Matrix Protein Gene Research Articles

Effects of nicotine on porcine pre-pupertal sertoli cells: An in vitro study

Smoke components, such as nicotine and its major metabolites, cross the blood–testis barrier and are detectable in the seminal plasma of both active smokers and individuals exposed to cigarette smoke. In vivo studies in a rat model have further demonstrated that nicotine exposure reduces the weight of the testis, as well as the number of spermatocytes and spermatids, and affects the ultrastructure of Sertoli cells (SC) – which serve as sentinels of spermatogenesis - causing intense germ cell sloughing in the tubular lumen that compromises offspring fertility.This study sought to determine the effects of nicotine on the viability and function of purified pig pre-pubertal SC. Nicotine exposure reduced the mRNA expression and protein levels of anti-Mullerian hormone (AMH) and inhibin B and impaired FSH-r sensitivity via the downregulation of FSH-r and aromatase gene expression compared to untreated SC. Overall, our study suggests that nicotine can significantly alter extracellular matrix and tight junction protein gene expression (e.g., laminin, integrin, and occludin), thus compromising cross-talk between the interstitial and tubular compartments and enhancing blood–testis barrier (BTB) permeability via downregulation of the mitogen-activated protein kinase (MAPK) pathway.These findings further elucidate a potential mechanism of action underlying nicotine exposure's detrimental effects on SC function in vivo.

Expression of Two Foreign Genes by a Newcastle Disease Virus Vector From the Optimal Insertion Sites through a Combination of the ITU and IRES-Dependent Expression Approaches.

Many Newcastle disease virus (NDV) strains have been developed as vectors to express a foreign gene (FG) for vaccine and cancer therapy purposes. The non-coding region between the phosphoprotein (P) and matrix protein (M) genes and the non-coding region behind the NP gene open reading frame (ORF) in the NDV genome have been identified as the optimal insertion sites for efficient FG expression through the independent transcription unit (ITU) and the internal ribosomal entry site (IRES) dependent expression approaches, respectively. To date, however, the majority of these NDV vectors express only a single or two FGs from suboptimal insertion sites in the NDV genome, obtaining various levels of FG expression. To improve the FG expression, we generated NDV LaSota vaccine strain-based recombinant viruses expressing two FGs, GFP, and RFP, from the identified optimal insertion sites through a combination of the ITU and IRES-dependent approaches. Biological assessments of the recombinant viruses indicated that the recombinants expressing two FGs were slightly attenuated with approximately one order of magnitude lower in virus titers when compared to the viruses containing a single FG. The FG expression efficiencies from the two-FG viruses were also lower than those from the single-FG viruses. However, the expression of two FGs from the optimal insertion sites was significantly (p < 0.05) higher than those from the suboptimal insertion sites. The expressions of FGs as monocistronic ITU were approximately 4-fold more efficient than those expressed by the bicistronic IRES-dependent approach. These results suggest that the NDV LaSota vector could efficiently express two FGs from the identified optimal insertions sites. The ITU strategy could be used for “vectoring” FGs in circumstances where high expression of gene products (e.g., antigens) is warranted, whereas, the IRES-dependent tactic might be useful when lower amounts of IRES-directed FG products are needed.

A CTGF-YAP Regulatory Pathway is Essential for Angiogenesis and Barriergenesis in the Retina

Connective tissue growth factor (CTGF) or CCN2 is a matricellular protein essential for normal embryonic development and tissue repair. CTGF exhibits cell- and context-dependent activities, but the CTGF function in vascular development and permeability barrier is not known. Here we show that endothelial cells (ECs) are one of the major cellular sources of CTGF in the developing and adult retinal vasculature. Mice lacking CTGF expression either globally or specifically in ECs exhibit impaired vascular cell growth and morphogenesis, and blood barrier breakdown. The global molecular signature of CTGF includes cytoskeletal and extracellular matrix protein, growth factor, and transcriptional co-regulator genes such as yes-associated protein (YAP). YAP, itself a transcriptional activator of the CTGF gene, mediates several CTGF-controlled angiogenic and barriergenic transcriptional programs. Re-expression of YAP rescues, at least partially, angiogenesis and barriergenesis in CTGF mutant mouse retinas. Thus, the CTGF-YAP angiomodulatory pathway is critical for vascular development and barrier function.

Whole-genome next-generation sequencing and phylogenetic characterization of viral haemorrhagic septicaemia virus in Korea.

Whole-genome next-generation sequencing was used to investigate the local evolution of viral haemorrhagic septicaemia virus, a serious pathogen affecting economically important fish such as rainbow trout and turbot in Europe and olive flounder in Asia. Sequence analysis showed that all isolates were genotype IVa, but could be classified further into four subgroups (K1-K4). In addition, genomic regions encompassing the nucleoprotein, phosphoprotein, matrix protein and non-virion protein genes, as well as the seven non-coding regions, were relatively conserved, whereas glycoprotein and RNA-dependent RNA polymerase genes were variable in the coding region. Taken together, the data demonstrate that whole-genome next-generation sequencing may be useful for future surveillance, prevention and control strategies against viral haemorrhagic septicaemia.

Clinical characteristics and pathogenic gene analysis in a large pedigree with multiple epiphyseal dysplasia

Objective To provide experimental evidence for genetic counseling and prenatal molecular diagnosis by analyzing the clinical characteristics and screening for pathogenic genes of a five-generation suspected multiple epiphyseal dysplasia (MED) family (17 patients). Methods The family members' medical history, general physical examination and hip joint X-ray examination were collected. Peripheral blood samples of the family members were collected and DNA were extracted from these samples. The exons of clinical genes from probands' DNA were sequenced by High throughput sequencing method. Next Gene software was used to compare and analyze the sequence and INGENUITY software was further used to annotate the mutations in order to find the pathogenic mutations in probands. The suspicious mutations were confirmed in pedigree members by PCR and Sanger sequencing. Results The family consisted of 5 generations and 38 members. Pedigree analysis was consistent with autosomal dominant inheritance. There were 17 patients in the family, and their clinical manifestations showed abnormal walking posture in childhood, pain in hip and knee joints, and typical pathological changes of epiphyseal dysplasia on X-ray. Cartilage oligomeric matrix protein (COMP) gene c.1153G>A (p.Asp385Asn) missense heterozygous mutation was screened in proband, which was genotypically and phenotypically segregated in the pedigree. Conclusion A missense mutation of the comp gene has been identified in a pedigree affected with MED which was the first reported in a big family. Our result is conducive to the further diagnosis and treatment and also provides a molecular basisfor the future prenatal diagnosis. Key words: Osteochondrodysplasias; Pedigree; DNA, intergenic; DNA mutational analysis

Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria

Background and Aim:Peste des petits ruminants (PPR) is an acute, extremely contagious transboundary viral disease of small ruminants with severe economic consequences, caused by PPR virus. Cost-effective and rapid diagnosis of the disease is essential for prompt management and control. This study aimed to compare the application of a commercial colorimetric loop-mediated isothermal amplification (cLAMP) kit and reverse transcriptase-polymerase chain reaction (RT-PCR) in the diagnosis of PPR in sheep and goats in Southeast Nigeria.Materials and Methods:Nasal swab samples were collected from West African Dwarf sheep and goats showing clinical signs suggestive of PPR (n=80) and those without any clinical signs (n=140) of the disease. The diagnosis was achieved through detection of PPR viral genome in the samples using a cLAMP kit and RT-PCR. cLAMP assay was done directly on nasal swab samples without ribosomal nucleic acid extraction. A set of six primers targeting the matrix gene protein was used for the cLAMP assay.Results:PPR viral genome was detected by both cLAMP and RT-PCR in 51 (63.8%) of the 80 samples from sheep and goats with signs suggestive of PPR while 14 (10%) of those without signs tested positive for PPR by both assay methods. There was a 100% agreement in the cLAMP and RT-PCR results. However, cLAMP was a faster, easier, and less expensive method compared to RT-PCR.Conclusion:The cLAMP assay demonstrates the potential for a point of care diagnosis in the field and a valuable diagnostic tool in areas with poor electricity supply as well as in a less equipped diagnostic laboratory. Since the reagents are affordable, cLAMP can be a diagnostic tool of choice in the detection and surveillance of PPR virus in countries with limited resources.

Clinical characteristics of pseudoachondroplasia and analysis of COMP gene mutation

Objective This article reported the clinical characteristics and gene mutations of two pseudoachondroplasia cases, and made a literature review in order to improve clinicians′ understanding of the disease. Methods Clinical features of two patients who were short stature accompanied with skeletal deformities were summarized, and they accepted whole exome sequencing. We also reviewed literature to summarize the clinical characteristics and known gene research progress of all reported Chinese pseudoachondroplasia cases. Results The two patients′ clinical characteristics were short limbdwarfism with skeletal deformity. Genetic results showed that there were two heterozygous mutations in the cartilage oligomeric matrix protein (COMP) gene of the two patients, c. 1417_1419delGAC and c. 1552G>A, respectively. Up to March 2019, a total of 58 cases of pseudoachondroplasia have been reported in China. The median height of these patients is -5.03 SDS. The clinical features include abnormal gait, short limbs, short fingers/toes, scoliosis, bracelet sign, ankle sign and other skeletal deformities. COMP is the pathogenic gene and mutations mainly located in calmodulin-like domains. The hotspot mutation is c. 1417_1419delGAC. Conclusions Pseudoachondroplasia is a kind of rare genetic disease characterized by short stature and skeletal deformities. The clinical and genetic characteristics of the disease were summarized, which may improve the early diagnosis rate. Key words: Pseudoachondroplasia; Cartilage oligomeric matrix protein; Gene mutation; Molecular mechanism

Recombinase polymerase amplification–nucleic acid lateral flow immunoassays for Newcastle disease virus and infectious bronchitis virus detection

Newcastle disease virus (NDV) and infectious bronchitis virus (IBV) are two poultry pathogens affecting the respiratory tract of chickens, and cause major economic losses in the industry. Rapid detection of these viruses is crucial to inform implementation of appropriate control measures. The objective of our study is developing a simple, rapid and field applicable recombinase polymerase amplification (RPA)–nucleic acid lateral flow (NALF) immunoassay for detection of NDV and IBV. Isothermal amplification of the matrix protein (M) gene of NDV and the nucleoprotein (N) gene of IBV was implemented via recombinase polymerase amplification at 38 °C for 40 min and 20 min, respectively using modified labeled primers. NALF device used in this study utilizes antibodies for detection of labeled RPA amplicons. The results revealed that RPA-NALF immunoassays can detect both viruses after 40 min at 38 °C and only NDV after 20 min. The limit of detection (LOD) was 10 genomic copies/RPA reaction. The assays results on clinical samples collected from diseased chicken farms demonstrated a good performance in comparison with quantitative real time reverse transcription polymerase chain reaction (qRT-PCR). The assays established in this study can facilitate rapid, on-site molecular diagnosis of suspected cases of ND and IB viral infections as the results can be detected by the naked eye without the need for measuring fluorescence. Furthermore, the NALF device could be adapted to detect other infectious agents.

Highly Pathogenic Avian Influenza H5N1 (HPAI/H5N1) Virus Search from Wild Birds in Ghana

Abstract Highly pathogenic avian influenza (AI) disease has occurred in many countries globally adversely affecting domestic poultry production. Ghana recorded her first outbreak of a highly pathogenic avian influenza (HPAI) in 2007 on a small scale commercial farm in Tema. Since then, there have been numerous outbreaks. The source of these outbreaks is not conclusive. The role of wild birds in the epidemiology of avian influenza outbreaks in Ghana is not known. This study sought to investigate the role of wild birds in the outbreaks of Highly Pathogenic Avian Influenza (HPAI H5N1) in Ghana, particularly in Southern Ghana. Wild birds were trapped and sampled through mist netting. The faecal and tracheal samples were analysed using a One-Step Real Time Reverse Transcription Polymerase Chain reaction (RT-PCR) with primer sets targeting the matrix protein gene of the Avian influenza virus. Sera samples were subjected to multispecies competitive Enzyme Linked Immunosorbent Assay (ELISA) for anti-AI virus antibodies. Three hundred and twenty two (322) wild birds were trapped and sampled. Birds sampled included 87.3 % (281/322) resident birds and 12.7 % (41/322) migratory birds. The migratory birds included intra-African migrants 12.2 % (5/41) and Pale-arctic migrants 87.8 % (36/41). Avian influenza virus and antibody were neither detected in these swabs nor sera samples, respectively. The study documented the absence of AI in resident and migrant wild birds in the study area and suggest that wild birds may not be responsible for the outbreaks of AI in the poultry. However, sustained surveillance is recommended to ascertain a nationwide successful prevention and control strategy to stay the tide of any future intruding AI outbreaks.

The Nucleoprotein and Phosphoprotein Are Major Determinants of the Virulence of Viral Hemorrhagic Septicemia Virus in Rainbow Trout.

Viral hemorrhagic septicemia virus (VHSV), a fish rhabdovirus, infects several marine and freshwater fish species. There are many strains of VHSV that affect different fish, but some strains of one genetic subgroup have gained high virulence in rainbow trout (Oncorhynchus mykiss). To define the genetic basis of high virulence in trout, we used reverse genetics to create chimeric VHSVs in which viral nucleoprotein (N), P (phosphoprotein), or M (matrix protein) genes, or the N and P genes, were exchanged between a trout-virulent European VHSV strain (DK-3592B) and a trout-avirulent North American VHSV strain (MI03). Testing of the chimeric recombinant VHSV (rVHSV) by intraperitoneal injection in juvenile rainbow trout showed that exchanges of the viral P or M genes had no effect on the trout virulence phenotype of either parental strain. However, reciprocal exchanges of the viral N gene resulted in a partial gain of function in the chimeric trout-avirulent strain (22% mortality) and complete loss of virulence for the chimeric trout-virulent strain (2% mortality). Reciprocal exchanges of both the N and P genes together resulted in complete gain of function in the chimeric avirulent strain (82% mortality), again with complete loss of virulence in the chimeric trout-virulent strain (0% mortality). Thus, the VHSV N gene contains an essential determinant of trout virulence that is strongly enhanced by the viral P gene. We hypothesize that the host-specific virulence mechanism may involve increased efficiency of the viral polymerase complex when the N and P proteins have adapted to more efficient interaction with a host component from rainbow trout.IMPORTANCE Rainbow trout farming is a major food source industry worldwide that has suffered great economic losses due to host jumps of fish rhabdovirus pathogens, followed by evolution of dramatic increases in trout-specific virulence. However, the genetic determinants of host jumps and increased virulence in rainbow trout are unknown for any fish rhabdovirus. Previous attempts to identify the viral genes containing trout virulence determinants of viral hemorrhagic septicemia virus (VHSV) have not been successful. We show here that, somewhat surprisingly, the viral nucleocapsid (N) and phosphoprotein (P) genes together contain the determinants responsible for trout virulence in VHSV. This suggests a novel host-specific virulence mechanism involving the viral polymerase and a host component. This differs from the known virulence mechanisms of mammalian rhabdoviruses based on the viral P or M (matrix) protein.

Orthopaedic manifestations of pseudoachondroplasia.

PurposeIn 1959, Maroteaux and Lamy initially designated pseudoachondroplasia as a distinct dysplasia different from achondroplasia the most common form of skeletal dysplasia. Pseudoachondroplasia is caused by a mutation in the collagen oligomeric matrix protein gene (COMP) gene on chromosome 19p13.1-p12 encoding the COMP. The COMP gene mutations result in rendering the articular and growth plate cartilages incapable of withstanding routine biomechanical loads with resultant deformity of the joints. The purpose of the study was to characterize the typical orthopaedic findings in pseudoachondroplasia.MethodsThe charts and radiographs of 141 patients with pseudoachondroplasia were analyzed. This cohort, to our knowledge, represents the largest group of patients describing the typical orthopaedic manifestations of pseudoachondroplasia.ResultsPatients with pseudoachondroplasia have normal craniofacial appearance with normal intelligence. Short stature is not present at birth and generally appears by two to four years of age. The condition is a form of spondyloepiphyseal dysplasia and the long bones are characterized by dysplastic changes in the epiphysis, metaphysis and vertebral bodies. Radiographically the long bones have altered the appearance and structure of the epiphyses with small irregularly formed or fragmented epiphyses or flattening. The metaphyseal regions of the long bones show flaring, widening or ‘trumpeting’. The cervical (89%) and thoracic and lumbar vertebrae show either platyspondyly, ovoid, ‘cod-fish’ deformity or anterior ‘beaking’. Kyphosis (28%), scoliosis (58%) and lumbar lordosis (100%) are commonly seen. The femoral head and acetabulum are severely dysplastic (100%). The knees show either genu valgum (22%), genu varum (56%) or ‘windswept’ deformity (22%).ConclusionMost commonly these distortions of the appendicular and the axial skeleton lead to premature arthritis particularly of the hips and often the knees not uncommonly in the 20- to 30-year-old age group.Level of Evidence:III

Identification of nacre matrix protein genes hic14 and hic19 and their roles in crystal growth and pearl formation in the mussel Hyriopsis cumingii.

Biological mineralization is a highly programmed process in which inorganic minerals reassociate under the strict control of the extracellular matrix to form minerals with special functions and patterns. Shells are biominerals, and their synthesis is finely regulated by organic matrix including matrix proteins, polysaccharides, lipids, pigments, free amino acids, and small peptides. In this study, two matrix protein genes, hic14 and hic19, were isolated from the mantle of the mussel Hyriopsis cumingii. Tissue expression analysis showed that both proteins were expressed mainly in the mantle, and in situ hybridization of mantle tissues showed that they were specifically expressed in the dorsal epithelial cells of mantle pallial. Therefore, hic14 and hic19 were both nacreous layer matrix proteins. In the pearl insertion experiment, hic14 and hic19 kept low expression during pearl sac formation and disordered calcium carbonate deposition, and increased significantly during pearl nacre accumulation, which showed that both proteins participated in the mineralization of pearl nacre. In the RNA interference experiment, shell nacre tablet growth was inhibited after crystal nucleation due to the decreased expression of hic14, and crystal morphology and arrangement of nacre were highly modified after expression of hic19 was inhibited. These results provided further evidence that both hic14 and hic19 participated in nacreous layer biomineralization.

Hichin, a chitin binding protein is essential for the self-assembly of organic frameworks and calcium carbonate during shell formation

Shell biomineralization is a process where inorganic minerals accumulate upon a chitinous scaffold under the control of multifunctional matrix proteins. In this study, we cloned a novel matrix protein gene from the mantle of Hyriopsis cumingii. The predicted protein, hichin, contains a chitin-binding domain and exhibited the highest expressional level in mantle tissue, with positive signals mainly detected in dorsal epithelial cells of the pallial mantle according to in situ hybridization, indicating its possible involvement in shell nacreous layer biomineralization. RNA interference showed that hichin suppression induced disordered self-assembly of the insoluble framework in the nacreous layer, and that the newly formed calcium carbonate crystals could not bind to organic frameworks. Furthermore, hichin was primarily responsible for building the framework during initial nacre deposition in pearl formation. Moreover, the chitin-binding domain of hichin also provided crystal morphology regulation in vitro crystallization assay. These results indicated that hichin is involved in the self-assembly of organic frameworks and morphological regulation in shell nacreous layer.

The global effects of PmRunt co-located and co-expressed with a lincRNA lncRunt in pearl oyster Pinctada fucata martensii

Runt related transcription factors as trans-acting elements play critical roles in the developmental control of cell fate, hematopoiesis, bone formation and cancers. In previous study, the homologue of runt related transcription factor PmRunt has been identified from pearl oyster Pinctada fucata martensii and considered to play an important role in nacre formation. In this study, we used the same samples to perform RNA-seq to detect the global effects after the decrease of PmRunt expression. The transcription levels of several nacre shell matrix protein (NSMP) genes were significantly changed and the potential compensatory effect could happen internal gene families. Downregulation of PmRunt could also influence the biosynthesis of NSMPs through affecting amino acid metabolism, translation, protein processing and export. The inhibition of PmRunt also possibly affected the expression of caspases, IAPs and C1qs that related to apoptosis and immune. In addition, PmRunt highly expressed at 12 h and 12 d after transplantation in hemolymph, which was corresponded to transplantation immunity immune response and the morphology of pearl sac, suggested the cross-talk of biomineralization-immune regulation in hemocytes. Furthermore, a lincRNA (LncRunt) that co-located with PmRunt was identified and showed a significantly relative expression with PmRunt, which suggested the potential regulation. Therefore, these findings provided new idea to find the regulation targets of runt-related transcription factors and offers evidence of lncRNAs in potential biomineralization-immune regulation.

Effects of magnesium deprivation on development and biomineralization in the sea urchin Arbacia lixula

ABSTRACTEchinoderms have an extensive endoskeleton composed of magnesian calcite and occluded matrix proteins. As biomineralization in sea urchin larvae is sensitive to the Magnesium:Calcium ratio of seawater, we investigated the effects of magnesium deprivation on development and skeletogenesis in the sea urchin Arbacia lixula. We focused on the localization of the skeletogenic cells (primary mesenchyme cells) and the spatial expression of associated genes. Embryos reared in Mg-free seawater exhibited developmental delay from 6-h post-fertilization and at 24 h embryos showed complete lack of biomineral formation. Larvae (48–72 h) exhibited severe skeleton malformations. Fluorescent labelling revealed that the primary mesenchyme cells and the developing skeleton of treated embryos were in an abnormal ectopic location. Expression of the skeleton matrix protein gene (msp130) in the primary mesenchyme cells as seen using in situ hybridization was normal at 24 h. At 48 h this gene was down-regulated in control larvae, but not in treated larvae. Development of the pigment cells, immune cells that, like the skeleton, are mesodermal derivatives, was also impaired. Our results highlight the essential role of Mg in skeleton formation in sea urchin embryos with an indication that this element is also generally important for the development of mesoderm.Abbreviations: hpf: hours post fertilization; PMCs: primary mesenchyme cells; ACC: amorphous calcium carbonate; MgFSW: magnesium-free seawater; FSW: filtered seawaterImpact statementEchinoderms have an extensive endoskeleton composed of magnesian calciteWe investigated the effects of magnesium deprivation on development and skeletogenesis in the sea urchin Arbacia lixulaMagnesium deprivation caused developmental delay and skeleton malformationsPrimary mesenchyme cells of treated embryos were in an abnormal ectopic locationThe spatial and temporal expression profile of the skeleton matrix protein gene (msp130) was found to be different from controlsOur results highlight the essential role of Mg across developmental processes in sea urchin embryos.

Regulation of Proliferation, Differentiation and Functions of Osteoblasts by Runx2.

Runx2 is essential for osteoblast differentiation and chondrocyte maturation. During osteoblast differentiation, Runx2 is weakly expressed in uncommitted mesenchymal cells, and its expression is upregulated in preosteoblasts, reaches the maximal level in immature osteoblasts, and is down-regulated in mature osteoblasts. Runx2 enhances the proliferation of osteoblast progenitors by directly regulating Fgfr2 and Fgfr3. Runx2 enhances the proliferation of suture mesenchymal cells and induces their commitment into osteoblast lineage cells through the direct regulation of hedgehog (Ihh, Gli1, and Ptch1), Fgf (Fgfr2 and Fgfr3), Wnt (Tcf7, Wnt10b, and Wnt1), and Pthlh (Pthr1) signaling pathway genes, and Dlx5. Runx2 heterozygous mutation causes open fontanelle and sutures because more than half of the Runx2 gene dosage is required for the induction of these genes in suture mesenchymal cells. Runx2 regulates the proliferation of osteoblast progenitors and their differentiation into osteoblasts via reciprocal regulation with hedgehog, Fgf, Wnt, and Pthlh signaling molecules, and transcription factors, including Dlx5 and Sp7. Runx2 induces the expression of major bone matrix protein genes, including Col1a1, Spp1, Ibsp, Bglap2, and Fn1, in vitro. However, the functions of Runx2 in differentiated osteoblasts in the expression of these genes in vivo require further investigation.

Transcription factor Pf-Rel regulates expression of matrix protein genes Prismalin-14 and MSI60 in the pearl oyster Pinctada fucata.

Molluscan shell is a biomineral that consists of a highly organized calcium carbonate composite. Organisms mainly use matrix proteins to elaborately control the biomineralization process, but knowledge of their regulatory mechanisms is limited. The transcription factor Pf-Rel, which belongs to the Rel/nuclear factor-κB family, was shown to regulate transcription at the Nacrein promoter in the pearl oyster Pinctada fucata. Here, we further explored the transcriptional regulation mechanisms of Pf-Rel on the matrix proteins Prismalin-14 and MSI60. The relative expression levels of Prismalin-14 and MSI60 were high in the mantle edge and mantle pallial tissues of P. fucata. These three genes were significantly up-regulated after shell notching, suggesting that they might play important roles during shell formation. Importantly, Pf-Rel gene knockdown by RNA interference led to down-regulation of Prismalin-14 and MSI60 expression. In transient co-transfection assays, Pf-Rel significantly up-regulated the promoter activities of the Prismalin-14 and MSI60 genes in a dose-dependent manner. Furthermore, the promoter regions of Prismalin-14 (-1794 to -1599bp) and MSI60 (-2244 to -1141bp) were required for the activation by Pf-Rel. Altogether, these results suggest that the transcription factor Pf-Rel can up-regulate the expression of the matrix protein genes Prismalin-14 and MSI60 during shell formation in P. fucata, which improves our understanding of transcription regulation at the molecular level during molluscan shell development.

Anti-colon cancer effect of matrix protein gene therapy with nanoparticles

The matrix protein (MP) is the core structure of the oncolytic vesicular stomatitis virus (VSV) and it played a vital role during the process of VSV infection and corresponding cytopathic effects in cancer treatment. In this study, a novel delivery synthesized by PEG-PCL-PEG and DOTAP (PPPD) was applied to encapsulate MP plasmids to derive the advantage and overcome the drawback of VSV platform in the treatment of colon cancer. The MP/DPPP micelles were stable in physiological solution, with an average diameter of 102 nm and zeta potential of 5.4 mV. This delivery system has a high encapsulation rate of 96% and transfection efficiency of 36%. A prominent anti-cancer activity of MP/DPPP was found both in Ct26 cells and animal models. The mechanisms of antitumor were further explored in this research. Our results indicated that MP/DPPP micelles were promising agents for future clinical application in the treatment of colon cancer.

Clinical features and gene mutation analysis of COMP in eight patients with pseudoachondroplasia

The clinical features and the mutation of cartilage oligomer matrix protein (COMP) gene were analyzed in 8 patients with pseudoachondroplasia (PSACH). The clinical data and the peripheral blood from 5 male and 3 female probands, their pedigree members, and 250 unrelated volunteers were collected. Eight patients who were sporadic cases, had been detected mutation of COMP gene by DNA sequencing. PSACH is a skeletal disorder characterized by short stature, joint laxity, and early-onset osteoarthritis. The heights of 8 patients were significantly lower than the average level by 3 standard deviations, with short limbs and deformities of legs. Radiographs showed flattening of vertebrae with anterior beaking or tonguing in children and osteoarthritis in adults. As to the patients with short limb dwarfism, short toes, and abnormal radiography findings, PSACH should be suspected and could be confirmed by detection of COMP gene mutation. Key words: Pseudoachondroplasia; Clinical features; Cartilage oligomer matrix protein; Gene mutation

Expressions of Shell Matrix Protein Genes in the Pearl Sac and Its Correlation with Pearl Weight in the First 6Months of Pearl Formation in Hyriopsis cumingii.

Matrix proteins regulate crystal nucleation, morphology, and polymorphism during pearl biomineralization and have significant correlations with pearl quality traits in nucleated pearls. However, there is little information about the connection between pearl quality traits and matrix proteins in non-nucleated pearls. In this study, we analyzed CaCO3 deposition during the first month of non-nucleated pearl formation and examined the expression patterns of ten shell matrix protein genes (Hcperlucin, hic31, silkmapin, hic22, hic74, hic52, HcTyr, HcCA3, hic24 and Hc-upsalin) in the pearl sac of Hyriopsis cumingii. During pearl formation, CaCO3 crystals were initially deposited in a disorderly manner during days 12 and 15 of pearl formation. On days 18 and 21, CaCO3 crystals gradually nucleated on an organic membrane, and the pattern of crystal deposition changed markedly. Between days 24 and 30, crystals similar to nacre tablets were deposited; they then grew and formed connections in a more orderly fashion, eventually forming the nacreous layer. We observed high expression levels of shell matrix proteins during the phases of disordered or ordered CaCO3 deposition, suggesting they were involved in non-nucleated pearl formation. Furthermore, the expressions of nine matrix proteins were significantly correlated with pearl weight during the first 6months after grafting. The prismatic-layer matrix protein hic31 and nacreous-layer matrix protein hic22 showed negative correlations with pearl weight, but the other seven nacreous-layer matrix proteins had significantly positive correlations with pearl weight. These results show the involvement of matrix proteins in pearl formation and in determination of quality traits.