Objective To observe the effect of astragaloside IV (A) and SB203580 antagonize cadmium (Cd) toxicity on expression of associated protein and blood-testis barrier(BTB) in rats and to study the protective mechanism of A on it.Methods Totally 21 SD male rats were randomly divided into 7 groups, 3 rats per group:Cd [ intraperitoneally injected with 0.1%CdCl2,1mg/(kg?d)],Cd+A [at the above dose of CdCl2,at the same time with A,10mg/(kg?d)], and Cd +SB203580 [at the above dose of CdCl2,at the same time with SB203580,100μg/(kg?d)], each of the above groups was further divided into continuous five and ten days treatment groups .The control group was intraperitoneally injected with equal dosage of normal saline .The testes were studied by light , electron microscopy , immunohistochemistry and Western blotting .Results In the control group ,irregular and lightly stained nuclei of Sertoli cell ( Sc) in seminiferous tubules were observed by HE staining .A continuous electron density line of tight junction ( TJ) and normal ultrastructure of BTB were observed .After Cd treatment ,the vesicular formation in the Sc was observed .The ultrastructural damage of Sc and TJ was observed .Compared with the corresponding time point of Cd group ,these were weakened in morphology of testis and ultrastructure of TJ after Cd +A or Cd +SB203580 treatment .The positive products of zonula occludens-1 ( ZO-1 ) and claudin-11 were localized mainly in the base of the seminiferous tubule .After Cd treatment , the average absorbance (AA) of ZO-1 and Claudin-11 was decreased significantly compared with that of the control group (P<0.05).After Cd +A or Cd +SB203580 treatment,AA of ZO-1 and Claudin-11 were increased significantly compared with that of the Cd group(P<0.05),though lower than that of the control group .The result of Western blotting showed that phosphorylation-p38MAPK in Cd group was increased significantly compared with that of the control group (P<0.05).After Cd +A or Cd+SB203580 treatment, it was decreased significantly compared with that of the Cd group (P<0.05).Conclusion Cd decreases ZO-1 and Claudin-11 expression and damages ultrastructure of TJ in BTB , asⅣhas protective effect on it , and is related to inhibiting activation of p 38 MAPK pathway .

Objective To investigate the toxic effect of cadmium (Cd) on the ultrastructure ,expression of related protein and the signal molecule phosphrylated P38 mitogen-actived protein kinase(P-P38MAPK) of primary cultured rat sertoli cell(Sc) ,and the protective effect of astragaloside (A ) on it .Methods The primary cultured rat Sc were divided into the control group ,Cd (50 mol/L)group and Cd(50 mol/L) plus A(10 mg/L) group ,they were used for the electron microscope observation and the im-munohistochemistry detection of vimentin ,E-cadherin ,-catenin and P-P38MAPK .Results The Sc ultrastructural changes included that the swelled mitochondria ,abundant lipid droplets and dilated endoplasmic reticulum were found in the Cd group .Further ,apop-tosis occurred in some Sc .However these ultrastructure changes above mentioned were slighter in the Cd plus A group ;the immu-nohistochemistry showed that the positive products of vimentin ,E-cadherin and-catenin were obviously decreased in the Cd group (P<0 .05) ,and those in the Cd plus A group were higher compared with the Cd group(P<0 .05);the expression of P-P38MAPK in cytoplasm was increased in the Cd group ,and showed the trend to move from cytoplasm to nucleus ,meanwhile ,the positive prod-ucts expression in the Cd plus A group was lower than that in the Cd group (P<0 .05) .Conclusion Cadmium can cause the injury of the Sc ultrastructure ,damage of cytoskeletal protein and fibronectin ,and increase of P-P38MAPK level ;astragaloside can antago-nize the toxicity of cadmium on Sc ,the protective effect maybe related with the decrease of P-P38MAPK in Sc .

Bacteria isolated from rotten hides,one identified as Pseudomonas aeraginosa later, were shown to decompose fresh hides completely in 48 hours at room temperature. Strongly collagenolytic activity was detected when native collagen from calfskin was used as substrate. The optimal reaction pH value and temperature of the collagenace produced by P.aeraginosa are 7.5 and 32℃ respectively. EDTA and EGTA inhabit the collagenolytic activity severely while PMSF has little effect on it. Not only media components but also fermentation conditions have differents effect on the production of this collagenolyic enzyme.

Immunohistochemical and electron microscopic techniques and image analysis were employed to investigate the regulation of cardiac AT1A and AT2 subtype receptors in rats during cardiac remodeling following myocardial infarction (MI). Positive immunostaining for AT1A and AT2 receptors was observed in myocytes and vessels with AT1A being more than AT2. Three days after MI, disappearance of myocardial cross striation and fibroblast hyperplasia were found with electron microscopy. AT1A receptor protein expression in myocardial noninfarcted portion notably increased compared with that in sham-operated control rats (P<0.001). No apparent changes were observed in AT2 receptor (P>0.05). Two weeks after MI, myocyte cross striation and collagen deposition were found. Meanwhile, AT1A receptor staining decreased compared with that of three days after MI (P<0.01), but there was still more than that of the control (P<0.05). AT2 receptor was significantly increased compared with that of the sham-operated control rats (P<0.001). These results suggust that both AT1A and AT2 receptor protein expression was upregulated in noninfarcted myocardium after MI, and the regulation of AT1A and AT2 receptors after MI may be involved in post-infarction cardiac remodeling.

BACKGROUND:The production and release of a large amount of inflammatory factors caused by immune system inflammatory response mainly contributes to secondary spinal cord injury. OBJECTIVE:To investigate the effects of umbilical cord Wharton’s jely mesenchymal stem cel transplantation on repair of injured neurological function and expression of inflammatory factors monocyte chemoattractant protein 1 and interleukin 10 in rats with acute spinal cord injury. METHODS: Eighty-one healthy adult male Sprague-Dawley rats were randomly and equaly divided into sham operation, model and cel transplantation groups, with 27 rats per group. Rats in the latter two groups were subjected to hemisection of the spinal cord to establish acute spinal cord injury models. Rat models in the cel transplantation group received umbilical cord Wharton’s jely mesenchymal stem cel injection (1×106)via the tail vein. Rat neurological function was evaluated using the BBB score at different time points after spinal cord injury. The expression of monocyte chemoattractant protein 1 and interleukin 10 in injured spinal cord tissue was detected using ELISA assay at different time points after spinal cord injury. Migration and neuronal differentiation of umbilical cord Wharton’s jely mesenchymal stem cels in the injured spinal cord tissue were determined using immunohistochemical staining method. RESULTS AND CONCLUSION:Compared with the sham operation and model groups, rat neurological function was significantly recovered in the cel transplantation group (P < 0.05). Compared to the model group, monocyte chemoattractant protein 1 level in the serum and monocyte chemoattractant protein 1 mRNA and protein expression in the injured spinal cord tissue were significantly lower (P < 0.05), but interleukin 10 mRNA and protein expression in the injured spinal cord tissue was significantly higher (P < 0.05), in the cel transplantation group. In the cel transplantation group, umbilical cord Wharton’s jely mesenchymal stem cels could migrate to the injured region and express glial fibrilary acidic protein. These findings suggest that umbilical cord Wharton’s jely mesenchymal stem cels promote rat neurological function recovery by regulating the inflammatory response in the injured spinal cord tissue, which is likely to be one of mechanisms by which transplantation of umbilical cord Wharton’s jely mesenchymal stem cels treats spinal cord injury.

Based on site-specific transposition of an expression cassette into a baculovirus shuttle vector (Bacmid) which propagated in Escherichia coli, the Bac-to-Bac System provides a rapid and efficient method to generate recombinant baculoviruses and is widely used for high level expression of heterologous proteins. And the efficiency of recombinant baculovirus infecting cells plays an important role on the protein expression. In this study, we introduced an EGFP expression cassette driven by polyhedrin promoter into the p74 locus of Bacmid by homologous recombination. The target Bacmid-egfp was then transformed into E. coli DH10B containing the transposition helper plasmid to gain a new transposition receipt strain E. coli DH10Bac-egfp. Because of the intact attTn7 sites and lacZ', target gene cloned in a pFastBac vector can be transposed into the Bacmid-egfp shutter vector to construct recombinant baculovirus, which would allow the tracing of the target protein expression and the recombinant Bacmid transfection or recombinant baculoviral infection under fluorescence microscopes. Recombinant virus Bac-egfp-DsRed was constructed by transposing DsRed into the Bacmid-egfp in E. coliDHl0Bac-egfp, and the Sf9 cells infected with the recombinant virus expressed DsRed and EGFP efficiently. Another protein IL-6 fused with 6 x his tag was expressed and purified sucessfully from Sf9 cells infected with recombinant virus Bac-egfp-6 x his-IL6 constructed by the improved Bac-to-Bac system.
Animals ; Baculoviridae ; genetics ; Green Fluorescent Proteins ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Spodoptera

OBJECTIVETo investigate the effect of the basic fibroblast growth factor (b-FGF) to regenerate an autologous tissue-engineered cartilage in vitro.

METHODSThe Cells were harvested from the elastic auricular cartilage of swine,and were plated at the concentration of 1 x 10(4) cells/cm2 , studied in vitro at two different media enviroments: Group I contained Ham's F-12 with supplements and b-FGF, Group II contained Ham's F-12 only with supplements. The passage 2 cells (after 12.75 +/- 1.26 days) were harvested and mixed with 30% pluronic F-127/Ham's F-12 at the concentration of 50 x 10(6) cells/ml. It was injected subcutaneously at 0.5 ml per implant. The implants were harvested 8 weeks after the vivo culture and examined with the histological stains.

RESULTSThe chondrocytes displayed morphologically similar to the fibroblasts in the media containing basic-FGF. The number of cell doublings (after 12.75 +/- 1.26 days) in vitro culture was as the following: Group I, 70; Group II, 5.4. Eight 8 weeks after the vivo autologous implantation, the average weight (g) and volume (cm3) in each group was as the following: Group I, 0.371 g/0.370 cm3 Group II, 0.179 g/0.173 cm3 (P < 0.01). With the b-FGF in vitro culture, the cells were expanded by 70 times after 2 weeks. Histologically, all of the engineered cartilage in the two groups were similar to the native elastic cartilage.

CONCLUSIONThese results indicate that the basic-FGF could be used positively to enhance the quality and quantity of the seeding cells for the generation of the well-engineered cartilage.

Animals ; Cartilage ; cytology ; drug effects ; physiology ; Cell Division ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; Female ; Fibroblast Growth Factors ; pharmacology ; physiology ; Male ; Regeneration ; drug effects ; Swine ; Tissue Engineering ; methods ; Transplantation, Autologous

OBJECTIVETo study the physiopathologic basis of Weikangfu Granule (WKFG) in treating precancerosis of gastric mucosa in patients of chronic gastritis with Pi-deficiency syndrome (CG-PDS).

METHODSOne hundred and fifteen patients of CG-PDS who suffered from intestinal metaplasia (IM) and atypical hyperplasia (ATHP) of gastric mucosa, were divided into two groups. The treated group (n = 61) was treated by WKFG with its ingredients modified according to the syndrome type of patients. The control group (n = 54) was treated with Weishu granule. The histopathological and subcellular ultrastructural changes were detected by optical microscope, screening electronic microscope, transmission electronic microscope and histochemical staining; the nuclear and mitochondrial ultrastructure of gastric mucosa were analyzed with energy dispersion X-ray analyser and image analysis system. And the changes of cAMP, lipid peroxide (LPO), superoxide dismutase (SOD) before and after treatment in the treated group were measured and compared with those of the health control group consisting of 15 volunteers.

RESULTSThe symptomatic and pathological therapeutic effect in the treated group were significantly superior to those in the control group (P < 0.05). The contents of Zn, Cu, cAMP, SOD and (3)H-TdR LCT in gastric mucosa of the treated group before treatment were all lower than those of the healthy control group, yet all these indexes markedly increased after treatment, while serum LPO level, which increased before treatment was lowered after treatment. All the changes showed statistical significance (P < 0.05 or P < 0.01).

CONCLUSIONWKFG can reverse IM and ATHP in patients of CG-PDS, and the effect may be realized by way of increasing the level of Zn, Cu, cAMP and SOD in gastric mucosa, promoting cell differentiation, enhancing cellular immunity and reducing oxygen free radicals and lipid peroxidation.

Adult ; Aged ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Chronic Disease ; Copper ; analysis ; Cyclic AMP ; analysis ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; chemistry ; pathology ; ultrastructure ; Gastritis, Atrophic ; pathology ; Humans ; Lipid Peroxides ; analysis ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Precancerous Conditions ; pathology ; Stomach Neoplasms ; pathology ; Superoxide Dismutase ; analysis ; Syndrome ; Yang Deficiency ; complications ; Zinc ; analysis

OBJECTIVETo study the expressions of matrix metalloproteinase 2 (MMP2) and E-cadherin (E-CD) in salivary mucoepidermoid carcinoma, and their relationship with clinical stages, pathological grading, lymph node metastasis and prognosis.

METHODSSurgical specimens of salivary mucoepidermiod carcinoma and normal salivary gland tissue were collected. MMP-2 and E-CD were stained immunohistochemically with streptavidin peroxidase method.

RESULTSThe expression of MMP-2 was increased and the expression of E-CD was reduced or negative in salivary mucoepidemoid carcinoma compared with those of the normal salivary gland. Expression of MMP-2 and E-CD was closely correlated with lymph node metastasis of the mucoepidermoid carcinoma. MMP-2 was positively correlated with the prognosis of mucoepidermoid carcinoma, and E-CD was negatively correlated to the prognosis of mucoepidermoid carcinoma.

CONCLUSIONThe expression of MMP-2 and E-CD is closely correlated with the metastasis and prognosis of salivary mucoepidermoid carcinoma.

Cadherins ; biosynthesis ; genetics ; Carcinoma, Mucoepidermoid ; metabolism ; pathology ; Humans ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Prognosis ; Salivary Gland Neoplasms ; metabolism ; pathology

BACKGROUNDMagnetic resonance imaging (MRI) is prone to be deformed by artifacts caused by the presence of metallic materials. The aim of this study was to evaluate the artifacts from galvano-ceramic and metal-ceramic crowns in MRI, in order to analyze their influences on diagnostic interpretation of MRI.

METHODSGalvano-ceramic and metal-ceramic crowns (Bio98, Wiron99, SP-78, BioKC97) were fabricated with the same model. All materials were imaged by means of 1.5T MRI apparatus with three different sequences, T1-weighted spin-echo (T1-weighted SE), T2-weighted spin-echo (T2-weighted SE) and Gradient echo (GE). Mean and standard deviation of distilled water signal intensity (SI) around the sample in the region of interest (500 mm2) enclosing the whole artifacts were determined, and compared for evaluation of the homogeneity of signal intensity. Images around the sample were acquired and evaluated.

RESULTSThere were statistically significant differences in the values of signal intensity between acrylic resin control and BioKC97, Wiron99 in the three sequences (P<0.001). The mean values of signal intensity for Bio98, SP-78 were significantly different from that of acrylic resin control (RE) in GE sequence (P<0.001). No difference was showed between acrylic resin control and galvano-ceramic crown (P>0.05). Images showed that the greatest artifact was a 25 mm ring with distortion in Wiron99 in GE sequence.

CONCLUSIONSThis in vitro study suggested that galvano-ceramic crown had no influence on the MRI, while metal-ceramic crowns caused moderate artifacts in the MRI. Therefore, galvano-ceramic restoration is a valuable alternative in dentistry.

Artifacts ; Ceramics ; Crowns ; Dental Alloys ; Humans ; Magnetic Resonance Imaging ; methods ; Metals