2.Using the SELDI Protein Chip System to Detect Changes in Protein Expression in Vero Cells after Infection
Zhi-jun, LIU ; Bin, WANG ; Zhi-yong, YAN ; Xu-xia, SONG ; Dong-meng, QIAN ; Zhi-qiang, BAI
Virologica Sinica 2007;22(1):68-73
Human herpes simplex virus 1 (HSV-1) causes facial,ocular,and encephalitic disease and is associated with latent infection and cancer.Here,we developed a means of studying the pathogenesis of HSV-1 infection at the protein level by using the SELDI Protein Chip to detect changes of protein expression in Vero cells cultured in vitro.After infection with HSV-1 and culture for 12,24 or 48 h,cells were harvested and lysed.IMAC3 arrays were applied to SELDI-TOF-MS to detect proteomic differences before and after infection.The chip detected a series of differentially expressed protein peaks.Interestingly,both peaks at 16 912 Da and 17 581 Da corresponded precisely with the molecular mass of ISG 15,which may participate in antiviral activity during the process of infection.Thus,the results we obtained can serve as a basis to study the pathogenesis of HSV-1 and the interaction between the virus and its host.In addition,they can help in the discovery of new therapeutic targets for treatment of HSV-1 infection.
3.Protective effect of melatonin on oxidative stress inducing hair follicle injury in scald rat.
Jun ZHANG ; Da-Hai HU ; Gang CHEN ; Xiao-Zhi BAI ; Chao-Wu TANG
Chinese Journal of Burns 2009;25(2):129-132
OBJECTIVETo investigate the protective effect of melatonin on residual hair follicle cells of scald rats at early stage.
METHODSEighteen male Sprague-Dawley rats were randomly divided into scald group, treatment group, sham group , with 6 rats in each group. The rats in scald group and treatment group were subjected to 30% TBSA partial thickness scald on the back, and were resuscitated with balanced solution after 1 hour, while those in sham group were immersed in water at 37 degrees C for 25 s to simulate scald, and did not receive fluid replacement. Rats in treatment group were intraperitoneally injected with 10 mg/kg melatonin solution at 1 minute, 8 hours and 12 hours after scald, while those in sham group and scald group were given equal volume of 1% alcohol sodium-isotonic saline instead. Tissue samples were harvested at 6, 12 and 24 post scald hours (PSH) for determination of MDA and GSH levels. Apoptosis of residul hair follicle was detected by TUNEL method and immunohistochemistry of caspase-3.
RESULTSThe level of MDA in scald group at each time point was much higher than that in sham group (P < 0.01) and treatment group (P < 0.05), and it peaked at 12 PSH. The changes in GSH were just opposite to that of MDA. Under fluorescence microscope, the residual hair follicle cells were blue, and the apoptotic cells appeared green. The apoptosis rate in scald group at 6, 12, 24 PSH was obviously higher than that in sham (P < 0.01) and treatment groups (P < 0.05), which was (20.2 +/- 3.4)% vs (4.3 +/- 2.3)% vs (10.9 +/- 3.2)%, (31.2 +/- 3.6)% vs (5.1 +/- 2.5)% vs (19.1 +/- 3.7)%, (22.4 +/- 2.7)% vs (4.1 +/- 2.4)% vs (13.1 +/- 3.4)%, respectively. The score of caspase-3 positive cell in scald group was higher than those in sham group (P < 0.01) and treatment group (P < 0.05).
CONCLUSIONSThere is obvious correlation between oxidative stress and apoptosis rate of hair follicle cells in rats with partial thickness scald. Early administration of melatonin may have anti-apoptosis ability for residual hair follicle cells by attenuation of oxidative stress.
Animals ; Apoptosis ; Burns ; drug therapy ; metabolism ; Hair Follicle ; cytology ; metabolism ; Male ; Melatonin ; therapeutic use ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley
4.Study on the resource of cytokine gene modified seed cells in bone tissue engineering: the stable expression of fibroblasts after bone morphogenetic protein-3 transfection
Jian LIU ; Guolin MENG ; Yunyu HU ; Zhi YUAN ; Rong Lü ; Jun WANG ; Xinzhi XU ; Jianping BAI
Chinese Journal of Tissue Engineering Research 2005;9(2):226-227
BACKGROUND: Bone morphogenetic protein(BMP) is one of the most important cytokines that induce and promote seed cells to be transformed into osteocytes. Insoluble natural BMP can hardly affect the life of cultured seed cells. The expensive soluble recombinant BMP is also hard to work on the seed cells at the appropriate time and dose. Therefore, gene therapy technique provides us with a brand new idea of using gene-modified seed cells.OBJECTIVE: To transfect exogenous BMP-3 gene into the fibroblasts and screen the positive fibroblast clones that can express BMP-3 stably.DESIGN: Simple sample study.SETTING: Orthopaedic Research Institute, Xijing Hospital, Fourth Military Medical University of Chinese PLA.MATERIALS: The fibroblasts(NIH3T3) were kindly presented by Professor Situ Zhen-qiang of the Stomatological College of Fourth Military Medical University of Chinese PLA.METHODS: This experiment was conducted in the Key Laboratory of Chinese PLA, which belongs to the Orthopaedic Research Institute of Fourth Military Medical University. BMP-3 gene was transfected into the fibroblasts through lipofectamin. The transfected cells were screened by G418. The separated cloned cells were identified through immunohistochemistry. The positively stained cells were the clones of BMP-3 expressing fibroblasts.MAIT OUTCOME MEASURES: The screening concentration of NIH3T3 cells, screening of positive transfected cells, and expression of BMP-3 in screened cells.RESULTS: BMP-3 gene was successfully transfected into the fibroblasts. BMP-3 expressing fibroblast clones were creened and identified through immunohistochemistry. Fibroblast strains with stable BMP-3 expression were obtained.CONCLUSION: The transfection of BMP-3 gene eukaryonic expression vector into the fibroblasts and obtaining of fibroblast strains with BMP-3 expression have laid foundation for the usage of gene-modified seed cells in future research of bone tissue engineering.
5.Identification and the mRNA expression of HS1-associated protein X-1 in the peripheral blood mononuclear cells of patients with systemic lupus erythematosus
Zhi-Fang ZHAI ; Hui-Lin WANG ; Bai-Yu ZHONG ; Ying-Bo WEI ; Jun DENG ; Fei HAO ;
Chinese Journal of Rheumatology 2003;0(11):-
Objective To study the mRNA expression of HS1-associated protein X-1(Hax-1),an an- ti-apoptosis genc,in the peripheral blood mononuclear cells(PBMC)of patients with systemic lupus erythe- matosus(SLE),and further investigate the roles and significance of Hax-1 in the pathogenesis of SLE.Meth- ods Generation of longer cDNA fragments from serial analysis of gene expression(SAGE)tags for gene identi- fication(GLGI)was applied to identify the gene Hax-1 according to the Long SAGE tag.Then reverse tran- scription-polymerase chain reaction(RT-PCR)technique was used to semiquantitatively analyze mRNA ex- pressions of Hax-1 in PBMC from 34 active SLE patients and 25 healthy subjects.Results Compared with healthy controls,there was significant difference between SLE patients in the active stage and the normal controls(Z=-4.556,P<0.01).The average level of mRNA expression in active SLE group was higher than that in healthy controls.Significant difference was found between the group with mild SLE and either the moderate or the severe one(P<0.01).Conclusion The mRNA expression level of Hax-1 in active SLE group increase markedly,and to some extent,it is related to the activity of SLE.This provides a valuable basis for the further study on the role of apoptosis in SLE.
6.Expression of interleukin-13 receptor and its relationship to proliferation activity of human gliomas
Zhi-Bai XIA ; Xin-Jian WU ; Tie-Wei QI ; Hai-Jun WANG ; Zheng-Song HUANG ;
Chinese Journal of Microsurgery 2006;0(06):-
Objective To explore the gene expression of interleukin-13 receptor (IL-13R)?2 and its relationship to proliferation activity of human gliomas.Methods The gene expression of IL-13R?in 50 hu- man gliomas,2 malignant human glioma cell lines and 6 normal brain tissues were studied by RT-PCR.Ki-67 labeling index (Ki267 LI) of all sample were detecteded by immunohistochemical staining.Results Only one normal brain tissues expressed very low IL-13R?2 mRNA,whereas 35 (70%) of 50 human brain tumors expressed 1L-13R?2 mRNA.The positive rate and expression level of IL-13R?2 mRNA were increased with the ascending of WHO tumor grade.(former:rs=0.87;letter:rs=0.69,P<0.01).The difference of posi- tive rate and expression level of IL-13R 2?mRNA between the low grade and high grade tumors was statistical- ly significant,the proliferation activity of gliomas evaluated by Ki-67LI (Ki-67 Labeling Index,Ki-67LI) was positively correlated with IL-13R?2 gene expression and the tumor grade.Conclusion In human cerebral gliomas,IL-13R?2 genes may play an role in the malignant progression.The expression level of malignancy in molecular level and selecting the target of gene therapy.
7.Effects of controllable dynamic inhaled exposure of moxa smoke on LDL-r, ICAM-1 and morphology of heart tissue in rats.
Jia YANG ; Bai-Xiao ZHAO ; Li HAN ; Ping LIU ; Lei WANG ; Hua BAI ; Jian HUANG ; Jun-Tian LIU ; Chang HUANG ; Mao-Xiang ZHU ; Zhi-Hua YANG
Chinese Acupuncture & Moxibustion 2014;34(6):573-577
OBJECTIVETo observe the change of lipid metabolism and vascular endothelium as well as morphology of heart tissue in rats who were long-time exposed to moxa smoke with different concentrations in order to provide reference for safety assessment of moxa smoke on cardiovascular system.
METHODSOne hundred and sixty-eighty Wistar rats were randomly divided into a control group, a low-concentration group, a median-concentration group and a high-concentration group, 42 rats in each one. The rats were exposed to moxa smoke with concentration of 0%, 10%, 40% and 70%, respectively, for 20 min per day. After continuous intervention for six months, enzyme-linked immunosorbent assay (ELISA) was applied to measure the level of low density lipoprotein-receptor (LDL-r) and intercellular adhesion molecule-1 (ICAM-1) in blood serum in each group; the slices of heart tissue were stained with hematoxylin-eosin staining method to observe morphology change of heart tissue.
RESULTS(1) After the intervention of moxa smoke, the levels of LDL-r and ICAM-1 in the low-concentration group were not statistically different from those in the control group (both P > 0.05); the level of LDL-r in the median-concentration group was significantly increased, which was statistically different from that in the control group [(3.87 +/- 0.27) mg/mL vs (2.12 +/- 0.13) mg/mL, P < 0.01], however, the content of ICAM-1 was not obviously changed; although the level of LDL-r in the high-concentration group was presented with an escalating trend, it was not statistically different from that in the control group (P > 0.05) while the level of ICAM-1 was obviously increased (P < 0.01). (2) Under the light microscope, the abnormalities of cardiac muscle fibers and myocardial cell in each group were not been observed.
CONCLUSIONThe long-time intervention of low-concentration moxa smoke has no significant effects on lipid metabolism and vascular endothelium of rats, indicating that clinical application of low-concentration moxa smoke is relatively safe. The long-time intervention of moderate-concentration moxa smoke could significantly increase the clearance rate of cholesterol, implying the beneficial regulation of moxa smoke on lipid metabolism. The high-concentration moxa smoke could induce certain damage to vascular endothelium but its mechanism is in need of further research. The pathologic change of heart tissue could not be induced by moxa smoke with any concentration.
Animals ; Heart ; anatomy & histology ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipid Metabolism ; Male ; Moxibustion ; adverse effects ; Myocardium ; pathology ; Rats ; Rats, Wistar ; Receptors, LDL ; metabolism ; Smoke ; adverse effects ; analysis
8.Investigation of the layers and vascular density of the soft tissue in the inferior nasal portion.
Zhi-jun WANG ; Na WANG ; Hong-mei YANG ; Shu-ling BAI
Chinese Journal of Plastic Surgery 2007;23(1):65-68
OBJECTIVETo observe the layers and vascular structures of the soft tissue (including the skin) in the inferior part of external nose (nasal inferior portion) for providing the essential morphological data used for the correction of the bulbous nose and the tip of nose with thick skin.
METHODSUnder a light microscope, the density of microangium (vascular area/frame area) in each layer was measured with an image analyzer.
RESULTSThe stained soft tissue in the nasal inferior portion could be definitely divided into 5 layers: epidermis, dermis, superficial fascia, fibromuscular layer and perichondrium. According to the density of microangium, the sequence of these layers was: perichondrium, reticular layer, fibromuscular layer, sub-papillary layer, superficial fascia and papillary layer.
CONCLUSIONSThe soft tissue in the nasal inferior portion consists of 5 layers, which is similar to the structure of the face and neck. Both the total thickness of these 5 layers and the thickness of the dermis are much thicker than those of the Caucasus', and suit to rhinoplasty for bulbous nose According to the results of vascular density analysis, the skin thinning procedure in the rhinoplasty for bulbous nose should start from the superficial fascia firstly, then the fibromuscular layer.
Adult ; Humans ; Nose ; anatomy & histology ; blood supply
9.Safety of porcine fibrin sealant kit on ocular tissue following intravitreal injection
Bao-jie, HOU ; Jie, ZHAO ; Li, CHEN ; Wei-hong, XU ; Qing, XU ; Cui, HAN ; Fang, BAI ; Zhi-jun, WANG
Chinese Journal of Experimental Ophthalmology 2012;(11):1005-1008
Background Porcine fibrin sealant kit has been widely used in surgery for clotting and woundssealing.It is thought to be a substitution for inert gas and silicone oil in vitrectomy.But whether it produce toxic effect on retina or not after intravitreal injection is still studying.Objective This study aimed to investigate the retinal toxicity of porcine fibrin sealant kit following intraocular application.Methods Vitrectomy was performed on bilateral eyes of 15 pigmented rabbits.Porcine fibrin sealant kit of 0.5 ml was intravitreally injected in the lateral eyes of the rabbits as the experimental group,and equal volume of BSS was used at the same way in contralateral eye as control group.The inflammatory response of eye was observed under the slit lamp and ophthalmoscopy in 1 day,3,7,15,30 days after injection.Schi(o)tz tonometer was used to measure intraocular pressure(IOP) in 1 day,3,7 days,and retinal function was evaluated by electroretinogram (ERG) before operation and 30 days after operation.B-ultrasonic examination was carried out 30 days after surgery.Animals were sacrificed and eyeballs were obtained for retinal histopathological and ultrastructural examination in 30 days after operation.Results Complications appeared in 7 eyes of the experimental group,including cataract in 3 eyes,proliferation vitreoretinopathy and retinal detachment in 5 eyes,endophthalmitis in 1 eye,however,the cataract in 2 eyes and retina injury in 2 eyes were found in 15 control eyes.On the 30 days after injection,no inflammation was found in the 8 eyes without complication in the experimental group and 11 eyes in the control group.The IOP change was insignificant in different groups and various time points (Fgroup =0.008,P =0.929 ;Ftime =3.600 P =0.075).No significant difference was found in a-wave amplitude between groups and various time points(Fgroup =0.728,P =0.405 ; Ftime =0.516,P =0.482),and so was the b-wave amplitude (Fgroup =0.222,P =0.644 ; Ftime =0.057,P =0.814).On the 30th day after operation,arrangement of the retina was regular and no tissue edema and inflammatory cell infiltration were seen in both groups under the light microscope.Transmission electron microscope revealed that the ultrastructures of retinal cells,photoreceptor and retinal pigment cell were normal with the clear subcellular organelle,intact cellular membrane structure and mitochondria in both groups.Conclusions Domestic porcine fibrin sealant kit does not produce toxic effect on retina after intravitreal injection in rabbit.
10.General reproductive toxicity assessment in mice exposed to low-level ozone.
Zhi-jun ZHOU ; Zheng-shi ZHOU ; Bai-zheng TANG
Journal of Central South University(Medical Sciences) 2006;31(3):450-452
OBJECTIVE:
To explore the general reproductive toxicity in mice exposed to low-level ozone.
METHODS:
Low-level (0.09 approximately 0.18 mg/m3) ozone was created by 15 W ultraviolet light. The mice in 3 experimental groups and a control group were fed in low-level ozone environment or normal environment, respectively, and then the mating experiment was conducted. The pregnancy rate and the weight variations of the female mice were observed. The weight of the live fetuses was observed, and the appearance, bone and internal organs were checked for malformation.
RESULTS:
There were no significant differences in any indexes between the experimental groups and the control group.
CONCLUSION
Low-level ozone created by 15 W ultraviolet light may not have reproductive toxicity in mice.
Animals
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Dose-Response Relationship, Drug
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Female
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Fertility
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drug effects
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Inhalation Exposure
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adverse effects
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Male
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Mice
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No-Observed-Adverse-Effect Level
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Ozone
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toxicity
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Random Allocation
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Reproduction
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drug effects
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Ultraviolet Rays