1.RESPONSE OF ASTROCYTES IN CENTRAL VISUAL BRAIN REGIONS OF MONOCULARLY AND BINOCULARLY DEPRIVED RATS
Zhijun YANG ; Li DUAN ; Liangwei CHEN ; Zhiren RAO
Acta Anatomica Sinica 2002;0(05):-
Objective Monocular deprivation(MD) and binocular deprivation(BD) were used to examine the experience\|dependent structural plasticity of astrocytes in the central visual brain regions of young rats. Methods Pups eyelids were monocularly or binocularly sutured on postnatal day 7(P7) and maintained until 80?d,while an unoperated light experienced group was used for comparisions(L).Immunohistochemical ABC method was employed to investigate the immunoreactive changes of astrocytic bodies and processes for glial fibrillary acidic protein(GFAP) in the central visual brain regions. Results The results showed that the amount of astrocytic bodies and processes in the brain was decreased both in MD and BD groups compared to L group.The amount of GFAP\|positive immunoreactivity in the optic chiasma,optic tract,and contralateral central visual brain regions (including suprachiasmatic nucleus,lateral geniculate nucleus,occipital visual cortex,optic nerve layer of the superior colliculus and pretectal area) was significantly decreased in MD group.GFAP\|positive structures presented complementary distribution in bilateral visual cortex.For bilateral brain regions mentioned above,the amount of GFAP\|positive structures basically disappeared in BD group.There was,however,an increase in the olfactory cortex of GFAP immunoreactive in MD and BD groups. Conclusion\ The data suggested that the structure of astrocytes might be influenced by visual experience during development.
2.Stress distribution in natural maxillary central incisor and implant: a three-dimensional finite element analysis
Linzi WAN ; Lei SUN ; Xifeng WU ; Zhenyan LIN ; Liangwei XU ; Zhenzhe LIU ; Duoling XU ; Yang LI ; Yanmin ZHOU ; Yongbo GAO
Chinese Journal of Tissue Engineering Research 2015;19(16):2545-2550
BACKGROUND:Biomechanical compatibility is the necessary condition to ensure the stable osseointegration with implants that then can function over a long period; therefore, it is especialy important to get knowledge about distribution of stress and strain between the maxilary central incisor and its surrounding bone tissue. OBJECTIVE: Based on five different anatomical types of natural teeth, to study the regularity of stress distribution between the maxilary central incisor root and implant.METHODS: According to the five different anatomical types of natural maxilary central incisors, UGNX and ANSYS were used to set up three-dimensional finite element models (B1, B2, M1, M2, P1) for the implant and surrounding structures, which were under 100 N static load at angles of 0o, 30o, 45o, 60o, 90o with the long axis of teeth. Then, the stress distribution between the five kinds of maxilary central incisor roots and implants was analyzed. RESULTS AND CONCLUSION:Among the five different anatomical types, the equivalent stress for both the natural central incisor and implant were increased with the increasing of angles, and the implant had a higher raising trend. The equivalent stress for the natural tooth concentrated upon B1 for the maximum value and M1 for the minimum value; while the equivalent stress for the implant focused on the maximum value at M1 and the minimum value at M2. There was a gap of 2%-31% between the equivalent stresses for the natural tooth roots and a gap of 4%-21% for the implants. The stress distribution range for the implant was just smaler than that for the natural tooth roots. It implies that the bit force of implant and natural tooth is in positive proportion to the bite angles, and the bite force that implant can burden is smaler than that the central incisor can.
3.Distribution character of functional unit of reaction neuron and astrocyte in medullary visceral zone 1 hour after epileptic attack: a laser confocal microscopic research
Zhijun YANG ; Ruxiang XU ; Zhiren RAO ; Ling WEI ; Ying WANA ; Li DUAN ; Liangwei CHEN ; Xiaodan JIANG ; Yingqian CAI ; Mouxuan DU
Chinese Journal of Tissue Engineering Research 2005;9(13):235-237
BACKGROUND: It is considered traditionally that epilepsy is a kind of complicated nervous conduct disorder caused by abnormally excited neuron in different area in brain. While the research on the function of astrocyte in epileptic attack is very rare.OBJECTIVE: To study the reaction of neuron and astrocyte in medullary visceral zone after epilepsy induced by pentetrazole in rats.DESIGN: A randomized controlled experimental research.SETTING and MATERIALS: The experiment was done in the Neurosurgery Laboratory of Zhujiang Hospital Affiliated to Southern Medical University and Neuroscience Institute of Fourth Military Medical University of Chinese PLA. Fourteen healthy adult SD rats, weighing 180 - 220 g, clean grade, were provided by the Experimental Animal Center of Fourth Military Medical University of Chinese PLA.INTERVENTIONS: Distribution of neuron and astrocyte in MVZ 1 hour after epileptic attack was shown by laserconfocal microscopic technique combined with triplication immunofluorescence histochemistry of anti-Fos protein, anti-tyrosine hydroxylase(TH) and anti-glial fibrillary acidic protein(GFAP).MAIN OUTCOME MEASURES: Observation of distribution of positive cells of Fos, GFAP and TH in MVZ and relationship between GFAP positive astrocyte and neuron.RESULTS: Fos positive neurons and GFAP positive astrocytes in MVZ increased significantly. Triplication immunofluorescence histochemistry showed reaction neuron(Fos positive) closely related with reaction astrocyte(GFAP positive) . Three kinds of N-ASC compounds with different labels were found, which were TH +/Fos +/GFAP + three labeled compound, TH + /GFAP +/Fos- and Fos+/GFAP +/TH- two labeled compound.CONCLUSION: Neuron and astrocyte in MVZ reacted strongly when epilepsy attacks. N-ASC as a functional unit may regulate onset of epilepsy.
4.Experimental study on the catecholaminergic effects of paraventricular nucleus projected from medullary visceral zone in "immune-to-brain communication" access
Zhijun YANG ; Ruxiang XU ; Zhiren RAO ; Ling WEI ; Liangwei CHEN ; Li DUAN ; Xiaodan JIANG ; Yingqian CAI ; Mouxuan DU
Chinese Journal of Tissue Engineering Research 2005;9(5):212-214
BACKGROUND: Lipopolysaccharide(LPS), as a polyclonal immune exciter, can simulate immune excitation status, which is useful in the observation of whether the catecholaminergic neurons in paraventricular nucleus of hypothalamus(PVN) projected from medullary visceral zone(MVZ) react towards LPS stimulation that is to provide a theoretical gist for the researches on the protection of brain function.OBJECTIVE: To observe whether PVN catecholaminergic neurons projected from MVZ react towards LPS stimulation for the exploration of the impacts of MVZ-PVN catecholaminergic access in "immune-to-brain communication".DESIGN: A randomized controlled study using experimental animals as subjects.SETTING: An Institute of Neurosurgery and Neurology of one Military University of Chinese PLA.MATERIALS: The study was conducted in the Department of Neurosurgery of Zhujiang Hospital affiliated to Southern Medical University and the Institute of Neurology of the Fourth Military Medical University of Chinese PLA from January to December in 2002. Ten healthy adult SD rats in cleanness grade were obtained from the experimental animal center of the Fourth Military Medical University of Chinese PLA.METHODS: WGA-HRP was injected into PVN of one side of the rat, and the immune exciter LPS was injected into the abdominal cavity after 48 hours of survival to induce immune response. Samples were stained by triple labels of WGA-HRP method and double immunohistochemical staining of anti-Fos and anti-TH antibodies.MAIN OUTCOME MEASURES: To observe the distributions and expressions of WGA-HRP labeled cells, Fos protein, and catecholaminergic neuron(labeled by TH) in MVZ.RESULTS: Seven immune-reactive(IR) positive neurons were found in MVZ, i. e., HRP, Fos or TH single labeled cells, Fos/HRP, Fos/TH or HRP/TH double labeled cells, and Fos/HRP/TH triple labeled cells. Fos/HRP double labeled neurons and Fos/HRP/TH triple labeled neurons accounted for 12. 5% and 39.6% of HRP labeled cells respectively.CONCLUSION: MVZ reacts to LPS immune stimulation, which could upload the immune message to PVN through Catecholaminergic neurons.MVZ might be a relay station in "immune-to-brain communication", which exerts immune modulatory impact through "MVZ→PVN" access.
5.Enhanced recovery after surgery in total laparoscopic radical gastrectomy
Yu YU ; Moucheng ZHANG ; Kaijun GAO ; Liangwei YANG ; Jiaming ZHOU ; Zhilong YAN
Chinese Journal of General Surgery 2020;35(1):21-25
Objective To investigate the effect of enhanced recovery after surgery (ERAS) on immune function and postoperative recovery in gastric cancer patients undergoing total laparoscopic radical gastrectomy.Methods Patients were randomly divided into ERAS group and control group.Blood CD4 +,CD8+,CD4 + CD25 +,C-reactive protein,postoperative recovery and complications were compared between the two groups.Results On day1,CD4 +,CD8 +,CD4 + CD25 + in the two groups were significantly lower than those before surgery (t =9.070,7.297,5.830,12.870,3.529,10.547,all P<0.05).The ERAS group had higher CD8 +,CD4 + CD25 + levels than the control group (t =2.163,2.203,P < 0.05).On day3,CD4 + CD25 + in ERAS group was not different from that before surgery (t =1.062,P > 0.05) while the other indexes in the two groups raised but still were lower than preoperative level (t =3.322,5.015,3.418,9.912,all P <0.05);CD4 +,CDs +,CD4 + CD25 + in ERAS group were higher than control group (t =2.804,2.040,2.210,all P<0.05).On day5,CD4+,CD4 + CD25+ in the two groups and CDs+ in ERAS group returned to the preoperative level,while CDs + of the control group was still lower than the preoperative level (t =6.862,P <0.05).On day1,3 and 5,the C-reactive protein levels of the two groups were higher than those before surgery(t=-13.338,-13.715,-11.319,-12.286,-13.182,-15.076,all P < 0.05),and ERAS group were lower than the control group (t =-3.246,-2.100,-2.211,all P<0.05).There was no mortality in neither groups.The time of passage gas by anus,defecation,getting out of bed,oral feeding,and postoperative hospital stay in the ERAS group were less than thoseinthecontrolgroup[(2.8±1.0)dvs.(3.9±0.9)d,t=-5.974;(3.8± 0.9)d vs.(4.3±1.0)d,t=-2.700;(19.1 ±4.0)hvs.(35.9±6.6)h,t=-16.045;(9.9 ±1.6)d vs.(11.5±2.0) d,t =-4.479,all P < 0.05].Conclusions ERAS in the perioperative period of patients with total laparoscopic radical gastrectomy mitigates the stress on the cellular immune system,reduces inflammatory response,and help fast recover the postoperative gastrointestinal function.
6.A dynamically evolving war between autophagy and pathogenic microorganisms.
Qianqian ZHENG ; Liangwei DUAN ; Yang ZHANG ; Jiaoyang LI ; Shiyu ZHANG ; Hui WANG
Journal of Zhejiang University. Science. B 2022;23(1):19-41
Autophagy is an intracellular degradation process that maintains cellular homeostasis. It is essential for protecting organisms from environmental stress. Autophagy can help the host to eliminate invading pathogens, including bacteria, viruses, fungi, and parasites. However, pathogens have evolved multiple strategies to interfere with autophagic signaling pathways or inhibit the fusion of autophagosomes with lysosomes to form autolysosomes. Moreover, host cell matrix degradation by different types of autophagy can be used for the proliferation and reproduction of pathogens. Thus, determining the roles and mechanisms of autophagy during pathogen infections will promote understanding of the mechanisms of pathogen‒host interactions and provide new strategies for the treatment of infectious diseases.
Autophagy
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Bacteria
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Host-Pathogen Interactions
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Lysosomes
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Signal Transduction
7. Mitochondrial damage induced by HTLV-1 infection in host cells
Xue YANG ; Yecheng XIE ; Yilin GUO ; Xuelu LI ; Huandi LIU ; Liangwei DUAN ; Yuna NIU
Chinese Journal of Microbiology and Immunology 2019;39(12):898-903
Objective:
To investigate the effects of human adult T lymphoblastic leukemia virus typeⅠ (HTLV-1) infection on the production of reactive oxygen species (ROS) and mitochondrial damage in host cells.
Methods:
A cell model of HTLV-1 infection was established by co-culturing HTLV-1-positive cell line MT2 with HeLa cells. ROS, mitochondrial membrane potential (MMP) and total mitochondria were detected using specific fluorescence probe labeling method. Cell apoptosis was detected by Annexin V-FITC/PI method. Western blot was performed to detect viral proteins Tax and p19, as well as mitochondrial proteins TIM23 and TOM20. After the treatment of MT2 cells with different concentrations of reverse transcription inhibitors (ZDV), relative viral loads were detected by quantitative real-time PCR and Western blot, and the mass of mitochondria was analyzed by flow cytometry.
Results:
After co-culturing HeLa cells with MT2 cells for 24 h, the ROS level in host cells increased without obvious cell apoptosis, while the mitochondrial membrane potential, mitochondrial protein expression and total mitochondria decreased significantly. When the replication of HTLV-1 in MT2 cells was inhibited by ZDV, the ROS level and total mitochondria increased.
Conclusions
HTLV-1 infection can cause oxidative stress in host cells, resulting in mitochondrial damage. Autophagy might be activated to degrade mitochondrial damage and maintain cell homeostasis during the infection.
8.Cloning, expression and purification of fructose-2, 6-bisphosphatase gene CpF2KP in papaya.
Liping ZUO ; Qiuxia ZENG ; Xiaobing ZHAO ; Liyuan YANG ; Liangwei XU ; Juan LAI ; Jingjing YUE
Chinese Journal of Biotechnology 2023;39(2):614-624
Papaya, which is mainly cultivated in the southeastern region of China, is one of the four famous fruits in Lingnan. It is favored by people because of its edible and medicinal value. Fructose-6-phosphate, 2-kinase/fructose-2, 6-bisphosphatase (F2KP) is a unique bifunctional enzyme with a kinase domain and an esterase domain that catalyzes the synthesis and degradation of fructose-2, 6-bisphosphate (Fru-2, 6-P2), an important regulator of glucose metabolism in organisms. In order to study the function of the gene CpF2KP encoding the enzyme in papaya, it is particularly important to obtain the target protein. In this study, the coding sequence (CDS) of CpF2KP, with a full-length of 2 274 bp, was got from the papaya genome. The amplified sequence of full-length CDS was cloned into the vector PGEX-4T-1 which was double digested with EcoR I and BamH I. The amplified sequence was constructed into a prokaryotic expression vector by genetic recombination. After exploring the induction conditions, the results of SDS-PAGE showed that the size of the recombinant GST-CpF2KP protein was about 110 kDa. The optimum IPTG concentration and temperature for CpF2KP induction were 0.5 mmol/L and 28 ℃, respectively. The purified sin[A1] gle target protein was obtained after purifying the induced CpF2KP protein. In addition, the expression level of this gene was detected in different tissues, and showed that the gene was expressed at the highest level in seeds and the lowest in pulp. This study provides an important basis for further revealing the function of CpF2KP protein and studying the involved biological processes of this gene in papaya.
Humans
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Carica/genetics*
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Recombinant Proteins
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Carbohydrate Metabolism
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Cloning, Molecular
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China