1.Chitosan Promoting the Growth of in vitro Cultured Retinal Nerve Cells of SD Baby Rats
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2010;39(1):116-119
Objective To study the effect of chitosan on the growth of the in vitro cultured retinal nerve cells. Methods The retinal nerve cells of SD baby rats were cultured in vitro. The effects of chitosan at 0.065%,1.25%,2.5%,5%,10%,15% on the growth of the retinal nerve cells were measured by MTT method. The blank control group was created. Results 1.25%,2.5%,5%,and 10% chitosan could obviously promote the growth of retinal nerve cells,and 10% chitosan was the optimal concentration for the growth of nerve cells. Conclusion Chitosan can effectively promote the growth of retinal nerve cells,and it is necessary to undertake the further study of Chitosan.
3.Role of large conductance calcium-activated potassium channels in neuronal Ca2+ overload following traumatic brain injury
Hong ZHAO ; Wenhua YANG ; Xiangjun BAI
Chinese Journal of Trauma 2010;26(12):1140-1143
Objective To investigate the role of large conductance calcium-activated potassium channels (BK) in neuronal Ca2+ overload following traumatic brain injury (TBI). Methods Neuronal cells of C57BL/6 mouse cortex were collected and cultured. Patch-clamp technique was applied to investigate the changes of intracellular free calcium [Ca2+] i and firing frequency of neuronal action potentials in rest condition or evoked by 100 pA electric current lasting 500 ms after perfusion of Iberiotoxin ( 100 nmmol/L), a BK specific blocker. The cells were divided randomly into experimental group ( plus Iberiotoxin) and control group. Extracellular solution of cultured neurons was further perfused with KCl (20 mmol/L) to induce elevation of [Ca2 +]i and influence of Iberiotoxin ( 100 nmol/L) on amplitude of [Ca2 +] i elevation was determined. Results No significant changes of neuronal spontaneous action potential frequency and [Ca2 +]i were observed in rest condition after perfusion of Iberiotoxin (P>0.05).However, when evoked by electric current, the frequency of action potential was (10.4 ± 3.0) Hz,which was increased to ( 13.8 ± 3.7 ) Hz after perfusion of Iberiotoxin, with statistical difference (P<0.05 ). The [Ca2 +] i level was ( 14.21 ± 16.98 ) nmol/Lbefore perfusion with Iberiotoxin but was increased to (44.07 ± 34. 4) nmol/L after perfusion of Iberiotoxin (P < 0.05 ). Extracellular high concentration of KCl increased [Ca2 +] i of neurons, while perfudion of Iberiotoxin further elevated [Ca2 +]i (P < 0.05).Conclusion BK may play an important role in the regulation of neuronal [Ca2+] i and in neuronal Ca2+overload following TBI.
4.The cloning of mouse nicotinamide mononucleotide adenylyl-transferase gene and the detecting of its expression
Hong ZHAO ; Zichao YANG ; Jingyu ZHANG
Chinese Journal of Geriatrics 2011;30(10):866-868
Objective To construct eukaryotic expressing vector of the mouse NMNAT1 (nicotinamide mononucleotide adenylyl-transferase) gene and examine its ability to express the NMNAT1 gene in Hela cells.Methods The full-length NMNAT1 cDNA sequence was amplified by PCR and cloned into the plasmid of T-vector and then to pcDNA3.1 construct.The recombinant plasmid pcDNA3.1-NMNAT1 was identified by DNA sequencing and then transfected with Lipofectamine2000 into Hela cells.The expression of NMNAT1 was detected by real time quantitative PCR (qPCR) and Western blot after 48 h transfection.Results The recombinant eukaryotic vector carrying NMNAT1 gene was constructed successfully in a match with database and this vector could up-regulate the expression of the NMNAT1 gene both in mRNA and protein levels in Hela cells.Conclusions The eukaryotic vector carrying NMNAT1 gene (pcDNA3.1-NMNAT1) enhances the expression of NMNAT1 gene.
5.Antiviral Activity of the Effective Monomers from Folium Isatidis Against Influenza Virus in Vivo
Zhao LIU ; Zhanqiu YANG ; Hong XIAO
Virologica Sinica 2010;25(6):445-451
In order to evaluate the anti-influenza virus activity of the effective monomer from Folium Isatidis (FI) in vivo, we established mice model with viral pneumonia and divided them into 3 different dose groups, then observed their lung indexes, pulmonary pathological changes, pulmonary virus hemagglitination titers, living time and death rates. The results showed that the monomer could reduce the pulmonary index from 2.64 to 1.93, 1.63 and 1.40 (P<0.01) and decrease the hemagglitination titer from 1.15 to 0.84, 0.70 and 0.59 (P<0.01). In addition,the living time from 5.1d to 6.5d, 8.4d and 8.9d respectively(P<0.01). The high dose (75 mg/kg/d) has the similar effect with 100 mg/kg/d dose of virazole(P>0.05), and more effective than 200 mg/kg/d dose of antiviral liquor (P<0.05).
6.Professor T ian Cong-huo’s commonly used acupuncture methods and their clinical application
Tao YANG ; Rui WANG ; Hong ZHAO
Journal of Acupuncture and Tuina Science 2015;(2):74-82
To introduce Prof. T ian Cong-huo’s commonly used acupuncture methods and their clinical application, in order to summarize the famous doctor’s experience, promote the experience inheritance of Chinese acupuncture, and enhance the therapeutic effect of acupuncture.
7.Study on Intervention of Serum 3-deoxyglucosone byBu-Shen Hua-TanFang in Rats with Diabetes Mellitus and Cognitive Impairment
Hourui ZHAO ; Hong DAI ; Min YANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(10):2039-2043
This paper was aimed to study the influence ofBu-Shen Hua-TanFang (BSHTF) on the learning and memory function of diabetes mellitus (DM) rats. A total of 50 3-month-old male Wistar rats were randomlydivided into the control group, model group and 3 BSHTF groups (i.e., high-, middle-, and low-dose). A total of 35 mg·kg-1 streptozotocin (STZ) was intraperitoneally injected into the model and BSHT group. The same volume of citric acid-sodium citrate buffer solution was intraperitoneally injected into the control group. The intragastric administration of BSHTF at the dose of 20, 10, and 5 g·kg-1 was given to the BSHTF groups, respectively. The behavior and content of serum 3-deoxyglucosone (3-DG) were detected among DM rats after 30 days. The results showed that the levels of serum glucose in the model group and the BSHTF groups were obviously higher than that of the control group (P < 0.01). The swimming time to platform of rats in the model group was obviously prolonged compared to the control group with significant difference from the second day on (P < 0.01). The required time in different BSHTF intervention groups was obviously decreased compared with the model group from the 4th day (P< 0.05). The level of serum 3-DG in rats of the model group was obviously higher than that of the control group (P < 0.01). The level of serum 3-DG in rats of the middle-dose BSHTF group was obviously decreased compared with the model group (P < 0.05). It was concluded that BSHTF was able to improve the cognitive impairment among DM rats by reducing the levels of serum glucose and serum 3-DG.
8.Application of ProtEx~(TM):an exogenous FasL protein modification technology in the study of immunomodulation
Xiao GU ; Jin YANG ; Hong ZHAO
Chinese Journal of Organ Transplantation 2010;31(3):144-147
Objective By using ProtEx~(TM) technology to decorate target cells with an exogenous protein and observe its function for immunomodulation.Methods Donor splenocytes were decorated with chimeric SA-FasL protein.The expression of FasL and its impact on other molecules on the surface of splenocytes was detected.Heterotopic rat heart transplantation was performed from inbred WF rat to ACI rat.According to the different donor splenocytes perioperatively injected into recipients,the recipients were divided into 3 groups:SA-FasL group (n=23),SA group (n=20) and control group(n=10).Heart beating was examined by palpation on daily basis,the cessation was regarded as reiection,while surival over 100 days was considered as graft long-term survival.Diabetes models in C57BL/6 mice were induced by intravenous injection of STZ.Islets from BALB/C mice were decorated by SA-FasL protein,islets transplant was performed from BABL/c to C57BL/6 mice.Based on the different treatments of donor islets,recipients were divided into three groups:islet-FasL group (n=21),islet-SA group (n=21) and islet-untreated group(n=14).Rejection was identified while urine glucose was positive and blood glucose value was over 250 mg/ml for consecutive two days.Urine glucose negative over 100 days was regarded as graft long-term survival.Results The mean exDression rate of FasL in splenocytes was 94.49%±4.27% tested by flow cytometry and FasL showed red color 0n the surface of splenocytes by fluorescent stammg.FasL deeoration did not disturb the expression of CD3,CD4,D8,MHC I and CD80.The long-term survival rate o,f heart transplant showed significant difference between tWO groups(SA-FasL:70% vs SA:25%,P<0.05).67% FasL-decorated islets had normal function 30 days post transplant,and 29%FasL-decorated islets achieved long-term survival,which was significantly differed from the other groups(P<0.05).Conclusion ProtEx~(TM) is a simple,fast,safe and efficient approach for exogenous protein modmcation.Decoration of target cells with Chimeric FasL by this technique plays an umportant role in immunomodulatiorn.
9.Application of Sodium Hyaluronate in Cornea Preservation
Juan YANG ; Min ZHAO ; Hong LI
Journal of Chinese Physician 2001;0(03):-
Objective To study the effect of sodium hyaluronate(SH) on cornea preservation to provide a basis for eyes banks cornea preservation. Methods SH was applied in the short-term and middle-term cornea preservation. The preservation effect was evaluated by corneal morphology, trypan blue-alizarin red staining, activity assay of SDH and G-6-P enzymes, and ultrastructural observation of corneal endothelium. Results The above indices were obviously better in the experiment group than in the control group (P
10.Combined Toxicity of Volatile Organic Compounds and Ammonia in Indoor Air
Chaoyan OU ; Jinshun ZHAO ; Hong YANG
Journal of Environment and Health 1993;0(01):-
Objective To explore the combined toxicity of dominant volatile organic compounds(VOCs) and ammonia in indoor air of newly decorated rooms.Methods The gaseous mixture of several main components of VOCs and ammonia was prepared using their pure chemical reagents based on the percentage of mass proportion to each pollutant calculated by the medians of concentrations of formaldehyde,benzene,methylbenzene,dimethylbenzene,ethylbenzene and ammonia obtained from the in situ monitoring data on indoor air of 45 newly decorated apartments.The Kunming mice were exposed to the prepared gaseous mixture at doses of 3.93,8.47,18.24 and 39.35 g/m3 for acute toxicity test,and at doses of 3.935 g/m3(higher dose),1.970 g/m3(middle dose)and 0.393 g/m3(lower dose)for sub-acute toxicity test respectively.Results The LC50 of the prepared gaseous mixture was 26.84 g/m3 for Kunming mice.The sub-acute toxicity test showed that the lower levels of reticulocyte count of female mice in each dose group,the lower level of HCT of female mice dose group,the lower levels of PLT in serum of female and male mice in higher dose group,higher activity of ALT in serum of female mice in higher dose group showed significant differences compared with those of controls (P