Objective Two autosomal recessive forms of muscular dystrophy:LGMD2B and Miyoshi myopathy may be indused by dysferlin gene mutation.The purpose of this study was to define molecular defects in dysferlin gene in a family with Miyoshi myopathy.Methods mRNA from peripheral blood in a Chinese Miyoshi myopathy pedigree was amplified by RT-PCR and the mutation was determined by sequencing the amplified products.Results The results of sequencing revealed a novel homozygous mutation,a 6429delG,on exon 53 of the dysferlin gene for the patients.Conclusion The 6429delG mutation in the dysferlin gene of patients creates a frameshift mutation which induces a stop codon at 2035 on exon 54 and the premature dysferlin contributes to the Miyoshi myopathy in the Chinese pedigree.

Objective To study the anticoagulant effects of Pheretima extracts and analyze the effective constituents. Methods Antithrombotic effects of Pheretima extracts were observed by determining the coagulation time, bleeding time of mice and the recalcification time of rabbit plasma in vitro, meanwhile the content of free amino acid in the extracts were determined. Results Pheretima extracts could prolong the coagulation time, bleeding time of mice, and the recalcification time of rabbit plasma in vitro. The content of free amino acid was rich in Pheretima extracts. Conclusion Free amino acid in Pheretima extracts may be the main anticoagulant effective constituents.

Objective To establish method for the fingerprint of Radix et Rhizoma Gentianae using UFLC with the hierarchical cluster analysis.Methods Column:Shimadzu C18(3.0 mm?75 mm,1.7 ?m);mobile phase:Acetonitrile-0.4% phosphorylation(5.5∶94.5);flow rate:0.8 mL/min;detection wavelength:240 nm;temperature:40 ℃.Results This method had a good linearity in the range of 0.154 8—1.858 mg/mL,and the average recovery was 102.09% with RSD of 1.68% for swertiamarin.It had a good linearity in the range of 0.322—5.152 mg/mL,and the average recovery was 103.80% with RSD of 1.11% for gentiopicroside.The RSD of precision and reproducibility were lower than 2.1%.The result of fingerprint studying was the same with the hierarchical cluster analysis.Conclusion This method can be used as standardized implantation and quality control of Radix et Rhizoma Gentianae.

OBJECTIVETo identify an inbred Chinese pedigree with autosomal recessive muscular dystrophy and analyze the molecular defects.

METHODSLinkage analysis was conducted using short tandem repeat(STR) markers from the regions associated with limb-girdle muscular dystrophy type 2A(LGMD2A) through 2H. Multi-Western blot was performed with anti-calpain-3, anti-dysferlin, anti-gamma-sarcoglycan, anti-alpha-sarcoglycan, and anti-dystrophin monoclonal antibodies. Mutation was determined by reverse transcriptase-polymerase chain reaction and sequencing.

RESULTSTwo-point linkage analysis showed significant Lod scores with markers from chromosome 2p13, the highest two-point Lod scores were obtained with D2S337 (Z(max)=1.86 at theta=0). Multi-Western blot confirmed dysferlin deficiency of muscle specimen from the proband. Mutation analysis revealed a novel 6429delG mutation on exon 53 of the DYSF gene for the proband.

CONCLUSIONThe authors identified an inbred Chinese pedigree with Miyoshi myopathy caused by a 6429delG on the DYSF gene. This mutation is predicted to result in premature termination of translation.

DNA, Complementary ; chemistry ; Dysferlin ; Genetic Linkage ; Humans ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Muscle Proteins ; genetics ; Muscular Diseases ; genetics ; Muscular Dystrophies ; genetics ; Mutation ; Pedigree

DNA barcoding is a powerful approach for characterizing species of organisms, especially those with almost identical morphological features, thereby helping to to establish phylogenetic relationships and reveal evolutionary histories. In this study, we chose a 648-bp segment of the mitochondrial gene, cytochrome c oxidase subunit 1 (COI), as a standard barcode region to establish phylogenetic relationships among brine shrimp (Artemia) species from major habitats around the world and further focused on the biodiversity of Artemia species in China, especially in the Tibetan Plateau. Samples from five major salt lakes of the Tibetan Plateau located at altitudes over 4,000 m showed clear differences from other Artemia populations in China. We also observed two consistent amino acid changes, 153A/V and 183L/F, in the COI gene between the high and low altitude species in China. Moreover, indels in the COI sequence were identified in cyst and adult samples unique to the Co Qen population from the Tibetan Plateau, demonstrating the need for additional investigations of the mitochondrial genome among Tibetan Artemia populations.
Animals ; Artemia ; classification ; genetics ; Base Sequence ; China ; DNA, Mitochondrial ; chemistry ; genetics ; Electron Transport Complex IV ; genetics ; Genetic Variation ; Molecular Sequence Data ; Phylogeny ; Selection, Genetic ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid ; Tibet

Accurate classification of the acetabular injuries and appropriate treatment plan are great challenges for orthopedic surgeons because of the irregular anatomical structure of the acetabulum and aggregation of important vessels and nerves around it. Letournel-Judet classification system has been widely applied to classify acetabular fractures. However, there are several limitations, including incomplete inclusion of fracture types, difficulty in understanding and insufficient guidance for surgical treatment, etc. Serious complications such as traumatic arthritis are common due to wrong classification and diagnosis and improper selection of surgical strategy, which brings a heavy burden to the society and families. Three-column classification, based on anatomic characteristics, has advantages of containing more fracture types and being easy to understand, etc. To solve the problems existing in the diagnosis and treatment process based on Letournel-Judet classification, achieve accurate diagnosis and treatment of patients with acetabular fractures, and obtain satisfactory prognosis, the Orthopedic Trauma Emergency Center of Third Hospital of Hebei Medical University and the Trauma Orthopedic Branch of the Chinese Orthopedic Association organized experts from relevant fields to formulate the Expert consensus on the accurate diagnosis and treatment of acetabular fractures based on three-column classification ( version 2023) in terms of principles of evidence-based medicine. Based on the three-column classification, 15 recommendations were proposed, covering the diagnosis, treatment, complication prevention and management, etc, so as to provide reference for accurate diagnosis and treatment of acetabular fractures.

OBJECTIVE： To investigate the effects and its mechanism of calcium phosphate bone cement （CPC） loading total flavonoids of Davallia mariesii on osteogenic differentiation of induced membrane in rats. METHODS： Drug-loading CPC and drug-loading polymethyl methacrylate （PMMA） cement were prepared with the contents of Qianggu capsules （total flavonoids of D. mariesii as active ingredient） using CPC and PMMA cement as carrier. Totally 64 male SD rats were randomly divided into drug-loading CPC group， drug-loading PMMA cement group， no-drug CPC group， no-drug PMMA cement group， with 16 rats in each group. The femur of rats was separated and osteotomized to prepare bone defect model， and then the corresponding bone cement was implanted. Four weeks after modeling， the induced membranes of rats were cut and protected. Bone cement was taken out and autogenous cancellous bone was implanted. At the 4th week after modeling， X-ray photographs were taken on the hind limb bones of rats. At the 4th week after modeling and 6th week after bone grafting， induced membranes and new bone were taken from the bone defect area of rats respectively. HE staining was used to observe the morphology of induced membrane， and the width of bone rabecular and the number of osteoblasts of new bone tissue were measured. Immunohistochemistry was used to detect the protein expression of BMP-2 and VEGF in induced membrane. Western blotting assay was used to detect the protein expression of Smad1， Smad4 and Smad7 in new bone. RESULTS： Compared with other 3 groups， the degradation of bone cement in drug-loading CPC group was more obvious in the bone defect areas， which showed that the formation of induced membrane was observed and the bone defect areas were smaller； capillary endothelial cells were abundant and orderly arranged in the induced membranes， and the width of bone trabeculae and the number of osteoblasts in the new bone tissue increased significantly （P＜0.05）； the protein expression of BMP-2 and VEGF in the induced membrane， the protein expression of Smad1， Smad4 and Smad7 in new bone were increased significantly （P＜0.05）. CONCLUSIONS： CPC loading total flavonoids of D. mariesii promotes the formation of induced membrane osteoblast in bone defect model rats， which may be associated with regulating osteoblast differentiation by activating BMP-2/Smad pathway； at the same time， it can promote bone healing by promoting the differentiation of vascular endothelial cells， accelerating the formation of capillary network and increasing the expression of vascular endothelial cells.