Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; statistics & numerical data ; Pesticides ; Rural Population ; statistics & numerical data ; Sampling Studies ; Young Adult

Aged ; Coal Mining ; Electrocardiography ; Heart Rate ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; complications ; physiopathology ; Tachycardia ; etiology

Objective:To establish the quality standard for Huashengping capsules. Methods: Milkvetch Root, Hedyotis diffusa Willd and Salvia miltiorrhiza were identified by TLC. The content of astragaloside A was detected by HPLC. The column was Kormasil C18(250 mm ×4.6 mm, 5 μm) and the flow rate was 1.0 ml·min-1. The mobile phases was a mixture of acetonitrile-water (32∶68 ) . The detection wavelength was 203 nm. The column temperature was 25℃ and the sample size was 20μl. Results:The TLC chro-matography spots were clear. Astragaloside A was linear within the range of 2. 000-10. 000 μg(r=0. 999 6) and the average recovery was 100. 8%(RSD=1. 9%,n=6). Conclusion:The method is simple, accurate and reliable, which can be used in the quality con-trol of Huashengping capsules.

To assay the influence of dendritic cells(DCs)on the function of anti-infective immunity to Penicillium marneffei.DCs were generated from peripheral blood mononuclear cells(PBMC)and pulsed with Penicillium marneffei yeasts.DCs morphology was observed by the inverted microscope and cell surface markers of DCs were analyzed by flow cytometry.The concentrations of IL-12p70 were detected by ELISA. Mixed lymphocyte reaction was performed to assay the proliferation of T cells.The mRNA of CCR7 and CXCR4 were detected by the Real-time PCR quantifications.The acquired DCs exhibited irregular appearance and numerous long dendrites under light microscope.DCs and Penicillium marneffei yeasts were co-cultured for 24 h,numerous yeasts were observed inside the cells;an enhanced expression of the cell sur-face markers CD86、CD83、HLA-DR and CD40 were demonstrated;the expression of CCR7 and CXCR4 mRNA were also increased;the improved proliferation of T cells were observed in the mixed lymphocyte reaction.Yeasts-pulsed DCs secreted more IL-12p70 than that of non-pulsed,but less than that of LPS-pulsed DCs.DCs can engulf the Penicillium marneffei yeasts.When pulsed with Penicillium marneffei yeasts,DCs improved their expression of the co-stimulatory molecules and chemokine receptor CCR7、CXCR4,enhanced their capacity to process antigen.DCs play an important role in host defense against Penicillium marneffei infection.But the low level of the IL-12p70 production may lead to deficiency in the cell-mediated immunity against Penicillium marneffei.

OBJECTIVEDNAzyme/Deoxyribozyme is another novel molecular biological tool following the ribozyme. DNAzyme consists of a 15-nucleotide (nt) internal loop as its catalytic domain and two flanking substrate-recognition domains of 7 to 8 nt each which is complementary to substrate. The RNA substrate is cleaved at a particular phosphodiester located between an unpaired purine and a paired pyrimidine residue. DNAzyme has been applied in fields such as viral infectious disease, tumor, cardiovascular disease and genetic disease. But there is no report about using DNAzyme for anti-respiratory syncytial virus purpose. To observe the inhibitory effects of RNA-cleaving DNAzymes on respiratory syncytial virus (RSV) replication in cultured cells.

METHODSAnti-RSV RNA-cleaving DNAzyme DZ604 was designed to target the RSV genome at the start of the NS2 gene in an effort to inhibit the RNA replication. Microscope and electron microscope were used to observe the effects of anti-RSV genomic RNA DNAzyme on cytopathogenic effect (CPE) and ultrastructural change of 9HTE cell induced by RSV infection. Viral plaque forming reduction assay and MTT assay were used to detect the anti-RSV activity and protective function for RSV infected 9HTE cells of DNAzyme.

RESULTSAnti-RSV genomic RNA DNAzyme (DZ604) significantly improved CPE of RSV-infected 9HTE cells. The time to appearance of CPE and of total CPE was delayed by using DZ604 in a dose-dependent manner. At a 5 micro mol/L concentration of DZ604, CPE of 9HTE cells induced by RSV infection at 10 and 1 multiplicity of infection (MOI) was not improved. At smaller MOI (0.1, 0.01, 0.001, 0.0001) of RSV infection, CPE of 9HTE cells was significantly lightened by DZ604 at the same concentration. DZ604 also significantly improved ultrastructural change of 9HTE cells at early stage of RSV infection. Reduction in RSV yield was 85.56% and 8.33% at concentrations of 5 micro mol/L and 0.25 micro mol/L of DZ604. DZ604 inhibited RSV yield in a dose-dependent manner (P < 0.05). Non-specific DNAzyme did not have anti-RSV activity (P > 0.05).

CONCLUSIONAnti-RSV genomic RNA DNAzyme designed and synthesized in our laboratory was capable of inhibiting respiratory syncytial virus replication specifically in cultured cells. Our data indicated that DNAzymes could be useful for the prevention against respiratory syncytial virus infection.

Animals ; Cell Line ; Cercopithecus aethiops ; DNA, Catalytic ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Microscopy, Electron ; Respiratory Mucosa ; cytology ; ultrastructure ; virology ; Respiratory Syncytial Viruses ; drug effects ; genetics ; physiology ; Vero Cells ; Virus Replication ; drug effects

OBJECTIVETo investigate the relationship between various Chinese medicine (CM) types and T-cell subsets (CD4(+) and CD8(+)) in the colonic mucous membranes of patients with ulcerative colitis (UC).

METHODSFifty UC patients were enrolled, after differentiation into four types by CM syndromes, i.e., the internal heat-damp accumulation type (IHDA), the qi-stagnancy with blood stasis type (QSBS), the Pi-Shen yang-deficiency type (PSYD) and the yin-blood deficiency type (YBD). From every patient, 3-5 pieces of intestinal mucous membrane tissues were taken through colonoscopy to determine the levels of the T-cell subsets (CD4(+) and CD8(+)) using immunohistochemical method. The results were compared with those in the normal control.

RESULTSThe level of CD8(+)increased and the ratio of CD4(+)/CD8(+)decreased mainly in colonic mucous membranous tissues in UC patients. The level of CD4(+)decreased significantly in IHDA types (P<0.01), but decreased only slightly in the PSYD, QSBS and YBD types. CD8(+)increased significantly in the IHDA types (P<0.01), but only slightly in the other three types.

CONCLUSIONThe IHDA types of UC are closely related with T-cell subsets. The difference of T-cell subsets in various IHDA types of UC patients has provided a theoretical basis for syndrome differentiation in the CM typing of UC.

Adolescent ; Adult ; Aged ; Biopsy ; Blood Circulation ; CD4-CD8 Ratio ; Colitis, Ulcerative ; immunology ; pathology ; Colon ; drug effects ; immunology ; pathology ; Colonoscopy ; Female ; Humans ; Immunohistochemistry ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Qi ; T-Lymphocyte Subsets ; drug effects ; Yang Deficiency ; immunology ; pathology ; Yin Deficiency ; immunology ; pathology ; Young Adult

The monocot mannose-binding lectin can inhibit HIV from infecting the target cells. The total RNA of Zephyranthes grandiflora was extracted and reversely transcribed into cDNA. Degenerate primers were designed based on the conserved regions of other monocot mannose-binding agglutinins by homology alignment. The 694bp full-length cDNA of Zephyranthes grandiflora agglutinin (ZGA) was cloned by RT-PCR, 3' and 5' RACE (rapid amplification of cDNA ends). The start codon and stop codon of ZGA were at 37-39bp and 529-531bp respectively. The NCBI Blast analysis result showed that ZGA gene encoded a protein precursor with signal peptide, mature protein and C-terminal cleavage sequence. The mature ZGA protein contained 106 amino acids residues and its molecular weight was 11.6KD. The percentages of identity of the deduced mature ZGA protein with those of Galanthus nivalis agglutinin, Narcissus hybrid cultivar agglutinin, Lycoris radiate agglutinin and Clivia miniata agglutinin were 71.8%, 81%, 81.8% and 84.5%, respectively. Blocks analysis revealed that ZGA had three functional domains and three mannose-binding boxes (QDNY).
Agglutinins ; genetics ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Liliaceae ; genetics ; Mannose-Binding Lectin ; genetics ; Molecular Sequence Data ; Sequence Analysis, DNA

OBJECTIVETo explore the feasibility of regulated expansion and committed differentiation potential of JAK2 gene modified hematopoietic stem/progenitor cells in vitro.

METHODSA murine stem cell virus (MSCV) based retroviral vector MGI-F2Jak2, which encodes a green fluorescent protein (GFP) and a fusion protein containing two copies of modified FK506 binding protein (F36v) linked tyrosine kinase JAK2 was cloned. F36v served as a high-affinity binding site for dimerizer AP20187. GpE + 86 packaging cell was transfected with this vector. Bone marrow cells from C57BL/6 mice were transduced by co-cultured with irradiated (1500 cGy) GpE + 86 producer clone for 48 h. Transduced marrow cells were expanded in X-VIVO 15 and divided into four groups as follows: (1) control group; (2) AP20187 alone group; (3) SCF alone group and (4) AP20187 + SCF group. The phenotypes of the expanded cells were analyzed by directly phycoerythrin-labeled anti-Sca1, c-kit, CD(34), Gr1, CD(11b), TER119, CD(41), B220 and CD(3) monoclonal antibodies for flow cytometry. Committed differentiation, progenitor colony assay and spleen colony forming units (CFU-S) were further evaluated.

RESULTSA significant sustained outgrowth of transduced marrow cells was obtained only in the AP20187 + SCF group. Cells expanded up to 10(14)-fold after 80 days culture. The doubling time was about 30 hs. The phenotypes of the expanded cells were homogeneously strong positive for CD(34), c-kit and Sca1, while were almost negative for Gr1, CD(11b), TER119, CD(41), B220 and CD(3). Functional assay demonstrated that the expanded cells had multipotential to differentiate into granulocyte, macrophage, erythrocyte, or B-cells under different cytokines combinations. A prominent megakaryocytic differentiation was observed when cultured with SCF/Tpo/IL-11 combination. The expanded cells were also capable of forming BFU-E, CFU-GM and CFU-Mix in methylcellulose colony assay. The expanded cells over three months could still form CFU-S.

CONCLUSIONSAP20187 combined SCF mediated activation of JAK2 signaling domain can dramatically expand hematopoietic stem/progenitor cells, and the expanded cells can be regulated and committed to differentiate into multilineage cells. This system may provide important insights into stem cell biology and may be promising for gene and cell therapy.

Animals ; Cell Differentiation ; Female ; Hematopoietic Stem Cells ; cytology ; Janus Kinase 2 ; Mice ; Mice, Inbred C57BL ; Protein-Tyrosine Kinases ; genetics ; physiology ; Proto-Oncogene Proteins ; Tacrolimus ; analogs & derivatives ; pharmacology ; Transfection

The purpose of this study was to investigate the effect of bisphosphonate combined with chemotherapy on bone mineral density (BMD) of patients with multiple myeloma (MM) and analyse its value of BMD detection in clinic of these patients. 53 MM cases were enrolled in this study, including 33 newly diagnosed, 10 refractory/relapsed and 10 stable cases. They were divided randomly into two groups, 33 MM cases were treated with bisphosphonates combined with chemotherapy and 20 MM cases were treated with chemotherapy alone. The chemotherapy schedules for all patients were same. BMD was tested using the dual energy X-ray absorptiometry at 2 time points, i.e. pretreatment (basal level) and 12 months after treatment with chemotherapy and bisphosphonates. Comparisons was performed with t tests using SPSS 11.0 software. The results indicated that there was minor difference between 2 groups for BMD scores of whole body and lumbar vertebra (L1-L4), but no difference for scores of the near-end of left femur. After treatment for 12 months, all BMD scores (whole body, lumbar vertebra and the near-end of left femur) increased significantly in the bisphosphonate combined with chemotherapy group (P < 0.05). In contrast, only minor changes were seen in chemotherapy alone group. It is concluded that the bisphosphonate combined with chemotherapy has displayed promotive effect on BMD of MM patients, the detection of BMD is sensitive and special method for monitoring therapeutic effect of bisphosphonate in MM patients, thus has useful value in clinic of these patients.
Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Bone Density ; drug effects ; Diphosphonates ; administration & dosage ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; metabolism ; pathology

OBJECTIVETo study the methylation status of p73 gene promoter in patients with myelodysplastic syndrome (MDS) and explore its significance with clinical prognosis.

METHODSMethylation of p73 promoter was detected in bone marrow cells from 135 MDS patients and 13 healthy controls by methylation-specific PCR (MSP). The results of MSP were confirmed by bisulfite sequencing. The expression of p73 mRNA was detected by real-time quantitative PCR. Primary bone marrow cells from MDS patients were treated with decitabine, the changes of p73 methylation status and p73 mRNA expression were measured. The role of p73 methylation in the prognosis of MDS and the correlated clinical data were explored.

RESULTSp73 hypermethylation was present in 37.04% of MDS cases and patients with high risk MDS (RAEB-1 and RAEB-2) exhibited a significantly higher frequency of p73 methylation than that of low risk MDS (58.8% vs 29.7%, P = 0.002). The expression of p73 mRNA in the methylated group was decreased compared to that of the unmethylated group (P = 0.032). Decitabine treatment decreased the level of p73 methylation and increased the level of p73 transcripts. Patients with p73 methylation progressed rapidly to AML (P < 0.001) and had shorter survival (P = 0.002) than those who did not have p73 methylation. In the multivariate Cox regression model, BM blast and p73 methylation status emerged as independent prognostic factor for overall survival and leukemia free survival.

CONCLUSIONp73 gene methylation is common in patients with MDS and may indicate poor prognosis. p73 may be a therapeutic target in MDS.

Aged ; Case-Control Studies ; DNA Methylation ; DNA-Binding Proteins ; genetics ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Nuclear Proteins ; genetics ; Prognosis ; Promoter Regions, Genetic ; Tumor Protein p73 ; Tumor Suppressor Proteins ; genetics