantly increased (9.74 ± 1.44 vs 1.01 ± 0.13, P = 0.000). In contrast, the expression of uPAR mRNA in NTX rats treated with amiloride was significantly lower than in NTX group (9.74 ± 1.44 vs 5.01 ± 1.36, P = 0.000).

CONCLUSIONAmiloride can reduce the proteinuria of the 5/6 nephrectomy rats model of transient proteinuria by inhibiting the induction of uPAR expression.

Amiloride ; pharmacology ; Animals ; Cell Movement ; Disease Models, Animal ; Nephrectomy ; Podocytes ; drug effects ; metabolism ; Proteinuria ; drug therapy ; Rats ; Real-Time Polymerase Chain Reaction ; Receptors, Urokinase Plasminogen Activator ; metabolism

PURPOSE: Adenoid cystic carcinoma (ACC) of the trachea and bronchus is a rare tumor. Although MYB-NFIB oncogene fusion and Notch1 mutation have been identified in ACC, little is known about the expression and clinical significance of Notch1 and its target gene fatty acid binding protein 7 (FABP7) in tracheobronchial ACC. MATERIALS AND METHODS: Primary tracheobronchial ACC that were resected between 1998 and 2014 were identified through the pathology and oncology database from five thoracic oncology centers in China. A tissue array was constructed from the patients’ samples and the expressions of Notch1 and FABP7 were evaluated by immunohistochemistry. The association between the expression of both markers and survival was determined. RESULTS: Overexpression of Notch1 and FABP7, detected in 37.8% and 38.3% of 368 patients with tracheobronchial ACC, respectively, was an independent prognostic indicator for recurrencefree survival (RFS) by multivariable Cox proportional hazard model (p=0.032 and p=0.048, respectively). Overexpression of Notch1, but not of FABP7, predicted overall survival (OS) (p=0.018). When categorized into four groups according to coexpression of Notch1 and FABP7, patients with overexpression of both Notch1 and FABP7 belonged to the group with the shortest RFS and OS (p=0.01 and p=0.048, respectively). CONCLUSION: Expression of Notch1 and FABP7, and coexpression of Notch1 and FABP7, is strongly associated with poor survival in resected tracheobronchial ACC. These data are consistent with the hypothesis that poor differentiation of tracheobronchial ACC correlates with the activation of Notch signaling.
Adenoids* ; Bronchi ; Carcinoma, Adenoid Cystic* ; Carrier Proteins* ; China ; Humans ; Immunohistochemistry ; Oncogene Fusion ; Pathology ; Prognosis ; Proportional Hazards Models ; Retrospective Studies* ; Trachea

OBJECTIVETo determine whether hyperglycemia activates NFAT2 in cultured podocytes to cause podocyte apoptosis and explore the role of NFAT2 in high glucose-induced podocyte apoptosis.

METHODSImmortalized mouse podocytes were cultured in the presence of normal (5.3 mmol/L) or high glucose (10, 20, 30, and 40 mmol/L) or pretreated with 11R-vivit (100 nmol/L) or cyclosporine A (500 nmol/L) before exposure to 20 mmol/L glucose for different durations (0.5-48 h). The activation of NFAT2 in the podocytes was detected using Western blotting and immunofluorescence assay. The role of NFAT2 in hyperglycemia-induced podocyte apoptosis was explored by observing the inhibition of NFAT2 activation by 11R-vivit using flow cytometry. Intracellular Ca was monitored in high glucose-treated podocytes using Fluo-3/AM. The mRNA and protein expressions of the apoptosis gene Bax were detected using real time-qPCR and Western blotting.

RESULTSExposure to high glucose in the medium time- and dose-dependently activated NFAT2 in cultured podocytes. Pretreatment with cyclosporine A or 11R- VIVIT completely blocked nuclear accumulation of NFAT2. Treatment with 11R- vivit also inhibited high glucoseinduced apoptosis in cultured podocytes. Exposure to high glucose obviously increased [Ca]I in the podocytes to cause activation of calcineurin and the subsequent increment of nuclear accumulation of NFAT2 and Bax expression.

CONCLUSIONSHigh glucose-induced apoptosis in podocytes is mediated by calcineurin/NFAT2/Bax signaling pathway, which may serve as a potential target for therapeutic intervention.