家禽主要组织相容性复合体的研究进展

随着家禽基因组计划的开展 ,对家禽的主要组织相容性复合体 (MHC)的研究取得了较大的进展。关于家禽MHC的各部分基因研究正在逐步深入 ,并且完成了MHC部分测序和染色体定位工作。本文介绍近些年来对家禽MHC的基因结构和作用、与抗体的作用以及相关的基因组研究所取得的进展。人、小鼠及其他动物的相关研究结果将对家禽MHC研究的发展产生重要的影响。

气保焊丝用ER70S-6盘条拉拔断裂原因分析及改进

ER70S-6盘条生产焊丝出现拉拔断裂,通过对断裂样品进行电镜扫描以及金相分析,发现贝氏体和马氏体复合相组织是造成断裂的直接原因。根据该组织形成机制从元素偏析和轧后控冷两个方面分析,提出并实施延长盘条出罩时间、强化盘条保温效果等措施,改善盘条基体组织,解决了断丝问题。

Polymorphism of Exon 3 of MHC Class Ⅱ B Gene in Chinese Alligator(Alligator sinensis)

The polymorphism of MHC class II B gene in 14 Chinese alligators was analyzed, which came from three different areas： a wild population from Xuancheng, Anhui, a captive population from Changxing, Zhejiang, and a captive population from Anhui Research Center for Reproduction of Chinese Alligators. The gene fragment was amplified using a pair of specific primers designed from the MHC gene sequence of the spectacled caiman. A total of 34 sequence haplotypes of exon 3 were detected in the sampled Chinese alligators. The numbers of haplotypes of the 3 Chinese alligator populations were 15, 10, and 9, respectively. The overall estimation of the MHC polymorphism in the Chinese alligator population was higher than those in mammals and in cypdnid fish, The rates of nonsynonymous substitutions （dN） occurred at a significantly lower frequency than that of synonymous substitutions （ds）, which were not consistent with the common rule. This result might suggest that the polymorphism of exon 3 seemed not to be maintained by the balancing selection. The neutrality test of Tajima excluded the null hypothesis that the polymorphism of exon 3 was generated by a random drift, and the fact that D = -0.401 indicated an excess of rare mutations in the Chinese alligator. The nucleotide diversity of the sequences and the phylogenetic relations were also analyzed, and the results suggested that there was no significant difference in genetic diversity among the 3 populations of Chinese alligator.

Microstructure and properties of Al/Cu bimetal in liquid-solid compound casting process

A Ni-P coating was deposited on Cu substrate by electroless plating and the Al/Cu bimetal was produced by solid？liquid compound casting technology. The microstructure, mechanical properties and conductivity of Al/Cu joints with different process parameters （bonding temperature and preheating time） were investigated. The results showed that intermetallics formed at the interface and the thickness and variety increased with the increase of bonding temperature and preheating time. The Ni？P interlayer functioned as a diffusion barrier and protective film which effectively reduced the formation of intermetallics. The shear strength and conductivity of Al/Cu bimetal were reduced by increasing the thickness of intermetallics. In particular, the detrimental effect of Al2Cu phase was more obvious compared with the others. The sample preheated at 780 ℃ for 150 s exhibited the maximum shear strength and conductivity of 49.8 MPa and 5.29×10^5 S/cm, respectively.

新型热成形-淬火碳分配工艺

基于变形、相变、和碳分配同时实现的高强度钢设计思想,利用奥氏体区变形来细化晶粒,利用淬火与分配工艺使钢发生相变且进行碳分配来实现对硬相马氏体和软相残余奥氏体的调控,既一种新型热成形-淬火碳分配碳分配工艺,最终获得纳米级含有错位型马氏体和残余奥氏体的双相复合组织,这种工艺在不损害高强度钢强度的前提下,提高其力学性能,值得推广。

Comparison of human amniotic fluid-derived and umbilical cord Wharton＇s Jelly-derived mesenchymal stromal cells： Characterization and myocardial differentiation capacity

Objective To compare the characterization and myocardial differentiation capacity of arnniotic fluid-derived mesenchymal stromal cells （AF MSCs） and umbilical cord Wharton＇s Jelly-derived mesenchymal stromal cells （WJ MSCs）. Methods The human AF MSCs were cultured from amniotic fluid samples obtained by amniocentesis. The umbilical cord WJ MSCs were obtained from Wharton＇s Jelly of umbilical cords of infants delivered full-term by normal labor. The morphology, growth curves, and analyses by flow cytometry of cell surface markers were compared between the two types of cells. Myocardial genes （GATA-4, c-TnT, a-actin, and Cx43） were detected by real-time PCR and the corresponding protein expressions were detected by Western blot analysis after myocardial induced in AF MSCs and WJ MSCs. Results Our findings revealed AF MSCs and WJ MSCs shared similar morphological characteristics of the fibroblastoid shape. The AF MSCs were easily obtained than the WJ MSCs and had a shorter time to reach adherence of 2.7 ± 1.6 days to WJ MSCs of 6.5 ± 1.8 days. The growth curves by MTT cytotoxic assay showed the AF MSCs had a similar proliferative capacity at passage 5 and passage 10. However, the proliferative capacities ofWJ MSCs were decreased at 5 passage relative to 10 passage. Both AF stem cells and WJ stem cells had the characteristics of mesenchymal stromal cells with some characteristics of embryonic stem cells. They express CD29 and CD105, but not CD34. They were positive for Class I major histocompatibility （MHC I） antigens （HLA-ABC）, and were negative, or mildly positive, for MHC Class II （HLA-DR） antigen. Oct-4 was positive in all the two cells types. Both AF MSCs and WJ MSCs could differentiate along myocardium. The differentiation capacities were detected by the expression of GATA-4, c-TnT, a-actin, Cx43 after myocardial induction. Conclusions Both AF MSCs and WJ MSCs have the potential clinical application for myogenesis in cardiac regenerative therapy.

Microstructure and properties of biodegradable co-continuous(HA+β-TCP)/Zn−3Sn composite fabricated by vacuum casting-infiltration technique

The co-continuous(HA+β-TCP)/Zn−3Sn composite was fabricated via vacuum casting-infiltration method.The microstructure,mechanical properties,corrosion behaviors,and hemolysis ratio of the composite were studied by scanning electron microscope,X-ray diffractometer,mechanical testing,electrochemical test,immersion test,and ultraviolet spectrophotometry.The results indicate that Zn−3Sn alloy infiltrated into porous HA+β-TCP scaffold,which resulted in the formation of a compact(HA+β-TCP)/Zn−3Sn co-continuous composite,without any reaction layer between the Zn−3Sn alloy and the HA+β-TCP scaffold.The compressive strength of the composite was equal to about 3/4 that of Zn−3Sn alloy bulk.The corrosion rate of composite in simulated body fluid solution was slightly higher than that of Zn−3Sn alloy bulk.The main corrosion product on the composite surface was Zn(OH)2.The hemolysis rate of the composite was lower than that of Zn–3Sn alloy bulk and exhibited superior blood compatibility.

Transcriptome of intraperitoneal organs of starry flounder Platichthys stellatus challenged by Edwardsiella ictaluri JCM1680

Platichthys stellatus is an economically important marine bony fish species that is cultured in China on a large scale.However,very little is known about its immune-related genes.In this study,the transcriptome of the immune organs ofP.stellatus that were intraperitoneally challenged with the pathogen Edwardsiella ictaluri JCM1680 is analyzed.Total RNA from four tissues(spleen,kidney,liver,and intestine) was mixed equally and then sequenced on an Illumina HiSeq 2000 platform.Overall,28 465 813 quality reads were generated and assembled into 43 061 unigenes.Similarity searches against public protein sequence databases were used to annotate 28 291 unigenes(65.7%of the total),368 of which were associated with immunoregulation,including 188 related to immunity response.Additionally,the transcript levels of immunity response unigenes annotated as related to tumor necrosis factor(TNF),TNF receptor,chemokine,major histocompatibility complex,and interleukin-6 were investigated in the different tissues of normal and infected P.stellatus by real-time quantitative PCR.The results confirmed that the unigenes identified in the transcriptome database were indeed expressed and up-regulated in infected P.stellatus.To our knowledge,this is the first report of the sequencing and analysis of the transcriptome of P.stellatus.These findings provide insights into the transcriptomics and immunogenetics of bony fish.

Epidemiology and gene markers of ulcerative colitis in the Chinese

Inflammatory bowel disease （IBD） includes two similar yet distinct conditions called ulcerative colitis （UC） and Crohn＇s disease （CD）. These diseases affect the digestive system and cause the inflammation of intestinal tissue, form sores and bleed easily. Most children with IBD are diagnosed in late childhood and adolescence. However, both UC and CD have been reported as early as in infancy. Most information pertaining to the epidemiology of IBD is based upon adult studies. Symptoms include abdominal pain, cramping, fatigue and diarrhea. Genetic factors play a significant role in determining IBD susceptibility. Epidemiological data support a genetic contribution to the pathogenesis of IBD. Recently, numerous new genes have been identified as being involved in the genetic susceptibility to IBD： TNF- 308A, CARD15 （NOD2）, MIF-173, N-acetyltransferase 2 （NAT2）, NKG2D （natural killer cell 2D）, STAT6 （signal transducer and activator of transcription 6）, CTLA-4 （cytotoxic T lymphocyte antigen-4）, MICA-MICB （major histocompatibility complex A and B）, HLA-DRB1, HLA class-Ⅱ, IL-18, IL-4, MICA-A5, CD14, TI R4, Fas-670, p53 and NF-kB. The characterization of these novel genes has the potential to identify therapeutic agents and aid clinical assessment of phenotype and prognosis in patients with IBD （UC and CD）.

Sequence polymorphism of two major histocompatibility(MH)classⅡB genes and their association with Vibrio anguillarum infection in half-smooth tongue sole(Cynoglossus semilaevis)

Major histocompatibility complex （MHC） class II B molecules play an important role in the adaptive immune response in fish. Previous study has reported that two highly polymorphic class II B genes, Cyse-DAB and Cyse-DBB exist in half-smooth tongue sole （Cynoglossus semilaevis）. In this study, the polymorphism within exon 2 of the class II B genes following bacterial challenge was evaluated. Two hundred C. semilaevis individuals were injected intraperitoneally with Vibrio anguillarum. Muscle tissue from the first 20 dead and 20 of the survivors was collected for genotyping. Sixty alleles from the 40 individuals were isolated, of which 32 belonged to Cyse-DAB and 28 belonged to Cyse-DBB. The rate of dN （non-synonymous substitution） was higher than that of ds （synonymous substitution） in the PBRs （peptide binding residues） of both class I1 B genes. Conversely, the rate of ds was higher than dy in the non-PBRs and the complete exon 2 sequence. Thus, the results suggest that positive selection has occurred in the PBRs and purifying selection in the non-PBRs and exon 2. Thirteen class II B alleles were used to study the association between alleles and resistance to infection. Though not significant, alleles Cyse-DAB＊ 0601, Cyse-DAB ＊ 0706, and Cyse-DBB＊O 101, Cyse-DBB＊ 1301 were only found in surviving individuals and may represent alleles that have resistance against V. anguillarum infection. Alleles Cyse-DAB＊0701 and Cyse-DAB＊1301 were significantly more prevalent in dead individuals than in surviving ones and may represent alleles that are associated with increased susceptibility to V. anguillarum infection.

Theoretical basis of a beneficial role for vitamin D in viral hepatitis

Abnormal bone metabolism and dysfunction of the calcium-parathyroid hormone-vitamin D axis have been reported in patients with viral hepatitis. Some studies suggested a relationship between vitamin D and viral hepatitis. Genetic studies have provided an opportunity to identify the proteins that link vitamin D to the pathology of viral hepatitis (i.e., the major histocompatibility complex class Ⅱ molecules, the vitamin D receptor, cytochrome P 450 , the renin-angiotensin system, apolipoprotein E, liver X receptor, toll-like receptor, and the proteins regulated by the Sp1 promoter gene). Vitamin D also exerts its effects on viral hepatitis via non-genomic factors, i.e., matrix metalloproteinase, endothelial vascular growth factor, prostaglandins, cyclooxygenase-2, and oxidative stress. In conclusion, vitamin D could have a beneficial role in viral hepatitis. Calcitriol is best used for viral hepatitis because it is the active form of the vitamin D 3 metabolite.

ENDOGENOUS EXPRESSION AND HLA STABILIZATION ASSAY OF PLASMODIUM FALCIPARUM CTL EPITOPE MINIGENE IN HUMAN HLA- A2.1 AND HLA- B51 CELLS

To evaluate the Plasmodium falciparum CTL epitope vaccines in HLA class I allele specific human cell lines that have high frequency among Chinese population. Methods. Synthesized oligonucleotides encoding for P.f. CTL epitope genes, constructed eukaryotic expression plasmids, transfected the minigenes into HLA class I allele specific human cell lines and identified endogenous expressing of the minigenes by RT- PCR and HLA stabilization assay. Results. Two mini- genes encoding Plasmodium falciparum CTL epitopes were designed and cloned, respectively, into an eukaryotic expressing vector to form TR26 which was restricted to HLA- B51, SH6 which was restricted to HLA- A2.1, and TS, which had the two aforementioned mini- genes fused in tandem. All of these CTL epitope genes were transfected and endogenously expressed in respective cell lines containing appropriate HLA molecules. The obviously increased expressions of HLA class I molecules were detected in the transfected cell lines. It was demonstrated that the two discrete Plasmodium falciparum epitope genes were effectively processed and presented, and the close proximity of the two epitope genes in one chain as in mini- gene TS did not interfere with the processing and presenting of each epitope gene in corresponding cell line. Conclusion. A successful expression and presentation of multiple CTL epitope mini- gene in MHC class I allele specific human cell lines were demonstrated by an in vitro assay, which could be corresponding to the vaccination of CTL vaccines in people with different MHC I molecules. This work also suggested the possibility of constructing a multiple CTL epitope plasmodium falciparum DNA vaccine that could cover most of Chinese population.

Variation of DRB1 Gene in Tibetan Sheep

It reveals that the MHC （major histocompatibility complex） gene product always involved in the control of immune response and disease resistance. Nowadays many studies have indicated the OLA （ovine lymphocyte antigen） DRB1 gene is associated with some sheep diseases. Tibetan sheep is one of the three major shag sheep breeds in China, and also have the largest number of China＇s sheep breeds. But till now no report has been seen on studying DRB1 gene in Tibetan sheep of China. To understand the evolution and provide the basis for sheep disease resistance, polymorphism in the exon2 ofDRB1 gene in Tibetan sheep was analyzed. The PCR-SSCP, cloning and sequencing were used to analyse DRB1 gene variation in 600 Tibetan sheep of China. And the genetic relationship and evolutionary significance of the alleles had also been analyzed. Total of 31 alleles were identified, in which 15 alleles had not been reported before. And there were 70 SNPs （single nucleotide polymorphisms） sites in 31 sheep DRB1 gene haplotypes, the proportion was 29.5% to the whole exort2 sequence. All of this indicated that DRB1 exon2 is highly polymorphic in Tibetan sheep. The variation identified here might have an impact on both the function and level of expression of the OLA-DRB1.

Identification of two Major Histocompatibility(MH) Class Ⅱ A Genes and Their Association to Vibrio anguillarum Infection in Half-smooth Tongue Sole(Cynoglossus semilaevis)

Major histocompatibility complex class II antigens are important in vertebrate immune system.In the present study,the full cDNA sequence of class II A gene was synthesized by RACE-PCR from half-smooth tongue sole(Cynoglossus semilaevis),and its open reading frame(ORF) polymorphism was studied.The whole cDNA sequence was 992 bp in length,including the ORF with 717 bp.Twenty-five alleles were identified and clustered into two distinct groups according to the specific nucleotides/amino acids in specific positions.Eleven alleles belonged to Cyse-DAA while fourteen alleles belonged to Cyse-DBA.Four Cyse-DAA alleles were observed in one individual,and three to five Cyse-DBA alleles were observed in each of the three detected individuals,which indicated that at least two loci existed in each gene.Moreover,in order to study the function of the alleles in resistance to infection,200 individuals were intraperitoneally injected with Vibrio anguillarum and the first 20 dead individuals and 20 surviving ones were selected for genotype analysis.Fifty-six alleles were identified among the 40 individuals.Twenty-nine alleles belonged to Cyse-DAA and the other 27 alleles belonged to Cyse-DBA.Eighteen alleles were selected for studying their function in resistance to infection.Alleles Cyse-DAA*0201,Cyse-DAA*1101,Cyse-DBA*0401,Cyse-DBA*1102,Cyse-DBA*1801 and Cyse-DBA*2201 were identi-fied only in surviving individuals,while alleles Cyse-DAA*0901,Cyse-DBA*1101 and Cyse-DBA*1401 occurred more frequently in dead individuals.This study confirmed the existence and polymorphism of two class II A genes as well as the relationship between alleles of class II A genes and disease susceptibility/resistance in half-smooth tongue sole.

Relationship between HLA-DRB1 and DQ alleles and the genetic susceptibility to type 1 diabetes

Objective To study the relationship between human leukocyte antigen (HLA) DRB1 and DQ alleles and the genetic susceptibility of type 1 diabetes in North Chinese children Methods Polymerase chain reaction (PCR) techniques were used to amplify the second exon of DRB1 and DQ alleles, after which sequence specific olignucleotide probe (SSOP) dot blot hybridization techniques were used to analyze the amplified products Results DRB1*0301, DQA1*0301, DQB1*0201 alleles and DRB1*0301 DQA1*0501 DQB1*0201 haplotype were significantly increased in patients, while DQA1*0103 and DQB1*0601 alleles were significantly increased in controls The distribution of DR4 and DR9 haplotypes in patients and controls were not significantly different, but DR3/DR4 and DR4/DR9 heterozygotes were significantly increased in patients Conclusions DRB1*0301, DQA1*0301 and DQB1*0201 confer susceptibility while DQA1*0103 and DQB1*0601 confer protection to type 1 diabetes DRB1*0301 DQA1*0501 DQB1*0201 haplotype offers a predisposition to type 1 diabetes in North Chinese Although the distribution of DR4 and DR9 in patients and controls had no significant difference, DR3/DR4 and DR3/DR9 heterozygotes were significantly increased in patients, showing that the susceptive effects of DR3 and DR4 or DR4 and DR9 haplotypes could be added up

NK cell-based cancer immunotherapy: from basic biology to clinical application

Natural killer(NK) cells, which recognize and kill target cells independent of antigen specificity and major histocompatibility complex(MHC) matching, play pivotal roles in immune defence against tumors. However, tumor cells often acquire the ability to escape NK cell-mediated immune surveillance. Thus, understanding mechanisms underlying regulation of NK cell phenotype and function within the tumor environment is instrumental for designing new approaches to improve the current cell-based immunotherapy. In this review, we elaborate the main biological features and molecular mechanisms of NK cells that pertain to regulation of NK cell-mediated anti-tumor activity. We further overview current clinical approaches regarding NK cell-based cancer therapy, including cytokine infusion, adoptive transfer of autologous or allogeneic NK cells, applications of chimeric antigen receptor(CAR)-expressing NK cells and adoptive transfer of memory-like NK cells. With these promising clinical outcomes and fuller understanding the basic questions raised in this review, we foresee that NK cell-based approaches may hold great potential for future cancer immunotherapy.

Microstructure and mechanical properties of liquid phase sintered silicon carbide composites

Silicon carbide (SiC) composites were prepared by hot-press sintering from α-SiC starting powders with BaAl2Si2O8 (BAS). The effects of additives on densification, microstructure, flexural strength, and fracture behavior of the liquid phase sintered (LPS) SiC composites were investigated. The results show that the served BAS effectively promotes the densification of SiC composites. The flexural strength and fracture toughness of the SiC composites can reach a maximum value of 454 MPa and 5.1 MPa·m1/2, respectively, for 40% (w/w) BAS/SiC composites. SiC grain pullout, crack deflection, and crack bridging were main toughening mechanisms for the sintered composites.

MHC polymorphism and disease resistance to Singapore grouper iridovirus(SGIV) in the orange-spotted grouper, Epinephelus coioides

Major histocompatibility complex （MHC） genes are critical members in both innate and adaptive immunity, and the association between their polymorphism and disease resistance has been reported in many teleosts. In the present study, we first investigated the genetic variation at the MHC II β gene in orange-spotted grouper （Epinephelus coioides） after a challenge with Singapore grouper iridovirus （SGIV）. The results reveal that a high polymorphism level of the MHC II β gene （H = 1.000; K = 20.206; π=0.081） and at least three loci exist in grouper. The rate of dN/dS in the peptide-binding region （PBR） and non-PBR were both 〉1, suggesting the loci were evolving under positive selection. A high ratio of heterozygous individuals （37.26 %） and high rate of dN/dS were discovered, suggesting that both heterozygote advantage and frequency-dependent selection might result in the high polymorphism levels in MHC II β genes in grouper. A total of 33 MHC II β alleles were identified from 40 high-susceptibility （HS） and 40 high-re- sistance group （HR） individuals, and 15 alleles were used in the association analysis. Three alleles, EPCO-DBB＊0302, EPCO-DBB＊0307, EPCO-DBB＊0603, and EPCO- DBB＊1001 were significantly associated with resistance ability to SG1V, and the EPCO-DBB＊0607 and EPCO-DBB＊1303 alleles were associated with susceptibility （P 〈 0.05）. To further confirm the association, another independent challenge experiment was performed. The result of association analysis in the verification test found that only EPCO-DBB＊1001 alleles were significantly asso- ciated with resistance to SGIV （P 〈 0.05）, while the other alleles showed no significance （P 〉 0.05） in the frequency distribution between HR and HS groups. Therefore, the EPCO-DBB＊ 1001 alleles could be used as a disease resis- tance-related MHC marker in the molecular marker-assisted selective breeding program of grouper.

Balancing selection and recombination drive genetic variation at MHC class I genes in the giant panda

Major histocompatibility complex （MHC） is a family of highly polymorphic genes activating adaptive immunity in vertebrates. However, the underlying mecha- nism of MHC evolution is still not fully understood. In this study, we investigated genetic variation of three classical MHC class I genes in the giant panda （Ailuropoda mela- noleuca） and tested for selection effect and recombination event across exonic and intronic sequences to understand maintenance mechanism of polymorphism at Aime-MHC class I genes. In total, we isolated 21 MHC class I haplotypes （exon 2-intron 2-exon 3） from 46 captive giant pandas, of which eight were for Aime-C, seven for Aime-I and six for Aime-L; however, we only identified six unique sequences from these haplotypes. The subsequent maximum-likeli- hood and Chi-square analyses both detected evidence of recombination acting on the 21 haplotypes. These results indicate that the giant panda still retains a relatively high adaptive variation at Aime-MHC-I genes, and that the intronic segments have been homogenized along evolu- tionary time by recombination and subsequent genetic drift.We calculated nucleotide substitution rates of the antigen- binding regions （exons 2 and 3） and the noncoding intron 2, and found two pieces of evidence supporting the presence of balancing selection in the giant panda： an excess of nonsynonymous over synonymous substitutions at the antigen-binding sites, and an obviously higher synonymous substitutions in the exons than nucleotide substitutions in the intron. Thus, this study reveals that balancing selection and recombination together shape the diversity pattern at Aime- MHC-I loci of the giant panda.