Objective To explore the effect of fluoride and arsenic pollution on bone metabolism in exposed population. Methods One hundred and fifty-two fluoride and arsenic exposed people were selected from Jiaole village, Yuzhang town, Xingron county, Guizhou province in 2006, and 59 not exposed people from Daguoduo village 13 km away from Jiaole village were selected as control. Urinary fluorine(UF), urinary arsenic (UAs), urinary hydroxyproline (UHYP), cross-linked N-telopeptides of type I collagen (UNTX) and bone strength index(STI) were detected. Results The main effect of fluoride on UHYP and UNTX were statistically significant (F = 9.785, 4.225, P < 0.01 ), but was not significant on STI(F = 0.183, P > 0.05). The main effect of arsenic on UNTX was statistically significant (F = 2.660, P < 0.05 ), but was not significant on UHYP and STI(F = 2.012, 0.183,all P > 0.05). The interaction between fluoride and arsenic on UNTX was statistically significant (F= 2.429, P <0.01), but was not significant on UHYP and STI(F= 1.218, 1.001, all P> 0.05). Conclusions Fluoride exposure can affect the metabolism of collagen and bone resorption, and Arsenic exposure main affect bone resorption, fluoride and arsenic co-exposure have more significant effect on bone resorption. UNTX may be used as biological biomarker of bone metabolism for population co-exposed to fluoride and arsenic in health monitoring.

AIM:To be one of the primary cause injury to multiple sites of ocular of glaucoma which affects over 70 million people worldwide. We applied data mining techniques, linear and the matrix operations, efficiently calculated the network and estimated the possible function of the“node” genes of the retina and optic of glaucoma, in order to provide new thought and method on the pathogenesis of glaucoma.
METHODS: The data in this study is from Gene Expression Omnibus ( GEO ) which belong to Nation Center for Biotechnology Information ( NCBI) , the quality of the raw data CEL files was processed and analyzed by the Expression software which belong to Affymetrix Inc. , Santa Clare, CA, USA. Significant analysis method ( SAM) which base on the T test was used to identified the significant genes. Based on GRNInfer and Gvedit soft we set up gene networks of optic and retina of mice and further more enriched analysis which based on DAVID and MAS3. 0 online software were processed.
RESULTS:The analysis between the group of the optic nerve heads and retinas in different stage of glaucoma showed that the amount of significant different expressed genes in the optic never head group increased significantly comparing with the group of retina in the early stage of glaucoma, the analysis of the genes network construction show that:the node genes of optic nerve heads included Unc13c、Kif5a、TRPM1、PANX; and the node genes of retina include POU4F1, NEFL, BC03870, CALB2. Metabolic pathways enrichment analysis which based on MAS3. 0 online platform show that there was mainly the amyotrophic lateral sclerosis, tyrosine metabolism, melanogenesis, Nitrogen metabolism, Gap junction, Leukocyte transendothelial migration metabolism pathway enriched out in optic nerve head; and there was mainly amyotrophic lateral sclerosis, neurodegenerative disorders, prostate cancer, leukocyte transendothelial migration metabolism pathway enriched out in retina.
CONCLUSION:By understanding bioinformatics result, it seems optic were more sensitive than the retina to high intraocular pressure, and weather high expression of TYrp1 gene can be as a sensitive diagnostic item require more evidence back up. Functional enrich analysis of node gene showed that cytoskeleton reconstructed, molecular motor and nutrients transport function improve in optic; and in retina, the most prominent finding in retina was enrichment function modules were focus on regeneration, repairing and differentiation of cells, which remind that we should reinforce research on reparation of retina of primary glaucoma. Metabolic pathways enrichment analysis show that inflammatory response plays prominent place in optic and retina of primary glaucoma, because of the optic narrow and crowed anatomic shape, nutrient metabolism and substances transfer enrichment modules play an important role in optics of primary glaucoma.

As a depot drug delivery system, injectable polylactide-polyglycolide (PLGA) sustained-release microspheres have been successfully used to treat many diseases since the first microsphere product Lupron depot was approved for marketing in the United States in 1989. It has the ability of long-term release in the body for several days to several months, which can not only reduce the times of administration, but also reduce the drug blood concentration fluctuations, significantly improve the safety and patient compliance. In vitro-in vivo correlation (IVIVC) makes the development of microspheres more possible. It can describe the dynamic information of drug release in vivo through the in vitro release behavior of microspheres, and can reduce the workload of each stage and shorten the time span while characterizing the performance of microspheres. IVIVC can provide guidance or support for drug development, production changes, supervision and management. This article summarizes the release mechanism of injectable PLGA sustained-release microspheres, common measurement methods and theories of in vitro and in vivo release. And we also focus on the establishment and application of IVIVC, especially A level IVIVC in the field of microsphere preparations, to provide a reference for further study on in vitro-in vivo correlation of microspheres.

The cDNA encoding human Vascular Endothelial Growth Factor 165 (VEGF165) was amplified using RT-PCR from human tonsil tissue and cloned into eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid pcDNA/V was transferred into 293 cells mediated by liposome and the cells stably expressing VEGF were selected under the pressure of G418. ELISA and Western blotting demonstrated that the eukaryotic expression vector pcDNA/V was successfully constructed and its corresponding protein could be expressed efficiently in vitro. Chick Charioallantoic Membrane (CAM) bioassay showed that recombinant protein has biological activity of hVEGF. Model rats with acute myocardial ischemia were used to further study the expression of VEGFin vivo. The model rats were divided randomly into three groups: control group, pcDNA3.1 (+) group and pcDNA/V group. 50microL naked plasmid DNA or saline was intramyocardially injected at three sites into the border zone of infarction. The hearts of rats were excised and fixed histologically, then the infarction sizes were studied by immunohistochemical staining and electron microscope after four weeks. Immunohistochemical staining for VEGF appeared to be negative in control and pcDNA3.1 (+) groups. In pcDNA/V group, myocardial cells in infarction border zone showed positive staining for VEGF in cytoplasm. Ultrastructural anaylsis showed that there were visible hyperplasia of vascular endothilium in pcDNA/V group. The control and pcDNA3.1 (+) groups showed less capillary hyperplasia. In this study, VEGF165 gene was successfully cloned and its protein expressed in vitro and in vivo was of bioactivity, which provides a basis for the further study of biological functions of human VEGF.
Animals ; Cell Line ; Chickens ; Chorioallantoic Membrane ; blood supply ; Disease Models, Animal ; Genetic Therapy ; Humans ; Male ; Myocardial Infarction ; metabolism ; pathology ; therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

Actins ; metabolism ; Angiomyolipoma ; metabolism ; pathology ; surgery ; Desmin ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Perivascular Epithelioid Cell Neoplasms ; metabolism ; pathology ; S100 Proteins ; metabolism ; Skin Neoplasms ; metabolism ; pathology ; surgery

According to the principle of evidence-based medicine (EBM), Chinese medical literatures based descriptive statistical analysis of common Chinese medical syndrome types were performed. By data extraction, standardization, and frequency calculation of disease names and syndrome types from 286 literatures in line with the inclusion criteria, the frequencies of diseases and syndromes were obtained to analyze common syndrome types in clinical practice, to analyze the distribution features of disease related syndromes and syndrome related diseases, to analyze the distribution of basic Chinese medical syndrome types in clinical common diseases as a whole, thus providing reference for clinical and basic researches.
Evidence-Based Medicine ; Humans ; Medicine, Chinese Traditional

Objective To knockout Rag2 and IL2rg genes and construct severe combined immunodeficiency mice based on CRISPR/Cas9 technology. Method Design and synthesis of 25 bp sgRNA were made according to the Rag2 and IL2rg sequences in Genbank. After annealing, sgRNA was cloned into pX330 vector. Recombination plasmid Rag2?sgRNA, IL2rg?sgRN and Cas9 were then transcribed into RNA, these RNA were microinjected into zygotes and the zygotes were transplanted into recipient ICR mice. F0 founders were born and mutated F0 founders mated with wild type mice to obtain F1 generation heterozygous mice. Mutated F1 mice were crossed and got F2 generation homozygous mice. Genotype and phenotype of the knockout mice were identified by sequencing, flow cytometry and xenograft model. Results Rag2?sgRNA and IL2rg?sgRNA recombination plasmids were constructed and transcribed into RNA. After microinjection and mat? ing, F0 founders were born and F2 homozygous mice were obtained. The results of sequencing showed that there were two types of genotype in IL2rg gene, 10 bp or 11 bp deletion;however, there was only one genotype in Rag2 gene, which was 8 bp deletion. Compared with wild?type BALB/c mice, the number of CD3 +, B220 + and NKp46 + cells in peripheral blood of the knockout mice was reduced significantly. After inoculation of human breast cancer cell line SKBR?2HL cells, tumor size in the xenograft mouse model was increased gradually along with time extension. Conclusion CRISPR/Cas9 is an efficient way to mutate Rag2 and IL2rg gene in mice in vivo, leading to aberrant T cells, B cells and NK cells.

Objective To construct miRNA-29b1 gene knockout mice based on CRISPR/Cas9 technology. Methods To design and synthesize sgRNA according to the miRNA-29b1 sequence in Genbank .sgRNA and Cas9 were transcribed to RNA in vitro, these RNA were then microinjected into zygotes of C 57BL/6 mice.After mouse birth, the genome DNA was extracted and sequenced to identify its genotype; meanwhile , real-time PCR was used to assay the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney of mutated mice .Result A 20 bp sgRNA targeted on miRNA-29b1 was synthesized and transcribed to RNA with Cas 9.After microinjection, miRNA-29b1 gene-mutated mice were obtained.The sequencing results showed that there were two types of genotype for the mutated mice , one was 10 bp deletion, and another was 23 bp deletion accompanied with a 3 bp insertion.Compared with the wild-type mice, the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney was reduced significantly .Conclusions miRNA-29b1 gene knockout mice are constructed successfully by using CRISPR /Cas9 technology.

OBJECTIVETo observe the change of PTEN/PI3K/AKT pathway hypoxia-inducible factor (HIF-1α), vascular endothelial growth factor (VEGF) in rats with adjuvant arthritis and to explore the mechanism of neovasculization in rheumatoid arthritis.

METHODSThirty rats were randomly divided into normal control group and model control group. The model control group were established the model of adjuvant arthritis using Freund's complete adjuvant. At 19 days after modeling, the expression of microvascular density (MVD), HIF-1α, VEGF were detected by ELISA assay and PTEN, PI3K, AKT were detected by Werstern Blotting.

RESULTSCompared with the normal control group, paw swelling, arthritic index were increased, and the expression of MVD, VEGF, HIF-1α of serum, PI3K, AKT of synovial tissue were significantly increased, PTEN was significantly decreased in model control group. PI3K, HIF-1α were positively correlated with MVD; VEGF, AKT were positively correlated with paw swelling; PTEN was negatively correlated with the arthritis index; HIF-1α was positively correlated with VEGF; PI3K was positively correlated with AKT, PTEN was negatively correlated with PI3K, AKT, VEGF.

CONCLUSIONImbalance of PTEN/PI3K/AKT pathway in rats with adjuvant arthritis is one of the mechanisms of synovial neovasculization.

Animals ; Arthritis, Experimental ; physiopathology ; Hypoxia-Inducible Factor 1, alpha Subunit ; physiology ; Neovascularization, Pathologic ; etiology ; PTEN Phosphohydrolase ; physiology ; Phosphatidylinositol 3-Kinases ; physiology ; Proto-Oncogene Proteins c-akt ; physiology ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; physiology

OBJECTIVETo evaluate the correlation between blood eosinophil (EOS)level and steroid doses in patients of bullous pemphigoid (BP).

METHODSA total of 82 untreated BP inpatients (n=49) and outpatients (n=33) were enrolled in this study. The blood EOS level and the steroid doses before and after treatment were recorded. The correlation between EOS level and steroid doses was analyzed retrospectively.

RESULTSEOS increased in 69 BP patients (84.15%); on the contrary, only 10% of normal controls had increased EOS (t=1.99,P<0.001). In 44 inpatients, the blood EOS remained high before steroid treatment, and quickly returned to normal level after the disease became stable. There was a linear correlation between EOS and steroid doses (Spearman analysis,r=0.496,P<0.001). In 5 patients who were treated by non-steroid approach, EOS level also declined after the disease was resolved.

CONCLUSIONEOS can be one of useful indicators for the application of steroids in the treatment of BP.

Adult ; Aged ; Aged, 80 and over ; Eosinophils ; Female ; Humans ; Leukocyte Count ; Male ; Middle Aged ; Pemphigoid, Bullous ; drug therapy ; immunology ; Retrospective Studies ; Steroids ; administration & dosage ; therapeutic use