Objective To study the effctiveness and safety of arsenic trioxide(As2O3)combined with Homoharringtoninum in the chronic myelocytic leukmia-chonic phase(CML-CP).Methods Seven patients were treated with As2O3 10mg per day for 2~3 week and with Homoharringtoninum 3~4mg per day for 1~2.Results All patients were clinic remission,of which there were 4 complete remission who were all initial therapy,and 3 partial remission,2 of who were initial therapy and 1 was resume threapy.4 patients of CR were treated with second strengthen therapy and continued hematologic CR.The main side effects were grade 3 hematologic toxicity and light liver damage,and no significant nausea,emesis,diarrhea or mucositis.Conclusion As2O3 combined with Homoharringtonium in the CML-CP is a safe and effctive regimen.

Objective To compare the clinical efficacy and safety of two different conditioning regimens in nonmyeloablative autologous peripheral blood stem cell transplantation (NAST) for the treatment of systemic lupus erythematosus (SLE).Methods Different conditioning regimens were used in two groups:cytarabin combined cyclophosphamide in group 1 and ATG combined cyclophosphamide in group 2.Different recovery time of leucocytes,neutrophils and platelets in the two groups were compared.Statistical analysis were carried out by paired t-test.Results The mean time for peripheral leucocytes reaching 1.0×109/L,neutrophils getting up to 0.5×109/L,platelet raising to 100×l09/L and hemoglobin rising to 120 g/L in group 1 were [(7.2±1.3),(8.0±1.5),(10.5±1.4),(22.1±2.3)days] and [(10.4±2.1),(12.0±1.9),(19.3±2.1),(28.1± 2.4)] days in group 2.The difference was statistically significant (P＜0.01).CD4+ cell count and the ratio of CD4+/CD8+ of pre- and pro-NAST was changed.No significant differences were observed in the two groups.Conclusion For the sake of safety and hematopoietic reconstitution,we recommend cytarabin combined cyclophosphamide as the preferred conditioning regimen.

Objective To investigate the relationship between serum visfatin level and coronary artery stenosis.Methods Based on coronary angiography,85 patients were diagnosed as coronary heart disease.According to oral glucose tolerance test,these patients were divided into 3 groups,34 patients with normal glucose tolerance(CHD group),25 with impaired glucose regulation(CIG group),and 26 with type 2 diabetes mellitus(CDM group).30 non-comary heart disease subjects with normal glucose tolerance were selected as the control group(CON group),and they underwent coronary CT angiography scan and were confirmed coronary disease-free.Blood pressure,body mass index(BMI),waist circumference(WC),waist hip ratio(WHR),fasting plasma glucose(FPG),blood lipid analysis,fasting insulin,and HbA1C were determined.Serum visfatin concentration was assayed and the status of coronary artery was assessed.Coronary artery stenosis was screened by coronary interventional angiography and assessed by Gemini scoring system in CHD,CIG,and CDM groups.Results Compared with control group,serum visfatin in CHD,CIG,and CDM groups were significantly higher(all P<0.05).Compared with CHD group,serum visfatin in CIG and CDM groups were significantly higher(all P<0.05).The correlation analysis showed that serum visfatin level was positively correlated with the involved branches of coronary arteries(r=0.807,P<0.01),serum visfatin level was positively related with Gensini coronary artery score(r=0.669,P<0.01).Visfatin was also positively correlated with WC,WHR,triglyceride(TG,r=0.200,P=0.032,r=0.185,P=0.047,r=0.321,P<0.01),while high density lipoprotein cholesterol(HDL-C)was negatively correlated with visfatin(r=-0.354,P<0.01).Multiple regression analysis showed that senlm hvel of TG and WC were the main influencing factors of visfatin.Conclusion (1)The level of serum visfatin may reflect the severity of coronary artery stenosis,detection of visfatin helps to make early diagnosis of CHD.(2)The raised serum level of visfatin in comary heart disease patients with imparied glucose tolerance is consistent with clinical evidence that diabetic patients have more severe coronary diseases.(3)WC and serum TG are main inilucencing factors,suggesting that visfatin is correlated with abdominal obesity.

Objective To investigate the long-term efficacy of nonmyeloablative autologous peripheral blood stem cell transplantation(NAST) to cure refractory autoimmune disease(AD).MethodLong-term follow up of four cases of AD patients with NAST were summarized.The pretreatment regimen was intravenous injection of cytarabin (200 mg· kg-1· d-1 ) and cyclophosphamide (40 mg· kg-1· d-1).The therapeutic effect was evaluated by the change of symptoms and signs and long term complications.Changes of immune function were detected by flow-cytometry.ResultsFive cases of patients had been successfully engrafted.The average time for peripheral leucocytes count to reach 4.0×109/L was 12 days.It needed 10 days for platelets to return to 100×109/L and 22 days for hemoglobin to 120 g/L.Apparent remission of symptoms and signs was observed after transplantation.Lymphocyte subtypes analysis pre- and post- NAST showed that count of CD4+ and the ratio of CD4 +/CD8 + was returned to normal.One patient gave birth to a healthy baby four years after transplantation.Three female patients returned tonormal life. Conclusions Compared with classical myeloablative stem cell transplantation,NAST has a rapid hematopoietic recovery and good long-term therapeutic effect in AD.The quality of life in AD patients treated with NAST is higher than those treated with myeloablative hematopoietic stem cell transplantation.

Objective To detect Chlamydia trachomatis phage Vp1 gene in clinical swab specimens and anti-Vp1 antibodies in serum specimens.MethodsCervical and urethral swab as well as serum specimens were collected from attendees to the sexually transmitted disease(STD) clinic in the Tianjin Institute of STD,during March 2008 to March 2011.PCR was conducted to detect chlamydial phage Vp1 gene in swab samples,enzyme linked immunosorbent assay(ELISA) and Western blot to detect anti-Vp1 antibody in sera.The swab specimens positive for Vp1 gene were subjected to cell culture followed by the detection of Vp1 protein with an immunofluorescence-based method.ResultsTotally,36 out of 1542 swab specimens turned out to be positive for Vp1 gene,and 23 out of 453 serum specimens for anti-Vp1 antibody.No positive results were obtained in the Vp1 gene-positive swab specimens by cell culture and immunofluorescence-based assay.ConclusionThe Vp1 gene of Chlamydial trachomatis phage and anti-Vp1 antibody are successfully detected from clinical swab and serum specimens respectively.

Objective To evaluate the significance of platelet activation and the expression of inter-leukin (IL)-1β in patients with RA. Methods The activation of platelets and the expression of IL-1β in pla-telets in 50 RA patients(22 high-active, 28 mediate/low active ) and 30 normal controls were determined us-ing flow cytometry. Meanwhile, inflammatory indicators such as erythrocyte sedimentation(ESR), C-reactive protein (CRP) and DAS28 were also recorded. T test and correlation analysis were performed. Results The platelet activation in RA group(19.2±4.8) was higher than the control group(9.0±2.9)(t=10.5, P=0.001). The expression of IL-1β in platelets in RA group(41±11) was higher than control group(21±8)(t=9.01, P =0.000) .The platelet activation in high-active RA group(22 ±4) was higher than mediate/low active RA group(17 ±4)(t =3.96,P =0.001). The expression of IL-1β in platelets in high-active RA group(45 ±10) was higher than mediate/low active RA group (38 ±10)(t =2.329,P =0.024). The expression of IL-1β in platelets in RA group was positively correlated with the level of ESR、CRP and DAS28 (r value and P value were 0.576, 0.578, 0.618 and 0.000, 0.000, 0.000 respectively). Conclusion The platelets of patients with RA are activated and may suggest that IL-1β, which may associate with disease activity. Our research suggest that platelet may play a role in the inflammatory process of RA by secreting IL-1β.

Objective To get phagc's capsid Vp3 gene and protein of guinea pig inclusion conjunctivitis (GPIC) chlamydia. Methods The genome DNA was extracted from the φCPG1 phage.The full sequence of Vp3 gene was amplified by polymerase chain reaction (PCR) from the above genome DNA. The Vp3 gene was digested by restriction endonuclease and then inserted into prokaryotic plasmid vector pET30a (+). The recombinant plasmid was transformed into E. coil BL21, and was identified by restriction endonuclease, PCR and sequencing. The E. coil BL21 with expected recombinant plasmid was induced and the expressed recombinant Vp3 protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, then purified by agarose gel. Results The recombinant gene was sequenced and proved to be 447 bp which was consistent with the φCPG1 Vp3 gene sequence in GenBank. A 25 000 capsid protein was expressed and confirmed by SDS-PAGE and Western blot. The purified protein was obtained. Conclusion The capsid Vp3 protein of φCPG1 is successfully expressed and purified, which is helpful for the further study on its mechanism and clinical applications.

Objective To test the in vitro acitivity of azithromycin, minocyline, moxifloxacin, doxycycline and rifampicin alone or in combination against three standard strains of urogenital Chlamydia trachomatis,i.e. D-UW-5/Cx, E-UW-5/Cx and G-UW-5/Cx. Methods Three standard strains of C. trachomatis were inoculated into McCoy cells, and used to susceptibility testing after more than 90% McCoy cells were infected.Microdilution method was applied to determine the activity of the 5 antimicrobial agents alone, and checkerboard array to determine that in combination. Results The in vitro minimal inhibitory concentrations (MICs)were 0.25 mg/L for azithromycin, 0.06 mg/L for moxifloxacin, 0.063 - 0.25 mg/L for doxycycline, 0.032 -0.064 mg/L for minocyline, 0.008 mg/L for rifampicin. And, the fractional inhibitory concentration index was constantly 0.75 for the combination of azithromycin with moxifloxacin, doxycycline and rifampicin, 2.5, 2.83and 4 for the combination of minocyline with azithromycin, moxilloxacin and rifampicin, respectively. Conclusions As far as the activity against C. trachomatis is concerned, there is a synergism for the combination of azithromycin with moxifloxacin, doxycycline and rifampicin, but antagonism for the combination of minocyline with azithromycin, moxifloxacin and rifampicin.

Objective To express recombinant Vp1 protein of chlamydiaphage φCPG1, prepare monoclonal antibody against Vp1 protein and utilize it to screen clinical isolates of Chlamydia trachomatis. Methods The Vp1 protein was obtained by prokaryotic expression, and monoclonal antibody against this protein was prepared by hybridoma technique. ELISA and Western blot were used to identify monoclonal antibodies. Then,the monoclonal antibody was prepared in quantity by injecting hybridoma cells into the abdominal cavity of BALB/C mice, and purified by using protein G affinity chromatography. Clinical isolates of Chlamydia trachomatis were screened for the chlamydiaphage by immumofluorescence assay using the prepared monoclonal antibody.Results Purified Vp1 protein was obtained. The monoclonal antibody against Vp1 protein was gained after 3times of sub-clone and consistently identified as IgG1. Three hybridoma cell strains that stably secreted monoclonal antibody were generated. Chromosome analysis of hybridoma cells showed that the mean number of chromosome was 96, most of them were telocentric and a few were submetacentric. The titer of purified monoclonal antibody was more than 1: 12 800. Twenty clinical isolates were screened by using the monoclonal antibody and no positive results were obtained. Conclusions The monoclonal antibody against Vp1 protein of chlamydiaphage φCPG1 is successfully prepared, while no chlamydiaphage is detected by immumofluorescence assay using the prepared antibody in 20 clinical isolates of Ct.

Objective To investigate specific immune responses in mice induced by recombinant major outer membrane protein(rMOMP)of C.trachomatis serovaf E.Methods Thirty-six female BALB/cmice aged 3 to 4 weeks Were divided into three groups.i.e.,adjuvant group vaccinated、with purified rMOMP and Freund's adjutant,solitary group vaccinated with rMOMP only and control group vaccinated with phosphate buffered saline(PBS).All the mice were intramuscularly vaccinated on week 0,2 and 4.Blood samples and vaginal washes were obtained from these mice on week 6,then,mice were challenged with elementary body(EB)of C.trachomatis serovar E at the footpad followed by the observation of delayed hypersensitivity.On week 7.mice were genitally infected with C.trachomatis EB;one week later,blood samples and vaginal washes were obtained again;six weeks later,spleen lymphocytes were isolated from the mice and stimulated bv C.trachomatis or ConA followed by the detection of cell proliferation with MTT assay.In vitro neutralization assay was also performed.ELISA was used to determine the titers of Chlamydia-specific IgO antibody in sera and IgA antibody in vaginal washes,as well as the level of IFN-γ in culture supernatant of lymphocytes and sefa of mice.Vaginal swabs were collected after genital challenge and subjected to C.trachomatis culture.Results The absorbance at 405 ms of Chlamydia-specific IgG antibody and proliferation index of lymphocytes were 0.641±0.059 and 5.085±1.291.respectively,in mice immunized with rMOMP and Frennd's adjuvant.significantly higher than those in mice immunized with rMOMP only(0.424±0.015 and 3.123 ±0.840.both P<0.05).The thickness of right hind footpad increased by 0.324±0.054 mm and 0.272±0.064 mm,respectively,in solitary group and adjuvant group,respectively,with significant difference between the two groups(P<0.05).A significant increase was also observed in the adjuvant group compared with the control group in the above three parameters(all P<0.01).Conclusion The rMOMP of C.trachomatis could efficiently induce Chlamydia-specific humoml and cellular immune responses in mice.