0.05),but the cross reaction rate of recombinant Ts21 protein with sera from mice infected with T2,T3,T4 and T7 was considerably lower than that of ES antigens(?2=17.069,P

Objective To observe the influence of electroacupuncure(EA) at different therapeutic time windows on the effects of inosine on neuronal apoptosis and expression of heat-shock-proteins-70 (HSP70)after focal cerebral ischemic reperfusion injury (CIRI) in rats.Methods The model was established by ligation of the artery for 2 hour.EA was delivered to “baihui” and “dazhui” through acupuncture needles 0.5 hr,2 hr,6 hr and 12 hr respectively following MCAO.The expression level of HSP70 and the number of apoptotic cells were examined by immunohistochemical technique and TUNEL,comparing the average optical density of HSP70 and the number of apoptotic positive cells of cortex penumbra and hippocampus CA1 area in rat brain.Results Compared with the model group,the average optical density of HSP70 in every EA group was increased in apoptotic positive cells of cortex penumbra and hippocampus CA1 area 0.5 h (89.98± 6.55),(128.73 ± 8.03),2 h (90.96±6.38),(132.25±8.78),6 h (93.71±6.12),(132.58±7.04),12 (96.19±7.30),(133.57±6.19)and the number of apoptotic positive cells of cortex penumbra in every EA group was decreased 0.5 h (1.80±0.84),2 h (3.40± 1.14),6 h (5.00± 1.00),12 h (5.00±2.45).The number of apoptotic cells of hippocampus CA1 area was decreased just in the EA group of 0.5 h (1.60± 1.89).Conclusion EA could increase the expression level of HSP70 and at the same time decrease the number of apoptotic cells,EA plays an important part in protecting the neuronal cells of brain after focal cerebral ischemic reperfusion injury.The acupuncture treatment should be taken as far as early.

Objective To express the antigen gene Ts21 of Trichinella spiralis, purify the recombinant protein and test its antigenicity. Methods T. spiralis gene Ts21 was sub-cloned into the prokaryotic expression vector pMAL-c2X and the recombinant pMAL-c2X-Ts21 was constructed. The recombinant plasmid was transformed into E. coli TB1 strain and induced by IPTG. The expression products were purified by MBP-binding affinity chromatography. The antigenicity of the recombinant protein was examined by SDS-PAGE and Western blotting. Mice were immunized with the recombinant protein, the titer of the immune sera was detected by ELISA. The distribution of Ts21 protein in muscle larvae was observed by IFA. Results The molecular weight of the expressed fusion protein was about Mr 63 500 and the expression level peaked at 4 h post-incubation. The portion of the fusion protein accounted for 18.2% of all the protein by thin-layer gel optical scanning. Western blotting demonstrated that the recombinant protein was recognized by sera from mice infected by T. spiralis (T1) and T. nelsoni (T7) as well as sera of patients with trichinellosis, but not by sera from mice in-fected with T. nativa (T2), T. britovi (T3) and T. pseudospiralis (T4). The recombinant protein did not react with sera from patients with ancylostomiasis, cysticercosis and schistosomiasis, but cross-reacted with sera from patients with parag-onimiasis, clonorchiasis and echinococcosis. High titers of antibodies were produced in mice immunized with the recom-binant protein. IFA showed that the Ts21 protein was mainly distributed in the cuticle of muscle larvae. Conclusion The Ts21 antigen gene of T. spiralis has been expressed and the recombinant protein shows antigenicity.

Aged ; Atropine ; Electrocardiography ; Female ; Heart Rate ; drug effects ; Humans ; Male ; Muscarinic Antagonists ; pharmacology ; therapeutic use ; Preoperative Care ; methods ; Quinuclidines ; pharmacology ; therapeutic use

Voltage-gated sodium channels (VGSCs) are known to be involved in the initiation and progression of many malignancies, and the different subtypes of VGSCs play important roles in the metastasis cascade of many tumors. This study investigated the functional expression of Nav1.5 and its effect on invasion behavior of human breast cancer cell line MDA-MB-231. The mRNA and protein expression of Nav1.5 was detected by real time PCR, Western Blot and immunofluorescence. The effects of Nav1.5 on cell proliferation, migration and invasion were respectively assessed by MTT and Transwell. The effects of Nav1.5 on the secretion of matrix metalloproteases (MMPs) by MDA-MB-231 were analyzed by RT-PCR. The over-expressed Nav1.5 was present on the membrane of MDA-MB-231 cells. The invasion ability in vitro and the MMP-9 mRNA expression were respectively decreased to (47.82+/-0.53)% and (43.97+/-0.64)% (P<0.05) respectively in MDA-MB-231 cells treated with VGSCs specific inhibitor tetrodotoxin (TTX) by blocking Nav1.5 activity. It was concluded that Nav1.5 functional expression potentiated the invasive behavior of human breast cancer cell line MDA-MB-231 by increasing the secretion of MMP-9.

Objective To optimize the experiment conditions of surface-enhanced Raman spectroscopy detection of serum fingerprint spectra.Methods Normal human serum was used as the sample and Ag nanoparticles as the active substrate.The enhanced signals of different optimized experiments were obtained , including serum dose(2.5 to 500 μl), incubation time(10 to 30 minutes) temperature(4℃,room temperature and 37℃),and different treatment(extraction and protein removal).Results and Conclusion Serum doses should not exceed 50μl.The ratio should range from 1∶1 to 5∶1, the incubation time is from 10 to 30 minutes, and the incubation temperature from 4℃ to 37℃.The signals of samples directly mixed with an active substrate are stronger than those of samples which are extracted or protein removed .

Adult ; Bone Neoplasms ; Humans ; Male ; Nose Neoplasms ; Osteoblastoma ; Skull Base Neoplasms

Recently surface-enhanced Raman spectroscopy (SERS) has been widely used in physics, chemistry and bio-medical science.Due to its high sensitivity and specificity,SERS is often used to detect changes in serum components in humans.Various biomolecules in human serum, such as proteins, lipids and nucleic acids, have their own distinctive raman spectroscopy so that different raman shift, band intensity and width reflect different metabolic abnormalities of cells at the molecular level in human serum.In this paper we described the general situation of SERS and summarized the latest research progress in a variety of diseases of human serum.Prospects of developmenls are also outlined.

Objective To study the variation characteristics of plasma von Willebrand factor antigen (vWF Ag) after the off-pump coronary artery bypass graft (OPCAB) and its predictive value on the assessment of cardiovascular ischemia events in the postoperative patients.Methods A total of 338 patients with non-ST-segment elevation myocardial infarction were selected from 2010 to 2012 in this retrospective cohort study,with 249 males and 89 females and a mean age of (69.2 ± 4.5) years.The level of vWF Ag was assayed by the IL ACL-TOP 700 blood coagulation instrument.Receiver-operator curve (ROC) analysis of vWF Ag levels in the prediction of risk of ischemic events was performed.x2 test and Logistic regression were conducted to analyze the relevance between vWF Ag and clinical pathological factors.Cox regression analysis model were used to evaluate the effect on prognosis.Results There was significant difference in vWF Ag level at different time point between the poor recovery group and the stable disease group (x2 =129.53 and 101.48 respectively,both P＜0.01).And the vWF Ag level was higher in the poor recovery group on the 14th,30th,60th,90th day after OPCAB than in the stable disease group at the same time points respectively (all P＜0.05).The optimum cut-off point of vWF Ag (the 30th day after OPCAB) for prediction of ischemic events was 251 ％ within the 31-90th day after OPCAB,the area under ROC curve was 0.839 (95％ confidence interval:0.776-0.902).The Logistic regression analysis showed that the vWF Ag level on 30th day after OPCAB was affected by age,left ventricular ejection fraction,left main artery disease,serum creatinine level,vascular number of bypass grafts,history of myocardial infarction and hypertension (all P＜0.01).The Cox analysis showed that the cut-off value of vWF Ag (on the 30th day after OPCAB) was the independent prognostic assessment parameter for ischemic events in patients with cardiovascular diseases within one year.Conclusions The variation of serum vWF Ag level can reflect the integrated influence of multiple pathological factors on vascular endothelial function after the OPCAB,it can become the effective predictor for disease progression within 90 days and the long-term prognosis after OPCAB.

Objective To explore the association between acute ischemic stroke and albumin to creatinine ratio (ACR).Methods A case-control study.During January to December in 2013, 127 acute ischemic stroke patients as case group and 150 controls who were similar with case group in age and gender were recruited in Tianjin Union Medicine Center . According to diabetes , hypertension , cardiovascular diseases and patients without these three diseases , case group was divided into A1, B1, C1 and D1 subgroups, control group was divided into A2, B2, C2 and D2 subgroups in the same way.The first morning urine from participants were collected .Urinary albumin concentration was tested by nephelometry , urinary creatinine was examined by using enzymic method , ACR were calculated(≥30 mg/g as the cutoff value). Then difference of ACR between case and control group was compared , the cutoff value of albuminuria for ischemic stroke patients was analyzed by ROC , and the risk factor of ischemic stroke were analyzed by logistic regression analysis.Results The prevalence of albuminuria in ischemic stroke patients was 38.58%(49).According to analysis of covariance, after adjustment for age, gender, cardiac-disease, diabetes, hypertension, lnACR in case group was significantly higher than control group (3.18 mg/g vs 2.78 mg/g, t=2.13 P=0.03), especially D1 was significantly higher than D2 subgroup (3.01 mg/g vs 2.51 mg/g,t=5.56,P=0.009) .If 19.82 mg/g from ROC analysis was used as the cut-off value, the sensitivity and specificity are 68.5% and 61.7%.The odds ratio ( OR ) of ACR >19.82 mg/g was about 2-fold when compared with ACR<19.82 mg/g adjusted for stroke risk factors , and the OR value is 2.43 in comparison of patients without diabetes , hypertension and cardiovascular diseases .Conclusions Urinary albumin excretion is the independent risk factor of ischemic stroke .The increased urinary albumin has important clinical significance to predict the risk of ischemic stroke for the patients without diabetes , hypertension and cardiovascular diseases.