Effect of IFNα-2a on Fas expression and apoptosis rate of peripheral blood cytotoxic T cells in patients with hepatitis B

Interferon(IFN) with antiviral and im-munomodulatory activities is one of the most important therapeutic agents for the treatment of chronic hepatitis. The apoptotic effect of IFN is influenced by cell type and the types of IFN, which suppresses proliferation and induces apoptosis in some cell types while inhibiting apoptosis in others. The aim of this study was to explore the effect of IFNα-2a on Fas expression and the apoptosis rate of peripheral blood cytotoxic T cells (CTLs) in patients with hepatitis B. METHODS:Peripheral blood mononuclear cells were isolated from 26 patients with hepatitis B including 16 patients with chronic hepatitis B and 10 patients with chronic severe hepatitis B. Fas expression and apoptosis rate of CTLs were analyzed with flow cytometry before and after IFNα-2a treatment. RESULTS:Before IFNα-2a treatment, Fas expression and apoptosis rate of CTLs from patients with chronic hepatitis B were significantly higher than those from patients with chronic severe hepatitis B and healthy controls respectively. No significant difference was observed between Fas expression and apoptosis rate of CTLs from patients with chronic severe hepatitis B and healthy controls. After IFNα-2a treatment,Fas expression and apoptosis rate of CTLs from different groups were compared with those before IFNα-2a treatment, showing no significant difference despite alternation of different degree. CONCLUSIONS:Activation induced cell death (AICD) exists in peripheral blood CTLs from patients with hepatitis B. No effect of IFNα-2a exerts on Fas expression and apoptosis rate of Fas in patients with hepatitis B.

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.

Cytotoxic effect and possible mechanisms of Tetracaine on human corneal epithelial cells in vitro

AIM： To demonstrate the cytotoxic effect and possible mechanisms of Tetracaine on human corneal epithelial（HCEP） cells in vitro.·METHODS： In vitro cultured HCEP cell were treated with Tetracaine hydrochloride at different doses for different times, and their morphology, viability, and plasma membrane permeability were detected by light microscopy, methyl thiazolyl tetrazolium（MTT） assay,and acridine orange（AO）/ethidium bromide（EB） staining,respectively. Their cell cycle progression, phosphati-dylserine orientation in plasma membrane, and mitochondrial membrane potential（MTP） were assessed by flow cytometry. DNA fragmentation, ultrastructure,caspase activation, and the cytoplasmic apoptosis inducing factor（AIF） and cytochrome c（Cyt. c） along with the expression of B-cell lymphoma-2（Bcl-2） family proteins were examined by gel electrophoresis,transmission electron microscope, enzyme linked immunosorbent assay（ELISA）, and Western blot,respectively.·RESULTS： After exposed to Tetracaine at doses from10.0 to 0.3125 g/L, the HCEP cells showed dose- and time-dependent morphological abnormality and typical cytopathic effect, viability decline, and plasma membrane permeability elevation. Tetracaine induced phosphati-dylserine externalization, DNA fragmentation, G1 phase arrest, and ultrastructural abnormality and apoptotic body formation. Furthermore, Tetracaine at a dose of0.3125 g/L also induced caspase-3,-9 and-8 activation,MTP disruption, up-regulation of the cytoplasmic amount of Cyt. c and AIF, the expressions of Bax and Bad, and down-regulation of the expressions of Bcl-2 and Bcl-x L.·CONCLUSION： Tetracaine above 0.3125 g/L（1/32 of its clinical applied dosage） has a dose- and time-dependent cytotoxicity to HCEP cells in vitro, with inducing cellapoptosis via a death receptor-mediated mitochondriondependent pathway.

Cytotoxicity of Captafol in Mammalian Cells

The cytotoxicity of captafol, a phthalimide-derived fungicide, was evaluated in IB-RS-2 cells. Captafol at 0. 12-1 .0μg/ml blocks the cell multiplication. This effect is concentrationdependent, only partially rcversible and the degree of inhibition increases with time. The synthesis of DNA and RNA is inhibited in parallel by increasing concentrations of the chemical

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.

Cytotoxic Responses and Apoptosis in Rat Kidney Epithelial Cells Exposed to Lead

The toxic effects of lead on normal rat kidney epithelial cells（NRK cells）may occur via various pathways.However,the role of intrinsic mitochondrial pathway in Lead-induced apoptosis in NRK cells has not been investigated.The purpose of our study was to investigate cytotoxic responses and cell apoptosis mediated by lead in NRK cells.

Recombinant E.coli LLO/OVA Vaccination Effectively Inhibits Murine Melanoma Metastasis to Lung by CD8^+T Cells Immunity

Objective： To construct recombinant E.coli LLO/OVA and investigate its tumor metastatic inhibition effect in B16 OVA melanoma challenged mice. Methods： Recombinant E.coli LLO/OVA was constructed and the expression of listeriolysin O （LLO） and ovalbumin （OVA） of the vaccine was determined by coomassie brilliant blue staining and western blotting, After 3 subcutaneous injections of E.coli LLO/OVA, the percentages of CD3^＋CD4^＋T, CD4^＋CD25^＋T, CD3^CD8^＋T and OVA257-264 SIINFEKL specific CD8^＋T cells were determined by flow cytomytry, and the tumor metastatic inhibition effect in B16 OVA melanoma challenged mice was observed. Results： Recombinant E.coli LLO/OVA was successfully constructed, and the expression of LLO and OVA of the vaccine was confirmed. After 3 subcutaneous injections of E.coli LLO/OVA and E.coli OVA in mice, the percentages of CD3^＋CD4^＋T, CD4^＋CD25^＋T and CD3^＋CD8^＋T cells were equivalent in the two groups of mice. However, there were significantly more OVA257-264 SIINFEKL specific CD8^＋T cells in E.coli LLO/OVA vaccinated mice than that in E.coli OVA vaccinated mice. The prophylactic E.coli LLO/OVA vaccination effectively prevented the tumor metastasis to lungs in B16 OVA melanoma challenged mice. Depletion of CD8^＋T cells significantly impaired the tumor inhibition effect of the vaccine in B16 OVA challenged mice. The therapeutic vaccination of E.coli LLO/OVA significantly prevented melanoma metastasis to lungs in B I6 OVA challenged mice too. Conclusion： Recombinant E.coli LLO/OVA vaccination is highly effective in inhibiting murine malignant melanoma metastasis by promoting CD8^＋T cell immunity.

In vitro acute cytotoxicity of neonicotinoid insecticide imidacloprid to gill cell line of flounder Paralichthy olivaceus

In vitro acute cytotoxicity of neonicotinoid insecticide imidacloprid （IMI） to the gill cell line of flounder （FG） that collected in the gill ofParalichthys olivaceus, was examined by 3 widely used endpoint bioassays： NR （neutral red）, MTT （3-（4,5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazoliurn bromide） and TCP （total cell protein）. The result shows that the IMI increased at concentrations ≥0.5 μg/ml. The ICs0 value of NR, MTT, and TCP was 41.86, 38.46, and 39.08 μg/ml, respectively. The ultrastructural observation revealed that the mitochondria of the cells exposed to 60 μg/ml IMI for 48 h were severely damaged, swollen or disrupted, while their nuclei and rough endoplasmic reticulurn （RER） remained normal. This would suggest that the mitochondria are probably the primary target of IMI.

Cytotoxicity of Modified Nonequilibrium Plasma with Chlorhexidine Digluconate on Primary Cultured Human Gingival Fibroblasts

The aim of this study was to investigate the cytotoxicity of modified nonequilibrium plasma with chlorhexidine digluconate（CHX） on human gingival fibroblasts（HGFs）, and to evaluate the biosecurity of modified nonequilibrium plasma with 2% CHX as a new method of root canal treatment. Tissue samples taken from human gingiva were primarily cultured and passaged. Cells from passages 3–7 were used. HGFs were treated by modified nonequilibrium plasma with 2% CHX for 0 min（control group）, 30 s, 1 min, 1.5 min, 3 min, 5 min, and 10 min, respectively, and then they were incubated for 0, 24, and 48 h. After that, cell counting kit-8（CCK-8） assay was applied to analyze the cytotoxicity of modified nonequilibrium plasma with 2% CHX on HGFs. There was no significant difference between the 0 h group treated with the modified nonequilibrium plasma for 1 min and the control group（P〉0.05）. However, there were significant differences between all the other treated groups and the control group（P〈0.05）. When treated for 1.5 min or shorter, the cell viability was obviously increased; while treated for 3 min or longer, it was obviously reduced. Moreover, when successively cultured for 0, 24, and 48 h, cell viability was decreased at first and then increased in the 3-min-treated and 5-min-treated groups. The modified nonequilibrium plasma with 2% CHX was of no influence on cell viability in 1.5 min treatment, and it could be safely used on root canal treatment.

THE EFFECTS OF HSP27 ON THE CYTOTOXICITY OF RAT EMBRYO FIBROBLAST INDUCED BY CISPLATIN

Objective: To investigate the protective effects of heat shock protein 2700(Hsp27) on cisplatin inducing cytotoxicity in a temperature mutant SV40 large T antigen transformed rat embryo fibroblast (P1-Hsp27). Methods The cytotoxical effects of cisplatin on the proliferation status of P1-Hsp27 cells in the presence or absence of Hsp27 were measured by MTT assay. Results Cisplatin possessed dose-dependent cytotoxicity on P1-Hsp27 cells. 48h after treatment, about 50% cells were dead in those cells exposed to 200μmol cislatin. However, no obvious protective effects of Hsp27 on cisplatin induced cytotoxicity could he observed (P>0. 05), except in those cells exposed to 500μmol cisplatin 12h after treatment.Conclusion Hsp27 has no obvious protective effects on cisplatin inducing cytotoxicity.

Multi‑omics integration identifies regulatory factors underlying bovine subclinical mastitis

Background Mastitis caused by multiple factors remains one of the most common and costly disease of the dairy industry.Multi-omics approaches enable the comprehensive investigation of the complex interactions between mul-tiple layers of information to provide a more holistic view of disease pathogenesis.Therefore,this study investigated the genomic and epigenomic signatures and the possible regulatory mechanisms underlying subclinical mastitis by integrating RNA sequencing data(mRNA and lncRNA),small RNA sequencing data(miRNA)and DNA methylation sequencing data of milk somatic cells from 10 healthy cows and 20 cows with naturally occurring subclinical mastitis caused by Staphylococcus aureus or Staphylococcus chromogenes.Results Functional investigation of the data sets through gene set analysis uncovered 3458 biological process GO terms and 170 KEGG pathways with altered activities during subclinical mastitis,provided further insights into subclin-ical mastitis and revealed the involvement of multi-omics signatures in the altered immune responses and impaired mammary gland productivity during subclinical mastitis.The abundant genomic and epigenomic signatures with sig-nificant alterations related to subclinical mastitis were observed,including 30,846,2552,1276 and 57 differential methylation haplotype blocks(dMHBs),differentially expressed genes(DEGs),lncRNAs(DELs)and miRNAs(DEMs),respectively.Next,5 factors presenting the principal variation of differential multi-omics signatures were identified.The important roles of Factor 1(DEG,DEM and DEL)and Factor 2(dMHB and DEM),in the regulation of immune defense and impaired mammary gland functions during subclinical mastitis were revealed.Each of the omics within Factors 1 and 2 explained about 20%of the source of variation in subclinical mastitis.Also,networks of impor-tant functional gene sets with the involvement of multi-omics signatures were demonstrated,which contributed to a comprehensive view of the possible regulatory mechanisms underlying subclinical mastitis.Furthermore,multi-omics integration enabled the association of the epigenomic regulatory factors(dMHBs,DELs and DEMs)of altered genes in important pathways,such as‘Staphylococcus aureus infection pathway’and‘natural killer cell mediated cyto-toxicity pathway’,etc.,which provides further insights into mastitis regulatory mechanisms.Moreover,few multi-omics signatures(14 dMHBs,25 DEGs,18 DELs and 5 DEMs)were identified as candidate discriminant signatures with capac-ity of distinguishing subclinical mastitis cows from healthy cows.Conclusion The integration of genomic and epigenomic data by multi-omics approaches in this study provided a better understanding of the molecular mechanisms underlying subclinical mastitis and identified multi-omics candidate discriminant signatures for subclinical mastitis,which may ultimately lead to the development of more effective mastitis control and management strategies.

Prognosis and immunological role of HLA-DMA in lung adenocarcinoma

Background:HLA-DMA presents pathogen-derived antigens to CD4+and CD8+T cells,respectively,and plays a significant part in initiating the immune response.So far,the impact of HLA expression on the prognosis of BC cells is controversial,because few studies have shown that the expressions of some HLA genes are related to the improvement of the survival rate.Up till now,however,the relationship between HLA-DMA and LUAD has not yet been assessed.Methods:We analyzed the TCGA database and assessed the prognostic value of HLA-DMA in LUAD.We conducted the Kruskal–Wallis and Wilcoxon signed-rank test and utilized logistic regression to assess the role of clinical-pathologic characteristics and HLA-DMA expression.Kaplan–Meier method and the multivariate and univariate Cox regression were also used for evaluating the prognosis-related factors of LUAD.GSEA was used to identify HLA-DMA-related key pathways.The ssGSEA of the TCGA data was used to investigate the correlations between HLA-DMA and cancer immune cell infiltration.Results:Low HLA-DMA expression was related to poorer disease-specific survival(DSS)and overall survival(OS)of LUAD patients.GSEA revealed that HLA-DMA was tightly interrelated with an immune response by the reactome activation of anterior hox genes in hindbrain development during the early embryogenesis signaling pathway.The expression of HLA-DMA was positively associated with cytotoxic cell infiltration and negatively related to the Th2 cell infiltration according to the ssGSEA.Western blotting and the CCK-8 assay showed that KD-HLA-DMA could significantly increase the proliferation of A549 cells and significantly reduce cell pyroptosis.Conclusion:All the observations implied that HLA-DMA was associated with patient prognosis and immune infiltration in LUAD.

Genetic susceptibility to autoimmune liver disease

Autoimmune hepatitis(AIH),primary sclerosing cholangitis(PSC) and primary biliary cirrhosis(PBC) are considered as putative autoimmune diseases of the liver.Whereas strong evidence that bacterial infection may trigger PBC exists,the etiologies for PSC and AIH remain unknown.Although there have been significant discoveries of genetic polymorphisms that may underlie the susceptibility to these liver diseases,their associations with environmental triggers and the subsequent implications have been difficult to elucidate.While single nucleotide polymorphisms within the negative costimulatory molecule cytotoxic T lymphocyte antigen 4(CTLA-4) have been suggested as genetic susceptibility factors for all three disorders,we discuss the implications of CTLA-4 susceptibility alleles mainly in the context of PBC,where Novosphingobium aromaticivorans,an ubiquitous alphaproteobacterium,has recently been specifically associated with the pathogenesis of this devastating liver disease.Ultimately,the discovery of infectious triggers of PBC may expand the concept of genetic susceptibility in immune-mediated liver diseases from the concept of aberrant immune responses against self-antigens to insufficient and/or inappropriate immunological defense mechanisms allowing microbes to cross natural barriers,establish infection and damage respective target organs.

Adaptive immune response during hepatitis C virus infection

Hepatitis C virus(HCV)infection affects about 170 million people worldwide and it is a major cause of liver cirrhosis and hepatocellular carcinoma.HCV is a hepatotropic non-cytopathic virus able to persist in a great percentage of infected hosts due to its ability to escape from the immune control.Liver damage and disease progression during HCV infection are driven by both viral and host factors.Specifically,adaptive immune response carries out an essential task in controllingnon-cytopathic viruses because of its ability to recognize infected cells and to destroy them by cytopathic mechanisms and to eliminate the virus by non-cytolytic machinery.HCV is able to impair this response by several means such as developing escape mutations in neutralizing antibodies and in T cell receptor viral epitope recognition sites and inducing HCV-specific cytotoxic T cell anergy and deletion.To impair HCV-specific T cell reactivity,HCV affects effector T cell regulation by modulating T helper and Treg response and by impairing the balance between positive and negative co-stimulatory molecules and between pro-and antiapoptotic proteins.In this review,the role of adaptive immune response in controlling HCV infection and the HCV mechanisms to evade this response are reviewed.

In vitro evaluation of inorganic and methyl mercury mediated cytotoxic effect on neural cells derived from different animal species

To extend the current understanding of the mercury-mediated cytotoxic effect,five neural cell lines established from different animal species were comparatively analyzed using three different endpoint bioassays：thiazolyl blue tetrazolium bromide,3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide assay（MTT）,neutral red uptake assay（NRU）,and Coomassie blue assay（CB）.Following a 24-hr exposure to selected concentrations of mercury chloride（HgCl_2） and methylmercury（Ⅱ） chloride（MeHgCl）,the cytotoxic effect on test cells was characterized by comparing their 50%inhibition concentration（IC_（50）） values.Experimental results indicated that both these forms of mercury were toxic to all the neural cells,but at very different degrees.The IC_（50）values of MeHgCl among these cell lines ranged from 1.15±0.22 to 10.31 ± 0.70 μmol/L while the IC_（50） values for HgCl_2 were much higher,ranging from 6.44 ± 0.36 to 160.97±19.63 μmol/L,indicating the more toxic nature of MeHgCl.The IC_（50） ratio between HgCl_2and MeHgCl ranged from 1.75 to 96.0,which confirms that organic mercury is much more toxic to these neural cells than inorganic mercury.Among these cell lines,HGST-BR and TriG44 derived from marine sea turtles showed a significantly high tolerance to HgCl_2 as compared to the three mammalian neural cells.Among these neural cells,SK-N-SH represented the most sensitive cells to both chemical forms of mercury.

The Characterization of Steam Distillation as an Extraction Method to Extract Volatile Compounds from <i>Prunella vulgaris</i>and the Investigation of Their Anti-Tumorous Effect

Prunella vulgaris (PV) is a perennial plant which is widely grown around the world. It has been widely used as a medicinal treatment for generations. Previous studies showed extracts from this plant had a wide range of therapeutic efficacy, including anti-tumorous effect. However, the volatile organic compounds (VOCs) extracted from it were rarely explored. This paper reports on the characterization of a steam distillation process to extract VOCs in PV and also the anti-tumorous effects of the PV distillate using the tetrazolium-based Cell Counting Kit-8 (CCK-8) as the test agent, when the VOCs were used to treat oral squamous cancer cells, SSC154. It was found that most abundant VOCs came out steadily and continuously for as long as the duration of the steam extraction could extend. However, some compounds such as benzaldehyde did show depletion as the distillation process progressed, while some compounds such as caryophyllene oxide was only sparsely found at the beginning of distillation. The PV distillate was mildly effective in its cytotoxicity to cancer cells SCC154, in a dosage dependent manner.

A STUDY OF THE CYTOTOXICITY OF MALIGNANT PLEURAL EFFUSION LYMPHOCYTES AND LAK CELLS AGAINST AUTOLOGOUS TUMOR CELLS

The cytotoxicity of malignant pleural effusion lymphocytes (MPEL) against autologous tumor cells (ATC) were compared with that of peripheral blood lymphocytes (PBL). It was demonstracted that the cytotoxicity of PBL was higher than that of MPEL (P< 0.001), but the cytotoxicity and expansion of MPEL-activated by rIL-2 was much higher than that of PBL activated by rIL-2 (LAK cells) (P< 0.001). This shows that local immune reaction of the pleural cavity of patients with malignant pleural effusion was in the state of suppression. MPEL activated are better effector cells than LAK cells in tumor adoptive immunotherapy.

Necrostatin-1 protection of dopaminergic neurons

Necroptosis is characterized by programmed necrotic cell death and autophagic activation and might be involved in the death process of dopaminergic neurons in Parkinson＇s disease. We hypothesized that necrostatin-1 could block necroptosis and give protection to dopaminergic neurons. There is likely to be crosstalk between necroptosis and other cell death pathways, such as apoptosis and autophagy. PC12 cells were pretreated with necroststin-1 1 hour before exposure to 6-hydroxydopamine. We examined cell viability, mitochondrial membrane potential and expression patterns of apoptotic and necroptotic death signaling proteins. The results showed that the autophagy/lysosomal pathway is involved in the 6-hydroxydopamine-induced death process of PC12 cells. Mitochondrial disability induced overactive autophagy, increased cathepsin B expression, and diminished Bcl-2 expression. Necrostatin-1 within a certain concentration range（5–30 μM） elevated the viability of PC12 cells, stabilized mitochondrial membrane potential, inhibited excessive autophagy, reduced the expression of LC3-II and cathepsin B, and increased Bcl-2 expression. These findings suggest that necrostatin-1 exerted a protective effect against injury on dopaminergic neurons. Necrostatin-1 interacts with the apoptosis signaling pathway during this process. This pathway could be a new neuroprotective and therapeutic target in Parkinson＇s disease.

Monoclonal antibodies that target the immunogenic proteins expressed in colorectal cancer

In an attempt to improve upon the end results obtained in treating colorectal cancer it was apparent that the earlier the diagnosis that could be obtained, the better the chance for obtaining desired results. In the case of more advanced tumors typified by later stage colorectal cancer, surgical debulking is an important part of the treatment strategy. Here the use of additional therapeu-tic modalities including chemotherapy and present day immunotherapy has failed to accomplish the desired im-provements that have been sought after. Adjuvant ther-apy, has offered little to the overall survival. The concept of early detection is now recognized as the initial step in reaching proper end results and can readily be demon-strated from colorectal cancer studies. Here survival has been found to be a reflection of the stage at which the tumor is first identified and treated. When specific mono-clonals targeting colorectal cancer are employed diagnos-tically, we have been able to demonstrate detection of colorectal cancer at its inception as a premalignant lesion, such that genotypic features can be identified before the phenotypic appearance of cancer can be noted.