Adolescent ; Child ; Child, Preschool ; Female ; Gastric Mucosa ; pathology ; Gastritis ; pathology ; Humans ; Male ; Metaplasia ; Retrospective Studies

OBJECTIVETo establish and optimize the two dimensional gel electrophoresis (2-DE) map of epidermal cells separated from the hypertrophic scar tissues so as to explore the function of the non cultured epidermal cells during the course of the formation of hypertrophic scar.

METHODSTo separate the epidermal cells from the scar tissues, the scar epidermis was digested with Dispase II and trypsin. The total protein of the cells was then extracted and separated with 2-DE and visualized with silver stain. Spots detection and matching were performed with Melaine 3.0 gels analyzing software. The results were then compared with the normal epidermal cells' 2-DE map coming from the Danish Center for Human Genome Research' s 2-DE PAGE Databases.

RESULTSNearly more than 600 protein spots were identified in the final optimized 2-DE map. We found out 24 differentially expressed proteins by comparing the difference in composition, shape or density of all the spots. In the 24 proteins, there are 8 up-regulated ones, 9 down-regulated ones, 4 disappeared ones and 3 newly founded ones.

CONCLUSIONSThe method of digesting the epidermis with Dispase II and trypsin to separate the epidermal cells to establish the 2-DE map is feasible and it made the further study on hypertrophic scar proteomics possible. The 24 differentially expressed proteins revealed that epidermal cells might play a role in the formation of hypertrophic scar.

Cell Line ; Child ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; methods ; Epidermis ; metabolism ; Humans ; Proteome ; metabolism

OBJECTIVETo investigate the relationship between the resuscitation promoting role of resuscitation promoting factor and the initial bacteria amount of dormant Mycobacterium tuberculosis.

METHODSMycobacterium tuberculosis (dormant bacteria) was cultured for 100 days, then diluted into 1 mg/ml concentration with 7H9, and further diluted into 0.5, 0.25, 0.125, 0.0625, and 0.03125 mg/ml. Twelve new tubes added with 5 ml 7H9 and divided into two groups: the first group was added with the resuscitation-promoting factor protein, and the second group as control was added with 7H9. In each group the above diluted solutions were added. The tubes were located at 37 degrees C for culture. Optical density (OD) was detected on day 15, 25, 30, and 35. From each tube 1 microl culture solution was plated on 7H11 medium for colony counting.

RESULTSOD detection showed that bacteria proliferation in each group had positive linear correlation (P < 0.05, P < 0.01), indicating that the resuscitation-promoting factor played a similiar role in solutions with different dilution concentrations. 7H11 results and the OD results show that these two detection methods in each group had linear correlation (P < 0.05, P < 0.01), indicating that these two methods showed consistent test results.

CONCLUSIONThe resuscitation-promoting factor has no effect on the resuscitation of dormant Mycobacterium tuberculosis and its initial bacteria amount.

Bacterial Proteins ; metabolism ; Cytokines ; metabolism ; Mycobacterium tuberculosis ; physiology ; Resuscitation

OBJECTIVETo establish a rapid, inexpensive, and simple drug susceptibility test (DST) for Mycobacterium tuberculosis (M. tb) and evaluate its feasibility.

METHODWe used nitrate reductase combined with mycobacteriophage assay (PhaB-NRA) to test 49 clinical M. tb isolates of, and the results were compared with those of PhaB-NRA and traditional absolute concentration method.

RESULTSThe sensitivity, specificity, and accuracy of PhaB-NRA for rifampicin were 89.1%, 91.67%, and 89.8%; on the contrary, those of isonicotinyl hydrazide were 86.21%, 90.0%, and 87.8%, respectively. The coincidence between PhaB-NRA and traditional assay were 0.746 for rifampicin and 0.750 for isonicotinyl hydrazide.

CONCLUSIONSPhaB-NRA is an inexpensive, rapid, and simple DST method. It is a promising rapid screening technique for DST of M. tb.

Anti-Bacterial Agents ; pharmacology ; Biological Assay ; methods ; Humans ; Microbial Sensitivity Tests ; methods ; Mycobacteriophages ; physiology ; Mycobacterium tuberculosis ; drug effects ; Nitrate Reductase ; metabolism ; Rifampin ; pharmacology ; Sensitivity and Specificity

OBJECTIVETo develop an experimental rat model of inflammatory bowel disease (IBD) by administration of dextran sulfate sodium (DSS), and to observe changes in the tight junction protein expression and permeability of colon mucosa.

METHODSMale Sprague-Dawley (SD) rats were randomly divided into control (n=27) and IBD model groups (n=27). In the IBD model group, IBD was induced by 6-day administration of 3% DSS in water followed by 14-day administration of water only. The control group was fed with water only. Pathological changes in colon mucosae were observed on days 7, 14 and 21 after DSS administration. Colon tissue specimens were collected on day 21 for measuring myeloperoxidase (MPO) activity. The transepithelial electric resistance (TEER), transepithelial potential difference (TEPD) and short circuit current (Isc) of the specimens were measured by Ussing chamber. Real-time PCR and Western blot were used to measure the mRNA and protein expression of tight junction proteins in colon epithelia.

RESULTSIn the IBD model group, diarrhea, hemafecia and weight loss were seen. Inflammation occurred mainly in the distal colon and was characterized by crypt abscess and inflammatory cell infiltration. The IBD model group showed significantly increased MPO activity (P<0.01), significantly decreased TEER (P<0.01) and TEPD (P<0.01), and significantly increased Isc (P<0.01) compared with the control group. No claudin 2 expression of mRNA and protein was detected in the control group, and they were expressed in the IBD model group. The expression levels of claudin 3, occludin and ZO-1 in the IBD model group were significantly decreased compared with in the control group (P<0.01).

CONCLUSIONSIBD rats show colonic barrier dysfunction and changes in the expression of tight junction proteins. The changes in the expression of tight junction proteins may contribute to colonic barrier dysfunction in cases of IBD in the chronic recovery stage.

Animals ; Claudin-3 ; analysis ; Colon ; metabolism ; pathology ; Dextran Sulfate ; Disease Models, Animal ; Inflammatory Bowel Diseases ; chemically induced ; metabolism ; Intestinal Mucosa ; metabolism ; Male ; Occludin ; analysis ; Permeability ; Rats ; Rats, Sprague-Dawley ; Tight Junction Proteins ; analysis ; Zonula Occludens-1 Protein ; analysis

The standardized training is an indispensible stage for the improvement of residents' comprehensive quality and for the training of high-qualified talents.The article preliminarily explored the standardized training model for residents,which was in accordance with the characteristics of the department of cardiology mainly from four aspects:the set-up of reasonable training program,the training of practical skills,the training of humanistic quality and the training of life-long learning ability.

OBJECTIVETo observe the correlation between brain-derived neurotrophic factor (BDNF) expression in the nerve system of diabetes mellitus (DM) rats and diabetic erectile dysfunction (ED).

METHODSDM rats were induced by injecting streptozotocin and erectile function test was done by injecting apomorphine (APO) at 1 month, 2 months, 3 months and 4 months. Then the brain, lumbosacral spinal cord, thoracic and lumbar sympathetic trunks, penis and prostate were taken from the diabetic and normal rats of the same age. The BDNF positive neurons and nerve fibers were shown by immunohistochemistry or fluorescence immunohistochemistry. The number and the grey density of BDNF positive cells and fibers were detected by image analysis.

RESULTSCompared with the control group, the erection frequency of the DM rats decreased at 2 months (P <0. 05) , and significantly at 3 and 4 months (P > 0.01) , and the BDNF positive neurons and nerve fibers in the cerebral cortex, lumbosacral spinal cord, thoracic and lumbar sympathetic trunks, penis and prostate of 1-month DM rats were reduced (P <0. 05). As time went on, BDNF declined progressively.

CONCLUSIONBDNF decreases in the central and peripheral nerve system in the early stage of diabetes mellitus, wich is closely correlated with diabetic ED.

Animals ; Brain-Derived Neurotrophic Factor ; biosynthesis ; Diabetes Mellitus, Experimental ; physiopathology ; Erectile Dysfunction ; physiopathology ; Male ; Nerve Tissue ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo measure the vertical height of mesio-distal marginal ridge to cusp in posterior teeth, which may be helpful to brackets positioning.

METHODSThe study groups comprised of 60 patients (30 men, 30 women, mostly aged 12-14 years) who underwent orthodontic treatment without tooth extraction and matched the Andrews normal occlusion standard after treatment. Study model of each patient was made. Three-dimensional laser measurer was used to evaluate the vertical height of mesio-distal marginal ridge to mesial cusp in posterior teeth. The data were stored in a personal computer and submitted to statistical analysis of paired t test.

RESULTSNo statistical significant difference was found in the same teeth between men and women. Not only in maxilla but also in mandible, there was no significant difference between the left and the right (P>0.05). The average vertical height of maxillary first premolars was (1.70+/-0.50) mm, the maxillary second premolars was (1.24+/-0.45) mm, and for maxillary first molars, the result was (0.83+/-0.40) mm. The difference between each result was statistically significant (P9< 0.01). The average vertical height of mandibular first premolars was (2.25+/-0.45) mm, the mandibular second premolars was (1.55+/-0.45) mm, and for mandibular first molars, the result was (1.18+/-0.40) mm. The difference between each result was statistically significant (P<0.0 1).

CONCLUSIONThe vertical height of brackets position in posterior teeth should be considered to guarantee that mesio-distal marginal ridges of deferent posterior teeth located in the same plane, so that satisfying goal could be achieved, If the vertical height in the first molar was X mm, the vertical height in the second premolar should be (X+0.5) mm, and (X+1.0) mm might be suit for the first premolar.

Bicuspid ; Dental Occlusion ; Female ; Humans ; Male ; Mandible ; Maxilla ; Molar ; Tooth ; Tooth Extraction ; Tooth Movement Techniques

OBJECTIVETo analyze characteristics of different hyperphenylalaninemia (HPA) and to discuss the clinical difference between southern and northern Chinese patients with tetrahydrobiopterin (BH4) responsive phenylalanine hydroxylase (PAH) deficiency.

METHODS(1)BH4 (20 mg/kg) loading test was performed in all 108 HPA patients. These patients, 63 males and 45 females, were at a mean age of 7.05 months. A combined phenylalanine (Phe) and BH4 loading test was carried out in the patients who had a basic blood Phe concentration less than 600 micromol/L. The urine pterine profile analysis and the dihydropteridine reductase (DHPR) activity in dry blood filter spot were analyzed simultaneously. (2)BH4 responsive patients were divided to southern and northern groups by their parent's native place and geographic boundary determined by Changjiang River. The change of Phe concentration after BH4 loading test was compared between the two groups.

RESULTS(1)Among the 108 HPA cases, 36 patients (33.3%) were BH4 responsive PAH deficiency, 49 (45.4%) were non-BH4 no responsive phenylketonuria (PKU)and 23(21.3%)were BH4 deficiency (BH4D). The Phe concentration of patients with BH4 responsive PAH deficiency decreased by 49.24% and 65.35% at 8 h and 24 h after oral BH4, 23 in southern group and 13 in northern group among 36 patients. (2)The mean Phe concentration at 24 h after loading test in southern and northern groups were (217.02+/-189.03) micromol/L and 458.75+/-342.54 micromol/L respectively (P<0.05), although the decrease percent of plasma Phe concentration at 2 h, 4 h, 8 h, 24 h was no distinct difference between southern and northern groups (P>0.05).

CONCLUSIONMost of mild and moderate HPA patients affected by PAH deficiency show plasma Phe concentration decrease >30% in 24 h after oral BH4 20 mg/kg, few are classic PKU. The responsiveness to BH4 is no difference between southern and northern Chinese patients with BH4 responsive PAH deficiency according to the decrease percent of plasma Phe concentration, although the Phe concentration is lower in southern patients than that in northern patients.

Biopterin ; analogs & derivatives ; pharmacology ; therapeutic use ; Child, Preschool ; China ; Dihydropteridine Reductase ; blood ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Phenylalanine ; blood ; Phenylketonurias ; blood ; drug therapy ; Time Factors

OBJECTIVETo develop a newly real-time RT-polymerase chain reaction assay for severe acute respiratory syndrome (SARS) related coronavirus in human whole blood.

METHODSA pair of primers and a probe (molecular beacon) had been designed that were specific for the recognition of a highly conservative region between 15 301 and 15 480 of the SARS-related coronavirus polymerase gene sequences obtained from GenBank (G130027616).

RESULTSIn the real-time RT-PCR assay, the extent of SARS related coronavirus amplification was measured in terms of the increase in fluorescence during the amplification process. The 145 bp fragment of PCR product was further confirmed by conventional PCR assay and proved by DNA sequencing to be identical to the target sequence to which the probe was hybridized.

CONCLUSIONThis assay has a broad application for clinical diagnosis and surveillance investigation.

Base Sequence ; Humans ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; Severe Acute Respiratory Syndrome ; diagnosis