1.Cloning,expression and the immune characterization of panallergen profilin in coco(Cocos nucifera Linnaeus)
Guang MENG ; Min YAO ; Zhigang LIU ; Yulan WU ; Hongyun ZHANG
Chinese Journal of Immunology 1985;0(06):-
Objective:To clone and express panallergen profilin from the pollen of coco(Cocos nucifera Linnaeus).Methods:RT-PCR and RACE methods were applied to clone the full-length panallergen genes from coco pollen and the sequence was analyzed.The specific primers were designed.The ORF of profilin of coco pollen was amplified with RT-PCR and cloned into the expression vector pET 28a.Expression of the recombinant coco pollen profilin was carried out in E.coli BL21(DE3) and the purification of the recombinant protein was performed via affinity chromatography with Ni2+ coupled to sepharose.IgE reactivity to recombinant coco pollen profilin was investigated by immunoblot.Results:The complete sequence of coco pollen profilin was cloned.The sequence was 608 bp and included an open reading frame(396 bp) coding for 131 amino acids.Sequence analysis showed that the deduced protein was an acidic protein with an estimated molecular mass of 14.19 kD and a pI of 4.61.The GeneBank accession number of the clones was EF173598.After overexpressed in E.coli BL21(DE3),the recombinant protein was purified through affinity chromatography with Ni2+ coupled to sepharose.Immunoassay showed that the recombinant allergen has good IgE binding capacity.Conclusion:The profilin of coco pollen is expressed successfully in BL21(DE3),which will be used as a base for further study on coco pollen related allergy.
2.Role of sufentanil in neural stem cells transplantation for spinal cord injury
Houyong YAN ; Lengmei SONG ; Yue LIU ; Guang YAO ; Ruyi ZHANG
Chinese Journal of Tissue Engineering Research 2017;21(25):4050-4056
BACKGROUND: Studies have shown that the combined use of sufentanil and neural stem cell (NSC) transplantation can increase the number of newborn nerve fibers.OBJECTIVE: To investigate the effect of sufentanil on the hind limb function of rats with spinal cord injury after neural stem cell transplantation.METHODS: (1) Eighty adult female Sprague-Dawley rats were used to build spine cord injury model according to the modified Allen's method and divided into model group, sufentanil group, NSCs transplantation group and sufentanil combined with NSCs transplantation group (combined group). Extra 20 adult female Sprague-Dawley rats were not conducted any treatment as normal control group. (2) After 6 days of modeling, the model rats were subjected to subarachnoid injection of 10 μL of NSC culture medium and intraperitoneal injection of 100 μL of saline in the model group; subarachnoid injection of 10 μL of NSC culture medium and intraperitoneal injection of 100 μL of sufentanil (150 μg/kg) in the sufentanil group; subarachnoid injection of 10 μL of 1×1010/L NSCs suspension and intraperitoneal injection of 100 μL of saline in the NSCs transplantation group; and subarachnoid injection of 10 μL of 1×1010/L NSCs suspension and intraperitoneal injection of 100 μL of sufentanil (150 μg/kg) in the combined group. (3) After 72 hours of modeling, the AQP4 and MMP9 gene expression levels were detected by RT-PCR, and the cell apoptosis changes around the spine cord injury area were determined with TUNEL staining method. (4) The motor functions of rats were tested by Basso, Beattie and Bresnahan score and inclined plane test after 1, 3 days and 1, 2, 3 and 4 weeks of modeling. (5) After 4 weeks of modeling, the histopatholgical changes in the area of spine cord injury were observed by hematoxylin-eosin staining method. The survival changes of NSCs labeled by CM-Dil were determined by fluorescence microscope. The regenerations and distributions of spinal nerve fibers were observed by fluorescein gold retrograde tagging.RESULTS AND CONCLUSION: (1) After 72 hours of modeling, the AQP4 and MMP9 gene expression levels as well as the cell apoptotic rate in the combined group were significantly lower than those in the model, sufentanil and NSCs transplantation groups (P < 0.05). (2) After 2 weeks of modeling, the combined treatment significantly improved the hind limb motor functions of rats compared with the sufentanil and NSCs transplantation groups (P < 0.05), and the recovery of motor function was better in the sufentanil and NSCs transplantation groups than in the model group (P < 0.05). (3) After 4 weeks of modeling, the results of hematoxylin-eosin staining manifested that the spinal cord tissues lost and the magnified syringomyelias occurred in the model group. The syringomyelias in the sufentanil and NSCs transplantation groups were significantly smaller than that in the model group, and the syringomyelias almost disappeared in the combined group. (4) The number of positive cells was the most in the combined group, more in the NSCs transplantation group, but there were no positive cells labeled by CM-Dil in the sufentanil and negative control groups. (5) The number of positive neural fibers in the combined group was the highest followed by the sufentanil and NSCs transplantation groups, and the number of positive neural fibers in negative control group was the lowest. To conclude, sufentanil can improve the recovery of hind limb motor function by reducing the AQP4 and MMP9 expression levels in the injury area, promoting the survival of transplanted NSCs, and decreasing the local NSCs apoptosis after spinal cord injury.
3.Pathology of diabetic myocardiac microangiopathy
Yiqun YAO ; Yanmei CHEN ; Guang LIU ; Zhiyong PEI
Journal of Geriatric Cardiology 2009;6(2):79-81
Objective To investigate the cause of high cardiovascular lethality in patients with diabetics mellitus. Methods Sections from autopsied coronary arteries and myocardium of dead patients with non-insulin-dependent diabetics mellitus and 12 dead control subjects were used for histomorphometric studies. Results The coronary atherosclerotic lesion in diabetics patients was not different in severity from those in controls. Nor was there difference in number of myofibers or diameters of myocardic fibers and capillaries.But the capillary density and the ratio of capillary number to myocardic fiber number in diabetics group were significeantly reduced compared with control group(P<0.0 l),and the capillary basement membrane in the former was significantly thicker than in the latter(P<0.01).Conclision The decrease in number of capillaries and the thickening of basement membrane enhance myocardiac vulnerability to further ischemia and hypoxia,which may undelie high lethality of myocardiopathy in diabetic patients.
6.Study on the susceptible factors of atherosclerotic stenosis in proximal of the coronary artery myocardial bridges
Jian CHEN ; Ping LI ; Ming LIU ; Guang YAO ; Zhengdong WANG ; Peng WU
Chinese Journal of Postgraduates of Medicine 2009;32(28):6-9
Objective To study the susceptible factors of atherosclerotic stenosis before the coronary artery myocardial bridges.Methods The data from 88 myocardial bridge cases which received coronary angiography were statistically analyzed.Sixty-seven cages which suffered from atherosclerofic stenesis in proximal of the myocardial bridges were recruited into group A,and the other 21 cases which suffered no atherosclerotic stenosis or from atherosclerotic stenosis in distal of the myocardial bridges were mcmited into group B.Difference of the age,gender,length of myocardial bridge,systolic blood pressure (SBP),diastolic blood pressure (DBP),pulse pressure (PP),the oppression degree of myocardial bridge (Nobel classification),fasting plasma glucose (FPG),and blood fat,and so on,in two groups,were observed and statistically analyzed.Results The difference of the Nobel classification,SBP and PP in two groups showed a statistical significance (P<0.05).While the difference of the age,gender,length of myocardial bridge,DBP,FPG,total cholesterol,low-density hpoprotein in two groups showed no statistical significance (P>0.05).A further regression analysis suggested that Nobel classification and PP had a correlation with the comphcation of stonosis before the myocardial bridge (r=3.0569,0.9740,P<0.05).Conclusions High blood pressure cases are liable to suffer from myocardial bridge.Myocardial bridges themselves trend to promote or accelerate the atherosclerotic stenosis of the coronary arteries before.them.The oppression degree of myocardial bridge and PP has a correlation with the complication of stenosis before the myocardial bridge,while has no correlations with age,gender,bloodfat,SBP,DBP,FPG,length of myocardial bridge,and so on.
7.A meta-analysis of the association between thyroglobulin gene polymorphism and autoimmune thyroid disease
Jia LIU ; Yuan XU ; Xia GAO ; Zhi YAO ; Pengrui ZHANG ; Guang WANG
Chinese Journal of Endocrinology and Metabolism 2014;30(3):221-224
The association between thyroglobulin gene polymorphism and autoimmune thyroid disease was investigated.A total of eight case-control studies were included.E33SNP was associated with increased susceptibility of autoimmune thyroid disease in recessive model (OR =1.54,95 % CI 1.21 ~ 1.97,P =0.024),while E12SNP was associated with increased susceptibility of autoimmune thyroid disease in allele model (OR =1.12,95% CI 1.05 ~1.21,P =0.033).Thyroglobulin gene polymorphism seems to be associated with autoimmune thyroid disease.
8.Application of serious game learning method in Chinese medical history
Youliang HUANG ; Renquan LIU ; Shuangqing ZHAI ; Guang CHEN ; Qiongjie YAO ; Xing ZHAI
Chinese Journal of Medical Education Research 2014;(9):909-911
“Mechanical memory” approach was commonly used in the knowledge learning of traditional Chinese medicine, which makes students often feel that the knowledge is too abstract, and this method is thus ineffective. “Qihuang the way” using Unity 3D game development platform inte-grates the elements of medical knowledge of the Chinese medical history, and combines TCM theoretical knowledge learning with serious games , which can help invoke students' enthusiasm and improve learning efficiency in an entertaining way.
9.Outward versus inward placement in Shang Ring circumcision for phimosis and redundant prepuce in adult men: analysis of 527 cases.
Sheng-Tao XIE ; Guang-Yu CHEN ; Qiao-Hong WEI ; Xiao-Tong LIU ; Lin JIAO ; Yao TANG
National Journal of Andrology 2014;20(4):325-328
OBJECTIVETo observe the clinical effects of two different circumcision procedures with the Shang Ring and compare their advantages and disadvantages.
METHODSA total of 527 adult males with phimosis or redundant prepuce underwent Shang Ring circumcision by conventional outward replacement (n = 254) and inward placement (n = 273), respectively. We observed the in-ring nocturnal pain, complications, ring-removal pain, degree of edema, recovery time, and patients' satisfaction after surgery, and compared them between the two groups.
RESULTSCompared with the conventional outward placement (5.9%) of the Shang Ring, the inward placement method showed the advantages of mild in-ring nocturnal pain, a low complication rate, significantly reduced ring-removal pain, and mild edema, but exhibited longer healing time.
CONCLUSIONIn Shang Ring circumcision for phimosis and redundant prepuce in adult males, each of the outward and inward placement methods has advantages and disadvantages of its own, but the latter is more advantageous and feasible.
Adult ; Circumcision, Male ; adverse effects ; instrumentation ; Edema ; etiology ; Foreskin ; abnormalities ; surgery ; Humans ; Male ; Pain, Postoperative ; Patient Satisfaction ; Penile Diseases ; etiology ; Penis ; abnormalities ; surgery ; Phimosis ; surgery ; Prostheses and Implants ; Wound Healing
10.Effect of palmatine inhibiting migration,invasion and epithelial mesenchymal transformation in human oral cancer KB cells
Xue-Yun CHENG ; Guang-Yao HU ; Hong-Li LIU ; Chen-Guang LIU ; Yuan-Li DING ; Hui-Ning YANG ; Yi-An ZHAO ; Zhi-Guang SUN
The Chinese Journal of Clinical Pharmacology 2024;40(12):1749-1753
Objective To investigate the effects of palmatine on migration,invasion and epithelial mesenchymal transformation(EMT)in human oral cancer KB cells.Methods KB cells were divided into control group and palmatine-L,-M,-H groups,cultured with 0,4,8 and 16 μmol·L-1 palmatine.After incubation for 48 h,scratch assay was used to detect migration;Traswell assay was used to detect invasion;matrix metalloproteinase 2(MMP-2),MMP-9 and fibronectin(FN)contents were detected by enzyme-linked immunosorbent assay;the expression of Vimentin,N-cadherin and E-cadherin mRNA were detected by real-time quantitative polymerase chain reaction;the expression of Vimentin,N-cadherin,E-cadherin,Wnt3 and β-catenin protein were detected by Western blot.Results Cell mobility in control group and palmatine-L,-M,-H groups were(69.27±8.62)%,(52.94±4.49)%,(45.22±5.05)%and(37.63±4.88)%;the number of transmembrane cells were 197.33±20.26,125.33±12.01,97.00±9.17 and 62.67±7.51;the content of MMP-2 were(2.93±0.21),(1.49±0.13),(1.16±0.15)and(0.95±0.09)ng·mL-1;the content of MMP-9 were(3.51±0.36),(2.37±0.23),(2.06±0.35)and(1.72±0.16)ng·mL-1;the content of FN were(41.28±4.02),(24.03±3.17),(20.67±2.63)and(13.82±2.19)ng·mL-1;the above indexes in palmatine-L,-M,-H groups were compared with the control group,and the differences were statistically significant(P<0.05,P<0.01).The mRNA and protein expressions of Vimentin,N-cadherin and E-cadherin,and the expressions of Wnt3 and β-catenin protein in palmatine-L,-M,-H groups were statistically significant compared with those in control group(P<0.05,P<0.01).Conclusion Palmatine can inhibit the migration,invasion and EMT of human oral cancer KB cells,and its mechanism is related to the regulation of Wnt/β-catenin signaling pathway.