Objective To explore the role of a new apoptosis related gene TFAR 19 in the pathogenesis of systemic lupus erythematosus (SLE) and the relationship between TFAR 19 and SLE.Methods ELISA was used to test if there is relation between TFAR 19 and SLE.Results It was found that in active SLE patients there was higher titer of TFAR 19 antibody than that in stability patients and normal controls.No significant difference was seen between stability patients and normal controls.Conclusion It is first put forward that TFAR 19 may be related with SLE pathogenesis and disease activity.

Objective: To establish an efficient expression system for human truncated insulin like growth factor 1 〔Des(1 3)IGF1〕as fusion protein in Escherichia coli(E.coli). Methods: The cDNA of Des(1 3)IGF1 was cloned into an fusion protein expression plasmid, pMTY4, using gene recombinant technique. The protein was purified by ion exchange chromatography and identified by SDS polyacrylamide gel electrophoresis, radioimmunoassay(RIA), N terminal amino acid sequence and biological activity. Results: A prokaryotic expression vector was constructed and the fusion protein containing MS2 polymerase fragment, thrombin recognition site and human Des(1 3)IGF1 was expressed in E.coli at high level. It was showed that the purified recombinant Des(1 3)IGF1 released from the fusion protein after digestion with thrombin was identical to the native Des(1 3)IGF1.Conclusion:This is an effective method for obtaining human recombinant Des(1 3)IGF1 and it is very important for further study of Des(1 3)IGF1.

Objective: in order to provide rapid and reliable method. Methods: Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E　coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis. Results:Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis. Conclusion: We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.

Objective: To study the expression and localization of apoptosis-related protein TFAR19 in TF-1 cells undergoing apoptosis. Methods: Using monoclonal antibody against TFAR19, the expression level and cell localization of TFAR19 were examined by fluorescence microscope, confocal laser scan microscope(CLSM) and flow cytometry. Simultaneously, we also analyzed the relationship of TFAR19 protein with phosphatidylserine (PS) externalization and cell nuclear DNA fragmentation. Results: The level of TFAR19 proteins expressed in TF-1 cells treated with GM-CSF withdrawal was significantly increased compared with normal TF-1 cells, then translocated rapidly from cytoplasm to the nucleus of cells. Appearance of TFAR19 in the nucleus of apoptotic cells preceded the detection of PS externalization and DNA fragmentation. Conclusion: Nuclear translocation of TFAR19 protein is one of the earliest events of cell apoptotic process. These data provided a new clue to further approach to the biological function of TFAR19 and study of cell apoptosis.

AIM: To clarify the role of new apoptosis-related gene, TF-1 cell apoptosis-related gene 19(TFAR19), in the pathogenesis of systemic lupus erythematosis (SLE) and the relationship between TFAR19 and SLE. METHODS: DNA Ladder detection, Western blotting, immunological fluorescence method, ELISA and so on were used to test if ultraviolet B(UVB) could induce HaCaT cell apoptosis and TFAR19 expression. RESULTS: HaCaT cell apoptosis could be detected after 24 hours of 30 mj/cm 2 UVB irradiation. Also, we found that in active SLE patients, the TFAR19 antibody was increased, but not significant compared to the normal control. CONCLUSION: TFAR 19 is involved in the process of UVB induced ketatinocyte line HaCaT apoptosis and SLE pathogenesis.

OBJECTIVETo explore the effects of a novel human chemokine-like factor 1 (CKLF1) on stem cell/progenitor cells.

METHODSHuman bone marrow mononuclear cells were separated by Ficoll (1.077 g/ml), and cultured in suspension and semisolid colony culture. The effects of CKLF1 on CFU-GM and CFU-Mix were observed.

RESULTSThe number of CFU-GM was significantly increased in 10 groups by addition of CKLF1 plasmid supernatant. The mean value was 234.81 +/- 98.71/1 x 10(5) cells, 2.42 fold of control group (P < 0.05). The mean value of CFU-Mix in groups of negative control, CKLF1, PHA, GM-CSF and G-CSF were 82.00 +/- 20.25, 105.76 +/- 36.70, 146.63 +/- 27.09, 143.33 +/- 60.23 per 1 x 10(5) cells, respectively, no statistical differences were seen between control and CKLF1 groups. The CD(34)(+) cells were detected by FACS. The average percentage in the groups of normal control, CKLF1, PHA and GM-CSF were (0.75 +/- 0.62)%, (1.32 +/- 0.87)%, (0.15 +/- 0.02)%, and (0.29 +/- 0.23)%, respectively. Compared with control, no significant differences were seen between each group (P > 0.05).

CONCLUSIONNovel chemokine-like factor 1 can increase the proliferation of CFU-GM, which indicated that CKLF1 could increase the proliferation of human bone marrow hematopoietic progenitor cells and colony formation.

Adult ; Cells, Cultured ; Chemokines ; pharmacology ; Hematopoietic Stem Cells ; cytology ; drug effects ; Humans ; MARVEL Domain-Containing Proteins

In recent years, with the widespread clinical application of moxibustion, the clinical trials on moxibustion research are also increasing. However, the establishment of negative control of moxibustion, also called as mimicking moxibustion, has become a major challenge in the randomized controlled trials (RCTs) . In this review, we summarized the establishment methods of mimicking moxibustion in RCTs at home and abroad, and evaluated and elaborated the design and application methods of mimicking moxibustion models from thermal effect, light radiation effect and smoke effect that generated by moxibustion, in order to provide insights for the establishment of an ideal and feasible mimicking moxibustion model in future RCTs of moxibustion.

ObjectiveTo investigate the effects of thyme herbal tea （BLX） on the proliferation of human coronavirus OC43 （HCoV-OC43） in vitro and in vivo. MethodThe chemical composition of BLX was analyzed by UPLC-MS. The cytotoxicity of BLX in HRT-18 cells and the effect of BLX treatment on the proliferation of HCoV-OC43 in cells were analyzed. Copies of viral gene were detected by real-time PCR. The effect of BLX treatment on the life cycle of HCoV-OC43 was detected by time-of-addition assay. The maximum tolerated dose of BLX and the influences of BLX on the body weight and survival time of suckling mice infected with HCoV-OC43 were determined. The expression of viral protein in the brain and lung tissue was analyzed by immunohistochemistry. ResultThere were 11 chemical components identified in BLX by UPLC-MS. BLX showed the 50% cytotoxic concentration （CC₅₀） of （13 859.56±319） mg·L^-1， the median inhibitory concentration （IC₅₀） of （1 439.09±200） mg·L^-1， and the selection index of 8.26-11.44 for HCoV-OC43 in HRT-18 cells. Compared with the cells infected with HCoV-OC43， BLX at the concentrations of 1 500， 1 000， 500 mg·L^-1 inhibited the proliferation of this virus （P<0.05， P<0.01）. BLX exhibited antiviral effect in the early stage of virus infection， and the inhibition role in the attachment stage was more significant than that in the entry stage （P<0.05）. In the suckling mice infected with HCoV-OC43， BLX at 1200 and 600 mg·kg^-1·d^-1 alleviated the symptoms， prolonged the survival period， reduced the death rate， and down-regulated the mRNA level of nucleocapsid protein in the mice. Moreover， BLX at 1 200 mg·kg^-1·d^-1 down-regulated the expression of nucleocapsid protein in the brain （P<0.01） and the lung （P<0.01）. ConclusionBLX contained multiple antiviral ingredients. It inhibited the proliferation of HCoV-OC43 both in vitro and in vivo by interference with viral attachment. This study provides theoretical reference for the treatment of acute respiratory tract infection with HCoV-OC43 and for further development and application of BLX.