1.Detection and analysis of the Rhodopsin gene in a consanguineous Chinese Han autosomal dominant retinitis pigmentosa family
Xue-juan, CHEN ; Xiang, GAO ; Chen, ZHAO ; Kan-xing, ZHAO
Chinese Journal of Experimental Ophthalmology 2013;(6):602-607
Background Rhodopsin (RHO) gene is the most common disease gene for autosomal dominant retinitis pigmentosa (adRP),one of the main pathogenesis is that misfolded mutant RHO proteins accumulate in the endoplasmic reticulum and cause endoplasmic reticulum stress (ERS).Objective This study aimed to determine the genetic basis for a consanguineous Chinese Han adRP family.Methods This study procedure complied with Helsinki Declaration.All participants in the family were investigated under the informed consent.Regular ocular examination was performed on the patients in this family.Next-generation sequencing (NGS) was carried out to screen the mutations in 189 genes associated with hereditary retinal diseases (HRDs).After being analyzed and filtered,variations detected by NGS were validated by Sanger sequencing and evaluating of pathogenicity.The wild-type RHOWT and mutant RHOP53Rwere cloned into the vector pEGFP-N1.Then the two plasmids were transfected into adult retinal pigmentosa epithelium cell line(ARPE19) and human embryo kidney 293 line (HEK293) to observe the location of rhodopsin-GFP fusion protein in cells,and the expression of ERS related protein XBP1 in the cells was detected by quantitative-PCR and Western blot.Results This family included 5 generations with the typical adRP characteristics.Genetic analysis identified a heterozygous variation,p.P53R in RHO gene,which was fully cosegregated in the family.Wild-type RHOWT-GFP fusion proteins showed the green fluorescence on the endoplasmic reticulum and cytomembrane,but the misfolded mutant RHO-GFP fusion protein gathered only in endoplasmic reticulum.Compared to wild-type RHOWT,the XBP1 was activated and increased by (1.28 ±0.09) fold.The introns of 26 bases in XBP1 mRNA were removed in the HEK293 cells with mutant RHO-GFP fusion protein,and the expression of XBP1 was stronger in the HEK293 cells with mutant RHO-GFP than that in HEK293 cells with wild type RHO-GFP and cells with blank pEGFP-N1 plasmid.Conclusions Heterozygous variant RHO p.P53R is very likely the pathogenical mutation in the adRP family.The RHOP53R mutant rhodopsin protein can not be delivery effectively from the endoplasmic reticulum to the cell membrane,and these proteins accumulate in the endoplasmic reticulum,which causes ERS.
2.The value of diagnosing kidney neoplasms with 18F-FDG PET/CT
Jianhua SONG ; Jinhua ZHAO ; Xiang CHEN ; Yan XING
Chinese Journal of Nuclear Medicine and Molecular Imaging 2013;(3):184-187
Objective To explore the clinical diagnostic value of 18 F-FDG PET/CT in patients with kidney neoplasms.Methods Seventy-nine patients (52 males,27 females,average age (57.3 ± 14.1) years),who had definitive diagnosis of kidney neoplasms by 18F-FDG PET/CT and pathological or clinical comprehensive diagnosis in recent five years,were analyzed retrospectively.The diagnosis by PET/CT was made according to the changes of kidney shape,tumor density and FDG uptake (SUVmax).The diagnostic efficacy was calculated.Results Among the 79 patients,70 cases were finally diagnosed as malignant tumors (including 40 cases of renal cell carcinoma,5 cases of renal pelvis carcinoma,8 cases of lymphoma,16 cases of metastatic tumor,and 1 case of renal fascia capsule liposarcoma) and 9 cases of benign tumors (including 7 cases of angiomyolipomas,1 case of renal acidophilic cell adenoma,1 case of metanephric adenoma; the benign tumors did not contain small lipid sample hamartoma cases).The detective rate of kidney neoplasms with 18F-FDG PET/CT was 97.5% (77/79).For the identification of benign and malignant lesions,the sensitivity of 18F-FDG PET/CT was 92.9% (65/70),specificity was 7/9,accuracy was 91.1% (72/79),positive predictive value was 97.0% (65/67),and negative predictive value was 58.3 % (7/12).Conclusions 18 F-FDG PET/CT can detect and identify most of kidney tumors.Whole-body checking and comprehensive evaluation on kidney cancer patients are still the main advantages of PET/CT.
3.Isolation proliferation and characterization of endothelial progenitor cells from adult peripheral blood.
Xing-xiang WANG ; Jun-hui ZHU ; Jun-zhu CHEN
Chinese Journal of Applied Physiology 2005;21(1):94-95
Adult
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Cell Separation
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methods
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Endothelial Cells
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cytology
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Humans
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Stem Cells
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cytology
4.Neural stem cells on a novel composite scaffold:growth and differentiation
Ran XING ; Xuyi CHEN ; Xiang ZHU ; Ruixin LI ; Yue TU
Chinese Journal of Tissue Engineering Research 2016;20(19):2857-2863
BACKGROUND:Neural stem cel s with self-proliferation and differentiation potential are the ideal seed cel s for central nervous tissue engineering. Although col agen and silk fibroin as biological scaffold materials have been widely used, both of them used alone have certain shortcomings. Is it possible to combine the two materials to build a novel neural tissue-engineered scaffold? What is the effect of this novel scaffold on the growth and differentiation of neural stem cel s? OBJECTIVE:To observe the growth and differentiation of neural stem cel s seeded onto the novel composite scaffold. METHODS:The rat embryonic neural stem cells were inoculated onto new composite scaffolds, and then, their growth and differentiation were observed by light microscopy and scanning electron microscopy. Neural stem cells were cultured in conventional suspension culture as control group. Cell counting kit-8 assay was used to detect viability of neural stem cells in the two groups. Three-dimensional composite scaffolds carrying neural stem cells were slic ed into paraffin sections to observe the growth and differentiation of neural stem cells by hematoxylin-eosin staining and immunofluorescence staining. RESULTS AND CONCLUSION:Neural stem cel s cultured on the new composite scaffold grew and differentiated wel , and interconnected synapses were observed. Cel counting kit-8 assay showed that neural stem cel s on the scaffold grew wel , and the cel viability was significantly higher in the composite scaffold group than that in the control group (P<0.05). Hematoxylin-eosin staining and immunofluorescence staining of paraffin sections further provided evidence for good growth and differentiation of neural stem cel s on the scaffold. These results indicate that the novel composite scaffold with good biocompatibility benefits the growth and differentiation of neural stem cel s, promising a favorable application prospect.
5.Recent Research Situation of Heart Rate Variability-based Analysis of the Regulating Effect of Acupuncture on Autonomic Nervous Function
Zhifeng CHEN ; Yunyun ZHAO ; Xing WANG ; Yunxia XIANG ; Ling ZHAO
Shanghai Journal of Acupuncture and Moxibustion 2016;35(6):754-757
Traditional Chinese medicine considers that the final purpose of acupuncture treatment for disease is to harmonize yin and yang. Acupuncture redresses a balance between yin and yang by transmitting information to acupoints and then to corresponding visceral tissues and organs and the brain through the meridian system or the nervous system. In recent years, scholars at home thoroughly studied heart rate variability (HRV) analysis for acupuncture treatment of diseases and found that the regulating effect of acupuncture on visceral organs was closely related to autonomic nerves and acupuncture could restore a balance between sympathetic and parasympathetic nerves. Many scholars explored the effect of acupuncture on diseases related to autonomic nervous dysfunction by HRV frequency domain or time domain analysis and gained a certain achievements. This article summarizes recent years’ domestic studies on HRV-based analysis of acupuncture for diseases with autonomic nervous dysfunction and reviews the close relationship between autonomic nerves and diseases and the related mechanism of acupuncture action.
6.Application of collagen and silk fibroin scaffolds in spinal cord tissue engineering
Xiang ZHU ; Xuyi CHEN ; Yingfu LIU ; Ran XING ; Yue TU
Chinese Journal of Tissue Engineering Research 2014;(39):6359-6363
BACKGROUND:Col agen and silk fibroin materials for construction of spinal cord scaffolds have been proven to repair or partial y repair damaged spinal cord nerve function. OBJECTIVE:To introduce partial characteristics of the col agen and silk fibroin and to review the recent progress and application as scaffolds in spinal cord tissue engineering. METHODS:A computer-based search of CNKI and PubMed databases (2003-01/2012-10) was performed for articles addressing the application of col agen and silk fibroin scaffolds in spinal cord injury with the keywords of“col agen, silk fibroin, scaffold, spinal cord injury”in Chinese and English, respectively. RESULTS AND CONCLUSION:Col agen has low antigenicity, good biocompatibility and biodegradability. Col agen and its degradation products can cause no inflammatory reactions in the body, but have the disadvantages of rapid degradation and poor mechanical properties. Silk fibroin has good biocompatibility and excellent mechanical properties, but its degradation is slow. The col agen and silk fibroin are compounded using an electrostatic spinning technology to improve the physical properties of the material on the basis of maintaining good biocompatibility. At present, fibroin or col agen materials in terms of nervous system repair have been studied, laying some foundation for spinal cord tissue engineering. Considering the similar characteristics and mechanics performance to the spinal cord tissue, col agen/silk fibroin composite materials are expected to become the ideal scaffold materials for spinal cord tissue engineering.
7.Cycloplegic effects of 0 . 5 g/L tropicamide in various low myopia age groups
Hai-Rong, ZHANG ; Hong-Xing, DIAO ; Xiang, CHEN
International Eye Science 2014;(12):2295-2296
AlM: To investigated the refraction and cycloplegic effects of 0. 5g/L tropicamide eye drops in various low myopia age groups, and provide clinical guidelines for proper usage of cyclopegic agent in different population.METHODS:Three low myopia age groups (n=45) were selected, non-cycloplegic and cycloplegic auto-refraction using “Shin-Nippon” auto-refractor were performed at various time point for refractive error. The subjective accommodation amplitudes were also collected at each time point.
RESULTS: The accommodation amplitude showed a similar but significant decline between 20~60min after the first cyclopegic drops in all three age groups. However, the decline of accommodation amplitudes were not statistically correlated with the results of “Shin-Nippon”auto-refraction results at different time points.
CONCLUSlON: The refraction time should be arranged according to the cycloplegic effects of tropicamide. Open-field auto-refractor could reduce the usage of cycloglegice drugs in low myopia groups, even for the youngest.
8.High-sensitive Detection Method for Siderophores from Pseudomonas
Shao-Xing CHEN ; Xiang ZHAO ; Ping SHEN ; Zhi-Xiong XIE ;
Microbiology 1992;0(03):-
CAS (Chrome azurol S) assay was a universal method for detecting the bacterial siderophores, and Sugar-Asn liquid medium has been applied to the studies on siderophores from Pieudomonas. In this paper, Asp has been substituted for Asn, and MSA-CAS agar plate was developed by integrating the MSA (sugar-Asp) medium and CAS bright blue dye, which has been used in the universal CAS assay. On the aspect of siderophores detection , 8 strains of 7 species from Pseudomonas had been screened on MSA-CAS agar plates and universal CAS assay respectively. The results showed that MSA-CAS agar was higher-sensitive and lower basic fluorescent than universal CAS assay.
9.Improvement and Analysis in Over- siderophores Production Bacteria Filtrating and Detecting
Xiang ZHAO ; Zhi-Xiong XIE ; Shao-Xing CHEN ; Ping SHEN ;
Microbiology 1992;0(06):-
A novel stable blue agar plate which is convenient to preparing and more effective than the universal chrome azurol sulfonate (CAS) assay established by Schwyn and Neilands was designed by replacing MM9 growth medium and pipes with certain concentrate of phosphorus buffer solution which pH could be stabled at 6.8. It is more suitable for screening over- siderophores production bacteria. Since OD_ 630 of the sample is usually out of the range of spectrophotometer with CAS assay solution when quantifying the siderophores and the outcome is not steady,the measuring wavelength had been changed to 680 nm corresponding to the middle of max absorbance and the correlation between siderophores concentrations and OD was unchanged. But the detecting sensitivity is elevated by enlarged the absorbance differences among samples with different productivity of siderophores at 680 nm .
10.Association study between schizophrenia and polymorphism of phosphoserine aminotransferase 1gene
Juan GUO ; Yuantang CHEN ; Changjiang HE ; Li ZHANG ; Yu WU ; Yangling XING ; Lei AO ; Xiang CHEN
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(11):1019-1021
ObjectiveTo detect the association between schizophrenia and polymorphism of phosphoserine aminotransferase 1 ( PSAT1 ) gene.MethodsThe study group included 158 patients with schizophrenia from Xi' an Mental Health Center and the control group included 316 parents.The polymorphism of rs69287125,rs137824326 of phosphoserine aminotransferase 1 gene was detected with PCR methods and SNP typing in all nucleus families by correlation analysis and haplotype relative risk analysis.ResultsThe rs69287125 allele was associated with schizophrenia (P=0.011 ),the G allele was protective factor (Z =-2.31 ) and the A allele was hazarding factor (Z =2.31 ).The rs137824326 allele was associated with schizophrenia (P=0.007 ),the G allele was protective factor ( Z =- 2.54) and the A allele was the hazarding factor( Z =2.54).The haplotypes of A/A and G/G in the rs69287125-rs137824326 were associated with schizophrenia (P =0.021,0.015,Z =2.16,- 1.85).ConclusionThe polymorphism of phosphoserine aminotransferase 1 gene is associated with schizophrenia in Chinese.