1.Screening and Analysis of Proteins Interacting with HCMV UL145 from a Human Fetal Brain cDNA Library
Fei ZOU ; Shuang WANG ; Si WU ; Zhengrong SUN
Journal of China Medical University 2017;46(4):309-312
Objective To screen a human fetal brain cDNA library for proteins that can interact with HCMV UL145 using a yeast two-hybrid system.Methods A bait plasmid (pGBKT7-UL145) was constructed.Using HCMV UL145 as bait,a human fetal brain cDNA library was screened and proteins interacting with UL145 were identified using bioinformatic methods to sequence and analyze the positive clones.Results Three clones interacting with HCMV UL145 were found,and identified as FOXG1.Conclusion Several proteins interacting with HCMV UL145 in the human fetal brain cDNA library were identified as FOXG1,indicating that this protein may play an important role in the course of HCMV infection.
2.Advance of Mirror Integrated Therapy for Upper Limbs Rehabilitation (review)
Shuang LIANG ; Renling ZOU ; Yabin JIANG ; Xiulin XU ; Xiufang HU
Chinese Journal of Rehabilitation Theory and Practice 2017;23(1):59-62
Mirror therapy has been widely used in practice as an effective rehabilitaiton method. Mirror integrated therapy combined mirror therapy with some other technologies, such as virtual reality, functional electrical stimulation, augmented reality, etc., to enhance the task orientation, and improve the efficiency of treatment.
3.Effects of exercise on behavior and peripheral blood lymphocyte apoptosis in a rat model of chronic fatigue syndrome.
Jun, ZOU ; Jianqi, YUAN ; Shuang, LV ; Jiaheng, TU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(2):258-64
This study examined the effects of exercise on behavior and peripheral blood leukocyte apoptosis in a rat model of chronic fatigue syndrome (CFS). Thirty-six healthy male Sprague-Dawley rats were equally randomized into 3 groups: the control group, CFS model group and the exercise group in terms of body weight. A total of 25 rats entered the final statistical analysis due to 11 deaths during the study. CFS model was established by subjecting the rats in CFS model group and exercise group to electric shock, chronic restraint stress and cold water swim. Besides, rats in the exercise group took running wheel exercise. After a week of conditioning feeding, model construction and running wheel exercise were performed simultaneously, and lasted for 23 consecutive days. The behavior experiments, including running wheel exercise, open-field test, tail suspension test and Morris water maze test, were conducted, either before or after the model establishment. Rats were sacrificed and peripheral blood was obtained for the assessment of lymphocyte apoptosis index by flow cytometry (FCM). It was found that as compared with those in the control group, the weight of the rats was decreased obviously (P<0.01), the mobility time in the open-field and the tail suspension tests was shortened significantly (P<0.01), the time to locate the platform was enhanced (P<0.01) and the cell apoptosis index was increased substantially (P<0.01) in the CSF model group. Meanwhile, in comparison to the model group, the behavior in the open-field and the tail suspension tests was improved significantly (P<0.05), and the apoptosis index decreased remarkably (P<0.01) in the exercise group. It is concluded that sport intervention can prevent lymphocyte apoptosis and improve animal behavior rather than the memory.
4.The influence of acupuncture on the JNK signal transduction pathway in experimental acute intercerebral hemorrhage rats
Fang LIU ; Wei ZOU ; Xiaowei SUN ; Shuang LI
Chinese Journal of Primary Medicine and Pharmacy 2010;17(17):2309-2311
Objective To explore the influence on the JNK signal transduction pathway by BaiHui-QuBin Scalp-acupuncture in rats of experimental acute intercerebral hemorrhage. Methods 192 healthy Wistar male rats were assigned into 3groups randomly:model group, model add pinprick group (to abbreviate:pinprick group), model add suppressor group(to abbreviate:suppressor group) ,containing 60 rats in each. All rats were induced be the intercerebral hemorrage model. Another 12 rats were assigned as the matched blank group. The pinprick group were acupunctured on the "Baihui" penetrating "Qubin". The influence surrounding the hamatoma of the proteinum expression of p-JNK of each group in deferent time phase point with the method of Western blot was observed. Results The results of expression of the proteinum p-JNK with the method of Western blot: all the rats after being made model appeared the phenomenon of the expression of p-JNK of rat's brain tissue surrounding the hematoma and increased to the peak in 2 days,but decreased gradually in 7days. In the same time phase point,compared with the model group,the expression of p-JNK of the pinprick group and the suppressor group were lower obviously (P < 0.01). The pinprick group compared with the suppressor group, the expression of p-JNK showed no significant difference (P > 0.05). Conclusion The therapy of BaiHui-QuBin Scalp-acupuncture could block the JNK signal tranaduction pathway, inhibit the expression of the proteinum p-JNK and so on have the function of protecting the nerve cell and inhibiting apoptosis.
5.Knowledge base, research front and hot spot analysis of Cu-Zn superoxide dismutase
Yu ZOU ; Yanjun LI ; Shuang YANG ; Qingwen ZHANG
Chinese Journal of Tissue Engineering Research 2015;(42):6861-6867
Abstract BACKGROUND:Studies have found that Cu-Zn superoxide dismutase (SOD1) is associated with familial amyotrophic lateral sclerosis, opening up a new way of studying molecular genetics. So far more than 100 kinds of SOD1 gene mutations have been found. OBJECTIVE:To analyze the hot spot, research front and knowledge base of amyotrophic lateral sclerosis visualy. METHODS:Totaly 4 693 relevant articles published from 2005 to 2014 were retrieved from Web of Science in ISI with “Cu-Zn superoxide dismutase” or “SOD1” as search keywords. With the aid of CiteSpace III software, the visualization mapping of the network in co-cited articles and keywords was drawn to reveal knowledge base, hot spots and research front of SOD1. The parameters include the number of published papers and citations within 10 years, distribution of research countries and institutions, main source journals, research area of highly cited papers, keywords with high-frequency and emerging keywords with high-frequency in recent 5 years. RESULTS AND CONCLUSION:The number of published papers and citations in a year showed a trend of sustained growth. United State, China and Japan rank the top three in this area, in which Chinese Academy of Sciences has a great influence among the research institutions. The research fields of SOD1 focus on neurosciences and neurology, biochemistry and molecular biology and so on. The high impact factors of journals with a large number of articles reflect the importance and innovation of this research. Ten high-cited articles consist of the knowledge base on SOD1, directing to the finding of different sites of SOD1 mutation and the measurement of protein concentrations and activity of SOD. The hot spots of SOD1 mainly focus on oxidative stress, familial amyotrophic lateral sclerosis caused by SOD1 mutation and different types of transgenic animal models. The research fronts mainly focus on the finding of pathogenesis in amyotrophic lateral sclerosis, such as the aggregation of TDP-43, the interaction between astrocytes and motor neurons, optineurin and the inhibitor of nuclear factor-κB, hexanucleotide repeat expansion in C9ORF72 and autophagy.
7.Screening and identification of proteins interacting with HCMV UL132 protein
Fei ZOU ; Shuang WANG ; Si WU ; Zhengrong SUN
Journal of Jilin University(Medicine Edition) 2017;43(4):690-693,前插1
Objective:To screen the proteins interacting with the human cytomegalovirus(HCMV)UL132 protein from the human fetus brain cDNA library by using Yeast Two-Hybrid System, and to elucidate the possible mechanism of UL132 protein in congenital cytomegalovirus infection.Methods:The HCMV UL132 fragment was amplified by polymerase chain reaction,the amplified HCMV UL132 fragment and expression vector pGBKT7 were digested and purified,and the HCMV UL132 fragment was linked to the vector pGBKT7.The pGBKT7-UL132 was constructed and transformed to yeast AH109, then the Human Fetal Brain DNA Library DNA was transformed into AH109 yeast.Using HCMV UL132 as abait, a human fetus brain cDNA was screened and the proteins interacting with UL132 protein were searched, the positive clone was sequenced and analyzed by bioinformatics methods.Results:The bait expression vector pGBKT7-UL132 was successfully constructed.The results of double enzyme digestion showed that there were two visible bands of 800 and 7 000 bp, respectively.After transformation of library plasmid, the transformation efficiency was calculated, and the transformation efficiency was 6.6×103 cfu· μg-1.There were 95 blue clones by X-gal coloration reactionsequencing and there were 10 clones interacting with the protein encoded by UL141 protein.The BLAST analysis showed that 7 of them were highly homologous with CAML.Conclusion:CAML might be one interaction protein with HCMV UL132 in Human Fetus Brain cDNA Library,suggesting that the interaction may be associated with the invasion and proliferation of the HCMV.
8.Role of protein kinase C in the activation of store-operated Ca(2+) entry in airway smooth muscle cells.
Yadong, GAO ; Jinjing, ZOU ; Shuang, , GENG ; Junwen ZHENG ; Jiong, YANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(3):303-10
Store-operated Ca(2+) channels (SOCs) are plasma membrane Ca(2+) permeable channels activated by depletion of intracellular Ca(2+) store. Ca(2+) entry through SOCs is known as store-operated Ca(2+) entry (SOCE), which plays an important role in the functional regulation of airway smooth muscle cells (ASMCs). Protein kinase C (PKC) has been shown to have an activating or inhibiting effect on SOCE, depending on cell types and PKC isoforms that are involved. In ASMCs, the effect of PKC on SOCE has not been elucidated so far. In this study, the role of PKC in the activation of SOCE in rat ASMCs was examined by using Ca(2+) fluorescence imaging technique. The results showed that acute application of PKC activators PMA and PDBu did not affect SOCE induced by the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin. The non-selective PKC inhibitor chelerythrine significantly inhibited thapsigargin- and bradykinin-induced SOCE. RT-PCR assay identified PKCα, δ and ɛ isoforms in rat ASMCs. PKCα-selective inhibitor Gö6976 and PKCɛ-inhibiting peptide Epsilon-V1-2 had no effect on SOCE; by contrast, PKCδ-selective inhibitor rottlerin attenuated SOCE dramatically, suggesting that PKCδ was the major PKC isoform involved in the activation of SOCE in ASMCs. Moreover, PKC down-regulation by extended exposure to high doses of PMA or PDBu also reduced SOCE, confirming the essential role of PKC in the activation of SOCE in ASMCs. In addition, PKC down-regulation did not influence the expression of stromal interaction molecule 1 (STIM1) and Orai1, two elementary molecules in the regulation and activation of SOCs. These results identified PKCδ as an essential PKC isoform involved in the activation of SOCE, and confirmed that PKC regulates the function of ASMCs in a SOCE-dependent manner.
9.Effects of down-regulation of protein kinase C on activation of storeoperated Ca~(2+) channels and the proliferation of airway smooth muscle cells
Yadong GAO ; Jinjing ZOU ; Junwen ZHENG ; Shuang GENG ; Jiong YANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM:To investigate the effects of down-regulation of protein kinase C (PKC) on the activity of storeoperated Ca2 + channels (SOC) and the proliferation of airway smooth muscle cells (ASMCs). METHODS:Rat bronchial smooth muscle cells were isolated and cultured. Fluo-3 /AM fluorescence was measured by laser confocal microscope to assessing intracellular Ca2 +. Downregulation of PKC activity was achieved by incubation of ASMCs with PKC activator phorbol-12-myristate-13-acetate (PMA,10 ?mol/L) or phorbol 12,13 -dibutyrate (PDBu,1 ?mol/L) for 24 h. The proliferation of ASMCs was assayed by calculating the reduction rates of Alamar blue. RESULTS:Down-regulation of PKC activity by longterm exposure of PMA or PDBu inhibited the proliferation of ASMCs,the similar results were obtained by using PKC inhibitor chelerythrine. Both downregulation of PKC activity and inhibition of PKC activity by chelerythrine reduced Ca2 + entry through SOC channels. Low concentration of PMA (0. 1 ?mol/L) promoted the proliferation of ASMCs,and this effect was inhibited by SOC blocker SKF-96365. CONCLUSION:Inhibition or down -regu-lation of PKC activity results in the inhibition of SOC channels,suggesting that PKC is involved in the activation of these channels. Ca2 + entry through SOC channels might contribute to PKC-promoted proliferation of ASMCs.
10.The analysis of measurement results of adult myopia biological optical parameters
Ying WANG ; Yunchun ZOU ; Shuang XU ; Yang YANG
Journal of Chinese Physician 2015;(3):328-330,336
Objective To investigate the correlations between the ocular biometric optical parame-ters and refractive dioptres in adults, assess the influence of the biometric optical parameters on the develop-ment of myopia, and provide a reference of prevention and treatment for myopia.Methods A total of 188 adults (376 eyes) aged 18 to 40 years old was enrolled in this study.Dioptres were measured with phorop-ter, A-mode ultrasound was used to measure the vitreous length and lens thickness, the axis length, anterior corneal surface semidiameter anterior chamber depth were measured with intraocular len-master ( IOL-mas-ter) .SPSS 17.0 software was used to analyze the date.Results ⑴Of the 376 eyes, the results showed a statistically significant difference between myopia and emetropia in the axis length, vitreous depth, anterior corneal surface semidiameter, anterior chamber depth and lens thickness( P <0.01).⑵The partial corre-lation analysis indicated a significant positive correlation between the refraction dioptres and axial length, vitreous length ( r =-0.90, -0.88, respectively, P <0.01);and significant strong correlation was also found between dioptres and anterior corneal surface horizontal and vertical semidiameter ( r =0.81, 0.84, respectively, P <0.01);But correlations between dioptres and lens thickness and anterior chamber depth were very weak ( r =-0.11, 0.12, respectively, P <0.01);the correlation between axial length and vit-reous depth was very high ( r =0.95, P <0.01).and the data indicated that the anterior corneal surface horizontal and vertical semidiameter were high positive correlation( r =0.91, P <0.01);the same high positive correlation between the axial length and the anterior corneal surface semidiameter ( horizontal/verti-cal) was also found ( r =0.78,0.81, P <0.01).Conclusions Two chief factors (corneal surface semi-diameter and ocular axial length) influencing the level of dioptres were high positive correlation, seeking the mutual coordination factors which promote corneal curvature change and axial growth and finding out the feedback mechanism of promoting the cornea sphericity might be conducive to further explore how to control the occurrence and development of myopia.