2.Cloning and expression of Dengue virus nonstructural protein 4A gene and affinity purification of its interacting proteins
Jun XIA ; Jiong XIE ; Peifen ZHANG ; Yuye LI ; Chao LIU ; Xi HUANG ; Ping ZHANG
Chinese Journal of Microbiology and Immunology 2013;(3):184-187
Objective To clone and express Dengue virus nonstructural protein 4A (NS4A) gene and express in eukaryotic cells.Then,to isolate and purify and isolate cellular proteins interacted with NS4A.Methods With specific primers,NS4A gene fragment tagged with FLAG and HA (FLAG-NS4A-HA) was amplified by PCR and cloned into an expression vector,pSG5 vector.Recombinant plasmid was transfected into A549 cells by LipofectAMINETM2000.Transient expression of FLAG-NS4A-HA was detected by Western blot.The NS4A interacting proteins were isolated and purified by tandem affinity purification (TAP) system using HA and FLAG antibodies,and then assayed by silver stained SDS-PAGE.Results Dengue virus NS4A gene tagged with FLAG and HA was successfully constructed into pSG5 vector and expressed in A.549 cells.Silver stained SDS-PAGE showed that the expressed NS4A and two potential interacting proteins that interact with NS4A were isolated after TAP purification and SDS-PAGE.Conclusion Cellular proteins that potentially interacted with Dengue virus NS4A were successfully purified and isolated,which provided a basis for further research.
3.Posterior atlantoaxial fusion fixation for old atlantoaxial injury
Honglin PI ; Peng YU ; Jiakuang LIU ; Jiwen HE ; Qunhai WU ; Chao ZHANG ; Jun ZHANG ; Yan XIE
Chinese Journal of Trauma 2012;(10):926-930
ObjectiveTo investigate the clinical effects of posterior atlantoaxial fusion fixation in treatment of old atlantoaxial injury secondary to atlantoaxial dislocation.MethodsA retrospective analysis was carried out on 16 patients ( 14 males and 2 females) with old atlantoaxial injuries secondary to atlantoaxial dislocations managed with posterior atlantoaxial fusion fixation from March 2008 to March 2012.The time from injury to operation lasted for 3-36 months ( average 10.5 months).Posterior atlantoaxial transpedicular fixation was performed in 13 patients including 10 patients with old odontoid fractures and three with old traumatic transverse ligament disruptions of the atlas combined with atlantoaxial dislocations.Also,posterior atlantal arch transpediclar fixation combined with axial pedicle screw fixation was performed in three patients who had old odontoid fractures combined with atlantoaxial dislocations.All patients had simultaneous autogenous bone grafting between atlas and axis during reduction and fixation.The preoperative and postoperative Japanese Orthopaedic Association (JOA) scores were compared.Follow-up X-ray films and CT was performed to evaluate the atlantoaxial reduction and fusion.ResultsAll the patients were followed up for 9-18 months ( mean 13 months).None of the patients had spinal cord or vertebral artery injuries.Follow-up CT showed that two patients had partial penetration of one side axial pedicle screws into transverse foramen without nerve and blood vessel injuries.Clinical symptoms obtained different degree of improvement.The postoperative JOA scores ranged from 13 to 16 points ( mean 14.8 points) and the improvement rate of JOA was 71%-92% ( mean 82% ).The X-ray films and CT showed sound bone fusion,with good location of screws but with no signs of atlantoaxial instability or loss of reduction,or loosening or breakage of the screws.ConclusionPosterior atlantoaxial fusion fixation can effectively reconstruct atlantoaxial stability,improves neurologic function of spinal cord and has reliable curative effects.
4.Influence of protein phosphatase 5 on the lipid metabolism in mice
Jun WANG ; Bin QIU ; Ming LIU ; Chao WANG ; Weidong YONG ; Zhongwen XIE
Chinese Journal of Comparative Medicine 2016;26(8):79-84
Objective To investigate the effect of protein phosphatase 5 (PP5) on lipid metabolism in the PP5 knockout (KO) mice.Methods Male PP5 KO and wild type (WT) mice at the age of 6 weeks were used in this study. In order to study the effect of high fat diet ( HFD) feeding, the body weight was measured.The liver histology was examined by HE and oil red O staining.To further verify PP5 functions in the adipogenesis, in vitro experiment was carried out using mouse embryonic fibroblasts (MEF).Western blotting and real-time PCR were performed to quantified the expression of lipid metabolism-related genes in the liver tissues.Results Compared with the WT mice, the body weight gain was slower in the KO mice.The size of the lipid droplets was smaller and the quantity was less in the KO mouse liver tissue.In vitro study revealed that the KO mouse MEF cells showed less differentiated adipocytes with smaller lipid droplets than the WT MEF cells.This observation was further confirmed by detecting the expression of adipogenesis-related genes in the HFD liver.The markers of adipocyte differentiation, such as CD36, AP2, PPARγ2, and Glut4, were significantly decreased, while energy expenditure-related markers, such as phosphorylation of GR and expression of UCP1, were significantly increased.Conclusions Protein phosphatase 5 may play a regulatory role in the mouse lipid metabolism through regulating the de-phosphorylation of p-GR and enhancing the expression of UCP1.
5.Clinical evaluation study on long-term effect of acupuncture with pattern/syndrome differentiation on functional dyspepsia.
Duo-Duo LI ; Zeng-Hui YUE ; Li-Chao XU ; Tao XIE ; Gang-Zhu HU ; Jun YANG
Chinese Acupuncture & Moxibustion 2014;34(5):431-434
OBJECTIVETo compare the efficacy difference in the treatment of functional dyspepsia between acupuncture at the acupoints selected by pattern/syndrome differentiation and domperidone.
METHODSSeventy cases were randomized into an acupuncture group (35 cases) and a western medication group (35 cases). In the acupuncture group, Zusanli (ST 36) and Neiguan (PC 6) were selected. Taichong (LR 3) and Neiting (ST 44) were added for excess syndrome while Gongsun (SP 4) and Yinlingquan (SP 9) were added for deficiency syndrome. A pair of electrodes was attached to one acupoint and an assistant point (2 mm next to the acupoint centripetally) and stimulated with disperse-dense wave at 2 Hz/100 Hz, once a day. In the western medication group, domperidone was prescribed for oral administration, 10 mg each time, three times a day. In the two groups, the treatment of 5 days made one session and 4 sessions were required totally. Nepean dyspepsia index (NDI) was compared after treatment, 1, 2, 3, 4 and 5 months after treatment between the two groups respectively.
RESULTSThe score of symptom and score of life quality in NDI after treatment and at each follow-up time point were improved obviously in the acupuncture group as compared with those before treatment (all P < 0.01). In the western mediation group, the score of symptom and the score of life quality in NDI after treatment and in follow-up of 1, 2 and 3 months were improved obviously as compared with those before treatment (all P < 0.01), but the differences were not significant in follow-up of 4 and 5 months (both P > 0.05). Compared with the western medication group, the symptom score of NDI was reduced obviously after treatment and in each time point of follow-up in the acupuncture group (P < 0.05, P < 0.01), and the score of life quality was increased obviously (P < 0.05, P < 0.01).
CONCLUSIONAcupuncture at the acupoints selected by pattern/syndrome differentiation and domperidone are effective in the treatment of functional dyspepsia. Domperidone is unsatisfactory in the long-term effect, but acupuncture achieves the positive short-term and long-term effects on functional dyspepsia.
Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Dyspepsia ; diagnosis ; psychology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Time ; Treatment Outcome ; Young Adult
6.Endothelial dysfunction in young pre-hypertension and its influencing factors
Lan WANG ; Jianjun MU ; Ruihai YANG ; Jun YANG ; Man WANG ; Chao CHU ; Bingqing XIE ; Zhenzhen DONG
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(4):574-578,625
Objective To investigate the endothelial dysfunction in pre-hypertension and its influencing factors.Methods A total of 373 youth were divided as the subjects into hypertension group (HBP group),prehypertension group (PHT group) and normal blood pressure group (NBP group).Endothelial function was assessed based on carotid intima-media thickness (IMT),brachial artery flow-mediated dilation (FMD) and brachial-ankle pulse wave velocity (baPWV).Results IMT and baPWV in PHT group were higher than those in NBP group (P<0.05),but did not reach the significant difference when compared with HBP group (P>0.05).Compared with HBP,the levels of FMD in PHT group significantly increased (P< 0.05);however,no difference was observed in comparison with NBP group (P>0.05).In the early stage of hypertension,diastolic BP (β=-0.120,P<0.05) and body mass index (β=-0.115,P<0.05) were negatively correlated with FMD;diastolic BP (β=0.146,P<0.05),2-hour glucose (β=0.147,P<0.05),high-density lipoprotein cholestrol (β=0.150,P<0.05),and waist-hip ratio (β=0.126,P<0.05) showed a positive correlation with IMT.baPWV was correlated with systolicBP (β=0.358,P<0.01),waist circumference (β=0.254,P<0.05),fasting glucose (β=0.155,P<0.05),postprandial 2 h blood glucose (β =0.152,P <0.05),uric acid (β =0.206,P < 0.05),and C-reactive protein (β=0.099,P<0.05).Corclusion Our study shows that endothelial dysfunction may exist in the prehypertensive young,and several cardiovascular risks contribute to its development in the early stage of hypertension.
7."Treating Diabetes Mellitus from the Viewpoint of Spleen: Based on the Theory of ""Homogeny of Spleen and Pancreas"""
Jun CHAO ; Ya LIU ; Wen ZHONG ; Chuanming FANG ; Chunguang XIE ; Hong GAO
Journal of Traditional Chinese Medicine 2017;58(17):1458-1461
The academic origin of homogeny of spleen and pancreas is explained from the aspect of Chinese medicine.The authors think spleen faihng to spread essence is the basic pathogenesis to diabetes mellitus.Spleen function of spreading essence is impaired.Thus essence of water and grain could not be spread in the whole body but amass sugar-turbidity,which manifests high blood sugar.Differentiating diabetes mellitus from the viewpoint of spleen,invigorating spleen and benefiting Qi could help spleen to ascend clear.Invigorating spleen-yin and clearing endogenous heat are used.The liver and kidney should be considered.The methods of dissipating phlegm and activating blood circulation could be combined.The treating idea of treating spleen is treating pancreas should be used in preventing and treating diabetes mellitus.
8.Effects of different cryopreservation methods on the ultrastructure and viability of amniotic membrane
Dai LIU ; Jie JIN ; Fang XIE ; Chao ZHANG ; Jianjian LU ; Jiajie XU ; Jun XU ; Li TENG
Chinese Journal of Tissue Engineering Research 2015;(15):2376-2381
BACKGROUND: There are currently many cryopreservation methods for the aminotic membrane, which have varying effects on the ultrastructure and biological activity of amniotic membrane, but on no one is effective.
OBJECTIVE: To compare the effects of different cryopreservation methods on the ultrastructure and viability of aminotic membrane and to seek the ideal cryopreservation method.
METHODS: Aminotic membrane separated from the fresh placenta was preserved respectively with deep-frozen cryopreservation and vitrification, and everyway was run for 3 and 6 months. Fresh aminotic membrane was used as control. The ultrastructure of aminotic membrane was observed by transmission electron microscopy, and the viability of aminotic membrane was assessed by microcomputer analysis system for biological oxygen consumption, and immunohistochemical staining combined with image analysis system was used for lactate dehydrogenase activity.
RESULTS AND CONCLUSION:After 3 and 6 months of crypreservation, the damage to the ultrastructure of aminotic membrane by vitreous cryopreservation was slighter than that of amniotic membrane cryopreserved at-80℃. Compared with the fresh aminotic membrane, the gray value of lactate dehydrogenase and partial pressure of oxygen were significantly decreased in the cryopreserved aminotic membrane by deep-frozen cryopreservation at 3 and 6 months (P < 0.05) and by vitreous cryopreservation at 6 months (P < 0.05), but there was no statisticaly significant difference in the change rate of oxygen partial pressure and the gray value of lactate dehydrogenase between the fresh aminotic membrane and the cryopreserved aminotic membrane by vitreous cryopreservation at 3 months. The present study led to the conclusion that vitreous cryopreservation protocol alows to not only maintain the integrity of AM, but also to preserve the viability of the cels. So the vitreous cryopreservation is superior to the deep-frozen cryopreservation for cryopreservation of aminotic membrane.
9.Posterior short-segment fixation with undermining decompress for upper lumbar burst fractures.
Mao-sheng ZHOU ; Jia-bing XIE ; Guo-zheng DING ; Qiang WANG ; Zhu-jun XU ; Chao FANG ; Min YANG
China Journal of Orthopaedics and Traumatology 2015;28(12):1132-1136
OBJECTIVETo observe clinical effects of posterior short-segment fixation with undermining decompress by posterior ligament complex for the treatment of upper lumbar burst fractures.
METHODSFrom October 2010 to March 2013,23 patients with upper lumbar burst fractures (Denis B type) were treated by posterior short-segment fixation with undermining decompress by posterior ligament complex. There were 18 males and 5 females aged from 26 to 64 years old with an average of 45.7 years old. Twelve patients were caused by falling down, 5 cases were caused by traffic accident, 4 cases were the bruise injury caused by heavy object and 2 cases were caused by other injury. Fourteen patients were L1 fracture and 9 patients were L2 fracture. Thirteen patients were combined with nerve injuries (degree D according to ASIA classification). Internal fixation were removed from 12 to 20 months with an average of 14.3 months. JOA scores and imaging changes were recorded and compared at different time points.
RESULTSAll patients were followed up from 18 to 24 months with an average of 20.4 months. Thirteen patients with nerve injuries were completely recovered at 3 to 6 months after operation. JOA score at 1 year after operation was 20.63 ± 0.92, and 20.38 ± 1.06 at 3 months after removal of internal fixation,which were improved obviously than 9.90 ± 2.73 at 3 months after operation. (P > 0.05) Anterior height of injured vertebrae, vertebral body angle and local Cobb angle was (95.0 ± 0.53)%, (2.78 ± 1.36) and (2.43 ± 1.52) °respectively, and improved obviously than that of before operation (P < 0.05). There was no statistical significance in JOA scores at 3 months after removal of internal fixation and 1 year after operation (P > 0.05).
CONCLUSIONposterior short-segment fixation with undermining decompress by posterior ligament complex for the treatment of upper lumbar burst fractures has advantages of minimally invasive, could effective recover vertebrae height, maintain stability of spine, decrease low back pain. It is a safe and effective operative method.
Adult ; Decompression, Surgical ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Spinal Fractures ; surgery
10.Effect of erythropoietin on signal transduction pathway in rat model of retinal detachment
Zheng-gao, XIE ; Fang, CHEN ; Chao-rong, ZHUANG ; Jian, WANG ; Hong, WANG ; Ya-kun, WANG ; Jun, ZHU
Chinese Journal of Experimental Ophthalmology 2012;30(2):141-145
BackgroundOur previous study showed that erythropoietin (EPO) protects the photoreceptor from apoptosis in retinal detachment(RD) rat,but its signal transduction pathway remains unknown.Objective The present study was to investigate the effects of EPO on signal transduction pathway in RD.MethodsTwentyfour albino clean Sprague-Dawley(SD) rats were randomly divided into 4 groups.5 μl PBS was injected into vitreous cavity of rats in RD+PBS group,and 400 ng EPO(5 μl) was used at the same way in RD+EPO group.Three days later,the rats were sacrificed and the retina was isolated in each group.The expression levels of Janus kinase 2 (JAK2),p-JAK2,Akt,p-Akt,extracellular regulated protein kinase-1/2 ( ERK-1/2 ),p-ERK-1/2,signal transducer and activator of transcription 5 ( STAT5 ),p-STAT5,nuclear factor-kB (NF-sB) and p-NF-kB were detected by Western blot assay.The administration of experimental animals followed the Standard of ARVO.ResultsThree days after RD,the expression levels of JAK2,Akt and ERK-1,ERK-2 in retinas among normal group,RD,RD+PBS,RD+EPO groups were statistically insignificant different ( F =0.298,P =0.826 ; F =0.681,P =0.588 ; F =0.978,P=0.450;F=1.115,P=0.399 ),but the levels of p-JAK2,p-Akt,p-Erk-1 and p-Erk-2 among these 4 groups were significant difference ( F=24.435,P =0.000; F=48.163,P =0.000;F =19.092,P =0.001; F =14.393,P=0.001 ),and those in RD+EPO group was significantly higher than that in RD and RD+PBS groups( P<0.05 ).The expression levels of STAT5 and NF-kB among the 4 groups were no significantly differences (F =1.136,P=0.391 ;F=0.696,P=0.580),but after the phosphorylation of STAT5 and NF-kB,the differences was significant ( F =14.189,P =0.001 ; F =40.103,P =0.000 ).Those in RD,RD + PBS,RD + EPO groups did not increase either (P>0.05).Although the levels of p-STAT5 and p-NF-kB in RD,RD+PBS,RD+EPO groups were significantly higher than those in normal control group( P<0.05 ),the level of p-STAT5 in RD+EPO group was not significantly higher than that in RD and RD + PBS groups (P > 0.05 ). Conclusions PI3K/Akt and MAPK/ERK-1/2 signal transduction pathways might participate in the protecting process of EPO to photoreceptor in RD rats.