1.The method of monitoring sleep state and respiratory rhythm in freely moving rats.
Jing WANG ; Guang-fa WANG ; Yi-ling LING
Chinese Journal of Applied Physiology 2004;20(3):310-312
AIMTo devise a new method for testing the animal model of sleep apnea syndrome (SAS).
METHODSWe simultaneously monitoring the respiratory wave through a micro-pressure transducer and the electrical signals of both electrocorticogram (ECoG) and electromyogram (EMG), for detecting the sleep-wake states and the respiratory rhythm in rats. The recording data are analyzed by the manual-computer work.
RESULTSThe ECoG and EMG signals are stable and easily to distinguish. We can accurately detect the sleep-wake state and the sleep apnea events in rats by these signals.
CONCLUSIONThe method is reliable to test the small animal model of SAS, and is easily to operate.
Animals ; Circadian Rhythm ; physiology ; Electroencephalography ; methods ; Monitoring, Physiologic ; methods ; Rats ; Rats, Sprague-Dawley ; Respiration ; Respiratory Physiological Phenomena ; Sleep ; physiology
2.Determination of antigenic properties of VirB9, a protein of type Ⅳ secretion system of Brucella
Jin-ming, FAN ; Fa-xing, WANG ; Bo, ZHANG ; Ling, JIANG ; Bei, LI
Chinese Journal of Endemiology 2013;(3):263-266
Objective To detect the immunogenicity of VirB9,a protein of type Ⅳ secretion system of Brucella.Methods Full length VirB9 gene was cloned into plasmid pET32a and expressed in Escherichia (E.) coli BL21 (DE3).Expression of recombinant protein was induced by isopropyl beta-D-thiogalactopyranoside (IPTG) and the recombinant fusion protein was purified by affinity chromatography on Ni2+-conjugated chelateing sepharose.The purity of the purified protein was ascertained by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDSPAGE) and the concentration was measured by bicinchoninic acid (BCA) protein assay kit.Animal model was established by immunizing BAL B/c mice with live vaccine strain S19 of Brucella and the mice immunized with phosphate buffered saline (PBS) as control.The blood of immunized mice was acquired after 4 weeks.Antibody against VirB9 in S19 immunized mice was detected by Rose Bengal plate agglutination test and serum tube agglutination test; IgG antibody titers against VirB9 in immunized mice were determined by enzyme linked immunosorbent assay(ELISA).At the 35th day,the immunized mice and control mice were killed and spleens were collected.The splenocytes were harvested and stimulated with each of VirB9,concanvalin A(ConA) or medium in triplicate.Production of gamma interferon (IFN-γ) was determined by enzyme-linked immunospot assay (Elispot).Results The full length of VirB9 gene was cloned into pET32a.The recombinant VirB9 protein was expressed at 43 × 103 in relative molecular mass and the purity of the purified recombinant VirB9 protein was above 97% in SDS-PAGE and the concentration was 1.6 g/L in BCA protein assay.The antibody of VirB9 was detected in all S19 immunized mice but not PBS immunized mice by Rose Bengal plate agglutination test.The antibody titer in all S19 immunized mice was > 1 ∶ 800 or > 1 ∶ 3200 by tube agglutination test and ELISA,respectively.Meanwhile,the protein stimulated stronger IFN-γresponse in immunized mice than that in the control mice(147 cells Vs 38 cells).Conclusion VirB9 can stimulate humoral and cellular immunity and it might be an appropriate target for developing subunit vaccine against Brucella.
3.Experimental study on co-culture of salivary adenoid cystic carcinoma cells and ganglia.
Ling GU ; Rong-fa BU ; Dong-sheng WANG ; Ling-ling E ; Guo-xiong ZHU
Chinese Journal of Stomatology 2012;47(1):48-51
OBJECTIVETo construct the co-culture models of salivarya denoid cystic carcinoma (SACC) cells and dorsal root ganglia (DRG) of chickens and investigate the promotive effects of SACC on neural tissue.
METHODSGlass-base culture dish was adopted to construct co-culture model of SACC-83 cells and DRG. SACC-83 cells were seeded in the medium pore with DRG around them. Outgrowth of neuronal processes was observed. Then DRG was cultured in the conditioned medium of SACC-83, with the groups of conditioned medium of MC3T3-E1 and HGF, the group of cell lysis buffer, the groups of serum-free medium and serum-plus medium as the controls. Outgrowth of neuronal processes was also recorded and compared with control groups.
RESULTSIn the co-culture model of tumor and neuronal tissue, SACC-83 cells produced a suitable microenvironment in which neuronal processes remarkably grow. Neuronal processes of most DRG displayed growth tendency toward SACC. The group of conditioned medium from SACC-83 manifested obvious promotive effects on DRG.
CONCLUSIONSCo-culture model of tumor and neuronal tissue was successfully constructed, with which the promotive effects of tumor on outgrowth of neuronal processes could be observed. So hypothesized that SACC could secrete some neurotrophic factors to guide peripheral nerves gemmating and to trigger the cascade of the neural invasion in succession.
Animals ; Carcinoma, Adenoid Cystic ; pathology ; Cell Line ; Cell Line, Tumor ; Chickens ; Coculture Techniques ; Culture Media ; Ganglia, Spinal ; growth & development ; Gingiva ; cytology ; Humans ; Osteoblasts ; cytology ; Salivary Gland Neoplasms ; pathology
4.Clinical application of portable echocardiography system in acute paroxysmal dyspnea
Lin CHE ; Ming-Zhong ZHAO ; Ming LUO ; Jin-Fa JIANG ; Bing DENG ; Yuan-Ling LI ; Le-Min WANG ;
Chinese Journal of General Practitioners 2003;0(03):-
Objective To study the clinical value of portable echocardiography system in diagnosis for acute paroxysmal dyspnea.Methods Clinical data of 81 patients with acute paroxysmal dyspnea recorded by a portable echocardiography apparatus at their bedside were retrospectively analyzed,and compared to those of 45 patients by conventional echocardiography.Results The 2D images in portable echocardiograph were similar to those of conventional echocardiograph.Diagnosis could be established in 74 (91.4%),corrected in six (7.4%) and not confirmed only in one (1.2%) of 81 patients with acute paroxysmal dyspnea by portable echocardiography system.And,portable echocardiography system could be used to diagnose pericardial effusion and to monitor perieardial puncturing and draining at bedside. Conclusions Portable echocardiography systems can provide rapid,accurate and valuable information on diagnosis and treatment for acute paroxysmal dyspnea,and make its clinical intervention accurate,scientific and effective,bringing echocardiography performed at bedside possible.
5.Differentially expressed genes associated with cold acclimation.
Fa-Qing YANG ; Ling-Jia QIAN ; Wan-Yin WANG ; Hui-Rong REN ; Da XU
Acta Physiologica Sinica 2003;55(3):360-363
To investigate the upregulated genes associated with cold acclimation, a cold acclimation model was established based on Balb/C mouse. mRNA of muscle and liver were isolated, and the upregulated genes of these tissues were studied by representational differential analysis (RDA). The upregulated genes then were sequenced and searched by Blast software in GenBank database. The results showed that some genes were upregulated and possibly associated with cold acclimation. Three of these genes, transferrin, fibrinogen B-beta-chains and a new gene fragment (Genbank ID: AF454762), were confirmed to be upregulated by RNA slot-blot analysis. The finding of these genes might contribute to further understanding of the molecular mechanisms of cold acclimation.
Acclimatization
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genetics
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Animals
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Cold Temperature
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Gene Expression
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Liver
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metabolism
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Male
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Mice
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Mice, Inbred BALB C
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Muscle, Skeletal
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metabolism
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Transcriptome
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Up-Regulation
6.Application of flowing injection of hydride generation atomic fluorometric method to assay blood selenium content
Cheng-bo, CHE ; Jun-rui, PEI ; Shao-chen, LI ; Ming-fa, LIU ; Li, SONG ; Yu, WANG ; Yang, LIU ; Ling-wang, ZHOU ; Tong, WANG
Chinese Journal of Endemiology 2008;27(5):555-557
Objective Using flowing injection hydride generation atomic fluotometric method(FI-HG- AFM)to detect the whole blood selenium content.Methods A series of standard selenium(1,2,4,6,8.10μg/L) were dectecte by FI-HG-AFM,and the precision and accuracy of the method were observed.The selenium contents of 89 blood samples were detected by FI一HG-AFM and 2,3-diaminonaphthalene fluorometric method(DFM) respectively,and the correlation of two methods Was analyzed.Results The detection limit and recovery rate of FI-HG-AFM were 0.138μg/L and 96.4%,respectively.The relative standard deviation(RSD)was maximized at 1 μg/L(4.406%).The RSD diminished along with the increase of the concentration,and it declined to 0.512%at 10 μg/L.There wag no significant difference(t=1.7878,P>0.05)between the result8 of FI-HG-AFM[(45.9± 14.5)μg/L]and DFM[(47.5 ±13.3)μg/L],and the two methods were in highly positive correlation(r=0.8143,P< 0.01).Conclusions The whole blood selenium content call be rapidly and exactly detected by FI-HG-AFM,and the method are highly consistent with DFM.
7.Clinical application of MR diffusion tensor imaging in lumbar disc annulus fibrosus
Hai-Tao YANG ; Ren-Fa WANG ; Juan WANG ; Xiao-Ling GAO ; Feng LI ; Hai-Dong ZHANG ; Li-Ming XIA ; Cheng-Yuan WANG ;
Chinese Journal of Radiology 1999;0(10):-
0.05).Seven cases in LDH group and 9 cases in HID group were found in FT picture.The mean DCavg value in annulus fibrosus disruption was significantly larger (1.01?0.10)?10~(-9)mm~2/s and the mean FA value(0.15?0.03)was significantly smaller than those in normal place(P
8.Expression of 8-hydroxy-2-deoxy guanosine, thioredoxin reductase 1 and glutathione peroxidase 1 in myocardium of autopsy patients with Keshan disease
Jun-rui, PEI ; Ming-fa, LIU ; Yang, LIU ; Hong-qi, FENG ; Zhi-yi, ZHANG ; Ling-wang, ZHOU ; Xue-kuan, ZHONG ; Tong, WANG
Chinese Journal of Endemiology 2012;31(6):631-634
Objective In this study,we investigated the relationship between oxidative stress,selenoproteins level and onset of Keshan disease (KD) through detecting the expression of 8-hydroxy-2-deoxy guanosine (8-OH-dG),thioredoxin reductase 1 (TrxR1) and glutathione peroxidase 1 (GPx1) in myocardial tissue.Methods Myocardium samples of autopsy patients including 8 cases of KD (KD group included 4 acute KD and 4 chronic KD) and 9 cases of non-KD (control group) were immunohistochemically stained for 8-OH-dG,TrxR1 and GPx1.The staining intensities subsequently quantified by using Olympus Image-Pro Plus 6.0 software.Results The positive rate of 8-OH-dG expression in myocardial nuclei was higher in the case group[(68.6 ± 20.4)%] than that of the control group[(2.4 ± 1.5)%,t =8.515,P < 0.05].In addition,the positive rate of 8-OH-dG expression in acute KD[(91.7 ± 3.7)%] was significantly higher than that of chronic KD[(53.2 ± 7.9)%,t =6.409,P<0.05].The distribution of TrxR1 and GPx1 was not associated with the distribution of myocardial damage.The expression of these two selenoproteins in KD group (401340 ± 59865,497590 ± 197082) were both lower than that of control group(2790300 ± 379298,1348400 ±615840; t =-28.493,-6.016,respectively,all P<0.01).Conclusions Oxidative damage is detected in myocardium tissue of KD,and 8-OH-dG expression is associated with the degree of myocardial damage in KD.Selenoproteins,TrxR1 and GPx1,may be closely related to the pathogenesis of KD.
9.Experimental study on cardiac pathological change in rats fed with corn and bean puree of Keshan disease area
Li-jun, ZHANG ; Ming-fa, LIU ; Jie, CHEN ; Shao-chen, LI ; Jun-rui, PEI ; Ling-wang, ZHOU ; Yang, LIU ; Tong, WANG ; Wei-han, YU ; Bao-xiong, TI
Chinese Journal of Endemiology 2009;28(3):291-293
Objective To investigate the myocardial damage in rats fed with corn from Keshan disease area added with bean puree. Methods Male Wistar rats were randomly divided into 3 groups according to their body weights, and fed with corn, corn from Keshan disease area added with bean puree, corn from non-endemic area. The GSH-Px activity of vena cardalis blood was examined in 1 and 3 months, rats were sacrificed after being fed for 6 months to examine the heart changes with HE stain. Results The three groups of GSH-Px activity were different in 1 and 3 months respectively(F=23.60,72.46, all P<0.01); GSH-Px activity was (181.58±22.15), (44.76±28.59)U/L in rats fed with corn, was (195.03±17.66), (30.38±3.35)U/L in those fed with corn added with bean puree from Keshan disease area, lower than the group fed with corn of non-endemic area[(340.90±95.42), (125.17±13.64)U/L, all P < 0.01]. But the difference of GSH-Px activity between simple corn group and corn adding bean puree groups of Keshan disease area was not obvious(P>0.05). Myocardial damage incidence of the three groups was 3/9,1/9,2/7. Difference among three groups did not have statistical significance(χ2=1.33, P> 0.05). Conclusions Only corn from Keshan disease area may induce myocardial damage pathology change. Adding bean puree into corn does not increase damage.
10.Rescue of postoperative carotid blowout in head and neck neoplasms
Ming-Bo LIU ; Wen-Wing WU ; Jia-Ling WANG ; Jun WANG ; Liang-Fa LIU ; Hui ZHAO ; De-Liang HUANG ; Dong-Yi HAN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2010;45(5):410-413
Objective To investigate the prevention and rescuing measures of postoperative fatal bleeding induced by carotid blowout in head and neck tumors. Methods Seven cases with postoperative carotid bleeding treated from October 2003 to August 2009 were reviewed retrospectively. Of the patients, 6 were with common carotid blowout and one with internal carotid artery blowout. All patients underwent preor post-operative radiotherapy for primary head and neck tumours and 3 patients had neck defect repair with deltopectoral skin flap, frontal flap or free radial arm flap respectively. After carotid blowout bleeding, the patients were treated in time with X ray transcatheter intervention including transcatheter arterial embolization (TAE) and self-expanding covered stent implantation, followed by repairing the carotid region with appropriate myocutenous flaps. Results Of 7 patients with carotid blowout, 5 patients were successfully rescured with X ray transcatheter intervention, of them 2 with self-expanding covered stent implantation and 2 with TAE respectively, and other 2 patients died due to rapid bleeding. Of the successfully rescued patients, 2 patients were with the repair of carotid area by pectoralis major myocutaneous flap, one by submental flap and one by local flap, but another one not with flap repair. Follow-up showed the 3 patients rescued with self-expanding covered stent implantation were survival for 6,12,and 20 months, respectively, and the 2 patients rescued with TAE died of repeated carotid blowout in 2 and 13 months later, respectively. Conclusions The planned and timely X ray transcatheter intervention is an effective method to treat carotid blowout bleeding in the patients underwent head and neck tumour surgeries. Compared with TAE, selfexpanding covered stent implantation may be more reliable for restoring the blood supply of head and neck region, with less complications. One-stage repair of caratid region with myocutenous flap is of great importance to protect the carotid and to promote the wound healing.