The levels of low-density lipoprotein(LDL)glycation from control group,diabetic HbA1C < 7.0%,and HbA1C>7.0% groups were(17.7±2.31),(34.29±5.73),and(48.79±7.82)Glycogroups/LDL by fluorimetry.The LDL binding to its receptor in three groups were(37.65±5.20),(27.36±4.34),and(15.07± 2.23)ng/mg cell protein measured by enzyme-linked immunoreceptor assay.The glycated levels in two diabetic groups were higher than that in control group,and higher in HbA1C>7.0% group than in HbA1C<7.0% group(all P< 0.01).The results of LDL binding capacity to its receptor were just the opposite.

Objective To express the recombinant mouse histidyl-tRNA synthetase (HARS) and maltose binding protein (MBP) gene in Escherichia coli and obtain the purified protein which possesses antigen specificity.Methods Total RNA was extracted from the myocytes of C57BL/6 mouse and reversely transcripted to cDNA.The gene of N-terminal origin of 591 base pairs was amplified,then cloned into pMALc-5e vector.The recombinant plasmid was transformed into Rosetta-gami B,then IPTG was used to induce the expression of HARS-MBP.Fusion protein was purified by affinity chromatograph.The molecular weight (MW) of HARS-MBP was roughly determined by SDS-PAGE.The antigen specificity was identified by Western blotting using anti-Jo-1 serum from patients,commercial anti-HARS and anti-MBP antibodies.Results The recombinant HARS-MBP protein gene was efficiently expressed in Escherichia coli,and the MW was consistent with predicted MW of 66 000.The fusion protein was specifically combined with its antibody.Conclusion The HARS-MBP fusion protein could be efficiently and steadily synthesized in Escherichia coli,which shows satisfactory antigen specificity and provides the key requirement for making a deep study of HARS in the pathogenesis of idiopathic inflammatory myopathy(IIM) and animal modeling of IIM.

To investigate the effects of Ligustrazine on histogenesis of bone marrow in the early phase of hematopoietic reconstruction in bone marrow transplantation (BMT) mice. The syngeneic BMT mice model was established. The syngeneic BMT mice were orally given 2 mg Ligustrazine twice a day. 1, 3, 5, 7, 10, 15 and 21 day(s) after BMT, peripheral blood granulocytes and bone marrow nucleated cells (BMNC) were counted and the diameter of central vein and the area of micro-vessel in femur were measured. The effect of Ligustrazine on hematopoietic stem cells was observed by colony forming unit of spleen (CFU-S). The effect of Ligustrazine on hemopoietic progenitors was studied by observing the number of progenitors of Granulocytes/Macrophage on day 10 and day 20 after BMT. In Ligustrazine-treated group, the diameter of center veins and the area of micro-vessel of femur were all significantly less than the control group 7, 10, 15, 21 days after BMT (P < 0.01). In addition, Ligustrazine significantly increased the number of CFU-S on day 10 and the number of CFU-GM on day 10, 20 after BMT. These results indicate that Ligustrazine can accelerate the histogenesis of hemopoietic bone marrow, which may be one mechanism by which Ligustrazine promotes hematopoietic reconstitution after BMT.
*Bone Marrow Transplantation ; Hematopoiesis/*drug effects ; Hematopoietic Stem Cells/*drug effects ; Mice, Inbred BALB C ; Pyrazines/*pharmacology ; Time Factors

Objective This study was focused on the association between neutrophil extracellular traps (NETs) and interstitial lung disease (ILD) in patients with dermatomyositis (DM).Methods Thirty six patients who satisfied the Bohan & Peter criteria for DM were recruited to this study,among whom 19 were complicated with ILD.Forty seven age and sex matched healthy Chinese volunteers were selected to be control subjects.The plasma samples of these patients were tested for the formation and degradation of NETs.Results DM plasma induced more NETs formation than control plasma did [(246±93) RFUs vs (192±53) RFUs,P=0.002].Compared to control,DM plasma exhibited a signficantry decreased ability to degrade NETs.Further mere,compared with DM patients without ILD (DMNL),DM patients with ILD (DML) could not degrade NETs completely [(83±13)％ vs (59±21)％,P＜0.01].All four DM patients with subacute ILD exhibited a significantly lower ability to degrade NETs than patients with chronic or asymptomatic ILD [(36±14)％ vs (65±19)％,P=0.0139].Conclusion These data show that more NETs formation is induced by plasma and DML fails to completely degrade NETs.These suggest that NETs may play a role in the pathogenesis of DM and DM-associated ILD.

Objective To establish a new murine model of experimental autoimmune myositis by immunizing with MYBPC2 protein. Methods The purified Myosin-binding protein C, fast type (MYBPC2) was emulsified with complete Freundˊs adjuvant, then C57BL/6 mice were immunized by multi-point subcutaneous injection (0, 7 days), and intraperitoneal injection of pertussis toxin 2 μg simultaneously. The pathological changes of mice with different immunizing dose at the preconceived time were ex-plored. Mean-while, mice were immunized with 600 μg each time, and the muscle endurance was tested on the 21th day. The expression of major histocompatibility complex (MHC) class-Ⅰ and the surface biomarkers of the inflammatory cells in muscle tissues were observed. Mann-Whitney U test was used for statistical analysis. Results ① With the increase of immunizing dosage, muscle damage and inflammation tended to be more serious. On the 21th and 28th day, muscle lesions were most significant. Muscle fiber degeneration and necrosis and inflammatory cell infiltration could be seen in the experimental group. ② Compared with the control group, muscle endurance of mice in the experimental group decreased significantly [(6.1 ±1.3) min versus (9.2±1.6) min, U=2.00, P=0.017]. The MHC class-Ⅰ on the muscle fiber surface of the experimental group was positive, scattered infiltration of CD4 +, CD8+ T ly-mphocytes and CD68 + macrophages between muscle fibers and around the vascular areas could be observed, and CD20+B lymphocytes mainly distributed in the area around the blood vessels, nevertheless rarely seen between muscle fibers. Conclusion Exper-imental autoimmune myositis models of mice have been successfully induced by immunizing with MYBPC2 in China for the first time, and similar clinical and pathological features of human polymyositis could be observed. This new model can be used for studying the pathogenesis of autoimmune myositis.

Objective To analyze CYP17A1 gene mutation in a patient with 46,XY disordered sex development and to explore the possible influence on the phenotype of the patient.Methods Eight exons of CYP17AI gene in the patient and her parents were amplified and directly sequenced.In order to construct Mini-gene system,PCR fragments containing wildtype and mutant splicing sites were inserted in expression vector,and then transfected into cells.RT-PCR was used to observe the influence of splicing site mutation.Wildtype and aberrant splicing CYP17A1 cDNA expression plasmids were constructed and transfected into cells respectively,and CYP17A1 enzyme activity was tested in vitro.Results Mutation analysis revealed compound heterozygous CYP17A1 mutations,with Y329fs in one allele and a synonymous substitution( c.1263G＞A:GCG＞GCA) in another allele.In vitro analysis showed that the synonymous substitution induced a novel splicing site,which resulted in aberrant splicing of CYP17A1 mRNA and lacked six or seven amino acids after 415 in splicing product.In vitro transfection and enzyme activity experiment showed that the aberrant splicing product abolished the enzyme activity completely.However,this mutation did not completely influence splicing.The patient also had a part of normal splicing product,which was a coincidence to the phenotype of the patient.Conclusion This is the first description of an exonic splicing mutation in CYP17A1 relevant to the 17ot-hydroxylase deficiency phenotype.The functional study of the aberrant splicing variant has been initiated.

To investigate the effects of Ligustrazine on histogenesis of bone marrow in the early phase of hematopoietic reconstruction in bone marrow transplantation (BMT) mice. The syngeneic BMT mice model was established. The syngeneic BMT mice were orally given 2 mg Ligustrazine twice a day. 1, 3, 5, 7, 10, 15 and 21 day(s) after BMT, peripheral blood granulocytes and bone marrow nucleated cells (BMNC) were counted and the diameter of central vein and the area of micro-vessel in femur were measured. The effect of Ligustrazine on hematopoietic stem cells was observed by colony forming unit of spleen (CFU-S). The effect of Ligustrazine on hemopoietic progenitors was studied by observing the number of progenitors of Granulocytes/Macrophage on day 10 and day 20 after BMT. In Ligustrazine-treated group, the diameter of center veins and the area of micro-vessel of femur were all significantly less than the control group 7, 10, 15, 21 days after BMT (P < 0.01). In addition, Ligustrazine significantly increased the number of CFU-S on day 10 and the number of CFU-GM on day 10, 20 after BMT. These results indicate that Ligustrazine can accelerate the histogenesis of hemopoietic bone marrow, which may be one mechanism by which Ligustrazine promotes hematopoietic reconstitution after BMT.
Animals ; Bone Marrow Transplantation ; Female ; Hematopoiesis ; drug effects ; Hematopoietic Stem Cells ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Pyrazines ; pharmacology ; Time Factors

OBJECTIVE:To reduce the incidence of irrational medical orders for parenteral nutrition,and promote the rational use of parenteral nutrition. METHODS:The medical orders for parenteral nutrition of the first quarter of 2016 in general surgery de-partment of our hospital were collected,and the number and types of its irrational medical orders were summed up. Then FO-CUS-PDCA(Find-organize-clarify-understand-select-plan-do-check-act)cycle management was adopted to analyze and improve the existing problems in issuing medical orders for parenteral nutrition. The improved(the third quarter of 2016)medical orders for par-enteral nutrition were collected,the number and types of its irrational medical orders were summed up,and management effect was evaluated. RESULTS:Establishing nutrition support group,strengthening the training and communication of medical staff,adding prescription evaluation module for parenteral nutrition in hospital information system and a number of measures had made the inci-dence of irrational medical orders for parenteral nutrition in general surgery department declined from 48.25%(1433/2970)before improvement to 5.67%(120/2118)after improvement. The incidences of cation excess,inappropriate selection of drugs and inap-propriate compatibility in irrational types were 0. CONCLUSIONS:FOCUS-PDCA cycle management can reduce the irrational rate of medical orders for parenteral nutrition and promote the rational use of parenteral nutrition in hospital.