Objective To observe the curative effects and toxicity of low-dose bortezomib plus thalidomide and chemotherapy in treatment of multiple myeloma. Methods 35 patients with initial, refractory or relapsed MM received at least two cycles of treatment with bortezomib at 1.1 mg/m2 intravenously on days 0,3, 7, and 10, and by daily oral thalidomide escalated from 50mg to 150 mg and chemotherapy. The chemotherapy regimens included MP, VAD and AD regimen which was chosen according to the status of patients. Results After a median follow-up of 20 months, the overall response rate was 82.8 %, complete remission (CR)48.6 %, very good partial remission (VEPR) 17.1%, and partial remission 17.1%. The 3-year PFS and OS were 60.92 % and 72.41% separately. ORR and OS were same in initial and refractory or relapsed MM patients. Grade 3 or 4 adverse events including debility (3/35), nausing and vomiting (8/35), constipation (4/35), peripheral sensory neuropathy (3/35), neutropenia (10/35) and thrombocytopenia (12 %) were observed.Conclusion The regimen of low-dose bortezomib plus thalidomide and chemotherapy is a highly effective and safety regimen for MM patients. The maintenane therapy with thalidomide may prolong PFS.

Objective To study the effect of astragaloside on proliferation and apoptosis in human keloid fibroblasts.Methods The human keloid fibroblast ceils were treated with different concentration of astragaloside(10、20、40 ng/mL).Cell proliferation was detected by MTT,the gene expreesion levels and protein levels of apoptosis-related proteins,survivin,p53 and Bcl-2.were determined by real-time PCR and Western blot,respectively.Results Comparecl with control group(treated with 0 ng/mL astragaloside),the absorbance values (A490 nm) of each concentration group were significantly reduced,which suggest that the proliferation of all keloid fibroblast were markably inhibited in a dose-dependent way (P＜0.05).The gene expreesion levels and protein levels of apoptosis-related proteins,survivin、Bcl-2 were largely suppressed and P53 werelargely promoted in a dose-dependent.Conclusion The keloid fibroblasts cells proliferation and apoptosis could be regulated by astragaloside.

Objective To construct eukaryotic expression recombinant plasmid containing human granulysin(GLS) and investigate the effect of GLS expression in macrophage RAW264.7 cells on the bactericidal activity against intracellular Mycobacterium tuberculosis.Methods GLS gene was amplified by nested-polymerase chain reaction(PCR) from human cytotoxicity T lymphocyte(CTL) activated by allogenic antigen,and inserted into pBudCE4.1 vector to construct recombinant plasmid.Subsequently,the plasmid was transfccted into RAW264.7 cells which were infected with Mycobacterium tuberculosis H37Rv.The expression of GLS was detected by nested-PCR and immunocytochemistry method.The RAW264.7 cells were lysed after transfected for 96 h,then acidfast stained,cultivated and colony count were done to determine the intraeellular bactericidal activity of GLS.The data were analyzed by t or t' test.Results The pBudCE4.1/GLS eukaryotic expression recombinant plasmid was successfully constructed.The transcriptional and translational expressions of target gene GLS were detected in RAW264.7 cells which were infected with Mycobacterium tuberculosis H37Rv.The bacterial load in macrophages of phorbol myristate acetate(PMA)+pBudCE4.1/GLS group,PMA+pBudCEA.1 group and non-activated group were 1.44±1.25,3.16±0.20 and 3.59±0.21,respectively.The differences between groups were all significant (t=2.403,t=2.854,both P＜0.05).Conclusion Eukaryotic expression recombinant plasmid carrying human GLS gene expressed in macrophages has strong bactericidal activity against intracellular mycobacteria,which provide information for the further study on therapeutic vaccine against Mycobacterium tuberculosis.

Objective To observe the clinical effect on sleep improvement about acupoint application in treating the chronic insomnia of Liver-stagnation and Spleen-deficiency Syndrome.Methods A total of 68 patients with chronic insomnia with Liver-stagnation and Spleen-deficiency Syndrome were randomly and blindly divided into treatment group and control group by registration order. In the treatment process, 1 was eliminated and 33 were completed in the treatment group, 2 were eliminated and 32 were completed in the control group. The treatment group underwentAn Mian Tietherapy half an hour before bed and the control group underwent placebo therapy in the same way of treatment group. Two groups were treated for 40 days and followed-up visit six months. The change of Pittsburgh Sleep Quality (PSQI), Index Insomnia Severity Index (ISI) and TCM Syndromes Scale were detected.Results The clinical total effective rate of treatment group was 72.7% and the control group was 9.4%, and the difference was statistically significant (χ2=46.977,P<0.01). The PSQI scores after treatment (7.55 ± 1.52vs. 13.90 ± 2.44,t=148.165), and follow up 1 month (8.97 ± 2.51vs. 13.17 ± 2.79,t=37.926) in the treatment group were significantly lower than those in the control group (P<0.01). The ISI scores after treatment (7.03 ± 3.37vs. 20.89 ± 4.40,t=73.75), and follow up 1 month (9.81 ± 3.16vs. 19.41 ± 3.66,t=40.79) in the observation group were significantly lower than those in the control group (P<0.01). The TCM Syndromes Scale scores after treatment (2.05 ± 1.09vs. 6.98 ± 1.23,t=17.116), and follow up 1 month (4.06 ± 1.59vs. 6.83 ± 0.91,t=68.055) and follow up 6 month (5.12 ± 1.84vs. 7.19 ± 1.07,t=27.716) in the observation group were significantly lower than those in the control group (P<0.01).Conclusions Acupoint application could obviously change the sleep quality and Chinese medicine symptom in chronic insomnia of Liver-stagnation and Spleen-deficiency Syndrome.

Objective To evaluate the value of low-density lipoprotein-cholesterol (LDL-C) and non-high-density lipoprotein-cholesterol (non-HDL-C) in differential diagnosis of familial hypertriglyceridemia (FHTG) and familial combined hyperlipidemia (FCHL).Methods We recruited 9 FHTG pedigrees (94 subjects) and 24 FCHL pedigrees (94 subjects) and then divided them into affected groups and non-affected groups according to lipid abnormality.Another 10 normal control pedigrees (57 subjects) served as controls.We compared the routine lipid levels such as triglyceride (TAG),total cholesterol (TC),HDL-C and LDL-C and non-HDL-C between the groups.After stratification based on TAG level,we observed the relationship between LDL-C and non-HDL-C.Last we confirmed and analyzed the cut-off value of differential diagnosis between FHTG and FCHL with receiver operating characteristic (ROC) curve.Results The levels of TAG,TC,and non-HDL-C were significantly higher in the affected group of FHTG than in the non-affected group of FHTG and the normal group (P＜0.01 or P＜0.05).The levels of TAG,TC,HDL-C,LDL-C and non-tHDL-C wcrc significantly higher in the affected group of FCHL than in the non-affected group of FCHL and the normal group (P＜0.01 or P＜0.05).The levels of TAG were significantly higher (P＜0.01) while TC,HDL-C,LDL-C and non-HDL-C levels were significantly lower (P＜ 0.01 or P＜0.05) in the affected group of FHTG than in the affected group of FCHL.The association between LDL-C and non-HDL-C was positive both in FHTG and FCHL,but the relationship became weaker as TAG level increased.The cut-off value of LDL-C and non-HDL-C was 3.575 mmol/L and 4.525 mmol/L,respectively.Conclusion In addition to the routinely used lipid indexes,non-HDL-C may be a new index for differential diagnosis of FHTG and FCHL,and may be superior to LDL-C in this regard.

Chemical constituents of Chlorella sorokiniana were isolated and purified by repeated column chromatographies, over silicagel and Sephadex LH-20. Their structures were identified on the basis of physicochemical properties and spectroscopic data analysis. Five compounds were obtained from the petroleum ether extract of Chlorella sorokiniana, and their structures were identified as (22E, 24R)-5alpha, 3beta-epidioxiergosta-6, 22-dien-3beta-ol(1),(24S)-ergosta-7-en-3beta-ol(2), loliolide(3), stigmasta-7,22-dien-3beta,5alpha,6alpha-triol(4), and 3beta-hydroxy-5alpha,6alpha-epoxy-7-megastigmen-9-one(5). The main liposoluble fractions from Chlorella sorokiniana maiuly contain fatty acids, alkyl acids and olefine acids. Components 1-5 were isolated from the genus Chlorella for the first time.
Biological Factors ; chemistry ; Chlorella ; chemistry ; Gas Chromatography-Mass Spectrometry ; Molecular Structure

Objective To explore the sero-prevalence and risk factors for herpes simplex virus 2 (HSV-2) infection and unprotected sexual behavior in an ethnically diverse population of HIVinfected subjects in a county of Yunnan province. Methods HIV-infected individuals attending for routine follow-up by local Center for Disease Control and Prevention (CDC) were recruited to participate in the study under 'informed consent'. A face-to-face questionnaire interview was administered to each participant. Blood was drawn for HSV-2 testing by HerpeSelect HSV-2 ELISA (Focus Diagnostics) and CD4+ T counting. Results A total of 300 HIV-infected individuals participated in the study. The mean age of the subjects was 37.6 years with 76.7% as males. Ethnically, Han, Dai and Jingpo accounted for 44.3%, 37.3% and 16.0% of the sample, respectively. Half of the subjects reported HIV acquisition through injection drug use. The sero-prevalence of HSV-2 was 35.0%. Results from multiple logistic regression analysis indicated that individuals who acquired HIV through heterosexual contact were more likely to be HSV-2 positive than those who acquired HIV through injection drug use (OR=4.244,95%CI: 1.924-9.364),whereas Dai (OR=0.300,95% CI: 0.152-0.593) and Jingpo (OR=0.376, 95% CI: 0.167-0.850) were less likely to be HSV-2 positive than the Hans. Among 105 people who were co-infected with HIV/HSV-2, 60 had sexual intercourses in the past 3 months and 41.7% of them reported no or inconsistent use of condoms. Most unprotected sexual contacts occurred within married couples. Conclusion HSV-2 infection was highly prevalent among HIV-infected individuals in this county, and a significant proportion of HIV/HSV-2 co-infected subjects engaged in unprotected sex. HSV-2 testing, behavioral and biomedical interventions among HIV-infected individuals and their sexual partners should be involved in the local HIV prevention and control programs.

The gene of mutated TNF-?D4 gene was amplified by overlap PCR and cloned into the prokaryotic expressive vector pBV220.TNF-?D4 contains two changes:substitutions of Pro8Arg,Ser9Lys,Asp10Arg,Ile157Phe,Leu29Ser,Arg31Val and a deletion of the N terminal four amino acids.The recombinant vector pBV220-TNF-?D4 was transformated into E.coli strain DH5?,and the high expression strain was obtained by screening monoclones.The level of expression was about 45% of total cell protein.After purification,the purity of fusion protein was above 90% by HPLC and relative ability was 8 ?107.TNF-?D4 was modificated by mPEG-ButyrALD。After purification,the purity of mPEG-TNF-?D4 was above 85% and relative ability was 8.6?107.The in vivo systemic toxicity of mPEG-TNF-?D4,which is indicated by LD50,is lower than that of rhTNF-?.These results strongly supported for the further study and exploitation of TNF-antitumor drug.

Objective To explore the clinical effect and toxicity of daunorubicin combined with cytarabine (DA regimen) and idarubicin combined with cytarabine (IA regimen) for the treatment of patients with acute myeloid leukemia (AML) as induction chemotherapy. Methods The clinical data of 84 newly diagnosed AML patients (except M3) treated with DA or IA regimen were analyzed retrospectively. DA regimen group included 32 patients (17 males and 15 females with median age of 46 years), while IA regimen group included 52 patients (29 males and 23 females with media age of 49 years). Efficacy index was complete remission (CR), total efficiency and adverse reactions after one course of chemotherapy rate. Results In DA regimen group,the CR rate was 65.6 %(21/32), and the total efficiency rate was 75.0 %(24/32), while in IA regimen group, the CR rate was 71.2 %(31/52), and the total efficiency rate was 80.8 %(42/52), respectively, but, the differences of media survival and 5-year survival rate were not statistically significant (16.8 months vs. 24.9 months, 26 % vs. 44 %, both P>0.05). The main side effect in the two groups included hematologic (bone marrow suppression) and non-hematologic adverse reactions, with no significant difference between the two groups (all P>0.05). Conclusion For newly diagnosed AML patients, remission rate and total efficiency of DA regimen are same as IA regimen after one course treatment, and adverse events between the two regimens do not differ significantly.

OBJECTIVE CYP2D is one of the most abundant subfamily of CYPs in the brain, especially in the cerebellum. Brain CYP2D is responsible for the metabolism of endogenous neurotransmitters such as tyramine and serotonin. Our previous studies have shown brain CYP2D can be regulated by exogenous and endogenous substances with tissue- specificity. The purpose of this study is to examine the effects of cerebral CYP2D on the mice behavior and the regulatory mechanism of brain CYP2D by growth hormone. METHODS Mice received the stereotaxic injection with CYP2D inhibitor quinine in deep cerebellar nuclei of cerebellum. The animals were tested with rotarod apparatus, balance beam, water maze, elevated plus maze and open field. The changes in CYP2D22, PPARαand PPARγ in brain regions and liver were assayed in male growth hormone receptor knockout mice, SH-SY5Y cells and HepG2 cells. RESULTS The inhibition of cerebellum CYP2D significantly affected the spatial learning and exploring ability of mice. Compared with WT mice, CYP2D expression was lower in brain regions from GHR(-/- ) male mice; however, hepatic CYP2D level was similar. Pulsatile GH decreased PPARα mRNA level, and increased mRNA levels of CYP2D6 and PPARα in SH- SY5Y cells. In HepG2 cells, pulsatile GH resulted in decreases in PPARα and PPARγ mRNA levels, but not CYP2D6. PPARα inhibitor induced CYP2D6 mRNA and protein by 1.32-fold and 1.43-fold in SH-SY5Y cells. PPARγ inhibitor decreased CYP2D6 mRNA and protein by 74.76% and 40.93%. PPARα agonist decreased the level of CYP2D22 mRNA in liver and cerebellum, while PPARγ agonist rosiglitazone resulted in diametrically increases. The luciferase assay showed that PPARγ actived the CYP2D6 gene promoter while PPARα inhibited its function. Pulsatile GH declined the binding of PPARα with CYP2D6 promoter by 40%, promoted the binding of PPARγ with CYP2D6 promoter by approximate 60%. The levels of brain and liver PPARα expression in male GHR(-/- ) mice is obviously higher than those in WT mice. The level of PPARγ in male GHR(-/- ) mice was decreased in the frontal cortex and hippocampus, while remained stable in the cerebellum and striatum; meanwhile, PPARγ was increased in the liver. CONCLUSION Brain CYP2D may be involved in learning and memory functions of central system. Masculine GH secretion altered the PPARs expression and the binding of PPARs to CYP2D promoter, leading to the elevated brain CYP2D in a tissue- specific manner. Growth hormone may specifically alter the metabolic and synthetic of important endogenous substances in the central nervous system (such as serotonin) through the specific regulation of brain CYP2D expression.