Objective To explore the role of tissue inhibitor of metalloproteinase-1(TIMP-1) during renal senescence by using human TIMP-1 transgenic mice.Methods Renal histological changes of wild type mice and transgenic mice at the age of 3,12,24 months were observed by periodic acid-schiff(PAS)staining of paraffin sections.The numbers of F4/80 positive cells were detected by immunofluoreseence.The protein expressions of TIMP-1,TIMP-2,matrix metalloproteinase(MMP)-9,MMP-2,intercellular adhesion molecule-1(ICAM-1),transforming growth factor?1(TGF-?1),collagenⅢand collagenⅣwere detected by Western blot.The activities of gelatinases and TIMP-1 were examined by gelatin zymography and reverse zymography respectively.Results Focal renal fibrosis was found in two genotypes with aging.At the age of 24 months,compared with wild type,in kidneys of transgenic type,the expressions and activities of gelatinases were dowregulated (MMP-2:2.08?0.20 vs.3.39?0.43;MMP-9:4.02?0.82 vs.6.72?1.40,all P＜0.05);the expressions of collagenⅢ,collagenⅣ,ICAM-1,and TGF-?1 were upragulated(0.72+0.11 vs.0.57?0.09;0.84?0.13 vs.0.6?0.11,0.72?0.12 vs.0.53?0.07; 0.69?0.12 vs.0.45?0.09,all P＜0.05),and the numbers of F4/80 positive cells were increased (18.8?4.4 vs.12.7?3.6,P＜0.05)with the upregulated expression and activity of TIMP-1(1.10?0.18 vs.0.62?0.09;50.75?7.25 vs.20.64?3.50,P＜0.05).Conclusions TIMP-1 could promote age-related renal fibrosis through enhancing inflammation reaction by ICAM-1 upregulation.

A practical approach to the synthesis of the A, B and C-ring subunit of cyclopamine has been developed. This synthetic tactic highlights the utility of mandelate acetal-mediated resolution of the fused ring ketone (±)-4 and IBX-mediated oxidation cascades from 12 to 9. The availability of advanced intermediates from enantiomerically pure (+)-4 and 2 could provide efficient access to biologically active and structurally diverse C-nor-D-homo-steroidal alkaloids such as cyclopamine.
Chemistry Techniques, Synthetic ; methods ; Molecular Structure ; Organic Chemistry Phenomena ; Stereoisomerism ; Steroids ; chemistry ; Veratrum Alkaloids ; chemical synthesis ; chemistry

OBJECTIVE: To investigate the outcome of the kidney transplant recipients with different grades and stages of chronic hepatitis B virus after receiving renal transplantation for 3 years. METHODS: Thirty nine cases of kidney transplant recipients with hepatitis B virus and 20 cases of kidney transplant recipients (control group) between August 2000 and February 2002 were studied. Before the transplantation, the patients were divided into 4 groups: the mild hepatitis group (Group A, n = 8), the middle hepatitis group (Group B, n = 6), the severe hepatitis group (Group C, n =5) according to pathological diagnosis by percutaneous liver biopsy, and the control group (Group D). During the 3 year follow-up, the serum creatinine, alanine aminotransferase, g-Glutamyl transferase (GGT), total bilirubin, direct bilirubin, prothrombin time, cyclosporine trough concentration, urinary protein excretion, the HBV markers, HBV-DNA, albumin (A), globulin (G), the hepatic fibrosis markers and Child-Pugh score were studied at intervals. All patients received ultrasound examination every year. Two patients received repeated liver biopsy at the end of the follow-up in the hepatitis groups. RESULTS: The outcome of Group A and Group D was fine. In Group B, GGT level was significantly elevated (P < 0.05) sixth months after the operation, the Child-Pugh score of 2 patients were B, the liver pathohistological changes in another 2 patients were in severe stage in the endpoint. In Group C, GGT values had higher base-line (P <0.01) during the follow-up. Albumin were lower and globulin were higher than normal at the beginning of the 24th month. At the end of the follow-up, the Child-Pugh scores of all patients were B or C (B = 3, C = 2), 4 patients had end-stage cirrhosis, one died of hepatic cancer and the survival rate was 40% in Group C. CONCLUSION The outcome of the 3 groups is different. The pathohistological diagnosis by liver biopsy is important for patient selection receiving renal transplantation.
Adult ; Aged ; Female ; Follow-Up Studies ; Hepatitis B, Chronic ; complications ; surgery ; Humans ; Kidney Failure, Chronic ; complications ; surgery ; Kidney Transplantation ; Male ; Middle Aged ; Prospective Studies ; Treatment Outcome

Objective In order to study the localization of NMDA receptor protein on the neuron membrane, a new immunocytochemistry method to visualize and quantify 3D earmark the membrane protein in nanometer scale has been achieved by atomic force microscope (AFM) with labeled colloid gold particle. Methods First step, the distribution of the anti-NMDAR1 IgG-SPA-gold complexes molecule on the surface of mica had been scanned by AFM and the characteristic 3D conformation was analyzed by particle software. Then the 3D conformation was contrasted with the IgG-SPA-gold complexes molecules combined with the neuron membrane surface and that were compared with the results of light microscope, laser confocal microscope and scanning electron microscope. Results The 3D conformation of IgG-SPA-gold scanned through mica surface shows the average diameter of globe particles is 49 nm. The neuron membrane NMDA receptor protein combined with IgG-SPA-gold immunocomplexes are globe or cosh shapes . Its average diameter is 53 nm. The long axis section is a typical two apexes structure. Conclusion AFM could determine the situation and 3D conformation of NMDAR protein on neuron membrane surface in nanometer scale. NMDAR single molecule could combine with one or two colloidal gold particles. Colloidal gold particle scanned by AFM in situ is a new detecting method of immunocytochemistry.

Special designed group I intron ribozymes can specifically splice objective RNA, repair the mutant gene in RNA level. The specificity of ribozyme is determined by nucleotides specific internal guide sequence (IGS) introduced to the enzyme. In this study, fragment sequence containing Tetrahymena thermophilia intron I of 26S rRNA gene was cloned and cis-splicing activity of this ribozyme was confirmed by in vitro transcription. For evaluating the trans-splicing activity of this ribozyme, a truncated mutant Green Fluorescence Protein (GFP) vector, XYQ5/XYQ10- pEGFP-C2, was constructed. This vector deleted the 3' end 564bp fragment of EGFP coding sequence, led to the lost the activity of emitting green fluorescence. Trans-splicing ribozyme plasmids ptrans-rib-CMV2 for remedy of the truncated mutant EGFP was constructed by PCR and molecular cloning techniques. This vector utilizing cloned 26S rRNA intron 1 as core enzyme; selecting T-G site at 194bp of EGFP coding sequence as splicing receptor, designed an IGS which is inversely complement to the 188-193nt of EGFP mRNA; the 195-890bp fragment of EGFP coding sequence was ligated to the 3'-end of ribozyme core. The fragment containing these components was inserted to a eukayotic expression vector pRC-CMV2. Using linearized XYQ5/XYQ10- pEGFP-C2 and ptrans-rib-CMV2 as templates, truncated EGFP mRNA and the constructed ribozyme vector were transcribed and mixed to evaluate the trans-splicing activity. Analysis of in vitro transcription products mix by RT-PCR verified the existence of wild type EGFP mRNA molecule. Co-transfection of XYQ5/XYQ10- pEGFP-C2 with ptrans-rib-CMV2 to Hela cells proved this ribozyme restored green fluorescence within cell, but the efficiency was low.
Animals ; Base Sequence ; Green Fluorescent Proteins ; genetics ; HeLa Cells ; Humans ; Introns ; genetics ; Molecular Sequence Data ; Mutant Proteins ; genetics ; Mutation ; RNA, Catalytic ; genetics ; RNA, Messenger ; genetics ; Tetrahymena ; enzymology ; Trans-Splicing ; Transcription, Genetic

OBJECTIVETo provide scientific methods for quality criterion by studying the chemical components of essential oil from Baeckea frutescens.

METHODThe chemical components of essential oil from B. frutescens were identified by GC-MS-DS, TLC and capillary GC. The relative contents of main components were determined by area normalization.

RESULTMore than 50 peaks were separated, and 38 components were identified, which accounted for over 94% of the total GC peaks areas of the essential oil. The methods for quality evaluation of essential oil from B. frutescens by TLC and capillary GC were established.

CONCLUSIONThe chemical components of essential oil from B. frutescens collected from different habitats and collecting periods have common characteristics as well as differences. Some components, such as linalool, can be used as a standard and chromatography fingerprint to analyze the quality of essential oil from B. frutescens.

Bridged Bicyclo Compounds ; analysis ; China ; Cyclohexanols ; analysis ; Ecosystem ; Monoterpenes ; analysis ; Myrtaceae ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Seasons

OBJECTIVETo investigate the clinical application and indication of endoscopic dissection technique for submucosal tumors (SMTs) of the esophagogastric junction (EGJ) originating from the muscularis propria.

METHODSA total of 143 SMTs of the EGJ were treated by endoscopic resection in the Endoscopy Center of Zhongshan Hospital Affiliated to Fudan University between March 2007 and June 2011. The clinical and histopathologic feature, surgical approach, en bloc resection rate, complications, and postoperative follow up were evaluated.

RESULTSThere were 74 males and 69 females with a mean age of 49.1 years old. The en bloc resection rate was 94.4%(135/143). There were 126 patients who underwent endoscopic submucosal excavation in an en bloc fashion. Six patients underwent endoscopic full-thickness resection without laparoscopic assistance. Three patients underwent submucosal tunneling endoscopic resection. The other 8 SMTs were partially resected for histological evaluation and the residual tumors were further treated with nylon snare ligation. The mean lesion size was 17.6 mm. The mean procedure time was 45.1 minutes and the mean intraoperative bleeding was 50.0 ml. Perforations occurred in 6 patients and metal clips were used to close the defect. One patient with Mallory-Weiss syndrome was successfully treated with conservative treatment. Pathological examination showed that the lesions were leiomyoma (n=121), gastrointestinal stromal tumor (n=20), granulosa cell tumor (n=1), and intermuscular lipoma (n=1). No local recurrence and distant metastasis were noted during the follow-up (range, 3-48 months).

CONCLUSIONEndoscopic resection technique is safe and effective, and should be selected for each patient individually.

Adenocarcinoma ; surgery ; Adult ; Aged ; Endosonography ; Esophagogastric Junction ; pathology ; Female ; Follow-Up Studies ; Gastroscopy ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo prospectively evaluate the clinical value of different magnifying chromoendoscopy(MCE) methods in screening gastric precancerous lesions and early cancers.

METHODSBetween March 2010 and October 2011, among all the patients aged over 40 who received esophagogastroduodenoscopy at Zhongshan hospital, Fudan University, suspicious lesion was detected in 699 patients, who were randomly assigned to three groups: epinephrine dye(n=240), indigo carmine dye(n=246), and acetic acid-indigo carmine mixture dye(n=213). Diagnosis was made according to surface patterns and microvessels of the lesion. Pathological diagnosis was used as the gold standard. The concordance between endoscopic diagnosis and pathological diagnosis was evaluated through the agreement(Kappa) test. McNemar Paired chi-square test was used to compare the concordance of three MCE methods, regular white light, magnification alone, and NBI magnifier before and after MCE.

RESULTSPathological examination showed inflammatory lesions in 415 patients, intestinal metaplasia in 190, low grade intra-epithelial neoplasia in 17, and high grade intra-epithelial neoplasia or early cancer in 77. The percentage of patients with consistent endoscopic and pathological diagnosis was 77.1%(185/240) for epinephrine dye, 80.5%(198/246) for indigo carmine dye, and 81.2%(173/213) for acetic acid-indigo carmine mixture dye. Kappa values were 0.579, 0.502, and 0.667 respectively(all P<0.01). For the screening of high grade intra-epithelial neoplasia or early cancer, the diagnostic sensitivities were 84.0%, 83.3%, and 92.9%, respectively, and the specificities were 98.6%, 97.3%, and 98.4%. All the three chromoendoscopy methods improved the diagnostic accuracy for precancerous lesions compared with conventional gastroscopic observation with white light(all P<0.01). Indigo carmine and acid-indigo carmine mixture dye improved the diagnostic accuracy of magnification alone(both P<0.05). There was no significant difference in diagnostic accuracy between each MCE method and magnifying NBI observation(all P>0.05).

CONCLUSIONNBI magnification and all the three MCE methods may improve the diagnostic accuracy of early gastric cancer and precancerous lesions.

Acetic Acid ; Adult ; Aged ; Aged, 80 and over ; Coloring Agents ; Epinephrine ; Female ; Gastroscopy ; methods ; Humans ; Indigo Carmine ; Male ; Middle Aged ; Precancerous Conditions ; diagnosis ; pathology ; Prospective Studies ; Sensitivity and Specificity ; Staining and Labeling ; methods ; Stomach Neoplasms ; diagnosis ; pathology

Objective To evaluate the anti-angiogenic effect of eukaryotic vector containing human VEGF 165 and truncated pseudomonas exotoxin A (PE38) fusion gene on malignant glioma in nude mice, and explore a novel anti-angiogenic therapy for cancer. Methods The models were established through hypodermic injection of human U251 glioma cells into the nude mice and randomly divided into untreated group, PBS group, pIRES2-EGFP group and pIRES2-VEGF165-PE38-EGFP group on the 9th day. H&E staining was performed to observe the morphological changes of the glioma tissues; SP immunohistochemistry was employed to detect the expression of GFAP, CD31 andPE; quantitative pathologic image analysis system was used to investigate the microvessel density (MVD) in the tumor tissues. Results At day 16, the pIRES2-VEGF165PE38-EGFP group showed significantly lower tumor volume of mice and significantly decreased MVD than the other three groups (P<0.05). Positive expression of PE was shown in the pIRES2-VEGF165PE38-EGFP group, but negative in the other three groups. Conclusion The expression products of VEGFJ65-PE38 fusion gene can obviously inhibit the growth and angiogenesis of U2S1 cells in nude mouse flank tumor models, suggesting that it may be a novel therapeutic approach for anti-angiogenic therapy of cancer.

Objective To study the relationship between ischemic stroke (IS) and both gene polymorphisms of methylenetetrahydrofolate reductase (MTHFR) and cigarette smoking. Methods Four hundred and fifty-four patients with IS and 334 controls were recruited in our study; their gene polymorphisms of MTHFR were detected by PCR and denaturing high performance liquid chromatogram (DHPLC). The patients were further divided into different subgroups based on TOAST criteria and scores of neurological impairments and the distribution of MTHFR genotypes were analyzed accordingly. The relationships between IS and both cigarette smoking and these genotypes were measured by odds ratios (ORs) and 95％ confidence intervals (95％CIs). The ORs and 95％ Cls were calculated by unconditional logistic regression. Results TT genotype and T allele were associated with LAA type and CE type,moderate type and severe type of IS. OR of TT genotype and T allele in smoking patients with IS were 4.393 and 2.359, respectively; but the OR ofCC genotype in smoking patients with IS was 0.353. On the other hand, the OR of all genotypes and alleles in non-smoking patients with IS were not significantly different as compared with those in controls. Conclusion Cigarette smokers with T alleles are likely to suffer IS, but those cigarette smokers with C alleles are not; and there exist interactions between cigarette smoking and MTHFR gene in the pathogenesis of IS.