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 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 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.

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 test the in vitro activity of azithromycin against recent clinical isolates of urogenital Chlamydia trachomatis, and combined activity of azithromycin with moxifloxacin, rifampicin, doxycycline and minocyline. Methods When more than 90% McCoy cells were infected, the 41 strains tested were collected to investigate minimal inhibitory concentrations (MICs) of 5 antimicrobials alone. Checkerboard array was used to calculate the fractional inhibitory concentrations(FICs) and then detected the interactions among the various combinations. Results In vitro, synergism or additivity effect of 51.22%,53.66% and 58.54% strains was found in azithromycin-moxifloxacin, azithromycin-doxycycline and asithromycin-rifampicin combinations, respectively. No difference was observed in all of the combinations ( P ＞0.05). However, antagonism effect of 90.24% strains was observed in azithromycin-minocyline combination. Conclusion This study indicates that the combination of azithromycin with moxifloxacin, doxycycline or rifampicin is more effective against Chlamydia trachomatis than individual antimicrobials. Therefore, these antimicrobials combinations might be recommended against Chlamydia trachomatis recurrent or persisitent infection. However, the combination of azithromycin with minocyline exhibited a markedly antagonism activity against Chlamydia trachomatis. Combined sensitivity test to a certain extent can compensate for some shortcomings of individual susceptibility test.

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 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 compare the infectivity of several transformed Chlamydia trachomatis (Ct) mouse pneumonitis (Mopn) strains to host cells, and to identify cell invasion-related virulence genes in Chlamydial plasmids. Methods Several Ct strains, including wild-type Ct Mopn strain(WT strain), plasmid-free Ct strain(CMUT3 strain), Ct Mopn strain transformed with the shuttle vector carrying pGFP and the complete C. muridarum (CM) plasmid (pGFP::CM strain)and Ct Mopn strains transformed with shuttle vectors carrying pGFP and mutant CM plasmids with in-frame deletions of Pgp3, 4, 5 or 7 (pGFP::CM△Pgp3, 4, 5, 7 strains), were cultured, amplified and collected. After the concentrations of Ct were determined, each of these strains was divided into four groups to be inoculated at a same amount(1.5 × 104 inclusion forming units(IFU)) followed by four different treatments respectively: centrifugalization +DEAE group treated with centrifugalization followed by ion-exchange chromatography on diethylaminoethyl(DEAE)-cellulose columns, centrifugalization group treated with centrifugalization only, DEAE group treated with chromatography on DEAE-cellulose columns only, control group receiving no treatment. After additional culture for 20 - 24 hours, indirect immunofluorescence assay was performed to count the number of chlamydial inclusions. At 20, 40 and 60 hours after infection, the growth rate and area of chlamydial plaques were assessed after three continuous passages. Lugol′s iodine staining was conducted to observe glycogen synthesis in bacterial inclusions. Results The inclusion number in the centrifugalization + DEAE group, centrifugalization group, DEAE group and control group was 10.20 ± 1.30, 6.80 ± 0.44, 3.00 ± 1.22 and 0.80 ± 0.45 respectively for the pGFP::CM△Pgp4 strain, 6.40 ± 0.89, 3.80 ± 0.83, 1.60 ± 0.89 and 0.60 ± 0.54 respectively for the CMUT3 strain. Under same experiment conditions, the pGFP::CM△Pgp4 strain and CMUT3 strain showed similar infectivity, and formed less inclusions compared with the other Ct strains (all P < 0.01). The number of inclusions formed by the same Ct strains were significantly different among the 4 groups(F = 845.310, P <0.01), and were highest in the centrifugalization + DEAE group for all the strains. The pGFP::CM△Pgp4 strain showed significantly lower growth rate and area of plaques with an abnormality in glycogen accumulation compared with the other strains at 20, 40 and 60 hours after infection. Conclusion The plasmid-encoding gene Pgp4 may be a cell invasion-associated virulence gene in chlamydial plasmids.