Objective To evaluate the endotoxin-neutralizing activity of modeling peptides from the limulus antilipopolysaccharide factor (MPLALFs, Ms) in vitro. Methods The endotoxin-binding activity of Ms was examined by biosensor technique and shown in values of Kon and Kd. The endotoxin-neutralizing effect was analyzed by limulus amebocyte lysate test. Results The biosensor technique results showed that the Kon values of M_0, M_1, M_2, M_3, M_4, M_6, M_8 and M_ 10 binding to LPS 055∶B5 were (840?5.716), (549?6.532), (842?6.530), (627?2.450), (996?5.716), (814?8.982), (556?1.633) and (635?2.449) arc second, of which M_4 and M_1 had the highest and lowest endotoxin-binding activity, respectively. The M_4 reacted to LPS with a Kd of 72.377 ?mol/L. The results obtained by the limulus amebocyte lysate test were the same with those from the biosensor technique. Conclusion M_4 has a potential good endotoxin-neutralizing effect in vitro.

In order to construct a single chain fragment variable (ScFv) phage display library against ovarian tumor, by using RT-PCR, the human heavy chain variable region genes (VH) and light chain variable region genes (VL) were amplified from lymphocytes of ovarian tumor patients and subsequently assembled into ScFv genes by SOE. The resulting ScFv genes were electrotransformed into E. coli TG1 and amplified with the co-infection of helper phage M13KO7 to obtain phage display library. The capacity and titer of the resulting library were detected. The phage antibody library with a capacity of approximately 3 x 10(9) cfu/microg was obtained. After amplification with helper phage, the titer of antibody library reached 5 x 10(12) cfu/mL. Human ScFv library against ovarian tumor was constructed successfully, which laid a foundation for the screening of ovarian tumor specific ScFv for the radioimmunoimaging diagnosis of ovarian tumor.

Objective To investigate the clinical characteristics of acute inhalational metallic nickel poisoning and its treatment effect.Methods Data including epidemiology,clinical features,laboratory tests,imaging examinations,therapy protocol as well as prognosis were collected and analyzed.Results Thoracic CT scan of all patients demonstrated as bilateral diffuse infiltration.Nickel concentration in both blood and urine were high in all patients and the concentration was positively associated with poisoning severity.All 4 patients received glucocorticoid treatment,2 of whom with acute respiratory distress syndrome were admitted to ICU to initiate mechanical ventilation and were given sodium dimercaptopropanesulfonate.All patients survived to hospital discharge.Conclusions Acute inhalational metallic nickel poisoning is uncommon and early symptoms are inconclusive.The blood and urine nickel concentration is helpful to judge the severity of the disease.Pulmonary diffuse infiltration is the main clinical feature.Glucocorticoid,chelation therapy as well as symptomatic treatment are effective therapies to relief disease severity.

Summary: Receptor mediated gene delivery is a new gene transfer strategy. Asialoglycoprotein receptor (ASGP-R), the receptor of asialoorosomucoid (Asor), is specially expressed on the surface of hepatocyte. In this paper, the nuclide 131I was combined with Asor to form a kind of soluble nuclide-protein complex, which can be specifically endocytosed into hepatocyte by ASGP-R. After intravenous injection of the complex into experimental animals, the deposition of Asor in vivo and the targeting quality of hepatocyte was detected by ECT. This research testified the feasibility of targeting Asor complex delivery to hepatocyte mediated by ASGP-R in vivo, and provided foundation for the genetic diagnosis and gene therapy of hepatic cell-related diseases.

Objective To investigate the expression of T cell immunoglobulin-and mucin-domaincontaining molecule-3 (Tim-3) in peripheral CD8 +T cells and its significance in patients with chronic hepatitis B (CHB).Methods Fifty-eight CHB patients and 16 healthy controls were enrolled.Tim-3 expression in CDs + T cells was detected by flow cytometry,and quantities of IFNγ-producing HBV-specific cytotoxic T lymphocytes (CTLs) in HLA-A2 positive subjects were detected by enzyme-linked immunosorbent spot (ELISPOT) test before and after the blockade of Tim-3/Tim-3L pathway.Paired t test was performed to compare the quantities of CTLs before and after the blockade,and nonparametric Spearman correlation analysis was performed to explore the correlation in quantitive data.Results Tim-3 expression in CHB patients was (14.2 ± 8.98 )％,which was higher than that of healthy controls (4.80 ± 2.92)％,and the difference was of statistical significance (x2 =92.48,P ＜ 0.05 ) Tim-3 expressions in 16 severe CHB patients and 42 mild CHB patients were ( 19.54 ± 10.95) ％ and (9.58 ± 7.30) ％,respectively,and the difference was statistically significant (x2 =77.24,P ＜ 0.05 ). Before the blockade of Tim-3/Tim-3L pathway,IFNγ-producing HBV-specific CTLs were 7.27 ± 3.14,and it increased to 19.62 ± 4.97 after the blockade ( t =2.95,P ＜ 0.05 ).Conclusion The upregulation of Tim-3 on peripheral CD8 + T cells may inhibit HBV-specific CTLs,and the blockade of Tim-3 pathway can enhance the proliferation of IFNγ-producing HBV-specific CTLs,thus can enhance antiviral effect.

Objective High-throughout sequencing of all plasmid of 206 strains of Klebsiella pneu-moniae using Solexa/Illumina sequencing technology in order to investigate the resistance for plasmids in Klebsiella pneumoniae. Methods Bacterial isolates were obtained over the years 2002-2008. Solexa/Illumi-na sequencing technology was used to sequence both samples (S1 and S2) to a depth of between 10-560 fold coverage. We used SOAP provided by BGI to find SNPS and use velvet package to assemble these sequences and gained some long sequences, and MAQ programs developed in the laboratory were used to annotate SNPs and compare lineage-specific mutations in SHV-ESBLs. Results The Metagenome of plasmid encodes a 13 variety of resistance-related genes with exceptionally high copy numbers, including ABC-type efflux pumps and 4 variety of β-lactamases, SHV-ESBLs is abroad presence. We systematically investigated single nucleo-tide substitutions in plasmids metagenome, and found an amount of nonsynonymous mutations in the SHV-ESBLs genes. Conclusion Probabily in press of positive selection, we can clearly see these nonsynonymous changes predominantly occurred in plasmid SHV-ESBLs genes. And our findings indicate a unspecial low-level resistance contribute to antimicrobial efflux in the metagenome of plasmid in Klebsiella pneumoniae.

ObjectiveTo investigate the early clinical risk factors of severe acute inhaled organic fluorine poisoning.Methods The clinical data of patients with acute poisoning of organic fluorine inhalation admitted since 2004 in Northern Jiangsu People's Hospital were retrospectively analyzed. According toDiagnostic Criteria of Occupational Acute Fluorohydrocarbon Poisoning(GBZ66-2002), all the patients were divided into three groups: mild, moderate and severe poisoning groups, the severe cases were included in the intensive group, and the others were grouped in the non-intensive group. The contents in the survey were as follows: gender, age, vital signs on admission (body temperature, pulse rate, respiratory rate, systolic blood pressure), arterial blood gas analysis record〔arterial oxygen saturation(SaO2), oxygenation index(PaO2/FiO2), lactic acid(Lac) and arterial partial pressure of carbon dioxide(PaCO2), pH value(pH)〕. Before treatment, the white blood cell(WBC) count, platelet(PLT) count, levels of alanine transaminase(ALT), creatinine(Cr), blood glucose, electrolytes(potassium, sodium, chloride, calcium), creatine kinase isoenzyme(CK-MB), etc. were examined and recorded. All the patients were immediately arranged for bedside chest X-ray examination, and the chest X-ray lung injury scores were recorded. By univariate and multivariate logistic regression analyses, the receiver operating characteristic curve(ROC curve) was drawn to evaluate the diagnostic value of the clinical risk factors.Results Sixty-two cases consisting with the standard criteria of enrollment were collected in the study, 36 cases being in intensive group and 26 cases in non-intensive group. The univariate analysis showed that the differences in pulse rate, respiratory rate, PaO2/FiO2, WBC, SaO2, Lac, pH, and lung injury score were statistically significant(P<0.05 orP<0.01). Logistic multiple regression analysis showed that PaO2/FiO2, WBC, Lac and chest X-ray lung injury score were the four indexes for predicting the independent risk factors of severe acute inhaled organic fluorine poisoning. The area under ROC curve(AUC) of PaO2/FiO2 was the highest(0.884), 95% confidence interval(95%CI) was 0.784 - 0.984, the critical value was 96.5 mmHg(1 mmHg=0.133 kPa), with the sensitivity of 75.6%, specificity of 95.2%, positive predictive value(PPV) of 92.3% and the negative predictive value(NPV) of 71.4%, in sequence, the rest were WBC(AUC 0.846, 95%CI 0.728 - 0.965, the criticalvalue 12.15×109/L), Lac(AUC 0.800, 95%CI 0.662 - 0.938, the critical value 4.2 mmol/L), chest X-ray lung injury score(AUC 0.795, 95%CI 0.652 - 0.938, the critical value 2.50), the sensitivity of the above three items was 90.2%, 83.6%, 88.5%, specificity was 90.2%, 83.6%, 88.5%, the PPV was 86.7%, 82.4%, 85.8% and NPV was 72.0%, 73.9%, 69.2% respectively.ConclusionThe blood WBC count, Lac, PaO2/FiO2 and chest X-ray lung injury score can be used as the early clinical risk factors of severe acute inhaled organic fluorine poisoning.

Objective To investigate the expression and clinical significance of platelet activating factor [PAC]-1, CD62P and TPP hi severe sepsis. Method Patients with severe sepsis who were admitted into the EICU of Subei People's Hospital from April 2007 to March 2008 were included. Patients with severe sepsis (Group Ⅲ)were treated according to the treatment guidelines for severe sepsis, and were divided, according to their clinical records, into those who survived and those who died within 28 days of admission. Patients admitted during the same period with symptoms of infection but without severe sepsis were included as the General Infected Group (Group Ⅱ). A Control Group (Group Ⅰ) comprised patients who visited the hospital over the same period for physical examination or the healthy volunteers. The group members were all included randomly, and the gender and sex of patients in all three groups were similar. Patients with acute brain infarction, acute coronary syndrome,serious diabetes, hyperlipidemia, malignant tumor, leukemia, primary liver, renal and hematopoietic system dis-eases,long-term bedridden patients, pregnant women, and patients taking hormone treatment or hranunosuppres-sants were excluded from the study. Morning venous blood was collected and ELISA and Flow Cytometry performed on the fwst day of admission for Groups Ⅰ- and Ⅱ, and on the first, third and fifth day after admission for Group Ⅲ, to determine the TpP,PAC-1 and CD62P respectively; and the Marshall score was determined. Data were ana-lyzed by SPSS 12.0 software. For continuous variables, comparisons among groups were analyzed by ANOVA.Levene's and LSD test were applied to assess homogeneity. Bivariate test is applied to Correlation Analysis. P<0.05 was regarded as a statistically significant difference. Results There were a total of 20 patients each in GroupⅠ-and GroupⅡ, and 30 in Group Ⅲ; of these, 19 were classed as survivors and 11 died during the 28-day peri-od. On the first day of admission, there were no significant differences in PAC-1, CD62P or TpP expression between Groups Ⅰ- and Ⅱ(P>0.05); however, Group Ⅲ was significantly different compared with both Group Ⅰ and Group Ⅱ (both:P<0.05). The expression of PAC-1, CD62P and TpP tended to decline in the survivor group,and became normal with the treatment process, while the expression of PAC-1 ,CD62P and TpP in the patients who died remained high, and even increased significantly over time. On the first day, the expression of CD62P and TpP in the patients who survived and in those who died was not significantly different (P>0.05); on the third day,however, a significant difference appeared with values of (2.89±1.48) % vs. (5.04±2.57) % (P<0.01) for CD62P, and (5.24±2.22) mg/L vs. (9.20±1.93) mg/L (P<0.01) for TpP. The expression of PAC-1 was significantly different between the two subgroups on the first day, with values of (3.15±0.42)% vs. (5.30±.48)% (P<0.01). The Marshall score of the two groups showed similar changes. Correlation analysis showed that PAC-1, CD62P and TpP were significantly correlated with the Marshall score. Conclusions Platelet activation and microthrombosis existing in the early stage of severe sepsis work together in the early hypercoagulable state.They both play important roles in disease development and progression. The dynamic detection of CD62P and TpP is beneficial to the diagnosis and prognosis of severe sepsis.PAC-1 appears to hold a risk stratification effect, as pa-tients with high expression of PAC-1 in the early stage show poor prognosis. Therefore, PAC-1 could be used as a marker of severe sepsis and poor prognsis.

Objective To compare the distribution situation of human papillomavirus(HPV)infective genotypes in normal cells and atypical squamous cells of undetermined significance(ASC-US)in uterine cervix and its clinical significance.Methods The pol-ymerase chain reaction(PCR)combined with the gene-chips technology were adopted to detect 23 kinds of HPV genotype from 1 000 cases of normal cells specimens and 229 cases of ASC-US specimens.Results 106 cases of HPV-positive infection were de-tected from 1 000 cases of normal cells with the total HPV infection rate of 10.60%(106/1 000),in which the single genotype in-fection rate was 9.30%(93/1 000)and the multiple genotypes infection rate was 1.30%(13/1 000);116 cases of HPV-positive in-fection were detected from 229 cases of cervial ASC-US specimens with the total HPV positive rate was 50.66% (116/229 ),in which the single genotype infection rate was 34.06%(78/229)and the multiple genotypes infection rate was 16.59%(38/229).The total HPV positive rates,single and multiple genotype infection had statistically significantly differences between the two groups(P<0.05).Conclusion The HPV types 16,18,33,42,43,52,58 are the predominant genotypes in normal cervical cells and ASC-US. PCR combined with the gene-chip technology can be used in the HPV genotype detect in cervical cells,conduces to perform the fur-ther distribution management on ASC-US and has the important significance to prevention and control of cervical cancer.

To explore the possibility to employ　99mTc-MIBI to monitor biological response of tumor cells after irradiation and to observe the relation between the radiation doses and the uptake levels of　99mTc-MIBI in tumor cells, the cells were irradiated with a single dose of 2 Gy, 10 Gy and 20 Gy respectively. The uptake of　99mTc-MIBI in each dosage group was determined before and 24, 48, 72 h after irradiation respectively. Apoptosis index (AI), plating efficiency (PE) of tumor cells was simultaneously determined. There was a positive correlation between uptake levels of　99mTc-MIBI and AI(r=－0.91, P<0.05). A negative correlation was noted between the uptake levels and PE (r=－0.86, P<0.05). It is suggested that　99mTc-MIBI may be used as a tracer to monitor the change of viability state of tumor cells after being irradiated with different doses.