Objective To evaluate the number and function of peripheral Vγ2Vδ2+T lymphocytes during Mycobacterium tuberculosis(MTB)infection in human immunodeficiency virus (HIV)infected individuals.Methods Seventy-six HIV/acquired immune deficiency syndrome(AIDS) patients co-infected with MTB were divided into active tuberculosis(TB)group(HIV+TB)and latent TB group(HIV+LTB).T cell subsets of peripheral blood lymphocytes were analysed by flow cytometry.Stimulated by protein purified derivative(PPD)and hydroxymethylbutenyl diphosphate (HMBPP),specific interferon(IFN)-γ producing T cells were detected using enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS).Data were analyzed by t test.Results The absolute number of CD3'T cells and DroDortion of V72VB2'T cells in CD3+T cells in HIV+TB group were both significantly lower than those in HIV+LTB group(t=-3.67,P<0.01;t=-2.06,P<0.05).PPD-specific IFN-y-producing T cells and percentage of PPD-specific CD4+Tcells in CD3+T cells in HIV+LTB group were both similar with those in HIV+TB group.While HMBPP-specific IFN-γ-producing T cells and percentage of HMBPP-specific Vγ2Vδ2+T cells in CD3+Tcells in HIV+LTB group were both higher than those in HIV+TB group(t=2.71 and t=3.003,respectively;both P<0.0 1).Conclusion The number and function of Vγ2Vδ2+T cells were impaired in HIV/AIDS patients coinfected with active MTB infection,which indicates that Vγ2Vδ2+T cells may be the key immune cells against MTB in individuals with impaired CD4+T cells.

Objective To investigate the incidence,bacterial pathogen and risk factors of bacterial meningitis after major craniotomy.Methods Clinical data of patients who underwent at least one craniotomy in Huashan Hospital Affiliated to Fudan University in 2008 were collected.All subjects were ≥ 18 years old,and survived at least 7 days after surgery.Patients with only cerebrospinal fluid drainage,burr holes,cranioplasty,vascular interventional surgery,transsphenoidal or spinal surgery were excluded.Risk factors for bacterial meningitis after major craniotomy were analyzed by Logistic regression.Results A total of 691 patients were enrolled,in 60 of whom (8.68％) bacterial meningitis was identified.Among 44 samples,5 were positive in culture with 2 of Acinetobacter baumannii,1 of Enterococcus faecalis,1 of Streptococcus intermedius and 1 of Klebsiella pneumonia.Diabetes (OR =5.79,95％ CI:1.40-23.93,P =0.02),Glasgow Coma Scale score ＜ 12 (OR =6.56,95％ CI:1.17-36.80,P =0.03),external ventricular drainage (OR =4.31,95％ CI:1.57-11.82,P =0.01),and continuous lumbar cistern drainage (OR =22.82,95％ CI:10.31-50.52,P =0.00) were independent risk factors for bacterial meningitis after major craniotomy.Patients with external ventricular drainage ＞ 7 d were 11.82 times more likely to develop bacterial meningitis,and those with continuous lumbar cistern drainage ＞ 10 d were 25.50 times more likely to develop bacterial meningitis.Conclusions Bacterial meningitis remains a common complication after major craniotomy,and most are induced by Gram-negative bacilli.Diabetes,Glasgow Coma Scale score,external ventricular drainage and continuous lumbar cistern drainage may increase its incidence.

Objective To evaluate the performance of peripheral blood T-SPOT.TB and cerebrospinal fluid (CSF) interferon (IFN)-γ detection in the diagnosis of tuberculous meningitis (TBM).Methods Among the 182 consecutive cases with suspected TBM in Huashan Hospital from March 2011 to March 2013,30 patients were included in the case group according to the latest diagnostic criteria of TBM.Thirty-nine patients diagnosed with non-tuberculous meningitis were included in the control group.T-SPOT.TB was employed to detect tuberculosis-specific IFN-γ-secreting T cells in the peripheral blood.And IFN-γ in CSF was detected simultaneously by enzyme-linked immunosorbent assay (ELISA) without antigen stimulation.The CSF was collected from 10 patients of TBM group after anti tuberculosis treatment for 4 weeks to observe the dynamic changes.The t-test was used for analysis of continuous variables with normal distribution and Kruskal-Wallis test was used for analysis of variables with abnormal distribution.Results ()f the 30 TBM cases,6 were confirmed cases and 24 were highly suspected cases.The control group was comprised of 12 viral encephalitis,16 suppurative meningitis and 11 cryptococcal meningitis.The positive rate of T-SPOT.TB was significantly higher in TBM group compared with control group (70％ vs 13％,x2 =12.15,P＜0.01).The mean concentration of CSF IFN γ of TBM group was 244.35 pg/mL,which was significantly higher than that of control group 9.48 pg/mL (Z=-4.646,P＜0.01).The CSF IFN-γ was significantly decreased after 4 weeks of treatment (271.02 pg/mL vs 81.36 pg/mL,Z=-3.099,P=0.002).The sensitivity and specificity of peripheral blood T-SPOT.TB in the diagnosis of TBM were 70％ and 87％,respectively.The area under the receiver operating characteristic (ROC) curve of CSF IFN-γ for TBM diagnosis was 0.819; the optimal cut-off point was 81.36 pg/mL; the corresponding sensitivity and specificity were 83 ％ and 85 ％,respectively.Conclusion Both the detection of peripheral blood T-SPOT.TB and CSF IFN-γ are of great importance for the diagnosis of TBM.Dynamic observation of CSF IFN-γ is important for disease monitoring.

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 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.

Objective To discussion the application value of three dimensional (3D) printing in neurosurgery.Methods A retrospective analysis of 7 patients with intracranial diseases,admitted to our hospital from April 2016 to December 2016 was performed.Cerebral hemorrhage,meningioma in the central region,sphenoid ridge meningioma,pituitary adenoma,odontoid deformity,chordoma,and aneurysm were noted in these patients.Cranial CT/MR imaging data of all patients were attained preoperatively.Three-dimensional reconstruction was performed using MIMICS.Models of intracranial diseases were printed using a 3D printer.Surgical simulation and planning were performed on the physical models.Results Models of intracranial diseases of all patients were produced before surgeries.The models clearly demonstrated the morphologies of intracranial diseases and spatial relationship with adjacent large vessels and skull.The operative approach was planned in vitro.Patients recovered well postoperatively without severe complications or death.Conclusion The 3D printing can assist the resection of intracranial lesions well and has great utilization potential in localizing intracranial lesions and demonstrating adjacent anatomical relationships;it provides required help to clinical doctors for preoperative planning.