Streptococcus gordonii is a nonpathogenic gram-positive commensal bacterium and component of the normal microbial flora of the human oral cavity. It is suitable to be as a mucosa vaccine vector due to its special bionomics. knowing the bionomics of Streptococcus gordonii,the general expressing system,and the application of it in mucosa vaccine,will provid important reference for the further development of its mucosa vaccine.

To construct expressing vector carrying esat6 gene and express this protein in Streptococcus gor-donii GP251. esat6 gene was amplified by PCR with specific primer from genome of Mycobacterium tuber-culosis (MTB)H37Rv. Inserted esat6 into the pMD18-T vector by T/A clone to get recombinant vector pMD18-esat6. Then digested pMD18-esat6 with restriction enzyme, esat6 was cloned to vector PSMB104 and expressed in Streptococcus gordonii GP251. The expression of esat6 protein was detected by Tricine-SDS-PAGE and Western-blot, ELISA technique was also used to detect its secretory volume. Re-striction endonuclease, PCR, Tric ine-SDS-PAGE and Western-blot confirmed that esat6 gene was cloned into expressing vector successfully, and a 10 kD protein secreted in Streptococcus gordonii GP251, this pro-tein has a good immunogenicity. The expression vector of esat6 gene was constructed, and esat6 protein ex-pressed in Streptococcus gordonii1 successfully, it will be benefit for future study.

BACKGROUND: Direct observation of infarcted microvaseular perfusion after cerebral infarction is the most immediate evidence in evaluation of therapeutic effects.OBJECTIVE: To observe dynamic changes in flowing quality and field in cerebral ischemic area in rats so as to qualitatively and quantitatively analyze the microvascular state in pia matral encephali ischemic area and the changes of associated leukocytic flowing velocity and flowing morphology.DESIGN: Randomized controlled experiment was designed.SETTING: Department of Acupuncture and Moxibustion of General Hospital Chinese PLA.MATERIALS: The experiment was performed in Research Room of Microcirculation of General Hospital Chinese PLA, in which, 120 female Wis tar rats of 2-month old were employed and randomized into acupuncture group (36 rats), model group (36 rats), sham-operation group (36 rats) and normal group (12 rats). The first 3 groups were subdivided in 3 hours, 6hours and 24 hours groups according to the prescriptive time divisions, 12rats in each group.METHODS: In acupuncture and model groups, after abdominal anesthesia with pentobarbital sodium, the cranium was opened and the occlusion of meddle cerebral artery was performed in cerebrum with heat-coagulation method. In sham-operation group, anesthesia and cranium opening were performed, but the meddle cerebral artery was not occluded. In normal group, no any management was performed. In acupuncture group, 60 minutes after occlusion of meddle cerebral artery in cerebrum, needling was given on Renzhong (GV 26) and Neiguan (PC 6) with electric stimulation of disperse-dense wave, 4-10 Hz frequency and 0.4 V of intensity, lasting for 5 minutes. Afterwards, pecking-acupuncture technique was done on Renzhong (GV 26) to achieve strong stimulation for 10 seconds. In shamoperation and model groups, the rats were in process of grasping and fixation, but acupuncture was not applied. Vascular endotheliocyte fluorescence staining and leukocyte fluorescence tracing method were applied, in combination with microscopic video system and computer imaging analyzing system, to observe in dynamic and quantitatively the influences of acupuncture on pia matral encephali microvascular morphology, density and blood flowing velocity at 3 hours, 6 hours and 24 hours after cerebral occlusion of meddle cerebral artery.morphology, density and blood flowing state at various time divisions observed directly microscopically and vascular endotheliocyte morphology and the state of brain tissue which near the microvascular after infusion with velocity in infarcted area at various time divisions measured in quantity in each group.microvascular endotheliocytes were colored worse and more fluorescent exudation appeared. Those were better remarkably in acupuncture group ty: That in model group was lower than normal group. That at 3 hours, 6hours and 24 hours in acupuncture group was higher remarkably than model group [(6.92±0.42), (3.25±0.52) pces/visual field; (7.61 ±0.51),(3.68±0.32) pces/visual field; (8.24±0.72), (5.49±1.2) pces/visual leukocyte velocity: That in model group was lower remarkably in normal group. That at 3 hours, 6 hours and 24 hours in acupuncture group was higher remarkably than model group [(1 193±358), (600±261) μm/s;(1 112±267), (517±115) μm/s, (1 766±293), (611±291) μm/s, q=4.608.28, P ＜ 0.01].CONCLUSION: At early stage of cerebral infarction, broken blood flow is severe in artery, vein and capillary in ischemic area; the numbers of opened vessels are few, flow rate is slow down and cerebral perfusion is unsatisfactory. Acupuncture amaliorate the microcirculation perfusion state in ischemic area, increases the numbers of opened vessels and improves flowing morphology of blood cell.

OBJECTIVETo explore protective effects of acupuncture at "Zusanli" (ST 36) on cerebral tissue in rats with sepsis.

METHODSCecal ligation and puncture (CLP) was applied to duplicate the rat model of sepsis. According to random number table, thirty SD rats were divided into a sepsis model group (group A), a sepsis model plus electroacupuncture (EA) group (group B), and a sepsis model plus non-acupoint EA group (group C), ten rats in each one. EA with the same frequency and intensity at "Zusanli" (ST 36) and non-acupoint (0.5 cm laterally to "Zusanli") for 30 min was applied in the group B and group C, respectively. No treatment was given in the goup A. 6 hours after CLP, blood was acquired from abdominal aorta to measure the levels of neuron specific enolase (NSE). Then the rats were sacrificed by abdominal aorta exsanguination to take their cerebral tissue for measuring the levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6).

RESULTSSix hours after CLP, the level of NSE was (3.51 +/- 0.39) ng/mL in group B, which was significantly lower than (7.72 +/- 0.64) ng/mL in group A (P<0.05). The level of NSE was (8.02 +/- 0.72) ng/mL in the group C, which had no statistical significance with group A (P>0.05). The levels of TNF-alpha, IL-6 in cerebral tissue in group B were significantly lower than that of group A and C (all P>0.05).

CONCLUSIONEA at "Zusanli" (ST 36) has certain protective effect on septic rat's brain, which has some relationship with decreasing levels of cerebral tissue proinflammatory cytokine and plasma NSE. EA at non-acupoint has no the same action.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Humans ; Interleukin-6 ; immunology ; Male ; Phosphopyruvate Hydratase ; immunology ; Rats ; Rats, Sprague-Dawley ; Sepsis ; enzymology ; immunology ; therapy ; Tumor Necrosis Factor-alpha ; immunology

China ; Industry ; standards ; Occupational Health Services ; standards ; Reference Standards

Aim To further investigate the protective effect and mechanisms of total paeony glycoside (TPG) on apoptosis in experimental myocardial ischemic rats.Methods The myocardial ischemic model of rats was established by subcutaneous injection of isoprenaline (ISO), and early apoptosis rate of myocardial cells and the expression of apoptosis related genes Bax, Bcl-2 and proteins in myocardial tissues were analyzed and contrasted among five groups, including normal control, ISO injury, TPG prevention and treatment, TPG treatment and positive control.Results Compared with ISO injury group, all of indexes had improved in various degrees in TPG prevention and treatment group and TPG treatment group, such as the decrease of early apoptosis rate,expression of Bax gene and protein, the increase of Bcl-2 gene and protein and the ratio of Bcl-2 protein/Bax protein.Conclusion TPG has protective effect on apoptosis in ischemic myocardium induced by ISO.The pharmacological mechanism may relate to the activation of the Bcl-2 gene and protein expression, the inhibition of Bax gene and protein expression, as well as the increase of Bcl-2/Bax ratio.

Humans ; Hyperplasia ; etiology ; Male ; Middle Aged ; Prosthesis Failure ; Stents ; Tomography, Optical Coherence ; adverse effects ; Tunica Intima ; pathology

Objective To discuss the DSA characteristics of primary hepatocellular carcinoma (HCC) and to investigate the relationship between the DSA findings and the expression of tumor’s vascular endothelial growth factor (VEGF). Methods Fifty-four patients with pathologically-proved primary HCC were enrolled in this study. The DSA findings, including tumor’s size, lesion’s number, lesion’s margin, the degree of blood supply, the presence or absence of artreiovnous shunting (AVS), etc. were observed and analyzed. The qualitative test of VEGF of HCC tissue was made by using Elivision TMplus method, and based on the stain degree and the percentage of the positive cells a comprehensive analyse was carried out. With HE stain the pathologic classification was conducted by using Edmmond method. In this study, grade I and II were regarded as well-differentiated group and grade III and IV were regarded as poorly-differentiated group. The data were processed by software SPSSS13.0 and the results were statistically analyzed. Results The expression degree of VEGF in HCC tissue was related to the blood supply pattern, the lesion’s number, the integrity of the tumor’s border and the degree of differentiation (P 0.05). Conclusion Some DSA manifestations of primary HCC bear a certain relationship to the VEGF content in the tumor tissue. Through observing the DSA characteristics, the assessment of the invasion and metastasis of HCC can be well made, which will afford the clinical physicians a useful reference in making individualized therapeutic schedule and in estimating the prognosis.

In order to protect the trial subjects' rights and benefits in drug clinical trials,we must pay special attention to the risk and benefit of the trial subjects(striving to minimize the risk and maximize the benefit),ensure the trial subjects' complete realization of the clinical trials and the successive informed consent,pay special attention to the protection of the specific trial subjects,and avoid the conflict of interests.

AlM: To investigate the relationship among carotid atherosclerosis, homocysteine ( Hcys ) and anterior ischmic optic neuropathy( AlON) .METHODS: Sixty-five AlON patients enrolled in AlON group and 70 non - AlON controls enrolled in control group. All the participants in 2 groups were examined by carotid artery color doppler flow imaging to evaluate the incidence of carotid atherosclerosis. Plasma levels of Hcys, vitamin B12 and folate were measured by automatic biochemical analyzer. The incidence of carotid atherosclerosis, plasma levels of Hcys, vitamin B12 and folate were compared statistically between AlON and control group.RESULTS: Fifty - six AlON patients ( 86%) exhibited carotid atherosclerosis, which was found in 43 control subjects ( 61%) . Fifty - nine AlON patients ( 91%) exhibited increased plasma levels of Hcys compared with that in 38 control subjects ( 54%) . Lowered vitamin B12 was found in 37 patients with AlON ( 57%) and in 43 control subjects ( 61%) . Lowered folate was found in 54 AlON patients ( 83%) and in 32 control subjects ( 46%) . The incidence of atherosclerosis in carotid artery ( P =0. 001), mean Hcys level (P=0. 0005) and lowered folate ( P = 0. 0006 ) were different statistically between AlON group and control group. Mean vitamin B12 level was indifferent between AlON group and control group ( P=0. 0722). Logistic regression analysis showed that carotid atherosclerosis was an a risk factor for AlON (OR=1. 312, 95%CI:0. 927~1. 772), whereas elevated Hcys level (OR=2. 378, 95% CI: 1. 479 ~ 3. 821 ) was also shown to be significant risk factor for AlON. CONCLUSlON: Carotid atherosclerosis and elevated Hcys were related with AlON. Reducing above-mentioned risk factors related to thrombus may be crucial to the prevention and cure of AlON.