All Gene Expression of Leukotriene A4 Hydroxylase (LTA4H ) Gene in Extra Pulmonary Tuberculosis Patients and Healthy Control in North Indian Population

Initial diagnosis and timely treatment of Extra Pulmonary Tuberculosis (EPTB) continues to be a challenge in all over World as well as India. First time, this analysis will discover the role of LTA4H gene and may be establishing another candidate that impacts the sensitivity to EPTB in the population of North India. This study will be the first report on LTA4H gene various diagnostic markers, expression of gene may validate as a prognostic factor in (EPTB). The diagnosis (EPTB) poses a special challenge, as it is often missed or misdiagnosed due to its atypical presentations and difficult to isolate M tuberculosis (MTB) due to the small number of organisms present at these sites. Subsequently the outcome of present study will reinforce possible use of LTA4H as biomarkers and the therapeutic utility for (EPTB). This study will be a step to decrease the analytical and therapeutic window to identify another risk factor LTA4H for EPTB. Leukotriene A4 hydroxylase (LTA4H), an enzyme which changes LTA 4 to LTB4, controls the balance amongst the anti-inflammatory lipoxins and pro-inflammatory LTB4, with directly consequences in TB-driven inflammation. In humans and will spawn new ways to protection and enhance the wellbeing status of individuals and population groups. On RT-PCR, Extra Pulmonary Patients had lower expression of LTA4H compared to the controls. Correlation of biomarkers will reveal LTA4H level correlated with age, Gender Smoking, Clinical Parameter Serum Total Protein, BMI Height and TLC, Laboratory Parameter. On ELISA kit and follow as per manufacturer protocol. CEA562Ge 96 Tests Enzyme-linked Immunosorbent Assay Kit For Leukotriene B4 (LTB4) LTB4 Protein level in Extra Pulmonary Patients, (EPTB) (2304.52pg/ml) had lower expression of gene LTB4 compared to the controls (3096.142pg/mls) (P value = 0.0012).

Role of TRAF6 in inflammatory responses of human osteoblast-like cells with Enterococcusfaecalis / 口腔疾病防治

Objective @#Explore the role and status of tumor necrosis factor receptor-associated factor 6 (TRAF6) in the inflammatory response of human osteoblast-like cells MG63 which was triggered by Enterococcus faecalis (E. faecalis) and its lipoteichoic acid (LTA)@*Methods@# SiRNA technology was applied to silence the TRAF6 gene of MG63 cells, Using E.faecelis and its LTA to stimulate the silence MG63 cells with different hours. After that, using real-time PCR technology to detect toll-like receptor 2 (TLR2) and TRAF6 gene expression and using ELISA assay to detect proinflammatory cytokines interleukin-1β and TNF-alpha expression levels.@*Results@#When MG63 cells was infected by E. faecalis, its LTA, TLR2 and TRAF6 gene level has increased to varying degrees (P< 0.05); interleukin-1β and TNF-alpha expression was significantly higher (P< 0.05). When TRAF6 gene of MG63 cells was silenced by siRNA, pro-inflammatory cytokines interleukin-1β, interleukin-6, interleukin -8 and TNF-alpha expression decreased significantly (P< 0.05).@*Conclusion@#E. faecalis and its toxic components is identified by MG63 cells mainly through TLR2 receptors. The major virulence factor in periapical infections caused by E. faecalis is LTA.

Correlation Between the CYP2C19 Phenotype Status and the Results of Three Different Platelet Function Tests in Cardiovascular Disease Patients Receiving Antiplatelet Therapy: An Emphasis on Newly Introduced Platelet Function Analyzer-200 P2Y Test

BACKGROUND: An association has been reported between CYP2C19 polymorphism and the altered antiplatelet activity of clopidogrel. We investigated this association using the newly introduced platelet function analyzer (PFA)-200 (INNOVANCE PFA-200 System; Siemens Healthcare, Germany) P2Y test. METHODS: Polymorphisms of CYP2C19*2, *3, *17 and the degree of inhibition of platelet function were determined in 83 patients. Three different platelet function tests were used to evaluate the degree of platelet inhibition and to check the association with genotype. RESULTS: The post-procedure PFA-200 values of extensive metabolizers (EM) patients (285.3+/-38.8) were higher than those of intermediate metabolizers (IM) and poor metabolizers (PM) patients (227.7+/-98.3 and 133.7+/-99.2, respectively; P=0.024). Light transmittance aggregometry (LTA) and the VerifyNow system showed that the post-procedure values for EM patients were lower than those of IM and PM patients (LTA: 24.4+/-15.7, 34.1+/-17.6, and 42.2+/-16.9, respectively, P<0.001; VerifyNow: 133.2+/-60.5, 171.5+/-42.6, and 218.7+/-59.3, respectively, P<0.001). The high residual platelet reactivity (HPR) rates were significantly different among the EM, IM, and PM groups using PFA-200 (PM:IM:EM=82.4:40.6:11.8, P<0.001). CONCLUSIONS: Approximately, 59.0% of Korean patients with cardiovascular disease receiving clopidogrel had CYP2C19 loss-of-function genotypes classified as IM or PM, and the frequency was similar to the data from Asian people. The PFA-200, LTA, and VerifyNow platelet function tests revealed evidence of a significant association between the efficacy of clopidogrel and CYP2C19 genotypes.

Anti-tumor effect of LTA combined with 5-FU on H22 tumor bearing mice

OBJECTIVE@#To study the effect of lipoteichoic acid (LTA) and 5-FU on the expression of caspase-3, EGFR, TGF-α proteins of tumor tissue of H22 cancer bearing mice and its anti-tumor mechanism.@*METHODS@#A total of 40 SPF grade Kunming mice were selected to establish H22 liver cancer model, and then the mice were divided into 4 groups at random with ten mice in each group. Group A was given saline lavage treatment, Group B was treated with 5-FU by intraperitoneal injection, Group C was treated with LTA by lump body injection; Group D was treated with LTA by lump body injection and 5-FU by intraperitoneal injection. Two weeks after the treatment, the mice in each group were executed and the tumor tissue was stripping and weighted, and the tumor growth inhibition ratio was calculated. Then the tumor tissue was processed for conventional embedding, sectioned to observe the expression of caspase-3, EGFR, TGF-α by immunohistochemical staining method.@*RESULTS@#The tumor inhibitory rate o f Group D was significantly higher than Groups B and C (P  0.05). The IDO values of TGF-α, EGFR proteins in Groups B, C, D mice tumor tissue were significantly lower than that in group A (P < 0.05); while IDO value of caspase-3 in Groups B, C, D group mice tumor tissue was significantly higher than that in Group A (P < 0.05). The IDO value of TGF-α, EGFR in Group D mice tumor tissue were significantly lower than that in Groups B and C; While IDO value of aspase-3 in Group D was significantly higher than that in Groups B and C (P < 0.05).@*CONCLUSIONS@#LTA combined with 5-FU can effectively inhibit the tumorigenesis of H22 tumor bearing mice, increase the caspase-3 protein expression, inhibit TGF-α and EGFR protein expression, further promote tumor cell apoptosis and play a synergistic antitumor effect.

Anti-tumor effect of LTA combined with 5-FU on H22 tumor bearing mice

Objective: To study the effect of lipoteichoic acid (LTA) and 5-FU on the expression of caspase-3, EGFR, TGF-α proteins of tumor tissue of H22 cancer bearing mice and its anti-tumor mechanism. Methods: A total of 40 SPF grade Kunming mice were selected to establish H22 liver cancer model, and then the mice were divided into 4 groups at random with ten mice in each group. Group A was given saline lavage treatment, Group B was treated with 5-FU by intraperitoneal injection, Group C was treated with LTA by lump body injection; Group D was treated with LTA by lump body injection and 5-FU by intraperitoneal injection. Two weeks after the treatment, the mice in each group were executed and the tumor tissue was stripping and weighted, and the tumor growth inhibition ratio was calculated. Then the tumor tissue was processed for conventional embedding, sectioned to observe the expression of caspase-3, EGFR, TGF-α by immunohistochemical staining method. Results: The tumor inhibitory rate o f Group D was significantly higher than Groups B and C (P 0.05). The IDO values of TGF-α, EGFR proteins in Groups B, C, D mice tumor tissue were significantly lower than that in group A (P < 0.05); while IDO value of caspase-3 in Groups B, C, D group mice tumor tissue was significantly higher than that in Group A (P < 0.05). The IDO value of TGF-α, EGFR in Group D mice tumor tissue were significantly lower than that in Groups B and C; While IDO value of aspase-3 in Group D was significantly higher than that in Groups B and C (P < 0.05). Conclusions: LTA combined with 5-FU can effectively inhibit the tumorigenesis of H22 tumor bearing mice, increase the caspase-3 protein expression, inhibit TGF-α and EGFR protein expression, further promote tumor cell apoptosis and play a synergistic antitumor effect.

Investigation of the Antiplatelet Function of Clopidogrel by Thrombelastography and Light Transmission Aggregometry / 现代检验医学杂志

Objective To detect the antiplatelet function of clopidogel by Thrombelastography (TEG)and light transmmi-tance aggregometry (LTA)and analyze efficiency of two methods.Methods ①Venous blood samples were taken from 48 outparients and inpatient with acute corotary syndrome (ACS)in PLA General Hospital,including unstable anqina,ST seg-ment elevation myocardial infarction and non ST segment elevation myocardial infarction (32 males,16 females)by random number table from March to July 2011,whose average age was 73 (62~90).②All of them were served 160 mg aspirin and 300 mg clopidogrel after they were in hospital at first time,and then served with 75 mg/d clopidogrel for 9 months.After that,venous blood samples were drew from them who were served clopidogrel 2 hours later.③Platelet aggregation function was assayed with LTA and TEG.All volunteers were signed informed consent and the experiment was approved by the hos-pital ethics committee.Results The platelet aggregation rate detected with LTA induced by adenosine diphosphate (ADPL-TA-INDU)was 40~80% in 41 individuals (85.4%),>80% in 7 individuals (14.6%),coefficient of variation (CV)was 0.19.The inhibition ration of platelet function (INHI)detected with TEG (ADPTEG-INHI)was 10~60% in 40 individu-als (83.3%),60 in 6 individuals (12.5%),CV was 0.54.The sum of ADPLTA-INDU and ADPTEG-INHI (ADPLTA-INDU+ADPTEG-INHI)tended to a constant (Mean=98%,SD=12.1).There was no significant indifference between every (ADPLTA-INDU+ADPTEG-INHI)and their Mean (98%)by paired t-test.The ADPLTA-INDU was negative correlated with ADPTEG-INHI (r=-0.75,P<0.01).Conclusion The effect of clopidogrel was mild and the efficiency of TEG and LTA was similar in detecting the antiplatelet function of clopidogrel,but the preci-sion of TEG was lower than LTA.

Association Study of Single-Nucleotide Polymorphism in Lymphotoxin Alpha Gene and Bipolar I Disorder

OBJECTIVES: Proinflammatory process has been implicated as an underlying mechanism of bipolar disorder and schizophrenia. Previous studies have suggested a possible role of lymphotoxin alpha (LTA) gene in the development of schizophrenia and have prompted further investigation in bipolar patients. Association of the LTA +252A/G polymorphism with susceptibility to bipolar I disorder itself as well as with vulnerability among a subset of psychotic bipolar patients were tested. METHODS: DNA extraction was done by a standard method and genotyping was carried out by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 114 Korean patients with bipolar I disorder and 202 healthy controls. SPSS v18.0 was used for statistical analysis. Comparisons of the genotype and allele distributions in LTA +252A/G polymorphism were made using a chi-square test. The genotype and allele associations were also evaluated using odds ratio (OR) and 95% confidence interval (CI). Statistical significance was accepted when p was < 0.05. RESULTS: No significant association was found between the LTA +252A/G polymorphism and bipolar disorder. However, LTA +252G allele was present with significantly higher frequency among bipolar patients with psychotic features compared to those without (chi2 = 4.69, p = 0.034, OR = 2.495, 95% CI = 1.069-5.827). CONCLUSION: The results suggest that the allele LTA +252G of the polymorphism may be associated with the psychotic subset of bipolar disorder but not with bipolar I disorder itself. Adequately powered subsequent studies should be conducted.

Influence of moxifloxacin on lipoteichoic acid induced apoptosis and expression of inflammatory cytokines in human alveolar macrophage / 中华微生物学和免疫学杂志

Objective To investigate the lipoteichoic acid(LTA) induced apoptosis and the expression of inflammatory cytokines in human alveolar macrophage (AM) and the anti-apoptotic and anti-inflamatory effect of moxifloxacin (MXF).Methods Obtained human AM from bronchoalveolar lavage and used MTT assay to observe the effects of LTA and MXF on cell activity,optical microscope to investigate the change of the cell morphology,flow cytometry to assess cell apoptosis,RT-PCR to detect the mRNA levels of TLR2,IL-1 β,IL-8 and TNF-α,ELISA for the production of IL-8 to exam RT-PCR.Results LTA showed cytotoxicity on AM in a dose-dependent manner ( P＜0.05 ) ; MXF inhibited the effect of LTA without cytotoxicicy ( P＜0.05 ).LTA promoted apoptosis ( P＜0.05 ) and the mRNA expressions of TRL2,IL-1 β,IL-8 and TNF-α significantly in AM (P＜0.05),the peaks and peak time ofthe above factors were (3.56±0.03) at 12 h,(46.63±7.06) at 6 h,(28.07±1.24) at 12 h and (2.34 ±0.50) at 3 h respectively and increased the release of IL-8 protein level at 24 h (P＜0.05).MXF inhibited the cell apoptosis and the above mRNA expression at 12h ( P＜0.05 ),and inhibited the IL-8 protein level at 24 h( P＜0.05 ).Conclusion LTA showed cytotoxicity on AM,induced AM apoptosis and increased the expression of TLR2,IL-I β,IL-8 and TNF-α of AM ; MXF could protect AM through inhibiting of the above effects and may play a key role beside bactericidal effect in gram-positive bacteria pneumonia.

Imunocitoquímica e imunohistoquímica para o diagnóstico laboratorial da leishmaniose tegumentar / Immunocytochemistry and immunohistochemistry for laboratorial diagnosis of cutaneous leishmaniasis

0 aumento significativo do número de casos notificados da Leishmaniose Tegumentar Americana (LTA) e a expansão geográfica da endemia tem motivado o desenvolvimento de novas tecnologias para auxiliar no diagnóstico das leishmanioses, visando minimizar as restrições apresentadas pelos testes diagnósticos disponíveis nos serviços de saúde. 0 presente trabalho empregou imunocitoquimica e imunohistoquímica (ICQ/IHQ) como métodos diagnósticos laboratoriais para LTA. Amostras de culturas de Leishmania in vitro e cortes histológicos de lesões em animais infectados experimentalmente foram submetidos a ICQ/IHQ, utilizando anticorpos policlonais desenvolvidos para este estudo e o complexo avidina-biotina modificado (Ultra Streptavidin®). Em comparação com outras técnicas empregadas para o diagnóstico da LTA, nos casos avaliados, a IHQ apresentou resultados semelhantes aos da histopatologia com coloração HE, com sensibilidade de 33,3% para formas amastigotas. Quando considerada a presenva de antígenos de Leishmania no padrão celular, a IHQ apresentou uma sensibilidade de 83,3%, significativamente maior que na histopatologia e compatível com metodos padrão ouro de cultura e PCR. As metodologias de ICQ/IHQ desenvolvidas neste trabalho foram capazes de demonstrar em biópsias de lesões, a presença de formas amastigotas e antígenos de Leishmania, oferecendo contribuição adicional ao diagnóstico da LTA, sendo de facil aplicação e podendo ser utilizada no sistema público de saúde.

The significant increase in cutaneous leishmaniasis (CL) notified cases and the geographic expansion of this endemy has motivated the development of new techniques to help in leishmaniasis diagnosis, seeking the minimization of the restrictions imposed by the diagnostic tests available at the health services. The current study applied immunocytochemistry and immunohystochemistry methods (ICC/IHC) for laboratory diagnosis of CL. Imprints and histological sections from tissue infected with Leishmania were submitted to ICC/IHC methods using polyclonal antibodies developed for this study and a modified avidin-biotin complex (Ultra Streptavidin®). The samples also were submitted for routinely stained hematoxylin and eosin (H&E) specimens and gold standard methods (culture and PCR). Compared with other useful techniques for the CL diagnosis, ICC/IHC showed the same sensitivity results (33%) as H&E stain for amastigotes recognition. When the presence of Leishmania antigens was evaluated, ICC/IHC presented 83,3% sensitivity, i.e., higher than that detected by histopathology and equivalent with gold standard methods (culture and PCR). The ICC/IHC techniques developed in the current study were able to recognize amastigote forms and also Leishmania antigens in lesion biopsies, offering an additional help to CL diagnosis and it can be easily applied in the public health system.

Effects of LTA on lung function in patients with stable chronic obstructive pulmonary disease / 中国基层医药

Objective To evaluate the effect of LTA on lung function in stable chronic obstructive pulmonary disease. Methods A randomized and double blind placebo-controlled clinical trail was conducted in 84 patients with COPD. 42 patients were treated by LTA and 42 patients were treated by placebo for 12 weeks. Results LTA provided significant improvement in inspiratory capacity(IC) ,forced expiratory volume in one second(FEV) and forced vital capacity(FVC). Conclusion LTA is an effective bronchodilator on improving lung function and health status in patients with stable COPD.