Objective To investigate the possible injury mechanism of human kidney proximal tubular epithelial cell-2 (HK-2) induced by aristolochic acid (AA). Methods Cultured HK-2 cells were divided into 4 groups:normal control,treated by AA at the concentration of 30,60 and 120 ?mol/L for 48 h respectively. The morphological changes were observed by inverted phase contract microscopy. The cell viability was measured by the Cell Counting Kit-8 (CCK8) assay. Apoptotic cells were identified by flow cytometry. Expression of active Caspase-3 was measured by Western blot analysis. Automatic biochemical analyzer was used to detect the contents of LDH and ?-N-Acetylglucosaminidase (NAG) in the supernatant. The expression of E-cadherin and a-SMA was detected with laser scanning confocal microscope (LSCM). Enzyme linked immunosorbent assay (ELISA) was used to measure the levels of TGF-?1 and collagen Ⅲ in the supernatant quantitatively. Results AA inhibited HK-2 cells proliferation,induced cell apoptosis and activated Caspase-3 expression,and increased the LDH and NAG levels. All of these were in a concentration-dependent manner. AA at the concentration of 60 ?mol/L inhibited E-cadherin expression,increased ?-SMA expression and TGF-?1 and collagen Ⅲ secretion. Conclusion AA inhibits cell proliferation,induces apoptosis and epithelial-mesenchymal transition (EMT) in HK-2 cells. AA at relatively low concentration (≤60 ?mol/L) mainly induces EMT in HK-2 cells,while,that at high concentration (≥120 ?mol/L) causes apoptosis and cytotoxicity.

Objective To observe the activation of pancreatic stellate cells (PSC) during the formation of pancreatic fibrosis induced by the pancreatic injection of trinitrobenzene sulfonic acid (TNBS). Meanwhile, the effects of PSC-related factors, such as transforming growth factor ? 1 (TGF-? 1), collagen Ⅰ and MMP-2 on the pathogenesis of pancreatic fibrosis in rats were also evaluated. Methods Pancreatic fibrosis model in rats was induced by the injection of 2% TNBS in ethanolate-phosphate buffer solution into the pancreatic duct. The rats were sacrificed and the pancreata were removed at the 72nd hour, 3rd week, 4th week, 5th week, 6th week and 7th week after the operation respectively. Expressions of ?-smooth muscle actin (?-SMA), transforming growth factor ? 1 (TGF-? 1), collagen Ⅰ and MMP-2 were determined by either immunohistochemistry or RT-PCR, or Western blot respectively. The ultrastructure of pancreas was studied by electron microscope at different time points. Results The inflammation, swelling and necrosis were the major pathological changes of the pancreas at the early stage after the injection of 2% TNBS. Subsequently, the fibrotic manifestations such as proliferation of the fibrosis, atrophy of vesicles, deposition of collagen because prominent at the 3rd week after the operation, which peaked at 4th week. The expression of TGF-? 1 was increased significantly at the 3rd week after the operation and reached maximum at the 4th week. The expression of ?-SMA, which indicated the activation of PSC, could be detected at the 3rd week and also reached the peak value at the 4th week. After wards, it was decreased gradually. During the first 72 hours, the expression of MMP-2 mRNA was increased significantly and then was fluctuated but still higher than that in normal rats. The deposition of type Ⅰ collagen was increased in the areas of fibrotic tissues. Conclusions PSC might involve in the courses of the development and progression of TNBS induced pancreatic fibrosis in rats. This action was achieved via the activation of PSC by TGF-? 1, the production of those extracellular matrix metabolic associated enzymes such as the synthesis of collagen Ⅰ and the secretion of MMP-2.

BACKGROUND:Cervical spondylosis refers to cervical intervertebral spondylotic myelopathy and secondary degenerative changes, as wel as pathological changes in surrounding tissue structures. Establishing animal model of cervical instability and vertebral-basilar artery ischemia is the key in the studies addressing cervical spondylosis pathophysiology and treatment.
OBJECTIVE:To establish animal model of unstable cervical spine and vertebral-basilar artery ischemia, and explore new progress of animal model imitation study.
METHODS:A computer-based retrieval of PubMed database and CNKI database was performed for articles published from 1979 to 2012. The key words were“cervical instability, basal-vertebral artery ischemia, animal model”in English and Chinese. The articles about cervical instability, basal-vertebral artery ischemia, and animal model were screened, and those published recently or in authorized journals were preferred in the same field. Final y 43 articles were included in this study.
RESULTS AND CONCLUSION:An ideal animal model of cervical disease is needed. Animal model of cervical diseases is often used for the study of disease causes, onset mechanism and biochemistry. As the causes and mechanism of cervical diseases remain unclear, the existing modeling method cannot duplicate human cervical diseases, so further studies are needed to explore the establishment of models, positive rate and modeling time.

Objective To evaluate the value of 16-slice spiral computed tomography angiography(16SCTA) and reconstruction technique in the diagnosis of vascular diseases. Methods Using 0.75 mm collimation,1mm slice thickness and 0.5mm repitition,16SCTA was performed in 100 patients with suspected vascular diseases.Reconstruction methods were multiple planar reconstruction(MPR),maximum intensity projection(MIP) and volume rendering technique(VRT). Results 65 patients were proved by pathology or DSA in 100 vascular diseases,which included cerebral aneurysm(n=8),brain arteriovenous malformation(AVM,n=2),pulmonary AVM(n=6),pulmonary artery embolism with thrombus of the lower limb vein(n=9),pulmonary sequestration(n=4),coronary artery soft plaque and / or calcification(n=20),aortic aneurysm(n=12),renal artery stenosis(n=3),superior mesenteric artery thrombus(n=2),femoral artery stenosis(n=1),soft tissue hemangioma(n=33).Three reconstruction techniques showed the size,shape and extent of the lesion and displayed the lesion in any directions.VRT could display three-dimensionally the lesion.MPR and MIP could show the thrombus of lesion,MIP and VRT could display the calcification of lesion. Conclusion Application of every 16 SCTA reconstruction technique can display clearly lesion and replace DSA in diagnosis of vascular diseases,and provides another reliable diagnostic method for patients and has clinical importance for treatment.

Objective To evaluate the indication,operation pattern and therapeutic effect of ano-saving surgery for lower rectal carcinoma. Methods Retrospective analysis on the clinical feature of 320 patients with lower rectal carcinoma (postoperative time ≥5 years)treated by ano-saving surgery, the 5-year survival rate, local recurrence rate, and mortality were compared in the various operations. Results The success performed rate of ano-saving operation for lower rectal cancer was 58.5%(320/547).Among them, anastomotic leakage after surgery occurred in 4 cases (1.25%), and 26 cases had anastomostic narrowness (8.13%) within 1 year after surgery.The defecation function after surgery, in patients received colonic J pouch or transverse coloplasty pouch was much better than that in patients received coloanal or colorectal anastomosis. 5-year survival rates, and anastomostic recurrence rates were as follows:In ultra-low anastomosis were 63.24% and 10.27%. Park′s operation 66.67% and 5.13%, local resection 89.46% and 10.71%, respectively. 5-year recurrence rate in the pelvic soft tissue was 3.44%(11/320).Two cases died after operation. Conclusions Lower or ultra-low colon-rectum anastomosis becomes the main operative pattern in preserving anal sphincter in lower rectal cancer.Local resection of lower rectal tumor might be considered if the indecation is selected strictly. Colonic-J-pouch or transverse coloplasty pouch is good for improving the defecation function after ano-saving surgery for lower rectal cancer.

Objective To evaluate the effects of sitagliptin on blood glucose,blood pressure,blood lip and carotid artery intima media thickness (IMT) in metabolic syndrome patients with type 2 diabetes.Methods The clinical data were collected for 64 cases of inpatient and outpatient patients with metabolic syndrome with type 2 diabetes.Those patients included anti-diabetes native patients and patients only used the stable metformin dose.After signed off the informed consent form,those patients were randomized to the sitagliptin treatment group or original treatment group,and the metabolic index and carotid artery intima-media thickness were evaluated after 24 weeks treatment.Results The body mass index (BMI),waist circumference (WC),fasting plasma glucose (FPG),triglycerides (TG),high density lipoprotein cholesterol (HDL-C),systolic blood pressure (SBP),diastolic blood pressure (DBP),glycated hemoglobin a1c (HbA1c),and carotid artery IMT in two groups were comparable at baseline.After 12 weeks treatment,the FPG,TG,DBP,and HbA1c in the sitagliptin group were significantly better than original treatment group and the baseline,while there was no different between two groups in other index.After 24 weeks treatment,the FPG,TG,HDL-C,DBP,HbA1c,and carotid artery IMT in the sitagliptin group were significantly better than original treatment group and the baseline.Conclusions Sitagliptin presents the functions of lowering blood pressure,adjusting blood lipid,and protecting vascular endothelial in addition to lowering blood glucose.

ObjectiveTo investigate the incidence and risk factors of antibiotic-associated diarrhea (AAD) of pediatric patients with severe bacterial pneumonia.MethodsClinical data of 1086 pediatric patients with severe bacterial pneumonia from January 2010 through January 2014 were recruited. The incidence and risk factors of AAD were retrospectively analyzed. ResultsThe incidence of AAD in 1086 pediatric patients with severe bacterial pneumonia was 36.74%. The incidence of AAD in patients younger than 2 years old were higher than that in those older than 2 years, once or more times of mechanical venti-lation history were higher than that with no arrangements of this treatment, administering of combined antibiotics therapy were higher than that with single antibiotics, and the incidence of AAD due to amoxicillin/clavulanate, piperacillin/tazobactam, cefo- perazone/sulbactam in pediatric patients were 43.55%, 43.75%, and 45.03%, respectively. Three β-lactam/β-lactamase inhibitors above were risk factors of AAD through multivariate Logistic regression analysis.ConclusionThe high incidence of AAD in pediatric patients with severe bacterial pneumonia was associated with some risk factors, including younger than 2 years old, me-chanical ventilation, combined antibiotics therapy and administration of β-lactam/β-lactamase inhibitor (amoxicillin/clavulanate, piperacillin/tazobactam, cefoperazone/sulbactam).

Objective To examine mutations in the WT1 and PLCE1 gene in three Chinese families with autosomal recessive steroid-resistant nephrotic syndrome (SRNS) once mutations in NPHS2 had been excluded. Methods Peripheral blood samples were collected for genetic analysis from three probands of three Chinese families and their parents, and two probands' siblings, and 50 adult volunteers with normal urinalysis. Genomic DNA was isolated from peripheral blood leucocytes. Ten exons and exon-intron boundaries of WT1, and 31 exons and exon-intron boundaries of PLCE1 were amplified by polymerase chain reaction (PCR). Mutational analysis was performed by DNA sequencing directly and RFLP (restriction fragment length polymorphism) and/or PCR. Results No mutation in both WT1 and PLCE1 was identified in three probands from three Chinese families with autosomal recessive SRNS. However, three variants of WT1, 126C>T, ⅣS5-64A>G and 903A>G, and 13 variants of PLCE1, -134A>G, 810T>C, 960G>A, ⅣS11-28C>G, ⅣS15+26A>C, 4724G>C, ⅣS20+40C>T, ⅣS21+64G>A, ⅣS22-26T> A, 5320C>T, 5780A>G, ⅣS27+24A>G and ⅣS31 +48_49insT, were detected in three probands and some controls, indicating that all these variants were gene polymorphisms. WT1 polymorphism ⅣS5-64A>G, and PLCE1 polymorphism ⅣS22-26T>A were novel. Conclusion All the encoding exons and exon-intron boundaries of both WT1 and PLCE1 in three probands are examined, and no causative mutations in WT1 and PLCE1 axe found, suggesting that mutation in WT1 and PLCE1 genes is not a major cause of the Chinese families with autosomal recessive SRNS.

AIM:To approach the therapeutic effects of mycophenolate mofetil(MMF)in hormonal resistance nephrotic syndromes.METHODS:Patients were diagnosed as nephrotic syndrome and treated with prednisone at the dose of 1 mg?kg-1?d-1 for over 8 weeks,and 24 patients with unsatisfactory results or were palindromic were selected,and several patients in the 24 patients had been treated with cyclophosphamide or cyclosporine A.All patients were treated with MMF combined with low dose hormone.The initial dose of MMF was 1.0-1.5 g/d for 3 months,later the dose were reduced,and the maintenance dose of MMF was 0.5-1.0 g/d,the dose of prednisone was 5-20 mg/d,the follow-up visit period more than six months.The changes on urine protein,serum albumin,liver function,renal function were compared before and after treatment.RESULTS:Before and after treatment,urine protein decreased from(3.4?1.7)g/d to(0.9?0.2)g/d,serum albumin increased from(19.6?5.4)g/L to(36.1?7.7)g/L.serum creatinine level decreased from(105.7?6.4)?mol/L to(90.1?5.8)g/L.20 patients(83.3%)pathogenetic condition were relieved,15 patients(65.2%)were with complete remission.5 patients(20.8%)were partially recovered,and 4 patients(16.6%)had no response.The adverse effects were observed,including gastrointestinal events(n=8,33.3%),bacterial pneumonia(n=4,16.6%),herpes zoster(n=1,4.1%),hepatic function mild damage(n=3,12.5%).CONCLUSION:It is safe and effective to combine MMF with low dose hormone in treatment of hormonal resistance nephrotic syndrome,which could become a therapeutic option for refractory nephrotic syndrome(RNS).

BACKGROUND:There are a variety of treatments for femoral head necrosis,but their efficacy is not confirmed and unified.How to improve the differentiation ability of osteoblasts in the femoral head and improve the biomechanical support after the repair of the femoral head is an urgent problem to be solved.OBJECTIVE:To explore the clinical outcome of stem cells combined with vascularized iliac bone flap and tantalum rod implantation for the treatment of osteonecrosis of the femoral head (ONFH).METHODS:Totally 28 cases (36 hips) of non-traumatic ONFH admitted at the Zhongshan Hospital of Dalian University from January 2010 to January 2011 were enrolled.Bone marrow samples were extracted from each patient to isolate bone marrow stromal stem cells which were cultured in vitro for 2 weeks.Tantalum rod implantation with vascularized iliac bone graft was conducted to restore the femoral head shape,and then,prepared stem cell suspension were injected into the iliac bone flap and into the subchondral space of the femoral head.RESULTS AND CONCLUSION:All the 28 cases (36 hips) were followed up for 6-20 months (average 12 months),and their Harris hip scores and visual analogue scale scores at postoperative 6 and 12 months were significantly higher than the baseline (P ＜ 0.05).The Harris hip score at postoperative 12 months was significantly higher than that at postoperative 6 months (P ＜ 0.05),but there was no significant difference in the visual analogue scale scores at 6 and 12 months postoperatively (P ＞ 0.05).At the end of 12-month follow-up,clinical outcomes were excellent in 13 hips,good in 15 hips,fair in 4 hips,and poor in 4 hips,with an excellent and good rate of 90％.These findings indicate that autologous bone marrow stromal stem cell transplantation with vascularized iliac bone flap and tantalum rob implantation is an effective method with high clinical success rate for the treatment of ONFH.