Objective: To study the chemical constituents of Humulus scandens. Methods: The compounds were isolated and repeatedly purified by silica gel, TLC, and Sephadex LH-20 column chromatography, and their structures were elucidated on the basis of physicochemical constants and spectral analysis. Results: Eleven compounds were obtained and elucidated as (24R)-stigmast-7, 22(E)-dien-3β- ol (1), daucesterol (2), stigmast-3, 6-dione (3), epidioxyergosta-6, 22-dine-3β-ol (4), stigmasterol (5), soya-erebroside II (6), oleanolic acid (7), bis-(2-ethylhexyl) phthalate (8), scopoletin (9), betulimicaciol (10), and soya-erebroside I (11). Conclusion: The compounds 1, 3, 4 and 6-11 are isolated from the plants of Humulus Linn. for the first time.

Objective To use bioinformatics methods to analyze large amounts of data generated by gene chips and to screen common key genes in hepatocellular carcinoma in human and rat.Methods For search of the medical literature,3 sets of gene chip with data which met our predetermined criteria were downloaded from the GEO database.The data were standardized by using the bioconductor and R version of the 2.10.1 version.The original data of the affymetrix platform were normalized with background correction,standardized and transformed into log2 by using the algorithm of the affy packages RMA.The TTEST function of the excel was then used to calculate the significance of each gene.The DAVID was used for gene ID conversion and a table was established for samples and the corresponding gene expression data.A meta analysis was performed to calculate the common genes of human and rat.An enrichment regulation pathway was gained with the KEGG in the DAVID library. Results There were 26 common expression genes in the development process of hepatocellular carcinoma in human and rat.Five of these genes were up-regulation genes,while twenty-one were down-regulation genes.An enrichment pathway,which is a focal adhesion pathway,was found and this pathway has been reported to be associated with development of hepatocellular carcinoma.Conclusion With bioinformatics,we were able to screen common key genes and a pathway which were closely related to development of hepatocellular carcinoma in human and rat.

Objective To elucidate the effects of SP600125 at different concentrations on the proliferation and osteo-differentiation of human adipose-derived stem cells (hASCs).Methods The hASCs harvested were cocuhured with SP600125 at concentrations of 0 μmol/L,1 μmol/L,5 μmol/L and 10 μmol/L in growth medium (OM group) and in osteogenesis medium (OM group),respectively.The DNA quantitative assay was carried out to evaluate proliferation of the hASCs;flow cytometry was used to determine the effect of SP600125 on the cell cycles of hASCs;Alkaline phosphatase level (ALP) and calcium deposition tests were conducted to observe the effects of SP600125 at different concentrations on osteogenic differentiation of the hASCs.Results The proliferation of hASCs was inhibited by 42.1％ when the cells were cocultured with SP600125 at the concentration of 10 μmol/L;the suppression decreased with decreased concentration of SP600125.The hASCs of phase G0/G1 in GM cocultured with SP600125 at the concentration of 10 μmol/L were more than those in GM cocultured with dimethylsulfoxide at the same concentration.ALP test revealed that after 10 days of culture in vitro the staining was more and more weakened and scattered and the ALP activity was more and more decreased with the increased concentration of SP600125.The extracellular calcium deposition of hASCs after 14 days of culture in vitro showed that the size and number of calcium nodules decreased with the increased concentration of SP600125.Conclusion SP600125 can suppress the proliferation and osteogenic differentiation of hASCs in vitro.

OBJECTIVETo evaluate the possibility that using intracoronary delivery of autologus bone marrow-derived mesenchymal stem cells (MSCs) to improve the cardiac function after acute myocardial infarction (AMI) in miniature pig.

METHODSMSCs were cultured in Dulbecco's modified Eagle's medium-F12 (DMEM/F12) medium. AMI model was made by blocking the blood stream of the first diagonal branch in miniature pig, and released the branch after 90 minutes. After 10-14 days, (4-6) x 10(7) culture-expanded autologus 4', 6-diamidino-2-phenylindole (DAPI)-labelled MSCs were transplanted into each host heart's AMI area through intracoronary way. Ultrasonic cardiography (UCG) was performed to observe the left ventricular function at 3 months after transplantation. The cellular transplanted hearts were harvested and investigated by immunohistochemical analysis.

RESULTSLeft ventricular function of the MSCs group was improved significantly 3 months later compared with the control group [(54.65 +/- 3.39) vs (43.98 +/- 4.21)%, (P < 0.01)]. Exogenous MSCs survived and site-differentiated into cardiomyocytes in infracted hearts.

CONCLUSIONMSCs can play a benificial role to repair damaged heart. Heart function can be improved after MSCs transplantation in porcine myocardial infarction model.

Animals ; Female ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Myocardial Infarction ; pathology ; physiopathology ; therapy ; Swine ; Swine, Miniature ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo explore the molecular mechanism of dutasteride inhibiting fertility by studying its effects on the expressions of the epididymal epithelial junction proteins Claudin1 and β-catenin in rats.

METHODSSixteen 3-month-old SD male rats were equally divided into an experimental and a negative control group to be treated intragastrically with dutasteride at 40 mg/kg per day and the same dose of solvent, respectively, for 14 consecutive days. Then, the sperm motility and morphology of the rats were detected by computer-assisted sperm analysis, the serum levels of testosterone (T) and dihydrotestosterone (DHT) measured by ELISA, changes in the tight junction of epididymal cells observed under the transmission electron microscope, the protein and gene expressions of Claudin1 and β-catenin determined by RT-PCR and immunohistochemistry, and the conception rate of the mated female rats calculated.

RESULTSDutasteride significantly suppressed the serum DHT level, sperm motility, and fertility of the rats (P <0.05). Interspaces between epididymal epithelial cell tight junctions were observed, the volume of epididymal fluid obviously increased, and the expressions of Claudin1 and β-catenin gene and protein remarkably downregulated in the experimental rats (P <0.05).

CONCLUSIONDutasteride can significantly inhibit the fertility of male rats by reducing the serum DHT level, suppressing Claudin1 and β-catenin expressions, and damaging epididymal epithelial cell junctions.

Animals ; Azasteroids ; pharmacology ; Claudin-1 ; metabolism ; Dihydrotestosterone ; blood ; Dutasteride ; Epididymis ; drug effects ; metabolism ; Female ; Fertility ; drug effects ; Humans ; Intercellular Junctions ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; drug effects ; Testosterone ; blood ; Urological Agents ; pharmacology ; beta Catenin ; metabolism

This study was purposed to analyse the immunophenotypic characteristics of chronic myelomonocytic leukemia (CMML), myelodysplastic syndromes (MDS) and acute monocytic leukemia (AML-M5b) by using multiparameter flow cytometry, and to explore its significance in diagnosis and differential diagnosis. The immunophenotypic characteristics of bone marrow samples from 14 CMML patients, 48 MDS patients, 46 AML-M5b patients and 18 normal persons were analyzed and compared by multiparametric flow cytometry. The results showed that the ratio of monocytes in CMML patients was obviously higher than that in MDS, AML-M5b patients and normal persons (P < 0.05), but there was no statistically significant difference between bone marrow samples of MDS and AML-M5b patients as well as normal persons. The ratio of blast cells in MDS patients was obviously higher than that in normal persons (P < 0.05), but did not show significant difference as compared with CMML patients. The ratio of mature granulocytes in AML-M5b patients was obviously lower than that in CMML and MDS patients as well as normal person bone marrow (P < 0.05). Certain differences of CD45/SSC characteristics in MDS, AML-M5b and CMML patients were found in comparison with normal persons. The abnormal expression of CD2, CD56, and CD14 tailing phenomenon were observed in CMML patients in comparison with bone marrow samples of MDS, AML-M5b and normal persons (P < 0.05). Lack and decrease of CD15 expression in MDS and CMML patients was significant different from AML-M5b and normal persons marrow, abnormal expression rate of CD15 in CMML patients was higher than that in MDS patients (P < 0.05), the CD13/CD11b/CD16 abnormal expression of granulocytes was seen in both CMML and MDS patients, but there was no statistically significant difference between them. Other antigens showed abnormality of varying degrees, but did not have any statistical significance. It is concluded that MDS, CMML and AML-M5b displayed a certain degree of similarity, and also possess their own immunophenotype characteristics. Comprehensive analysis of immunophenotype by multiparameter flow cytometry may be important for differential diagnosis among CMML, MDS and AML-M5b. High percentage of monocytes, abnormal coexpression of CD2, CD56 and CD14 tailing phenomenon, lack or decrease of CD15 as well as abnormal expression of CD13/CD11b/CD16 in granulocytes may play important roles in diagnosis of CMML.
Case-Control Studies ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; methods ; Leukemia, Monocytic, Acute ; diagnosis ; immunology ; Leukemia, Myelomonocytic, Chronic ; diagnosis ; immunology ; Myelodysplastic Syndromes ; diagnosis ; immunology

Translocating protein and translocating peptides have therapeutic potential against tumors by exposing the cytotoxic domains of toxic proteins to the cell cytosol. The aim of this study is to investigate the effect of N-terminally fused PE translocating peptides on granzyme B (GrBa) activity. PE II-GrBa fusion protein genes were constructed by replacing N-terminal signal and acidic dipeptide sequence of human granzyme B gene with two truncated translocating sequences of Pseudomonas exotoxin A (PE II aa 280-364/358) by recombinant PCR, and then cloned into pIND inducible expression vector. The resulting pIND-PE II-GrBa expression vectors were co-transfected with assistant plasmid pVgRXR into HeLa cells through lipofectamine, followed by selection on G418 and zeocin. The resistant cells were collected and induced with ponasterone A. Western blot analysis demonstrated that ponasterone A induction caused the expression of PE II-GrBa fusion proteins, and indirect immunofluorescence detected giant sized multinucleated cells, suggesting cytoskeletal and mitotic abnormalities as reported in our previous studies. Western blot, enzymatic activity assay and cell counting analysis indicated that two types of PE II-GrBa fusion proteins were capable of cleaving both endogenous and exogenous substrates of granzyme B, and inhibiting the growth of cells. The PE II (aa 280-358)-GrBa was shown to have higher serine protease activity and stronger growth inhibitory effect. Such inhibition was presumably associated with G2 arrest as determined by cell cycle analysis. These data prove that PE II-GrBa fusion proteins have cell inhibitory effect similar to GrBa, and that the shorter PE-derived peptide exerts less influence on GrBa activity. This study helps to optimize the construction of recombinant protein comprising translocating peptides and cytotoxic molecules for tumor cell killing.
ADP Ribose Transferases ; genetics ; pharmacology ; Bacterial Toxins ; genetics ; pharmacology ; Cell Proliferation ; drug effects ; Exotoxins ; genetics ; pharmacology ; Granzymes ; genetics ; pharmacology ; HeLa Cells ; Humans ; Recombinant Fusion Proteins ; pharmacology ; Virulence Factors ; genetics ; pharmacology

Fifteen known compounds were isolated from Swertia delavayi by silica gel, Sephadex LH-20 and Rp-18 column chromatographies. Based on extensive spectroscopic analysis (MS, 1H, 13C-NMR), their structures were identified aserythrocentaurin (1), erythrocentaurindimethylacetal (2), sweroside (3), swertiamarin (4), gentiopicroside (5), swertiakoside A (6), 2'-O-acetylswertiamarin (7), 4'-O-[(Z) -coumaroyl] swertiamarin (8), 1,5,8-trihydroxy-3-methoxyxanthone (9), 8-O-β-D-glucopyranosyl-1-hydroxy-2,3, 5-trimethoxyxanthone (10), 8-O-[β-D-xyl- opyranosyl-(1 --> 6)-β-D-glucopyranosyl]-7,8-dihydroxy-3-methoxyxanthone (11), isovitexin (12), β-sitosterol (13), daucosterol (14), and oleanolic acid (15). Among them, ten ones (14, 7-11, 13) were obtained from S. delavayi for the first time. The isolates were evaluated for their anti-HBV activities in HepG 2. 2. 15 cell line in vitro. The results showed that compound 1, 2, 6, 7, 9 and 12 exhibited significant inhibitory activity on HBV DNA replication with IC50 values from 0.05 to 1.46 mmol x L(-1).
Antiviral Agents ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Hepatitis B virus ; drug effects ; genetics ; Magnetic Resonance Imaging ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Swertia ; chemistry

OBJECTIVEAnalyze Naive and Mermory T cell subsets in HIV/AIDS patients and investigate their relationship with disease development.

METHODST cell subsets from 15 normal control subjects, 79 HIV/ AIDS patients were detected by FCM.

RESULTSWith diesase progression, CD4+ Naive cell counts and ratio was both decreased obviously (P < 0.001); CD4+ Tcm cell counts was significantly decreased (P < 0.001), CD4+ Tcm cell ratio was obviously higher (P = 0.002); CD4 TEM cell ratio was obviously increased( P < 0.001); CD8+ T Naive cell counts and ratio was also decreased obviously (P < 0.05); CD8+ T(CM), T(EM), T(EMRA) are not significantly different.

CONCLUSIONSThe peripheral lymphocyte subsets in HIV/AIDS patients changed obviously. The counts of naive T cell decreased, while the proportion of memory T cell increased significantly. It will help to understand pathogenesis of HIV.

Acquired Immunodeficiency Syndrome ; immunology ; virology ; Adult ; CD4 Lymphocyte Count ; Case-Control Studies ; Female ; HIV ; immunology ; Humans ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo investigate the effect of c-kit mutation on the prognosis of gastrointestinal stromal tumors.

METHODSA search of studies in PubMed and MedLine (from 1999 to 2008) was performed to assess the effect of c-kit mutation on the prognosis of gastrointestinal stromal tumors. The articles were retrieved with the entries of "gastrointestinal stromal tumors", "imatinib", "c-kit" and "mutation". A meta-analysis was performed to assess the data included.

RESULTSA total of 15 articles were collected in this analysis. No significant differences was found in incidence of mitoses (> 5/50 HPF) between the patients with wild type c-kit (wild type group) and the ones with mutated c-kit (mutation group) (P = 0.710); tumor recurrence and metastasis rate after surgery was significant higher in the mutation group than that in wild type group (P = 0.010); as for imatinib response with different c-kit mutation types, the results showed the incidence of clinical response (complete response + partial response) was significantly higher in mutation group than that in wild type group (P = 0.009), but the imatinib resistance rate was lower in mutation group (P = 0.000); three studies provided data for imatinib resistance with c-kit second mutations, the results showed the second mutations mainly focus on exon 13, 14, 17.

CONCLUSIONSC-kit mutation is related closely with the incidence of recurrence and metastasis in GIST after surgery. The mutations of c-kit influences the therapeutic effects of imatinib.

Antineoplastic Agents ; therapeutic use ; Benzamides ; Case-Control Studies ; Gastrointestinal Stromal Tumors ; drug therapy ; genetics ; Humans ; Imatinib Mesylate ; Mutation ; Piperazines ; therapeutic use ; Prognosis ; Proto-Oncogene Proteins c-kit ; genetics ; Pyrimidines ; therapeutic use