Objective To investigate the effect of cobalt chloride on tumor cell proliferation and apoptosis in vitro, to explore the reasonable strategy of chemical hypoxia induced by CoCl2. Methods Two tumor cell lines (A549 and HeLa) were respectively exposed to CoCl2(0.05～2 mmol/L) for different time period (4～48 h), cells viability、proliferation and apoptosis were maesured by MTT and FCM methods. HIF-1? and related apoptosis proteins expression were detected by Western blot. Results Cells viability was weakly changed in low concentration(≤200 ?mol/L)of CoCl2 within 24 h. However, higher dose or prolonged challenge of CoCl2 significantly decreased cell survival rate (P

ObjectiveTo observe the influence of polyethylene glycol-conjugated hemoglobin (PEG-Hb) solution combined with cisplatin on the expression of erythropoietin/erythropoietin receptor (EPO/EPOR) and tumor angiogenesis in cancer treatment.MethodsHeLa cells were injected subcutaneously into the right oxter of 3-4 weeks old BALB/c nude mice to establish cervical tumor xenograft model.Then animals were randomly assigned to 4 groups (n=10) and treated respectively:group 1(control); group 2,cisplatin (5 mg/kg); group 3,PEG-Hb (0.6 g/kg); group 4 cisplatin (5 mg/kg) plus PEG-Hb (0.6 g/kg).The volume oftumors in each groups were measured in 4 weeks treatment period.Efficacy was measured as percent tumor growth inhibition (TGI) relative to salinetreated group.CD31 was detected by immunohistochemistry and its expression was identified as microvascular density (MVD).Expressions of hypoxia inducible factor-1α(HIF-1α) and EPO/EPOR in tumor tissues were analyzed by irnmunohistochemistry.EPOR protein level was tested by western blot.ELISA method was used in measuring EPO concentration in serum.ResultsTumor volume was significantly decreased in group 4 compared with other groups.Immunoreactivity data demonstrated lower expression of CD31,HIF-1α and EPO/EPOR in group 4.The expression of EPOR in the endothelial cells was also significantly decreased in group 4.Western-blot data indicated lower EPOR protein level in group 4.Serum level of EPO was also decreased in group 4.ConclusionPEG-Hb plus cisplatin is benefit to tumor tissue oxygenation,therefore it can inhibit the tumor angiogenesis and down regulate the erythropoietin/erythropoietin receptor system.The result can provide more evidence for the enhanced sensitivity effect of the artificial oxygen carrier in cancer therapy.

0.05).92.7%of the objects knew that passive smoking was harmful.71.9%knew that passive smoking made people suffering from cardiopathy more possibly.74.9%knew that the wife whose husband smoked were easier to catch lung cancer.And 84.4%knew that the child whose parent s smoked more possibly took asthma or respiration disease. The correct rates of the four knowledge points were different among different gender and the degree of education,which was higher in female than in male,and higher in high education degree than in the other(P

Nerve growth factor is one of the most important factors playing an important role in regulating the growth, development and survival of the neuron. The purified NGF from human placenta has been reported, the tissue from which can be isolated the NGF is very limited. It is important for basic research and clinic application to expression hNGF by genetic engineering. By polymerase chain reaction,gene fragment encoding the mature part of ?-NGF was amplified using the DNA of human placenta as template. The fragment was sequenced and inserted into expression vector pET-15b, and the recombinant expression vector pET15b-NGF was transformed into E.coli BL21(DE3)pLysS. After inducing with IPTG the NGF was higher expressed up to 25% of the total cell proteins. The expression product was purified with metal chelate affinity chromatography on Ni-IDA agarose under denaturing condition. The purity of rNGF was higher than 90% and yield of rNGF was 4.56mg/L expressing bacteria. SDS-PAGE revealed the NGF expression product had a Mr 16kDa. Western-blot displayed the recombinant product had strong immunological activity with rabbit anti-human ?-NGF polyclonic antibodies. The expression products were dealed with solubilizing inclusion bodies and refolding protein. The test of nerve fiber growth of chicken embryo DRG neurons displayed rNGF had biological activity.

Objective To determine the expression of membrane-bound B lymphocyte stimulator (BLyS) protein and its mRNA in vitro of peripheral blood mononuclear cells (PBMCs) from individuals with systemic lupus erythematosus (SLE),and to investigate the role of interferon-?(IFN-?) on the expression of BLyS.Methods PBMCs were obtained from 25 SLE patients (mean age of 31+14) and 20 healthy volunteers (mean age of 28?10).They were randomized into IFN-?(5 ng/ml) group and control group.PBMCs were col- lected at 0,6,12 and 24 h for BLyS mRNA assessment using semi-quantitative reverse transcription-PCR (RT-PCR).PBMCs were also collected at 72 h for membrane-bound BLyS protein detection using flow cy- tometry (FACS) and direct immunofluorescence.Results①The expression of BLyS mRNA and membrane- bound protein in PBMCs was significantly higher in individuals with SLE compared with healthy controls (P＜0.05);②IFN-?enhanced BLyS mRNA expression in PBMCs in both healthy controls and SLE patients,with the greatest effect at 6 h (stimulated vs unstimulated,0.42?0.19 vs 0.25?0.14,P＜0.01;0.59?0.28 vs 0.44?0.21,P＜0.01 );③IFN-?also increased the expression of membrane-bound BLyS protein in both healthy con- trols and individuals with SLE (FACs,mean fluorescence intensity,4.5+3.0 vs 3.7~2.6,P

Atrioventricular Block ; etiology ; Cardiac Catheterization ; adverse effects ; Heart Septal Defects, Ventricular ; therapy ; Hematologic Diseases ; etiology ; Hemolysis ; Humans

Objective To explore the characteristics of human plague using a sero-epidemiologic method in the source of the three rivers area in Qinghai for possible plague control strategies. Methods Investigate human plague sero-epidemiologically in the source of 4 counties in the three rivers area in Qinghai. The human serum would be tested to confirm the sew-positive rate for plague F1 antibody using indirect hemagglutination assay(IHA). Results A total of 2508 local participants were tested in 4 counties, the overall plague sero-positive rate was 2.31%(58/2508). This represents a statistically significant difference with 4 counties(X2=19.30,P<0.01). The sew-positive rate for males and females were 2.54% (32/1261) and 2.09% (26/1247), respectively. There were no statistically significant differences between males and females(X2= 0.65,P 0.05). The sero-positive rate in herdsman, cadre, Tibetan, Hart nationalities were 3.54% (44/1243), 6.47% (11 / 170), 2.40% (56/2335) and 1.47% (2/136), respectively. The sero- positive rate increased with age. The highest titre for human plague serum antibody was 1 : 640. Conclusion There were occult infections of plague in the population on source of three rivers area in Qinghai. Sero-epidemiologic data revealed that the human plague sero-positive rate was closely correlated with the local animal plague.

The aim of this study was to detect the nerve growth factor (NGF) level in serum and NGF low affinity acceptor CD271 expression on bone marrow leukemic cells in acute B lymphoid leukemia (B-ALL) patients and to analyze their clinical significance. The NGF level in serum and CD271 expression on leukemic cells in bone marrow were detected by enzyme linked immunosorbent assay and flow cytometry in B-ALL patients respectively. The results indicated that compared with control group, the NGF level in serum of patient group significantly increased (t = 4.191, p < 0.05), but CD271 expression on leukemic cells in bone marrow decreased significantly (t = 4.898, p < 0.05). The complete remission (CR) rate of 25 B-ALL patients was 64% (16/25) after one course of CVAD chemotherapy. There were statistically significant differences of NGF level and CD271 expression in non-remission (NR) group and control group (t = 3.976, p < 0.05 vs t = 5.052, p < 0.05), but there were no statistically difference of NGF level and CD271 expression in CR group (t = 1.102, p > 0.05 vs t = 1.150, p > 0.05) as compared with control group. The CD271 expression before and after chemotherapy between CR and NR groups showed statistically significant differences (t = 3.889, p < 0.05; t = 3.751, p < 0.05 and t = 4.678, p < 0.05 respectively), but NGF level before and after chemotherapy showed no statistical difference between these 2 groups (t = 0.476, p > 0.05). 50% (8/16) patients relapsed during following up, and of their NGF level [(168.00 ± 61.66) pg/ml] and CD271 expression [(52.29 ± 13.00)%] showed the significantly differences, compared with those in control group (t = 5.284, p < 0.05 vs. t = 6.073, p < 0.05), but the NGF level [(81.13 ± 25.32) pg/ml] and CD271 expression [(78.45 ± 7.12)%] of other 8 patients showed no statistical difference as compared with control group (t = 1.228, p > 0.05 vs t = 1.144, p > 0.05). Compared with low NGF level and CD271 low expression groups, the survival time of B-ALL patients with high NGF level and CD271 expression was not changed significantly (p = 0.750 vs p = 0.170). It is concluded that the increased NGF level in serum and decreased CD271 expression on bone marrow leukemic cells in B-ALL patients are related with leukemia development and may be the useful indexes to evaluate curative effect and prognosis.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Nerve Growth Factor ; blood ; Nerve Tissue Proteins ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; metabolism ; Receptors, Nerve Growth Factor ; metabolism ; Young Adult

OBJECTIVETo investigate the local cellular immune response after injection of superantigen, the highly agglutinative staphylococin (HAS), into the tumor bed after ultrasound-guided percutaneous microwave coagulation therapy (PMCT) in the liver cancer patients.

METHODSNinety-two patients with pathologically proven primary liver cancer were divided into two groups: 45 in group A were treated by PMCT alone and 47 in the group B by combined with ultrasound-guided percutaneous injection of highly agglutinative staphylococin (HAS). Before and after PMCT and HAS treatment, the patients underwent ultrasound-guided percutaneous biopsy from the tumor bed and the samples were examined by pathology and immunohistochemistry. The infiltration of CD3+, CD4+, CD57+ and CD68+ lymphocytes in treatment zone was compared between the two groups. Moreover, the infiltrating immunocytes were observed by transmission electron microscopy.

RESULTSOne week after HAS injection, the densities of CD3+, CD4+, CD57+ and CD68+ cells in the group B were 54.50 +/- 18.44, 38.14 +/- 12.44, 33.38 +/- 10.79 and 45.56 +/- 16.53, respectively. All the above mentioned parameters increased significantly in varying degrees compared with that before PMCT or HAS injection (P < 0.05). Four weeks after HAS injection, the density of CD3+, CD4+, CD57+ and CD68+ cells in the group B were 32.67 +/- 10.42, 23.43 +/- 6.99, 18.63 +/- 7.89 and 30.01 +/- 11.05, respectively, still significantly higher than those before PMCT (P < 0.05). Five weeks after PMCT and HAS injection, the densities of CD3+, CD4+, CD57+ and CD68+ cells in the group B were 54.50 +/- 18.44, 38.14 +/- 12.44, 33.38 +/- 10.79 and 45.56 +/- 16.53, versus 32.03 +/- 8.11, 15.67 +/- 8.32, 15.23 +/- 8.26 and 29.67 +/- 11.98 in the group A, respectively, still with a significant difference between the two groups (P < 0.05). A lot of lysosomes, endoplasmic reticulum and mitochondria in the immune cells after injection of HAS were observed by transmission electron microscopy.

CONCLUSIONThe local cellular immunity in liver cancer treatment area can be significantly improved by ultrasound-guided injection of highly agglutinative staphylococin after percutaneous microwave coagulation therapy.

Adult ; Aged ; Antigens, CD ; immunology ; Antigens, Differentiation, Myelomonocytic ; immunology ; CD3 Complex ; immunology ; CD4 Antigens ; immunology ; CD57 Antigens ; immunology ; Electrocoagulation ; methods ; Female ; Humans ; Liver Neoplasms ; pathology ; therapy ; Male ; Microwaves ; therapeutic use ; Middle Aged ; Superantigens ; therapeutic use ; T-Lymphocytes ; immunology

OBJECTIVETo study the effect of Tangshenkang Granule (TG) containing serum on renal mesangial cells' (RMCs) proliferation and TGF-β1/Smad2/3 pathway in the high glucose condition.

METHODSTwelve SD rats were randomly divided into four groups, i.e., the low dose TG group, the middle dose TG group, the high dose TG group, and the blank control group, 3 in each group. After 7-day gastrogavage via portal vein blood, rats were sacrificed and their serum samples were collected. RMCs were cultured in common rat serum and TG containing serum respectively. The proliferation of mesangial cells was determined by methly thiazolyl tetrazolium (MTT) assay to determine the optimal TG containing serum concentration. Expression levels of TGF-β1 mRNA and protein were determined by real time quantitative PCR and ELISA. Smad2/3 protein expression and phosphorylation were determined by Western blot and immunofluorescence.

RESULTSTG containing serum at different doses could inhibit high glucose induced RMC cells' proliferation, TGF-β1 over-expression and Smad2/3 phosphorylation.

CONCLUSIONTG containing serum could inhibit high glucose induced RMC cells' proliferation, and its mechanism might be possibly associated with inhibiting TGF-β1/Smad2/3 signaling pathway.

Animals ; Cell Proliferation ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; Mesangial Cells ; Phosphorylation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Serum ; Signal Transduction ; Smad2 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism