DNA Extraction from Formalin-fixed and Paraffin-embedded Tissues by Triton X-100 for Effective Amplification of EGFR Gene by Polymerase Chain Reaction

For first-line non-small-cell lung cancer（NSCLC） therapy,detecting mutation status of the epidermal growth factor receptor（EGFR） gene constitutes a prudent test to identify patients who are most likely to benefit from EGFR-tyrosine kinase inhibitor（TKI） therapy.Now,the material for detecting EGFR gene mutation status mainly comes from formalin-fixed and paraffin-embedded（FFPE） tissues.DNA extraction from FFPE and the amplification of EGFR gene by polymerase chain reaction（PCR） are two key steps for detecting EGFR gene mutation.We showed a simple method of DNA extraction from FFPE tissues for the effective amplification of EGFR gene.Extracting DNA from the FFPE tissues of NSCLC patients with 1% Triton X-100（pH=10.0） was performed by heating at 95 °C for 30 min.Meanwhile,a commercial kit was used to extract DNA from the same FFPE tissues of NSCLC patients for comparison.DNA extracted products were used as template for amplifying the exons 18,19,20 and 21 of EGFR by PCR for different amplified fragments.Results show that DNA fragment size extracted from FFPE tissues with 1% Triton X was about 250―500 base pairs（bp）.However,DNA fragment size extracted from FFPE tissues via commercial kit was about from several hundreds to several thousands bp.The DNA yield extracted from FFPE tissues with 1% Triton X was larger than that via commercial kit.For about 500 bp fragment,four exons of EGFR could not be amplified more efficiently from extracted DNA with 1% Triton X than with commercial kit.However,for about 200 bp fragment.This simple and non-laborious protocol could successfully be used to extract DNA from FFPE tissue for the amplification of EGFR gene by PCR,further screening of EGFR gene mutation and facilitating the molecular analysis of a large number of FFPE tissues from NSCLC patients.

Engineering vascularized organotypic tissues via module assembly

Adequate vascularization is a critical determinant for the successful construction and clinical implementation of complex organotypic tissue models. Currently, low cell and vessel density and insufficient vascular maturation make vascularized organotypic tissue construction difficult,greatly limiting its use in tissue engineering and regenerative medicine. To address these limitations, recent studies have adopted pre-vascularized microtissue assembly for the rapid generation of functional tissue analogs with dense vascular networks and high cell density. In this article, we summarize the development of module assembly-based vascularized organotypic tissue construction and its application in tissue repair and regeneration, organ-scale tissue biomanufacturing, as well as advanced tissue modeling.

Insect Resistance of Different Tissues of Transgenic Cotton to Spodoptera exigua(Hbner)

[Objective] The aim was to learn the resistance of different tissues and organs of transgenic cotton to Spodoptera exigua （Hbner）. [Method] Flowers,the 1st,the 3rd,the 6th and the 14th leaves from the top of 33B,GK12 and SGK321 were used to feed S. exigua neonates respectively. Survival larvae and dead ones were counted on the 3rd,the 7th,the 10th,the 16th and the 19th day; meanwhile,the pupae amount was recorded,and the pupae weight was measured at the 24th h after pupation. [Result] The survival curves,pupation rates and pupae weights of S. exigua feeding on different tissues of transgenic cotton were not significantly different from those of S. exigua feeding on the corresponding tissues of conventional cotton; pupation rate of S. exigua feeding on different leaves of the same cotton variety were not significantly different from each other,but all higher than that of S. exigua feeding on the flowers of that cotton; and there were no differences among pupation weights of S. exigua feeding on different leaves or flowers of the same cotton variety. [Conclusion] Transgenic cotton showed weak resistance to S. exigua. Hence,in the transgenic cotton fields,more attention should be paid to occurrence trend of S. exigua and its control.

Expression of Ferric Chelate Reductase Gene in Citrus junos and Poncirus trifoliataTissues

It has been hypothesized that under iron stress high ferric chelate reductase (FCR) activity in the absorptive root of plants tolerant to iron_deficiency will be induced and result in subsequent Fe 2+ transport across the plasmalemma. The activity of FCR and expression of FCR gene (FRO2) in Citrus junos Sieb. ex Tanaka tolerant to iron_deficiency and Poncirus trifoliata (L.) Raf. susceptible to iron_deficiency were determined to elucidate the physiological difference which causes the different tolerance of the two citrus rootstocks to iron stress. The activity of FCR was detectable in excised roots and was stimulated about 20_times in C. junos and only about 3_times in P. trifoliata under iron deficiency for four weeks. The FRO2 of Arabidopsis was used as a probe, the tissue print technique was used to ascertain the expression of the FCR gene in C. junos and P. trifoliata under iron stress. High_level transcripts were observed in the absorptive root, young green stem as well as new leaf of C. junos under iron stress for two weeks, and the transcripts were accumulated only slightly in P. trifoliata at the same time. The results showed that the obvious increase of FCR activity was an important reason for the tolerance of C. junos to iron_deficiency, and the regulation of FCR activity seemed to be at the transcriptional level, and the expression of FRO2 occurred in the root, stem and leaf.

Effects of Weak Light on the Ultrastructural Variations of Phloem Tissues in Source Leaves of Three-Year-Old Nectarine Trees(Prunus persica L.var. nectarina Ait.)

Leaves from three_year_old solar greenhouse nectarine trees ( Prunus persica L. var. nectarina Ait. “Zao Hong Yan”) were used as materials in this study. It was the first time that the ultrastructural characteristics of phloem tissues of source leaves were observed and compared in normal and weak light intensities using the transmission electron microscopy. Results showed that the average diameters of companion cells (CC) and sieve elements (SE) of all kinds of veins were bigger in normal than that in weak light intensity, indicating that light could influence the cell development and growth. Dense cytoplasm with abundant mitochondria, endoplasmic reticulums, multivesicular bodies, vesicles and plastids were observed in normal light intensity. On the contrary, CC with small vacuolar structures and few mitochondrias, endoplasmic reticulums were shown in weak light. Misalignment of grana thylakoid margins of nectarine leaves also was seen in weak light. The sieve pores of SEs were obstructed in weak light. Chloroplasts with numerous starch grains and few mitochondrias were noticed in the mesophyll cell (MES) surrounding the bundle sheath in weak light. The storage of starch grains appeared to result from an unbalance between photosynthate production and export of photosynthates. This observation provided a strong support to the point that most leaves export the most of assimilates in the light time. Plasmodesmal densities between SE/CC, CC/PP (phloem parenchyma cell), PP/PP and PP/BSC (bundle_sheath cell) decreased in weak light. Plasmodesmata were observed between CC/SE (NS) (nacreous_walled sieve element), PP/BSC in branch veins in normal light intensity, but not in weak light. Thus apoplasmic pathway may be the main mode of transport of assimilates in weak light, however symplasmic pathway may be the main mode of transport of assimilates in normal light intensity. These results demonstrated that the solar greenhouse nectarine trees could be adapted to the weak light via the ultrastructure variation of phloem tissues of the source leaves.

Comparison of microRNA profiles of human periodontal diseased and healthy gingival tissues

MicroRNAs （miRNAs） have been clemonstrated to play an important role in regulation of the immunoinflammatory response; however, the function of miRNAs in periodontal inflammation has not been investigated. The objective of this study was to explore the properties of miRNAs in periodontal inflammation by comparing miRNA profiles of inflamed and healthy gingival tissues. Gingival tissues were obtained from 10 periodontitis patients and 10 healthy subjects. After RNA extraction, miRNA profiles were analyzed by microarray, and expression levels of selected miRNAs were confirmed by real-time quantitative reverse transcription polymerase chain reaction （RT-PCR）. Analyses using two computational methods, Targetscan and MicroRNA.org, were combined to identify common targets of these miRNAs. Finally, the individual miRNA expression levels of three toll-like receptor （TLR）-related miRNAs from inflamed and healthy gingival tissues were evaluated by RT-PCR. Ninety-one miRNAs were found to be upregulated and thirty-four downregulated over two-fold in inflamed gingival tissue compared with those in healthy gingival tissue. Twelve selected inflammatory-related miRNAs, hsa-miR-126＊, hsa-miR-20a, hsa-miR-142-3p, hsa-miR-19a, hsa-let-7f, hsa-miR-203, hsa-miR-17, hsa-miR-223, hsa-miR-146b, hsa-miR-146a, hsa-miR-155, and hsa-miR-205 showed comparable expression levels by microarray and real-time quantitative RT-PCR analyses. In addition, the putative inflammation targets of these miRNAs were predicted, and three that were tested （hsa-miRNA-146a, hsa-miRNA-146b, and hsa-miRNA-155）, showed significant differences between inflamed and healthy gingiva. This remarkable difference in miRNA profiles between periodontal diseased and healthy gingiva implicates a probable close relationship between miRNAs and periodontal inflammation. The data also suggest that the regulation of TLRs in periodontal inflammation may involve miRNA pathways.

3D Bioprinting:A Novel Avenue for Manufacturing Tissues and Organs

Three-dimensional(3D)bioprinting is a rapidly growing technology that has been widely used in tissue engineering,disease studies,and drug screening.It provides the unprecedented capacity of depositing various types of biomaterials,cells,and biomolecules in a layer-by-layer fashion,with precisely controlled spatial distribution.This technology is expected to address the organ-shortage issue in the future.In this review,we first introduce three categories of 3D bioprinting strategies:inkjet-based printing(IBP),extrusion-based printing(EBP),and light-based printing(LBP).Biomaterials and cells,which are normally referred to as“bioinks,”are then discussed.We also systematically describe the recent advancements of 3D bioprinting in fabricating cell-laden artificial tissues and organs with solid or hollow structures,including cartilage,bone,skin,muscle,vascular network,and so on.The development of organs-onchips utilizing 3D bioprinting technology for drug discovery and toxicity testing is reviewed as well.Finally,the main challenges in current studies and an outlook of the future research of 3D bioprinting are discussed.

Androgen receptors are expressed in a variety of human fetal extragenital tissues： an immunohistochemical study

Aim： To investigate the expression of androgen receptors in the extragenital tissues of developing human embryo. Methods： Using immunohistochemistry, we investigated the distribution of androgen receptor （AR） in the extragenital tissues of paraffin-embedded tissue sections of first trimester （8-12 weeks gestation） human embryos. Gender was determined by polymerized chain reaction. Results： There were no differences in the expression and distribution of AR in male and female embryos at any stage of gestation. AR expression was seen in the thymus gland. The bronchial epithelium of the lungs showed intense positive staining with surrounding stroma negative. Furthermore, positive staining for androgen receptor was exhibited in the spinal cord with a few positive cells in the surrounding tissues. Cardiac valves also showed strong positive staining but with faint reactivity of the surrounding cardiac muscle. There was no staining in kidney, adrenal, liver or bowel. Conclusion： This study demonstrates that immunoreactive AR protein is present in a wide variety of human first trimester fetal tissues and shows the potential for androgen affecting tissues, which are mostly not considered to be androgen dependent. Moreover, it implies that androgen might act as atrophic factor and affect the early development of these organs rather than simply sexual differentiation.

Immunomodulatory effects of mesenchymal stem cells derived from adipose tissues in a rat orthotopic liver transplantation model

BACKGROUND: Acute rejection after liver transplantation is usually treated with large doses of immunosuppressants with severe toxic and side-effects, so it is imperative to find a safe and effective method for preventing and treating rejection. This study was designed to confirm the immunomodulatory effects of rat mesenchymal stem cells (MSCs) in vitro and investigate the tolerogenic features in a rat model of allogeneic liver transplantation. METHODS: MSCs were isolated from adipose tissue of Sprague-Dawley (SD) rats and cultured. In vitro, MSCs were added into a mixed lymphocyte culture (MLC) system to study the inhibitory effects of MSCs on the proliferation of T lymphocytes in Wistar rats. By using SD and Wistar rats as liver donors and recipients, an orthotopic liver transplantation model was established and the rats were divided into a MSC-treated group and a blank control group. On postoperative day 7, all rats were sacrificed, and the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), interleukin-2 (IL-2) and interleukin-10 (IL-10) were measured. The pathological changes of liver tissue and apoptosis of hepatocytes were also assessed. RESULTS: In in vitro MLC, T lymphocyte proliferation in Wistar rats was significantly inhibited by 48.44%. In the MSC-treated group, the levels of ALT, AST, TBIL, IL-2 and IL-10 were 134.2 +/- 45.0 U/L, 162.5 +/- 30.5 U/L, 30.6 +/- 5.4 mu mol/L, 187.35 +/- 18.26 mu g/L and 193.95 +/- 37.62 mu g/L, and those in the blank control group were 355.6 +/- 54.3 U/L, 296.4 +/- 71.2 U/L, 145.7 +/- 28.6 +/- mol/L, 295.73 +/- 57.15 mu g/L and 75.12 +/- 11.23 mu g/L, respectively, with statistically significant differences (P<0.05). Pathological examination revealed that the rejection in the MSC-treated group was clearly alleviated compared with that in the blank control group. TUNEL indicated that the apoptosis of hepatocytes in the MSC-treated group was milder than that in the blank control group (P<0.05). CONCLUSION: Adipose-derived MSCs clearly inhibit recipient-derived T lymphocyte proliferation in MLC and significantly alleviate acute rejection following orthotopic liver transplantation in rats.

Phenolic and flavonoid contents of mandarin (Citrus reticulata Blanco) fruit tissues and their antioxidant capacity as evaluated by DPPH and ABTS methods

The total phenolic and flavonoid contents in the fruit tissues （peels, pulp residues, seeds, and juices） of 19 citrus genotypes belonged to Citrus reticulata Blanco were evaluated and their antioxidant capacity was tested by 2,2-diphenyl-l-picrylhydra- zyl radicals （DPPH） method and 2,2＇-azino-bis（3-ethylbenzthiozoline-6）-sulphonic acid （ABTS） method. The total phenolic and flavonoid contents, and their antioxidant capacity varied in different citrus fruit tissues. Generally, the peel had both the highest average of total phenolics （27.18 mg gallic acid equivalent （GAE） g^-1 DW） and total flavonoids （38.97 mg rutin equivalent （RE） g^-1 DW）. The highest antioxidant capacity was also the average of DPPH value （21.92 mg vitamin C equiv- alent antioxidant capacity （VCEAC） g^-1 DW） and average of ABTS value （78.70 mg VCEAC g-1 DW） in peel. The correlation coefficient between the total phenolics and their antioxidant capacity of different citrus fruits tissues ranged from 0.079 to 0.792, and from -0.150 to 0.664 for the total flavonoids. The antioxidant capacity of fruit tissues were correlated with the total phenoilc content and flavonoid content except in case of the peel. In addition, the total phenolic content and antioxidant capacity varied in different citrus genotypes. Manju and Karamandarin were better genotypes with higher antioxidation and the phenolic content, however Shagan was the poorest genotype with lower antioxidation and the phenolic content.

Role of nanotopography in the development of tissue engineered 3D organs and tissues using mesenchymal stem cells

Recent regenerative medicine and tissue engineering strategies(using cells, scaffolds, medical devices and gene therapy) have led to fascinating progress of translation of basic research towards clinical applications. In the past decade, great deal of research has focused on developing various three dimensional(3D) organs, such as bone, skin, liver, kidney and ear,using such strategies in order to replace or regenerate damaged organs for the purpose of maintaining or restoring organs' functions that may have been lost due to aging, accident or disease. The surface properties of a material or a device are key aspects in determining the success of the implant in biomedicine, as the majority of biological reactions in human body occur on surfaces or interfaces. Furthermore, it has been established in the literature that cell adhesion and proliferation are, to a great extent, influenced by the micro- and nanosurface characteristics of biomaterials and devices. In addition, it has been shown that the functions of stem cells, mesenchymal stem cells in particular, could be regulated through physical interaction with specific nanotopographical cues. Therefore, guided stem cell proliferation, differentiation and function are of great importance in the regeneration of 3D tissues and organs using tissue engineering strategies. This review will provide an update on the impact of nanotopography on mesenchymal stem cells for the purpose of developing laboratory-based 3D organs and tissues, as well as the most recent research and case studies on this topic.

Development of An Enzyme-Linked Immunosorbent Assay for Determination of the Furaltadone Etabolite,3-Amino-5-Morpholinomethyl-2-Oxazolidinone(AMOZ) in Animal Tissues

Objective To determine 3-amino-5-morpholinomethyl-2-oxazolidinone （AMOZ） residues released from protein bound AMOZ in animal tissues. Methods Polyclonal and monoclonal antibodies were produced in this study. A rapid, sensitive, and specific competitive direct enzyme-linked immunosorbent assay （cdELISA） was developed. Results Rabbit polyclonal antibodies were used in the optimized cdELISA method, and exhibited negligible cross-reactivity with other compounds structurally related to AMOZ. The IC50 of the polycional antibody was 0.16 ng/mL The method limit of detection in four different types of animal and fish tissues was less than 0.06 μg/kg. Recoveries ranged from 80% to 220% for fortified samples with the coefficient of variation values less than 15%. The results of the cdELISA method were in good agreement with the results from an established liquid chromatography-tandem mass spectrometry confirmatory method used for AMOZ residues. Conclusion The cdELISA method developed in the present study is a convenient practical tool for screening large numbers of animal and fish tissue samples for the the detection of released protein bound AMOZ residues.

Dedifferentiated fat cells: an alternative source of adult multipotent cells from the adipose tissues

When adipose-derived stem cells （ASCs） arc retrieved from the stromal vascular portion of adipose tissue, a large amount of mature adipocytes are often discarded. However, by modified ceiling culture technique based on their buoyancy, mature adipocytes can be easily isolated from the adipose cell suspension and dediffercn- tiated into lipid-frce fibroblast-like cells, named dediffercntiated fat （DFAT） cells. DFAT cells rc-establish active proliferation ability and undertake multipotent capacities. Compared with ASCs and other adult stem cells, DFAT cells showed unique advantages in their abundance, isolation and homogeneity. In this concise review, the establishment and culture methods of DFAT cells arc introduced and the current profiles of their cellular nature are summarized. Under proper inducti~,n culture in vitro or environment in vivo, DFAT cells could demonstrate adipogenic, osteogenic, chondrogenie and myogenic potentials. In angiogenie conditions, DFAT cells could exhibit perivascular characteristics antt elicit neovascularization. Our preliminary findings also suggested the pericyte phenotype underlying such cell lineage, which supported a novel interpretation about the common origin of mesenchymal stem cells and tissue-specific stem cells within blood vessel walls. Current research on DFAT cells indicated that this alternative source of adult multipotent cells has great potential in tissue engineering and regenerative medicine.

Prepartum body conditions affect insulin signaling pathways in postpartum adipose tissues in transition dairy cows

Background: Overconditioned dairy cows are susceptible to excessive lipolysis and increased insulin resistance during the transition period.The associations among body fat reserve,insulin resistance,and lipolysis in adipose tissues(AT) remain to be elucidated.Therefore,this study aimed to investigate whether excessive fat reserves influence the insulin signaling pathway in AT postpartum.Results: Twenty multiparous dairy cows were selected and assigned to one of two groups,according to prepartum body condition score(BCS): Control group(BCS = 3.0–3.5;n = 10) and Overconditioned group(BCS ≥ 4.0;n = 10).Blood samples were collected on days-14,-7,-4,-2,-1,0,1,2,4,7,and 14 relative to parturition.Subcutaneous AT were collected on day 2 following parturition for quantitative real-time polymerase chain reaction and western blot analyses.No differences were observed between the two groups in serum glucose,non-esterified fatty acids,β-hydroxybutyric acid,tumor necrosis factor(TNF)α,insulin,or leptin concentrations during the experimental period.Compared with the control cows,the overconditioned cows had lower serum triglyceride levels and higher adiponectin concentrations.In the AT postpartum,insulin receptor mRNA and protein levels were lower in the overconditioned cows than in the control cows,and no differences were found in glucose transporter 4 mRNA.Compared with the control cows,the overconditioned cows had lower mRNA levels of TNFα and higher mRNA levels of peroxisome proliferator-activated receptor gamma(PPARγ) in AT postpartum.The phosphorylated protein kinase B(AKT) content and phosphorylation rate of AKT were increased in the overconditioned cows compared with the control cows,which suggested that the downstream insulin signaling in AT was affected.Conclusions: In the present study,transition dairy cows with higher BCS did not show more fat mobilization.The changes of insulin signaling pathway in AT postpartum of overconditioned cows may be partly related to the expression of PPARγ and TNFα,and the secretion of adiponectin.

A dynamic cellular vertex model of growing epithelial tissues

Intercellular interactions play a significant role in a wide range of biological functions and processes at both the cellular and tissue scales, for example, embryogenesis, organogenesis, and cancer invasion. In this paper, a dynamic cellular vertex model is presented to study the morphomechanics of a growing epithelial monolayer. The regulating role of stresses in soft tissue growth is revealed. It is found that the cells originating from the same parent cell in the monolayer can orchestrate into clustering patterns as the tissue grows. Collective cell migration exhibits a feature of spatial correlation across multiple cells. Dynamic intercellular interactions can engender a variety of distinct tissue behaviors in a social context. Uniform cell proliferation may render high and heterogeneous residual compressive stresses, while stress-regulated proliferation can effectively release the stresses, reducing the stress heterogeneity in the tissue. The results highlight the critical role of mechanical factors in the growth and morphogenesis of epithelial tissues and help understand the development and invasion of epithelial tumors.

Photon attenuation parameters for some tissues from Geant4simulation, theoretical calculations and experimental data:a comparative study

Mass attenuation coefficients, effective atomic numbers, effective electron densities and Kerma relative to air for adipose, muscle and bone tissues have been investigated in the photon energy region from 20 keV up to 50 MeV with Geant4 simulation package and theoretical calculations. Based on Geant4 results of the mass attenuation coefficients, the effective atomic numbers for the tissue models have been calculated. The calculation results have been compared with the values of the Auto-Zeff program and with other studies available in the literature. Moreover, Kerma of studied tissues relative to air has been determined and found to be dependent on the absorption edges of the tissue constituent elements.

Nuclear magnetic resonance-based metabolomics and metabolic pathway networks from patient-matched esophageal carcinoma,adjacent noncancerous tissues and urine

BACKGROUND Several studies have demonstrated a correlation between esophageal cancer(EC)and perturbed urinary metabolomic profiles,but none has described the correlation between urine metabolite profiles and those of the tumor and adjacent esophageal mucosa in the same patient.AIM To investigate how urinary metabolic phenotypes were linked to the changes in the biochemical landscape of esophageal tumors.METHODS Nuclear magnetic resonance-based metabolomics were applied to esophageal tumor tissues and adjacent normal mucosal tissues alongside patient-matched urine samples.RESULTS Analysis revealed that specific metabolite changes overlapped across both metrics,including glucose,glutamate,citrate,glycine,creatinine and taurine,indicating that the networks for metabolic pathway perturbations in EC,potentially involved in but not limited to disruption of fatty acid metabolism,glucose and glycolytic metabolism,tricarboxylic acid cycle and glutaminolysis.Additionally,changes in most urinary biomarkers correlated with changes in biomarker candidates in EC tissues,implying enhanced energy production for rapid cell proliferation.CONCLUSION Overall,these associations provide evidence for distinct metabolic signatures and pathway disturbances between the tumor tissues and urine of EC patients,and changes in urinary metabolic signature could reflect reprogramming of the aforementioned metabolic pathways in EC tissues.Further investigation is needed to validate these initial findings using larger samples and to establish the underlying mechanism of EC progression.

Mesenchymal stromal cells from human perinatal tissues: From biology to cell therapy

Cell-based regenerative medicine is of growing interest in biomedical research. The role of stem cells in this context is under intense scrutiny and may help to define principles of organ regeneration and develop innovative therapeutics for organ failure. Utilizing stem and progenitor cells for organ replacement has been conducted for many years when performing hematopoietic stem cell transplantation. Since the first successful transplantation of umbilical cord blood to treat hematological malignancies, non-hematopoietic stem and progenitor cell populations have recently been identified within umbilical cord blood and other perinatal and fetal tissues. A cell population entitled mesenchymal stromal cells (MSCs) emerged as one of the most intensely studied as it subsumes a variety of capacities: MSCs can differentiate into various subtypes of the mesodermal lineage, they secrete a large array of trophic factors suitable of recruiting endogenous repair processes and they are immunomodulatory.Focusing on perinatal tissues to isolate MSCs, we will discuss some of the challenges associated with these cell types concentrating on concepts of isolation and expansion, the comparison with cells derived from other tissue sources, regarding phenotype and differentiation capacity and finally their therapeutic potential.

Reaction of antibodies to Campylobacter jejuni and cytolethal distending toxin B with tissues and food antigens

BACKGROUND The bacteria Campylobacter jejuni(C. jejuni) is commonly associated with GuillaneBarré syndrome(GBS) and irritable bowel syndrome(IBS), but studies have also linked it with Miller Fisher syndrome, reactive arthritis and other disorders, some of which are autoimmune. It is possible that C. jejuni and its toxins may be crossreactive with some human tissues and food antigens, potentially leading to autoimmune responses.AIM To measure the immune reactivity of C. jejuni and C. jejuni cytolethal distending toxin(Cdt) antibodies with tissue and food antigens to examine their role in autoimmunities.METHODS Using enzyme-linked immunosorbent assay(ELISA) methodology, specific antibodies made against C. jejuni and C. jejuni Cdt were applied to a variety of microwell plates coated with 45 tissues and 180 food antigens. The resulting immunoreactivities were compared to reactions with control wells coated with human serum albumin(HSA) which were used as negative controls and with wells coated with C. jejuni lysate or C. jejuni Cdt which served as positive controls.RESULTS At 3 SD above the mean of control wells coated with HSA or 0.41 OD, the mouse monoclonal antibody made against C. jejuni showed moderate to high reactions with zonulin, somatotropin, acetylcholine receptor, β-amyloid and presenilin.This immune reaction was low with an additional 25 tissue antigens including asialoganglioside, and the same antibody did not react at all with another 15 tissue antigens. Examining the reaction between C. jejuni antibody and 180 food antigens, we found insignificant reactions with 163 foods but low to high immune reactions with 17 food antigens. Similarly, we examined the reaction of C. jejuni Cdt with the same tissues and food antigens. The strongest reactions were observed with zonulin, intrinsic factor and somatotropin. The reaction was moderate with 9 different tissue antigens including thyroid peroxidase, and reaction was low with another 10 different antigens, including neuronal antigens.The reaction of C. jejuni Cdt antibody with an additional 23 tissue antigens was insignificant. Regarding the reaction of C. jejuni Cdt antibody with different food antigens, 160 out of 180 foods showed insignificant reactions, while 20 foods showed reactions ranging from low to high.CONCLUSION Our findings indicate that C. jejuni and its Cdt may play a role in inflammation and autoimmunities beyond the gut.

Repair cell first,then regenerate the tissues and organs

Wound healing,tissue repair and regenerative medicine are in great demand,and great achievements in these fields have been made.The traditional strategy of tissue repair and regeneration has focused on the level of tissues and organs directly;however,the basic process of repair at the cell level is often neglected.Because the cell is the basic unit of organism structure and function;cell damage is caused first by ischemia or ischemia-reperfusion after severe trauma and injury.Then,damage to tissues and organs occurs with massive cell damage,apoptosis and even cell death.Thus,how to achieve the aim of perfect repair and regeneration?The basic process of tissue or organ repair and regeneration should involve repair of cells first,then tissues and organs.In this manuscript,it is my consideration about how to repair the cell first,then regenerate the tissues and organs.