Effect of cytotoxic T-lymphocyte antigen-4,TNF-alpha polymorphisms on osteosarcoma: evidences from a meta-analysis

Objective： Previous studies have investigated the role of cytotoxic T-lymphocyte antigen-4 （CTLA-4） and tumor necrosis factor-alpha （TNF-a） in carcinogenesis of osteosarcoma, but their results were inconsistent. We aimed to clarify the associations between CTLA-4, TNF-a polymorphism and osteosarcoma risk by using meta-analysis. Methods： We searched relevant studies without language restriction in PubMed, EMbase, Cochrane Library, Google Scholar databases, Chinese National Knowledge Infrastructure （CNKI） and conference literature in humans published prior to March 2013. The strengths of the associations between genetic variants and osteosarcoma risk were estimated by odds ratio （OR） with 95% confidence interval （95% CI）. Results： A total of seven studies with 1,198 osteosarcoma patients and 1,493 controls were selected. Four studies were eligible for CTLA-4 （1,003 osteosarcoma and 1,162 controls）, and three studies for TNF-a （195 osteosarcoma and 331 controls）. Pooled results showed that rs231775 polymorphism of CTLA-4 was associated with osteosarcoma risk （GG vs. AA： OR=1.63, 95% CI=1.24-2.13; GG ＋ GA vs. AA： OR=1.56, 95% CI=1.21-2.01; AA ＋ GA vs. GG： OR=0.83, 95% CI=0.71-0.97; G vs. A： OR=1.21, 95% CI=1.08-1.36）. No significant heterogeneity was observed across the studies. No significant associations were found between rs5742909 polymorphism of CTLA-4 or rs1800629 polymorphism of TNF-a and osteosarcoma risk. Conclusions： These results suggest that the rs231775 polymorphism of CTLA-4 may play an important role in carcinogenesis of osteosarcoma.

No association of the cytotoxic T-lymphocyte associated gene CTLA4 +49A/G polymorphisms with Crohn's disease and ulcerative colitis in Hungarian population samples

瞄准：当前的工作的目标细胞毒素的 T 淋巴细胞抗原是分析 +49A/G 的流行变体的在有 Crohn 的匈牙利病人的 4 基因(CTLA4 )?ˉs 疾病(CD ) 和 ulcerative (UC ) 。方法：有 CD 的 130 个无关的题目的一个总数并且 150 与 UC，和 170 匹配的控制是为单个核苷酸多型性(SNP ) 的 genotyped。遗传型被使用 PCR/RFLP 测试决定。结果：G 等位基因频率和 GG 遗传型的流行在 CD 组，是 38.1% 和 12.3%40.6% 和 18.6% 在 UC 病人，并且 37.4% 和 15.9% 在控制组分别地。结论：当前的学习的结果显示出 +49G SNP 的那辆马车在异质接合或不在匈牙利人口为 CD 或为 UC 在同型结合的形式授与风险任何一个。

Cytotoxic T-lymphocyte antigen 4 gene polymorphisms and susceptibility to chronic hepatitis B

瞄准：在细胞毒素的T淋巴细胞抗原 4 以内估计三个多型性区域( CTLA-4 )基因，在倡导者区域 -318 的 C/T 基础交换( CTLA-4 -318C/T),在 1 个位置上的前的 A/G 替换 49 ( CTLA-4 49A/G )，在 1172 的 T/C 替换(在有长期的肝炎 B 的病人的 CTLA-4 -1172T/C)。方法：当他们介绍了给肝的诊所，有长期的肝炎 B 感染和 150 个健康题目的 51 个病人顺序被招募。长期的肝炎 B (HBV ) 的分类感染了病人作为无征状的带菌状态(26 个病人) 和长期的肝炎 B (25 个病人) 。Genomic DNA 用 Milleros 腌外面方法从凝结反的外部血 Buffy 被孤立。CTLA-4 基因多型性的存在用聚合酶链反应扩大被决定倔强的变化系统(手臂) 。结果：我们观察了在 -318 遗传型频率之间的一个重要协会(T+C- ， T+C+ ， T-C+) 并且危险性到长期的肝炎 B (P=0.012， OR=0.49， 95%CI：0.206-1.162 ) 。然而，我们没为 +49 遗传型频率观察一个重要协会(T+C+ ， T+C-T-C+) 并且 -1172 遗传型频率(C+T+ ， T+C-C+T-) 并且疾病的状态。结论：我们的结果建议 CTLA-4 基因多型性可以部分涉及危险性到长期的肝炎 B。

A Comparative Study on the Effect of BCG-PSN and Thymopeptides on T-lymphocyte Subsets of Normal and Immunosuppressed Mice

To compare the effects of polysaccharide nucleic acid fraction of bacillus calmette guerin (BCG PSN) and thymopeptides on T lymphocytes of normal and immunosuppressed mice, CD4 + and CD8 + T lymphocyte subsets of single nucleic cell in thymus, spleen and peripheral blood were detected successively by flow cytometry after application of BCG PSN and thymopeptides. Meanwhile, CD4 +/CD8 + ratio was also calculated. The results showed that both BCG PSN and thymopeptides could decrease the proportion of CD4 + CD8 + T lymphocyte subsets in the thymus, at the same time increase CD4 + T lymphocyte, CD8 +T lymphocyte proportion in the three tissues. The fluctuation in amplitude was greater in thymopeptides group than that in BCG PSN group. It is concluded that acting location of thymopeptides is in thymus, its stimulating action is stronger than that of BCG PSN, while BCG PSN not only accelerates the differentiation in thymus, but also has some direct stimulation to peripheral CD4 +T lymphocytes, and can maintain CD4 +/CD8 + ratio within normal range. So, BCG PSN is safer.

Antibacterial mechanism with consequent cytotoxicity of different reinforcements in biodegradable magnesium and zinc alloys: A review

Benefits achieved by the biodegradable magnesium(Mg) and zinc(Zn) implants could be suppressed due to the invasion of infectious microbial, common bacteria, and fungi. Postoperative medications and the antibacterial properties of pure Mg and Zn are insufficient against biofilm and antibiotic-resistant bacteria, bringing osteomyelitis, necrosis, and even death. This study evaluates the antibacterial performance of biodegradable Mg and Zn alloys of different reinforcements, including silver(Ag), copper(Cu), lithium(Li), and gallium(Ga). Copper ions(Cu^(2+)) can eradicate biofilms and antibiotic-resistant bacteria by extracting electrons from the cellular structure. Silver ion(Ag^(+)) kills bacteria by creating bonds with the thiol group. Gallium ion(Ga^(3+)) inhibits ferric ion(Fe^(3+)) absorption, leading to nutrient deficiency and bacterial death. Nanoparticles and reactive oxygen species(ROS) can penetrate bacteria cell walls directly, develop bonds with receptors, and damage nucleotides. Antibacterial action depends on the alkali nature of metal ions and their degradation rate, which often causes cytotoxicity in living cells. Therefore, this review emphasizes the insight into degradation rate, antibacterial mechanism, and their consequent cytotoxicity and observes the correlation between antibacterial performance and oxidation number of metal ions.

Echinoside A from Pearsonothuria graeffei Exert the Cytotoxicity to MDA-MB-231 Cells via Mitochondrial Membrane and Modulation of PI3K/Akt/mTOR Pathway

A kind of triterpene glycosides echinoside A(EA)was extracted from sea cucumber Pearsonothuria graeffei,and its yield was about 0.78%.The purity of EA was 99.0%,and its molecular weight was 1206 Da.EA was a linear tetrasaccharide attached to a pentacyclic triterpene aglycon.It inhibited the growth of MDA-MB-231 cells in vitro.The antitumor effect was related to elevate ROS level,decrease mitochondrial membrane potential,enhance caspase-3 expression,induce cells apoptosis and arrest cell cycle at G2/M phase.EA also dose-dependently suppressed the expressions of phophorylation proteins p-PI3K,p-Akt,and p-mTOR as analyzed by western blotting.These results suggested that EA caused MDA-MB-231 cells apoptosis via intrinsic mitochondrial and PI3K/Akt/mTOR pathway.EA can be a potential anti-breast cancer agent to enhance the clinical efficacy.

Association between the cytotoxic T-lymphocyte antigen-4 polymorphisms and breast cancer risk and prognosis

Aim:The aim was to evaluate the potential infl uences of cytotoxic T-lymphocyte antigen-4(CTLA-4)gene polymorphisms on breast cancer risk,the distribution of CTLA-4 single nucleotide polymorphisms(1661AG)in breast cancer patients and control subjects was investigated.Methods:In this case-control study,100 patients with breast cancer as case group and 100 healthy participants as a control group were compared.Genotypes were determined by the polymerase chain reaction-restriction fragment length polymorphism method.Demographic characteristics of the study population,as well as tumor size,tumor grade and stage were collected in a questionnaire designed for this study.The collected data were statistically analyzed by SPSS-16.0(SPSS Inc.,Chicago,USA)predictive analytic software using the Chi-square test.Results:The mean age of women was 43.42±13.1 years.The AA genotype was frequent in case group(43%)whereas the AG genotype was found more in the control group(69%).There was no signifi cant relationship between the studied polymorphisms and the grade,stage and size of the tumor,nor between the studied polymorphisms and estrogen receptor,progesterone receptor and lymph node involvement(P>0.05).Signifi cant association between the studied polymorphisms and breast cancer metastases was found(P=0.02).Conclusion:According to the results of the study,the AA genotype is associated with breast cancer,but none of the studied gene polymorphisms is associated with prognostic factors such as tumor stage,grade or size.

Analysis of Epstein-Barr viral DNA load, EBV-LMP2 specific cytotoxic T-lymphocytes and levels of CD4^＋CD25^＋ T ceils in patients with nasopharyngeal carcinomas positive for IgA antibody to EBV viral capsid antigen

Background Epstein-Barr virus （EBV） is a herpesvirus commonly associated with several malignant diseases including nasopharyngeal carcinoma （NPC）, which is a common cancer in Southeastern Asia. Previous studies showed that plasma levels of EBV-DNA might be a sensitive and reliable biomarker for the diagnosis, staging and evaluating of therapy for NPC. There are a few analyses of the levels of EBV-latent membrane protein 2 （LMP2）-specific cytotoxic T-lymphocytes （CTLs） in patients with NPC. This study was conducted to investigate the levels of EBV-LMP2-specific CTLs, EBV-DNA load and the level of CD4^＋CD25^＋T cells in such patients. Methods From February 2006 to April 2006, 62 patients with NPC, 40 healthy virus carriers positive for EBV viral capsid antigen （EBV-IgA-VCA） and 40 controls were enrolled in the study. We used a highly sensitive ELISPOT assay, real-time polymerase chain reaction （PCR） and flow cytometry to measure the EBV-LMP2-specific CTL response, the EBV DNA load and the level of CD4^＋CD25^＋T cells, respectively. Results The EBV-LMP2-specific CTL responses of the samples from the control, healthy virus carriers and patients with NPC were significantly different from the LMP2 epitopes, with the control and healthy virus carrier samples displaying a stronger response in three cases. There were significant differences in EBV DNA load in serum between NPC and the healthy groups; patients with NPC at stages Ⅲ or Ⅳ had significantly higher viral loads compared with those at stages Ⅰ or Ⅱ. A significantly higher percentage of CD4^＋CD25^＋ T lymphocytes were detected in the patients, compared with healthy virus carriers and healthy controls. Moreover, patients with advanced stages of NPC （Ⅲ and Ⅳ） had significantly higher percentages than the patients with early stages （Ⅰ and Ⅱ）. Conclusions Patients with NPC are frequently unable to establish or maintain sufficient immunosurveillance to control proliferating B cells harboring EBV and to destroy the tumor cells that express immunodominant LMP2 proteins. Controlling the activity of CD4^＋CD25^＋T cells and elevating CD8^＋ cells specific for LMP2 epitopes could be an effective immunotherapy for patients with NPC.

Distribution of natural killer cells and T-lymphocyte subsets in peripheral blood,gallbladder cancer and surrounding tissue

BACKGROUND: The patient with malignant tumor always show immunologic function drawback and ingravescent with tumor development, especially in the aspect of cell-mediated immunity. This study was undertaken to define the relationship between the immune function of local cells and cancer development by investigating the distribution of natural killer (NK) cells and T-lymphocyte subsets in peripheral blood, the cancer tissue and the tissue surrounding gallbladder carcinoma. METHODS: The numbers of CD4(+) and CD8(+) T-lymphocytes and NK cells were measured by flow cytometry in samples taken from gallbladder cancer tissue, the surrounding tissues and peripheral blood of 38 patients, and compared with the numbers in the peripheral blood and gallbladder tissue of 30 patients with cholecystitis as controls. RESULTS: The numbers of CD4(+) and CD8(+) T-cells and NK cells in gallbladder cancer tissues were significantly higher than those in the surrounding tissue and gallbladder with gallstone. However, the ratio of CD4(+)/CD8(+) was lower in the cancer tissue than that in the surrounding tissue and tissue from gallbladders with gallstones. The distribution of CD4(+) and CD8(+) T-cells and NK cells in mucous membrane of cholecystitis gallbladder and that in the tissue surrounding gallbladder cancer were significantly different. CONCLUSIONS: Disproportionate and imbalanced distribution of NK cells and subsets of T-lymphocytes occurs in the mucous membrane proper of gallbladder cancer and surrounding tissue. Although gallbladder cancer tissue has higher expressions of CD4(+), CD8(+) and NK cells, the immune function is low or in an inhibited state. In gallbladder cancer immunization therapy, local cellular immunological function should be enhanced and the protective barrier improved.

Neodymium Oxide Induces Cytotoxicity and Activates NF-κB and Caspase-3 in NR8383 Cells

We investigated whether Nd_2O_3 treatment results in cytotoxicity and other underlying effects in rat NR8383 alveolar macrophages.Cell viability assessed by the MTT assay revealed that Nd_2O_3 was toxic in a dose-dependent manner, but not in a time-dependent manner. An ELISA analysis indicated that exposure to Nd203 caused cell damage and enhanced synthesis and release of inflammatory chemokines. A Western blot analysis showed that protein expression levels of caspase-3, nuclear factor-KB （NF-KB） and its inhibitor IKB increased significantly in response to Nd203 treatment. Both NF-KB and caspase-3 signaling were activated, suggesting that both pathways are involved in Nd203 cytotoxicity.

Microwave-induced Apoptosis and Cytotoxicity of NK Cells through ERK1/2 Signaling

Objective To investigate microwave-induced morphological and functional injury of natural killer（NK） cells and uncover their mechanisms. Methods NK-92 cells were exposed to 10, 30, and 50 m W/cm^2 microwaves for 5 min. Ultrastructural changes, cellular apoptosis and cell cycle regulation were detected at 1 h and 24 h after exposure. Cytotoxic activity was assayed at 1 h after exposure, while perforin and NKG2 D expression were detected at 1 h, 6 h, and 12 h after exposure. To clarify the mechanisms, phosphorylated ERK（p-ERK） was detected at 1 h after exposure. Moreover, microwave-induced cellular apoptosis and cell cycle regulation were analyzed after blockade of ERK signaling by using U0126. Results Microwave-induced morphological and ultrastructural injury, dose-dependent apoptosis（P 〈 0.001） and cell cycle arrest（P 〈 0.001） were detected at 1 h after microwave exposure. Moreover, significant apoptosis was still detected at 24 h after 50 m W/cm^2 microwave exposure（P 〈 0.01）. In the 30 m W/cm^2 microwave exposure model, microwaves impaired the cytotoxic activity of NK-92 cells at 1 h and down regulated perforin protein both at 1 h and 6 h after exposure（P 〈 0.05）. Furthermore, p-ERK was down regulated at 1 h after exposure（P 〈 0.05）, while ERK blockade significantly promoted microwave-induced apoptosis（P 〈 0.05） and downregulation of perforin（P 〈 0.01）. Conclusion Microwave dose-dependently induced morphological and functional injury in NK-92 cells, possibly through ERK-mediated regulation of apoptosis and perforin expression.

Bio-compatibility and cytotoxicity studies of water-soluble CuInS_2-ZnS-AFP fluorescence probe in liver cancer cells

BACKGROUND： The oncogenesis of hepatocellular carcinoma（HCC） is not clear. The current methods of the pertinent studies are not precise and sensitive. The present study was to use liver cancer cell line to explore the bio-compatibility and cytotoxicity of ternary quantum dots（QDs） probe and to evaluate the possible application of QDs in HCC.METHODS： CuInS_2-ZnS-AFP fluorescence probe was designed and synthesized to label the liver cancer cell HepG 2. The cytotoxicity of CuInS_2-ZnS-AFP probe was evaluated by MTT experiments and flow cytometry. RESULTS： The labeling experiments indicated that CuInS_2-ZnS QDs conjugated with AFP antibody could enter HepG 2 cells effectively and emit intensive yellow fluorescence by ultraviolet excitation without changing cellular morphology. Toxicity tests suggested that the cytotoxicity of CuInS_2-ZnS-AFP probe was significantly lower than that of CdT e-ZnS-AFP probe（t test, F=0.8, T=-69.326, P〈0.001）. For CuInS_2-ZnS-AFP probe, timeeffect relationship was presented in intermediate concentration（〉20%） groups（P〈0.05） and dose-effect relationship was presented in almost all of the groups（P〈0.05）. CONCLUSION： CuInS_2-ZnS-AFP QDs probe had better biocompatibility and lower cytotoxicity compared with CdT e-ZnS-AFP probe, and could be used for imaging the living cells in vitro.

Effects of Intraoperative Administration of Dexmedetomidine on the Percentage of T-Lymphocyte Subsets and Natural Killer Cells in Patients with Colorectal Cancer

Study Objective: To observe the effect of dexmedetomidine (DEX) on T-lymphocyte subsets and natural killer (NK) cells in the peripheral blood of perioperative patients with colorectal cancer. Design: A random double-blind control clinical study. Setting: A university hospital. Patients: Forty patients with colorectal cancer, ASA I-П. Interventions: All patients were randomly divided into a DEX group (n = 20) and a control group (n = 20). Before induction of anesthesia, epidural catheters were placed in the L1-L2 or T12-L1 intervertebral spaces. The DEX group received 1 μg/kg of DEX (200 μg/50 ml) intravenously for 15 min prior to the surgery, which was then infused at a rate of 0.5 μg/kg/h until 30 min before the end of the surgery. The control group received an intravenous infusion of saline (50 ml) instead of DEX during the same periods as the DEX group. All patients received routine anesthesia and postoperative analgesia. Measurements: Blood samples from all patients were collected at the following time points: before anesthesia (T0), 24 h after surgery (T1), 48 h after surgery (T2) and 72 h after surgery (T3). Changes in T-lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) and NK cells were determined by flow cytometry. Main Results: Compared with the control group, the percentages of CD3+ and CD4+ cells and the CD4+/CD8+ ratio in the DEX group increased significantly from T1 to T3

DNA Interaction and Cytotoxic Activity of a Chiral Amino-alcohol Schiff Base Derived Cu(Ⅱ) Complex

A novel copper（Ⅱ） complex based on chiral amino-alcohol derived Schiff base ligand,[Cu_4（R-L）_4（H_2O）_2]·（CH_3COOH）_2·（H_2O）（1,（R）-H_2 L =（R）-3-phenyl-2-（2-hydroxy-3-methoxybenzylideneamino） propane-1-ol）,was synthesized and characterized by EA,IR,UV-Vis,ESI-MS,circular dichroism spectra and single-crystal X-ray diffraction.Complex 1 crystallizes in orthorhombic,space group Ρ2_12_12 with a = 15.7660（14）,b = 49.526（3）,c = 10.4213（9） A,V = 8137.2（12） A^3,Ζ = 4,C_（72）H_（81）Cu_4N_4O_（19）,Mr = 1560.57,μ = 1.096 mm^-1,F（000） = 3244,Flack = 0.06（3）,the final R = 0.0924 and w R = 0.2451（I 〉 2σ（I）） for 41108 observed reflections.The interactions of the complex with calf thymus DNA（CT-DNA） were investigated by some spectroscopic technique methods.The results show the complex exhibits strong binding with CT-DNA.In addition,in vitro cytotoxicity test of 1 towards four kinds of human cancerous cell lines（He La,HL-60,Caco-2 and A549） showed substantial cytotoxic activity.The experimental investigations indicated that the chirality of complex 1 play an important role in cytotoxicity and interactions with DNA.

Synthesis,Crystal Structure and Cytotoxic Activities of 1-(Prop-2-yn-1-yl)-7,8-dihydro-1H-benzo[d][1,3]-thiazine-2,5(4H,6H)-dione Derivatives

The important synthetic precursor（Ⅲ）, 1-（prop-2-yn-1-yl）-7,8-dihydro-1Hbenzo[d][1,3]thiazine-2,5（4H,6H）-dione（C（11）H（11）NO2S）, was prepared through a three-component reaction, which was further transferred into cytotoxic triazoles by alkylation and ＂click＂ synthesis in satisfactory yields of 87%^95%. Their structures were characterized by IR, H-RESI-MS and NMR analysis. Meanwhile, the crystal of Ⅲ was obtained and determined by X-ray single-crystal diffraction. Crystal data： orthorhombic system, space group P212121, a = 5.189（4）, b = 8.661（6）, c = 23.498（17） A, V = 1056.2（13） A^3, Z = 4, F（000） = 464, Dc = 1.392 g/cm^3, μ =0.284 mm^-1, R = 0.0637 and wR = 0.1668 for 8182 independent reflections（R（int） = 0.1580） and 2166 observed ones（I 〉 2σ（I））.

Pretreatment of Chrysotile With Rare Earth Compounds Lowered Its Cytotoxicity by Lessening Surface Charges

Pathological effects of asbestos are probably dependent on the special surface properties of the fibers, such as surface charge, surface metal ions. The present study was designed to determine whether the pretreatment of chrysotile asbestos fibers (CAF) with rare earth compounds (REC) solution can reduce their pathogenicity. The results showed that REC-pretreated CAF induced less nitrogen oxide (NO) production by alveolar macrophages (AM). In addition, the pretreatment lowered the capacity of hemolysis and the methylene blue (MB) adsorption of the native CAF. These findings suggested that the pretreatment of CAF with REC solution reduced the in vitro toxicity of CAF by lessening its surface charges. Nevertheless, the pathogenicity and the carcinogenicity of REC-pretreated CAF in vivo remain to be investigated.

INVESTIGATION OF INDUCING EFFECT OF SPECIFIC CYTOTOXICITY OF CTLS BY ANTIGEN PEPTIDES FROM T LYMPHOCYTIC LEUKEMIA CELLS

Objective: To investigate the characteristics of specific antitumor immunity induced by antigen peptides mixture from T lymphocytic leukemia cells. Method: Antigen peptides mixtures were prepared from different leukemia cell lines and then bound with Hsp70 in vitro. Human peripheral blood mononuclear cells (PBMC) were cultured in vitro, and activated with Hsp70-antigen peptides. The activated PBMC was cultured continuously in vitro, and used as effector cells in vitro test of cytotoxicity to different target cells. Results: The antigen peptides from different leukemia cell lines were peptides mixture and could activate PBMC effectively if they were presented by Hsp70. The activated PBMC could proliferate in the presence of IL-2 and Hsp70-antigen peptides. The proliferative PBMC had specific cytotoxicity to leukemia cells corresponding to the antigen peptides. PBMC activated by antigen peptides from T lymphocytic leukemia cell lines could effectively kill T lymphocytic leukemia cells, and the cytotoxicity of these PBMC to T lymphocytic leukemia cells was significantly stronger than that of PBMC activated by antigen peptides from other leukemia cells (P < 0.05). PBMC activated by either Hut78-peptides or Molt 4-peptides could effectively kill Jurkat cells. And the cytotoxicity of PBMC activated by Hut78/Molt-4-peptides to Jurkat cells was significantly stronger than that of PBMC activated by either Hut78-peptides or Molt-4-peptides alone (P<0.05). Conclusion: Antigen peptides mixture from T lymphocytic leukemia cell lines can induce specific cytotoxic effect to T lymphocytic leukemia cells. There exists cross-reactivity among antigen peptides mixture from different T lymphocytic leukemia cell lines. The cross-reactivity could be amplified by blending of different antigen peptides from different T lymphocytic leukemia cell lines, suggesting that it is possible to prepare broad-spectrum antigen peptide vaccine against T lymphocytic leukemia by using multiple leukemia cell lines.

Cytotoxicity of pilocarpine to human corneal stromal cells and its underlying cytotoxic mechanisms

AIM： To examine the cytotoxic effect of pilocarpine, an anti-glaucoma drug, on human corneal stromal（HCS）cells and its underlying cytotoxic mechanisms using an in vitro model of non-transfected HCS cells.· METHODS： After HCS cells were treated with pilocarpine at a concentration from 0.15625 g/L to 20.0 g/L,their morphology and viability were detected by light microscopy and MTT assay. The membrane permeability,DNA fragmentation and ultrastructure were examined by acridine orange（AO）/ethidium bromide（EB） double-staining. DNA electrophoresis and transmission electron microscopy（TEM）, cell cycle, phosphatidylserine（PS）orientation and mitochondrial transmembrane potential（MTP） were assayed by flow cytometry（FCM）. And the activation of caspases was checked by ELISA.· RESULTS： Morphology observations and viability assay showed that pilocarpine at concentrations above0.625 g/L induced dose- and time-dependent morphological abnormality and viability decline of HCS cells. AO/EB double-staining, DNA electrophoresis and TEM noted that pilocarpine at concentrations above 0.625 g/L induced dose- and/or time-dependent membrane permeability elevation, DNA fragmentation, and apoptotic body formation of the cells. Moreover, FCM and ELISA assays revealed that 2.5 g/L pilocarpine also induced S phase arrest, PS externalization, MTP disruption, and caspase-8,-9 and-3 activation of the cells.· CONCLUSION： Pilocarpine at concentrations above0.625 g/L（1/32 of its clinical therapeutic dosage） has a dose- and time-dependent cytotoxicity to HCS cells by inducing apoptosis in these cells, which is most probably regulated by a death receptor-mediated mitochondrion-dependent signaling pathway.

Analysis of T-lymphocyte subtypes of the peripheral blood and skindelayed-type hypersensitivity in patients with hyper-IgE syndrome

Objective: To study the function of ce ll ular immunity in patients with hyper-IgE syndrome (HIE). Methods: T-lymphocyte subtypes of the peripheral blood and cutaneous delayed-typ e hypersensitivity (DTH) response to two recall antigens, tetanus toxoid (TT) an d purified protein derivative(PPD), were measured in five patients with HIE and 15 healthy controls, respectively. Results: The CD4+ cell cou nts in HIE group were significantly lower than those in control group (P<0. 01). In contrast, CD8+ cells were significantly higher than those in the contro l (P<0.01). The induration sizes of DTH response to two recall antigens wer e smaller in HIE group than those in the control group (P<0.01). Co nclusion: There was an immunologic dysfunction of T lymphocytes in the p atients with HIE.

Investigation of Antibacterial, Cytotoxic and Antioxidant Properties of the Mangrove Plant Xylocarpus mekongensis

The plant (Xylocarpus mekongensis) of the Sundarbans mangrove origin was evaluated for its antibacterial, cytotoxic and antioxidant properties using methanolic and chloroformic leaf, stem and bark extracts, respectively. The methanolic extracts contained higher amount of total phenolics, flavonoids, tannins than the chloroformic extracts and the result was in correlation with their ferric reducing power ability as well. However, the chloroformic bark extract contained more potent DPPH free radical scavenging activity than others. Antibacterial activity of the extracts was determined against both Gram-positive (Micrococcus and Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Vibrio cholerae, Salmonella typhimurium and Salmonella paratyphi) by disc diffusion assay and their zone of inhibitions (ZOI) were measured. Moreover, their minimum inhibitory concentrations (MIC) were determined by tube dilution method. Chloroformic bark and stem extracts showed strong inhibition to growth of P. aeruginosa (ZOI = 19 mm and MIC = 150 μg/ml) and S. aureus (ZOI = 19.5 mm and MIC = 250 μg/ml), respectively. All six extracts were subjected to brine shrimp lethality bioassay for possible measure of cytotoxicity. Concentration dependent increment in percentage mortality of brine Shrimp nauplii produced by the extracts indicated the presence of cytotoxic principles in these extractives. Therefore, Xylocarpus mekongensis showed antioxidant, antibacterial and cytotoxic activities.