Objective Serologic proteome analysis method (SERPA) was used to compare the different of the serum proteins between normal and pancreatic cancer patients' serum, and to find a new specific pancreatic cancer marker. Methods HPLC was used to eliminate albumin from the serum, two-dimensional electrophoresis was used to separate the proteins. After imaging collection and analysis, the different proteins between pancreatic cancer and normal subjects were cut for mass spectrometry. Results Four discrepancy proteins were obtained. Guanylate cyclase-activating protein 2 was highly expressed in normal serum but not pancreatic cancer. Hp2-alpha, transthyretin and KIAA1018 protein were highly expressed in cancer patients'serum but not normal people. Conclusions KIAA1018 may become a promising tumor marker for screening and early diagnosis of pancreatic cancer.

Objective To investigate the possible mechanism of the gap junctional influence on the change in permeability of the blood-brain barrier(BBB)after reperfusion subsequent to cerebral ischemia.Methods In the test laser scanning confocal microscope(LSCM)was used to investigate the change of Cx43 levels and distribution.The MCAO/R model was induced using intraluminal suture technique first described by Longa with a little modification.A total of 60 Wistar rats were divided into 4 groups:the sham-operation group,control group,octanol-treatment group and DMSO vehicle control group. Control group were further divided into seven subgroups at different time points of reperfusion after middle cerebral artery occlusion.To observe the change in permeability of BBB,Evans blue(EB)in the brain tissue was surveyed by the means of EB fluorescent quantitation.Octanol-treatment group and DMSO vehicle control group were done at the point of the peak of permeability of BBB.Octanol,the specific blocker for gap junctions(GJ)was used in an intervention study.To compare the amount of EB with the same point of groups,the influence of octanol on BBB permeability was investigated.Results At 3 h of reperfusion after cerebral ischemia for 2 h,the permeability of BBB began to increase,reached the peak at 24 h of reperfusion and was still elevated at 72 h.The Cx43 expression formed into bigger plague and remained linear disposition in the penumbra after reperfusion subsequent to cerebral ischemia.Octanol group was done at 24 h of reperfusion after cerebral ischemia.The amount of EB of octanol group((4.924?0.296)?g/g)was significantly lower than that of corresponding operation control group(5.543?0.506)?g/g.Conclusions (1)Cx43 expression is concentrated around vessels in brain.The Cx43 forms into bigger plague and the function maybe strengthens after reperfusion.Gap junction might aggravate the disruption of BBB.(2) Octanol,the specific blocker of gap junctions,could effectively prevent the permeability of BBB from increasing and has a protective effect on BBB.

Objective To explore the value of echo-tracking technique in the detecting the ipsilateral external iliac artery elasticity after gunshot wound in pig limbs.Methods Two-dimensional ultrasound imagings and elasticity parameters of external iliac artery of fifteen pig limbs were obtained respectively,and the results were compared with pre-injury groups.The animals were sacrificed after ultrasonography.Pathological examinations of adjacent external iliac artery of injured pig limbs were analyzed.Results Two-dimensional ultrasound imagings of external iliac arteries had no significant changes post-injury.The changes of elasticity parameters were significiant differences in the injured group comparing with the pre-injured group (P ＜0.05),including the increased stiffness parameter(β),pressure-strain elasticity modulus (Ep)and the decreased arterial compliance(AC).Pathological result showed that the internal elastic lamina of artery detected were flat,endothelial cells came off discontinuously and structure of them were undefined.Conclusions Echo-tracking technique can find the elastic changes of adjacent artery indirect injured by gunshot wound sensitively and which can suggest the occurrence of vascular indirect injury.

Adult ; Aged ; Angina Pectoris ; blood ; drug therapy ; Angiotensin II ; blood ; Capsules ; Diagnosis, Differential ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Endothelins ; blood ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy

OBJECTIVETo prepare anti-recombinant protein antibody from immunized mice with recombinant nucleocapsid protein (NP) of human influenza A3 (IFV-A3) virus expressed in prokaryotic cell, and to explore the feasibility of utilizing anti-recombinant protein antibody to detect influenza A virus.

METHODSNP genes of human influenza A virus were analyzed with computer softwares of ClustalX, Antheprot, et al. to determine the antigenicity in conserved regions. Three different partial NP genes were harvested and cloned into pET-28(c) plasmid, the recombinant plasmids were induced to express partial NP segments in BL21 cells. The recombinant proteins were purified with Ni-agarose by affinity chromatography and immunized BALB/c mice. The polyclonal antisera harvested from mice were analyzed with Western Blot and immunohistochemistry assays to detect the reactions with IFV-A.

RESULTSThree recombinant plasmids were expressed with high yield in BL21 cells, about 15-20 mg/L. Western Blot results indicated that the three prepared antisera (1:2000) positively reacted with NP from IFV-A3-infected cells. And immunohistochemistry assays suggested that anti-NP1, anti-NP2, anti-NP3 antisera positively reacted with IFV-A3 or IFV-A1-infected MDCK cells, with titers of 1:640 to 1:1280.

CONCLUSIONThe recombinant NP of IFV-A3 would induce polyclonal antibodies with high titers in mice. The polyclonal antibodies would cross-react with IFV-A3 and IFV-A1. It is feasible to predict the antigenicity with systematical bioinformatics analyses and then induce anti-IFV antibodies with high dilutions, and it is possible to be utilized in the early detection and subtyping analyses of IFV-infections.

Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Influenza A virus ; genetics ; immunology ; Influenza, Human ; immunology ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology

OBJECTIVETo compare the immediate effect of moxibustion on Yanglingquan (GB 34) on body surface thermograph between the patients with myofascial pain syndrome (MPS) and healthy people.

METHODSThirty-five cases of MPS were selected in observation group and 15 cases of healthy people were in control group. ATIR-M301 medical infrared imaging device was used to observe the changes in body surface thermograph before and 15 min after moxibustion on Yanglingquan (GB 34) The qualitative analysis of picture pattern and the quantitative analysis of temperature were integrated to compare the differences between two groups.

RESULTSConcerning to the infrared thermograph expression on the back of MPS cases in observation group, the abnormal thermal zone presented in 88.6% of the cases, mostly distributed in the upper and secondly in the lower back. The spinal thermal line was broken or disappeared ine about 50% of cases. The location of pain complained by 71.4% of cases was coincident with the zone of infrared thermograph expression. There was statistical significant difference in the temperatures difference of the two sides in the upper abnormal thermal zone on the back in observation group as compared with control group (P < 0.001). Body surface thermograph on the back were impacted by moxibustion on Yanglingquan (GB 34) for either MPS patients or healthy people, especially in upper back and rhomboid fossa. Moxibustion on Yanglingquan (GB 34) increased much more apparently the temperature on the upper back in MPS patients as compared with healthy people (P < 0.05).

CONCLUSIONMoxibustion on Yanglingquan (GB34) brings the immediate improvements in the back thermograph for MPS patients.

Acupuncture Points ; Adult ; Aged ; Body Temperature ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Myofascial Pain Syndromes ; physiopathology ; therapy ; Thermography ; Young Adult

OBJECTIVETo investigate the effects of hydroxycamptothecin (HCPT) on proliferation and apoptosis of rat hepatic stellate cells (HSC).

METHODSRat HSC line (HSC-T6) and rat hepatocyte line (BRL-3A) were treated with different concentrations of HCPT (0, 0.008, 0.016, 0.031, 0.125, 0.25, 0.5, 1, 2, 4, 8, 16, 32 mg/L respectively) for 24 h. Cell proliferation was assessed by MTT colorimetric assay, apoptosis was detected with PI staging followed by flow cytometry, and by DNA ladder assay. The morphological change of apoptosis was observed under transmission electron microscopy (TEM).

RESULTSMTT assay indicated that HCPT significantly inhibited the proliferation of HSC-T6 and BRL-3A in a dose-dependent manner. 24 h after the treatment with different concentrations of HCPT (0.25, 0.5, 1 mg/L), the apoptosis rate (13.46%+/-2.42%, 26.25%+/-5.65%, 47.05%+/-8.76%, respectively) in HSC-T6 was significantly higher than that in control cells (4.89%+/-1.80%, F = 34.24, P less than 0.01). 24 h after 0.5 mg/L HCPT treatment, cell shrinkage, nucleoli disappearance, chromatin condensation were found under TEM, and DNA ladder was demonstrated by agarose gel electrophoresis.

CONCLUSIONHCPT could significantly inhibit proliferation and induce apoptosis of HSC-T6 in a dose-dependent manner.

Animals ; Apoptosis ; drug effects ; Camptothecin ; analogs & derivatives ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Hepatic Stellate Cells ; drug effects ; Rats

OBJECTIVETo observe the effect of beta(3)-adrenoceptor (AR) in regulating resting intracellular Ca(2+) concentration of the ventricular myocytes and investigate the signaling pathway in rats with experimental heart failure.

METHODSRat models of experimental heart failure were established by ligation of the anterior descending artery, and the myocytes were isolated by enzymatic digestion. The resting intracellular Ca(2+) concentration was determined using laser scanning confocal microscopy (LSCM) in the cells stimulated with 1 micromol/L BRL37344 (a selective beta(3)-AR agonist) alone or in combination with PTX, L-NAME, or methylene blue.

RESULTSIn the ventricular myocytes from normal control rats, BRL373444 reduced the resting intracellular Ca(2+) concentration of by 45.5%, while the reduction increased to 59.4% in the cells from rats with heart failure. In combination with L-NAME (10 micromol/L), methylene blue (10 micromol/L), and PTX (2 microg/ml), BRL373444 caused a reduction in resting intracellular Ca(2+) concentration of the ventricle myocytes from normal control rats by 10.1%, 16.9%, and 15.4%, respectively in control group, while the rate was 16.9%, 19.3%, and 11.7% in the heart failure group.

CONCLUSIONSBeta(3)-AR agonist can decrease the resting intracellular Ca(2+) concentration of the ventricular myocytes, but the reduction is smaller in cells from rats with heart failure than in cells of normal rats. This effect is mediated through the PTX-NOS-NO pathway.

Adrenergic Agonists ; pharmacology ; Adrenergic beta-3 Receptor Agonists ; Animals ; Calcium ; metabolism ; Heart Failure ; chemically induced ; metabolism ; pathology ; Heart Ventricles ; pathology ; In Vitro Techniques ; Intracellular Space ; drug effects ; metabolism ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Rats ; Rats, Wistar ; Receptors, Adrenergic, beta-3 ; metabolism ; Rest ; Signal Transduction ; drug effects

BACKGROUND AND OBJECTIVECombined hypoxic cytotoxic drugs and chemoradiotherapy is an important mean of oncotherapy, and Tirapazamine (TPZ) is one of the most remarkable drugs. It has been shown that TPZ has a synergistic effect with radiotherapy on tumor cells, but whether TPZ would down-regulate the expression of the hypoxia-induced genes has not been reported. This study was to investigate the hypoxia-induced mRNA expressions of hypoxia inducible factor-1alpha (HIF-1alpha) and osteopontin (OPN) in human nasopharyngeal carcinoma HNE-1 and CNE-1 cells and the radiosensitization of TPZ, a hypoxia-specific drug, on HNE-1 and CNE-1 cells in vitro.

METHODSThe IC50 values of TPZ for HNE-1 and CNE-1 cells were measured using MTT assay, and the mRNA expressions of HIF-1alpha and OPN in HNE-1 and CNE-1 cells was determined using RT-PCR under aerobic and hypoxic conditions, respectively. The survival rates of HNE-1 and CNE-1 cells treated with or without TPZ at IC10 in the presence or absence of oxygen for 6 h were determined using colony formation assay following exposure to 1-6 Gy of 60Co radiation. The dose-survival curves were plotted and the values of D0, Dq and SER were calculated as a single-hit multitarget model.

RESULTSThe IC50 values of TPZ were 34.81 μmol/L and 35.02 μmol/L in HNE-1 and CNE-1 cells under aerobic condition, and 30.20 μmol/L and 28.48 μmol/L under hypoxic condition, respectively. The expressions of HIF-1alpha and OPN mRNA were reduced by TPZ in HNE-1 cells, but not in CNE-1 cells under hypoxic condition. For the HNE-1 cells, the respective values of D0 and Dq were 0.89 Gy and 0.28 Gy following normoxic irradiation versus 1.47 Gy and 0.44 Gy following hypoxic irradiation. For the CNE-1 cells, the respective values of D0 and Dq were 0.72 Gy and 0.68 Gy following normoxic irradiation versus 0.95 Gy and 0.56 Gy following hypoxic irradiation. The values of D0 and Dq for HNE-1 and CNE-1 cells treated with TPZ under hypoxic condition following irradiation were 0.66 Gy, 0.21 Gy and 0.85 Gy, 0.79 Gy, respectively.

CONCLUSIONTPZ can down-regulate hypoxia-induced expression of HIF-1alpha and OPN mRNA of HNE-1 cells and radiosensitize the HNE-1 cells but not CNE-1 cells, and act as a hypoxia modifier.

Antineoplastic Agents ; pharmacology ; Cell Hypoxia ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Cobalt Radioisotopes ; Down-Regulation ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Inhibitory Concentration 50 ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Osteopontin ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Radiation Tolerance ; drug effects ; Radiation-Sensitizing Agents ; pharmacology ; Triazines ; pharmacology

OBJECTIVETo explore the mechanism of Shenqi compound recipe (SQCR) anti-earlier diabetic artherosclerosis in GK rats.

METHODFour-month specefic pathogen free (SPF) GK rats were divided randomly according to blood glucose level into four groups: model group (5 mL x kg(-1) x d(-1) sterile water), ramipril group (positive control, 1 mg x kg(-1) x d(-1)), SQCR low dosage (0.72 g x kg(-1) x d(-1)) and SQCR high dosage group (2.88 g x kg(-1) x d(-1)) and Wistar rats as normal control group(5 mL x kg(-1) x d(-1) sterile water). GK rats took high-fat diet freely and meanwile were injected N-omega-nitro-L-arginine methyl ester (L-N-AME) intra-peritoneally with the dose of 10 mg x kg(-1) x d(-1) in order to induce earlier diabetic artherosclerosis, while normal control group took regular diet and were injected normal saline intra-peritoneally. In the experiment periods, each group was administrated correspondent substance respectively for 32 d. At the end, sampling blood by abdominal aorta and picking aorta on ice. Determined monocyte chemoattractant protein-1 (MCP-1) concentration by ELISA, messenger ribonucleic acid (mRNA) expression of MCP-1 and peroxisome proliferator-activated receptor gamma (PPARgamma) in aorta by reverse transcriptase PCR (RT-PCR).

RESULTConcentrations of MCP-1 in serum in SQCR low and high dosage groups and the mRNA expression of MCP-1 in SQCR high dosage group were all decreased significantly compared with model group (P < 0.05). The mRNA expression of PPARgamma in SQCR low and high dosage groups all increased compared with model group (P < 0.05 or P < 0.01).

CONCLUSIONInhibiting the mRNA and protein expression of MCP-1 and upregulating the mRNA expression of PPARgamma in aorta might be contribute to SQCR anti-earlier diabetic artherosclerosis in GK rats partly.

Animals ; Aorta ; metabolism ; Astragalus membranaceus ; chemistry ; Atherosclerosis ; etiology ; metabolism ; Chemokine CCL2 ; biosynthesis ; blood ; genetics ; Diabetes Mellitus, Type 2 ; complications ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Male ; PPAR gamma ; biosynthesis ; genetics ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar