Objective: To study the chemical structure of polysaccharide CDP 4 isolated from Cistanche deserticola Y.C.Ma. Methods: The chemical properties of CDP 4 were determined by using chemical method and spectrocospic method. Results: CDP 4 was composed of glucosyl group, with the ratio 1,4 linkage glc p ∶1,6 linkage glc p =3∶1,and its mean molecular weight 1.4 ?10 4. By means of methlylation analysis, complete acid hydrolysis analysis, NMR spectrum, the linkages and sequence information of CDP 4 were obtained. Conclusion: CDP 4 is a new linear glucan.

Objective To investigate the clinico-pathological characters of acute and chronic Aristolochic acid nephropathy, and analysis the pathological mechanism of chronic Aristolochic acid nephropathy.Methods 26 cases of aristolochic acid nephropathy diagnosed in our department were examined. They were divided into acute and chronic group by their pathological characters. Immunohistochemical staining for the expression of collagen III, PAI-1, TIMP-1 and PCNA was done in renal biopsy specimens.Results There were 11 acute cases and 15 chronic cases. Compared with acute cases, there were more female, longer duration of the medicine intake[ (142.3?52.7 months of chronic cases and 4.5 ? 2.7 months of acute cases), higher degree of hypertension[(156.7?32.4) mm Hg of chronic cases and 127.3?24.2 mm Hg of acute cases], 24 hour urinary protein,anemia, glomerulosclerosis, tubular atrophy, interstitial fibrosis and artery lesions in chronic patients(Pall

Objective To investigate the effects of shRNA produced by PCR on the suppression of the expression of exo- or endogenous genes. Methods The specific primers were designed, with which the shGFP-RNA and shFN-RNA were produced by PCR. The shGFP-RNA was transfected into 293T cell lines together with pEGFP plasmid, then the cells were detected by laser confocal microscopy 48h later. The shFN-RNA was transfected into rat mesenteric cell lines, then the cells were collected 48h later and the total RNA was extracted, which was reversely transcripted to cDNA. Then the expression level of FN mRNA was examined with real-time PCR, and the expression level of FN protein was examined with Western blot analysis. Results The results of laser confocal microscopy indicated that the EGFP could be successfully suppressed by shGFP-RNA produced by PCR; the results of real-time PCR and Western blot analysis revealed that FN expression level of the cells transfected with shFN-RNA was down regulated, and the level was much lower than those tansfected with independent shRNA (P

Objective To prepare the anti-hUART1 polyclonal antibody and investigate the subcellular localization of hURAT1 protein. Methods The full-length hURAT1 gene was obtained by RT-PCR and inserted into the fusion expression vector pEGFP-N3,the predicated antigen epitope was cloned into GST fusion protein expression plasmid pGEX-5X-1, transformed into E. coli BL21 cells for expressing recombinant GST-hURAT1 protein induced by IPTG. The purified hURAT1-GST fusion protein was employed to immunize rabbit for preparing the polyclonal antibody. The expression of hURAT1 was analyzed by western-blot and immunohistochemistry in human kidney. pEGFP-hURAT1 was transfected into the LLC-PK1 cell in order to observe the subcellular localization of the gene using confocal microscopy. Results Specific anti-hURAT1 rabbit polyclonal antibody was obtained, and both Western-blot and immunohistochemistry showed that hURAT1 was expressed in the human kidney brush border,localized in the apical membrane of the LLC-PK1 cell. Conclusions hURAT1 protein was a membrane protein located in renal proximal tubule, which could be detected in the apical membrane. The anti-hURAT1 polyclonal antibody could be used for studying the physiological function of hURAT1 and its pathology.

Objective To study the change of human Na+/dicarboxylate cotransporters (NaDC)in IgA nephropathy,and to discuss its role in IgA nephropathy. Methods Thirty-four cases of IgA nephropathy diagnosed by renal biopsy were divided into five grades according to the degree of pathological change of IgA nephropathy,and their tubulointerstitial lesions were semi-quantitatively analyzed. Changes of the expression of NaDC1 and NaDC3 in kidney were observed by using immunohistochemistry,and analyzed with clinical data by correlation. Results Tubulointerstitial lesion exacerbated with the progress of pathological change in IgA nephropathy. In the cases of Ⅳ and Ⅴ grade,24 h urinary protein,serum creatinine,urinary NAG enzyme were increased,and urinary osmotic pressure was decreased,as compared with the cases of Ⅰ-Ⅲ grade( P

Current reform of higher medical education focuses on curriculum integration and corresponding reconstruction of teaching system. Chongqing Medical University has carried out the reform of medical personnel training mode from basic to clinical, which has achieved certain results. Three aspects including teaching content, teaching methods and appraisal system of the urogenital system curriculum integration are introduced in the paper. In the teaching content, the parts related to urogenital system in basic subjects and clinical subjects are extracted and integrated. In the process of teaching implementation, a teaching team is set up across departments and multidisciplinary joint teaching is carried out. In order to arouse the enthusiasm of students, inspire and cultivate students' scientific research thinking by improving teaching methods, several teaching methods such as case introduction teaching method, question discussion and debate meeting are used. Scientific evaluation system is used to observe and record the whole process of students' learning. Meanwhile, some existing problems and solutions of curriculum integration are discussed in the paper to provide references for peers in relevant colleges and universities.

Objective To compare the screening effects between Wells and revised Geneva scores on suspected acute pulmonary thromboembolism (APTE),and to explore a optimum screening method for APTE in the emergency department of China.Methods The study was carried out by using random,crossed,prospective methods to compare the screening effects between Wells and revised Geneva scores for 167 suspected APTE patients in the emergency department of the First Affiliated Hospital of Xiamen University.Results The areas under the receiver operating characteristic curve of Wells and revised Geneva scores for screening APTE in the emergency department were (0.917 ± 0.022 ) and (0.927 ± 0.020),respectively ( P ＜ 0.05 ).The diagnostic concordance between the two score systems for predicting APTE was poor (Kappa value =0.276 ). In addition, the difference between their hierarchical discrimination for the possibility of APTE was statistically significant ( P ＜ 0.05 ).Compared with revised Geneva score,fewer patients were diagnosed with low clinical probability of APTE and more patients were diagnosed with intermediate or high clinical probability of APTE through Wells score.The patients with low chnical probability of APTE were excluded from pulmonary embolism in Wells or revised Geneva score.At intermediate clinical probability,the accuracy rate of Wells score for predicting APTE (9.64％) was lower than that (32.84％ ) of revised Geneva ( P ＜ 0.05 ).At high clinical probability,there was no significant difference between their accuracy rate [ (67.24％ vs.86.21％),P＞0.05]. Conclusions Revised Geneva score is more suitable than Wells score in screening suspected APTE patients in the emergency department in our country.

Objective To explore the mechanism of protecting cells from ischemic reperfusion injury by constructing specific small interference RNA (siRNA) to inhibit Na+-H+exchanger-1 (NHE-1) expression in human renal tubular epithelial cell (HKC). Methods The siRNA was designed and synthesized based on human NHE-1 complete sequence,and was transfected into HKC.The irrespective siRNA transfected group was used as control.The cells were treated with 10 μmol/L antimyein A to induce ischemia and anoxyaemia environment.NHE-1expression was examined by RT-PCR and Western blot.The intraeellular pH (pHi),Ca2+ or Na+ concentrations were detected by BCECF/AM,Fluo-3/AM and SBFI-AM,respectively,combining with laser eonfocal assay system.Nucleic morphology was determined by Hoechst 33342.Cellular apoptosis was examined by Annexin V/PI staining and flow eytometry.Fluorescent probe JC-1 was used to detect the change of mitechondrial transmembrane potential. Results The specific siRNA could efficiently inhibit NHE-1 expression in HKC.Compared with the irrespective siRNA transfected group,the mRNA and protein expression of NHE-1 was significantly down-regulated in NHE-1 siRNA transfeeted group (all P＜0.05).After treatment with antimyein A,the mRNA and protein expression of NHE-1 was significantly up-regulated in both groups,however,it was less than that in irrespective siBNA transfected group.At the same time,the ratio of apoptosis decreased (8.9% +2.9% vs 18.8%±3.2% , 17.4%±3.6% ,P＜0.05) and mitochondrial transmembrane potential rose significantly in NHE-1 siRNA transfected group as compared to irrespective siRNA transfected group and antimycin A group.The intracellular Na+,H+ and Ca2+concentrations increased in NHE-1 siRNA transfected group treated with antimyein A,but their levels were lower than those in irrespective siRNA transfected group with the same treatment(P＜0.05). Conclusions The synthesized siBNA can inhibit the expression of NHE-1 and can protect HKC from isehemia reperfasion injury induced by antimyein A.The mechanism might be via suppressing the expression of NHE-1 to delay intracelluar Na+ accumulation,attenuate intracellular Ca2+ overloading,and inhibit the decrease of mitechondrion transmembrane potential and reduce cellular apoptosis.

Objective To explore the clinicopathological features of IgA nephrolpathy associated with malignant hypertension (IgAN-MHT) and to analyze their correlation with renal vascular lesions. Methods Twenty-nine patients of IgAN-MHT were screened from 2000 biopsy-proven eases with primary IgA nephropathy (IgAN) in our department from April 1997 to May 2007. Data of clinicopathology and follow-up of these 29 patients were collected. Semi- quantitative analysis was performed to evaluate the pathological changes. Inner lumen, outer lumen, intimal thickness, tunica media-to-internal lumen ratio of 436 arterioles, 124 interlobular arteries and 5 arcuate arteries were measured. The primary endpeint was the composite of a doubling of serum creatinine level and ESRD. Correlations of renal vascular lesions with clinical manifestation, pathological change and prognosis were examined by Spearman and Cox methods. Results 1.5% of all the IgAN patients presented malignant hypertension. The common clinical features were renal failure (100%), hyperurieacidemia (62.7%) and hypertriglyceridemia (51.7%). The average amount of urine protein excretion was 2.8 g/d. The common pathological changes were moderate mesangial proliferation, severe global sclerosis, severe interstitial inflammation and severe interstitial- tubular fibrosis. The small arteries (arcuate arteries and interlobular arteries) and arterioles (afferent arterioles) were both involved in IgAN-MHT. The characteristic lesions of intrarenal arteries included vascular occlusion, media thickening, proliferative endarteritis (onionskin lesion, musculomucoid intimal hyperplasia), hyaline arteriosclerosis, but mainly vascular occlusion (86.2%). The arteriole lesion was negatively correlated with age and total protein level; vascular occlusion was positively correlated with uric acid level. The average foUow-up period was 21.1 months. Forteen patients reached the endpoint. The arteriole lesion was the main independent risk factor for the progression of IgAN-MHT (RR=10.21, 95%CI=1.16～89.67). Conclusions The main clinical feature of IgAN-MHT is renal failure. The main histological feature of intrarenal vascular lesions is occludes arterioles. Arteriole lesion is the main independent risk factor for the progression of IgAN-MHT.

Objective To determine the urinary polypeptide patterns of glomerulonephritis by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) technology. Methods Urinary samples of 29 healthy volunteers and 34 patients with glomerulonephritis, including 10 cases of IgA nephropathy (IgAN), 10 cases of membranous nephropathy (MN), 9 cases of minimal change disease (MCD) and 5 cases of lupus nephritis (LN), were collected and separated by magnetic bead,and were screened for polypeptide patterns with a novel high throughput method, MALDI-TOF MS. Results Under the relative molecular weights 10 000 Da, 85 protein peaks were detected in healthy controls group and 109 protein peaks were detected in glomerulonephritis group. Six peaks of 3371.5 Da, 4026.35 Da, 4085.32 Da, 4116.96 Da, 4126.32 Da and 9527.31 Da were up-regulated,while 8 peaks of 861.28 Da, 1205.41 Da, 1642.52 Da, 1913.15 Da, 1976.52 Da, 2087.74 Da, 2193.47 Da and 3015.57 Da were down-regulated by more than 2 folds (P<0.01) in glomerulonephritis group as compared to healthy controls. Urinary polypeptide patterns in different diseases differed significantly from each other, indicating specific disease pattern of polypeptide excretion. Conclusions MALDI-TOF MS is a fast, convenient and high throughput analyzing method capable of screening some relative specific, potential biomarkers from the urine of glomerulonephritis patients thus it possesses better clinical value.