2.Recording method of mouse pattern electroretinogram
Hai-xia, CHENG ; Song-tao, YUAN ; Qing-huai, LIU
Chinese Journal of Experimental Ophthalmology 2012;30(1):50-53
BackgroundMouse has been used in laboratory studies as the model of ocular diseases.Electroretinogram (ERG)is a non-invasive method for primary examination to evaluate retinal function.Though flash ERG has been widely applied in the mouse ocular disease model for the functional assessment of the retinal outer layer,pattern ERG(PERG) is seldom used for inner retinal evaluation.ObjeetiveThe present experimental study was to investigate the recording parameters and method,wave characteristics of PERG and influencing factors in mouse,and to build the foundation for further research.Methods Thirty C57BL/6 mice aged 6 weeks old were included in this research.RETLport ( Roland Consult,Germany) was adopted for the recording of PERG.The positive needle electrode was placed in the cornea,and the reference and earth electrodes were placed under the derma in the cheek and tail.The PERG under different temporal frequencies (0.5,1.0,2.0 and 4.0 Hz),and special frequencies (0.05,0.10,0.20 cpd) were recorded in a photopic environment,and different contrast ratio (95% and 99% ) of stimulator or different transmission bands ( 1-100 Hz,5-30 Hz) in the same temporal frequency and spatial frequency were regulated to analyze the influence on mouse PERG.The use of animals was in compliance to the Regulations for the Administration of Affairs Concerning Experimental Animals by the State Science and Technology Commission.ResultsThe latency of N1 PERG showed a negative N1wave at around 37 ms and positive P wave at about 86 ms in adult mice.The amplitude of N1-P was 2-6 μV.Different spatial frequency,temporal frequency and contrast can affect the final results,and the different temporal frequencies were statistically significant.The wave was stable and the amplitude was unaffected at 5-30 Hz transmission bands with pronounced interference (mean amplitude of N1-P waves were(3.40±0.71),(5.08±0.88),(3.21±1.54),(3.85±1.96)μV in 0.5,1.0,2.0,4.0 Hz,F=7.43,P=0.00).ConclusionsPERG wave from adult mouse is similar to that from human.It is a useful method in evaluating the inner retinal function.Appropriate stimulating parameters are critical for recording.
3.Expression of synapsin in visual cortex of developing mice
Qing, SU ; Song, HAN ; Ping, XI ; Junfa, LI ; Tao, FU
Chinese Journal of Experimental Ophthalmology 2014;32(5):388-391
Background The treatment timing and method of amblyopia rely on the plasticity of visual system.Synapsin is a family of presynaptic terminal specific protein.Its role in visual developmental plasticity is below understood.Objective To investigate the dynamic expressions of synapsin (T-synapsin),and phosphorylation of synapsin (p-synapsin Ⅰ a/b) in visual cortex of normal mice and further explore the role of synapsin in plasticity of visual system.Methods Forty-two clean neonatal C57BL/6 mice were collected.The mice were sacrificed at postnatal 7,14,21,28,35,42,60 days respectively to obtain the tissue samples of visual cortex.Expression levels of T-synapsin and p-synapsin in the visual cortex following the ageing were quantitatively detected using Western blot assay.Results The expression of synapsin in normal mice showed a dynamic increase with the ageing.The T-synapsin Ⅰ a/b/β-actin value in visual cortex was (21.32 ± 3.27) %,(56.27 ± 10.18) %,(77.05 ± 10.05) %,(83.75±10.52) %,(94.69±11.46)%,(98.75±5.86) % of adults mice (postnatal 60 days,P60) in the mice of postnatal 7,14,21,28,35,42 days,respectively,showing a significant difference among them (F =69.538,P < 0.001).Compared with the adult mice,the T-synapsin Ⅰ a/b/β-actin value in the mice of P7,P14,P21,P28 was significantly lower (all at P<0.05),but no significant difference was found between P35 and P60,P42 and P60 (P =0.280,0.798).The development trend of different synapsin subtypes,such as T-synapsin Ⅰ a/b,T-synapsin Ⅱ a,T-synapsin Ⅱ b and T-synapsin Ⅲ a,was not quite the same during the ageing.The expression of T-synapsin Ⅱ a and Ⅲ a increasing more slowly with development,and kept increasing until P60.Significant differences were found among various age of mice in T-synapsin Ⅱ a,Ⅱ b,Ⅲa respectively(F =42.492 55.595,39.172,all at P<0.001).The p-synapsin Ⅰ a/b level in the visual cortex elevated with the ageing of the mice,and that peaked in P21 mice,which was (2.86±0.17) times more than that in adult mice.After that,the expression level of p-synapsin Ⅰ a/b dropped rapidly.A significant difference was found in the p-synapsin Ⅰ a/b expression among different ages of mice (F =22.620,P < 0.001).Conclusions Synapsin level in visual cortex presents a developmental change which correlated with the onset and decline of the critical period.Synapsin is probably involved in the regulation of neural plasticity in visual cortex in critical period.
4.Impact of antihypertensive therapy compliance on risk of first-ever cerebral infarction among patients with essential hypertension
Xin XU ; Yingkui SONG ; Qing GENG ; Xiaoguang KONG ; Li TAO ; Xiaolei HU
Chinese Journal of General Practitioners 2009;8(5):305-308
ObjectiveTo evaluate the impact of compliance with antihypertensive therapy in pailents with essential hypertension on risk for their first-ever cerebral infarction.MethotisQuestionnaire survey and auxiliary examinations were conducted in 114 patients with essential hypertension hospitalized for acute cerebral infaretion at the First Hospihal Affiliated to Harbin Medical University during December 2006 to December 2007,as well as in another 114 patients with essential hypertensive without history of cerebral infarction as controls during the sanle period.Univariate and multivariate 10gistic regression analyses were performed to study the relationship between first-ever cerebral infarction and compliance with antihypertensive agents and other relevant factors.ResultsAntihypertensive agents compliance,course of hypertension,and history of smoking and alcohol drinking could significantly affect their first occurrence of cerebral infarction in patients with essential hypertension(P<0.05),with odds ratios(OR)of 0.429(95%C10.186-0.993) and 2.142(95% CI 1.052-4.364)for good and poor compliance with antihypertensive agents,respectively,as compared to those without antihypertensive treatmenL Mild drinking was a protective factor for cerebral infarction with an OR of 0.494(95%CI 0.252-0.968).kngth of hypertension with 10-19-years and more than or equal to 20 years.as compared to those with le88 than five years of hypertension,was also a risk factor for it,with an OR of 2.118(95% CI 1.075-4.174).ConclusionsCompliance of essential hypertensive patients with antihypertensive therapy was an important factor that affect their contracting first-ever cerebral infarction.Good compliance could obviously refrain them from it.Patients with poor-compliance or without treatment prone to contract cerebral infarction more easily than those with good compliance.It is necessary to improve compliance with antihypertensive agents in patients with essential hypertension as soon as possible.as well as quitting smoking and limiting alcohol drinking for prevention and control for their first-ever cerebral infarction.
5.Determination of content and encapsulation efficiency of leuprolideacetate microspheres
Hui TAO ; Bo WANG ; Song ZENG ; Limei HE ; Qing HUANG ; Huaqing LIN ; Rong LIU
Chinese Journal of Biochemical Pharmaceutics 2017;37(1):60-62
Objective To establish a determination method for leuprolide acetate microspheres. Methods HPLC was performed on Inertsil ODS-SP (150×4.6 mm×5μm) with mobile phase consist of 0.1 mol/L of Ammonium Dihydrogen Phosphate(adjust its pH to 7.00±0.05 with Ammonium Hydroxide) and Acetonitrile in ratio of 3:1(V/V),and the flow rate was 1.0 mL/min.The wavelength was 220 nm and column temperature was 30℃. The injection volume was 20 μL. Results The linear range of leuprolide acetate was 20.0-160.0μg/mL (r=0.9999) with an average recovery of 99.80%,RSD=0.63%(n=9). Conclusion The method of HPLC was accurate,reliable and specific, which could be used to determinate the assay of leuprolide acetate microspheres and for quality control of microspheres.
6.Inhibition effect of c-Met inhibitor on proliferation of lens epithelial cells
Xiao-Ling, AN ; Wei-Ming, WANG ; Qing, HAN ; Jin-Hua, TAO ; Jin-Song, ZHANG
International Eye Science 2006;6(4):743-744
AIM: To observe the inhibition of c-Met inhibitor on proliferation of lens epithelial cells (LECs).METHODS: Human's LECs were cultured and hepatocyte growth factor (HGF) and K252a were added to second passage of cells supplied with Dulbecco's modified eagle's medium (DMEM). MTT assay was used to examine the proliferation of LECs, and Western-blot was used to detect the expression change of Bcl-2 and Caspase-3.RESULTS: The photodensity (A) of HGF (50nmol/L) + K252a (30nmol/L) was not significantly different from that of DMEM control (P>0.05). The expression of Bcl-2 and Caspase-3 were not significantly different from that in the control group.CONCLUSION: K252a, the inhibitor of c-Met, can effectively inhibit the proliferation of LECs.
7.Effects of serum enatninine Gumibao (Chinese character: see text) on the aroliferation and differentiation of osteoblast induced by dexamethasone.
Yi SONG ; Hong JIAN ; Dao-fang DING ; Ling-hui LI ; Guo-qing DU ; Jin-tao LIU ; Hong-sheng ZHAN
China Journal of Orthopaedics and Traumatology 2014;27(8):668-672
OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.
METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.
RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.
CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.
Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley
8.The effects of immunity induced by BCG priming and tuberculosis fusion protein AMM vaccine boosting
Wenwen JIANG ; Tao JING ; Hongjuan YU ; Qing LI ; Juan YI ; Yu LUO ; Nannan SONG ; Ying ZHANG ; Bingdong ZHU
Chinese Journal of Microbiology and Immunology 2009;29(7):631-635
Objective To investigate the boosting efficiency of a subunit vaccine consisting of the fusion protein Ag85B-Mpt64190-198-Mth8.4 (AMM) , dimethyl-dioctyldecyl ammonium bromide (DDA) and BCG polysaceharide nucleic acid (BCG-PSN) on the primed inoculation with BCG. Methods The AMM subunit vaccine was composed of fusion protein AMM, adjuvant DDA and BCG-PSN. The first mouse experi-mental group was immunized with BCG first, then boosted with the AMM subanit vaccine in the 10th week. The second experimental group was boosted with the AMM subunit vaccine in the 8th week and the 10th week respectively with a two weeks interval after the primed with BCG. Two control groups were treated re-spectively with physiological saline alone and BCG alone. After the primed inoculation, ELISPOT and ELISA were used for the detection of the cell-mediated and humoral immune response in week 14 and week 22 re-spectively. Furthermore, the immunized mice were challenged with live BCG to mimic tuberculosis infection in the 22nd week after the primed inoculation. Subsequently the T cell typing and humoral response were de-tected by flow cytometry and ELISA, respectively. Results ( 1 ) The level of secreting IFN-γ: 14 weeks af-ter the primed inoculation,with the stimulation of the specific antigen-Ag85B, the number of cells secreting IFN-γ in the second experimental group (135±14) was more than BCG alone immunized group (19±16), t = 10. 98, P < 0.01. In the 22nd week, the number of cells secreting IFN-γ in the second experimental group (208±11) was still more than BCG alone group (57±18), t =6.43, P <0.01. (2) The level of humoral immune response: the IgG1 antibody titer in the second experimental group was obviously higher than that in the first experimental group. However, the ratio of IgG2a to IgG1, as the index reflecting the Thl-type immune response, in the experimental group 2 was lower than that in the experimental group 1. (3) The contents of CD4+ CD25+ T cells after challenged with live BCG strain: the first and the second ex-perimental groups were both higher than the BCG alone group (t1 = 3.08, t2 = 3.16, P < 0.05 ). Conclu-sion Boosting the BCG-pfimed mice with tuberculosis AMM subunit vaccine twice can induce higher level of cell-mediated and humoral immune response than BCG alone, which could activate the regulative immune response at the same time.
9.Influence of Velocity of Flow of Contrast Media on Spiral CT Portography
Jianruing LEI ; Peng DONG ; Guanmin QUAN ; Tao YUAN ; Qing LIU ; Changqiang QU ; Jingying BU ; Peng SONG ; Wujie LIU
Journal of Practical Radiology 2010;26(2):258-261
Objective To investigate the effect of different injection rate of contrast medium on the quality of multislice spiral CT portography(SCTP). Methods Thirty patients with no prominent difference in the age and weight randomly divided into 3 groups (10 cases each group) underwent contrast-enhanced CT scan of abdomen. Non-ionic contrast medium (300 mgⅠ/ml, 1.5 ml/kg) was injected through antebrachium veins with power injector at the rates of 2.0 ml/s,3.0 ml/s and 4.0ml/s. Arterial phase acquisi-tion was made using the software of automated bolus triggering with a ROI placed on abdominal aorta when 100 HU was reached, and 7 seconds and 20 seconds later, portal phases were done respectively. The attenuations of portal vein(PV) and hepatic parenchy-ma of 3 groups were measured on source images. Then the oblique axial and coronal maximum intensity projection(MIP) maps were reconstructed at workstation and PVs were observed. Statistics analysis was made with software of SPSS 11.5. Results The attenu-ations of PV in 2.0ml/s, 3.0 ml/s and 4.0 ml/s groups were (150.80±21.16)HU, (170.90±17.26)HU and [181.90±22.88) HU respectively. There was obvious difference between 2.0ml/s and 4.0ml/s groups for atteuation of PV(P=0.017). The differences of CT attenuation of PV-hepatic parenchyma in three groups were (50.20±17.40) HU, (67.10±23.08) HU and (76.20±22.75) HU respectively. Prominent difference was also found between 2.0 ml/s and 4.0 ml/s groups(P=0.039). The grades of segment of PV displayed on SCTP maps were 4.20±1.14,4.90±0.99 and 5.50±0.53 in 2.0 ml/s,3.0 ml/s and 4.0 ml/s groups respectively, which were of obvious differences between them (P=0.013). Conclusion Injection rate of contrast medium dose influence the quality of SCTP of PV,SCTP quality is best with velocity of flow at 4.0 ml/s.
10.Protective effect ofα-mangostin on retinal light damage in mice
Yuan, FANG ; Tu, SU ; Ping, XIE ; Song-Tao, YUAN ; Wen, FAN ; Yi-Dan, XU ; Zi-Zhong, HU ; Qing-Huai, LIU
International Eye Science 2015;(7):1143-1147
AlM:To discuss the protective effect ofα-mangostin on retinal light damage in mice.METHODS:Totally 30 Balb/c mice, aged 6~8wk, were randomly divided into the control group, light-exposure group and α-mangostin group. Every group contained 10 mice. Mice of α-mangostin group were treated with alpha-mangostin at the dose of 30mg/( kg · d ) body weight by intragastric administration daily for 7d, and then exposed to white light at the 5th d. The light-exposure group and α-mangostin group were exposed to 5 000 ± 200lx white light-emmiting diodes (LEDs) for continuously 1h to establish the mice model of retinal light damage. Flash -electroretinograme was recorded 72h after light exposure. The changes in retinal morphology of mice were observed by light microscopy. Retinas were extracted to detect the malondialdhyde ( MDA ) content change of the retinal homogenate.RESULTS: Flash-electroretinogram ( F-ERG ) showed that retinal dysfunction was less severe in α-mangostin group than in light-exposure group ( P<0. 05 ). Light microscopy test showed that retina structural damage was less severe in α-mangostin group than in light-exposure group (P<0. 05). The level of MDA in retinal tissue of α-mangostin group was significantly lower when compared with light-exposure group (P<0. 05).CONCLUSlON: α-mangostin inhibits lipid peroxidation induced by light damage and protect retina against light damage.