Aim To study the effect of gender differences in C57BL / 6J mice on antigen induced Sjogren's syndrome(SS)model. Methods The submandibular gland protein of C57BL/6J female and male mice was extracted and mixed with the same amount of Freund's complete adjuvant(FCA)for the first three times, the antigen concentration was adjusted to 2.5 g·L-1, mixed with Freund's incomplete adjuvant(FIA)for the fourth time, and the same-sex mouse antigen was injected into the back of mice for a total of four times to induce the mouse SS model. The mouse SS model was induced by multi-point intradermal injection of antigen on the back of mice for four times,the body weight of female and male mice was measured every week, the general condition was observed, the saliva volume of mice was measured at the sixth week of modeling. After the mice were sacrificed, the pathological changes of submandibular gland and the changes of T and B lymphocyte subsets in spleen were detected, and the differences in SS model preparation between female and male mice were compared. Results The SS model of male and female mice was successfully established, and there was no significant difference in general condition, saliva volume, submandibular gland pathology, plasma cells and memory B cells between male and female SS mice. The success rate of SS model was 75% in female mice and 60% in male mice. Compared with normal mice of the same sex, the weight loss of female SS mice was earlier and more obvious than that of male SS mice; the submandibular gland index of male mice was significantly higher than that of female mice. Compared with normal mice of the same sex, the proportion of Th17 and Treg cells in spleen of female SS mice was more statistically significant than that of male SS mice. Conclusions The success rate of SS modeling in female mice is higher than that in male mice. Compared with male SS mice, female SS mice have more significant SS like manifestations and pathological manifestations, which can provide a reference basis for the selection of gender when establishing SS model.

Aim To observe the therapeutie effeet of (AA) rats and its effect on the aetivity of tryptophan allopurinol ( ALLO) on adjuvant induced arthritis 2,3 dioxygenase (TDO).Methods SD rats were randomly divided into normal group, AA model group, ALLO group (10,20 , 40 mg • kg 1 ) and methotrexate group (0.5 mg • kg 1 ).The AA rats were established by intracutaneous injection of complete Freund's adju¬vant into the right toes of rats.The body weight,joint swelling number, joint pathology, spleen index and fi¬broblast like synoviocytes ( FLS) proliferation of the rats were observed to explore the therapeutic effect of ALLO.Flow cytometry detected the number of CD68 ∗ macrophages and the ratio of Thl7/Treg of spleen.The concentration of tryptophan ( Trp) and kvnurenine ( Kyn) in the liver and the supernatant of FLS were de¬termined by high performance liquid chromatography.Results Compared with model group, ALLO adminis¬ tration significantly increased the body weight of AA rats, reduced the number of joint swelling, improved joint pathological injury,decreased spleen index,inhib¬ited the proliferation of FLS, reduced the number of macrophages in the spleen,decreased Thl7/Treg ratio, inhibited the metabolism of Tip and the production of Kyn in liver and FLS culture supernatant, and de¬creased the liver Kyn/Trp ratio (TOO activity).Con¬clusion ALLO has therapeutic effect on AA rats, which may be related to its regulation of TDO-mediated kyn metabolism pathway.

Objective To quantitatively study the pressure of residual lateral displacement in distal radius AO C3.1 fracture after manual reduction corrected by dynamic airbag pad using finite element analysis and to verify its effectiveness for correcting the residual displacement of fractures. Methods Imageware 13.0, Mimics 15.0 and ANSYS Workbench were used to simulate 1 cm residual lateral displacement after manual reduction of distal radius fracture corrected by dynamic airbag pad. Then the correlation between the distance of residual lateral displacement and the adjustment of dynamic airbag pad pressure were quantitatively analyzed. Results In the case of constant load restrained by airbag ribbon, during the process of pressure adjustment by splint pad, the stress was mainly distributed in the fracture end where the airbag pad was located. About 2.4 kPa pressure was needed to correct 1 mm displacement on radial side, while about 1.3 kPa pressure was needed to correct 1 mm displacement on dorsal side. The dynamic airbag pad was depressurized after the restoration of residual shift. At this time, displacement could be effectively prevented due to the constant load of airbag ribbon and the frictional load at the fracture end. Conclusions In the case of constant load constrained by airbag ribbon, intelligent airbag splint can effectively correct the residual lateral displacement after the manual reduction of the distal radius AO C3.1 fracture and prevent it from being displaced by adjusting pressure of the dynamic airbag pad.

OBJECTIVETo investigate the relationship between acute graft-versus-host disease and graft composition in patients with aplastic anemia(AA) after haploidentical hematopoietic stem cell transplantation.

METHODSFifty-seven cases of AA after haploidentical hematopoietic stem cell transplantation were retrospectively analyzed. All the patients were divided into 2 groups according to whether presence or absence grade Ⅱ-Ⅳ aGVHD, the relationship between aGVHD and graft composition was analyzed by comparing the differences of graft components between the 2 groups.

RESULTSFourteen out of 57 patients had grade Ⅱ-Ⅳ aGVHD and the other 43 did not have grade Ⅱ-Ⅳ aGVHD. The mononuclear cells, CD3, CD4, CD8, NK cells, NKT cells, B cells and Treg cells were not significantly different between the 2 groups (P>0.05), the CD34 cell count in the patients with grade Ⅱ-Ⅳ aGVHD was 3.85（1.73-10.61）×10/kg, which was significantly lower than that without grade Ⅱ-ⅣaGVHD: 6.31（2.98-19.35）×10/kg (P<0.05).

CONCLUSIONSThe incidence of grade Ⅱ-Ⅳ aGVHD may be related with CD34 cell count in AA after haploidentical hematopoietic stem cell transplantation..

OBJECTIVETo investigate the effects of epigallocatechin-3-gallate (EGCG) on proliferation and cell cycle of acute promyelocytic leukemia NB4 cell line and to clarify the molecular mechanism.

METHODSNB4 cells were treated with 0,50,75,100 and 125µmol/L of EGCG for 24, 48, 72 and 96 h, respectively. The proliferation level of NB4 cells was measured by CCK-8 assay. The cell cycle progression of NB4 cells was assayed by flow cytometry. The mRNA expression levels of DNMT1, DNMT3a and DAPK1 were detected by RT-PCR. The methylation status of gene was tested by methylation specific PCR, and the expression level of DAPK1 protein was detected by Western blot.

RESULTSThe proliferation and cell cycle progression of NB4 cells treated with EGCG were inhibited and showed the characteristic of time-dependent and dose-dependent manner. The expression level of DAPK1 and DNMT3a decreased in NB4 cells treated with EGCG. The expression level of DAPK increased in NB4 cells treated with EGCG, while the methylation of DAPK1 gene decreased.

CONCLUSIONEGCG inhibits the proliferation and cell cycle progression of NB4 cells by inhibiting the expression of DNMT1 and DNMT3a and down-regulating the methylation status of DAPK1 gene.

OBJECTIVEGlycogen storage disease type III (GSD III) is an autosomal recessive disease caused by glycogen debranching enzyme (GDE) gene (AGL gene) mutation resulting in hepatomegaly, hypoglycemia, short stature and hyperlipidemia. GSD IIIA, involves both liver and muscle, and accounts for up to 80% of GSD III. The definitive diagnosis depends on either mutation analysis or liver and muscle glycogen debranching enzyme activity tests. This study aimed to establish enzymologic diagnostic method for GSD IIIA firstly in China by detecting muscular GDE activity, glycogen content and structure and to determine the normal range of muscular GDE activity, glycogen content and structure in Chinese children.

METHODMuscle samples were collected from normal controls (male 15, female 20; 12-78 years old), molecularly confirmed GSD III A patients (male 8, female 4, 2-27 years old) and other myopathy patients (male 9, 2-19 years old). Glycogen in the muscle homogenate was degraded into glucose by amyloglucosidase and phosphorylase respectively. The glycogen content and structure were identified by glucose yield determination. The debranching enzyme activity was determined using limit dextrin as substrate. Independent samples Kruskal-Wallis H test, Nemenyi-Wilcoxson-Wilcox test, and Chi-square test were used for statistical analyses by SPSS 11.5.

RESULT(1) GSD III A patients' glycogen content were higher, but G1P/G ratio and GDE activity were lower than those of the other two groups (P < 0.01). In all of the three parameters, there were no significant difference between normal controls and other myopathy patients. (2) The range of normal values: glycogen content 0.31%-0.43%, G1P/G ratio 22.37%- 26.43%, GDE activity 0.234-0.284 micromol/(g. min). (3) Enzymologic diagnostic method had a power similar to that of gene analysis in diagnosis of GSD-IIIA patients. The sensitivity and specificity of enzymologic diagnostic method and mutation detection were 91.7% and 100% respectively.

CONCLUSIONEnzymologic diagnostic method of GSD IIIA was firstly established in China. The range of normal values was determined. This method could be used in diagnosing suspected GSD IIIA patients in the clinic.

Adolescent ; Adult ; Aged ; Biopsy ; Case-Control Studies ; Child ; Child, Preschool ; China ; Female ; Glycogen ; analysis ; Glycogen Debranching Enzyme System ; analysis ; Glycogen Storage Disease Type III ; diagnosis ; enzymology ; pathology ; Humans ; Male ; Middle Aged ; Muscles ; chemistry ; pathology ; Young Adult

Objective To observe the effect of iodine deficiency and hypothyroidism on the protein expressions of calcineurin in the hip-pocampus of pups. Methods Female Wistar rats (n=28) after pregenancy were randomly divided into control group,hypothyroid group and iodine deficient group. According to the dose of propylthiouracil (PTU) in the fed water, hypothyroid group was divided into 5 ppm group and 15 ppm group (7 rats in each group). Totally 5 pups from each group were sacrificed and perfused intracardially in postnatal day (PN) 7,PN14 and PN21. Brains were removed,fixed and sectioned coronally. All sections were observed and analysed for the protein exression of calcineurin by immunohistochemistry in the hippocampus CA1,CA3 and DG regions. Results In PN14 and PN21,protein levels of cal-cineurin in GA1 and CA3 regions of the hippocampus in iodine-deficient and 15 ppm treatment groups were significantly higher than those of the controls (P< 0.05) and in DG region,the contrary was true. In PN7,the positive products were scarely observated in each region and the protein expression was no significantly different in all four groups. Conclusion Iodine deficiency and hypothyroidism may increase the protein expression of calcineurin.

The donkey leukocyte-attenuated vaccine of equine infectious anemia virus (EIAV) was the first lentiviral vaccine that induced solid protection from the infection of virulent strains. To elucidate the mechanism of increased immunogenicity and attenuated virulence of the vaccine, the proviral genomic DNA of an EIAV vaccine strain, EIAV(DLV121) was analyzed and compared with the genome of a parental virulent strain EIAV(DV117). Full length viral genomic DNAs were amplified as two segments by LA-PCR and were cloned. Because of the genomic diversity of retroviral quasispecies, 10 full-length sequences of EIAV(DLV121) and 4 full-length sequences of EIAV(DV117) from randomly picked clones were analyzed. Results showed that the average length of the complete nucleotide sequence of EIAV(DLV121) was 8,236bp and EIAV(DV117) was 8,249bp, with the inter-strain diversity of 2.8%. As for individual genes between the vaccine and virulent strains, the differences in nucleotide sequence of S2, LTR and env were significantly higher than the other genes with the diversity of 4.1%, 3.7% and 3.1%, respectively. Considerable variations in deduced amino acid sequences were found in S2, S3 and env. The diversities were 10.4%, 5.6% and 4.8%, respectively. Furthermore, the LTR of EIAV(DLV121) consisted of at least 5 subtypes grouped by their nucleotide sequences. There were two additional N-linked glycosylation sites in the deduced amino acid sequence of EIAV(DV117) gp90 than that of EIAV(DLV121). Among glycosylation sites in the gp90 of virulent strain, 3 were found unique only in EIAV(DV117), of which 2 were located in the principle neutralizing domain (PND). In addition, there was one EIAV(DLV121) -specific glycosylation site, which was positioned in the PND, too. Taken together, it is clear that greatly increased genomic diversity exists in the EIAV vaccine strain, which provides important information for the further study on biological characters of the Chinese EIAV attenuated vaccine.
Amino Acid Sequence ; Animals ; Base Sequence ; Equidae ; Genome, Viral ; Infectious Anemia Virus, Equine ; chemistry ; genetics ; immunology ; Leukocytes ; immunology ; virology ; Molecular Sequence Data ; Sequence Alignment ; Vaccines, Attenuated ; chemistry ; genetics ; immunology ; Viral Proteins ; chemistry ; genetics ; immunology ; Virulence

Objective A strain of replication deficient recombinant adenvirus encoding fusion glycoprotein (F) of subgroup A human respiratory syncytial virus (RSV) was constructed and the expression of F was identified. Methods The F gone was obtained from pGEM3zf-F with Xho Ⅰ and Hind Ⅲ ,cloned into adenovirase shuttle vector pShutth-CMV,and then the resulting pShutth-CMV/F was transformed into E. coli BJS183/p with pAdeasy-1 to produce pre-adenoviral plasmid encoding F by homologous recombination. This resultant plasmid was linearized by digestion with Pac Ⅰ and transfected into 293 packaging cells to generate FGAd-F. Finally,the expression of F protein was identified by Western Blot analysis. Results FGAd/F was successfully constructed,and the expression of RSV F protein was identified by Western Blot. Conclusion We have obtained a strain of rephcation-defective adenovirus FGAd/F encoding RSV F protein,which can be used further to investigate its protective efficacy against RSV infection in vivo.

OBJECTIVETo investigate the changes in the distribution and chemical states of the hepatic intra- and extra-cellular sodium ion in the rats with severe burns, so as to provide guidance for fluid resuscitation at early postburn stage.

METHODSNineteen adult male Sprague-Dawley (SD) rats were employed in the study and were randomly divided into control (n = 12) and burn (n = 7) groups. The changes in the longitudinal (T1) and transverse (T2) relaxation times of hepatic intra-cellular and extra-cellular sodium in the two groups were studied with 23Na NMR spectroscopy and a shift reagent.

RESULTSAfter infusion of the shift reagent,the extra-cellular sodium content in rat liver decreased by 17%, with obvious increase in fast T2 component (P < 0.01), indicating an increase in the fraction of Na+ binding sites in the extra-cellular space. The characteristics of relaxation of intra-cellular sodium remained unchanged despite a 57% increment in intra-cellular sodium content.

CONCLUSIONThe deficiency of sodium as a permeable molecule might be related to the postburn movement of hypertonic sodium from extra-cellular to intra-cellular space. The results indicated that it is reasonable to administer high concentration of sodium in fluid resuscitation during the first 24 postburn hours.

Animals ; Burns ; metabolism ; physiopathology ; Cations ; metabolism ; Extracellular Space ; metabolism ; Hepatocytes ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Sodium ; metabolism