This is the first report of the sensitizing effect of cation surfactant on the enhanced resonance light scattering (RLS) resulted from the interaction of organic dyes with Deoxyribonucleic acid (DNA). On the conditions of pH 10.4 ～11.8 and ionic strength being less than 0.050, the interaction of brilliant green (BG) with DNA results in enhanced RLS signals at 398.0 and 467.2 nm overloaded some shoulder peaks in the enhanced region. It was found that the interaction was sensitized by the presence of cetyltrimethylammonium bromide (CTMAB). If 2.0～3.2×10-5 mol/L CMAB was used in the interacting medium, DNA in the range of 0～1.2 (mg/L could be detected with the limit of detection being less than 5 μg/L. Four synthetic samples were analyed with reproducibility

Objective To investigate the risk factors associated with cranial nerve impairment in patients with tuberculous meningitis.Methods A total of 121 patients with tuberculous meningitis who were admitted to Huashan Hospital from 2000 to 2011 were reviewed retrospectively.Demographic data (gender,age),course of disease,initial results of cerebral spinal fluid (CSF) tests,occurrence of cranial nerve impairment and prognosis of these patients were collected.All the patients were followed up for at least 3 months,and for those with cranial nerve impairment,the minimum follow-up period was 1 year in order to judge the recovery of cranial nerve impairment.Multivariate analysis was performed to study the associated risk factors.Results Out of 121 patients,22 (18.2 ％)developed cranial nerve impairment.Nerves involved were abducens nerve,oculomotor nerve,optic nerve and auditory nerve,and impairment of single nerve occurred in 9 (40.9 ％),8 (36.4 ％),7(31.8％) and 1(4.5％) patient,respectively.Three cases had more than one group of cranial nerves involved,accounting for 13.6％ of the 22 patients with cranial nerve impairment.The incidence of conscious disturbance was significantly higher in patients with cranial nerve impairment than those without impairment (77 ％ vs 45 ％,P=0.020).Delay in diagnosis (OR =1.017,95 ％ CI:1.001-1.033,P=0.040) and occurrence of conscious disturbance (OR =3.242,95 ％ CI:1.142-9.205,P=0.027) were independent predictive factors of cranial nerve injury.During one-year follow-up,90.9％ of patients were fully recovered from cranial nerve impairment,with a median duration of 1 month (range 0.5-6.0 months).Conclusions Cranial nerve impairment is a common complication in patients with tuberculous meningitis.Delay in diagnosis and occurrence of conscious disturbance were independent predictive factors.Most cranial nerve impairment were reversible,and timely diagnosis and treatment are important ways to reduce complications.

Objective To develop a new multiplex allele-specific PCR(MAS-PCR) assay to detect mutation in codon 315 of katG gene of Mycobacterium tuberculosis , mutation in this codon has been reported to be able to account for the Mycobacterium tuberculosis clinical isolates resistant to isoniazid(INH).Method Based on the sequence of katG gene of Mycobacterium tuberculosis , three specific primers are designed to carry out the MAS-PCR, 84 purified DNA preparation with known katG 315 variation detected by PCR-restriction fragment length polymorphism (PCR-RFLP) are used to optimize PCR.Results 84 Mycobacterium tuberculosis clinical stains are detected by the MAS-PCR and PCR-RFLP, respectively.The sensitivity of detection by MAS-PCR is 77.8%,and the specificity is 95.2%.katG mutation S315N(AGC→AAC), neglected in RFLP, can be detected by MAS-PCR.Conclusion MAS-PCR assay is sensitive, specific, economic and easy to carry out , can be used in clinical laboratories to detect the INH-resistant Mycobacterium tuberculosis strains.

Objective To study the cell death in macrophages (THP-1) stimulated with different agonists (H37Rv-PPD or BCG-PPD) and to investigate the relationship between Toll like receptor (TLR)-2 and THP-1 apoptosis. Methods H37Rv-PPD and BCG-PPD were used to stimulate THP-1 cells for 3 h, 8 h, 15 h and 24 h, respectively with or without TLR-2 blockade. Cells were analyzed by flow cytometry to detect the TLR-2 expression. Annexin V staining and Hochest staining were performed to evaluate apoptosis. Results The apoptosis cells were increased when stimulated with BCG-PPD and the percentage was 30.2% at 24 h, which were confirmed by Hochest staining.However, the expression of TLR-2 did not increase simultaneously with percentage of 8.8% at 24 h.Nevertheless, most cells presented with necrosis form when stimulated with H37Rv-PPD and the expression of TLR-2 remained at high level with the percentage of 17.2% at 24 h, while the percent of apoptosis rate was only 7.7%. Under treatment of TLR-2 antibodies, the percentage of apoptosis decreased to 10.5% at 24 h of BCG-PPD stimulation and TLR-2 expressions were down-regulated to less than 3% at all time points; but after H37Rv-PPD stimulation, the percentage of apoptosis and TLR-2 expression did not changed obviously. Conclusions The attenuated BCG-PPD induces THP-1 apoptosis predominately, which is partially correlated with TLR-2 expression. While virulent H37Rv-PPD induces THP-1 necrosis predominately.

Objective To analyse the clinical feature of infective endocarditis (IE) in recent years.Methods Clinical profiles including age of onset,predisposing factor,clinical manifestation,blood culture and ultrasonic cardio gram (UCG) of 97 cases from Huashan Hospital in the recent 10 years were collected and analyzed retrospectively. Descriptive data were represented as mean ±standard deviation form.Positive rate was represented as percentage form.Fisher's exact test were used to determine two groups' comparison.Results The mean age of the population was (49±17)years.Seventy-three patients (75.3％) had background heart disease,the top 3 of which was rheumatic heart disease in 27 patients (27.8％),congenital heart disease in 23 patients (23.7％) and idiopathic mitral valve prolapse in 18 patients (18.6 ％).The most common clinical manifestation were fever (99.0％),murmurs (95.9％) and anemia (84.5％).Sixty-six patients (68.0％) had positive result of blood cultures. Streptococcus viridans,which was found in 28 patients with native valve endocarditis (42.4 ％),was still the most common pathogen.Staphylococcus,which was found in 18patients (27.3％),had an elevated ratio.Staphylococcus aureus was found in 10 patients (15.2％)and 3 of which were MRSA.Coagulase-negative staphylococcus was found in 8 patients (12.1 ％ ) and 2 of which were MRCNS. Drug-resistant bacteria was increased and pathogens were varied.Vegetations were found in 79 patients (81.4％) by UCG.ConclusionsClinical manifestation,predisposing factor and pathogen have changed in IE patients. Attaching importance to physical examination,multiple-time blood culture and UCG helps the diagnose of IE.

Objective To investigate the expression of elongation of very long-chain fatty acids family member 6 (ELOVL6) in high-grade serous ovarian carcinoma (HSOC), and explore the correlation between its expression and clinical prognosis in these patients. Methods The expression of ELOVL6 at mRNA and protein levels were respectively detected by reverse transcription (RT)-PCR and immune histochemistry method in 12 cases with normal ovarian tissues and 172 cases with HSOC from primary tumor site, forty of which had paired peritoneal metastatic tissues. Results (1) The results tested by RT-PCR showed that ELOVL6 expression in normal ovarian tissue was 4.8±1.1, while 1.2±0.7 in primary tumors and 1.8 ± 0.9 peritoneal metastatic sites in HSOC. Compared with normal ovarian tissue, the level of ELOVL6 mRNA was significantly lower in HSOC (P<0.05). There was no significant difference between primary and peritoneal metastatic sites in HSOC (P=0.610). It was shown that ELOVL6 protein localized in cytoplasm of ovarian cancer cell by immunostaining assay. (2) ELOVL6 expression was observed in all normal ovarian tissue, 70.2%of G1-G2 and 48.8%of G3 HSOC (P<0.05). ELOVL6 expression in drug-resistant group were significantly lower than that in non-resistant group (39.1% vs 65.0%, P<0.01). The median disease-free survival was 41 months in the ELOVL6-positive group and 39 months in ELOVL6-negative group (P>0.05). The total median survival was 52 months in ELOVL6-positive group and 44 months in ELOVL6-negative group (P>0.05). Conclusion Low expression of ELOVL6 may correlate with the poor differentiation and drug resistance in HSOC.

Objective To investigate the clinical features, diagnosis, treatment and prognosis of muhisystemic invasive fungal diseases. Methods Twenty-one patients with multisystemic invasive fungal diseases who were hospitalized in department of infectious diseases from January 2001 to June 2008 were retrospectively reviewed. The pathogenic bacteria, involved organs, underlying diseases, clinical manifestations, treatments and prognoses of muhisystemic invasive fungal diseases were analyzed. Results Among 21 recruited cases, 17 had underlying diseases and 11 were treated with long-term immunosuppressive agents. The main pathogenic bacteria were Cryptococcus neoformans, Aspergillus and Candida parapsilosis. Lung and brain were involved in 16 cases (skin involve in 2 cases and lymph node involved in 1 case simultaneously), lung and lumbar involved in 2 cases, heart valves involved in 2 cases, and liver, spleen and bone marrow involved in 1 case. Eight cases were cured, 6 were improved and 7 died. Conclusions In this study, most of the 21 cases with multisystemic invasive fungal diseases are immunocompromised. The main pathogenic bacterium is Cryptococcus neoformans. The lung and brain are common organs involved. Prognosis is associated with early diagnosis and active anti-fungal treatment.

Objective To evaluate the application of enzyme-linked immunospot assay (T SPOT) in diagnosis of Mycobacterium tuberculosis (MTB) infection for AIDS patients. Methods The rapid T SPOT assay was employed to detect ESAT-6 and CFP-10 specific T cells in blood samples from 35 AIDS patients with active tuberculosis. The results were compared with those of PPD skin test. Results The positive-MTB rate with T SPOT assay in all patients (n = 35 ) was 65.71%, that in patients with CD4 < 200/μ (n = 21) was 66.67%, in patients with CD4 > 200/μl (n = 10) was 60.0% , in patients with anti-tuberculosis treatment < 2 months ( n = 15 ) was 83.33% and in patients with treatment > 2 months ( n = 6 ) was 25.00%. The positive rates of PPD skin test were 24.24%, 25.00%, 33.33%, 9.09% and 25.00% in these groups, respectively. The positive rates of T SPOT assay were significantly higher than those of PPD skin test in all patients, in those with CD4 < 200/μl and with anti-tuberculosis treatment < 2 months ( P < 0.05 ). Conclusion T SPOT assay was more sensitive than conventional PPD skin test, and can be applied in the diagnosis of MTB infections for AIDS patients.

ObjectiveTo investigate the incidence of bacterial meningitis in patients who underwent intracranial neurosurgery and to explore the pathogen distribution and related risk factors.MethodsThe patients were selected by cluster systematic sampling method from the patient population who underwent intracranial neurosurgery for at least one time in Huashan Hospital affiliated to Fudan University in 2008.Chi-square test and Logistic regression model were used to analyze the data.ResultsA total of 1165 patients were included in this study.Seventy-nine of them were diagnosed with bacterial meningitis,while eight of them bad positive results for bacterial culture,including 3 cases of Acinetobacter baumannii,and 1 case each of Acinetobacter lwoffii,Klebsiella pneumoniae,Staphylococcus epidermidis,Enterococcus and Streptococcus intermedius. The common neurosurgical diseases with high incidence of postoperative bacterial meningitis were neurilemmoma 15.85％(13/82),glioma 12.21％ (21/172) and hydrocephalus 10.34％ (3/29).Logistic regression analysis identified male,implant,enteral nutrition,external ventricular drainage and wound negative pressure drainage as independent risk factors for postoperative bacterial meningitis.ConclusionsPostoperative central nervous system infection is the most common and severe complication after the intracranial neurosurgery.The results highlight the importance of improving pathogen detection rate and enhancing the prevention in high risk patient populations.

Objective To study the different response in macrophages treated with different agoβ and IL-10 in Mycobacterium tuberculosisnists(H37Rv-PPD and BCG-PPD)related with Mycobacterium tuberculosis and the relationship with TNF-αt,IL-1β and IL-10.Methods Using H37Rv-PPD and BCG-PPD to stimulate THP-1 cell for 3h,8h,15h,24h respectively.Cells were ananlyzed by Hochest staining under fluorescence microscopy to assay cell death(apoptosis and necrosis).At each stimulating time,TNF-α,IL-1β and IL-10 were examined by ELISA.Results Under fluorescence microscopy,it could easily see oval apoptotic bodies of THP-1 stimulated by BCG-PPD.However ,the nucleus were often isolated and necrosis-like when cells were stimulated by H37Rv-PPD.In a word ,BCG-PPD tend to induce THP-1 cells to apoptosis,but H37Rv-PPD inclined to induce cells to undergo necrosis.In supernatant of cells stimulated by BCG-PPD,the expression of TNF-αand IL-10 were lower than the cells stimulated by H37Rv-PPD,but the expression of IL-1β was higher than the latter.Conclusion It indicated that the necrosis of cells stimulated by H37Rv-PPD was asossiated with the excessive expression of TNF-α and IL-10,and the apoptosis of cells induced by BCG-PPD was IL-1β related.Perhaps the mechanism of differences in virulence exist in protein of strain,and associated with cytokines IL-1β,TNF-α and IL-10.