The Structure and Function of the Bunyavirus Nucleoprotein / 中国生物化学与分子生物学报

Bunyavirus is widely distributed, highly contagious, and has a high fatality rate. It is a negative- strand RNA virus that has a major impact on public health around the world. The development of vaccines and the search for drugs are the key to prevent bunyavirus infection. The nucleoprotein (NP) of viruses is necessary for the synthesis of viral RNA, which combines with viral RNA to form the nucleocapsid, participates in viral assembly and RNA transcription, and plays an important role in viral proliferation. In addition, NP also has B cell and T cell epitopes, which can induce cellular and humoral immunity, so NP is an ideal target for vaccine design and drug development. Given its abundance and specificity, NP is also commonly used in the detection of viral diseases. More and more bunyavirus NP structures and structures of NP-RNA complexes have been resolved. Researchers have discovered two important antiviral targets through these structures, the terminal arm and the RNA binding cleft. This paper reviews the function and three-dimensional structure of the bunyavirus NP and the research progress of NP as an antiviral target, in order to provide a theoretical basis for the prevention and treatment of the bunyavirus disease.

Enantiomeric characteristics of non-synthetic cannabidiol by two-dimensional nuclear magnetic resonance

SUMMARY Introduction: Cannabidiol (CBD) has become a promising bioactive for the next decades after the recent recognition of the medical potential of Cannabis derivatives by United Nations member countries, as it has no psychotropic potential as your isomer A9-tetrahydrocannabinol (Δ9-THC). The differentiation of these isomers has been studied for decades. Recent studies demonstrate that even with more subtle chemical characteristics, such as those of the CBD enantiomers, there are considerable bioactive differences. However, there are still not many studies on their chemical structures. Aim: This work aims to present experimental data obtained by Nuclear Magnetic Resonance (NMR) to better elucidate the three-dimensional structure of this enantiomeric bioactive. Materials and methods: For this, a sample of non-synthetic high purity CBD was subjected to different one-dimensional (1D-NMR) and two-dimensional (2D-NMR) analyses related to the hydrogen (1H) and carbon (13C) nuclei. Results and discussion: The 1D-NMR techniques used are sufficient to distinguish the CBD and Δ 9-THC isomers, but not to identify the enantiomeric characteristics of the non-synthetic CBD. Conclusions: It is concluded that the two-dimensional homonuclear (1H,1H) and heteronuclear (1H,13C) techniques analyzed are suitable to help distinguish CBD enantiomers.

Introducción: El cannabidiol (CBD) se ha convertido en un bioactivo prometedor para las próximas décadas tras el reciente reconocimiento del potencial medicinal de los derivados del Cannabis por parte de los países miembros de las Naciones Unidas, ya que no tiene potencial psicotrópico como su isómero Δ9-tetrahidrocannabinol (Δ 9-THC). La diferenciación de estos isómeros se ha estudiado durante décadas. Estudios recientes demuestran que incluso con características químicas más sutiles, como las de los enan-tiómeros del CBD, existen diferencias bioactivas considerables. Sin embargo, no existen muchos estudios sobre sus estructuras químicas. Objetivo: Este trabajo tiene como objetivo presentar datos experimentales obtenidos por Resonancia magnética nuclear (RMN) para dilucidar mejor la estructura tridimensional de este bioactivo enantiomérico. Materiales y métodos: Para ello, una muestra de CBD no sintético de alta pureza se sometió a diferentes análisis unidimensionales (RMN-1D) y bidimensionales (RMN-2D) relacionados con los núcleos del hidrógeno (1H) y carbono (13C). Resultados y discusión: Las técnicas de RMN-1D utilizadas son suficientes para distinguir los isómeros de CBD y Δ 9-THC, pero no para identificar las características enantioméricas del CBD no sintético. Conclusiones: Se concluye que las técnicas bidimensionales homonucleares (1H,1H) y heteronucleares (1H,13C) analizadas son adecuadas para ayudar a distinguir los enantiómeros del CBD.

Introdução: O canabidiol (CBD) se tornou um bioativo promissor para as próximas décadas após o recente reconhecimento do potencial medicinal dos derivados da Cannabis pelos países membros das Nações Unidas, uma vez que não tem potencial psicotrópico como seu isômero Δ 9-tetrahidrocanabinol (A9-THC). A diferenciação desses isômeros é estudada há décadas. Estudos recentes demonstram que mesmo com características químicas mais sutis, como as dos enantiômeros do CBD, há consideráveis diferenças bioativas. Todavia, ainda não há muitos estudos sobre suas estruturas químicas. Objetivo: Este trabalho tem como objetivo apresentar dados experimentais obtidos por Ressonância magnética nuclear (RMN) para melhor elucidar a estrutura tridimensional deste bioativo enantiomérico. Materiais e métodos: Para isso, uma amostra de CBD não sintético de alta pureza foi submetida a diferentes análises unidimensionais (RMN-1D) e bidimensionais (RMN-2D) relacionadas aos núcleos de hidrogênio (1H) e carbono (13C). Resultados e discussão: As técnicas de RMN-1D usadas são suficientes para distinguir os isômeros CBD e Δ 9-THC, mas não para identificar as características enantioméricas do CBD não sintético. Conclusões: Conclui-se que as técnicas bidimensionais homonucleares (1H,1H) e heteronucleares (1H,13C) analisadas são adequadas para auxiliar na distinção dos enantiômeros do CBD.

Cloning and structural elucidation of a brassinosteroids biosynthetic gene (Atdwarf4) and genetic transformation of Indian mustard (Brassica juncea L.)

Phytohormones play critical roles in plant growth and development. Brassinosteroids (BRs) are essential group of phytohormones required for optimum growth of plants and their deficiency causes distinctive dwarf phenotypes in plants. Homeostasis of BRs in plants is maintained by DWARF4 enzyme that mediates multiple 22?-hydroxylation steps in brassinosteroid biosynthesis. Arabidopsis plants over-expressing DWARF4 show increase in inflorescence, number of branches and siliques; thereby increased number of seeds/plant. This suggests that engineering DWARF4 biosynthesis in Brassica plant can be strategized to enhance yield in mustard. In the present study (i) we cloned dwarf4 gene from Arabidopsis using gene specific PCR strategy, (ii) elucidated the three-dimensional structure of DWARF4 protein at molecular level where it revealed presence of four beta sheets and 20 alpha-helices, and (iii) transformed mustard cultivar Pusa Jaikisan with an objective to develop transgenic mustard with enhanced number of siliques. We obtained several putative transgenics with an average transformation efficiency of 3.3%. Molecular characterization with nptII specific primers confirmed presence of transgene in six putative transgenic plants.

Constructing an evaluation system for hierarchical management of clinical nurse in top three hospitals based on three-dimensional structure theory of medical quality / 中国实用护理杂志

Objective:To provide a reference for the hierarchical management of clinical nurses in top three hospitals, the hierarchical management evaluation system was conducted and evaluated in the present study.Methods:On the context of three-dimensional structure theory of medical quality, the Delphi method was applied to conduct two rounds of correspondence consultation among 23 nursing experts, and then related indexes were established based on the evaluation results and recommendations.Results:An evaluation system for hierarchical management of clinical nurse in top three hospitals was finally constructed, which included 3 first-grade and 19 second-grade indexes. The effective recovery rate of the two rounds of expert questionnaires and authority Cr of two rounds of expert consultation were 100% and 0.915, respectively. The Kendall coefficients of concordance for significance and operability of the first-grade and second-grade indexes were 0.353/0.515 and 0.344/0.469, respectively ( P<0.01). Conclusions:The hierarchical management of clinical nurses in top three hospitals constructed based on three-dimensional structure theory has reliability and feasibility.

Effect of amino acid site modification on stability of foot-and-mouth disease virus-like particles / 生物工程学报

The stability of virus-like particles (VLPs) is currently the main factor affecting the quality of foot-and-mouth disease VLPs vaccines. In order to further improve the quality of the VLPs vaccine of foot-and-mouth disease (FMD), three amino acid modification sites were designed and screened through kinetic analysis software, based on the three-dimensional structure of FMDV. The three mutant recombinant plasmids were successfully prepared by the point mutation kit, transformed into Escherichia coli strain BL21 and expressed in vitro. After purification by Ni ion chromatography column, SDS-PAGE proved that the three amino acid mutations did not affect the expression of the target protein. The results of the stability study of three FMD mutant VLPs obtained by in vitro assembly show that the introduction of internal hydrophobic side chain amino acids made the morphology of VLPs more uniform (N4017W), and their stability was significantly improved compared to the other two VLPs. The internal hydrophobic force of the capsid contributes to the formation of VLPs and helps to maintain the stability of the capsid, providing new experimental ideas for improving the quality of VLPs vaccines, and helping to promote the development of VLPs vaccines.

Sequence analysis and 3D molecular structure simulation of a novel HLA allele B*51: 159 / 中华医学遗传学杂志

Objective@#To identify a novel human leukocyte antigen (HLA) B allele in a Chinese Han individual and construct its three-dimensional structure.@*Methods@#The initial HLA genotyping was performed by PCR-sequence-based typing (PCR-SBT). The ambiguous allele was confirmed with single-strand DNA sequencing. The DNA sequence was analyzed to identify the difference between the novel allele and its closest matching allele. Finally, the three-dimensional molecular structure of the novel allele was constructed using a Swiss-Model.@*Results@#One allele of the subject at the HLA-B locus was B*44: 03: 01, whilst the other was a novel allele which differed from the closest matching allele B*51: 01: 01: 01 by nucleotide (nt) 329 A>C in exon 2, resulting in an amino acid change at codon 86 (p.Asn86Thr).@*Conclusion@#A novel HLA-B allele has been identified and officially named as HLA-B*51: 159 by the WHO Nomenclature Committee for Factors of the HLA System. The three-dimensional structure of B*51: 159 was simulated.

Structural Study on K-Segment of Rice Dehydrin in Mimetic Membrane / 分析化学

Dehydrins are well associated with the abiotic stress tolerance of the plants, such as dehydration, salt stress and cold stress. They include a highly conserved lysine-rich motif called K-segment, which is believed to play a significant role in dehydrin function. The K-segment shows in vitro antibacterial activity against Gram-positive bacteria like its full-length dehydrin protein. In this study, the structures of the K-segment from rice dehydrin have been investigated by CD spectroscopy, NMR and molecular dynamic simulation. The results reveal that the K-segment is disordered in aqueous solution, but adopts helical structure in mimetic membrane environment, sodium dodecyl sulfate (SDS) micelles. The central region of K-segment forms an a-helix and exhibits amphipathic arrangement,where hydrophobic residues locate on one side and hydrophilic residues are on the other side. The amphipathic feature allows the helix region of the K-segment to insert into the SDS micelles, resulting in stable association with mimetic membrane. To realize the energy minimization,the hydrophobic side of the helix faces to the hydrophobic core of SDS micelles,and the hydrophilic side of the helix faces to the surface of micelles. The precise 3D structure and orientation information of the K-segment obtained in this work might provide new insights in understanding the structure-function relationship of dehydrins.

Molecular characterization of matrix metalloproteinase-1 (MMP-1) in Lucilia sericata larvae for potential therapeutic applications

Background: The salivary glands of Lucilia sericata are the first organs to express specific endopeptidase enzymes. These enzymes play a central role in wound healing, and they have potential to be used therapeutically. Methods: Rapid amplification of cDNA ends and rapid amplification of genomic ends were used to identify the coding sequence of MMP-1 from L. sericata. Different segments of MMP1 gene, namely the middle part, 3' end, and 5' end, were cloned, sequenced, and analyzed using bioinformatics tools to determine the distinct features of MMP-1 protein. Results: Assembling the different segments revealed that the complete mRNA sequence of MMP-1 is 1932 bp long. CDS is 1212 bp long and is responsible for the production of MMP-1 of 404 amino acid residues with a predicted molecular weight of 45.1 kDa. The middle part, 3' end, and 5' end sequences were 933, 503, and 496 bp. In addition, it was revealed that the MMP-1 genomic sequence includes three exons and two introns. Furthermore, the three-dimensional structure of L. sericata MMP-1 protein was evaluated, and its alignment defined that it has high similarity to chain A of human MMP-2 with 100% confidence, 72% coverage, and 38% identity according to the SWISS-MODEL modeling analysis. Conclusions: MMP-1 of L. sericata has a close relationship with its homologs in invertebrates and other insects. The present study significantly contributes to understanding the function, classification, and evolution of the characterized MMP-1 from L. sericata and provides basic required information for the development of an effective medical bioproduct.

Development in research of CYP51 as the target of triazoles / 药学实践杂志

Triazoles are the most widely used antifungal drugs in clinic with broad spectrum and high efficacy,which targets sterol 14α-demethylase (CYP51),an enzyme expressed by the gene EGR11,which is a key enzyme in the fungi ergos-terol biosynthesis.On the one hand,the CYP51 belongs to a transmembrane protein.It is difficult to get the exact functional structure conformation which becomes a big challenge for the development of new drugs.On the other hand,it becomes con-sensus that EGR11 exon mutation cause CYP51 structural change is one of the major reasons for antifungal drugs resistance. Therefore,study of the structural changes toward the antifungal drug resistance is quite important.The review authors have summarized the research progress on CYP51 over the recent years.

Application of three dimensionally structured nursing management mode in the treatment of multiple trauma patients / 现代临床护理

Objective To explore the application of three dimensionally structured nursing management mode in the treatment of multiple trauma patients. Methods One hundred and eighty-seven patients with multiple injuries in the emergency department of our hospital from January to December 2010 were set up as the control group, and the traditional method was used in the group. Another 189 patients with multiple injuries from January to December 2011 were set as the observation group , in which the three dimensionally structured nursing management mode was used. The two groups were compared in terms of success rate. Result The success rates of the observation group and the control group were 85.7%(162/189) and 71.7%(134/187), respectively. The difference between the two group was statistically significant (Χ2=19.19,P<0.01). Conclusion The implementation of the three-dimensionally structured nursing management model can improve the comprehensive ability of emergency treatment and then improve the success rate of rescuing emergency patients.

Sequence analysis of a novel HLA-A?01 ∶130 allele and modeling of three-dimensional structure of HLA molecule / 中国免疫学杂志

Objective:To confirm the novel allele HLA-A*01∶130 and analyzed the nucleotide sequence of the abnormal reaction pattern.Methods: The HLA typing of sample DNA was performed by PCR-SBT.The ambiguous novel HLA allele was confirmed with single stranded SBT method,then DNA sequencing was performed to identify the difference between the novel allele and HLA-A?01:66 allele.Finally, it was modeled by Swiss-Model to three-dimensional structure of HLA Molecule.Results: The novel allele was not the same with all known HLA-A allele sequence.After analysis,there was one nucleotide differed from the A?01:66 at position 368 where A→G( codon 99 TAT→TGT) resulting in a coding change,99 Tyr was changed to Cys.The amino acid substitution at residue 99 the HLA polypeptide was located in a beta-sheet of antigenic peptide-binding region.Conclusion: The allele is a novel allele that has now been officially named as HLA-A*01∶130 by the World Health Organization( WHO) HLA Nomenclature Committee.

Allergenicity evaluation and three dimensional structure analysis of pollen panallergen Profilin from Amaranthus spinosus L / 中华微生物学和免疫学杂志

Objective To clone and characterize Profilin encoding genes in Amaranthus spinosus and to analyze the contribution of different amino acids in isoallergens to allergen antigenicity and tertiary structure. Methods The primers were designed according to the core sequences which were obtained by bioinformatic analysis of the known Profilin amino acid sequences, followed by gene cloning from the Ama- ranthus spinosus cDNA pool and subsequent confirmation by double-digestion, colony PCR and DNA sequen- cing. Antigenicity evaluation and tertiary structural modeling of the encoded protein were accomplished by online software MULTIPRED and SWISS-MODEL, respectively. Results Two panallergenic genes, named as PRF7 and PRF23, were acquired from Amaranthus spinosus. Sequence and structure analysis demonstra- ted that there was some discrepancy in tertiary structures of the encoded proteins, besides distinct difference in their amino acid sequences. PRF7 exhibited high homology with panallergen Profilins Q64LH0, with the identities 98%, whereas the homology of PRF23 and Q9XF42 (apple allergen) was 81%. Q64LH0 and PRF23 were modeled as 3nulA (Q42449) and lg5uB (Q9LE18), respectively. PRF23 exhibited distinct0 three dimensional structural difference in certain fragments compared with Q64LH0 and other Profilins. Though the average values of antigenicity displayed no difference between Q64LH0 and PRF23 on whole se- quences, the antigenicity of PRF23 on certain fragments was obviously lower than that of Q64LHO because of the alteration of some amino acids with different characters, implying the cause of lower incidence of hay fe- ver in South China than in North China. Conclusion Based on sequence analysis, antigenicity evaluation and tertiary structural modeling for Q64LH0 and PRF23, we obtained lots of useful information about the contribution of different amino acids to antigenicity and protein structures, thus would facilitate allergen ge- netic improvement by amino acid replacement.

Three Dimensional Structure of the Spinal Nerves in the Thoracolumbar Body Trunk / The Japanese Journal of Rehabilitation Medicine

The three-dimensional structure of thoracic and lumbar nerves was investigated in rats using Sihler's staining method. It was observed that the dorsal and lateral cutaneous nerves pierced the surface muscles to enter the subcutaneous tissue at a point 1-2 segments caudal to the base of the spinal nerve, while the ventral cutaneous nerves pierced the surface muscles 7 segments caudal to the base. The dorsal and lateral cutaneous nerves also ran more caudally, 2-3 and 2-4 segments, respectively, in the subcutaneous tissue. In addition, the tilt angles of spinal nerve rami lying against the transverse plane of the body trunk increased caudally, and the ventral rami showed a drastic increase in their tilt angle. Combined with our previous studies on dermatomes and sclerotomes, the present results imply that the innervation territory of the spinal nerve is shaped like a “deformed cone”, with the apex at the base of the spinal nerve and the lateral aspect expanding caudally. The stereoscopic structure of spinal nerve innervation territories may provide novel viewpoints for use in the diagnosis and treatment of disorders of the spine and spinal cord.

Micro-CT evaluation of femoral head samples' trabecular structure with femoral neck fracture / 中国矫形外科杂志

0.05).[Conclusion]The values of three-dimensional parameters in old femoral neck fracture patient are different with that of normal persons in the loading region of the femoral head although there is no difference in BMD values between them.This may provide new evidence to predict osteoporotic femoral neck fracture.

Molecular Mechanisms of Cell-cell Recognition / 生物化学与生物物理进展

Cell-cell recognition is the key for multicellular organisms to survive. This recognition critically depends on protein-protein interactions from opposing cell surfaces. Recent structural investigations reveal unique features of these cell surface receptors and how they interact. These interactions are specific, but usually relatively weak, with more hydrophilic forces involved in binding. The receptors appear to have specialized ways to present their key interacting elements for ligand-binding from the cell surface. Cell-cell contacts are multivalent. A large group of cell surface molecules are engaged in interactions. Characteristic weak interactions make possible for each individual molecule pair within the group to constantly associate-dissociate-reassociate, such that the cell-cell recognition becomes a dynamic process. The immunological synapse is a good example for immune receptors to be orchestrated in performing immunological function in a collective fashion.

Construction and Analysis of Three-dimensional Graphic Model of Single-chain Fv Derived from an Anti-human Placental Acidic Isoferritin Monoclonal Antibody by Computer / 华中科技大学学报（医学）（英德文版）

A three-dimensional (3D) graphic model of a single-chain Fv (scFv) which was derived from an anti-human placental acidic isoferritin (PAF) monoclonal antibody (Mab) was constructed by a homologous protein-predicting computer algorithm on Silicon graphic computer station.The structure, surface static electricity and hydrophobicity of scFv were investigated. Computer graphic modelling indicated that all regions of scFv including the linker, variable regions of the heavy (VH) and light (VL) chains were suitable. The VH region and the VL region were involved in composing the "hydrophobic pocket". The linker was drifted away VH and VL regions. The complementarity determining regions (CDRs) of VH and VL regions surrounded the "hydrophobic pocket". This study provides a theory basis for improving antibody affinity, investigating antibody structure and analyzing the functions of VH and VL regions in antibody activity.

Construction and Analysis of Three-dimensional Graphic Model of Single-chain Fv Derived from an Anti-human Placental Acidic Isoferritin Monoclonal Antibody by Computer / 华中科技大学学报（医学）（英德文版）

A three-dimensional (3D) graphic model of a single-chain Fv (scFv) which was derived from an anti-human placental acidic isoferritin (PAF) monoclonal antibody (Mab) was constructed by a homologous protein-predicting computer algorithm on Silicon graphic computer station.The structure, surface static electricity and hydrophobicity of scFv were investigated. Computer graphic modelling indicated that all regions of scFv including the linker, variable regions of the heavy (VH) and light (VL) chains were suitable. The VH region and the VL region were involved in composing the "hydrophobic pocket". The linker was drifted away VH and VL regions. The complementarity determining regions (CDRs) of VH and VL regions surrounded the "hydrophobic pocket". This study provides a theory basis for improving antibody affinity, investigating antibody structure and analyzing the functions of VH and VL regions in antibody activity.

Optic Disc Measurements with Stereophotograph in Normal Eyes

Optic disc can be analyzed with computerized optic nerve head analyzer or confocal scanning laser ophthalmoscope because optic disc has 3-dimensional structure. But the great cost of these instruments makes their utility for the average ophthalmologist unlikely. We measured optic discs with personal computer and stereophotograph and report as follows. Seventy-six optic discs of 76 normal subjects[61 men, 15 women; mean age: 25.5 years] were evaluated by stereoscopic optic disc photography. The area and length were measured by a computer graphic program and converted to absolute unit with Littmann's formula. The mean vertical disc diameter[1.94+/-0.24 mm], the mean horizontal disc diameter[1.67+/-0.23 mm], the mean vertical cup diameter[1.03+/-0.20 mm], the mean horizontal cup diameter[0.96+/-0.19 mm], the mean disc area[2.64+/-0.62 mm2], the mean disc circumference length[5.84+/-0.67 mm], the mean cup area[0.82+/-0.30 mm2], the mean cup base area[0.22+/-0.17 mm2], the mean cup slope area[0.60+/-0.21 mm2] and the mean rim area[1.82+/-0.41 mm2] were evaluated. The new technique to measure optic disc with stereophotograph and personal computer is thought to be a useful method to understand threedimensional structure of optic disc with relatively simple instruments.

The rapid prediction of GVHD by three-dimensional structure differences of HLA / 中国输血杂志

Objective Trying to predict the degree of GVHD after partly matched hematopoietic stem cell transplantation. Methods Analysis of the relationship between three-dimensional structure differences of donor-patient unmatched HLA and the GVHD levels after hematopoietic stem cell transplantation. Results GVHD levels were related to donor-patient unmatched HLA structure differences. The HLA structure differences forⅠ - Ⅱ degree GVHD were much smaller than that for Ⅲ - Ⅳ degree GVHD. Conclusion Prediction of GVHD by HLA structure differences is simple, rapid, specific and could help select proper conditioning regimens before transplantation and the proper immune suppressive agents after transplantation.

THE APPLICATION OF MOLECULAR MODELING OF HLA ANTIGENS IN UNMATCHED STEM CELL TRANSPLANTATION FOR GVHD PREDICTION / 解放军医学杂志

To explore the pathogenic mechanism of GVHD, attempting to forecast the degree of GVHD after stem cell transplantation, and to enhance the therapeutic effectiveness of clinical transplantation. The conventional microlymphocytotoxicity and sequencing methods were used in typing the HLA. The degrees of GVHD were estimated by molecular modeling of HLA antigens and veryfied the estimation by comparing the clinical results with anticipated degrees. In 8 recipients, three were transplanted with half matched stem cells. Among these 3 pationts, two developed IV degree GVHD, and one developed II degree GVHD. In the other 3 patients, cells of two unmatched HLA antigens were transplanted, and among them one developed I degree GVHD, and two developed II degree GVHD. In two patients who were transplanted with cells of one unmatched HLA antigens, I and II degrees GVHD developed respectively. Second, the correlationship analysis showed that degrees of GVHD had positive correlation with the RMSD (relative mean square deviation) between different HLA antigens. These results indicated that the degrees of GVHD after stem cell transplantation were related with the difference of three dimensional structures of unmatched stem cell HLA antigens; molecular modeling might be used to predict the prognosis of clinical stem cell transplantation.