Objective To investigate expression of E-cadherin (E-cad) and ?-catenin (?-Cat) in pancreatic carcinoma and its relationship with the biological behavior. Methods Immunohistochemistry staining (PicTure TM two steps method) was used to detect E-cad and ?-Cat expression in 47 specimens of paraffin-embeded pancreatic carcinoma and 17 normal pancreas around tumors. Results E-cad and ?-Cat were expressed by normal ductal and acinar cells with typical membranous staining at the intercellular junctions. 17 normal specimens of pancreas strongly expressed E-cad and ?-Cat. The loss and reduction rate of E-cad expression in pancreatic carcinoma was 53.2%(25/47). The significant relationship was observed between low E-cad expression and differentiation,high hyperplastic grade and metastasis (P0.05). There was a positive relationship between the expression of E-cad and ?-Cat in the same specimen (r=0.88,P

Objective To investigate the methods of isolation and culture in vitro and detect the surface markers of human umbilical cord mesenchymal stem cells.Methods Human umbilical cord Wharton' s jelly was separated and cut up as small as possible,and then cultured with α-MEM.Human umbilical cord mesenchymal stem cells could be obtained by culturing the tissue block adhered the bottle wall.And the cells were passaged at a certain density.The surface markers of human umbilical cord mesenchymal stem cells were detected by FACS when the cells were in Generation Three.Results Human umbilical cord mesenchymal stem cells were obtained from Wharton' s jelly conveniently,with fibroblast shape and stable proliferation and passage.CD29,CD44,CD105 were strongly expressed on human umbilical cord mesenchymal stem cells.But CD45,CD34,HLA-DR,HLA-G,CD80,CDs6 were not expressed.Conclusion Human umbilical cord mesenchymal stem cells can be obtained effectively from the culture of the tissue block,which provides a rich source of cells for tissue engineering.

BACKGROUND:Studies have reported that taurine has a certain therapeutic effect on the disease of various systems, such as nervous system, cardiovascular system, immune system and digestive system. The liver is the main place, also the important target organ, of taurine metabolism. Therefore, the relationship between taurine and hepatopathy has become a hot topic in recent years. OBJECTIVE:To investigate the influence of taurine on superoxide dismutase and malondialdehyde expression in the liver tissue of rat models of liver fibrosis induced by carbon tetrachloride. METHODS:Thirty male C57B/L rats of SPF grade were randomly and evenly divided into blank control, model and taurine groups. Rats in the blank control group were intraperitonealy injected with 100% peanut oil of 1 mL/kg, twice a week, in total 10 weeks. Rats in the model group were intraperitonealy injected with peanut oil of 1 mL/kg containing 20% carbon tetrachloride, twice a week, in total 10 weeks. Rats in the taurine group were intraperitonealy injected with peanut oil of 1mL/kg containing 20% carbon tetrachloride, twice a week, in total 10 weeks, and were intragastricaly administered taurine of 500 mg/kg per day starting from the 3rd week til the 10th week. RESULTS AND CONCLUSION:Compared with the blank control group, the serum levels of hyaluronic acid, laminin, typeⅢ procolagen, typeⅣ colagen, alanine aminotransferase, aspartate aminotransferase were significantly increased (P < 0.05), the level of superoxide dismutase in the liver tissue was lowered (P < 0.05), the level of malondialdehyde in liver tissue was significantly increased (P < 0.05), and liver index was increased (P < 0.05) in the model group. Pathological examination showed that there were necrosis of liver cels, fat vacuoles, fibrous tissue hyperplasia and inflammatory cel infiltration in the rats of the model group. Compared with the model group, the serum levels of hyaluronic acid, laminin, typeⅢ procolagen, typeⅣ colagen, alanine aminotransferase, aspartate aminotransferase were significantly lowered (P < 0.05), the level of superoxide dismutase in the liver tissue was significantly increased (P < 0.05), the level of malondialdehyde in the liver tissue was significantly lowered (P < 0.05), and liver index was significantly decreased (P < 0.05) in the taurine group. Pathological examination showed that there were no inflammatory cel infiltration, fat vacuoles, and fibrous tissue deposition in the liver tissue. The results indicate that taurine can decrease the contents of superoxide dismutase and malondialdehyde, and relieve the degree of liver fibrosis induced by carbon tetrachloridevia exerting its antioxidative effects.

Objective To establishing an immortal cell line of familial papillary thyroid carcinoma (FPTC), and explore a new approach for studying familial non-medullary thyroid carcinoma (FNMTC). Methods The specimen from a patient with FPTC was selected, separated, and the primary cells were cultured using DMEM/F12 medium (with TSH, T3, EGF and hydrocortisone). To inducing cell immortalization, the exogenous genes SV40T/TERT were transfected into cells by two ways. RT-PCR was used to detect the expressions of thyroid peroxidase (TPO), thyroid globulin (TG), thyroid stimulating hormone receptor (TSHR) and sodium/iodide co-transporter (NIS). Immunofluorescence method was used to detect the expressions of TPO and GPC3. In order to detect the genomic mutations, the peripheral blood DNA of the patient was extracted. The cell genome was detected. Results The FPTC cells adhered to the plate and showed an irregular polygon shape. The cells can stably grow for six months, FPTC-S (with SV40T transfected) passaged to p26, FPTC cells passaged to p23 and FPTC-ST (with SV40T/TERT transfected) passaged to p19. Both FPTC-S and FPTC-ST can stably express TPO, TG and TSHR in mRNA level. MLH1 R217C mutation existed in the peripheral blood of the patient, and BRAF V600E mutation existed in the primary cultured cells. Either the primary or the immortal cells showed MLH1 R217C mutation. Conclusion This study preliminarily established an immortal cell line of familial papillary thyroid carcinoma with MLH 1 R217C and BRAF V600E mutations. This cell line provides a research model for studying these mutations in FPTC.

Objective To study the effects of tacrolimus(Tac) concentrations on the number of NK cells and receptor expression in peripheral blood of renal transplantation receptors.Methods A total of 60 first-time kidney transplantation recipients in our institute from Dec.2007 to July 2009 were followed up.Tac maintenance immunosuppressive therapy was given to all recipients.The recipients were divided into low-concentration Tac group (6.84 + 1.72μg/L,n =30) and highconcentration Tac group ( 11.88 + 2.59 μg/L,n =30) according to concentrations of Tac.Twenty healthy volunteers served as controls.Before and 6 months after operation,concentrations of Tac were analyzed by using micro particle immunoassay chemiluminescent method.NK cells and their receptors (CD85j,CD158d,CD94 and NKG2D) were detected by using flow cytometry.The concentrations of soluble HLA-G5 were detected by using ELISA.Results The number of NK cells in lowconcentration Tac group and high-concentration of Tac group preoperatively was significantly reduced as compared with control group (P ＜ 0.05 ). The percentage and number of NK cells in low concentration Tac group and high-concentration Tac group at 6th month after operation were significantly reduced as compared with control group (P＜0.05).The number of NK cells in lowconcentration Tac group was significantly greater than in high-concentration Tac group (P＜ 0.05).There was no significant differende in the expression of CD85j,CD158,CD94 and NKG2D before operation between two groups(P＞0.05).The expression of CD85j and CD158d in two groups was increased,but that of CD94 and NKG2D was decreased at 6th month post-transplantation as comapred with that preoperation.In low-concentration Tac group,the expression of CD85j and CD158d was increased as compared with that in high-concentration Tac group (P＜0.05 ).Spearman correlation analysis revealed that the CD85j and CD158d expression had a positive correlation with sHLA-G5(P＜0.01 ),but the NKG2D had a negative correlation with sHLA-G5(P＜0.01 ).Conclusion There was correlation between the concentrations of Tac and NK cells count and NK receptors. Low concentrations of Tac can safely and effectively protect kidney function.The number of NK cells andtheir inhibitor receptors are increased in the recipients with low concentration of Tac.

Because of the complexity of the cathepsin B-like (CBL) family, an information on the biological and biochemical characteristics of individual CBL genes is lacking. In this study, we investigated the degradative effects of the recombinant HC58 protein isolated from Haemonchus contortus parasites on protein substrates over a broad pH range in vitro. This protein, which hydrolyzed the synthetic peptide substrates Z-FR-AMC and Z-RR-AMC, had characteristics of the cysteine protease class of proteins. In the acidic pH range, the isolated protein actively degraded hemoglobin (Hb), the heavy chain of goat immunoglobulin G, and azocasein. By contrast, it degraded fibrinogen in the alkaline pH range. These activities were strongly inhibited in the presence of the cysteine protease inhibitor E-64. While the protein digested Hb, it did not induce the agglutination of erythrocytes from its natural host. These results suggest that the HC58 protein may play a role in the nutrition of this parasite.
Animals ; Caseins/metabolism ; Cathepsin B/antagonists&inhibitors/*genetics/isolation & purification/*metabolism ; Cysteine Proteinase Inhibitors/pharmacology ; DNA, Complementary/genetics ; Goat Diseases/*parasitology ; Goats ; Haemonchiasis/parasitology/*veterinary ; Haemonchus/*enzymology/genetics/isolation & purification ; Hemagglutination Tests/veterinary ; Hemoglobins/metabolism ; Hydrogen-Ion Concentration ; Immunoglobulin G/metabolism ; Leucine/analogs & derivatives/pharmacology ; RNA, Helminth/chemistry/genetics ; Recombinant Proteins/genetics/metabolism ; Reverse Transcriptase Polymerase Chain Reaction/veterinary

The complete coding sequence of Haemonchus (H.) contortus HC29 cDNA was generated by rapid amplification of cDNA ends in combination with PCR using primers targeting the 5'- and 3'-ends of the partial mRNA sequence. The cloned HC29 cDNA was shown to be 1,113 bp in size with an open reading frame of 507 bp, encoding a protein of 168 amino acid with a calculated molecular mass of 18.9 kDa. Amino acid sequence analysis revealed that the cloned HC29 cDNA contained the conserved catalytic triad and dimer interface of selenium-independent glutathione peroxidase (GPX). Alignment of the predicted amino acid sequences demonstrated that the protein shared 44.7~80.4% similarity with GPX homologues in the thioredoxin-like family. Phylogenetic analysis revealed close evolutionary proximity of the GPX sequence to the counterpart sequences. These results suggest that HC29 cDNA is a GPX, a member of the thioredoxin-like family. Alignment of the nucleic acid and amino acid sequences of HC29 with those of the reported selenium-independent GPX of H. contortus showed that HC29 contained different types of spliced leader sequences as well as dimer interface sites, although the active sites of both were identical. Enzymatic analysis of recombinant prokaryotic HC29 protein showed activity for the hydrolysis of H2O2. These findings indicate that HC29 is a selenium-independent GPX of H. contortus.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary/genetics/isolation & purification ; Glutathione Peroxidase/*genetics/*metabolism ; Goat Diseases/parasitology ; Goats ; Haemonchiasis/parasitology/prevention & control/*veterinary ; Haemonchus/*enzymology/*genetics ; Hydrogen Peroxide/metabolism ; Molecular Sequence Data ; Phylogeny ; RNA, Helminth/chemistry/genetics ; Random Amplified Polymorphic DNA Technique ; Rats ; Rats, Sprague-Dawley ; Sequence Alignment ; Sequence Analysis, DNA

Vol. 7, No. 3, pp. 249-255, 2006. The correspondence is changed by authors' request.

Objective To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) on the expression of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in mice with ischemia-reperfusion acute kidney injury (IR-AKI). Methods All mice were randomly divided into the sham operation group (control group), ischemia-reperfusion injury group (IRI group) and BMSC treatment group (BMSC group), with 6 mice in each group, respectively. The renal function and pathological changes of mice were detected. The cell apoptosis of renal tissues of mice was determined. The expression levels of serum IL-10 and TNF-α of mice were quantitatively measured. The mouse BMSC was randomly divided into the control and hypoxia-reoxygenation groups (IRI group), and the expression levels of IL-10 and TNF-α in cell supernatant were determined. Results The renal structure of mice was normal in the control group, severe damage was observed in the IRI group, and mild damage occurred in the BMSC group. Compared with the control group, the renal tissue injury scores were significantly higher in the IRI and BMSC groups (both P < 0.05). Compared with the IRI group, the renal tissue injury score was significantly lower in the BMSC group (P < 0.05). Compared with the control group, the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were remarkably up-regulated in the IRI group, and the level of BUN was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of Scr and BUN were significantly down-regulated in the BMSC group (both P < 0.05). In the IRI group, the quantity of apoptotic cells in the renal tissues was considerably higher than those in the BMSC and control groups, and the quantity of apoptotic cells in the BMSC group was significantly higher than that in the control group (all P < 0.05). Compared with the control group, the levels of serum IL-10 and TNF-α were significantly up-regulated in the IRI group, whereas the level of serum TNF-α was significantly down-regulated and the level of serum IL-10 was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of serum IL-10 and TNF-α were significantly down-regulated in the BMSC group (both P < 0.05). The levels of IL-10 and TNF-α in the cell supernatant did not significantly differ between the IRI and control groups (P=0.080、0.627). Conclusions BMSC infusion may reduce the incidence of renal IRI and inflammation, probably via the mechanism of down-regulating TNF-α expression rather than up-regulating IL-10 expression.

The 26S proteasome is a proteolytic complex responsible for the degradation of the vast majority of eukaryotic proteins. Regulated proteolysis by the proteasome is thought to influence cell cycle progression, transcriptional control, and other critical cellular processes. A novel Schistosoma japonicum gene (GenBank Accession No. AY813725) proteasome alpha2 subunit (SjPSMA2) was cloned. Sequence analysis revealed that the ORF of SjPSMA2 gene contains 708 nucleotides encoding 235 amino acids, and the molecular weight was estimated to be 25.84 kDa. Real-time PCR analysis showed that this gene expressed in 7 d, 13 d, 18 d, 23 d, 32 d and 42 d schistosoma. The mRNA level of SjPSMA2 was lower in 7 d and 23 d schistosomulum than that in other stages. The SjPSMA2 cDNA fragment was subcloned into an expression vector pET28a(+) and transformed into E. coli BL21 (DE3) cells. After induction with IPTCQ the 30 kDa fusion protein was produced as included bodies. Western-blotting revealed that the fusion protein could be recognized by the rabbit serum anti-Schistosoma japonicum adult worm antigen preparation, and the protein in native could be detected. After immunization of BALB/c mice with the fusion protein, the reduction rates of worm counts and liver egg counts were 12.33% and 35.23%. ELISA results revealed that the vaccinated group showed a significant increase in the level of IgG antibody. This study provided an important basis for investigating the regulation mechanism of the proteasome during the development of Schistosoma japonicum.
Animals ; Antibodies, Helminth ; blood ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genes, Helminth ; Helminth Proteins ; genetics ; metabolism ; Immunization ; Liver ; parasitology ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Parasite Egg Count ; Proteasome Endopeptidase Complex ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Schistosoma japonicum ; genetics ; metabolism ; Vaccines, Synthetic ; immunology