Many psychrophilic bacteria were isolated from east-pacific sediment which is 5,300m in depth. The genera of 13 isolations were identified and classified based on the comparison and phylogenetic study of 16S rRNA gene sequences. The results indicated that they belong to four genera including Parococcus, Pseudomonas, Halomonas and Pseudoalteromonas.

Objective To develop a mouse model of cutaneous protothecosis.Methods Totally,48 BALB/c mice were randomly and equally divided into four groups:high-and low-concentration immunosuppressive groups-immunosuppressed mice inoculated with P.zopfii var.portoricensis suspension of 1 × 109 and 1 × 106 colony forming units (CFU) conidia/ml respectively,high-concentration healthy group-healthy mice inoculated with P.zopfii var.portoricensis suspension of 1 × 109 CFU conidia/ml,and control group-healthy mice inoculatedwith sodium chloride physiological solution.The P.zopfii suspension or sodium chloride physiological solution was subcutaneously inoculated to the abdominal skin of mice,with the inoculation volume being 200 μl.Skin appearance at the inoculation site was observed,and four mice were sacrificed in each group on day 7,14 and 28 after the inoculation.Skin specimens were resected from the inoculation sites of mice and subjected to pathological and mycological examinations.Results Mice in the three experiment groups inoculated with the P.zopfi suspension were all infected,with the appearance of papules and abscess at the inoculation sites of all mice as well as ulcer and crusts in some mice.Meanwhile,no mice were infected in the control group.Significant differences were noted in the diameter of skin lesions between the three time points in these experiment groups (all P ＜ 0.05),and the largest diameter of lesions was observed on day 7,which was (6.75 ± 1.09) mm in the high-concentration immunosuppressive group,(5.88 ± 1.17) mm in the low-concentration immunosuppressive group,and (5.96 ± 0.99) mm in the high-concentration healthy group.Comparisons of lesion diameter between the three experiment groups revealed a statistical difference on day 28 (F =8.91,P ＜ 0.05),with the largest lesion diameter observed in the high-concentration immunosuppressive group (4.38 ± 0.86 mm),but no statistical difference was found on day 7 or 14 (both P ＞ 0.05).Pathology of skin specimens consistently revealed necrosis,abscess and granuloma formation in the three experiment groups.Spores were found in the lesions of infected mice by haematoxylin-eosin staining,periodic acid-Schiff staining,and direct microscopy.Culture of tissue samples from the experiment groups grew Prototheca.Conclusion The mouse model of protothecosis can be established by subcutaneous inoculation of Prototheca suspension in the abdominal skin of healthy or immunosuppressed mice.

Objective To investigate the apoptotic effects of hypertrophic scar fibroblast (HSF) induced by HMME-PDT.Methods Fibroblasts were cultured from nontreated hypertrophic scars,and cells at passages 4-6 were used for the experiments (photosensitizer dose 4 μg/ml,λ630 nm,pow er density 10 mw/cm2,energy fluence 2.5 J/cm2).Morphological and biochemical changes in fibroblasts were assessed by Hoechst 33258 staining and fluorescence microscopy.The rate of apoptotic or necrotic cells was detected by flow cytometry (FCM) through double staining of Annexin V -FITC and popodium iodide (PI),respectively.Results Marked morphological features of cell apoptosis were viewed under the fluorescent microscope through Hoechst 33258 staining.The analysis of FCM indica ted that the apoptotic rate was significantly increased after HMME PDT [(34.82 ± I.42) ％ vs (3.12±0.28) ％,P＜0.05],and apoptotic rate was higher than necrosis rate [(14.65±1.02) ％ vs (34.82±1.42) ％,P＜0.05].Conclusions Low level exposure to 630 nm PDT mediated by HMME appears to induce fibroblast apoptosis.

BACKGROUND: The existing electrocardiogram(ECG)measurement strongly depends on medical professionals and inefficient high-intensity,or relies on automatic identification method which is not accurately enough.Thus,this is difficult to meet high-speed testing,accurate results and ease application for common people.OBJECTIVE: To develop a new method that was simple and efficient to apply and very easy to learn.METHODS: Algorithms were programmed and test software was developed by delphi7.0.ECG was drawn on screen.The apex,the starting point and the ending point as well as the J-point of each ECG wave were clicked by mouse or stylus.Then the wave parameters and an initial diagnosis could be quickly obtained by test software.RESULTS AND CONCLUSION: The parameters of ECG waveform such as wave height,wave time,PR interval,ST segment,QT segment,PP/RR time,cardiac electrical axis and so on could be accurately measured,and heart rate,heart rhythm and the deflection of cardiac electrical axis could be diagnosed correctly.The method was simple to learn and easy to imply,and it was also efficient,quick and accurate.Thus,it could greatly improve the efficiency of measurement and analysis for specialists,and could meet application requirements of general medicals and ordinary people.

Bronchial Diseases ; diagnosis ; diagnostic imaging ; physiopathology ; therapy ; Bronchioles ; pathology ; physiopathology ; Child ; Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Infant ; Male ; Respiratory Function Tests ; Retrospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

Objective To explore the effect of lysophosphatidic acid(LPA)on blood-brain barrier(BBB) permeability and its possible mechanism.Methods LPA or LPA+suramin(L+S)were stereotaxically injected into the right eaudate nucleus in SD rats in vivo.Evans blue(EB)was used to quantitatively measure the permeability of BBB at different time points.The expression of matrix metalloproteinase-9 was detected by immunohistochemistry technique.The pathological ultrastruetural changes of BBB were assessed by transmission electron microscopy.Results The BBB permeability began to increase after LPA administered into ipsilateral eaudate nucleus,and reached the peak at 24h.Then the permeability of BBB gradually lowered after 48h.In comparison with the same time points of control group,there were quite significant differences(P＜0.01).After L+S was injected,the change of BBB permeability had differences in comparison with those of LPA group in the same time points,(P＜0.05).MMP-9 positive cells were mainly vascular endothelial cells.The numbers of MMP-9 positive blood vessels grew at 6h in LPA group,and the expression of it reached maximum at 24h,then the number of it decreased at 48h,showing significant statistical differences in comparison with the L+S group(P＜0.01),It was observed microscopically that ultrastrueture of BBB of the LPA group was changed sharply,such as basement membrane roughed and fragmented,astroeyte end-feet swolled markedly and perivaseular space enlarged obviously.But there were no remarkable changes in BBB in L+S group.Conclusion LPA can induce increase of BBB permeability and its possible mechanism is the strong expression of MMP-9 protein produeted by endothelial cells through the mediation of LPA receptor,leading to degradation of basement membrane.

Objective To explore an ideal approach to establish rabbit model of hepatic fibrosis suitable for radiological and serological research.Methods Totally 40 rabbit models of hepatic fibrosis was established by intraperitoneally injecting 5% carbon tetrachloride in oil solution or 100% carbon tetrachloride(both groups,0.1 ml/kg,once a week),and drinking 5% ethanol water.Another 8 rabbits who drank normal water and received a peritoneal injection at 0.1 ml/kg served as control.The rabbits(5 or 2 per time for model group and control) were killed at 6,8,10 and 12 weeks respectively after first injection.Their livers were resected for gross and morphological observation with HE and Masson staining.Results Death of the models usually happened within 4 weeks after first injection and became stable after 6 weeks.Mortality of 5% carbon tetrachloride group was 60%,but 25% in 100% carbon tetrachloride group.Macroscopy and microscopy indicated that liver fibrosis was observed in rat models.Conclusion Long-term intraperitoneal injection of carbon tetrachloride results in rabbit hepatic fibrosis.Injection of 100% carbon tetrachloride intraperitoneal,at 0.1 ml/kg,once a week,combined with and 5% ethanol as drinking water is a suitable approach to establish rabbit model of hepatic fibrosis with low mortality and high success rate.

AIM: To investigate the characterization of absorption of hematoporphyrin monomerthyl ether (HMME), a domestic new generation photosensitizer product, by activated T cells from human peripheral blood. METHODS: Evaluation was performed by flow cytometry on the effects of incubating concentration and time of HMME on absorption by activated T cells. Lymphocytes were separated from human peripheral blood by density gradient centrifugation with Ficoll and T cells were activated with polyclonal stimulators PHA and PDB+Ion. To analyze the effects of HMME incubating doses on the absorption of activated T cells, the cultural lymphocytes were incubated with a serial doses of HMME for 1 h and HMME absorption were measured by FACS after immuno-staining with anti-CD3 antibody. To test the impact of HMME incubating time on the absorption of activated T cells, the cultural lymphocytes were incubated with HMME for various times and HMME absorption were measured by FACS after immuno-staining with anti-CD3 antibody. RESULTS: The HMME absorption-dose curve and absorption-time curve were shifted to right and up in the activated T cells as compared to resting T cells. HMME absorptions of activated T cells were statistic significantly larger than that of resting T cells in the doses between 5 mg/L to 20 mg/L. HMME absorptions of either activated T cells or resting T cells underwent a gradual increase with the incubation-time in HMME at concentration of 10 mg/L. HMME absorptions of activated T cells were statistic significantly larger than that of resting T cells in the incubation-time between 15 to 60 min. CONCLUSION: The differences of HMME absorption between activated T cells and resting T cells depend on the incubation times and doses of HMME. HMME absorption of activated T cells are significantly larger than that of resting T cells in certain incubation-times and doses. These results suggest that incubation time and dose associated with HMME-PDT therapeutic windows will be created for selective deletion of activated T cells.

Objective To detect the expression of human telomerase reverse transcriptase (hTERT) mRNA in the melanoma, and to analyze the relationship between the expression and subtypes and clinicopathological features of melanoma. Methods Expression of hTERT mRNA was detected by real-time quantitative PCR in 64 cases of melanoma and 30 cases of nevus. SPSS 17.0 software was used to analyze the relationship between hTERT mRNA expression and clinical pathological features of melanoma. Results The relative expression of hTERT mRNA in melanoma tissues was higher than that in nevus tissues [(52.43±5.42) vs (21.38±3.73), t= 4.72, P= 0.000]. The expression of hTERT mRNA in melanoma had no significant correlation with age, gender, ethnicity (all P> 0.05), but had relationship with subtypes, lymph node metastasis, Clark classification (all P< 0.05). The expression of hTERT mRNA in mucosal melanoma was significantly higher than that of acral and non-acral melanoma (t= 7.71, P= 0.001), while the expression of acral and non-acral melanoma had no difference (P> 0.05). Conclusions The expression of hTERT mRNA in melanoma is high, especially in mucosal melanoma. hTERT may play an important role in the occurrence and development of melanoma.