Preconditioning effects on expression of proto-oncogenes c-fos and c-jun after hepatic ischemia/reperfusion in rats

BACKGROUND: Ischemia/reperfusion is the main cause of hepatic damage in liver transplantation. Immediate early genes (IEGs) encode proteins can regulate expression of cellular response genes after injury, and is associated with tissue repair and cell apoptosis. The purpose of this re- search was to investigate the effects of preconditioning on expression of immediate early genes c-fos and c-jun follow- ing hepatic ischemia/reperfusion (IR) and its roles in cellu- lar regeneration and apoptosis. METHODS: Ninety-six Wistar rats were randomly divided into IR group and hepatic ischemic preconditioning (IPC) group, and each group was further divided into eight sub- groups (n =6). The model of partial liver ischemia/reper- fusion was used. The rats were subjected to 60-minute liver ischemia, preceded by 10-minute preconditioning. After 0-, 0.5-, 1-, 2-, 4-, 8-, 12-, 24-hour reperfusion, the se- rum and liver tissue in each group were collected to detect the level of serum ALT/AST, liver histopathology, expres- sion of c-fos, and c-jun mRNA. Flow cytometer was used to detect Ki67 and Sub-G1 as the quantity indicators of cell regeneration and apoptosis respectively. RESULTS: Compared with IR group, IPC group showed a significantly lower ALT/AST level in 0. 5-hour sub-group to 8-hour sub-group (P<0.05). Ki67 elevated significantly at 0.5, 1, 2 hours, but decreased significantly at 24 hours ( P < 0 . 05). Ap index decreased significantly after 1-hour reperfusion(P<0.05). Expressions of c-fos and c-jun mR- NA were low, especially c-jun at 0.5, 1 and 2 hours after reperfusion. CONCLUSION: Ischemic preconditioning can protect liver cells against ischemia/reperfusion injury, and this protec- tive effect may be related to influence transcription levels of c-fos and c-jun.

Effect of Antisense Oligodeoxynucleotide of Vascular Endothelial Growth Factor C on Lymphangiogenesis and Angiogenesis of Pancreatic Cancer

In order to investigate the effect of antisense oligonucleotide （ASODN） of vascular endothelial growth factor C （VEGF-C） on lymphangiogenesis and angiogenesis of pancreatic cancer, antisense and scamble-sense oligonucleotide of VEGF-C were constructed, and the model of nude mice with orthotopically xenografied human pancreatic cancer cells （Panc-1） was established. Thirty nude mice were randomly divided into 3 groups： PBS control group （group A）, scramble-sense control group （group B） and antisense group （group C）. All nude mice were treated once every 2 days as 3 times per week, for 3 weeks （oligonucleotide 10 mg/kg every time）. After treatments were completed, ELISA method was used to examine the concentration of VEGF-C in plasma and immunohistochemical method to examine microvessel density （MVD）, lymphtic vessel density （LVD） of pancreatic cancer. The results showed that the expression of VEGF-C was inhibited significantly in group C. The concentrations were 237.5±41.5, 221.5±52.3 and 108.6±14.9 pg/mL in groups A, B and C respectively （P〈0.01）. LVD in groups A, B and C was 13.8±2.1, 12.4±1.9 and 4.2±1.6 respectively （P〈0.01）. MVD in groups A, B and C was 27.5±8.7, 25.9±4.2 and 19.4±5.6 respectively with no significant difference among the groups （P〉0.05）. It was suggested that VEGF-C ASODN decreased the expression levels of VEGF-C in nude mice with orthotopically xenografted human pancreatic cancer, and it could inhibit lymphangiogenesis, but had no significant effect on angiogenesis.

The expression of c-src gene in the carcinogenesis process of human cardia adenocarcinoma

AIM To investigate the activation, expression of c src gene and its role in the carcinogenetic process of human cardia adenocarcinoma (CA). METHODS Fifty six cases of CA, 34 cases of normal, 36 cases of protiferative epithelia adjacent to carcinoma, and 20 cases of lymph node metastases of CA were studied for PP60 c src , the expression product of c src gene immunohistochemically by using the specific monoclonal antibody, Mab327. RESULTS The positive rates of PP60 c src in the normal epithelia, protiferative epithelia, CA and lymph node metastases were 29 4% (10/*!34), 94 4% (34/*!36), 71 4% (40/*!56) and 60 0% (12/*!20) , respectively, among them, the differences of the positive rates were statistically significant ( P <0 01) . The expression levels of PP60 c src in CA and proliferative epithelia were significantly higher than that in the normal epithelia ( P <0 01) . The PP60 c src positive rates in the papillary, tubular, poorly differentiated and mucous adenocarcinoma were 75 0% (6/*!8) , 81 8% (18/*!22) , 50 0% (10/*!20) and 100 0% (6/*!6) , respectively, whereas those of tubular and mucous adenocarcinomas were significantly higher than those of papillary and poorly differentiated adenocarcinomas ( P <0 05) , and the PP60 c src expression levels of tubular and mucous adenocarcinomas were also significantly higher than those of papillary and poorly differentiated adenocarcinomas ( P <0 01) . CONCLUSION The activation and expression of c src gene are associated with the initiation and development of human CA; the protein amount of PP60 c src increased during the process of carcinogenesis; and PP60 c src expression is also related to lymph node metastases.

Identification of key genes involved in axon regeneration and Wallerian degeneration by weighted gene co-expression network analysis

Peripheral nerve injury repair requires a certain degree of cooperation between axon regeneration and Wallerian degeneration.Therefore,investigating how axon regeneration and degeneration work together to repair peripheral nerve injury may uncover the molecular mechanisms and signal cascades underlying peripheral nerve repair and provide potential strategies for improving the low axon regeneration capacity of the central nervous system.In this study,we applied weighted gene co-expression network analysis to identify differentially expressed genes in proximal and distal sciatic nerve segments from rats with sciatic nerve injury.We identified 31 and 15 co-expression modules from the proximal and distal sciatic nerve segments,respectively.Functional enrichment analysis revealed that the differentially expressed genes in proximal modules promoted regeneration,while the differentially expressed genes in distal modules promoted neurodegeneration.Next,we constructed hub gene networks for selected modules and identified a key hub gene,Kif22,which was up-regulated in both nerve segments.In vitro experiments confirmed that Kif22 knockdown inhibited proliferation and migration of Schwann cells by modulating the activity of the extracellular signal-regulated kinase signaling pathway.Collectively,our findings provide a comparative framework of gene modules that are co-expressed in injured proximal and distal sciatic nerve segments,and identify Kif22 as a potential therapeutic target for promoting peripheral nerve injury repair via Schwann cell proliferation and migration.All animal experiments were approved by the Institutional Animal Ethics Committee of Nantong University,China(approval No.S20210322-008)on March 22,2021.

Mapping Major Restore Genes for C-type Cytoplasmic Male Sterility in Maize with SSR Marker

Through observation about the restoration of male fertility of F2 and BC, progeny, we found that the restoring line Fengkel had two duplicating restorer genes. The restorer gene R/5 in Fengkel background was located on chromosome 5L by SSR method; it linked with bnlg1711, bnlg1346 and phi058, the genetic distances with bnlg1711, bnlg1346, and phi058 were 7.51cM, 1.68cM, and 9.87cM respectively; the restorer gene Rf4 was mapped on chromosome 8S linked with bnlg2307.

Ultraviolet-attenuated cercariae ofSchistosoma japonicum fail to effectively induce a Th1 response in spite of up-regulating expression of cytotoxicity-related genes in C57BL/6 mice

Objective： To better understand the reason that Schistosoma japonicurn （S. japonicum） ultraviolet （UV）- radiated cercariae could not induce high level of protection in C57BL/6 mice. Methods： Microarray technology was performed to investigate the gene transcription profile in skin draining lymph nodes （sdLNs） at 1 w after exposure to attenuated cercariae （AC） or normal cercariae （NC） of S. japonicum in C57BL/6 mice. The expressions of some representative genes were further confirmed by real-time PCR. Subsequently, the expressions of Th1/Th2 cytokine genes, cytotoxicity-related genes, as well as co-stimulator genes in spleens from AC-vaccinated and NC- infected mice were analyzed by real-time PCR at w 3 and 6 post-exposure. Results： The gene expressions of Th1 cytokines, including interferon-y （IFN-γ）, interleukin （IL）-12 and tumor necrosis factor-α （TNF-α） in the sdLNs were significantly lower in AC-vaccinated mice than in NC-infected mice. Furthermore, the gene expressions of Th1- and Th2- cytokines, including IFN-γ, IL-12, TNF-α, IL-4 and IL-10, in the spleens from AC-vaccinated mice showed little changes at w 3 and 6 post-vaccination. In addition, cytotoxicity-related molecules including granzyme A, granzyme B, granzyme K, perforin 1 and Fas L were up-regulated from the early stage of vaccination, and peaked at the 3rd w after vaccination with UV-AC. Conclusion： UV-AC of S. japonicum could not ef- fectively induce a Thl response in C57BL/6 mice, which may be an explanation for the low protection against parasite challenge, and the role played by up-regulated expression of cytotoxicity-related genes in mice needs to be further investigated.

Expression on three dsb genes of Chlamydia trachomatis serovar F

To investigate the expression levels of three Dsb protein genes, dsbB, dsbD and dsbG, at different time points post C. trachomatis infection, mouse fibroblast L2 cells were chosen to be infected with C. trachomatis serovar F strain F/IC-Cal-13. C. trachomatis elementary body （EB）-infected L2 cells were harvested immediately after EB attachment onto the cells and every 4 hours post infection （hpi） till 44 hpi for total RNA preparation. RT-PCR assays were then employed to amplify cDNA with primer pairs which are specific to C. trachomatis dsb genes dsbB, dsbD, dsbG and tufA respectively. The relative expression levels of Dsb protein genes were evaluated as cDNA ratios of gene dsb to gene tufA. Our results showed that the transcription of dsbG started from 12 hpi and gradually increased till 44 hpi. The transcription of dsbB and dsbD were detected at 16 hpi and reached their peaks at 28 hpi and 24-28 hpi, respectively. Moreover, there was obvious transcription of dsbB at the later stage （44 hpi）, but none for dsbD at this time point. We came to the conclusion that the expression levels of the three Dsb protein genes are different during the developmental cycle of C. trachomatist. They may play a role in mid-to-late stage of the developmental cycle of C. trachomatis.

DAZ1/DAZ2 cluster deletion mediated by gr / gr recombination per se may not be sufficient for spermatogenesis impairment： a study of Chinese normozoospermic men

Aim： To explore the possible effect of the deleted in azoospermia （DAZ） copy cluster deletion on spermatogenesis in the Chinese population, the deletion of the azoospermia factor c （AZFc） region was analyzed in 346 normozoospermic men. Methods： Three DAZ single nucleotide variant loci and seven AZFc-specific sequence-tagged sites were examined with polymerase chain reaction （PCR）-restriction fragment length polymorphism and routine PCR. Results： Five （1.4%） of the normozoospermic men were found to have deletion of grlgr-DAZ1/DAZ2. None of the men were found to have b2/b4--entire DAZ deletion. Conclusion： The presence of grlgr-DAZ1/DAZ2 deletion in five men with normozoospermia suggests that this deletion per se may not be sufficient for spermatogenic impairment in Chinese men. （Asian J Androl 2006 Mar; 8： 183-187）

Serial changes in expression of functionally clustered genes in progression of liver fibrosis in hepatitis C patients

AIM: To investigate the relationship of changes in expression of marker genes in functional categories or molecular networks comprising one functional category or multiple categories in progression of hepatic fibrosis in hepatitis C (HCV) patients. METHODS: Marker genes were initially identified using DNA microarray data from a rat liver fibrosis model. The expression level of each fibrosis associated marker gene was analyzed using reverse transcription-polymerase chain reaction (RT-PCR) in clinical biopsy specimens from HCV-positive patients (n = 61). Analysis of changes in expression patterns and interactions of marker genes in functional categories was used to assess the biological mechanism of fibrosis. RESULTS: The profile data showed several biological changes associated with progression of hepatic fibrosis. Clustered genes in functional categories showed sequential changes in expression. Several sets of clustered genes, including those related to the extracellular matrix (ECM), inflammation, lipid metabolism, steroid metabolism, and some transcription factors important for hepatic biology showed expression changes in the immediate early phase (F1/F2) of fibrosis. Genes associated with aromatic amino acid (AA) metabolism, sulfur-containing AA metabolism and insulin/ Wnt signaling showed expression changes in the middle phase (F2/F3), and some genes related to glucose metabolism showed altered expression in the late phase of fibrosis (F3/F4). Therefore, molecular networks showing serial changes in gene expression are present in liver fibrosis progression in hepatitis C patients. CONCLUSION: Analysis of gene expression profiles from a perspective of functional categories or molecular networks provides an understanding of disease and suggests new diagnostic methods. Selected marker genes have potential utility for biological identification of advanced fibrosis.

Molecular regulation mechanisms of lymphangiogenesis

Lymphangiogenesis, the growth of new lymphatic vessels, has long been regarded as a putative efficient pathway to neoplastic metastization. Recent results have shown the necessity of lymphatic molecular markers and growth factors for lymphangiogenesis. Importantly, lymphatic endothelial receptor tyrosine kinase VEGFR-3 and its ligands VEGF-C and VEGF-D play crucial roles in promoting lymphatic vascular growth both during development and in pathological conditions. Isolation of pure cultures of lymphatic and blood vascular endothelial cells and systematic characterization of their transcriptomes provide useful cell culture models and novel potential vascular markers and offer further insights into the lymphatic vascular biology. Ectopic expression of the lymphatic endothelial specific homeobox transcription factor Prox1 in blood endothelial cells results in a shift in the gene expression profile towards the lymphatic endothelial phenotype. It demonstrates the plasticity of endothelial cells and offers the possibility of transcriptional reprogramming of vascular endothelial cells as future putative therapeutic applications.

Development of "Multi-Resistance Rice" by Pyramiding of Insect (Cry1C) and Blast Resistance (Pi1 and Pi2) Genes

[Objective] The paper was to improve the blast resistance of insect-resistant transgenic rice. [Method] The Japonica rice variety Jikang10, a new transgenic variety with exogenous insect-resistant gene Cry1C, was used as the receptor, and Kongyu 131, a traditional breeding variety with broad-spectrum high blast resistance genes Pi1 and Pi2, was used as the donor to breeding new rice varieties. The genes were polymerized by hybridization and multi-generation backcrossing, and the offspring of each generation was screened by molecular marker assisted selection, field identification of multi-resistance against insect pests and diseases and agronomic trait selection. [Result] Four lines SK01, SK02, SK03 and SK04 with better resistances to insect pests and blast and outstanding agronomic traits in field were selected. [Conclusion] The results will lay foundations for breeding new multi-resistance rice varieties in Huanghuai rice region.

Computational prediction of over-annotated protein-coding genes in the genome of Agrobacterium tumefaciens strain C58

Agrobacterium tumefaciens strain C58 is a type of pathogen that can cause tumors in some dicotyledonous plants.Ever since the genome of A. tumefaciens strain C58 was sequenced, the quality of annotation of its protein-coding genes has been queried continually, because the annotation varies greatly among different databases. In this paper, the questionable hypothetical genes were re-predicted by integrating the TN curve and Z curve methods. As a result, 30 genes originally annotated as ＂hypothetical＂ were discriminated as being non-coding sequences. By testing the re-prediction program 10 times on data sets composed of the function-known genes, the mean accuracy of 99.99% and mean Matthews correlation coefficient value of 0.9999 were obtained. Further sequence analysis and COG analysis showed that the re-annotation results were very reliable. This work can provide an efficient tool and data resources for future studies of A. tumefaciens strain C58.

Expression Profiles of <i>psbA, ALS, EPSPS</i>, and Other Chloroplastic Genes in Response to PSII-, ALS-, and EPSPS-Inhibitor Treatments in <i>Kochia scoparia</i>

Kochia (Kochia scoparia L. Schrad.), also known as tumbleweed, is an economically important annual C4 broadleaf weed found throughout the US Great Plains. Several herbicides with different modes of action are used in the management of kochia. The effect of commonly used herbicides on the expression of their target site(s) and photosynthetic/chloroplastic genes is poorly understood in weed species, including kochia. The objective of this research was to characterize the expression profiles of herbicide target-site genes, KspsbA, KsALS, and KsEPSPS upon treatment with PSII- (e.g. atrazine), ALS- (e.g. chlorsulfuron), and EPSPS- (e.g. glyphosate)-inhibitors, respectively, in kochia. Furthermore, the expression of genes involved in photosynthesis (e.g. KsRubisco, KsCAB, and KsPPDK) was also determined in response to these herbicide treatments. KspsbA was strongly upregulated (>200-fold) 24 h after atrazine treatment. Transcript levels of the KsALS or KsEPSPS genes were 7 and 3-fold higher 24 h after chlorsulfuron or glyphosate treatment, respectively. KsRubisco, a Calvin cycle gene important for CO2 fixation, was upregulated 7 and 2.6-fold 8 and 24 h after glyphosate and chlorsulfuron treatments, whereas it downregulated 8 and 24 h after atrazine treatment. The transcript levels of KsPPDK remained unchanged after glyphosate treatment but increased 1.8-fold and decreased 2-fold at 24 h after chlorsulfuron and atrazine treatments, respectively. KsCAB remained unchanged after chlorsulfuron treatment, but was downregulated after glyphosate and atrazine treatments. The results show that herbicide treatments not only affect the respective target-site gene expression, but also influence the genes involved in the critical photosynthetic pathway.

Between Scylla and Charybdis:The role of the human immune system in the pathogenesis of hepatitis C

Hepatitis C virus(HCV)frequently elicits only mild immune responses so that it can often establish chronic infection.In this case HCV antigens persist and continue to stimulate the immune system.Antigen persistence then leads to profound changes in the infected host’s immune responsiveness,and eventually contributes to the pathology of chronic hepatitis.This topic highlight summarizes changes associated with chronic hepatitis C concerning innate immunity(interferons,natural killer cells),adaptive immune responses(immunoglobulins,T cells,and mechanisms of immune regulation(regulatory T cells).Our overview clarifies that a strong anti-HCV immune response is frequently associated with acute severe tissue damage.In chronic hepatitis C,however,the effector arms of the immune system either become refractory to activation or take over regulatory functions.Taken together these changes in immunity may lead to persistent liver damage and cirrhosis.Consequently,effector arms of the immune system will not only be considered with respect to antiviral defence but also as pivotal mechanisms of inflammation,necrosis and progression to cirrhosis.Thus,avoiding Scylla-a strong,sustained antiviral immune response with inital tissue damage-takes the infected host to virus-triggered immunopathology,which ultimately leads to cirrhosis and liver cancerthe realm of Charybdis.

Transformation of Two VP1 Genes of O-and Asia 1-Type Foot-and-Mouth Disease Virus into Maize

The expression of antigens in transgenic plants has increasingly been used as an alternative to the classical methodologies for the development of experimental vaccines.This paper reports here the development of a novel oral immunization system for foot-and-mouth disease （FMD） in transgenic maize with two serotypes of the structural protein VP1 of the foot-and-mouth disease virus （FMDV） viz.,O-and Asia 1-type,respectively.The transgenic plantlets were identified and investigated by polymerase chain reaction （PCR）,Southern blot,and real-time PCR.Moreover,it was found that the VP1 genes in transgenic plants could be transmitted stably to the next generation through PCR detection.To our knowledge,this is the first report in an attempt to induce a protective systemic antibody response in animals by feeding the transgenic plants in which two serotypes antigen protein of FMDV expressed together.Results of the experiment provide the possibility of using plant-based vaccines as feedstuff or feedstuff additives.

Selective coding dielectric genes based on proton tailoring to improve microwave absorption of MOFs

Regulating dielectric genes of hollow metal-organic frameworks is a milestone project for microwave absorption(MA).However,there is still a bottleneck in deciphering the contribution of various dielectric genes,making it hard to expand the MA potential from selective encoding gene sequences.Herein,a custom-made proton tailoring strategy is used to build a controllable cavity,and meticulously designed thermodynamic regulation promotes the rearrangement of carbon atoms from disorder to order,thus enhancing the characteristics of charge transfer.Meanwhile,the defect-configuration transformation from heteroatom to vacancy and geometric configuration of hollow structure increase the polarization-related dielectric genes.Therefore,MA performance is enhanced towards broadband absorption(6.6 GHz,1.78 mm)and high-efficiency loss(-62.5 dB),making samples suitable for complex open electromagnetic environments.This work realizes the tradeoff between dielectric gene sequences and provides a profound insight into the functions and sources of various microwave loss mechanisms.

EFFECTS OF SeO_2 ON REGULATORY REGIONS p250 OF c-fos GENE

Objective: This study was designed to determinewhether regulatory regions p250 of c-fos gene wereresponsive to SeO2 and to seek the possible mechanisms of regulation. Methods: HeLa cells were transfected withplasmids p250-tk CAT containing upstream regulatingregions of c-fos gene. Cells were treated by SeO2 for 20 min. CAT expression in transfected cells was observed by thinlayered chromatography. Results: In transfected HeLa cells CAT expression showed obvious increase after exposure to SeO2, especially in 10 mmol/L and 30 mmol/L group (P<0.05). Conclusion: Through affecting regulatory regions p250 of c-fos gene, SeO2 exerted biological effect on tumor cells. SeO2possibly had anti-tumor effects.

蓝萼香茶菜对大鼠全心缺血-再灌注时心肌c-fos基因表达的影响

目的 :研究蓝萼香茶菜对大鼠全心缺血 再灌注时心肌c fos基因表达的影响。方法 :采用Langen dorff离体全心缺血 再灌注损伤模型 ,Wistar大鼠 40只 ,随机分为 5组 ,每组 8只。N组为单纯K H液灌注对照组 ;CN组为缺血 再灌注对照组 ,(高XH ,中XM ,低XL浓度 )蓝萼香茶菜灌注 3组 (浓度分别为 5 % ,1 % ,0 .2 % )。采用免疫组化SP法 ,用相应的c fos抗体检测心肌细胞中c fos基因表达情况。用计算机图像分析系统分析c fos基因表达情况。结果 :与N组相比 ,CN ,XH ,XM ,XL组c fos基因表达水平明显提高 (P <0 .0 1 )。与CN组相比XH ,XM ,XL组c fos基因表达水平明显降低 (P <0 .0 5)。XM ,XL组较XH组c fos基因表达水平明显降低 (P <0 .0 1 )、XM较XL组相比c fos基因表达下降 (P <0 .0 5)。结论 :蓝萼香茶菜能显著抑制心肌c fos基因的表达 ,它保护缺血 再灌注心肌损伤可能部分与其抑制、下调c fos基因的蛋白表达有关。蓝萼香茶菜具有与剂量相关的心肌保护作用 ,但并非浓度越大越有效。

MyoG和c-fos基因多态性及其合并基因型与猪肌纤维性状的关联分析

旨在对MyoG和c-fos基因多态性及其合并基因型与猪肌纤维性状的关联性进行分析。本研究以大白猪(180头)和北京黑猪(170头)2个品种共350头猪作为试验动物,利用冰冻切片、H-E及SDH染色等技术对其进行了肌纤维性状的测定,采用PCR-SSCP技术对肌纤维性状候选基因MyoG与c-fos的外显子进行了SNPs检测,并对不同标记基因型与部分肉质性状和肌纤维性状进行关联分析。结果表明:(1)在MyoG基因外显子1上发现1个多态位点(A2512G),检测到3种基因型(AA、AB、BB);在c-fos基因外显子4上发现2个多态位点(G2650A与A2910G),并分别检测到3种基因型(AA、AB、BB);(2)对A2512G和G2650A2个位点进行分析表明,在2个猪种中,提高A2512G座位上等位基因A的频率可减小肌纤维面积与直径,增加肌纤维密度;提高G2650A位点等位基因B的频率可增加肌纤维密度与红肌纤维比例;而A2910G位点的等位基因A和B则分别具有增加肌纤维面积与肌纤维密度的效应;(3)在北京黑猪,有利合并基因型和不利合并基因型个体的各性状(背膘厚除外)间差异均达到了显著或极显著水平(P<0.05或P<0.01);在大白猪,有利合并基因型和不利合并基因型个体间肌纤维直径、密度、面积、红肌纤维比例、白肌纤维比例和眼肌面积间均达到了显著或极显著水平(P<0.05或P<0.01)。以上结果提示,MyoG和c-fos基因单核苷酸多态性及合并基因型对猪肌纤维性状有一定的影响,可以作为影响猪肌纤维性状的候选基因。