Objective:To evaluate the antimicrobial effect of ozonated water on the putative periodontopathic bacteria.Methods:Pophyromonas gingivalis (P.g)ATCC33277,Haemophilus actinomycetemcomitans (H.a)ATCC29522,Fusobacterium nucleatum (F.n)ATCC1 0957 and clinically seperated strain of P.g(C -P.g)were treated by ozonated water with ozone concentration(mg/L) of 0.03,0.06 and 0.1 2 for 30,60,90 and 1 20 s respectively.The bactericidal effect was tested by bactericidal assay.H2 O2 was used as the positive control and distilled water as the negative control.Results:The antimicrobial rate of ozonated water agaist the bacteria increased with the ozone concentration increase.There was no statistic diffrence of the effect on P.g and C -P.g(P >0.05).Linear regression analysis showed that the βvalues of the concentration factor were over 0.95,that of the time factor under 0.1 1 .Conclu-sion:The ozonated water has dose-dependent bactericidal effect on P.g,H.a and F.n.

ObjectiveTo investigate the clinical characteristics,epidemiology of patients with severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) infection and genetic sequences of SFTSV.MethodsClinical data of five cases of severe fever with thrombocytopenia syndrome (SFTS)from Zhoushan Hospital during May 2011 to July 2011 were retrospectively analyzed.SFTSV gene was amplified by polymerase chain reaction (PCR).CD3+ CD4+ and CD3+ CD8+T lymphocytes were detected by flow cytometry (FCM).The sequences of isolated SFTSV strains were compared with those in GenBank. ResultsThe symptoms of continuous high fever,sore muscles,enlarged superficial lymph nodes,abdominal pain,diarrhea with gastrointestinal hemorrhage were observed.The white blood cells,platelets and CD3+ CD4+ T lymphocytes were progressive decreased in acute phase with the minimum of (0.97-2.00) × 109/L,(12-42) × 109/L and 7.52％-20.39％,respectively.The SFTSV was isolated from the sera of two patients.The sequences were compared with SFTSV sequences in GenBank.The homology of RNA-dependent RNA polymerase gene was 96％ compared with BX-2010,L-WWG,LN3,JS4,SD4,HN6 and AH12; the glycoprotein gene was 94％ ; N protein gene was 95％ compared with JS4,SD4 and LN4.The homology of the above three genes between two isolates was 99％.ConclusionsOur results suggest that SFTSV is sporadic in Zhejiang Province which is probably from native epidemic focus.SFTS is progressive and severe with acute onset.Multiple organ dysfunction is common in severe eases.

OBJECTIVE: To investigate the numbers, sizes and types distribution of diatoms in drowned and postmortem immersed rabbits' lungs. METHODS: Sixty-two rabbits were randomly divided into drowning group (n = 30), postmortem immersion group (n = 30) and land death group (n=2), and the diatoms in each lung lobe were analyzed quantitatively and qualitatively by microwave digestion and scanning electron microscopy. RESULTS: In the drowning group, the diatoms were detected in each lung lobe with Cyclotella and Melosira in the majority. In the postmortem immersion group, Cyclotella was in the majority. And the diatoms weren't detected in some lung lobes in postmortem immersion. There were significant differences in the detection rates of upper lobe of left lung, middle lobe and cardiac lobe of right lung in two groups (P < 0.05). CONCLUSION Based on the microwave digestion and scanning electron microscopy, the numbers, sizes and types distribution of diatoms in drowned and postmortem immersed rabbits' lungs can be analyzed and used as references for testing theory.
Animals ; Autopsy ; Diatoms/isolation & purification* ; Drowning ; Lung/microbiology* ; Microscopy, Electron, Scanning ; Microwaves ; Rabbits

BACKGROUNDTo construct recombinant retroviral vector expressing HBcAg in eukaryotic cells.

METHODSThe HBV core gene fragment was amplified by using PCR from pADR which contains complement nucleotide sequence of HBV subtype adr and cloned into retroviral expression plasmid pLXSN, then transfected into packing cell (PT67) with lipofec AMINE. After 2-3 weeks selection with G418, large colonies were isolated and transferred to individual plates. Virus-containing medium was collected and used to infect NIH3T3, EL4 and mouse bone marrow derived dendritic cells(DC). DNA was extracted from infected cells and tested by PCR. Indirect immunofluorescence and FACS were used to detect the expression of HBcAg. Cell mediated immunity of immunized C57BL/6 mice with transduced DC was analyzed.

RESULTSThe insertion of HBV core gene fragment in the recombinant retroviral plasmid was confirmed by PCR as well as enzyme digestion with EcoR1 and BamH2. The viral titer reached 3 x 10(5) CFU/ml. The result of PCR showed that the HBV core gene had been integrated into the genome of infected NIH3T3 cells. Indirect immunofluorescence and FACS analysis showed that HBcAg had been expressed in these cells. HBcAg specific CTL responses could be generated in mice immunized with retrovirus transduced DC.

CONCLUSIONSHBV core gene had been integrated into eukaryotic cells with retroviral vector and target gene had been expressed efficiently. These results may have some impact on gene therapy of chronic hepatitis B.

3T3 Cells ; Animals ; Cloning, Molecular ; Dendritic Cells ; metabolism ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; Humans ; Mice ; Recombination, Genetic ; genetics ; Retroviridae ; genetics ; Transfection

Scm in volume(42/60),multiple site involvement(44/60),blood type"O"(31/41),in comparison with those of survival group,and the difference was statistically significant.C-erbB-2,p16,p53,P-gp,CD_(44) and CD_(25)expression were not significantly different in these two groups. Conclusion The clinical stage, lymph node metastasis,lymphatic tumor emboli and/or neural involvement,infiltration depth,histological dif- ferentiation,tumor volume,involvement extension are important prognostic factors in patients with gastric can- cer,while the significance of cancer-related gene expression in gastric carcinomas needs to be studied further.

OBJECTIVETo study the difference of gene expression profiles in gastric cancer (T), pericancerous mucosa (P) and the gastric mucosa from distant cutting margin (C), and to screen an associated novel gene in early gastric carcinogenesis by oligonucleotide microarray.

METHODSU133A (Affymetrix, Santa Clara, CA) gene chip was used to detect the gene expression profile difference in T, P and C, respectively. Bioinformatics was used to analyze the detected results.

RESULTSWhen gastric cancer was compared with normal gastric mucosa, 766 genes were found,with a difference of more than four times in expression levels, including 530 up-regulated [Signal Log Ratio (SLR) > 2], and 236 down-regulated (SLR< -2). When P was compared with C, 64 genes were found, with a difference of more than four times in expression levels, including 50 up-regulated (SLR > 2), and 14 down-regulated (SLR< -2). Compared with C, a total of 143 genes with a difference of more than four times in expression levels both in T and P tissues. Of the 143 genes, 108 were up-regulated (SLR > 2), and 35 were down-regulated (SLR< -2).

CONCLUSIONSGene chip can reveal 143 same genes both in pericancerous mucosa and gastric mucosa. These genes may be related to the carcinogenesis and development of early gastric cancer.

Female ; Gastric Mucosa ; pathology ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; methods ; Precancerous Conditions ; Stomach Neoplasms ; genetics ; pathology

BACKGROUNDNontuberculous mycobacterium (NTM) had been reported to cause cutaneous infections which are difficult to interpret due to the variability of the clinical manifestations. Among NTM infections, Mycobacterium marinum (M. marinum) are mostly seen to cause skin infection. It is therefore important to establish a rapid approach for detection and identification of M. marinum from lesions of patients with suspected M. marinum infections.

METHODSSpecimens were obtained from 5 patients with swimming pool granuloma. DNA was extracted and polymerase chain reaction (PCR) was performed. PCR products were digested with Hae III and BstE II, then analysed by pattern restriction analysis to detect heat shock protein (hsp) 65 kD gene.

RESULTSThe 65 kD hsp gene was found in all specimens from patients with swimming pool granuloma. PCR restriction analysis (PRA) identified all 5 samples to be M. marinum infections, and the result was consistent with that of routine bacteriological identification. The lesions subsided or markedly improved upon treatment.

CONCLUSIONSPRA is a sensitive, specific and rapid method in identification of mycobacteria. Application of this method will be helpful for early diagnosis of mycobacterial skin infections.

Adolescent ; Adult ; Bacterial Proteins ; genetics ; Chaperonin 60 ; Chaperonins ; genetics ; Female ; Granuloma ; microbiology ; Humans ; Male ; Mycobacterium marinum ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods ; Skin Diseases, Bacterial ; diagnosis ; Staining and Labeling ; Swimming Pools

OBJECTIVETo screen the carcinogenesis associated genes in gastric carcinoma by gene chip.

METHODSU133A (Affymetrix Santa Clara, CA) gene chip was used to detect differentially expressed genes in tumor tissues, paratumor mucosa and normal mucosa. Bioinformatics was used to analyze the screened results.

RESULTSA total of 150 genes were detected with a difference of expression levels more than 3 times in paratumor mucosa compared with normal gastric mucosa, 130 of which were up-regulated and 20 down-regulated. According to the function classifications of the differentially expressed genes, the most common ones were enzyme and enzyme regulon activity associated genes(28, 18.7% ). The frequencies of nuclei acid binding activity associated genes,signal transduction associated genes and protein binding associated genes were 11.3%, 10%, and 8.7% respectively. Seventy-one differentially expressed genes were detected both in tumor tissues and paratumor mucosa compared with normal mucosa, 61 of which were up-regulated and 10 down-regulated. Among these 71 genes,e leven genes were localized on chromosome 19, 6 on chromosome 1, 2, 16, 17 respectively. No abnormal differentially expressed gene were detected on chromosome 5, 14, 22 and Y.

CONCLUSIONSThese 71 genes differentially expressed both in tumor tissues and paratumor mucosa may be associated with carcinogenesis of gastric carcinoma. The four kinds of genes associated with enzyme and enzyme regulon activity, nuclei acid binding activity, signal transduction, and protein binding should be the main genes for the study of carcinogenesis in gastric carcinoma.

Gastric Mucosa ; pathology ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Oligonucleotide Array Sequence Analysis ; Stomach Neoplasms ; genetics ; pathology

OBJECTIVETo study the biological characteristics of cytotoxicity T lymphocyte (CTL) induced by dendritic cell (DC) transfected with the Epstein-Barr virus latent membrane protein 2A (EBV-LMP2A) recombinant adenovirus. To establish nasopharyngeal carcinoma (NPC) animal models expressing LMP2A and investigate the anti-tumor effect of LMP2A specific CTL in vivo.

METHODSThe mononuclear cells were isolated from human peripheral blood mononuclear cells (PBMC) and cultured with the cytokines [granulocyto-monocyte colony stimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor-alpha TNF-alpha]. The expression of surface markers on mature DC was detected by fluorescence activated cell sorter FACS. Mature DC were transfected with LMP2A recombinant adenovirus. Under the help of interleukin-2 (IL-2), LMP2A specific CTL were induced by coculturing LMP2A-transfected DC with autologous PBMC. The population of CTL was detected by FACS. NPC animal models were constructed by implanting CNE cells expressing LMP2A subcutaneously into BALB/c nude mice. After intra-tumoral injection of LMP2A specific CTL, the size of tumor was measured. The tumors were removed after 30 d and subjected to histological examination.

RESULTSMature DC displaying typical characteristics of morphology and phenotype were obtained from monocytes cultured in the medium containing GM-CSF, IL-4 and TNF-alpha. The LMP2A specific CTL induced by transfected DC were composed of mainly CD4+ and CD8+ T cells. The NPC animal models were constructed three weeks after implanting CNE cells. The study in vivo indicated that the tumors treated with LMP2A specific CTL grew slowly compared with control. Tumor volume of treated groups was significantly smaller than that of controls. The histological sections showed local necrosis and infiltration of lymphocyte in tumor tissue.

CONCLUSIONSTypically mature DC could be generated in vitro by culturing monocytes with the cytokines. LMP2A-specific CTL could be induced by LMP2A transfected DC in vitro. NPC mice models could be constructed by implanting CNE cells. LMP2A specific CTL could inhibit the growth of implanted tumor and produce an anti-tumor effect in vivo.

Adenoviridae ; genetics ; Animals ; Cell Line, Tumor ; Dendritic Cells ; immunology ; Female ; Herpesvirus 4, Human ; Humans ; In Vitro Techniques ; Mice ; Mice, Inbred BALB C ; Nasopharyngeal Neoplasms ; pathology ; therapy ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Viral Matrix Proteins ; genetics

OBJECTIVETo assess the impact of the heat wave in 2005 on mortality among the residents in Guangzhou and to identify susceptible subpopulations in Guangzhou, China.

METHODSThe data of daily number of deaths and meteorological measures from 2003 to 2006 in Guangzhou were used in this study. Heat wave was defined as ⋝7 consecutive days with daily maximum temperature above 35.0 °C and daily mean temperature above the 97th percentile during the study period. The excess deaths and rate ratio (RR) of mortality in the case period compared with the reference period in the same summer were calculated.

RESULTSDuring the study period, only one heat wave in 2005 was identified and the total number of excess deaths was 145 with an average of 12 deaths per day. The effect of the heat wave on non-accidental mortality (RR=1.23, 95% CI: 1.11-1.37) was found with statistically significant difference. Also, greater effects were observed for cardiovascular mortality (RR=1.34, 95% CI: 1.13-1.59) and respiratory mortality (RR=1.31, 95% CI: 1.02-1.69). Females, the elderly and people with lower socioeconomic status were at significantly higher risk of heat wave-associated mortality.

CONCLUSIONThe 2005 heat wave had a substantial impact on mortality among the residents in Guangzhou, particularly among some susceptible subpopulations. The findings from the present study may provide scientific evidences to develop relevant public health policies and prevention measures aimed at reduction of preventable mortality from heat waves.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Female ; History, 21st Century ; Hot Temperature ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Mortality ; Weather ; Young Adult