1.Mitochondrial DNA point mutations studies in hereditary ataxia
Chinese Journal of Practical Internal Medicine 2001;0(10):-
Objective To study the possible relationship between mitochondrial DNA(mtDNA)and hereditary ataxia(HA).Methods Polymerase chain reaction(PCR)was used to amplify 4 mtDNA segments of 30 patients with HA、some of their relatives and 35 volunteers.The point 3243、8993、8344 and point 11778 lied in the 4 mtDNA segments respectively.For the PCR products of point 3243 and 8993,restriction fragment length polymophism(RFLP)was performed to search for A3243G、T8993G or T8993C point mutations.For point 8344 and 11778 PCR products,single strand conformation polymorphism(SSCP)was executed to detect mutations.The HA patients’ results of SSCP were compared with their relatives and volunteers’.Sequencing would be carried out to find out exact mutations in those subjects whose SSCP results were abnormal.Results We had not found the A3243G、T8993G or T8993C point mutations in our study.All subjects’ mtDNA segments of point 8344 had not been found mutations.However,a new mtDNA point mutation-A11893G-was identified in 2 patients and 1 relative without symptoms from pedigree 1.Conclusion This new point mutation of mtDNA might be related to HA.
2.Interhemispheric functional alterations in unilateral temporal lobe epilepsy
Chinese Journal of Nervous and Mental Diseases 2016;42(10):616-622
Objectives To investigate the alterations of homotopic connectivity in unilateral temporal lobe epilepsy (TLE). Methods Resting-state functional MR imaging data were acquired from patients with unilateral temporal lobe epi?lepsy (n=43) [21 left temporal lobe epilepsy(LTLE) and 22 right temporal lobe epilepsy(RTLE)] and normal controls (NC) (n=20). The functional connectivity between any pair of symmetric interhemispheric voxels (i.e., functional homotopy) was measured by voxel-mirrored homotopic connectivity (VMHC). Results Compared with NC group, the LTLE patients showed decreased functional connectivity in the bilateral supplementary motor areas(t=-3.35), middle temporal gyri (t=-3.50), medial frontal gyri(t=-3.20)and inferior parietal lobules(t=-3.66), and increased interhemispheric func?tional connectivity in the bilateral angular gyri(t= 3.03), occipital gyri(t= 4.25) and superior parietal gyri(t= 3.62). Compared with control subjects, RTLE patients exhibited decreased functional connectivity in the bilateral middle tempo?ral gyri(t=-4.26), precentral gyri(t=-3.29), as well as increased interhemispheric functional connectivity between bi?lateral inferior occipital gyri(t=3.22),parahippocampa gyri(t=3.32), cerebellum(t=3.50). Conclusions There are al?terations of the functional synchronization and coordination in the two hemispheres in unilateral TLE patients.
3.Interhemispheric Functional and Structural Alterations in Unilateral Temporal Lobe Epilepsy
Chinese Journal of Medical Imaging 2016;24(12):884-889
Purpose To investigate the alterations of homotopic functional connectivity and interhemispheric structural connectivity in patients with unilateral temporal lobe epilepsy (unilateral TLE).Materials and Methods Resting-state functional MR imaging data were acquired from patients in the first affliated hospital of Guang Zhou Medical University who were enrolled from Oct.2013 to Oct.2015 with unilateral temporal lobe epilepsy (n=43) [21 left TLE(LTLE) and 22 right TLE(RTLE)] and normal controls (NC) (n=24).We mainly concerned about the functional connectivity between any pair of symmetric interhemispheric voxels which were measured by voxel-mirrored homotopic connectivity (VMHC).Structural magnetic resonance images were acquired in 10 LTLE patients,16 RTLE patients and 20 NCs.Homotopic regions showing abnormal functional connectivity in patients were adopted as regions of interest for DTI.The FA values were compared between groups.Two-sample t test were used.Results Compared with NC group,the LTLE patients showed increased interhemispheric functional connectivity among the bilateral angular gyri,occipital gyri and superior parietal gyri,and decreased functional connectivity between the bilateral supplementary motor areas,middle temporal gyri,medial frontal gyri and inferior parietal lobules.Compared with control subjects,RTLE patients exhibited increased interhemispheric functional connectivity among bilateral inferior occipital gyri,parahippocampa gyri,cerebellum,as well as decreased functional connectivity among the bilateral middle temporal gyri,precentral gyri and inferi or frontalgyri.Statistical results indicated that the FA value of the commissural fiber bundles connecting the bilateral hippocampal gyri was smaller in RTLE patients than NC patients (t=10.55,P<0.01).The FA value of the association fibers connecting the bilateral hippocampal gyrus had no significant difference between the LTLE group and NC group.Conclusion The increase or decrease of VMHC value in some brain regions in TLE suggested that there were changes in the functional synchronization and coordination in the two hemispheres in patients with unilateral TLE.In the RTLE patients,some brain regions which had abnormal interhemispheric functional connectivity also had structural damage.
4.The influence of high frequency, repetitive transcranial magnetic stimulation on long-term potentiation in the hippocampus and on spatial learning and memory after global cerebral ischemia
Xiaokuo HE ; Huihua LIU ; Tiebin YAN
Chinese Journal of Physical Medicine and Rehabilitation 2012;(12):883-888
Objective To investigate the effects of repeated high frequency transcranial magnetic stimulation (rTMS) on spatial learning and memory function,and on long-term potentiation (LTP) after global cerebral ischemia and reperfusion,and to explore the mechanisms involved.Methods Eighty-three male Wistar rats were studied.Five were tested to determine their average motor threshold (Tm).The others were divided into a normal control group,a cerebral ischemia and reperfusion model group and an rTMS group.Cerebral ischemia was induced with the four vessel occlusion method for 10 minutes.The rTMS treatment protocol (10 Hz stimulation for 5 s at the resting threshold,twice a day) was applied over a 2-week period from day 3 post-operation.The Morris water maze test was performed to observe spatial learning and memory at post-operation day 2 and day 4.The field excitatory postsynaptic potentials,population spike and the magnitude of long-term potentiation (LTP) induced by theta burst electric stimulation were recorded from the perforant path to the dentate gyrus (PP-DG).Results At post-operation day 3,rats in the untreated cerebral ischemia and reperfusion model group exhibited a significant decrease in the magnitude of the PP-DG LTP as compared to the normal group.No significant difference in LTP was found between the model group and the rTMS group.After the 2 weeks of treatment the LTP levels in the rTMS treated group were significantly higher than in the two untreated groups.In the Morris water maze testing,the average escape latency in the rTMS group was significantly shorter than that of the cerebral ischemia and reperfusion model group (which was not treated).In the probe trials,the time in the original quadrant of the platform and the time of crossing the platform were both significantly less for the rTMS-treated rats than for those not treated.Conclusions High frequency rTMS can improve spatial learning and memory after global cerebral ischemia and reperfusion by enhancing the LTP induced in the hippocampus.High frequency rTMS might exert this beneficial effect by modulating the function of intermediate neurons in the hippocampal neuronal network and by promoting neurotransmitter release.
5.Study of mitochondrial DNA point mutations at positions 3243, 8993 in hereditary ataxia
Jing WANG ; Huihua LIU ; Shuguang LUO
Journal of Clinical Neurology 1988;0(02):-
Objective To study the possible relationship between mitochondrial DNA point mutations and hereditary ataxia (HA). Methods Polymerase chain reaction (PCR), restriction fragment length polymophism (RFLP) were performed to search A3243G, T8993G or T8993C point mutations in the amplified mitochondrial DNA of extract human perpheral white blood cells of 26 patients with HA and 35 normal controls. Results No point mutations of mitochondrial DNA A3243G, T8993G or T8993C were found in HA group and control group.Conclusion mitochondrial DNA A3243G, T8993G and T8993C mutations are not likely to be genetic factors of hereditary ataxia.
6.Protective effect of ulinastatin on the renal ischemia/reperfusion injury in rats
Congcong CHEN ; Ziming LIU ; Huihua WANG
Chinese Journal of Anesthesiology 1997;0(11):-
Objective To investigate the effects of ulinastatin on renal ischemia/reperfusion (I/R) in rats and assess the possible mechanism. Methods Seventy-five male SD rats weighing 210-250 g were randomly divided into three groups with 25 animals in each group : (Ⅰ) control group received sham operation; (Ⅱ)renal I/R group and (Ⅲ) ulinastatin + I/R group. The animals were anesthetized with intraperitoneal ketamine 50 mg?kg-1 . Bilateral kidneys were exposed through midline incision and bilateral renal pedicles were occluded with atraumatic mini-clamp. The kidneys turned pale. In control group the kidneys were exposed but their pedicles were not clamped, while in ulinastatin group (Ⅲ) ulinastatin 12 500 u was given i. v. 30 min before ischemia and at the beginning of reperfusion. Blood samples and kidneys were obtained at 0, 2, 6, 12, 24 h of reperfusion for determination of blood urea nitrogen (BUN) and serum creatinine (Cr) concentrations and the expression of heat shock protein 70 (HSP 70) and bcl-2 in kidney and for microscopic examination. Paller scoring system was used to evaluate the severity of histological changes ( the higher the score, the worse is the change) .Results BUN and serum Cr levels were significantly lower at 12 and 24 h of reperfusion in group Ⅲ than in group Ⅱ (P 0.05) . Conclusion Ulinastatin can reduce the risk of renal dysfunction and injury associated with renal I/ R. The protective effect of ulinastatin may be associated with the up-regulation of bcl-2 expression.
7.The role of angiogenesis in myocardial injury in septic mice
Anlei LIU ; Jie LIU ; Tianpeng ZHANG ; Shubin GUO ; Huihua LI
Chinese Journal of Emergency Medicine 2011;20(12):1295-1299
Objective To investigate the angiogenesis,apoptosis and their mechanisms in septic mice with myocardial injury.Methods Forty male C57BL/6 mice aged 8 weeks were randomly ( random number) divided into two groups:the sepsis group and the control group.The mice of sepsis group were treated with lipopolysaccharide (LPS) ( 10 mg/kg Intraperitoneal injection) while the mice of control group were treated with saline solution instead (10 mg/kg Intraperitoneal injection).Cardiac function of mice (n =40) was evaluated with ultrasound 6 hours after LPS administration.Subsequently,the tissues of heart,lung and kidney of mice (n =6) were taken and treated with Haematoxylin -Eosin staining (H&E) in order to observe the pathological changes and verify the successfulness of modeling.Immunohistochemistry staining with PECAM - 1 and α - SMA was used to identify the angiogenesis in the heart ( n =3 ),while the TUNEL apoptosis assay was applied for detecting the myocardial cell apoptosis ( n =3 ).The mRNA was extracted from heart tissue (n =6) to observe the expression of HIF-1 ot which was proved to be an angiogenesis factor.All the results were analyzed by independent sample t - test.Results Compared to the control group,mice in the sepsis group showed increased in thickness of left ventricular diastolic anterior wall ( t =- 4.60,P < 0.05 ) and thickness of left ventricular systolic anterior wall (t =-3.24,P <0.05 ) along with decrease in left ventricular end diastolic diameter ( t =3.57,P < 0.01 ) and stroke volume ( t =5.51,P < 0.01 ).Immunohistochemistry staining with alpha - SAM antibody revealed increase in cardiac angiogenesis in the sepsis group (t =- 11.00,P < 0.01 ).TUNEL apoptosis assay demonstrated apoptosis of the cardiomyocytes [ sepsis group versus control group:( 191.31 ±5.41 ) vs ( 52.24 ±4.32) ] and RT - PCR showed an increase in the expression of HIF - 1 alpha in the mice of the sepsis group ( t =- 8.12,P <0.05) Conclusions There were apparent myocardial angiogenesis,apoptosis and cardiac dysfunction in septic animal models.HIF-1α might play a role in the angiogenesis pathway.
8.Observation of the effect of Tanreqing Injection on patients with viral pneumonia
Bin DENG ; Huihua LIU ; Qinming LIU ; Wenhuan DENG
Chinese Traditional Patent Medicine 1992;0(01):-
AIM: To observe the effect of Tanreqing Injection on patients with viral pneumonia. METHODS: 80 cases of viral pneumonia were divided into two groups randomly:40 cases of treatment group and 40 cases of contrast group. The two groups were treated with Tanreqing Injection and ribavirin differently, once per day, 10 to 14 days for one treatment period. RESULTS: The total effective rate of treatment group was superior to that of the contrast group. CONCLUSION:Tanreqing Injection is effective on treating viral pneumonia.
9.COMPARATIVE STUDIES ON THE CHARACTERISTICS OF PROLIFERATION AND DIFFERENTIATION OF SPLEEN COLONY FORMING CELLS FROM DIFFERENT SOURCES
Zuze WU ; Minpei LIU ; Huihua XUE ; Renbao ZHU
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
By using single spleen colony transplantation technique and sex chromosome typing as natural cytogenetic markers, we have been able to demonstrate that most spleen colony forming cells in the adult bone marrow or in foetal liver of inbred LACA or C57 mice possess the potential of re-establishment of myeloid and lymphoid lines of cells in lethally irradiated mice. However, most of the spleen colony forming cells in the peripheral [blood of normal mice possess weak or no potentiality to proliferate or re-establish hemopoiesis in lethally irradiated mice. Our results support the notion that the CFU-S population in mice is heterogeneous.From the nature of colony forming cells in the peripheral blood of normal mice, we are convinced that the assessment of CFU-S content relying upon the spleen colonies as the unique sign to indicate the self-renewal ability of a spleen colony forming cell is by no means valid inasmuch as some plu-ripotent . hemopoietic progenitors derived from pluripotent hemopoietic stem cells may have the same ability to form colonies in vivo.
10.The effects of functional electrical stimulation on motor function and on the proliferation, migration and differentiation of neural stem cells in the subventricular zone
Huihua LIU ; Tiebin YAN ; Zhimei TAN ; Xiaokuo HE
Chinese Journal of Physical Medicine and Rehabilitation 2015;37(9):646-652
Objective To investigate the effects of functional electrical stimulation (FES) on motor function and on the expression, proliferation, migration and differentiation of endogenous neural stem cells in the subventricular zone (SVZ) after cerebral ischemia.Methods Middle cerebral artery occlusion (MCAO) was used to induce a model of cerebral ischemia in 108 rats using the modified Zea-Longa method of intraluminal filament occlusion.They were then randomly divided into an FES group, a placebo stimulation group and a control group with 36 cases in each.Superficial FES electrodes were pasted on the paralyzed forelimbs of the rats in the first two groups, though FES treatment was administered only to the FES group beginning on the 3rd day after the MCAO operation.The stimulation was designed to produce extension of the wrist and digits of the paralyzed forelimb.Before, and after 1,3, 7 and 14 days of the treatment, the neurological deficit was evaluated using modified neurological severity scoring (mNSS).BrdU +/GFAP+, BrdU+/DCX+ and BrdU+/NeuN + cells in the SVZ were detected using immunofluorescence technique.Results After 7 and 14 days of treatment, the average motor function of the rats in the FES group had improved significantly when compared with the averages of the other two groups.Compared with the other two groups, the average number of BrdU +/GFAP+ positive cells in the ischemic SVZ was also significantly greater in the FES group after 7 and 14 days of treatment.After 14 days, BrdU +/Dcx + positive cells in the FES group had also increased significantly more,but only a few BrdU +/NeuN + cells had appeared in any of the three groups.Conclusion FES can improve motor function after cerebral ischemia, and promote proliferation and differentiation of neural stem cells in the SVZ.