1.Current research progress of choroidal metastasis
Lin-Hui, YUAN ; Xiao-Long, CHEN
International Eye Science 2016;16(7):1216-1220
Choroidal metastasis is one of the most common malignant tumors inside the eyes. It causes pain, hypopsia and some other related symptoms. It reduces the quality of the patients' life. It's significant for the patients to be detected and treated early, therefore they will have better vision and longer life. The treatments of choroidal metastasis are developing quickly. Both the vitreous cavity injection of targeted drug and gene therapy are hot topics of research. This paper summarizes the etiology, development, diagnosis and treatment of choroidal metastasis nowadays.
2.Preparation of recombinant firefly luciferase by a simple and rapid expression and purification method and its application in bacterial detection
Qin XIAO ; Hui CHEN ; Jinming LIN
Journal of Pharmaceutical Analysis 2010;22(2):97-101
A simple and rapid expression and purification method of recombinant firefly luciferase was developed for bacteria detection. A modified luciferase gene from North American firefly Photinus pyralis was cloned into pET28a expression vector and the recombinant protein was produced in Escherichia coli BL21. The recombinant luciferase, equipped with a polyhistidine affinity tag, was purified by immobilized metal ion affinity chromatography (IMAC). The approach generated an abundant expression and an efficient purification of a recombinant luciferase with final yield 1.995mg/L of cell culture. Experiments on the recombinant luciferase also showed that the relative light units (RUL) of the enzyme were 5.8×108, and the specific activity was 2.9×1010 RLU/mg. By applying adenosine triphosphate (ATP) bioluminescence to detection of the coin bacteria using the recombinant protein, the ATP content of bacteria was 9.48×10-16mol/mL, and was identical to the bacteria counts (4500CFU/mL) in order of magnitude. Taken together, our results provided a simple and efficacious method of the preparation of recombinant luciferase, which could be applied in the determination of bacteria via ATP bioluminescence.
3.Changes in gene-expression profiling in kidney of rats with sepsis
Xiongjian XIAO ; Jiandong LIN ; Yi CAI ; Beilei ZHANG ; Hui LIN
Chinese Journal of Emergency Medicine 2010;19(9):909-915
Objective To investigate the gene-expression profile in kidney of rats during late sepsis (24hours) by using microarray technology in order to offer some clue to revealing the pathogenetic mechanism of sepsis at gene level. Method A total of 30 Wistar rats were selected and divided into model group and control group randomly(random number). The rats of control group were sham operated and the rats of model group received cecal ligature and puncture (CLP) operation. The biomarkers of renal function were assayed and the histopathological changes of kidney in rats were observed under transmission electron microscope 24 hours after operation. Gene chips containing 22 107 rat-genes cDNA were used to exmine gene-expression in kidney of septic rats to sieve the genes with different expressions with software. Data were analyzed by using SPSS version 11.0 software package.Statistical analyses of two independent samples carried out by using t -test. Results Compared with the control group, the levels of blood urea nitrogen (BUN) and creatinine (Cr) of sepsis group were higher (P < 0.01 ). The histopathological changes in kidney of rats demonstrated the establishment of sepsis model successful 24 hours later.Compared with the control group, there were 325 genes with differential expression in model group. Among the known-functional genes, there were 100 up-regulated and 64 down-regulated. Sorted by biological function, the genes were mainly related to metabolism, immunoresponse, cellular signal transduction, apoptosis, ion channel,growth factor and so on. Conclusions A sequence of genes expressed differentially in kidney of rats with late sepsis. Microarray technology played an important role in the research into sepsis mechanisms.
4.Churg-Strauss syndrome in a child.
Yuan-yuan XIAO ; Lin MA ; Hui-min LI
Chinese Journal of Pediatrics 2010;48(7):545-546
Child
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Churg-Strauss Syndrome
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pathology
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Female
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Humans
6.Effects of chronic hypoxia on left and right ventricular function and the expression of cardiac TRPC channels in rats.
Hui-Qin CHEN ; Mo-Jun LIN ; Xiao-Ru LIU
Chinese Journal of Applied Physiology 2014;30(3):274-278
OBJECTIVETo explore the effects of chronic hypoxia on left and right ventricular function and the expression of cardiac transient receptor potential canonical (TRPC) channels in rats.
METHODSForty eight SD male rats were randomly divided into control group (CON) and chronic hypoxic pulmonary hypertension model group (CH) (n = 24). In CH group, rats were exposed in chronic hypoxia environment (10% +/- 0.2% O2) to induce myocardial hypertrophy. After 3 weeks, mean systemic arterial pressure (mSAP), right ventricular systolic pressure (RVSP), left ventricular systolic pressure (LVSP), left or right ventricular pressure maximum rate of rise (LV/RV + dp/dt(max)), left or right ventricular pressure maximum rate of descent (LV/RV-dp/dt(max)), right ventricular hypertrophy index (RVMI) and left ventricular hypertrophy index (LVMI) were measured. Left and right ventricular myocardium tissue sections were stained by HE and observed under light microscope. Real-time polymerase chain reaction (real-time-PCR) and Western blot were performed to detect the expression of TRPC subfamily.
RESULTSRVSP, RVMI, RV + dp/dt(max) and RV-dp/dt(max) were markedly elevated in CH group (P < 0.01) in comparison to CON group. LVMI was markedly reduced in CH group in comparison to CON group (P < 0.01). LVSP, LV + dp/dt(max) and LV- dp/dt(max) had no significant changes in CH group in comparison to CON group. Right ventricular myocardial cells of CH group became thick, the nuclei stained deeply, the shape of nuclei became not regularity. Left ventricular myocardial fibers did not change significantly. There was significant difference in the levels of mRNA and protein of TRPC1 between CON and CH groups.
CONCLUSIONFor three weeks exposed to chronic hypoxia induced right ventricular hypertrophy specifically, raised the mRNA and protein expression of TRPC1 on right ventricular myocardial cells . TRPC1 might be involved in the development of cardiac hypertrophy.
Animals ; Disease Models, Animal ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Hypoxia ; metabolism ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley ; Transient Receptor Potential Channels ; metabolism ; Ventricular Function, Left ; physiology ; Ventricular Function, Right ; physiology
9.Prokaryotic expression and application of human herpes virus 8 small capsid protein encoded by open reading frame 65
Wei-Lin YU ; Hui LI ; Xiao-Feng WANG ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To construct prokaryotic vector and express human herpes virus 8 (HHV8) small capsid protein open reading frame 65 (ORF65) in Escherichia coli.Methods DNA was extracted from BCB-1 cells (HHV8-positive but EBV-negative).HHV80RF65 coding sequence was amplified by PCR and inserted into the prokaryotic expression vector pThioHisA.Recombinant plasmid was transformed into E.coli BL21 to express fusion protein induced by IPTG.The expressed products were purified by affinity chromatography on Ni-NTA resin.The antigenieity and the specificity of the recombinant proteins was identified by Western blot with HHVS-positive serum samples.ELISA coating with the recombinant proteins was used to screen 568 serum samples,which were simultaneously detected by IFA kit(Biotrin) and ELISA kit(Qiagen).Results Gene sequencing showed that the target gene was identical with that of HHV8 standard species,the 31 500 fusion protein was found in SDS-PAGE.ELISA coating with the recombinant proteins had a good agreement with IFA kit(Biotrin) and ELISA kit(Qiagen).The detection for the clinical samples showed the ELISA kit was feasible,applicable and consistant.Conclusion The recombinant proteins showed good antigenicity,and it is valuable for further study on HHV8 specific antibody detection.