Hantavirus is the main cause of hemorrhagic fever with renal syndrome (HFRS). It is an acute infectious diseases characterized by fever, hemorrhage, nephritis or thrombocytopenia, and hantavirus pulmonary syndrome(HPS). The main clinical manifestations are fever, hemorrhagic lesion, acute respiratory distress and capillary leakeage syndrome. These are four different serotypes of the hantavirus species: Hantan virus(HTNV),Seoul virus(SEOV),Dobrava/Belgrade virus(DOBV),and Puumala virus(PUUV). They are known to cause HFRS, while Sin Nombre virus(SNV) causes HPS. In China, these are two serotypes of hantavirus: HTNV and SEOV found. The severity of infection depends on the viral serotype. To find a safe, rapid and specific serotyping diagnosis of the causative virus is important. The results not only can be beneficial for rodent control, but also for prevention and therapy. The current research of Hantavirus nucleocapsid protein used as serotyping antigen are summarized.

Objective To establish the quantitative method for detecting perforin mRNA. Methods The competitive templates were prepared by restriction endonuclease method.The quantitative RT-PCR assay was established and used to detecting the perforin mRNA level in peripheral blood of tumor patients and healthy adults.Results Average perforin mRNA level of six healthy adults is 0.51±0.40 pmol/ml.The perforin mRNA level of patients with digestive tumors are significantly lower than that of healthy adults(P<0.01).But compared with healthy adults,no significant difference is showed in patients with breast cancer or leukemia.Conclusion The decreased perforin mRNA level probably attributes to the high incidence of cancers.Except for perforin-mediated cytolysis,additional effective mechanism against tumor cells maybe exist.

Objectives: To express and purify N-terminal peptide of human perforin(hPFP-N). 　Methods: The recombinant expressive plasmid pGEX-KG/hPFP-N was constructed and then introduced into a strain of E. coli BL21(DE3). Upon the induction of IPTG, GST/hPFP-N fusion protein was expressed. The expressed fusion protein was localized in inclusion bodies which could be solubilized by sonication after detergent lauroylsarcosine was added. The fusion protein was purified by affinity chromatography with glutathione agarose. After being cleaved by thrombin, GST and uncleaved fusion protein were removed by glutathione agarose beads once more, then purified recombinant rhPFP-N protein was obtained.　 Results and Conclusions: GST/hPFP-N fusion protein can be effectively expressed in E. coli and the protein hPFP-N was obtained after the purification process.

Objective To study the vulnerability of astrocytes bearing mutant SOD1 under the oxidative stress.Methods The cytotoxicity of the serum deprived astrocytes was measured by MTT.The level of ROS was shown by fluorescence of DCF through confocal microscopy.The expression of Nrf2,HO1 and NQO1 in the different cells was detected by Western blot.Results The level of cellular toxicity was higher in the astrocytes bearing mutant SOD1 exposed to the oxidative stress than the astrocytes hearing wild type SOD1.In the astrocytes bearing mutant SOD1,the expression of Nrf2,HO1 and NQO1 decreased.In the presence of mutant SOD1,an unexpected 44 per-cent decrease of Nrf2 was detected.This was associated with a decreases in multiple downstream phase Ⅱ detoxif-ying enzymes and antioxidant enzymes,known as NQO1 and HO1.Furthermore,our results showed that the ex-pression of NQO1 increased 1.5 and 2.5-fold by EGCG at 5 and 10 μmol/L.EGCG also elevated the expression of total Nrf2.Confocal microscopy showed that EGCG caused Nrf2 translocation from the cytoplasm to the nucleus.Conclusion Decrease in Nrf2 expression is the mechanism to explain the vulnerability of astrocytes bearing mutant SOD1 and EGCG strengthened antioxidation function by upregulating the activity of Nrf2.

BACKGROUND: Although angiography is the gold standard for the diagnosis of cerebrovascular disease, it cannot be used widely because of its traumatotaxis. Transcranial Doppler ultrasound (TCD) and MRA are both non-invasive examinations, which can be used to evaluate intracranial large artery vascular trunks and hemodynamics.OBJECTIVE: To investigate the reliability of diagnosis with TCD on hemodynamic changes of ischemic cerebrovascular disease (ICVD) and the relativity between TCD and MRA.DESIGN: Case analysis based on patients.SETTING: Neurological Department of the First Affiliated Clinical Hos-pital, Harbin Medical University.PARTICIPANTS: From April 2001 to February 2002,totally 45 patients with stroke,selected from outpatients and inpatients of Neurological Department of First Affiliated Clinical Hospital of Harbin Medical University,examined by CT, to be told the fact and accepted, were taken part in the study.METHODS: Patients were examined with TC-2021 TCD instrument produced by German EME Corporation and VISART1.5T superconduct MR instrument produced by Toshiba. Totally 45 patients had TCD and MRI examinations, and the alternation between two examinations was 1 to 15 days. The hemodynamics and morphological changes were observed including middle cerebral artery (MCA), end of internal carotid artery (ICA),anterior cerebral artery (ACA), posterior cerebral artery (PCA), basal artery (BA), and vertebral artery (VA).large artery vascular trunks and hemodynamic results with TCD and MRA;ty, false positive and false negative with TCD with those of MRA.TCD and MRA: Theory data was Tmin=9.91; χ2=107.92, P ＜ 0.005, it was considered that the results with the rate of TCD were related with that of MRA. The detection rate of TCD was 19.06%, and that of MRA was 15.25%. There were significant differences between the detection rate of standard, the sensibility of diagnosing abnormal vessels by TCD was 78.10%, specificity 94.22%, false positive 7.46%, false negative 23.31%,and the coincidence rate 89.36%.CONCLUSION: TCD can assess exactly the functional status of the vessels. It can also reflect early and sensitively the change of hemodynamics of cerebral blood flow. While MRA can display the change of vascular appearance directly so that the combination of TCD and MRA will be improve sensibility and specificity of diagnosis.

Objective To investigate the role of activation of Akt/mTOR pathway in denfense against UVB-induced apoptosis in cultured human skin keratinocyte cell line HaCaT. Methods HaCaT cells were irradiated with UVB at different doses for various durations. Western blotting was performed to detect dynamic changes of Akt/mTOR pathway-related signaling molecule, such as phosphorylated-epidermal growth factor receptor (EGFR), -Akt, -4EBP1, etc; apoptosis was estimated by staining with DNA dye Hoechst 33342. To evaluate the role of signaling molecules in defense against UVB-induced apoptosis, HaCaT cells were pretreated before irradiation with EGFR inhibitor (PD153035), PI3K inhibitor (LY294002), mTOR inhibitor (rapamycin) followed by the detection of expressions of signaling molecule and apoptosis. Results UVB could activate Akt/mTOR pathway in a dose- (5 ～ 30 mJ/cm2) and time- (5 ～ 30 min) dependent manner. PD153035,LY 294002 and rapamycin could inhibit UVB-induced activation of the Akt/mTOR pathway. The apoptosis rate in HaCaT cells was upregulated by pretreatment with rapamycin and LY294002. Conclusion The activation of Akt/mTOR signaling pathway could inhibit the UVB-induced apoptosis in cultured HaCaT cells.

ObjectiveTo observe the efficacy of a Levofloxacin-containing 10-day sequential therapy for Helicobacter pylori eradication.Methods Totally 86 Helicobacter pylori-infected patients with chronic gastritis or peptic ulcer diagnosed by gastroscopy were randomized into two groups: the trial group and the control group (n =43 in each group).The trial group was treated with Esomeprazole 20 mg bid plus Amoxicillin 1000 mg bid at the first 5 days,and followed by Esomeprazole 20 mg bid,Levofloxacin 500 mg qd and Furazolidone 100 mg bid,at the last 5 days;The control group was treated with standard triple regimen: Esomeprazole 20 mg,Amoxicillin 1000 mg and Clarithromycin 500 mg,all bid for 7 days.The eradication rates of Helicobacter pylori in the two groups were compared after completion of treatment.ResultsAfter treatment,the eradication rates of Helicobacter pylori were significantly different between the trial and control group(93.0％ vs 74.4％,P ＜0.05)Conclusion The 10-day sequential therapy with.Levofloxacin-containing regime can achieve a higher eradication rate of Helicobacter pylori infection.

Objective To investigate the relationship between the efficacy of autologous cultured melanocyte transplantation and serum levels of interleukin-17 (IL-17) and FoxP3 in patients with vitiligo.Methods Forty patients with stable vitiligo vulgaris were included in this study,and received autologous cultured melanocyte transplantation.Six months after the transplantation,treatment efficacy was evaluated,and patients were classified into the successfully treated group (n =25) and unsuccessfully treated group (n =15).Peripheral blood was collected from all the patients before the transplantation,and enzyme-linked immunosorbent assay (ELISA) was performed to measure the serum levels of IL-17 and FoxP3.Statistical analysis was done using two independent samples t-test with the SPSS software (version 17.0).Results The successfully treated patients showed lower serum levels of IL-17 ((15.29 ± 7.86) vs.(43.88 ± 13.02) ng/L,P ＜ 0.05),but higher serum levels of FoxP3 ((6.08 ± 2.03) vs.(3.37 ± 1.81) ng/L,P ＜ 0.05) than the unsuccessfully treated patients.Conclusion The increased serum IL-17 and decreased serum FoxP3 may contribute to the failure of autologous cultured melanocyte transplantation in patients with vitiligo.

Objective To evaluate the therapeutic efficacy of autologous melanocyte transplantation for the treatment of vitiligo in patients with abnormal thyroid function.Methods A total of 60 patients with vitiligo were enrolled in this study,including 30 with abnormal thyroid function and 30 without.Epidermal sheets were obtained by suction blister biopsy from the normal skin of all the patients followed by melanocyte isolation and culture.After 2-5 passages of subculture,the melanocytes were transplanted onto vitiliginous lesions,which were abraded previously by ultra-pulsed CO2 laser,in the corresponding patients.All the patients were followed for 6-12 months.Results Of the 30 patients with abnormal thyroid function,7 patients achieved more than 90％ repigmentation,9 patients 50％-89％ repigmentation,53.3％ more than 50％ repigmentation,with the average repigmentation rate being 47％ within 6 months after the transplantation.Meanwhile,13 out of the 30 patients without abnormal thyroid function showed more than 90％ repigmentation,11 showed 50％-89％ repigmentation,with the average repigmentation rate being 75％.Both the cure rate and response rate were significantly higher in the patients without abnormal thyroid function than in those with (cure rate,43.3％ vs.23.3％,P＜ 0.05; response rate,80％ vs.53.3％,P＜ 0.05).Significant differences were also found in the response rate for lesions on the face or neck and for those sized more than 20 cm2 between the two groups of patients (both P ＜ 0.05).The lesions transplanted with epidermal melanocytes from the waist exhibited the lowest cure rate and response rate.Conclusion Clinical or subclinical thyroid dysfunction may have a negative impact on the efficacy of autologous melanocyte transplantation in vitiligo.

Objective To determine the effect of transcranial Doppler ultrasound (TCD) combined with focal mild hypothermia on cerebral infarction treated with thrombolytic unokinase (UK).Methods32 rabbits were randomly divided into the TCD group, UK group, TCD+UK group and hypothermia group with 8 animals in each group. Mean velocities of the middle cerebral artery were recorded after unokinase infusion to estimate whether blood vessels reopened. Meantime, brain temperature of lesion site was tested to find out whether declined to hypothermia level.ResultsThe percentage of recanalization was 87.5% in the hypothermia group, 75% in the TCD+UK group, significantly higher than that in the UK group (37.5%)(P<0.05). After 30 minutes of the rabbit refrigerated at 2 ℃, the putamen temperature declined to 32 ℃～34 ℃.ConclusionApply the regional refrigerstor apparatus to decline the brain temperature to mild hypothermia level during therapeutic time window. TCD combined with focal mild hypothermia can improve the thrombolytic effect of unokinase.