Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Spinocerebellar Ataxias ; therapy ; Treatment Outcome ; Young Adult

Objective:To construct an adenovirus-mediated anti-sense RNA targeting K-ras exon 1 of SW1990 cell line and observe its effect on ceil proliferation and apoptosis after transferred into SW1990 cell line.Methods:K-ras exon 1 cDNA was cloned into shuttle vector pShuttle-CMV and the resultant plasmid was confirmed by enzyme digestion and PCR.Clones with inverted insertion were selected and co-transferred into E.coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1 to produce recombinant plasmid by homologous recombination.Recombinants were then selected and transfected into 293 cell line to produce recombinant adenovirus.Recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified;the virus titer was determined.Ad-LacZ was used to infect SW1990 cells and the infection efficiency was observed by X-gal staining.SW1990 cells was infected with the recombinant adenovirus and their proliferation and apoptosis were determined by MTT and annexin V/PI FCM assay.Results:A 282 bp target gene fragment was acquired by PCR;the titer of recombinant adenovirus was 7.6?10~8 pfu/ml before purification by CsCl_2 gradient centrifugation and 5.0?10~(10)pfu/ml after CsCl_2 gradient centrifugation.When the recombinant adenovirus was at 100 MOI,the infection efficiency of SW1990 cells nearly reached 100%.The transfection of recombinant adenovirus significantly inhibited SW1990 cell proliferation(P

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal disorder of hematopoiesis due to the inactivation of PIG-A gene. However, the presence of mutant PIG-A gene in a group of hematopoietic cells is not enough for the development of PNH, immunologic injury and apoptotic effects are considered to play an important role in clonal expansion. Knowledge of the molecular mechanisms leading to PNH has substantially increased in the past decades, which remarkably advances the diagnostic modalities and treatment approaches of patients with PNH. Though great progress has been made because of targeted therapy method, the challenges are still ahead. In this review the advances of studies on mechanism, laboratorial diagnosis and therapeutic protocols of PNH are summarized.
Complement System Proteins ; Hemoglobinuria, Paroxysmal ; diagnosis ; genetics ; pathology ; therapy ; Humans ; Membrane Proteins ; genetics ; T-Lymphocytes, Regulatory

Objective To compare the effects of clopidogrel with or without combined with CYP3A4-metabolized statin in treating coronary artery disease (CAD) among the elderly patients.Methods The study cohort was defined as all patients were over 60 years of age and hospitalized for CAD who were prescribed clopidogrel between January 2000 and February 201 1.A total of 1021 patients were enrolled,with 178 of them prescribed clopidogrel and 843 patients were administrated clopidogrel combined with statins (CYP3A4-metabolized statins 636 and non CYP3A4-metabolized statins 207).The primary endpoint was all cause of death and the second endpoint were non-fatal myocardial infarction (MI),but hospitalized for unstable angina,stroke,transient ischemic attack,or repeated revascularization (PCI or coronary artery bypass graft).Results Among the clopidogrel group and the clopidogrel plus statins group,the incidence density of death was 6.86/1000 and 3.18/ 1000 respectively,with crude RR as 2.15(95％CI:1.39-3.33) and statistically significant (x2=3.53,P＜0.01).The incidence density of composite thromboembolic events did not show statistical significance (P＞0.05).The two groups were 1∶1 matched,after propensity score matching,clopidogrel coadministrated with statins group showed significant decrease in all cause of death,with RR as 0.42 (95％ CI:0.19-0.93),x2=7.23,P＜0.01.No significant difference was observed in deaths or composite thromboembolic events between statins via different cytochrome P450 pathways.Conclusion Clopidogrel with statin could reduce the mortality of elderly CAD patients compared with clopidogrcl without statin.The result did not show statistical significance between CYP3A4-metabolized statins or non CYP3A4-metabolized statins regarding the mortality or composite endpoint events.

Objective To investigate nursing students' knowledge of the different stages of practice, nursing education and hospital occupational protection against blood-transmitted diseases, for the future of nursing students in vocational prevention education basis.Methods Survey methods were used in 2009 to 2010 in 15 upper-second in Yancheng city general hospital, 344 nursing students of occupational protection awareness was investigated.Results Nursing students were different in occupational protection. Undergraduate nursing students in career awareness and prevention are better than college, secondary school nursing students(F=6.74,54.37;P<0.01). Professional knowledge and awareness level of protection of nursing students in three grade hospitals and two grade hospitals was significantly different(t=2.46,8.85;P<0.05 of P<0.01).Conclusions The higher the level of education and hospitals, the longer the training, nursing students was stronger awareness of occupational protection. Occupational protection of nursing students were generally poor execution, it was needed to strengthen education and training of schools and teaching hospitals. During practice nursing students should be strengthened the execution of occupational protection.

Objective：This study was designed to evaluate the application of three markers in diagnosing lung cancer and calculate Bergman Parameter to raise the sensitivity and validity in detecting the lung cancer. Methods：Using immunoradiometric and chemiluminescence assay, we observed serum NSE (neuron-specific enolase), CEA (carcinoembryonic antigen), and CA50(cancer-associated carbohydrate antigens 50) in 107 cases of lung cancer and 118 cases of benign pulmonary diseases. Bergman parameter was calculated with Bergman’s formula. Results：The sensitivity of NSE, CEA, CA50 for lung cancer were 51.4％ , 54.2％ , and 31.7％ , respectively. The specificity was 85.7％ , 89.8％ , and 93.0％ , respectively, while diagnostic accuracy rates were 64.3％ , 72.9％ , and 55.8％ . The sensitivity, specificity and diagnostic accuracy rate of Bergman parameter (95.2％ , 87.5％ and 92.1％ ) were higher than these three markers in lung cancer (P<0.05). We found that Bergman parameter could be used to moniter the prognosis of advanced lung cancer. Conclusion： Serum NSE, CEA, and CA50 are of value in detecting lung cancer. Bergman parameter, calculated with NSE, CEA, CA50, is able to increase specificity and accuracy in diagnosis of lung cancer.

OBJECTIVETo investigate the biological and clinical features of Chinese rhesus monkeys after intravenous (IV) and intrarectal (IR) challenge with SIVmac239 in rhesus monkeys of Chinese origin, and compare the differences between the routes of infection.

METHODSRhesus monkeys of Chinese origin were inoculated with SIVmac239 either by IV (n = 19) or IR (n = 6) routes. Simian immunodeficiency virus (SIV)-specific antibody titer, CD4 + T cell counting, plasma SIV load, lymph node pathology, and clinical manifestations were compared between these two groups 232 or 168 days after challenging.

RESULTSAll SIVmac239-inoculated animals became seropositive for SIV-specific antibodies. SIV-specific IgM was detected in IV groups as from day 10 but was not detected in IR for all the time points. Although SIV-specific IgG was detected as from day 30 in both groups, the IgG titers were ten-fold higher in IV group than in IR group after day 168. CD4 + T-cell counting decreased progressively in IV group but remained stable in IR group over time. Plasma SIV RNA loads peaked in all animals between day 10 and day 14 (10(7) copies/ml), then declined to "setpoint" (10(3) - 10(6) copies/ml) about 2 months later. Most inoculated animals manifested lymphadenopathy. Two animals in IV group and one in IR group died of simian AIDS between day 150 and day 210, as evidenced by the autopsies showing the depletion of lymph tissues, Pneumocystis carinii pneumonia and other opportunity infections. Conclusion IV or IR inoculation of SIVmac239 in Chinese rhesus monkeys will result in chronic SIV infection with a similar clinical feature of natural HIV infection, which provides an excellent experimental animal model for AIDS.

Animals ; CD4-Positive T-Lymphocytes ; metabolism ; China ; Disease Models, Animal ; Female ; Macaca mulatta ; virology ; Male ; Rectum ; virology ; Simian Acquired Immunodeficiency Syndrome ; immunology ; virology ; Simian Immunodeficiency Virus ; immunology ; pathogenicity ; Veins ; virology

OBJECTIVE Evidience appears that parthenolide (PN) induces anti-tumor effects by NF-κB signal pathway. MCL3 the derivative of PN,is sesquiterpene lactone synthesized by the group of Professor Pan Xiandao.The study was to explore the anti-tumor activity and mechanism of MCL3 in glioma. METHODS The effect of MCL3 on the proliferation of glioma cell lines was examined by MTT assay. Apoptotic activity was investigated by flow cytometry. The Transwell cell invasion assay was used to determine the effect of MCL3 on the G422 cell invasive ability.The effect of MCL3 on the angio-genesis was analyzed by a capillary-like tube formation assay. The subcutaneously transplanted and orthotopic G422 cell xenograft models were used to detect the effect of MCL3 on tumor growth in vivo. The pathological changes were analyzed by H&E staining. Protein level related to the NF-κB signal pathway was dertimined by Western blotting. The effect of MCL3 on the NF-κB transcriptional activity was examined by a dual-luciferase reporter assay.RESULTS The anti-proliferative activity was observed following treatment with MCL3 for 96 h in G422, U-87 MG, U251 and Hs683 cell lines, and the IC50 was 8.94 μmol·L-1,6.44 μmol·L-1,14.8 μmol·L-1,18.9 μmol·L-1,respectively.The percentage of apop-totic cells increased in MCL3-treated G422 cells,and the apoptosis rate was 26.4%(the apoptosis rate was 5.68% in control group).MCL3 could inhibit the invasion in G422 cells,and the invasive inhibition rate was 43.63%(P<0.01)at 10.0 μmol·L-1.MCL3 inhibited tube formation of EA.hy926 cells,and the inhibitory rate was 81.67%(P<0.01)at 10.0 μmol·kg-1.At 40.00 mg·kg-1,MCL3 supressed tumor growth by 79.03% (P<0.01) in tumor weight in subcutaneously transplanted G422 xenograft models, and by 69.97% (P<0.01) in volume in orthopotic G422 xenograft models. H&E staining demonstrated that MCL3 could decrease tumor angiogenesis and invasion, increased necrosis of tumor cells. The dual-luciferase reporter assay showed that MCL3 inhibited NF-κB transcriptional actvity, and the inhibition rate was 50.07%(P<0.05)at 10.0 μmol·L-1compared with control.Moreover,MCL3 inhibited the phos-phorylation of NF-κB in nuclear mediated by supression of phosphorylated IKKα/β and IκB,and decreased the expression of IL-6 regulated by NF-κB.Eventually,the phosphorylation of State3 decreased following the administration of MCL3, resulting in the downregulation of State3 taget genes, including HIF, VEGF,FAK,MMP-2,MMP-9,Bcl-2 and Bcl-xL.CONCLUSION The anti-tumor effect of MCL3 was partly due to the inhibition of NF-κB/IL-6/State3 pathway in glioma.

AIMTo evaluate the roles of puerarin in alleviating the STZ-induced lung injury.

METHODSDM model was established by streptozotocin (STZ) intraperitoneal injection to study the injury mechanisms of the lung. SD rats were divided randomly into control group (C group), diabetes group (DM group), diabetes + puerarin group (DM + Pur group). The blood glucose and weight were observed and recorded before and the 20 th, 40 th, 60 thd after administration of saline, STS, STZ+ Pur. Contents of NO and malondialdehyde (MDA) and activity of superoxide dismutase (SOD) were measured in lung tissues. Light microscope (LM), transmission electron microscope (TEM) and immunohistochemical analysis were also used.

RESULTS(1) Compared with control group, the contents of NO and MDA were increased significantly (P < 0.01), while the activity of SOD reduced (P < 0.05). Compared with DM group, treatment with puerarin inhibited the increase of NO level (P < 0.01), and MDA content began to decline from 40 days after the model was established (P < 0.01), and inhibited the decrease of SOD activity induced by DM (P < 0.01). (2) LM and TEM results showed that alveolar and capillary basement membrane became thick, the number of tiny villus decreased markedly, the quantity of osmiophilic multilamellar body reduced remarkably, hyperplasia was shown in collgen fibre. Puerarin could alleviate above injuries induced by DM. (3) Immunohistochemical staining results showed that mild brown positive stain of NT could be seen in protoplasm of lung tissues. STZ administration induced the expression of NT in the protoplasm of cells, and led to stronger positive signals of NT than that of control group. Treatment with puerarin weakened the positive stain of NT.

CONCLUSION(1) DM induced by STZ leads to a significant and sustained increase in blood glucose and obvious lung injury, which may be associated with the overproduction of free radicals. (2) The pathway of NO/ONOO- is one of the injury mechanisms of the lung tissues cells. (3) Puerarin suppresses the expression of NT and elevates the activity of SOD. Thereby, resulting in the reduces of the production of free radicals, which may be one of the mechanisms of its anti-oxidative-injuries.

Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Isoflavones ; pharmacology ; Lung ; metabolism ; pathology ; Lung Injury ; chemically induced ; Malondialdehyde ; analysis ; Nitric Oxide ; analysis ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

The aim of the present study is to explore the mechanism of estrogen on regulating cardiac function disorder by adjusting the stimulating adenylate cyclase G α protein (Gαs)-cycle adenosine monophosphate (cAMP) signal pathway. Adult female rats were randomly divided into five groups: sham group, ovariectomized group (OVX), OVX and 17β-estradiol given group (OVX+E₂), OVX and isoprenaline injected group (OVX+ISO), OVX and 17β-estradiol, isoprenaline injected group (OVX+E₂+ISO). Rats were ovariectomized, and two weeks later, OVX+E₂group was injected with E₂, OVX+ISO group was injected with ISO, OVX+E₂+ISO group was injected with E₂and ISO. Another four weeks later, the hemodynamic parameters were monitored by carotid artery intubation: left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), maximal differentials of left ventricular developed pressure (+dp/dt(max)), and minimal differentials of left ventricular developed pressure (-dp/dt(max)). Brain natriuretic peptide (BNP) and cAMP concentration in plasma were determined; Gα(s) protein expression in myocardium was determined. The results showed that the hemodynamic parameters, the concentration of BNP and cAMP in plasma had no significant changes after ovariectomy compared with sham group. But after isoprenaline injection in ovariectomized rats, LVSP and +dp/dt(max) declined (P < 0.01), LVEDP and -dp/dt(max) elevated (P < 0.01); plasma BNP concentration increased (P < 0.01); plasma cAMP concentration decreased (P < 0.01), compared with OVX group. Further estrogen supplements improved the heart function treated by isoprenaline: LVSP and +dp/dt(max) elevated (P < 0.01), LVEDP and -dp/dtmax declined (P < 0.05, P < 0.01); the plasma BNP concentration decreased (P < 0.01); the plasma cAMP concentration increased (P < 0.01). Estrogen had no significant influence on Gαs protein expression. The results suggest that estrogen can alleviate myocardial injury and regulate cardiac function disorder by increasing cAMP level, finally improved the excessive suppression of myocardium.
Animals ; Cyclic AMP ; blood ; Estradiol ; pharmacology ; Estrogens ; pharmacology ; Female ; GTP-Binding Protein alpha Subunits, Gs ; metabolism ; Hemodynamics ; Isoproterenol ; adverse effects ; Myocardium ; pathology ; Natriuretic Peptide, Brain ; blood ; Ovariectomy ; Rats ; Signal Transduction