In the present study, a new compound named 17-(6-cinnamamido-hexylamino-)-17-demethoxygeldanamycin (CDG) was obtained by introducing the cinnamic acid (CA) group into the 17-site of geldanamycin (GDM). The anti-cancer effects of CDG in vitro and in vivo were evaluated. MTT assay was used to examine the inhibitory effect of CDG on the proliferation of MCF-7, HepG2, H460 and SW1990 cells. Immunofluorescent staining flow cytometry combined with Annexin V-FITC/PI staining were used to detect apoptotic cells. Transwell assay was used to analyze the effect of CDG on cell invasion and migration ability. Western blotting was used to detect the expression levels of RAF-1, EGFR, AKT, CDK4 and HER-2 of MCF-7, HepG2 and H460 cells. The toxicities of CDG and GDM were evaluated in mice. Using the subcutaneously transplanted MCF-7 xenograft in nude mice, inhibitory effect was evaluated in vivo. The results showed that CDG inhibited the proliferation of cancer cells (IC50: 13.6-67.4 microg.mL-1). After exposure to CDG for 48 h, most cells presented typical morphologic changes of apoptosis such as chromatin condensation or shrunken nucleus. The rates of apoptosis of MCF-7, HepG2, H460 and SW1990 cells incubated with 10 microg.mL-1 CDG were 23.16%, 27.55%, 22.21%, 20.47%, respectively. A dose-dependent reduction of migration of four cell lines was found after exposure to CDG. The decreased levels of RAF-1, EGFR, AKT, CDK4 and HER-2 showed that CDG possessed HSP90 inhibitory effect. The result of animal toxicity test on the mice suggested that CDG had lower toxicity than GDM. Meanwhile, CDG inhibited the growth of MCF-7 xenografts of athymic mice.
Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Benzoquinones ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase 4 ; metabolism ; Female ; HSP90 Heat-Shock Proteins ; antagonists & inhibitors ; Humans ; Lactams, Macrocyclic ; chemical synthesis ; chemistry ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Proto-Oncogene Proteins A-raf ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Random Allocation ; Receptor, Epidermal Growth Factor ; metabolism ; Receptor, ErbB-2 ; metabolism ; Tumor Burden ; drug effects ; Xenograft Model Antitumor Assays

24 strains obtained from root nodules of Psoralea corylifolia, Pueraria lobata, and Campylotropis macrocarpa of Yunnan province were studied with numerical taxonomy and 16S rDNA PCR-RFLP. Results of numerical taxonomy indicated that all strains included 10 reference strains were divided into 3 groups at 84% similarity. Group III is an unknown group with no reference strains. Group I is slow-growing kind, and group II fast and middle-slow-grower. The dendrogram derived from 16S rDNA PCR-RFLP showed that all strains divided into five phylogenetic branches at the similarity of 70%. They are branches I and V with no reference strains, Agrobacterium-Sinorhizobium-Rhizobium, Mesorhizobium and Bradyrhizobium. Not all results of numerical taxonomy are accord with 16S rDNA PCR-RFLP, and 2 strains at the same group with A. tumefaciens IAM13129T.

OBJECTIVETo establish HPLC fingerprint of HQSM capsules,and a new method for correlativity analysis between fingerprint peaks of HQSM and its relevant herbs based on the fingerprint analysis.

METHODThe chromatographic fingerprints of HQSM and its relevant herbs were analyzed by HPLC configured with DAD. The similarity of the HPLC fingerprints between HQSM and its relevant herbs and the comparison of UV spectra of the main peaks in both of fingerprints were used as indices to evaluate the correlativity.

RESULTThe HPLC fingerprint with 16 common peaks was established and the chromatographic peaks were determined.

CONCLUSIONThe established method effectively controlled the quality of HQSM and establised the correlativity between HQSM and its relevant herbs, which would contribute to deduce the chemical components or sources of traditional Chinese medicine prescription.

Capsules ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the changes of human leukocyte antigen DR (HLA-DR) expression in the monocytes of elderly patients with sepsis.

METHODSA total of 213 elderly patients were divided into 4 groups according to the criteria defined by the American College of Chest Physicians/ Society of Critical Care Medicine (ACCP/SCCM), namely the non-sepsis group (group A), sepsis group (group B), severe sepsis group (group C) and septic shock group (group D). Another 60 healthy subjects were recruited as the control group. HLA-DR expression in the monocytes from each subject was determined by flow cytometry, and the HLA-DR levels were compared between the groups.

RESULTSCompared with that of the healthy control, HLA-DR expression was significantly increased in group A and lowered in group D. No significant differences were found between the control group and groups B and C. HLA-DR expression was significantly different between groups A, B, C, and D (P<0.01), increasing in the order of group D (32.74-/+13.4)%, group C (61.9-/+14.29)%, group B (69.99-/+12.8)%, and group A (85.06-/+15.37)%.

CONCLUSIONSAlthough with multiple organ diseases and repeated or long-term hospitalization, the non-septic elderly patients do not show lowered expression of HLA-DR. In the elderly patients with sepsis and severe sepsis, HLA-DR expression is not a sensitive index, but obviously lowered HLA-DR expression may serve as a specific indicator in elderly patients with septic shock.

Aged ; Aged, 80 and over ; Biomarkers ; analysis ; Female ; Flow Cytometry ; HLA-DR Antigens ; analysis ; biosynthesis ; Humans ; Leukocytes, Mononuclear ; immunology ; Male ; Sepsis ; immunology ; Shock, Septic ; immunology

Adolescent ; Child ; Cord Blood Stem Cell Transplantation ; Female ; Flow Cytometry ; Humans ; Leukocyte Count ; Male ; Siblings ; Transplantation, Homologous ; immunology ; Treatment Outcome

To explore the Fas and mdr-1 expression and their correlation in multidrug resistance (MDR), Fas and mdr-1 expressions of bone marrow from 59 patients with newly diagnosed AL before therapy and after complete remission were detected by direct immunofluorescence with flow cytometry and semi-quantitative RT-PCR, respectively. The results showed that in newly diagnosed AL patients, Fas expression in AML was higher than in ALL (P < 0.05), mdr-1 expression in AML and ALL had no difference (P > 0.05), the expressions of Fas and mdr-1 had correlation (r = -0.282, P < 0.05) in AL; the results of simple-variable and multivariable COX survival factor model analysis suggested that Fas and mdr-1 expressions were prognostic factors for the effect of therapy and survival. Log rank test, comparing the groups of Fas(+) with Fas(-), mdr-1(+) with mdr-1(-), demonstrated that the CR rates and median remission time of every two groups had significant difference. It is concluded that in AL, Fas and mdr-1 expressions have high correlation with the effect of treatment, Fas expression probably is one of the favorable prognostic factors, mdr-1 is an unfavorable prognostic and less effective factor.
Adolescent ; Adult ; Aged ; Child ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Genes, MDR ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; metabolism ; mortality ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; metabolism ; mortality ; RNA, Messenger ; analysis ; fas Receptor ; analysis

OBJECTIVETo explore the clinical classification method of keloids and providing a thread for the treatment of keloids.

METHODSTo summarize the 600 cases of keloid patients we accepted and diagnosed from November 2004 to October 2012, and filling in keloid patients information sheet, recording the keloids form by photographs, analyzing the treatment, putting forward the classification method of keloids in clinic.

RESULTSAccording to the position and quantity that keloids grow, the keloid patients are divided into four major categories:one in single site, one in each site, more than one in single site and more than one in each site; According to the area and thickness of keloids, the keloid single lesion is divided into four subclasses: type of small area and thin, type of small area and thick, type of large areas and thin,type of large areas and thick; According to the number of lesions, keloid multiple lesions is divided into two subgenera: isolated multiple and dispersion multiple, different kinds of keloids suit different methods of treatment.

CONCLUSIONThe clinical classification method of keloids can be used to provide thought for the treatment of keloids, and have a good application value.

Humans ; Keloid ; classification ; pathology ; therapy

In order to increase the plasma half-life and tissue specificity of IL-1 receptor antagonist, a recombinant fusion protein IL-1Ra-HSA, linked by a rigid peptide linker PAPAP, was engineered and expressed by the Pichia pastoris host cells. The fusion protein was secreted to the host cells culture, identified by Western blot, and purified by affinity chromatography. This was followed by a further examination of its bioactivity and pharmacokinetics. Our results demonstrated that the fusion protein retained the antagonist activity of IL-1Ra, capable of binding specifically to the IL-1 receptor on human melanoma A375.S2 cells, and inhibits the cytolytic effect of IL-1beta to A375.S2 cells. Albumin fusion dramatically extended the half-life of IL-1Ra and resulted in a specific accumulation of IL-1Ra in the arthritic paws and a lower distribution of IL-1Ra in other organs such as liver, kidney, spleen and lung in mice with collagen-induced arthritis. The findings reported herein indicate that the fusion protein is likely to have greater clinical applications in areas such as the treatment of rheumatoid arthritis.
Animals ; Apoptosis ; drug effects ; Arthritis, Experimental ; metabolism ; Cell Line, Tumor ; Forelimb ; metabolism ; Half-Life ; Humans ; Interleukin 1 Receptor Antagonist Protein ; genetics ; metabolism ; pharmacokinetics ; pharmacology ; Interleukin-1beta ; toxicity ; Male ; Melanoma ; pathology ; Mice ; Mice, Inbred DBA ; Pichia ; genetics ; metabolism ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacokinetics ; pharmacology ; Serum Albumin ; genetics ; metabolism ; pharmacokinetics ; pharmacology ; Tissue Distribution

Objective MR microscopy technique was used to study the visualization of senile plaque deposition in brains of the Alzheimer disease(AD)transgenic mice.Methods Two transgenic mice and 2 wild type mice at the age of 17 months were scanned in vivo using T_2 weighted image.After MR imaging,the brains were cut serially and immunostained according to the orthogonal pilot images.MR T_2 weighted images and immunohistological images of the senile plaque were observed and matched.Results The MR images showed that some black spots were visible in the hippocampus and cerebral cortex of the AD transgenic mice and some spots were consistent with the senile plaques on immunohistological sections.There were no spots in the MR images and the immunohistological sections of the wild type mice.Conclusion It is possible that MR microscopy can be used to detect the deposition of the senile plaque and diagnose AD specifically.

Objective To detect the presence of ion channel protein and its role in cell growth and proliferation in human head and neck squamous carcinoma ceils(SCC).Methads Human head and neck squamous carcinoma SCC-25 cell line was tasted with transient receptor potential melastatin 7(TRPM7) antibody using the method of immunocytochemistry.The role of TRPM7 in cell growth and proliferation was evaluated through its blockade by ion channel blockers and specific siRNA using lactate dehydrogenase (LDH)assay technique.Results Clear immunoreactivity against TRPM7 was detected in almost all SCC-25 cells tested,whereas no immunoreactivity was observed in negative contr01.The inhibitory effect of Gd3+,a non-specific ion channel blocker,on cell growth and proliferation was potenL Addition of growth of SCC-25 cells,as compared with control cells growing in normal medium(t was 4.1414 and 6.2661.P was 0.0256 and 0.0082 respectively).However,the effect of 2-APB was striking.Cell n=16)compared with cells growing in normal medium.Suppression of TRPM7 expression by siRNA also significantly inhibited the growth and proliferation of these cells(t=4.3446,P=0.0002,n=32,compared) with nontransfected ceHs).whereas cells transfected with negative control siRNA showed no difierence in cell proliferation compared with nontransfected cells.Conclusions All of those results strongly suggest the existence of TRPM7 channel in human head and neck squamous carcinoma cells.Ion channel blockers serve as a potent inhibitor of SCC-25 cell proliferation.The striking inhibitory effect of 2-APB on cell growth and proliferation may promise clinical workers an inspiring remedy for fighting against carcinoma.