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

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

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 study the mechanism underlying the effect of combined use of cyclonpamine and hydroxycamptothecin in inducing the apoptosis of human oral squamous cell carcinoma cell line (OSCC) HSQ-89.

METHODSCCK8 assay was used to investigate the inhibitory effect of cyclopamine on HSQ-89 cells. Flow cytometry (FCM) was employed to examine the cell apoptosis following combined treatment with cyclonpamine and hydroxycamptothecin. Reverse transcription polymerase chain reaction (RT-PCR) was applied to detect the mRNA expressions of Bcl-2, Bcl-xl, and Bid in HSQ-89 cells after the treatments.

RESULTSCombined treatment with cyclonpamine and hydroxycamptothecin significantly inhibited the cell proliferation compared with hydroxycamptothecin treatment alone, also resulting in a significantly higher apoptosis rate of the cells (P<0.05). The mRNA level of Bcl-2 was significantly decreased after the treatments, especially after the combined treatment. Cyclopamine produced no significant effect on the mRNA levels of Bcl-xl and Bid in the cells.

CONCLUSIONThe combined use of cyclopamine and hydroxycamptothecin significantly down-regulates the expression on of bcl-2 to induce the apoptosis of human OSCC cell line HSQ-89.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Camptothecin ; analogs & derivatives ; pharmacology ; Carcinoma, Squamous Cell ; pathology ; Cell Line, Tumor ; Drug Synergism ; Humans ; Mouth Neoplasms ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Veratrum Alkaloids ; pharmacology

BACKGROUNDSchwannomatosis is a recently recognized peripheral nerve polyneoplasm with clinical characteristics and a genetic background that differ from those of neurofibromatosis 2 (NF2). The diagnostic and treatment criteria of this rare disorder are herein discussed.

METHODSThe data of 180 patients who underwent operations for benign schwannomas from 2003 to 2012 in our center were reviewed. Eight of them were classified as schwannomatosis according to the diagnostic criteria suggested by MacCollin. The demographic characteristics were documented and compared between the two groups of patients. The patients' clinical presentations, imaging characteristics, histological features, and treatment results were retrospectively investigated and summarized.

RESULTSOf the 180 cases of benign schwannomas we reviewed this time, eight patients presented with schwannomatosis (4.44%). The mean age of the two groups was not significantly different (40.0 vs. 44.7 years, t = 0.88, P = 0.378). However, schwannnomatosis seems to more generally occur in females (75% vs. 48% were females, P = 0.162), although the difference was not statistically significant. The initial main symptom was pain. The neurological examination was otherwise normal. Magnetic resonance imaging (MRI) revealed multiple discrete, well-defined round, or oval lesions distributed along the course of the peripheral nerves in the extremities with low-to-intermediate signal intensity on T1-weighted images and high-signal intensity on T2-weighted images. Vestibular schwannomas were excluded in four patients by cranial MRI. The lesions in all patients were resected and were pathologically proven to be schwannomas. The average follow-up period was 26 months. Six individuals obtained a good result without symptoms or function loss.

CONCLUSIONSSchwannomatosis is characterized by the development of multiple schwannomas without evidence of the vestibular tumors that are diagnostic for NF2. It commonly occurs in middle-aged females. It has similar demographic features to solitary benign schwannoma. Surgical resection always results in a good outcome.

Adult ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neurilemmoma ; genetics ; pathology ; surgery ; Neurofibromatoses ; genetics ; pathology ; surgery ; Skin Neoplasms ; genetics ; pathology ; surgery