HSP90, which is the biomarker of cell stress and endogenous protective protein, functions as a molecular chaperone. Many client proteins of HSP90, including EGFR, Met, Raf-1, IKK and p53, play important roles in the occurrence and development of tumor. Binding of HSP90 inhibitors triggers the deactivation of HSP90, resulting in client protein degradation, and hence inhibits the tumor growth by blocking multiple targets involved in signaling of tumor proliferation. This review summarizes recent development of small molecule inhibitors bound to N-terminal of HSP90.
Antineoplastic Agents ; chemistry ; HSP90 Heat-Shock Proteins ; antagonists & inhibitors ; Humans ; Neoplasms ; Signal Transduction

Aged ; Ear Neoplasms ; pathology ; Ear, Middle ; pathology ; Female ; Hemangioendothelioma, Epithelioid ; pathology ; Humans

α-Hederagenin (H), derived from Hedera nepalensis var.sinensis, is a pentacyclic oleane-type triterpenoid that exhibits clear cytotoxicity to different tumor cell lines.In this study,a series of novel C-28 derivatives of hederagenin (H) were designed and synthesized in attempt to develop potent tumor resistance reverse activities agents. Previous research showed that H6 displayed robust reverse activity for paclitaxel resistance in KBV cells. Importantly, Co-treatment of paclitaxel with H6 significantly reduced the tumor weight to 42%. Pleasingly, H6 enhanced the efficacy of paclitaxel against KBV cancer cell-derived xenograft tumors in nude mice.Mechanism studies had found that H6 activated permeability glycoprotein(P-gp)ATPase,reduced intracellular ATP levels and inhibited efflux of P-gp substrates,thus enhancing the antitumor activity of paclitaxel on KBV cells.Molecular docking analysis of homology P-gp and H6 then conducted using the Surflex-Dock module.H6 showed a high binding affinity docking score with a total score of 5.4148,much higher than that of H(0.1414).The nov-el C-28 derivatives of H was synthesized from H6 via three-step reaction. The reversal activity of all synthesized H derivatives were tested using the MTT assay.The results showed that the derivatives of nitrogen groups at C-28 displayed same even potent activity than parent compound H6.In addition,its underlying mechanism of action and in vivo activity are in explore.

Based on the gene sequence of AIV matrix protein 2(M2) and two cytotoxic T-lymphocyte epitopes derived from nucleoprotein,the prokaryotic expression vector pET-3M2e-NP1-NP2 was constructed.The target gene was expressed in the solvable form in BL21(DE3) when induced with 1.0 mmol/L IPTG and Western blot analysis showed that the expression product had good immunogencity.Purified fusion protein was mixed with various amount of adjuvant,including Freund's,Vash oil and chitosan,and then immunized 20-day-old chicken by intramuscular injection and boosted 3weeks later.Blood samples were collected weekly following the primary vaccination.The anti-M2e antibody was detected with ELISA method with the synthesized peptide as coated antigen;the neutralizing ability of anti-serum was evaluated on MDCK cell line and chick embryo,the CD4+ and CD8+ T lymphocyte amounts in peripheral blood of immunized chicken was measured by flow cytometry.Results showed that the fusion protein can induce immunological reaction,and the antibody can bind with the viral M2 protein that expressed on the surface of MDCK cells.Flow cytometry result showed that CD4+ and CD8+ T lymphocyte in peripheral blood increased obviously following immunization(P

0.05).Conclusions The higher level of SEB-specific IgM and IgE in AD and eczema indi cates the colonization of Staphylococcus aureus,which participates in the exace rbation of allergic inflammation,is involved in the pathogenesis of AD and ecz ema.

It is a challenging and important project to prolong the in vivo half life of protein and peptide drugs by physicochemical methods without new molecular entities generation. Protein crystallization provides a new strategy for improving the stability and in vivo delivery of these drugs. We show here that recombinant human interferon-alpha (rhIFN) can form spherical crystals. The physical and chemical features of the crystals were characterized, and drug dissolution was determined in vitro. The pharmacokinetics of crystallized interferon after sc injection in rabbit at 1.5 x 10(7) U x kg(-1) was compared to that of soluble form. The crystals were characterized as mono-dispersed spheres, with yield of > 80%, mean diameter size of about 16 microm and crystallinity of 23.2%. The in vitro dissolution behavior of crystallized rhIFN was featured as low initial burst release (21% within the first 2 h) and prolonged cumulative dissolution time up to 72 h without biological potency lost. After sc administration of soluble and crystallized interferon in rabbits, the peak time (T(max)) and half life (t1/2) were prolonged from (1.80 +/- 0.45) h and (1.35 +/- 0.35) h to (13.20 +/- 2.68) h and (10.68 +/- 1.97) h, respectively. The corresponding peak concentration decreased from (1 411.10 +/- 575.28) U x mL(-1) to (721.37 +/- 206.55) U x mL(-1). PK/PD analysis indicated that (96.87 +/- 20.30) % of relative bioavailability was obtained. The research results of this work will provide important academic value and application prospect for improving clinical therapeutic effect and development of biomacromolecules delivery system for protein and peptide drugs.
Animals ; Antiviral Agents ; administration & dosage ; chemistry ; pharmacokinetics ; Biological Availability ; Crystallization ; Delayed-Action Preparations ; Drug Delivery Systems ; Half-Life ; Humans ; Injections, Subcutaneous ; Interferon-alpha ; administration & dosage ; chemistry ; pharmacokinetics ; Male ; Rabbits ; Recombinant Proteins ; administration & dosage ; chemistry ; pharmacokinetics ; Solubility ; Surface Properties

AIMTo determine the protective effect of recombinant human interleukin-2 (rhIL-2) in inhalant form on experimental respiratory tract infection with Klebsiella pneumoniae in mice.

METHODSMice were infected with the method of nasal intubation drip. During infection, mice were given rhIL-2 by sc injection and the method of nasal intubation drip. There were normal group, vehicle group, model group, rhIL-2 groups and gentamicin group. In the end, the pathological changes in the lung were observed. The survival time and the mortality within a week of each group were recorded. The total protein content, the albumin content, the activity of alkaline phosphatase and the activity of lactic dehydrogenase of broncho-alveolar lavage fluid (BALF) were dertermined and compared.

RESULTSSymptoms of Klebsiella pneumoniae were remarkably relieved because of rhIL-2 administration. The total protein content, the albumin content, the activity of alkaline phosphatase and the activity of lactic dehydrogenase of BALF were less than those in the vehicle group and the model group.

CONCLUSIONInhalation of rhIL-2 can alleviate the pathological changes in the lung after infection. At the same dose, it could be seen that the effect of rhIL-2 in inhalant form was better than that of the injection.

Administration, Inhalation ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Female ; Interleukin-2 ; administration & dosage ; pharmacology ; Klebsiella Infections ; drug therapy ; metabolism ; pathology ; Klebsiella pneumoniae ; Lung ; pathology ; Mice ; Mice, Inbred ICR ; Recombinant Proteins ; administration & dosage ; pharmacology

OBJECTIVETo investigate the effect and mechanism of Compound Salvia injection (CSI) on nitrate ester tolerance.

METHODSEighty-four patients with coronary heart disease (CHD) were randomly divided into three groups, Group A treated with isosorbide dinitrate (ISD, 15 mg, 4 times per day) alone, Group B with ISD plus CSI and Group C with ISD plus vitamin C. The therapeutic course for all groups was 10 days. The tolerance to nitrate ester and blood pressure were monitored. Before and after treatment, the color Doppler ultrasonic apparatus was used to detect the baseline value of humeral arterial internal diameters (D0), the humeral arterial dilatory response under compression [D1, that is, the flow-mediated vasodilation (FMD)] and the vasodilatory response after sucking of nitroglycerin (D2). And the blood levels of endothelin-1 (ET-1), endothelial nitric oxide synthase (eNOS) mRNA expression were determined. The endothelial-dependent vasodilation (EDD) was expressed by (D1 - D0)/D0 x 100%, and the endothelial-independent vasodilation (EID) was expressed by (D2 - D0)/D0 x 100%.

RESULTS(1) The occurrence rate of nitrate tolerance in Group B and C (28.57% and 35.7%) was lower than that in Group A (64.29%), but insignificant difference was found between the former two. (2) After treatment, blood pressure increased in Group A to the level of pre-treatment, that in Group C also increased but still lower than that of pre-treatment, while insignificant increase was observed in Group B, comparison between Group B and C showed significant difference (P < 0.05). (3) After treatment, EID lowered in Group A, EDD increased in Group B and C (P < 0.05), EDD and EID in Group B and C were higher than those in Group A (P < 0.05), and EDD was higher in Group B than in Group C (P < 0.05). (4) After treatment, ET-1 mRNA expression lowered in Group B, eNOS mRNA expression increased in Group B and C, with significant difference as compared with those before treatment and those in Group A (P < 0.05), and eNOS mRNA expression in Group C was lower than that in Group B (P < 0.05).

CONCLUSIONCSI could partially prevent the occurrence of tolerance to nitrate ester, with the effect better than vitamin C, the mechanism might be related with its regulation on eNOS, ET-1 mRNA expression and protection on vascular endothelial function.

Adult ; Aged ; Coronary Disease ; drug therapy ; Drug Resistance ; Drugs, Chinese Herbal ; administration & dosage ; Endothelin-1 ; biosynthesis ; genetics ; Female ; Humans ; Injections, Intravenous ; Isosorbide Dinitrate ; therapeutic use ; Male ; Middle Aged ; Nitric Oxide Synthase ; biosynthesis ; genetics ; Nitric Oxide Synthase Type III ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Salvia miltiorrhiza ; Vasodilator Agents ; therapeutic use

OBJECTIVETo investigate the changes of drug sensitivity of spindle poison-induced polyploid tumor cells to chemotherapeutic agents and its possible mechanism.

METHODSNocodazole in a dose of 100 ng/ml was used to induce polyploidization in a breast cancer cell line MDA-MB-231 cells. The polyploid cells (T-MDA-MB-231) were sorted by flow cytometry. The morphological changes and proliferation of T-MDA-MB-231 cells were compared with that of MDA-MB-231 cells. The cell growth inhibition was assessed by MTT assay. The cells were treated with paclitaxel, docetaxel, vincristine, epirubicin, 5-Fu, VP16 and oxaliplatin, respectively. Those cells were labeled with annexin V-FITC/PI and analyzed by flow cytometry. Bcl-2 was knocked down in T-MDA-MB-231 cells using SiRNA and their growth inhibition was evaluated by MTT assay to evaluate the reversing effect of Bcl-2-silencing on drug resistance.

RESULTSThe polyploid T-MDA-MB-231 cells grew in vitro continuously and maintained constant DNA content. They had a larger cell size, and grew more slowly than MDA-MB-231 cells. The IC(50(s)) of T-MDA-MB-231 cells were significantly higher than that of the MDA-MB-231 cells: paclitaxel: (6.37 ± 0.07) vs. (2.05 ± 0.83) µmol/L; docetaxel: (32.98 ± 1.48) vs. (11.95 ± 0.98) µmol/L; vincristine: (35.28 ± 1.66) vs. (14.58 ± 0.94) µmol/L; oxaliplatin: (19.07 ± 0.45) vs. (9.75 ± 1.05) µmol/L; 5-Fu: (85.49 ± 3.21) vs. (31.35 ± 1.51) µmol/L; and epirubicin: (0.53 ± 0.06) vs. (0.15 ± 0.01) µmol/L, (all P < 0.05). The IC(50(s)) of VP16 in T-MDA-MB-231 cells was (2.85 ± 0.50)µmol/L, significantly lower than the (12.20 ± 1.55) µmol/L in MDA-MB-231 cells (P < 0.05), and that of T-MDA-MB-231 cells after Bcl-2-knocked down by siRNA was (19.59 ± 0.48) µmol/L, significantly higher than the (12.20 ± 1.55) µmol/L in the MDA-MB-231 cells (P < 0.05). The IC(50(s)) of docetaxel of T-MDA-MB-231 cells after Bcl-2-knocked down by siRNA was (21.52 ± 0.68) µmol/L, significantly decreased and lower than that before Bcl-2 silencing (32.98 ± 1.48) µmol/L.

CONCLUSIONSOur results indicate that polyploid tumor cells induced by spindle poison Nocodazole are more resistant to most of chemotherapeutic drugs. Downregulation of Bcl-2 increases the sensitivity of polyploid cells to docetaxel. The high expression of Bcl-2 may be one of the drug resistance mechanisms of polyploid tumor cells. The polyploid tumor cells are relatively sensitive to VP16, suggesting that VP16 might be an effective candidate drug for treatment of chemoresistant polyploid tumors.

Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Drug Resistance, Neoplasm ; Epirubicin ; pharmacology ; Etoposide ; pharmacology ; Female ; Fluorouracil ; pharmacology ; Gene Knockdown Techniques ; Humans ; Inhibitory Concentration 50 ; Nocodazole ; pharmacology ; Organoplatinum Compounds ; pharmacology ; Paclitaxel ; pharmacology ; Polyploidy ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Taxoids ; pharmacology ; Vincristine ; pharmacology

OBJECTIVETo estimate the number of drug users in Hongjiang, Hunan and to develop strategy for drug reduction in the future.

METHODSTwo capture-recapture methods were used to estimate the numbers of drug addicts. Random stratified sampling survey was used to verify the optimum allocation. The first capture-recapture method (CR1) referred to the number from optimum allocation random stratified sampling survey conducted in the communities and the number from local Public Security Bureau list being the second capture. The second capture-recapture method (CR2) referred to the collection of records in the detoxification unit with an interval of 4 months. The estimated number was calculated under Seber's adjustment formula. Face to face interview was carried out during the optimum allocation random stratified sampling survey process.

RESULTSOf 1388 interviewed in the communities, 24 (1.73%) were identified as drug addicts under the optimum allocation random stratified sampling survey. When the figure 1.73% was applied to the total population (72,709) in Hongjiang, the result yielded an estimation of 1258 drug addicts. The estimated numbers of CR1 and CR2 were 904 and 1069 respectively. However, the number was 1.3 to 1.6 fold higher than the reported number (687) by local Public Security Bureau.

CONCLUSIONThe capture-recapture method seemed a better method in estimating the number of drug addicts.

China ; epidemiology ; Female ; Humans ; Incidence ; Male ; Needle Sharing ; Population Surveillance ; methods ; Substance Abuse, Intravenous ; epidemiology