Human erythropoietin receptor (hEPOR) plays an important role in regulating the red blood cell production by promoting the proliferation and differentiation of RBC from erythroid precursors, hEpoR is a transmembrane protein, and its extrocellular domain (sEpoR) is of great importance in Epo signal transduction pathway. We cloned the gene of sEpoR by RT-PCR from the total RNA of human fetal liver and expressed it in E. coli after insertion of the gene in the expression vector pBV220. The cloned gene was confirmed by sequencing analysis and gene product was confirmed by both Western blot and its first 11 amino acid residues sequence of the N-terminal. In vitro bioassay showed that the purified gene product can repress the growth of TF-cells in the presence of Epo.

OBJECTIVETo explore the cure effects of Jiangu Fufang on osteoporotic model induced by ovariectomy.

METHODRats were ovariectomized and administered drugs for 3 monthes. Bone mineral density and biomechanics properties, histomorphometric analysis and biochemical index such as calcium, phosphorus, alkaline phosphatase were detected.

RESULTJiangu Fufang could significantly increase bone density and biomechanics properties. The level of calcium, phosphorus and alkaline phosphatase were restored by Jiangu Fufang. Jiangu Fufang could significantly increase area of bone trabecula, thickness of cortical bone and bone trabecula.

CONCLUSIONJiangu Fufang could cure osteoporosis through increasing bone mineral density, improving bone biomechanics properties, and effecting bone metabolism.

Alkaline Phosphatase ; blood ; Animals ; Biomechanical Phenomena ; Bone Density ; drug effects ; Calcium ; blood ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; therapeutic use ; Female ; Femur ; physiopathology ; Lumbar Vertebrae ; drug effects ; pathology ; physiopathology ; Osteoporosis ; blood ; drug therapy ; physiopathology ; Ovariectomy ; Phosphorus ; blood ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

AIMTo investigate the paclitaxel-loaded nanoparticles of poly(ethylene glycol)-b-poly(D,L-lactic acid) amphiphilic diblock copolymer (PMT).

METHODSPMT was prepared by solid dispersion technique. The average size and size distribution were determined by dynamic light scattering (DLS). The morphology was characterized by transmission electron microscopy (TEM) and 1HNMR. The influences of the copolymer molecular weight and the paclitaxel-fed amount on PMT were studied. Therapeutic effect of PMT was studied on Kunming mice liver cancer H22.

RESULTSPMT showed nanometer size and spherical morphology with core and shell. The sizes of PMT increased with increasing the molecular weight of the hydrophobic segment in PEDLLA or increasing the drug-loaded amount. The tumour inhibiting effect of PMT was similar with that of Taxol.

CONCLUSIONIt will provide an experiment basis for the development of new kind of intravenous administration of paclitaxel.

Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Lactic Acid ; Liver Neoplasms, Experimental ; pathology ; Mice ; Microspheres ; Nanotechnology ; Paclitaxel ; administration & dosage ; pharmacology ; Particle Size ; Polyethylene Glycols

OBJECTIVETo study the relationship between simian acquired immunodeficiency syndromn (SAIDS) and autoimmunity in simian immunodeficiency virus (SIV)-infected monkeys.

METHODSIndirect immunofluorescence assays were performed to detect plasma or serum autoantibodies in SIV-infected monkeys. The heart, liver, spleen, lung, kidney, and lymph node of BALB/c mice, a strain of endothelial cell ECV304, and granulocytes were used as target antigens. These results were compared with HE stained slides of SIV-infected monkeys.

RESULTSThe levels of various autoantibodies, including anti-lymphocyte autoantibodies, anti-endothelial cell autoantibodies, and anti-granulocyte antibodies, increased after SIV infection in monkeys. Moreover, pathological examinations showed injuries in the lymphoid tissue and vascular pathological changes in cerebral cortex, submucosa of gastrointestinal tract, interstitial capillaries of myocardium, nephron of the kidney, and sinusoid cell of liver.

CONCLUSIONThe increased autoantibodies and the pathological changes of tissues and organs confirm the existence of autoimmunity in SIV-infected monkeys.

Animals ; Autoantibodies ; blood ; Autoimmunity ; Endothelial Cells ; immunology ; Granulocytes ; immunology ; Lymphocytes ; immunology ; Mice ; Mice, Inbred BALB C ; Simian Acquired Immunodeficiency Syndrome ; immunology ; pathology ; Simian Immunodeficiency Virus

This study was aimed to investigate the relationship between endostatin and vascular cell adhesion molecule-1 (VCAM-1) expressions on bone marrow stromal cells (BMSC) in mice after bone marrow transplantation (BMT) and effect of ligustrazine on their expressions. The mice were randomly divided into 3 groups: normal group (without treatment), saline group (control of BMT) and ligustrazine group (BMT + ligustrazine). BMT mouse models were established. The normal group was not treated, the saline group was given normal saline (0.2 ml/mouse, twice a day) through gastric tube, while the ligustrazine group was given ligustrazine (0.2 ml/mouse, twice a day) also through gastric tube. On day 7, 14, 21 and 28 after BMT, mice were killed by euthanasia. The expression levels of endostatin and VCAM-1 in bone marrow stromal cells were detected by immunohistochemistry and RT-PCR analysis respectively. The results showed that the endostatin protein mainly expressed in nuclei of BMSCs, the VCAM-1 protein mainly expressed in plasma of BMSCs. On day 7, 14, 21 after BMT the expression levels of endostatin mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), while their expression levels in ligustrazine group were lower than that in saline group. On day 28 the expression levels in saline group returned to normal, while the expression levels in ligustrazine group not were normalized. On day 7, 14, 21 after BMT the expression levels of VCAM-1 mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), but their expression levels in ligustrazine group were significantly lighter than that in saline group (P < 0.01 or P < 0.05). On day 28 the VCAM-1 expression level in ligustrazine group returned to normal, while its expression level in saline group not were normalized. The difference between these two groups was significant (P < 0.01). Correlation analysis revealed that there was a negative correlation between endostatin and VCAM-1 expression in saline group, there was a positive correlation between endostatin and VCAM-1 expression in ligustrazine group. It is concluded that the endostatin can influence hematopoiesis in bone marrow by affecting VCAM-1 expression on BMSC and hindering connection between stromal cells and hematopoietic cells as well as extracellular stroma and hematopoietic cells, while ligustrazine can enhance the adhesion molecule expression on stromal cell surface of bone marrow in BMT-mice, accelerate the homing and proliferation of HSPC in bone marrow after BMT, meanwhile can promote the repair of bone marrow microenvironment, accelerate hematopoietic reconstitution of bone marrow after BMT through feedback regulation of endostatin expression of BMSC in BMT-mice.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Bone Marrow Transplantation ; Endostatins ; biosynthesis ; genetics ; Female ; Male ; Mice ; Mice, Inbred BALB C ; Pyrazines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Stromal Cells ; metabolism ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

BACKGROUNDAppropriate antimicrobial therapy of community-acquired pneumonia (CAP) is mainly based on the distribution of etiology and antimicrobial resistance of major pathogens. We performed a prospective observational study of adult with CAP in 36 hospitals in China.

METHODSEtiological pathogens were isolated in each of the centers, and all of the isolated pathogens were sent to Zhongshan Hospital for antimicrobial susceptibility tests using agar dilution.

RESULTSA total of 593 patients were enrolled in this study, and 242 strains of bacteria were isolated from 225 patients. Streptococcus pneumoniae (79/242, 32.6%) was the most frequently isolated pathogen, followed by Haemophilus influenzae (55/242, 22.7%) and Klebsiella pneumoniae (25/242, 10.3%). Totally 527 patients underwent serological tests for atypical pathogens; Mycoplasma pneumoniae and Chlamydia pneumoniae infections were identified in 205 (38.9%) and 60 (11.4%) patients respectively. Legionella pneumophila infections were identified in 4.0% (13/324) of patients. The non-susceptibility rate of isolated Streptococcus pneumoniae to erythromycin and penicillin was 63.2% and 19.1% respectively. Six patients died from the disease, the 30-day mortality rate was 1.1% (6/533).

CONCLUSIONSThe top three bacteria responsible for CAP in Chinese adults were Streptococcus pneumonia, Haemophilus influenza and Klebsiella pneumonia. There was also a high prevalence of atypical pathogens and mixed pathogens. The resistance rates of the major isolated pathogens were relatively low except for the high prevalence of macrolide resistance in Streptococcus pneumoniae.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bacteria ; drug effects ; isolation & purification ; pathogenicity ; China ; epidemiology ; Colony Count, Microbial ; Community-Acquired Infections ; drug therapy ; etiology ; microbiology ; mortality ; Drug Resistance, Bacterial ; Female ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Pneumonia, Bacterial ; drug therapy ; etiology ; microbiology ; mortality ; Prospective Studies

To study the alteration of the cell cycle during the differentiation of human myeloid leukemia cell line HL-60 induced with all trans-retinoic acid (RA), the flow cytometry was used to assay the various phases of cell cycle in HL-60 cells treated with RA. The results showed: (1) S + G(2)/M phase proportion kept relative invariability during the 48 hours incubation of HL-60 cells with RA, however, the proportion alteration of S-phase cells was associated with the RA concentrations. At 10(-6) mol/L RA, the proportion of S-phase cells appeared a temporarily increasing peak followed by persistent decrease of S-phase proportion. At 10(-5) mol/L RA, S-phase cell proportion only appeared the persistent decreasing tendency. (2) Re-culture of HL-60 cells without RA showed the decrease of S + G(2)/M and S-phase cells was associated with the increase of differentiated cells, but not all HL-60 cells were triggered into differentiation at the same time. Once the cells start to differentiate, even if there is no RA presence, the HL-60 cells were still differentiated until maturation. In conclusion, HL-60 cells are able to differentate and maturate after exposure to RA for a period of time. S-phase proportion is related to the concentrations of RA. Once the cells start to differentiate, even if there is no RA presence, the HL-60 cells still differentiatiated until maturation.

OBJECTIVETo explore the characteristics of the whole genome of the influenza H1N1 virus of the mild and severe cases in Beijing.

METHODSA total of 21 samples of throat swabs were collected from surveillance-designated hospitals between June and December in 2009, including 10 severe cases (4 death cases) and 11 mild cases. RNA of the virus were extracted,and the amplified primers of the whole genome were designed.Reverse transcription and PCR were performed to the RNA and then the PCR product was sequenced by software to analyze the evolution of the viral genes and the variation of the amino acids.

RESULTSCompared with the reference vaccine strain A/California/07/2009 (H1N1), the genetic nucleotide homology in the eight segments of the pandemic H1N1 virus in Beijing in 2009 was higher than 99%, without significant variation. Among them,the genetic distance of hemagglutinin (HA), neuraminidase (NA) and nucleoprotein (NP) was comparatively far, separately 0.0050, 0.0040 and 0.0040.The gene of HA, P83S, the gene of NA, N248D, the gene of polymerase (PA), P224S and the gene of NP, V100I and L122Q were found to mutate in all the samples. Genes of HA, NA, NP, PA, PB 2 and nonstructural protein (NS1) in severe cases showed obviously clustered evolution. The mutation of gene S128P and S203T of HA, gene R269R and D547E of PA, gene T588I of PB 2 and gene I123V of NS mainly happened in severe cases, separately counting 6, 9, 6, 7, 9 and 6 cases. The relevance between the mutation happened in S203T of HA, R269K and D547E of PA and the severeness of the cases showed statistical significance (P < 0.05). The mutations of HA gene were mainly on the Ca and Cb antigene domains. No drug resistant mutation was found on NA gene but happened on matrix protein 2 (M2 gene). None of the mutations were found on the virulence related genes.

CONCLUSIONA high homology was found between the pandemic H1N1 virus in Beijing in 2009 and the reference vaccine strain A/California/07/2009(H1N1). Mutational sites related with the severe and fatal cases were found, but not the virulence related mutation.

Base Sequence ; China ; epidemiology ; Genes, Viral ; Genetic Variation ; Genome, Viral ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; Influenza, Human ; epidemiology ; virology ; Neuraminidase ; genetics ; RNA-Binding Proteins ; genetics ; Viral Core Proteins ; genetics

OBJECTIVETo explore the risk factors for hepatoblastoma.

METHODSA case-cohort study using Logistic regression multiple variables analysis of medical record data sets was conducted to examine infant and perinatal risk factors for hepatoblastoma.

RESULTSBirth weight less than 1,000 g was associated with a strongly increased risk of hepatoblastoma (odds risk, OR = 26.0, 95% confidence interval, CI: 14.0 to 65.7). After adjustment of birth weight, a moderately increased risk of hepatoblastoma was found for older maternal age ( > 35 years vs. 20 to 34 years: OR = 2.6, 95% CI: 0.9 to 5.9), maternal smoking (OR = 2.9, 95% CI: 1.1 to 4.2) and higher maternal pregnancy body mass index (OR = 3.2, 95% CI: 1.0 to 6.7).

CONCLUSIONVery low birth weight and maternal characteristics including overweight, smoking are associated with hepatoblastoma risk.

Case-Control Studies ; Child ; Child, Preschool ; Confidence Intervals ; Female ; Follow-Up Studies ; Hepatoblastoma ; epidemiology ; etiology ; prevention & control ; Humans ; Infant ; Infant, Newborn ; Infant, Very Low Birth Weight ; Liver Neoplasms ; epidemiology ; etiology ; prevention & control ; Male ; Overweight ; Pregnancy ; Retrospective Studies ; Risk Factors ; Smoking ; adverse effects

Objective:To screen and identify the protein that interacts with the adhesion protein of Mycoplasma genitalium (MgPa)from T7-phage display cDNA library of human uroepithelial cells(SV-HUC-1).Methods:Recombinant adhesion protein of My-coplasma genitalium(rMgPa)was used as target molecule to biopan the T7 phage display cDNA library of SV-HUC-1 cell,the selected positive clones were analysed using DNA sequencing and BLAST analysis and identified by means of indirect ELISA,Dot immunoblot and Far-western blot.Results:After four rounds of biopanning,positive phages were obviously enriched.According to the results of DNA sequencing and BLAST analysis,the selected randomly 32 positive clones included 7 kinds different sequences,of which the number of RPL35 repeats was the most.The results of indirect ELISA,Dot immunoblot and Far-western blot showed that 7 representative phages could bind specifically with rMgPa.Conclusion:60S ribosomal protein L35(RPL35) may be the interacting protein of MgPa,which lays the experimental foundation for understanding the function of MgPa and the pathogenesis of Mycoplasma genitalium.