A rhoptry protein, localizing in the bulb region of rhoptries, could induce protective immunity against Eimeria tenella infection.

Background: Rhoptry organelle proteins (ROPs) secreted by apicomplexan parasites play important roles during parasites invasion and survival in host cells, and are potential vaccine candidates against apicomplexan diseases. Eimeria tenella (E. tenella) is one of the most noteworthy apicomplexan species, which causes hemorrhagic pathologies. Although dozens of putative E. tenella ROP sequences are annotated, most ROP proteins are not well studied. Methods: In this study, an E. tenella ROP21 gene was identified and the recombinant EtROP21 protein (rEtROP21) was expressed in Escherichia coli. The developmental expression levels, localization, and protective efficacy against E. tenella infection in chickens were studied. Results: An EtROP21 gene fragment with an open reading frame (ORF) of 981 bp was obtained from the Beijing strain of E. tenella. The rEtROP21 has a molecular weight of approximately 50 kDa and was recognized by rEtROP21-immunized mouse serum. Two specific protein bands, about 43 KDa and 95 KDa in size, were detected in the whole sporozoite proteins using the rEtROP21-immunized chicken serum. RT-qPCR analysis of the E. tenella ROP21 gene (EtROP21) revealed that its mRNA levels were higher in merozoites and sporozoites than in sporulated and unsporulated oocysts. Immunofluorescence and immunoelectron analyses showed that the EtROP21 protein predominantly localizes in the bulb region of rhoptries distributed at anterior, posterior, and perinuclear regions of E. tenella sporozoites. Immunization and challenge experiments revealed that immunizing chickens with rEtROP21 significantly increased their average body weight gain while decreasing mean lesion score and oocyst output (P <0.05). When compared with the challenged control group, the rEtROP21-immunized group was associated with a significantly higher relative weight gain (90.2%) and a greater reduction in oocyst output (67%) (P <0.05). The anticoccidial index of the rEtROP21-immunized group was 163.2. Chicken serum ELISA revealed that the levels of the specific anti- rEtROP21 antibody, IFN-γ, and IL-4 were significantly higher in the rEtROP21-immunized group than in the challenged control group (P <0.05). Conclusion: These results indicate that rEtROP21 can induce a high level of specific immune response and it is a potential candidate for the development of vaccines against E. tenella infection in chickens.

Molecular characterization and protective efficacy of vacuolar protein sorting 29 from Eimeria tenella.

Introduction: Vacuolar protein sorting 29 (VPS29) is a core component of the retromer-retriever complex and is essential for recycling numerous cell-surface cargoes from endosomes. However, there are no reports yet on VPS29 of Eimeria spp. Methods: Here, we cloned and prokaryotically expressed a partial sequence of Eimeria tenella VPS29 (EtVPS29) with RT-PCR and engineered strain of Escherichia coli respectively. The localization of the VPS29 protein in E. tenella sporozoites was investigated with immunofluorescence (IFA) and overexpression assays. And its protective efficacy against E. tenella infection was investigated in chickens with the animal protection test. Results: An EtVPS29 gene fragment with an ORF reading frame of 549 bp was cloned. The band size of the expressed recombinant protein, rEtVPS29, was approximately 39 kDa and was recognized by the chicken anti-E. tenella positive serum. EtVPS29 protein was observed widely distributing in the cytoplasm of E. tenella sporozoites in the IFA and overexpression assays. rEtVPS29 significantly increased average body weight gain and decreased mean lesion score and oocyst output in chickens. The relative weight gain rate in the rEtVPS29-immunized group was 62.9%, which was significantly higher than that in the unimmunized and challenged group (P < 0.05). The percentage of reduced oocyst output in the rEtVPS29 immunized group was 32.2%. The anticoccidial index of the rEtVPS29-immunized group was 144.2. Serum ELISA also showed that rEtVPS29 immunization induced high levels of specific antibodies in chickens. Discussion: These results suggest that rEtVPS29 can induce a specific immune response and is a potential candidate for the development of novel vaccines against E. tenella infections in chickens.

Investigation on the potential adverse outcome pathway of the sensitive endpoint for nephrotoxicity induced by gardenia yellow based on an integrated strategy using bioinformatics analysis and in vitro testing validation.

To explore the potential the adverse outcome pathway of Gardenia Yellow (GY)-induced sensitive endpoint for nephrotoxicity, an integrated strategy was applied in the present study. Using bioinformatic analysis, based on the constructed Protein-protein interaction networks, Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis on the core target network were performed to illustrate the potential gene targets and signal pathways. Then, the most enriched pathway was validated with Cell counting kit-8 assays and Western blot analysis in embryonic kidney epithelial 293 cell models. According to the findings, GY may interact with 321 targets related to the endpoint. The five targets on the top ranking in the PPI network were STAT3, SRC, HRAS, AKT1, EP300. Among them, PI3K/Akt was the most enriched pathway. In vitro testing showed that GY exerted a proliferative effect on the cell variability in a dose-dependent manner. GY at concentration of 1000 µg/ml and stimulation for 30 min can significantly enhance the expression of phosphorylated Akt. Thus, after the quantitative weight of evidence evaluation, Akt phosphorylation induced PI3K/Akt activation was speculated as a molecular initiating event leading to a proliferative and inflammatory response in renal tubular epithelial cells.

Enhancing Immune Responses to a DNA Vaccine Encoding Toxoplasma gondii GRA7 Using Calcium Phosphate Nanoparticles as an Adjuvant.

Toxoplasma gondii infects almost all warm-blooded animals, including humans. DNA vaccines are an effective strategy against T. gondii infection, but these vaccines have often been poorly immunogenic due to the poor distribution of plasmids or degradation by lysosomes. It is necessary to evaluate the antigen delivery system for optimal vaccination strategy. Nanoparticles (NPs) have been shown to modulate and enhance the cellular humoral immune response. Here, we studied the immunological properties of calcium phosphate nanoparticles (CaPNs) as nanoadjuvants to enhance the protective effect of T. gondii dense granule protein (GRA7). BALB/c mice were injected three times and then challenged with T. gondii RH strain tachyzoites. Mice vaccinated with GRA7-pEGFP-C2+nano-adjuvant (CaPNs) showed a strong cellular immune response, as monitored by elevated levels of anti-T. gondii-specific immunoglobulin G (IgG), a higher IgG2a-to-IgG1 ratio, elevated interleukin (IL)-12 and interferon (IFN)-γ production, and low IL-4 levels. We found that a significantly higher level of splenocyte proliferation was induced by GRA7-pEGFP-C2+nano-adjuvant (CaPNs) immunization, and a significantly prolonged survival time and decreased parasite burden were observed in vaccine-immunized mice. These data indicated that CaPN-based immunization with T. gondii GRA7 is a promising approach to improve vaccination.

The 2019 novel coronavirus resource.

An ongoing outbreak of a novel coronavirus infection in Wuhan, China since December 2019 has led to 31,516 infected persons and 638 deaths across 25 countries (till 16:00 on February 7, 2020). The virus causing this pneumonia was then named as the 2019 novel coronavirus (2019-nCoV) by the World Health Organization. To promote the data sharing and make all relevant information of 2019-nCoV publicly available, we construct the 2019 Novel Coronavirus Resource (2019nCoVR, https://bigd.big.ac.cn/ncov). 2019nCoVR features comprehensive integration of genomic and proteomic sequences as well as their metadata information from the Global Initiative on Sharing All Influenza Data, National Center for Biotechnology Information, China National GeneBank, National Microbiology Data Center and China National Center for Bioinformation (CNCB)/National Genomics Data Center (NGDC). It also incorporates a wide range of relevant information including scientific literatures, news, and popular articles for science dissemination, and provides visualization functionalities for genome variation analysis results based on all collected 2019-nCoV strains. Moreover, by linking seamlessly with related databases in CNCB/NGDC, 2019nCoVR offers virus data submission and sharing services for raw sequence reads and assembled sequences. In this report, we provide comprehensive descriptions on data deposition, management, release and utility in 2019nCoVR, laying important foundations in aid of studies on virus classification and origin, genome variation and evolution, fast detection, drug development and pneumonia precision prevention and therapy.

[Experimental study of TGF-ß2 antisense oligonucleotide on inhibiting corneal scar hyperplasia in rabbits].

OBJECTIVE: To investigate the influence of TGF-ß2 antisense oligonucleotide (ASON) on preventing corneal scar hyperplasia in rabbits and to provide experimental evidence for its clinical application. METHODS: It was an experimental study. One hundred and ninety two New Zealand white rabbits were randomly divided into 4 groups (groups A, B, C and D). Corneal injury models were established in all groups. There were 48 experimental animals in each group. TGF-ß2 ASON was dropped into right eyes in group A, dexamethasone was dropped into right eyes in group B, deionized water was dropped into right eyes in group C and nothing was dropped into right eyes in group D after the operation. The corneas were surgically removed and assessed by hematoxylin eosin (HE) staining, immunohistochemical study and real time PCR at four different time points (4 d, 7 d, 14 d and 28 d) after surgery. RESULTS: HE staining: at the same time point, fibroblasts in the groups A and B were significantly fewer than that in the groups C and D, the difference was statistically significant (P < 0.05), but there was no significant difference between groups A and B or groups C and D. Immunohistochemical observation found that the expression of α-smooth muscle actin (α-SMA) positive fibroblasts could be observed by the 4th day (9.44 ± 0.47/HP), reached a climax by the 7th day (12.50 ± 0.81/HP), and returned to the baseline levels by the 14th day (0.85 ± 0.43/HP) in the group A, which was similar to that in the group B (9.49 ± 0.95, 12.42 ± 0.70, 0.86 ± 0.79/HP) at the same time point (P > 0.05), but it was significantly fewer than that in the group C(20.14 ± 0.78, 18.19 ± 1.28, 4.87 ± 0.58/HP) and group D(20.21 ± 0.92, 18.25 ± 1.39, 5.00 ± 2.217/HP), which was statistical significant (P < 0.05). The staining intensity of fibronectin (FN) in groups A and B was significantly weaker than that in groups C and D. Real time PCR analysis showed that at each time point, the expression of TGF-ß2 mRNAs in groups A and B was significantly lower than that in groups C and D (P < 0.05). CONCLUSIONS: TGF-ß2 ASON can effectively prevent the proliferation of corneal tissue by inhibiting the activity of TGF-ß2 after injury. The early stage of corneal repair is 7 days after injury, so it is important to use TGF-ß2 ASON at this stage to inhibit the scar hyperplasia. In addition, it is safe to apply TGF-ß2 ASON topically to protect the cornea from obvious side effects.

[Effect of invigorating lung and kidney prescription and its components on secretion of cytokines induced by CSE and LPS].

OBJECTIVE: To observe effect of Invigorating Lung and Kidney Prescription (ILKP) on secretion of cytokines induced by Lipopolysaccharide (LPS) and cigarette smoke extract (CSE) in vitro and discuss prescription regularity of ILKP and reveal molecular mechanism of ILKP treatment on COPD. METHODS: Alveolar epithelial cells A549, airway epithelial cells H292 and monocyte macrophage THP-1 were stimulated with LPS and CSE, in which samples were divided into normal group (20% normal rat serum), model group (20% normal rat serum and 5% CSE with 75 pg/mL LPS), ILKP group and its Components [20% Replenishing qi group (1), Replenishing Lung and Kidney group (2), Activating blood group (3), Reduce phlegm group (4), Replenishing qi with Replenishing Lung and Kidney group (5), Replenishing qi with Activating blood group (6), Replenishing qi with Reduce phlegm group (7), Replenishing qi with Replenishing Lung and Kidney and Activating blood group (8), Replenishing qi with Replenishing Lung and Kidney and Reduce phlegm group (9), Replenishing qi with Activating blood and Reduce phlegm group (10), ILKP serum group (11)]. The cytokines were detected by ELISA and the transcription factors of cytokines were analyzed with Network tool. RESULTS: Compared with the normal group,the secretion of MMP-9, GM-CSF and TGF-beta by THP-1 cells,TIMP by A549 cells,TNF-alpha, MMP-9, GM-CSF, CD36 and TIMP-1 by H292 cells all were increased, while secretion of TNF-alpha, PPAP2B, IL-3 and SOD decreased by THP-1 cells, MMP-9, TNF-alpha, TGF-beta and IL-3 by A549 cells, IL-8 by H292 cells all were decreased. Compared with model group, for THP-1 cells, the secretion of GM-CSF in 3, 6, 10 and 11 prescription groups,TGF-beta in 10 and 11 prescription groups all were decreased. MMP-9 decreased in 9 and 5 prescription groups while it increased in 1, 2, 3, 4, 7, 8, 10 and 11 prescription groups. TNF-alpha was increased in 1, 2, 3, 4, 6, 8, and 9 prescription groups, as it decreased in 11 prescription group. Except 2 prescription group, IL-3 was increased in all other drugs groups. SOD in 2, 4, 5, 8, 9, 11 prescription groups, and PPAP2B in 1, 2, 3, 4, 5, 6, 7 prescription groups were increased while PPAP2B was decreased in 11 prescription group. For A549 cells, the secretion of TIMP-1 was increased in 1, 2, 3, 4, and 5 prescription groups, while it was decreased in 11 prescription groups; Except 1 and 4 prescription group, MMP-9 was increased in other groups; Except TGF-beta was increased in 11 prescription groups, it was increased in other groups; IL-3 was increased in 1, 2, 4, 5, 11 and 3 prescription groups. For H292 cells, the secretion of GM-CSF in 5,6,7 and 2 prescription groups,TNF-alpha in 1, 2, 4, 6, 7, 8, 9 and 10 prescription groups, MMP-9 in 4, 8, 7 and 9 prescription groups all were decreased. CONCLUSION: The effect of ILKP and its components on secretion of cytokines induced by LPS and CSE and its target cells are different, in which Activating blood prescription focuses on inflammation cytokines, Replenishing Lung and Kidney prescription on Lipid metabolism and redox regulation. The effect of ILKPA is the most widely, followed by Activating blood Replenishing Lung and Kidney, and Reduce phlegm prescription on Protease regulation.

[Evaluation of calcium dobesilate for its anti-cataract potential in experimental rat models].

OBJECTIVE: To investigate the preventive and therapeutic effects of agent calcium dobesilate(CDO) with different doses on the galactose cataract of rats. METHODS: We chose fifty Wistar rats at 20- day old. Then, they were divided into 3 groups at random. Choose 10 rats as the control group and gave normal diet; 10 rats as the model group and fed with Gal solution ( drink 12.5% Gal solution from 1 to 7 days and 10%Gal solution from 8 to 21 days except for normal diet ) ; 30 rats as the treatment group and fed with the same Gal solution as the model group, besides they were divided into high dosage group, medium dosage group and low dosage group equally and gave 300 mg×kg(-1)×d(-1), 150 mg×kg(-1)×d(-1), 75 mg×kg(-1)×d(-1) dose of calcium dobesilate respectively from the first day to the end of experiment. The experiment lasts 21 days. Lens opacity were observed and recorded by slit-lamp examination regularly. Activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) and content of malondialdehyde (MDA) were determined to estimate the effect of CDO . Lens fibers changes and Histological changes were observed by scanning electron microscope (SEM) and light microscope (LM) separately. The apoptosis rate of lens epithelium were determined by TUNEL assay. RESULTS: The appearance of Lens opacity in model group was more quickly than that in treatment group in model group, 3 eyes in degree IV, 7 eyes in degree V, while in treatment group, 5 eyes in degree III, 3 eyes in degree IV, 2 eyes in degree V (H = 7.12, P < 0.05). The activity of SOD and GSH-px in treatment group is higher than mode group, but lower than control group on 8th day, there was difference noticed in the activity of SOD (50.01 ± 1.19), (39.39 ± 1.70) , treatment group (46.57 ± 1.09, 46.42 ± 0.87, 45.70 ± 1.46) U/mgProt (F = 88.70, P < 0.05) and the activity of GSH-px (42.92 ± 0.97) , (12.70 ± 1.17) , treatment group (29.16 ± 1.05, 29.08 ± 0.98, 28.25 ± 0.98) nmol/mgprot (F = 1071.89, P < 0.05) ]in 3 groups. The content of MDA in model group (3.43 ± 0.15)nmol/mgprot is higher than treatment group (2.89 ± 0.11, 2.99 ± 0.12, 2.99 ± 0.09)nmol/mgprot (F = 64.62; P < 0.05). There were no statistic significant differences among high dosage group, medium dosage group and low dosage group . The texture of lens fibres detected by SEM in the rats of model was more disorder than treatment group. After HE staining, Lens epithelial cell detected by LM in control group have a clear structure, however, Lens epithelial cell both in model group and treatment group have changed from the initial single layer to multi-storey. Junction between lenses fibers became decreased even disappeared . The apoptosis rate of lens epithelium in treatment group[(2.37 ± 0.17)%, (2.46 ± 0.26)%, (2.79 ± 0.41)%] is higher than control group (0.23 ± 0.07) %, but is much fewer than model group (4.99 ± 0.51) % (χ(2) = 40.41;P < 0.05). CONCLUSION: CDO with different doses could protect lens of rats against galactose damage and there were no significant differences among the different doses of groups.

[Clinical observation of visual quality after aspherical intraocular lens implantation in traumatic cataract].

OBJECTIVE: To observe the visual quality of aspherical intraocular lens (IOL) implantation in traumatic cataract patients. METHODS: Prospective clinical study.96 traumatic cataract patients (96 eyes) suffered from penetrating corneal trauma chosen from the first affiliated hospital of Zhengzhou university during June 2009 to June 2012. They were divided into two groups based on the different type of intraocular lens. The experimental group (48 eyes) was implanted with aspherical IOL and the control group (48 eyes) was implanted with traditional sphere IOL.Uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), contrast sensitivity (CS) and stereoscopic vision were observed at 1 month, 3 months, and 6 months after surgery. At the same time, questionnaire survey about the satisfaction of patients was also performed. The t test was used to compare the preoperative general condition, postoperative visual acuity, contrast sensitivity and stereoscopic vision of the two groups, and the rank sum test was used to compare the astigmatism and the satisfaction of patients. RESULTS: There was no significant difference in UCVA (t = 1.37, 1.28,0.71, P > 0.05) between the experimental group (0.56 ± 0.22, 0.68 ± 0.13,0.84 ± 0.15) and the control group (0.51 ± 0.17, 0.61 ± 0.20,0.81 ± 0.17) at three time points. There was no significant difference in BCVA (t = 0.87, 1.38, 1.39, P > 0.05) between the experimental group (0.62 ± 0.13, 0.74 ± 0.21, 0.87 ± 0.10) and the control group (0.57 ± 0.25,0.69 ± 0.22,0.84 ± 0.15) . The same result happened in stereoscopic vision at 6 months after surgery (far stereopsis:123.5 ± 7.8 vs 126. 9 ± 5.9, t = 0.64, P > 0.05;near stereopsis:90.5 ± 7.8 vs 95.2 ± 3.5; t = 1.36, P > 0.05) between experimental group and control group. The contrast sensitivity of the experimental group in every stage (3 c/d:1.52 ± 0.18, 6 c/d:1.68 ± 0.19, 12 c/d:1.29 ± 0.14, 18 c/d:1.04 ± 0.20) was superior to the control group (3 c/d:1.49 ± 0.27, 6 c/d:1.57 ± 0.21, 12 c/d:1.14 ± 0.20, 18 c/d:0.85 ± 0.14) , especially on the glare sensitivity (the experimental group:3 c/d:1.40 ± 0.15, 6 c/d:1.52 ± 0.22, 12 c/d:1.21 ± 0.18, 18 c/d:0.91 ± 0.14, the control group:3 c/d:1.13 ± 0.13, 6 c/d:1.13 ± 0.28, 12 c/d:0.92 ± 0.13, 18 c/d:0.54 ± 0.16) Compared two groups of difference have statistical significance (free from glare:3 c/d:t = 2.829, 6 c/d:t = 4.092, 12 c/d:t = 3.055, 18 c/d:t = 2.093;glare:3 c/d:t = 2.650, 6 c/d:t = 3.105, 12 c/d:t = 3.395, 18 c/d:t = 2.215;P < 0.05) .Questionnaire survey showed the experimental group (72.9%) was statistically significantly higher (t = 3.016, P < 0.05) than that in the control group (54.1%) on the satisfaction of patients. CONCLUSIONS: The visual quality with implantation of aspherical IOL in traumatic cataract patients is superior to traditional sphere IOL. Aspherical IOL is more appropriate for the patients with small and peripheral corneal scar.It can reduce the visual function damage to minimum caused by trauma.

[Studies of burnt folium nelumbinis in anastalsis and processing].

OBJECTIVE: To research the hemostatic components of burnt Folium Nelumbinis and select a better method of processing. METHOD: The anastalsis of medicinal materials between various collection period and the main astrictive parts of burnt Folium Nelumbinis were reviewed by slide method. The powder was observed by microscopic identification. The content of alkaloid was assayed by HPLC. The new method of processing was selected by the orthogonal experiment. RESULT: The lotus leaf in terminal growth can significantly shorten the clotting time and its carbon is more significant which is consistent with other analysis results. The n-butanol part from burnt Folium Nelumbinis can significantly shorten the clotting time. The optimum processing of the new method is 140 degrees C, 20 min and its product meets relevant regulations. CONCLUSION: In preliminary view, n-butanol part from burnt Folium Nelumbinis is the main astrictive site. It is reasonable to make the lotus leaf in terminal growth into carbon. It is feasible to carbonize by muffle furnace.

Amlodipine inhibits TNF-alpha production and attenuates cardiac dysfunction induced by lipopolysaccharide involving PI3K/Akt pathway.

Calcium channel blockers (CCBs) are widely used in the therapy of cardiovascular diseases. Recent studies have shown that several CCBs exerted distinct anti-inflammatory effect in myocardial dysfunction models. The purpose of the present study was to evaluate therapeutic effect and possible mechanism of action of amlodipine, one of the widely used CCBs, on rat cardiac dysfunction during sepsis induced by lipopolysaccharide (LPS). Pretreatment of the rats with amlodipine (10 or 30 mg/kg, i.v.) delayed the fall of mean arterial blood pressure caused by LPS. Amlodipine also significantly inhibited the elevation of plasma tumor necrosis factor alpha (TNF-alpha) and decreased levels of inducible nitric oxide synthase (iNOS) in response to LPS challenge. To investigate the mechanism of the action of amlodipine, neonatal rat cardiomyocytes were used as a model. Amlodipine concentration-dependently decreased the release of TNF-alpha and iNOS protein expression, and suppressed the degradation and phosphorylation of inhibitor of kappaB-alpha (IkappaB-alpha) in LPS-activated neonatal rat cardiomyocytes. Further studies revealed that amlodipine markedly activated phosphatidylinositiol 3-kinase (PI3K) and Akt, downstream of the PI3K signal cascade. Application of PI3K inhibitors, wortmannin and LY294002 attenuated the depression of TNF-alpha and iNOS expression by amlodipine in LPS-induced cardiomyocytes. These findings may explain some cardioprotective effects of amlodipine in LPS-mediated sepsis and suggest that the inhibition of TNF-alpha and iNOS expression by amlodipine is, at least in part, dependent on PI3K/Akt signaling pathway.

[Multiresidue analysis of organochlorine pesticides and the application to Chinese herbal medicine].

[A review of study on multiresidue analyses of organochlorine pesticides(OCPs) was presented here, which included sample extraction, clean-up and determination. The multiresidue analytical methods applied to Chinese herbal medicine was also discussed.

PC-407 inhibited proliferation and induced apoptosis in human colon cancer SW-1116 cells.

AIM: To study whether PC-407 [4-[5-naphthyl-3- (trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide] inhibits cell viability and induces apoptosis in human colon cancer SW-1116 cells. METHODS: Inhibition of SW-1116 proliferation was measured by MTT assay. Morphological assessment of apoptosis was performed with fluorescence microscope and electron microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry. RESULTS: PC-407 inhibited SW-1116 cell proliferation in a concentration-dependent manner after 3 d of treatment, and the IC(50) for PC-407 inhibition of cell number was 16.67+/-0.17 micromol/L. After incubation of SW-1116 cells with PC-407 20 micromol/L for 24 h, morphological changes of typical apoptosis were observed by AO/EB staining or transmission electron microscopy. Flow cytometry analysis showed that PC-407 induced apoptosis in SW-1116 cells in a time- and concentration-dependent manner. The agarose gel electrophoresis of DNA revealed a ladder pattern 48 h later. CONCLUSION: PC-407 inhibited proliferation and induced apoptosis in the human colon cancer SW-1116 cell line.