Objective To study the treatment of deep vein thrombosis (DVT) of lower limbs. Methods A series of 82 patients with DVT treated in this hospital from July 1995 to July 2002 were retrospectively analyzed. Sixty-two patients underwent conservative therapy while 20 patients underwent thrombectomy. Results Oedema on diseased limbs subsided in varying degrees in all the patients. However, complete re-perfusion was achieved in only 15 patients, and partial re-perfusion in 9 patients. In the rest of 58 patients, thrombosis was unimproved or recurred, leaving behind the post-thrombotic syndrome. Conclusions Early treatment for acute DVT, conservative therapy or surgical intervention, is effective. Surgery is advisable in most early central type or mixed type patients, whereas conservative therapy in late central type or peripheral type patients. The intravenous interventional therapy is a relatively novel and favorable vascular technique.

To provide accurate information on geographic distribution of crude drug Sailonggu in the plateau, we identified zokor species (Eospalax spp.) in Qinghai-Tibet Plateau using molecular methods. Based on the mitochondrial cytochrome B (cytb) gene sequences, we then extracted haplotypes from these sequences and reconstructed phylogenetic trees for the haplotypes using both maximum likelihood (ML) and Bayesian inference (BI) methods. Based on the trees, the species of each sample were determined. Five hundred and three samples from 35 populations were sequenced and their whole cytb sequences (1140 bp) were obtained. From these sequences 150 haplotypes were detected, in which, 126 were Eospalax baileyi, 20 were E. cansus, and 4 were E. smithi of the 35 populations, 28 were E. baileyi type, 5 were E. cansus type, and the remaining 2 were mixed of E. baileyi + E. cansus (DT2) and E. baileyi + E. smithi (ZN3). The results showed that, the regions around the Qinghai lake and near the upper stream of Yellow River started at Guide could be viewed as the producing area of authentic Sailonggu, and also, the cytb gene is a powerful molecular marker to determine the species of zokors as well as for the authentication of geographic distribution of Sailonggu.
Animals ; Bone and Bones ; metabolism ; Haplotypes ; Medicine, Tibetan Traditional ; Phylogeny ; Rodentia ; classification ; genetics

OBJECTIVETo study the serum level of interleukin-17A (IL-17A) and its expressions in the lung, spleen and thymus in asthmatic mice.

METHODSIn 14 normal BALB/c female mice and 14 asthmatic mice, the changes in the airway pathology and the cell proportion in the bronchoalveolar lavage fluid (BALF) were observed. The serum level of IL-17A and IL-17A expressions in the tissue homogenates of the lung, spleen and thymus of the mice were detected by sandwich enzyme-linked immunosorbent assay (ELISA).

RESULTSThe airway inflammation in the asthmatic mice was characterized mainly by eosinophil and neutrophil infiltration, which was not observed in the normal control group. Serum IL-17A levels and IL-17A expressions in the lung, spleen and thymus of the asthmatic mice were significantly higher than those in the normal control group (P<0.01). In the asthmatic mice, IL-17A expression in the lung tissues was positively correlated with the percentages of neutrophils (r=0.693, P=0.040) and eosinophils (r=0.733, P=0.030) in the BALF.

CONCLUSIONIL-17A is highly expressed in the serum, lung, spleen and thymus of asthmatic mice. IL-17A may be one of the major cytokines involved in exacerbation of bronchial asthma, and is probably associated with the recruitment of neutrophils and eosinophils into the airways.

Animals ; Asthma ; blood ; etiology ; metabolism ; Bronchoalveolar Lavage Fluid ; chemistry ; Female ; Interleukin-17 ; blood ; metabolism ; Mice ; Mice, Inbred BALB C ; Random Allocation

OBJECTIVE: To investigate the effect of oxidative stress on the accumulation of heat shock protein 70 (HSP70) within C2C12 myogenic cells. METHODS: Heat shock response (42 degrees C for 1 h and recovery for 12 h at 37 degrees C) was used to induce the expression of heat shock protein 70. We constructed a recombinant plasmid of HSP70 with enhanced green fluorescent protein (EGFP). After being transfected transiently into C2C12 cells, immunoblotting was used to detect the expression of HSP70 induced by heat shock response and transfection. Immunocytochemistry, fluorescent microscopy and immunoblotting were used to detect the translocation of HSP70. RESULTS: Immunoblotting showed that the overexpression of HSP70 was induced by heat shock response and transient transfenction. HSP70 localized within the cytoplasm of the normal cells, but HSP70 translocated from the cytoplasm to the nucleus and the nucleolus at 1 h after the treatment of oxidative stress (0.5 mmol/L H2O2) by using immunocytochemistry, fluorescent microscopy and immunoblotting for cellular partial proteins. CONCLUSION Oxidative stress may induce the accumulation of heat shock protein 70 within the nucleolus.
Cell Nucleolus ; metabolism ; Cells, Cultured ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Myoblasts ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Oxidative Stress ; physiology

OBJECTIVETo evaluate the accuracy of sentinel lymph node mapping(SLM) in patients with rectal cancer by single-photon emission computed tomography (SPECT-CT) lymphoscintigraphy and carbon nanoparticles suspension injection.

METHODSTwelve patients with clinical T(1-2)N(0)M(0) rectal cancer were selected and locally injected with technetium-(99m)sulfur-colloid and carbon nanoparticles suspension by endoscope one day before surgery, followed by SPECT-CT scanning 1, 3 and 5 hours later. Radioactive isotope(RI) uptake of each sentinel node(SN) basin with location preoperatively determined by SPECT-CT was postoperatively calculated using gamma probe. Nodes with the highest RI uptake, the number of which was also pre-determined by SPECT-CT, was defined as SNs. Immunohistochemical cytokeratin staining was performed for all the SNs and non-SNs.

RESULTSThe rate of sentinel node detection was 91.7%(11/12) with at least one SN(1-3) per patient. Ten cases showed metastasis-negative in SNs as well as all the resected regional nodes by immunohistochemical cytokeratin staining. Only one patient had positive nodes in both SN and non-SNs. The accuracy of SLM was 100%.

CONCLUSIONSPECT-CT lymphoscintigraphy and carbon nanoparticles suspension injection can effectively detect the anatomic location and number of sentinel nodes, and improve the accuracy of SLM for rectal cancer.

Adult ; Aged ; Carbon ; Female ; Humans ; Male ; Middle Aged ; Nanostructures ; Rectal Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Sentinel Lymph Node Biopsy ; methods ; Tomography, Emission-Computed, Single-Photon ; methods ; Tomography, X-Ray Computed ; methods

OBJECTIVETo construct different mutants of human p53 for expression in eukaryotic cells and investigate the effects of these mutants on stress-induced cell apoptosis.

METHODSHuman p53 cDNA was amplified by PCR and cloned into pcDNA3/HA vector following the routine procedures. The Ser15 and Ser46 of p53 were mutated to Ala and identified by enzyme digestion and PCR, and these mutants were expressed in NIH3T3 cells and detected by Western blotting. After transfection with the plasmids of different p53 mutants, the NIH3T3 cells were double-stained with AnnexinV-FITC and propidium iodide for apoptotic analysis using flow cytometry.

RESULTSThe recombinant plasmids of HA-tagged wild-type p53, HA-p53(WT), and its mutants, HA-p53(S15A) and HA-p53(S46A), were successfully constructed and expressed efficiently in NIH3T3 cells. The apoptotic ratio of p53(WT)-transfected cells induced by arsenite increased and that of p53(S15A)-transfected cells decreased significantly after arsenite stimulation, but no significant changes occurred in the apoptosis of p53(S46A)-transfected cells.

CONCLUSIONThe phosphorylation on Ser15 of p53 plays an important role in mediating arsenite-induced cell apoptosis.

Animals ; Apoptosis ; drug effects ; Arsenites ; pharmacology ; Base Sequence ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Humans ; Mice ; Molecular Sequence Data ; Mutation ; NIH 3T3 Cells ; Phosphorylation ; Transfection ; Tumor Suppressor Protein p53 ; genetics ; metabolism

Adult ; Base Sequence ; Carcinoma, Hepatocellular ; metabolism ; Female ; Humans ; Insulin-Like Growth Factor II ; biosynthesis ; genetics ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics ; Sequence Analysis, DNA

OBJECTIVETo investigate the regulatory effect of Jingang Jiangu pill (see text, JGJG) on expression of integrin in ovariectomized rats.

METHODSFifty ovariectomized 10 months old female rats were randomly divided into 5 groups: Fushanmei group (FSM), Jingang Jiangu pill (see text) group (JGJG), Gusongbao granule group (GSB), Model group (OVX), Sham group. After ovariectomized,the rats were raised in the same environment for 13 weeks. The rats in JGJG group took 0.13 g JGJG pill orally each day for each rat; the rats in GSB group took 0.86 g GSB granule orally each day for each rat; the rats in FSM group took 0.28 mg FSM orally each day for each rat; and the rats in OVX and sham groups took sodium. The treatment duration of rats in above 5 groups was 13 weeks. Bone mineral density (BMD) and the expression of integrin beta1 and alphavbeta3 were detected in each group after the treatment. RESYKTS: The BMD and the expression of integrin beta1 in FSM group, JGJG group and GSB group improved obviously than that of OVX group. There were statistical difference between these groups (P<0.05). The expression of integrin alphavbeta3 of the three treating groups significantly depressed.

CONCLUSIONThe JGJG pill improves BMD and express of integrin beta1, in ovariectomized rats and reduces express of integrin alphavbeta3 through the regulation of the coupling of osteoblasts and osteoclasts.

Animals ; Bone Density ; Disease Models, Animal ; Female ; Integrin alphaVbeta3 ; analysis ; Integrin beta1 ; analysis ; Medicine, Chinese Traditional ; Osteoporosis ; drug therapy ; metabolism ; Ovariectomy ; Rats ; Rats, Wistar

OBJECTIVETo construct pNTAP-MK2 eukaryotic expression plasmid and establish a HEK293 cell line stably expressing tandem affinity purification (TAP)-tagged MK2.

METHODSThe MK2-encoding region was subcloned into the vector pNTAP to construct the recombinant plasmid pNTAP-MK2, which was subsequently transformed into DH5 alpha.E.coli. After identification by PCR, digestion with restriction endonuclease and sequencing, the recombinant expression plasmid was transfected into HEK293 cells via liposome, and the cell line with stable expression of exogenous TAP tag-MK2 gene was selected by antibiotic G418. The expression and localization of the fusion protein TAP tag-MK2 were detected by Western blotting and immunofluorescence assay.

RESULTSThe results of PCR, restriction endonuclease digestion and sequencing all confirmed the correct construction of the recombinant eukaryotic expression plasmid pNTAP-MK2. Western blotting showed that the recombinant plasmid was expressed stably in HEK293 cells after transfection with G418 selection. Immunofluorescence assay identified the expression product TAP tag-MK2 mainly in the cell nuclei.

CONCLUSIONThe eukaryotic expression vector pNTAP-MK2 has been successfully constructed, and in the established cell line with stable expression of TAP tag-MK2, TAP tag does not influence the localization of exogenous MK2.

Gene Expression ; Genetic Vectors ; HEK293 Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Plasmids ; Protein-Serine-Threonine Kinases ; genetics

OBJECTIVE: To explore the role of cell-surface nucleolin in lipopolysaccharide (LPS)-stimulated expression and secretion of TNF-alpha and IL-1beta in human THP-1 monocytes. METHODS: Immuno-fluorescence assay and Western blot were used to identify the expression of nucleolin on the surface of THP-1 monocytes. Inactivation of nucleolin was induced by anti-nucleolin monoclonal antibody blockage, and the expression and secretion of TNF-alpha and IL-1beta were observed by using reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immuno-sorbent assay (ELISA)respectively in LPS-mediated human THP-1 monocyte inflammatory model. RESULTS: Immuno-fluorescence showed that nucleolin was localized on the cell surface of THP-1 monocytes as indicated by dotted red fluorescence. Western blot assay indicated that nucleolin existed in the cell membrane fractions. RT-PCR assay showed that the expressions of TNF-alpha and IL-1beta mRNA significantly increased at 2 h and 3 h after the treatment with 1000 microg/L LPS. After 1 h pretreatment with anti-nucleolin antibody, the levels of TNF-alpha and IL-1beta mRNA decreased compared with an anti-nucleolin antibody untreated group and an irrelevant IgG+LPS group (P<0.05). ELISA assay showed that the pretreatment with anti-nucleolin antibody inhibited significantly the secretion of LPS-induced levels of TNF-alpha and IL-1beta after 4, 12 and 24 h treatment with 1000 microg/L LPS. CONCLUSION Nucleolin expresses on the cell surface of THP-1 monocytes and involves in the LPS-mediated expression and secretion of TNF-alpha and IL-1beta.
Cell Line ; Cell Membrane ; metabolism ; Humans ; Interleukin-1beta ; biosynthesis ; metabolism ; Lipopolysaccharides ; pharmacology ; Monocytes ; cytology ; metabolism ; Phosphoproteins ; metabolism ; physiology ; RNA-Binding Proteins ; metabolism ; physiology ; Tumor Necrosis Factor-alpha ; biosynthesis ; metabolism