Objective:To evaluate the physiological roles of bmp3,bmp4 and bmp7 during the formation of permanent tooth roots in dog. Methods:The expression of bmp3,bmp4 and bmp7 mRNA at different stages of the development of permanent tooth roots was examined by in situ hybridization in 3 dogs aged 12-18 weeks. Results:bmp3 was found in dental sac surrounding the germs at the early stage of tooth root development, and in cementoblasts and periodontal cells at the later stage. bmp4 was found in odontoblasts, dental papilla and osteoblasts. bmp7 positive signals was found only in epithelial cells of root sheath around cervical circulus at early stage, then located in cementoblasts and odontoblasts at later stage. Conclusion:The spatiotemporal expressions of bmp3,bmp4 and bmp7 are widely diverse, indicating that they participate in the regulation of tooth root development.

Objective To evaluate the clinical effects of ultrasound-guided internal jugular vein catheterization in long axis plane,short axis plane and oblique axis plane,in order to identify the opti-mal axis plane for this procedure.Methods One hundred and eighty patients (male 94 cases,female 86 cases,aged 34-82 years)requiring ultrasound-guided internal jugular vein catheterization were in-cluded in this study.They were randomly divided into three groups (n =60 each),long axis group, short axis group and oblique axis group,with 60 cases in each group.The details of catheterization in-cluding the time accessing into vein,the time finishing cannulation,needle redirecting times,number of skin points of puncture,puncture successful rate and complications in the three groups were recor-ded.Results Compared with long axis plane and short axis plane,the oblique axis plane was associat-ed with decreased time for venous access and cannulation.The oblique axis plane also needed less changes of needle direction.The complication of arterial puncture in the oblique axis plane group was significantly lower than long axis plane group and short axis plane group(P <0.05).The number of skin puncture points were similar between the three groups.Conclusion The oblique plane can provide a safe and more effective route to perform the IJV catheterization with minimal risk for carotid artery puncture,which demonstrates the practical superiority over the classic short axis plane and long axis plane for critically ill patients.

To improve the efficiency of halophilic actinobacteria screening and carry out the rapid selection of targeted strains, we tested 34 strains of Streptomonospora by PCR-single strand conformation polymorphism analysis (PCR-SSCP) based on genus-specific primers for the PCR identification. This approach employs PCR with two pairs of primers located in the 16S rRNA sequence flanking two variable region, then build clustering tree according as SSCP data. Synchronously, we sequenced all the 16S rRNA partial sequences for these strains to verify them. The results showed that the PCR-SSCP analysis was an efficient, easy-to-handle and economic method for rapid selection of halophilic actinobacteria resources.

OBJECTIVETo explore the effect of Yiqi Huoxue recipe (YHR), a Chinese herbal medicine for supplementing Qi, activating blood circulation to remove stasis, on vascular extracellular matrix (ECM) remodeling and its molecular mechanism.

METHODSVascular smooth muscle cell (VSMC) proliferation activity and collagen turnover rate were detected by 3H-TdR test and hydroxyproline amount determined by 3H-Pro incorporation. Expression activity of MMP-2 and osteopontin genes was detected by Northern blotting and MMP-2 zymography analysis.

RESULTSYHR could markedly inhibit VSMC collagen synthesis stimulated by blasic fibroblast growth factor (bFGF) and lower the collagen turnover rate induced by vascular de-endothelialization. The expression level of MMP-2 and osteopontin genes was down-regulated by YHR in cultured VSMC and vascular wall with endothelial injury, and VSMC proliferation was inhibited by the serum obtained from YHR treated rats. Removing protein from the drug serum made no change on the effect of YHR to VSMC.

CONCLUSIONYHR could inhibit and/or retard ECM remodeling through regulating the expression of MMP-2 and osteopontin genes and lowering the collagen turnover rate.

Animals ; Aorta ; cytology ; Cell Division ; drug effects ; Cells, Cultured ; Collagen ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Matrix ; genetics ; metabolism ; Fibroblast Growth Factor 2 ; metabolism ; Gene Expression ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Osteopontin ; Rats ; Rats, Sprague-Dawley ; Sialoglycoproteins ; genetics ; metabolism

AIM AND METHODSTo determine the relationship between the nuclear envelope nucleoside triphosphatase (EC 3. 6. 1. 15, NTPase) activity and the phenotypic modulation of vascular smooth muscle cell (VSMC), the NTPase activity was detected during restenosis after de-endothelialization in vascular wall. The activities of three enzymes involved in carbohydrate and nucleic acid metabolism were also investigated by spectrophotometry.

RESULTSThe activity of NTPase increased continuously and associated with the process of intimal thickening. Western blotting showed that expression of SMalpha-actin, as the marker of contractile phenotype of VSMC, decreased continuously. Osteopontin (OPN), the marker of synthetic phenotype of VSMC, was up-regulated during the process. These suggested that intimal injury induced phenotypic modulation of VSMC. The activities of 5'-nucleotidase, adenosine deaminase and succinate dehydrogenase increased and reached their peaks on 7 days after de-endothelialization. The changes of three enzymes were associated with proliferation in VSMC.

CONCLUSIONThe efflux of mRNA and the changes of enzyme activity involved in carbohydrate or nucleic acid metabolism may be the biochemical basis in the development and progression of restenosis.

Animals ; Constriction, Pathologic ; Endothelium, Vascular ; pathology ; Female ; Hyperplasia ; enzymology ; pathology ; Male ; Muscle, Smooth, Vascular ; pathology ; Nucleoside-Triphosphatase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tunica Intima ; enzymology ; pathology

Objective To investigate the effects of different extracts of pericarpium citri reticulatae (PCR) and pogostemon cablin benth (PCB) on the contraction of gastrointestinal smooth muscle and the level of gastrointestinal hormones in rat model of gastrointestinal motility disorder. Methods Seventy Wistar rats were randomly divided into groups of control, model, water extractive of PCR, hesperidin, water extractive of PCB, water extractive of PCB plus volatile oil and patchouli alcohol, ten rats in each group. Except the control group, the rest groups were established gastrointestinal motor disorder model via limb ischemia-reperfusion (LIR). After modeling rats of groups were intervened with corresponding extracts. The effects of different extracts on contraction amplitude of corpora ventriculi and small intestine smooth muscle were observed. The levels of gastrointestinal hormones including motilin (MOT), gastrin (GAS), cholecystokinin (CCK) and somatostatin (SS) were detected by radioimmunoassay. Results The contraction amplitudes of corpora ventriculi and small intestine smooth muscle were decreased (P<0.05), the serum level of GAS and plasma level of MOT were also significantly decreased, while CCK and SS levels in the gastric antrum were significantly increased in model group than those of the control group (P < 0.05). Water extractive of PCR, hesperidin, water extractive of PCB and water extractive of PCB +patchouli oil can increase the contraction amplitudes of corpora ventriculi and small intestine smooth muscle, increase the serum level of GAS and reduce levels of CCK and SS in the gastric antrum (P<0.05), whereas showed no influence in the plasma level of MOT (P>0.05)]. Compared with model group, patchouli alcohol showed no influence in the contraction of gastrointestinal smooth muscle and levels of MOT, GAS, CCK and SS (P>0.05). In the aspect of regulating the contraction of gastrointestinal smooth muscle and the level of GAS, CCK and SS, the pharmacological effect of PCR water extract was better than that of hesperidin (P<0.05), while water extractive of PCB+volatile oil was better than that of water extractive of PCB (P < 0.05). Conclusion The active ingredients of PCR and PCB have variant regulative effects on the contraction of gastrointestinal smooth muscle and the serum level of GAS, CCK and SS in the gastric antrum in rat model of gastrointestinal motility disorder.

OBJECTIVETo prepare and characterize monoclonal antibodies (mAb) and polyclonal antibodies against nucleocapsid (N) protein of severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) and to establish antibodies-based sandwich ELISA for detecting N protein of SARS-CoV, which might apply to early diagnosis of patients with SARS-CoV infection.

METHODSBALB/c mice were immunized with purified recombinant N protein of SARS-CoV for producing mAbs, and New Zealand white rabbits were immunized for producing polyclonal antibodies. The identification of antibodies was performed using indirect enzyme-linked immunosorbent assay (ELISA), indirect fluorescent-antibody assay (IFA), and Western immunoblotting. Capturing and detecting antibodies were selected by pairing the mAbs and polyclonal antibodies one by one and an antibodies-based sandwich antigen capture ELISA was used for detecting N antigen of SARS-CoV.

RESULTSNine mAbs and hyperimmune rabbit polyclonal antibodies, specifically against SARS-CoV nucleocapsid protein were obtained. Using paired ELISA assay, three mAbs N1E8, N8E1 and N10E4 were selected as capturing antibody and rabbit polyclonal antibodies as detecting antibody then triple antibodies-based sandwich ELISA was established following horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G. The recombinant N protein was used as a standard to establish a detection sensitivity of approximated 50 pg/ml with this assay. When tested with 420 serum specimens from serologically confirmed SARS patients, the positive rates of serum N protein were 90.1%, 23% and 0%, in which sera collected from 1 to 10 days, 11 to 20 days and beyond 21 days respectively after the onset of symptoms. The specificity of the assay was 99.86% in 715 control serum specimens. There was no cross-reaction with other respiratory viruses and coronaviruses.

CONCLUSIONSpecific and high affinity mAbs and rabbit polyclonal antibodies were obtained. By paired and optimized sandwich ELISA, a sensitive and specific antigen capture ELISA was established for detecting N antigen of SARS-CoV, which might apply to early diagnosis, source tracing and epidemiological studies of SARS.

Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibodies, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; Humans ; Mice ; Mice, Inbred BALB C ; Nucleocapsid ; immunology ; Rabbits ; SARS Virus ; immunology ; isolation & purification ; Sensitivity and Specificity ; Severe Acute Respiratory Syndrome ; virology

OBJECTIVETo observe the feasibility of measuring multiple circulating cytokines by protein chips in patients with acute coronary syndrome (ACS).

METHODSCirculating cytokines were determined by protein chips in 50 control cases and 104 ACS cases of unstable angina pectoris (UA, n = 50) and acute myocardial infarction (AMI, n = 54) cases.

RESULTSThe levels of serum MMP-9, sCD40L, CRP, IL-6, H-FABP, cTnI and plasma IL-8, NT-proBNP, sVCAM-1, ET-1 in ACS group were significantly higher than those in the control group (all P < 0.01). Serum H-FABP and cTnI were significantly higher in AMI patients than in UA patients (P < 0.01). Serum MMP-9, sCD40L, CRP, IL-6 and plasma sVCAM-1, ET-1 tended to be higher in AMI patients as compared to UA patients (all P > 0.05). Serum MMP-9, sCD40L, and H-FABP were positively correlated with the severity of angina(r = 0.653, r = 0.745, r = 0.933, P < 0.01, respectively). Serum MMP-9, sCD40L, and H-FABP increased in proportion to the levels of Braunwald angina classes in UA patients (angina class I < angina class II < angina class III, P < 0.01).

CONCLUSIONSCirculating cytokines were significantly increased in ACS patients and positively correlated to Braunwald angina classes in UA patients.

Acute Coronary Syndrome ; blood ; Aged ; Angina Pectoris ; blood ; Angina, Unstable ; blood ; C-Reactive Protein ; metabolism ; CD40 Ligand ; blood ; Cytokines ; blood ; Female ; Humans ; Male ; Matrix Metalloproteinase 9 ; blood ; Middle Aged ; Myocardial Infarction ; blood ; Protein Array Analysis

The objective visual acuity test is mandatory in certain cases, such as infants, nonverbal subjects and subjects who need legal judgements. To produce more reliable objective method, we made a new objective system for visual acuity test which is called Seohan visual acuity test(SVT). The SVT system has three elements such as stimuli, display and evaluation. For the visual stimuli, the computer program for optokinetic nystagmus(OKN)test was set up in personal computer to control the size, speed and direction of the stimuli easily. The visual stimuli are presented on HeadMounted-Display(HMD)to separate the stimuli from the environment to keep the uniform size and distance of screen to the eye of subjects and project the stimulus in full central field of patient. Electrooculography(EOG) was used to assess the visual acuity more objectively. To evaluate the usefulness of SVT, the smallest size of the stimulus to elicit OKN(objective visual acuity)with SVT was obtained in 10 eyes in each twelve grade of subjective visual acuity(decimal V). Objective visual acuity(VA)were distributed from 8.3+/-1.87 to 21.4+/-3.1 and showed high correlation(p<0.00 ). The relationship between the objective VA obtained from SVT and subjective VA make a regression line(y=-12.874X+21.303).Subjective VA could be obtained from conversion of objective VA with 95%confidence belt. In conclusion, the objective visual acuity with SVT is highly correlated with subjective visual acuity and SVT using the OKN response to stimuli presented on HMD by the computer program can be useful in assessing visual function objectively.
Electrooculography ; Humans ; Infant ; Microcomputers ; Nystagmus, Optokinetic* ; Visual Acuity*

Several tests have been used for assessing color vision but most of them are nonspecific, expensive, complicated or too time consuming to perform. We have developed a new computerized color vision test named Seohan computerized 85-hue test[SCHT/85]that has many advantages compared to Farnsworth Munsell[FM]100-hue test.SCHT/85 has 85 color caps which can be displayed based on a model of FM 100-hue test using Multimedia toolbook 4.0.Selected color caps in the lower random area can be moved and inserted to the upper result area by a mouse.At the end of the test, error scores and ring graph can be displayed in monitor and can be also printed out. Total 15 subjects performed with SCHT/85 and FM 100-hue test twice who have been showed 10 normal and 5 abnormal color vision reported with other color vision tests and the results of two tests were analysed to compare the total error scores[TES]and reproducibility. All ten normal subjects showed lower TES in SCHT/85 than FM 100-hue test and the mean TES were 31.5 and 43.6 with SCHT/85 and FM 100-hue test.The coefficients of variation [CV]were 4.3%and 22.7%with SCHT/85 and FM 100-hue test.The two tests were highly correlated[r=0.92].Five abnormal subjects showed higher TES in SCHT/85 than FM 100-hue test and the mean TES were 169.8 and 157.3 with SCHT/85 and FM 100-hue test.The coefficients of variation were 9.1%and 21.2%with SCHT/85 and FM 100-hue test.These two tests were also highly correlated[r=0.965]. In conclusion, SCHT/85 which was newly developed by authors is a highly useful test for color vision.
Color Vision* ; Multimedia ; Pilot Projects*