1.The influence of salvianolic acid B to blood brain-barrier with oxygen/glucose deprivation and reintroduction in vitro
Jihong HE ; Haiyang YANG ; Jiang LONG
Chongqing Medicine 2015;(33):4611-4614
Objective To explore the influence of salvianolic acid B(SalB) to blood brain‐barrier(BBB) in vitro oxygen/glu‐cose deprivation and reintroduction .Methods We used the human brain microvascular endothelial cells and C6 cells to build BBB model that in vitro ,and the models was divided into normal groups ,experiment groups ,treatment groups .The Earle′s liquid and the condition of 93% N2 ,2% O2 ,5% CO2 replaced the normal culture environment to simulate environment of oxygen /glucose depriva‐tion and reintroduction .The changes of BBB permeability was detected by the horseradish peroxidase permeability test .The tight junctions(TJs) was detected by immunofluorescence and electron microscope .Results The permeability of treatment group was significantly lower than the experiment group (P < 0 .05) ;The TJs of normal group was continuous and density ;the experiment group′s TJs was interruption and osteoporosis ;and the observation of treatment group′s TJs was between of the two group above . Conclusion The SalB can directly protect the BBB during oxygen /glucose deprivation and reintroduction ,and decrease the permea‐bility .The investigation also provided the research foundation for the SalB to protect the BBB .
2.Expression changes of Tau protein in retinal ganglion cells and oligodendrocytes following axonal injury in rats
Zhaohui HE ; Xiaochuan SUN ; Feng LI ; Long JIANG ; Luping ZHENG
Chinese Journal of Trauma 2009;25(7):587-591
Objective To investigate expression changes of Tau protein in retinal ganglion cells (RGCs) and oligodendrocytes (Ols) after stretch injury in rats and explore the relationship of Tau protein with pathological changes after axonal injury. Methods Morphological changes of optic nerves, RGCs and OLs after stretch injury were examined under light microscope in control group, stretch only group, heat stress only group and heat stress pretreatment plus stretch group. The expressions of Tau protein in RGCs and OLs after heat stress and/or stretch injury were observed by using immunohistechemical stai-ning. Results Pathological changes of axons, RGCs and OLs were identified morphologically or quan-titatively after stretch injury to the optic nerves, which was significantly ameliorated through pretreatment with heat stress plus stretch injury. The expressions of Tau protein in RGCs and OLs were increased in stretch only group. There was no significant expression change of Tau protein in heat stress only group. Expression of Tan protein was obviously decreased in heat stress pretreatment plus stretch group. Con-clusions Both neurons and glial cells are involved in pathological process after axonal injury. The ex-pression changes of Tau protein are probably related to delayed axotomy and neuron apoptosis. Heat stress can relieve the impairment of cystoskeleton through decreasing and delaying the expression of Tau protein.
3.The effect of valproic acid on behavior and senile plaques in the male and female APP/PS1 double transgenic mouse model
Zhimin LONG ; Lei ZHAO ; Rong JIANG ; Guiqiong HE
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(3):205-207
Objective To investigate whether valproic acid (VPA) affect spatial learning memory and senile plaques in the APP/PS1 double transgenic AD mouse model of different gender. MethodsTwenty 3-month old APP/PS1 double transgenic AD mice,male and female mouse evenly,were randomly divided into VPA-treated and saline-treated groups ( 10 for each group). 30 mg· kg-1 · d-1 of VPA and the same amount of saline were peritoneally injected into mice for 4 weeks. Morris water maze was conducted to check the effect of VPA on the capability of spatial learning and memory of AD mouse model. Immunohistochemical staining was used to examine the effect of VPA on the morphological changes in the brains of mice. ResultsVisible platform test showed that VPA-treated and saline-treated mice had similar escape latency (P>0.05) and path length (P>0.05) ,the swimming speed between male and female mice had no difference (P>0.05). Hidden platform test showed that VPA treated mice had a significantly shorter latency (P<0.01) and path length (P<0.01) to reach the platform compared with saline-treated mice. Meanwhile, both in VPA-treated and control groups, the male mice had a shorter correlation escape latency and path-length than female mice had(P<0.05 ). Immuohistochemical staining showed that the number (11.23±3.78) of senile plaques (SP) in the cerebral cortex and hippocampus of VPA-treated male mice were notably decreased than that(28.17 ±3.46) in the control group ( t= 14.67, P<0.01 ),furthermore,the number of SP in the cerebral cortex and hippocampus of VPA-treated male mice was significantly reduced,as compared with which (20.36 ±4.21)in the VPA-treated female mice(P<0.05). ConclusionVPA can significantly lower formation of SP, and remarkably improve the capability of spatial learning and memory of APP/PS1 transgenic mice,which have gender difference.
4.Changes of Intracerebral Amino Acid Transmitters in Periventricular Leukomalacia of Newborn Rats Observed with Microdicalysis Technology
long, ZHANG ; li, JIANG ; ting, WANG ; nong-yue, HE
Journal of Applied Clinical Pediatrics 2006;0(20):-
Objective To observe the changes of intracerebral amino acid transmitters in the periventricular leukomalacia (PVL) of newborn rats with microdialysis and so as to explore the role of excitotoxicity in PVL.Methods Replicated the model for PVL at the age of postnatal day 5 (P5) by intracerebral injection of 3-nitropropionic acid (3-NPA).Before injection of 3-NPA,and 15 min,30 min,45 min,60 min,75 min,90 min after injection of 3-NPA,collected the sample of extracellular fluid (ECF) at the corpus callosum above the left ventricle through microdialysis,respectively.After microdialysis,the experimental rats were allowed to survive to P6-P14,and then they were killed and the brains were prepared for HE stain.The amino accids of dialysate were quantified through high performance liquid chromatography(HPLC),and then the excitotoxic index (EI) was calculated.Results Fifteen min to 45 min after injection of 3-NPA,the concentrations of glumate (Glu) and aspartic acid (Asp) of ECF elevated significantly,and then returned to the normal levels.Fifteen min to 75 min and 15 min to 30 min after injection of 3-NPA,the concentrations of glycine (Gly) and GABA significantly elevated,respectively,and returned to normal levels at 90 min and 45 min after injection of 3-NPA,respectively.But the EI,which indicated the balance of excitatory amino acids(EAAs) and inhibitory amno aciols(IAAs),significantly elevated 15 min to 75 min after injection of 3-NPA,then retured to normal level after 90 min.Sub-cortical and periventricular white matter rarefaction and significant lateral ventricle enlargement were observed in HE staining.Conclusions Changes of intracerebral amino acid transmitters in the PVL of newborn rats show regularity:EAAs,IAAs of ECF and EI elavate in the early stage,and then return to the normal level quickly.It indicates that excitotoxicity play a great role in PVL,especially at the early stage.Therefore,the preventions of PVL must be executed at the early stage.
5.Toxicity of sodium orthovanadate on guinea pig myocardial cells
Jiang LONG ; Huaguang WANG ; Shuzhuang HE ; Baofeng YANG ;
Chinese Pharmacological Bulletin 1986;0(04):-
AIM To study toxicity and mechanism of SOV on ventricular myocardial cells. METHODS Cyclical perfusion was used to acutely detach the mature guinea pig myocytes into single cells. The CK,LDH activity in a medium of myocardial cells was measured for 30,60,120 and 180 min after the effect of SOV at 1,10, 100 ?mol?L -1 and 1 mmol?L -1 . Rates of cell viability and apoptosis were also measured. The protein content and Na +,K + ATP enzyme activity of the myocardial cells were measured as well. An electron microscope was used to investigate cell nucleus and organelle morphology. RESULTS The release of CK, LDH and protein from the myocardial cell gradually increased after administering SOV at 100 ?mol?L -1 and 1 mmol?L -1 to the cells. While ,the cell activity decreased. CK release, which indicated the extent of myocardial cell injury, was more sensitively than that of LDH and cell viability. Meanwhile ,the activity of Na +,K + ATP enzyme decreased gradually. The rate of apoptosis of myocardial cells significantly changed for 180 min after treated with SOV at 100 ?mol?L -1 and 1 mmol?L -1 . CONCLUSION Guinea pig ventricular myocardial cells were only injured when exposed for 30 min to 100 ?mol?L -1 and 1 mmol?L -1 concentrations of SOV higher than the effective concentration of the drug.
6.Establishment of an blood-brain/tumor barrier modal in vitro by coculturing brain endothelial cells,pericytes and U251
Haiyang YANG ; Li CAI ; Jiang LONG ; Jihong HE
Chongqing Medicine 2014;(20):2613-2616,2619
Objective To establish a stable in vitro model of blood-brain/tumor barrier (BBB/BTB) simulating in vivo state u-sing the human brain endothelial cells(hBMEC) ,brain pericytes (PC) and U251 glioma cells(U251) .Methods An in vitro model of BBB/BTB was constructed using Transwell inserts(pore size 1 .0 μm)coculture while three cell subculture reaches a certain num-ber .Its barrier function was evaluated by the 4-hour leakage test ,inverted microscope to observe morphological changes ,horseradish peroxidase permeability test and tight junction protein Claudin-5 and Occludin identification expression observed by immunofluores-cence technology .Results hBM EC monolayers demonstrated a typical cobblestone-like appearance ,Non-contact spindle cocultured monolayer growth appeared its uniqueswirl poolshape ,fusiform morphology ;the pericytes displayed irregular shape and overlap-ping grow ;U251 showed typical tumor cell growth .Fluid leakage test were interviewing a certain level difference in three different ways to build models ;immnocytochemical staining showed a continuous and dense tight junction formed between the endothelial cells but intensity ranging ;horseradish peroxidase permeability of three different models showed that pure hBMEC (43 .490 ± 3 .572)% ,hBMEC+U251(36 .540 ± 1 .475)% ,hBMEC+ PC+U251(26 .460 ± 2 .372)% was significantly lower ,the difference was statistically significant(t=19 .330 ,P<0 .01) .Conclusion hBMEC+ PC+ U251 cocultured the morphology ,tructure and barrier function have more advantages than pure hBMEC and hBMEC+U251 ,have the basic characteristics and function of BTB/BBB ,and they have provided a new tool reseach function ,regulation mechanisms and drug screening the BTB/BBB in future .
7.Comparison of multi-slice spiral CT portography and endoscopy in the diagnosis of esophageal and gastric varices in liver cirrhosis
Xin HE ; Zhongkui HUANG ; Liling LONG ; Qijun WEI ; Xiaohong JIANG ; Pingping GUO ; Chao XIANG ; Jiang LAN
Chinese Journal of Radiology 2012;(12):1092-1095
Objective To study the correlation of multi-slice CT portography (MSCTP) and digestive endoscopy in the diagnosis and evaluation of esophageal and gastric varices (EGV) caused by cirrhosis.Methods A total of 92 patients with cirrhosis were enrolled in the prospective study.All the patients were examined by endoscopy and 64-slice spiral CT scan in 4 weeks.The types,grading of EGV were observed by endoscopy and MSCTP,and Kappa conformance test was applied with the endoscopic findings as gold standard.The sensitivity,specificity,consistency,and Youden index were evaluated for the diagnosis of sophageal and gastric varices by MSCTP.Results Sixty-five patients were diagnosed to have EGV by endoscopy and 27 were negative.The positive patients included 45 patients of GOV1,19 of GOV2 and 1 patient of IGV1.MSCTP diagnosed 67 cases of EGV and 25 patients of negative results.The positive patients included 46 of GOV1,18 of GOV2 and 3 of IGV1.Two patients of IGV1 varicose veins without positive findings on endoscopy were diagnosed by using MDCTP,which revealed isolated varicose veins under the gastric mucosa.There was high consistency between MSCTP and EGV in the diagnosis of EGV (Kappa =0.732,P < 0.01).The sensitivity of MSCTP was 93.8%,specificity was 77.8%,consistency was 89.1%,and Youden index 71.6%.There was high consistency between MSCTP and EGV in the classification of EGV (Kappa values were 0.743 and 0.763,P < 0.01).Conclusions There is high consistency between MSCTP and digestive endoscopic in the diagnosis and classification of EGV in cirrhosis.MSCTP is superior to endoscopy in the detection of gastric varices.
8.Changes in learning and memory ability and brain cholinesterase activity in the rats with coal burning fluorosis
Chuan-zhi, GUI ; Long-yan, RAN ; Chang-xue, WU ; Yi-guo, LONG ; Jiang, HE ; Hua, ZHANG ; Zhi-zhong, GUAN
Chinese Journal of Endemiology 2009;28(5):497-500
Objective To observe the influence of coal burning fluorosis on learning and memory ability in rats and reveal its possible mechanisms. Methods Healthy 48 SD rats were divided into control, low-fluoride and high-fluoride group. All rats in fluoride exposed groups were fed with the eom polluted by drying processes with burning coal containing high level of fluoride obtained from the endemic fluorosis area to produce the animal model of fluorosis. The experiment period were 3,6 mouths, respectively. The ability of leaning and memory was measured by Morris test and cholinesterase activity detected by photometric method at 3 or 6 month after experiment, respectively. Results Fluoride contents signifieantlly influenced the escape latency, the numbers of crossing the platforms and the time of staying the platforms(the value of F was 29.29,6.47,6.50, respectively, P<0.01).In addition, the numbers of crossing the platforms and the time of staying the platforms were influenced by the exposed time(the value of F was 16.11,45.59, P<0.01). Furthermore, the fluoride contents and the exposed time had an interaction between the numbers of crossing the platforms and the time of staying the platforms (the value of F was 4.67,5.68, P<0.05 or<0.01). Three months after the experiment, the mean values of escape latency [(14.71± 4.85)s] of rats in highly fluoride exposed group were significantly prolonged as compared with controls [(9.28±4.22)s]; 6 month after the experiment, the mean values of escape latency[(12.42±8.03)s, (17.48± 8.05)s] of rats in both groups exposed to fluoride were significantly prolonged as compared to controls [(7.04± 3.29)s, P<0.05]. The decreased numbers of crossing the platforms[(1.62±0.87)number] and the declined time of staying the platforms[(16.70±5.02)s] were found in the rats exposed to high fluoride as compared to controls [(3.53±1.67 )number, (23.33±5.35)s, P<0.05]. The fluoride contents obviously influenced the activities of acetylcholinesterase and butylcolinesterase (the value of F was 12.83,13.27, P<0.01). On the other hand, the times of breeding also influnced the activities of butylcolinesterase (the value of F was 16.26, P<0.01). In 3 months of the experiment, the activities of butylcolinesterase [(0.55±0.12)kU/g] in low fluoride exposed group were significantly decreased in comparison with controls[(0.73±0.10)kU/g, P<0.05]. The activities of acetylcholinesterase[(0.62±0.42)kU/g] and butylcolioesterase[(0.58±0.10)kU/g] in high fluoride group were significantly decreased as compared to eontrois[(1.41±0.52), (0.73±0.10)kU/g, P<0.05]. The correlation analysis showed that there was a negative correlation between the cholinesterase and the escape latency(r=-0.68, P< 0.01), and a positive correlation between the cholinesterase and the time of staying the platforms(r=0.57, P< 0.01). Conclusions The ability of learning and memory in rats with coal buring fluorosis was decreased, which might be connected to the decreased activity of cholinesterase in a dose-effect correlation.
9.Distortion Product Otoacoustic Emissions in Normal Infants Hearing Screening
bing, LIAN ; yan-chun, SHAN ; ji-qun, WANG ; li-xin, JIANG ; long, HE
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To investigate the usage of distortion product otoacoustic emissions(DPOAE) in normal infants hearing screening.Methods One hundred infants(200 ears) without clinical condition of illness were checked in GSI 70 DPOAE screening.The pure tones were 2,3,4 kHz,the OAE screener scored the test result as a pass,when the responses were above the PASS/REFER line and the signal-to-noise was at least 10 dB.Results Ninty-four infants(188 ears) passed hearing screening. Four infants(8 ears) with ceruminous impaction didn′t pass screening;after extracting impacted cerumen from the external auditory meatus,the 4 infants(8 ears) passed screening.Two infants(2 ears) with secretory otitis media didn′t pass screening,but after medicine treatment for 1 week,they passed screening too.Conclusion DPOAE has advantages in infants hearing screening.J Appl Clin Pediatr,2006,21(3):168-169
10.The effect of Ulinastatin on the delivery of cytokines in patients with septic shock
Long-Yuan JIANG ; Lian-Hong YANG ; Jian-Xing CHANG ; Zhi-Jiang HE ; Tao YU ; Zi-Tong HUANG ;
Chinese Journal of Emergency Medicine 2006;0(12):-
Objective To investigate the effect of Ulinastatin on the delivery of cytokines in patients with septic shock.Methods It was a prospective and controlled clinical study.Seventy-eight patients with septic shock were randomly divided into control group and treatment group and thirty-nine in every group.Patients in treatment group received Ulinastatin 200 000 units intravenous everyday for 3 days,while those in control group received equal volume of normal saline as placebo.At different time points (at 24 th,48 th,72 th hour after start of treatment),the levels of tumor necrosis factor-alpha (TNF-?),interleukin-1 (IL-1),interleukin-6 (IL-6 ),interleukin-8 (IL-8) and superoxide dismutase (SOD) in serum were assayed.Results In comparison with control group,the levels of TNF-?,IL-1,IL-6,IL- 8 of treatment group decreased markedly (P<0.05,P<0.01) at different time points,whereas the level of SOD was higher markedly (P<0.05,P<0.01) at various time points.Conclusion Ulinastatin has protective effect on patients with septic shock through decreasing the levels of TNF-?,IL-1,IL-6,IL-8 and increasing in the level of SOD.