Rare diseases have been a major challenge for clinical medicine and public health challenge in China. One of the effective measures is to conduct proactive research on rare diseases to deal with the disease burden of the diseases. However, low prevalence, disperse distribution of patients, lack of knowledge about the disease course, and phenotype heterogeneity hamper the development of research for rare diseases. Recently, it has been found that patients registry is effective in understanding the course of the disease and accu- mulating the cases and data of clinical research or clinical trial design. At present, most of developed countries or regions in the world have promoted clinical research and clinical trials of new medications on rare diseases by using the registration of rare disease. In 2016, Peking Union Medical College Hospital established China's first registry system at the national level-National Rare Disease Registry System of China(NRDRS). NRDRS has accumulated 68 137 cases data registered by the researchers from China's 101 collaborating hospitals in 29 provinces/municipalities/autonomous regions, covering 171 different, and forming 188 cohorts. To date, NRDRS complete the initial stage of resources buildup.Nex stage will be focused on clinical research and clinical trials related to rare diseases based on NRDRS. This article is on the process of building NRDRS, the potential support for conducting clinical research and clinical trials related to rare diseases, and the challenges will be faced.

Folliculogenesis is essential for production of female gametes in vertebrates. However, the molecular mechanisms underlying follicle development, particularly apoptosis regulation in ovary, remain elusive. Here, we generated sox3 knockout zebrafish lines using CRISPR/Cas9. sox3 knockout led to follicle development retardation and a reduced fecundity in females. Comparative analysis of transcriptome between sox3 and wild-type ovaries revealed that Sox3 was involved in pathways of ovarian steroidogenesis and apoptosis. Knockout of sox3 promoted follicle apoptosis and obvious apoptosis signals were detected in somatic cells of stages III and IV follicles of sox3 ovaries. Moreover, Sox3 can bind to and activate the promoter of cyp19a1a. Up-regulation of Cyp19a1a expression promoted 17β-estradiol synthesis, which inhibited apoptosis in follicle development. Thus, Sox3 functions as a regulator of Cyp19a1a expression, via 17β-E2 linking apoptosis suppression, which is implicated in improving female fecundity.

Objective To investigate the expression of miRNA-140 in chondrocytes and synovial fluid of osteoarthritis (OA) patients, and explore the relationship between the miRNA-140 expression and OA severity.Methods This study enrolled 30 OA patients who underwent total knee arthroplasty for chondrocytes sampling and 30 OA patients who underwent intra-articular injection for synovial fluid sampling. All OA patients were grouped into mild [Kellgren and Lawrence (KL) grade 1-2], moderate (KL grade 3) and severe (KL grade 4), with 10 in each subgroups for each sampling purposes. 7 non-OA patients and 10 patients with knee injury were collected for cartilage and synovial fluid sampling respectively as control groups. Chondrocytes were isolated from the cartilage tissue and cultured in vitro. Quantitative real time PCR for miRNA-140 in chondrocytes and synovial fluid were performed, and the U6 snRNA was used as internal control. The expression difference of miRNA-140 among groups and correlation between the expression and the KL grade of OA were analysed using one-way ANOVA and Spearman test respectively. Results The expression of miRNA-140 in chondrocytes of knees in OA patients was reduced than that in normal knees, and the between-group difference was statistically significant (F=305.464, P<0.001). miRNA-140 could be detected in synovial fluid of both normal knees and OA knees, its relative expression level was reduced in synovial fluid of OA group compared with normal group, and the between-group difference was statistically significant as well (F=314.245, P<0.001). The relative expression level of miRNA-140 in both chondrocytes and synovial fluid were negatively correlated with the KL grade of OA(r=-0.969, P<0.001; r=-0.970, P<0.001). Conclusion miRNA-140 could be detected in chondrocytes and synovial fluid of OA patients, and its expression was negatively correlated with the severity of OA.
Adult ; Aged ; Cells, Cultured ; Chondrocytes ; metabolism ; Female ; Humans ; Knee Joint ; metabolism ; Male ; MicroRNAs ; analysis ; Middle Aged ; Osteoarthritis, Knee ; metabolism ; Real-Time Polymerase Chain Reaction ; Synovial Fluid ; metabolism

BACKGROUND:Oversize stress of a dental implant and its surrounding tissue is the main factor to affect the
long-term use of dental implants. So, the reasonable and precise design of implant shape is one of the important methods of prolonging the life span of dental implants.
OBJECTIVE:To make the optimal analysis and design of the diameters of connector screw and central screw of the adjustable-angle dental implant invented in the earlier stage.
METHODS: The finite element analysis model of the edentulous mandible with adjustable-angle dental implant was established by software Pro/E 5.0, Mimics 10.0 and ANSYS Workbench 14.5. The maximum equivalent
stress of dental implant-edentulous mandibular model was analyzed.
RESULTS AND CONCLUSION:The maximum equivalent stress of dental implant-edentulous mandibular model

Rhesus monkeys (Macaca mulatta) are human's closest evolutionary relatives next to Chimpanzees, and they are widely used in biomedical researches. Analyses of the rhesus monkey trasnscriptome and the sequence divergence between monkey and human are of importantce to the development of scientific analyses and to the application and interpretation of the results from animal experiments. In this study, we analyzed the genetic and transcriptional information. Four hundred and one clones were randomly selected from a liver tissue cDNA library of rhesus monkey, and the expressed sequence tags (ESTs) were sequenced. We acquired 393 effective ESTs that were assembled into 221 Unigenes with Phrap software. Alignments of the sequences showed that 188 Unigenes matched with known proteins in Swiss_prot database, of which 16 Unigenes matched the known rhesus proteins, and 171 Unigenes had high homology with human proteins. Then the result of BLASTN comparison showed that 26 of another 33 Unigenes matched the known rhesus genes. Finally, the remaining Unigenes were aligned in dbEST and rhesus genome database, and the results suggested 3 Unigenes be newly discovered ESTs of rhesus.
Animals ; Expressed Sequence Tags ; chemistry ; Gene Library ; Liver ; chemistry ; Macaca mulatta ; genetics ; Sequence Analysis, DNA

This study sought to clone Chinese Banna minipig inbred-line (BMI) alpha1,3-galactosyltransferase (alpha1,3-GT) gene and construct its recombinant eukaryotic expression vector. Total RNA was isolated from BMI liver. Full length cDNA of alpha1,3-GT gene was amplified by RT-PCR and cloned into pMD18-T vector to sequence. Subsequently, alpha1,3-GT gene was inserted into pEGFP-N1 to construct eukaryotic expression vector pEGFP-N1-GT. Then the reconstructed plasmid pEGFP-N1-GT was transiently transfected into human lung cancer cell line A549. The expression of alpha1,3-GT mRNA in transfected cells was detected by RT-PCR. FITC-BS-IB4 lectin was used in the direct immunofluorescence method, which was performed to observe the alpha-Gal synthesis function of BMI alpha1,3-GT in transfected cells. The results showed that full length of BMI alpha1,3-GT cDNA was 1116 bp. BMI alpha1,3-GT cDNA sequence was highly homogenous with those of mouse and bovine, and was exactly the same as the complete sequence of those of swine, pEGFP-N1-GT was confirmed by enzyme digestion and PCR. The expression of alpha1,3-GT mRNA was detected in A549 cells transfected by pEGFP-N1-GT. The expression of alpha-Gal was observed on the membrane of A549 cells transfected by pEGFP-N1-GT. Successful cloning of BMI alpha1,3-GT cDNA and construction of its eukaryotic expression vector have established a foundation for further research and application of BMI alpha1,3-GT in the fields of xenotransplantation and immunological therapy of cancer.
Animals ; Animals, Inbred Strains ; Base Sequence ; China ; Cloning, Molecular ; Galactosyltransferases ; genetics ; metabolism ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Recombinant Proteins ; genetics ; metabolism ; Sequence Analysis, DNA ; Swine ; Swine, Miniature ; genetics ; Transfection

In the conventional treatments of type I diabetes, there are various problems. As a new adequate treatment of diabetes, cell replacement therapy of diabetes has been applied and given research priority. We have investigated the applications of cell transplantation in the treatment of diabetes and have retrieved the relevant articles on cells transplantation for the treatment of diabetes. In this paper, we review the history, development, merits and demerits of cell transplantation and the recent advances in pancreatic islet transplantation research. The latest progress in the induction of stem cell to differentiate into the insulin-producing cells was also introduced.
Animals ; Diabetes Mellitus, Type 1 ; surgery ; therapy ; Humans ; Insulin-Secreting Cells ; cytology ; Islets of Langerhans Transplantation ; methods ; Stem Cell Transplantation

This study was aimed to create strontium-calcium sulfate compounds for making a new bioactive material with osteoconductive and osteoinduceable activity for bone repairing. Its mechanics and degradation features were assessed in vitro. Powders of alpha-calcium sulfate hemihydrate (alpha-CSH) and SrCl2 were mixed completely to make Sr-calcium sulfate compounds materials with 6 different concentrations (0%, 0.1%, 0.3%, 0.5%, 1% and 2%) of Sr. Scanning electron microscope was used to observe the configuration of the new materials. The compressive strength of each material was tested. The materials were soaked into simulated body fluid (SBF) to test the features of degradation, which included pH, weight loss, declination of compressive strength and the changes of strontium ion concentration. The crystal appearances were influenced by incorporating of strontium. The compressive strength of non-strontium incorporating calcium sulfate was 36.65 +/- 2.22 MPa. When the concentration of strontium was increasing, the compressive strength measurements of the materials tended to decline. The compressive strength declined to 20.56 +/- 2.64 MPa when the strontium concentration reached to 2%. The pH value of the SBF declined when the time of degradation increased, but both of them were very stable. All of the materials got weight loss after being soaked in SBF for several weeks. The weight loss was slight within 4 weeks and it became dramatic after 4 weeks. When the concentration of strontium was increasing, the weight loss became more rapid and significant (P<0.05). During 0-4 weeks' degradation in SBF, the materials' compressive strength decreased much slower when the strontium concentration was below 0.5%; however, when the decrement of strength became faster, the strontium concentration became higher. The concentration of strontium ion in SBF began to increase faster after 4 weeks' soaking in SBF. As the concentration of strontium was increasing, the strontium ion concentration in SBF became higher (P = 0.000). The new compound materials made by the mixing of alpha-calcium sulfate hemihydrate and SrCl2 can provide efficient compressive strength. The features of degradation of the materials are very stable. The new materials can release lots of bone inducible substance-strontium ions to repair bone defection after 4 weeks of degradation.
Bone Substitutes ; chemical synthesis ; chemistry ; Calcium Sulfate ; chemistry ; Compressive Strength ; Humans ; Osteogenesis ; Strontium ; chemistry

Mesenchymal stem cells from bone marrow of Rhesus monkey (RhBMSCs) were isolated by density gradient centrifugation, purified by adherence separation, and further identified by phenotype and karyotype analysis. Growth characteristics of RhBMSCs were investigated by observation of cell proliferation and detection of apoptosis. Density gradient centrifugation and adherence separation revealed a simple method to obtain fairly pure RhBMSCs. Fusiform was the most common form of cell under observation, and cell karyotype was normal. Phenotyping assay revealed that the RhBMSCs are highly positive (99.2%) for CD29 when compared with the low positive rates (less than 3.2%) for CD34, CD45 and HLA-DR, which indicated a successful isolation of high purity RhBMSCs and a vigorous activity of cells to proliferate. But the proliferation activity of RhBMSCs gradually decreased following the increasing of cell generations. The methods and results of isolation, expansion, identification and growth characteristics of RhBMSCs were discussed in this paper, which may be helpful to understanding the bone marrow mesenchymal stem cells of human, and may serve as the groundwork for orientated differentiation of RhBMSCs and tissue repairment on experimental animal model of rhesus monkey.
Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; Cell Separation ; Cells, Cultured ; Integrin beta1 ; blood ; Macaca mulatta ; Mesenchymal Stromal Cells ; cytology ; Phenotype

To evaluate the efficiency of three in vitro refolding methods for a humanized single-chain Fv antibody against human CTLA4(CD152) expressed in E. coli, the denatured and purified inclusion bodies (IBS) were refolded by dilution, dialysis and in situ refolding via Immobilized Metal-Ion-Affinity Chromatography (IMAC), respectively. The concentration of refolded scFvs was examined by Bradford method. And the antigen binding activity of the refolded scFvs was analyzed by indirect cell-ELISA. The highest and lowest refolding yields could be obtained by dialysis and in situ refolding via IMAC, respectively. The binding activity of the refolded scFv by dialysis was 1.95-fold higher than that by dilution, 4.13-fold higher than that by in situ refolding via IMAC (GSH/GSSH excluded) and 3.63-fold higher than that by in situ refolding via IMAC (GSH/GSSH included), respectively. In conclusion, a high refolding yield and binding activity of scFv with natural conformation could be obtained by dialysis in the condition of 0. 15 mol/L sodium chloride, 50 mmol/L Tirs-HCl, pH 8. 0 buffer containing 3 mmol/L reduced glutathione and 1 mmol/L oxidized glutathione for 48 hours at 4 degrees C.
Antigens, CD ; biosynthesis ; genetics ; immunology ; Antigens, Differentiation ; biosynthesis ; genetics ; immunology ; CTLA-4 Antigen ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; immunology ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology