Evaluation on Spot Weld Models in Structural Dynamic Analysis of Automotive Body in White

Spot weld models are widely used in finite element analysis（FEA） of automotive body in white（BIW） to predict static,dynamic,durability and other characteristics of automotive BIW.However,few researches are done on evaluation of the validity of these spot weld models in structural dynamic analysis of BIW.To evaluate the validity and accuracy of spot weld models in structural dynamic analysis of BIW,two object functions,error function and deviation function,are introduced innovatively.Modal analysis of Two-panel and Double-hat structures,which are the dominated structures in BIW,is conducted,and the values of these two object functions are obtained.Based on the values of object functions,the validity of these spot weld models are evaluated.It is found that the area contact method（ACM2） and weld element connection（CWELD） can give more precise prediction in modal analysis of these two classical structures,thus are more applicable to structural dynamic analysis of automotive BIW.Modal analysis of a classical BIW is performed,which further confirms this evaluation.The error function and deviation function proposed in this research can give guidance on the adaptability of spot weld models in structural dynamic analysis of BIW.And this evaluation method can also be adopted in evaluation of other finite element models in static,dynamic and other kinds of analysis for automotive structures.

SEM Comparison of Penetration in Artificial White Spots Lesion between an Infiltrant Resin and Two Adhesive Systems

White spot infiltration emerged as an alternative of non-invasive treatment to halt progression of the lesion, through the use of low viscosity resins that would permeate the porous enamel and form a physical barrier that would prevent the acid diffusion produced by micro-organisms. Purpose: To compare penetration levels in artificial white spot lesions, of infiltrant resin ICON™and 2 conventional adhesives systems, XP-Bond™and Single Bond 2™. Methodology: White spot lesions (ICDAS code 2) were caused in 75 premolars or third molars were extracted in good conditions, by immersion in a 0.1 M lactic acid solution (pH 4.5) at 37℃ for 8 weeks. They were divided randomly into 3 groups of 25 samples and applied the following resins, Group A: ICON™, B: XP-Bond™and C: Single Bond 2™. Subsequently, the enamel was removed with hydrochloric acid to expose resin saturated area and the samples were metalized with Au-Pd for SEM observation. The resin tags lengths were measured on microphotographs through software, and the values were analyzed with the statistics ANOVA and Scheffé post-test. Results: There were significant differences (p ™(82.7 μm ± 26.8 μm) compared to adhesive systems XP-Bond™(58.5 μm ± 29.3 μm) and Single Bond 2™(44.8 μm ± 32.5 μm). We found no significant differences between the two adhesive systems (p > 0.05). Conclusion: Under the conditions tested, the penetration of infiltrant ICON was significantly higher than the adhesive systems;however, it removes the surface layer of the enamel.

Development and application of antibody microarray for white spot syndrome virus detection in shrimp

Detecting white spot syndrome virus （WSSV） in shrimp in high efficiency and veracity is important for disease prevention in aquaculture. Antibody-based mieroarray is a novel proteomic technology that can meet the requirements. In this study, we developed an antibody microarray for WSSV-detection in a specific and parallel way at multiple samples. First, seven slides each with different modifications were characterized by atomic force microscope, and were compared in the efficiency of immobilizing proteins. Of the seven, 3-dimensional structured agarose gel-modified slides were chosen appropriate for the microarray for having higher signal value and superior spot size. A purified rabbit anti-WSSV antibody was arrayed as the capture antibody of the microarray on the agarose gel-modified slides, and then the mieroarray slides were incubated in the tissue homogenate of sampled shrimp and the antibody-antigen complex was detected by Cy3-conjugated anti-WSSV monoclonal antibody. The results were measured by a laser chipscanner and analyzed with software. To obtain satisfied fluorescence signal intensity, optimal conditions were searched. The detection limit of the antibody microarray for WSSV is 0.62μg/mL, with a woven long shelf life for 6 months at 4℃ or 8 months at -20℃. Furthermore, concordance between antibody microarray and traditional indirect ELISA reached 100% for WSSV detection. These results suggest that the antibody microarray could be served as an effective tool for diagnostic and epidemiological studies of WSSV.

Proteomic Analyses of the Shrimp White Spot Syndrome Virus

White spot syndrome virus (WSSV), a unique member within the virus family Nimaviridae, is the most notorious aquatic virus infecting shrimp and other crustaceans and has caused enormous economic losses in the shrimp farming industry worldwide. Therefore, a comprehensive understanding of WSSV morphogenesis, structural proteins, and replication is essential for developing prevention measures of this serious parasite. The viral genome is approximately 300kb and contains more than 180 open reading frames (ORF). However, most of proteins encoded by these ORF have not been characterized. Due to the importance of WSSV structural proteins in the composition of the virion structure, infection process and interaction with host cells, knowledge of structural proteins is essential to understanding WSSV entry and infection as well as for exploring effective prevention measures. This review article summarizes mainly current investigations on WSSV structural proteins including the relative quantities, localization, function and protein-protein interactions. Traditional proteomic studies of 1D or 2D gel electrophoresis separations and mass spectrometry (MS) followed by database searches have identified a total of 39 structural proteins. Shotgun proteomics and iTRAQ were initiated to identify more structural proteins. To date, it is estimated that WSSV is assembled by at least 59 structural proteins, among them 35 are defined as the envelope fraction (including tegument proteins) and 9 as nucleocapsid proteins. Furthermore, the interaction within several major structural proteins has also been investigated. This identitification and characterization of WSSV protein components should help in the understanding of the viral assembly process and elucidate the roles of several major structural proteins.

Impact of Vibrio parahaemolyticus and white spot syndrome virus(WSSV)co-infection on survival of penaeid shrimp Litopenaeus vannamei

White spot syndrome virus(WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect L itopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus(WSSV+Vp), or we fi rst infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection(Vp+WSSV). The effect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of V ibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction(PCR) method. L v My D88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the different mortality rates. Our results show that(1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection;(2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed signifi cant increases in the numbers of Vibrio( P <0.05);(3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 10 5 to 10 7 /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V. parahaemolyticus and WSSV led to massive death of L. vannamei.

White spot syndrome virus envelope protein VP124 involved in the virus infection

White spot syndrome virus （WSSV） is one of the major shrimp pathogens causing large economic losses to shrimp farming. In an attempt to identify the envelope proteins involved in the virus infection, purified WSSV virions were mixed with three antisera against WSSV envelope proteins （VP39, VP124 and VP187 ）, individually. And then they were injected intramuscularly into crayfish （Procambarus clarkii） to conduct in vivo neutralization assays. The results showed that for groups injected with virions only and groups injected with the mixture of virions and antiserum against VP124, the crayfish mortalities were 100% and 60% on the 8th day postinfection, individually. The virus infection could be delayed or neutralized by antibody against the envelope protein VP124. Quantitative PCR was used to further investigate the influence of three antisera described above on the virus infection. The results showed that the antiserum against VP124 could restrain the propagation of WSSV in crayfish. All of the results suggested that the viral envelope protein VP124 played a role in WSSV infection.

Construction of white spot syndrome virus (WSSV) whole genome phage display library

A rebuilt vector pCANTAB 5 EE was obtained by inserting a 34 bp double-stranded oligonucleotide which contained a EcoRV recognition sequence into pCANTAB 5 E. White spot syndrome virus （WSSV） genome DNA was fragmented by sonication to isolate fragments mainly in the range of 0.8 ～2.0 kb, then the fragments were blunt-ended with T4 DNA polymerase and cloned into the EcoRV site of pCANTAB 5 EE. The primary recombinant clone of the library was 3.0 × 10^5.Colony PCR of random selected recombinants showed that the size of the inserts was 0.12 ～ 1.77 kb. After the whole library recombinant phages infected Escherichia coli HB2151 cells, the extracellular and periplasmic extracts were dropped on PVDF membranes to perform dot blot, using polyclonal mouse anti-VP24 serum,anti-WSV026 serum,anti-WSV063 serum,anti-WSV069 serum,anti-WSV112 serum, anti WSV238 serum,anti-WSV303 serum and anti-VP26 serum as the primary antibody, respectively. The results showed that the display library could express the viral proteins.

White spot syndrome virus inactivation study by using gamma irradiation

The present study was conducted to investigate the effect of gamma irradiation on white spot syndrome virus （WS SV）. White spot syndrome virus is a pathogen of major economic importance in cultured penaeid shrimp industries. White spot disease can cause mortalities reaching 100% within 3-10 days of gross signs appearing. During the period of culture, immunostimulant agents and vaccines may provide potential methods to protect shrimps from opportunistic and pathogenic microrganisms. In this study, firstly, WSSV was isolated from infected shrimp and then multiplied in crayfish. WSSV was purified from the infected crayfish haemolymph by sucrose gradient and confirmed by transmission electron microscopy. In vivo virus titration was performed in shrimp, Penaeus semisulcatus. The LD50 of live virus stock was calculated 1054/mL. Shrimp post-larvae （1-2 g） were treated with gamma-irradiated （different doses） WSSV （10^o to 10^-4 dilutions） for a period of 10 days. The dose/survival curve for irradiated and un-irradiated WSSV was drawn; the optimum dose range for inactivation of WSSV and unaltered antigenicity was obtained 14- 15 kGy. This preliminary information suggests that shrimp appear to benefit from treatment with gamma- irradiated WSSV especially at 14-15 KGy.

Minicistron from a gene of prawn white spot bacilliform virus(WSBV)and its expression

By the sequencing and analysis of genomic DNA and cDNA of WSBV, it was found that there existed a minicistron in the leading sequence of p204 gene. The minicistron utilized rare codons of genes from prawn and WSBV. The p204 gene could be expressed in Escherichia coli whether a miniciston exited in its leading sequence or not, but the amounts of the expressed products of them were different. The minicistron might be related to the expression of p204 gene.

Production and Characterization of Monoclonal Antibodies of Shrimp White Spot Syndrome Virus Envelope Protein VP28

BALB/c 老鼠与净化的白点症候群病毒(WSSV ) 被使免疫。六根 monoclonal 抗体房间线被 ELISA 选择， VP28 蛋白质在 E 表示了。coli。在 vitro 中立化，实验证明他们中的 4 个能在喇蛄禁止病毒感染。西方污点建议所有这些 monoclonal 抗体对 VP28 的 conformational 结构。monoclonal 抗体 7B4 用胶体的金粒子被标记并且过去常由标记的 immunogold 在病毒信封上定位 VP28。这些 monoclonal 抗体能被用来为 WSSV 感染开发免疫学的诊断方法。关键词怀特点症候群病毒(WSSV )- Mab - 信封 - 本地化 - 中立化 CLC 数字 S945.

Peritrophin-like protein from Litopenaeus vannamei (LvPT) involved in white spot syndrome virus (WSSV) infection in digestive tract challenged with reverse gavage

The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus （WSSV） in shrimp. In this study, the nucleotide sequence encoding peritrophin-like protein of Litopenaeus vannamei （LvPT） was amplified from a yeast two-hybrid library of L. vannamei. The epitope peptide of LvPT was predicted with the GenScript OptimumAntigenTM design tool. An anti-LvPT polyclonal antibody was produced and shown to specifically bind a band at -27 kDa, identified as LvPT. The LvPT protein was expressed and its concentration determined. LvPT dsRNA （4 pg per shrimp） was used to inhibit LvPT expression in shrimp, and a WSSV challenge experiment was then performed with reverse gavage. The pleopods, stomachs, and guts were collected from the shrimp at 0, 24, 48, and 72 h post-infection （hpi）. Viral load quantification showed that the levels of WSSV were significantly lower in the pleopods, stomachs, and guts of shrimp after LvPT dsRNA interference than in those of the controls at 48 and 72 hpi. Our results imply that LvPT plays an important role during WSSV infection of the digestive tract.

Characterization and Diagnostic Use of a Monoclonal Antibody for VP28 Envelope Protein of White Spot Syndrome Virus

The gene encoding the VP28 envelope protein of White spot syndrome virus (WSSV) was cloned into expression vector pET-30a and transformed into the Escherichia coli strain BL21.After induction,the recombinant VP28 (rVP28) protein was purified and then used to immunize Balb/c mice for monoclonal antibody (MAb) production.It was observed by immuno-electron microscopy the MAbs specific to rVP28 could recognize native VP28 target epitopes of WSSV and dot-blot analysis was used to detect natural WSSV infection.Competitive PCR showed that the viral level was approximately 104 copies/mg tissue in the dilution of gill homogenate of WSSV-infected crayfish at the detection limit of dot-blot assay.Our results suggest that dot-blot analysis with anti-rVP28 MAb could rapidly and sensitively detect WSSV at the early stages of WSSV infection.

Identification and Characterization of Nuclear Localization Signals within the Nucleocapsid Protein VP15 of White Spot Syndrome Virus

The nucleocapsid protein VP15 of white spot syndrome virus (WSSV) is a basic DNA-binding protein. Three canonical bipartite nuclear localization signals (NLSs), called NLS1 (aa 11-27), NLS2 (aa 33-49) and NLS3 (44-60), have been detected in this protein, using the ScanProsite computer program. To determine the nuclear localization sequence of VP15, the full-length open reading frame, or the sequence of one of the three NLSs, was fused to the green fluorescent protein (GFP) gene, and transiently expressed in insect Sf9 cells. Transfection with full-length VP15 resulted in GFP fluorescence being distributed exclusively in the nucleus. NLS1 alone could also direct GFP to the nucleus, but less efficiently. Neither of the other two NLSs (NLS2 and 3) was functional when expressed alone, but exhibited similar activity to NLS1 when they were expressed as a fusion peptide. Furthermore, a mutated VP15, in which the two basic amino acids (11RR12) of NLS1 were changed to two alanines (11AA12), caused GFP to be localized only in the cytoplasm of Sf9 cells. These results demonstrated that VP15, as a nuclear localization protein, needs cooperation between its three NLSs, and that the two residues (11RR12) of NLS1 play a key role in transporting the protein to the nucleus.

Interaction between a Peptide and White Spot Syndrome Virus VP28 Envelope Protein

White spot syndrome virus (WSSV) is one of the most important pathogens that endanger the global shrimp aquaculture. Studies have confirmed that in the early stage of infection, VP28, the main envelope protein of WSSV, is used as a viral adhesion protein to bind PcRab7 of Penaeus chinensis, helping virus enter the host cells, resulting in shrimp infection. Hence, inhibition of envelope protein VP28 would be a novel way to deal with the infection. Peptide 2E6 was confirmed to have a high specificity and blocked virus infection. However, the mechanism by which it combines with VP28 is not clear. Clarifying the binding mechanism between peptides and VP28 is of great significance for further optimization and screening of antiviral peptides. In this research, the MD simulation and binding free energy analysis were implemented to validate and capture intermolecular interactions aims to clarify the blocking mechanism.

Homozygous lethality and heterozygous spotting due to a novel missense mutation in the mouse Kit gene

N-ethyl-N-nitrosourea （ENU） mutagenesis in mice can be used to study gene function in vivo and to establish genetic mouse models of human disease. In this study, a white spotted mouse （named Kit^W-1 Bao） was obtained by ENU-induced mutagenesis. Inheritance testing showed a single-gene dominant mutation and lethality in the Kit^W-1 Bao homozygous mice. The mutation was mapped to Chromosome 5 between markers DSMit356 and DSMit308. The region contains the Kit gene, whose mutations are known to lead to pigmentation defects in mice. Sequence analysis of the Kit cDNA from Kit^W-1 Bao heterozygotes revealed an A to T missense mutation resulting in an amino acid substitution of Asp （D） by Val （V） at amino acid position 849 within a highly conserved tyrosine kinase domain. The combined phenotype displayed by the Kit^W-1 Bao heterozygous and homozygous mutant mice demonstrates the critical function of the highly conserved aspartie acid residue at position 849 in the Kit gene product

玉米白斑病的症状识别与诊断

由宽颈附球孢菌Epicoccum latusicollum Qian Chen, Crous&L. Cai引起的玉米白斑病正在我国局部地区流行,导致重大损失,并有扩大流行范围的趋势。为了增加植保及玉米抗病育种工作者对玉米白斑病的了解,本文描述了该病害的症状特点、病原菌特征,总结了发生规律、病害循环及分离鉴定和诊断技术。玉米白斑病菌能在病残体上越冬,种子带菌是其远距离传播的主要途径,风夹雨、昆虫及农事操作人员活动是近距离传播途径。7月中下旬至9月中下旬是白斑病流行高峰期,病菌与玉米适宜生长温度基本一致,具有扩散传播的风险。

凡纳对虾核心育种群生长和抗WSSV性状的遗传参数估计

为开展凡纳对虾(Penaeus vannamei)生长和白斑综合征病毒(white spot syndrome virus,WSSV)抗性复合选育,本研究以凡纳对虾59个核心育种群家系1770尾77~94日龄的个体为实验材料,利用两性状动物模型、公母畜阈值模型,评估在WSSV感染情况下凡纳对虾体长、抗WSSV存活时间和家系WSSV半致死存活率的遗传力和遗传相关。结果显示,凡纳对虾体长、抗WSSV存活时间和家系WSSV半致死存活率遗传力估计值分别为0.17±0.05、0.18±0.05和0.14±0.05,均属于中等遗传力水平,且均与0差异极显著(P<0.01)。凡纳对虾体长与抗WSSV存活时间性状的遗传相关系数为0.15±0.20,与家系WSSV半致死存活率性状的遗传相关系数为0.25±0.22,上述遗传相关与0差异不显著(P>0.05);抗WSSV存活时间性状与家系WSSV半致死存活率性状的遗传相关系数为0.96±0.03,遗传相关与1差异不显著(P>0.05),为高度正相关。结果表明,在该育种群体中,凡纳对虾生长与WSSV抗性可根据育种需要,通过赋值制定综合选择指数,进行多性状复合选育。此外,为优化每代育种的操作过程,可选用家系WSSV半致死存活率作为WSSV抗性的指标性状。本研究为开展凡纳对虾生长和WSSV抗性优良品种的选育提供了基础数据和理论支撑。

隐形与固定矫治器正畸后白垩斑发病率的比较

目的:通过回顾性研究比较使用固定矫治器和隐形矫治器正畸后白垩斑发病率。方法:收集华西口腔医院正畸科完成正畸治疗的患者资料,按照矫治器类型的不同分为隐形矫治器组与固定矫治器组。同一患者治疗前后的口内照片被并排放置在电脑显示器上比较,治疗前不存在,治疗后出现的釉质不透明白色斑块被确定为正畸后白垩斑。使用SPSS软件进行数据统计,分析两组患者白垩斑发病率的差异。结果:隐形矫治器组白垩斑的发病率为4.63%,显著低于固定矫治器组(17.27%,P=0.003)。此外,在所有相关因素中,男性患者比女性患者更容易发生白垩斑(P=0.021);青少年的发病率显著高于成人(P<0.001),同时治疗时间超过25个月的患者更有可能出现为白垩斑(P<0.001)。治疗后口腔卫生较好的患者白垩斑发病率低(P<0.001)。结论:使用隐形矫治器的患者正畸后白垩斑的发病率低于使用固定矫治器的患者。同时,青少年、男性、口腔卫生差、治疗时间长是正畸后白垩斑病变的危险因素。

不同杀菌剂对玉米白斑病的田间防治效果

玉米白斑病是我国西南玉米产区一种新发病害,尚缺乏有效的防控措施。为筛选玉米白斑病的防控药剂,本研究选用19种不同作用机制的杀菌剂,于2022年-2023年在云南省普洱市开展田间试验,分别于玉米10叶期和抽雄吐丝期采用叶面喷雾法施药1次,评价各药剂的田间防效。结果显示,9种防治真菌性病害的杀菌剂在最后1次施药后28 d对白斑病的防治效果超过50%,包括250 g/L嘧菌酯悬浮剂、40%氟环唑悬浮剂、24%井冈霉素A水剂、40%咪鲜胺水乳剂和10%苯醚甲环唑微乳剂5种单剂,以及325 g/L苯甲·嘧菌酯悬浮剂、40%丁香·戊唑醇悬浮剂、40%唑醚·戊唑醇悬浮剂和30%氟环·咪鲜胺微乳剂4种混剂。保护性杀菌剂代森锰锌和有机铜杀菌剂喹啉铜仅表现了23.26%和29.00%的微弱防效。卵菌病害特效药剂烯酰吗啉,防治细菌性病害的杀菌剂春雷霉素、四霉素、中生菌素以及植物免疫诱抗剂6%寡糖·链蛋白可湿性粉剂的防效较差,均低于20%。本研究发现部分甲氧基丙烯酸酯类和三唑类杀菌剂对玉米白斑病具有较好的田间防治效果,建议现阶段在生产上采用二者的复配药剂如苯甲·嘧菌酯,唑醚·戊唑醇等开展田间防控。

青少年正畸固定矫治中釉质健康与脱矿状态下龈上菌斑真菌微生物组的比较研究

目的牙釉质脱矿是固定矫治常见的并发症。本研究比较了固定矫治中釉质脱矿与健康状态下龈上菌斑真菌组的差异,探索口腔真菌与正畸釉质脱矿的关系。方法选取54名固定矫治患者,釉质脱矿组31人,健康对照组23人。收集龈上菌斑,通过转录间隔子测序比较两组真菌组差异,使用RT-qPCR分析白色念珠菌的丰度变化。结果两组真菌微生物多样性和优势物种丰度均存在显著差异。脱矿组真菌微生物多样性降低,念珠菌属及白色念珠菌在脱矿组中富集,且白色念珠菌在脱矿组中丰度较高(P<0.05)。结论青少年固定矫治脱矿状态下龈上菌斑真菌组与健康状态存在显著差异,白色念珠菌更易富集在脱矿菌斑中,可能与釉质脱矿的发生发展密切相关。