2.Quantitative analysis of synaptic vesicle release and readily releasable pool size in hippocampal neurons.
Shu-Jun XU ; Ping WANG ; Di XIA
Acta Physiologica Sinica 2009;61(6):505-510
In central nervous system only a limited number of vesicles exist in the presynaptic terminals. The size and fusion modes of the vesicles were particularly important because of their potential impact on neuronal communications. Efficient methods were needed to analyze the recycling kinetics of synaptic vesicle and the size of readily releasable pool (RRP). In this study, fluorescent dyes with different affinity for membranes (FM1-43 with high affinity and FM2-10 with low affinity) were used to stain the functional synaptic vesicles of cultured hippocampal neurons and the kinetics of vesicle recycling was measured. The results showed that the destaining proportion was larger for FM2-10 than that for FM1-43 during the first trial, while it was greater for FM1-43 than FM2-10 during the second and third trials (first round, 93.0%+/-5.9% versus 57.9%+/-3.5% for FM2-10 and FM1-43, respectively, P<0.0001; second round, 1.4%+/-3.8% versus 24.0%+/-2.3%, P<0.0001; third round, 2.3%+/-1.6% versus 8.6%+/-1.5%, P=0.005). The results indicated that rapid endocytosis existed not only in the first round but also occurred when the vesicles were reused. Moreover, Both high-frequency stimuli and hypertonic sucrose stimuli were used to estimate the RRP sizes in the mix cultured hippocampal inhibitory neurons at 13-14 days in vitro (DIV). We found that the RRP size estimated by hypertonic sucrose stimuli [(200+/-23.0) pC] was much larger than that estimated by high-frequency stimuli [(51.1+/-10.5) pC]. One possible reason for the discrepancies in RRP estimates is that in mix cultured conditions, one neuron may receive inputs from several neurons and hypertonic sucrose stimuli will cause RRP of all those neurons release, while using dual patch recording, only the connection between two neurons was analyzed. Thus, to exclude out the impacts of inputs numbers on RRP sizes, it is more reasonable to use high-frequency stimuli to estimate the RRP size in mix cultured neurons.
Cells, Cultured
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Endocytosis
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Hippocampus
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cytology
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Neurons
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physiology
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Synaptic Vesicles
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physiology
3.Genome amplification and sequence analysis of human bocavirus 2.
Huan-Di CUI ; Yu JIN ; Guang-Cheng XIE ; Wei-Xia CHENG ; Zhao-Jun DUAN
Chinese Journal of Virology 2014;30(3):257-262
To obtain the genome sequence of human bocavirus 2 (HBoV2), different regions of HBoV2 genome were amplified through PCR in fecal specimens which had been identified as single-positive for HBoV2 in 2010. A genome sequence of HBoV2 (HBoV2-NC, 5444 bp) was obtained after sequence assembly. The phylogenetic analysis showed that HBoV2-NC had the closest evolutionary relationship with HBoV2 Lanzhou strain. The predication of inverted terminal repeats of HBoV2-NC by DINAMelt showed that inverted terminal repeats were contained in HBoV2-NC 5' terminal, which had the typical stem-loop structure in other parvoviruses. Finally, some flanking sequences of HBoV2-NC were amplified by linker-PCR.
Base Sequence
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Gene Amplification
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Genome, Viral
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Human bocavirus
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chemistry
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classification
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genetics
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Humans
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Molecular Sequence Data
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Nucleic Acid Conformation
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Parvoviridae Infections
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virology
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Phylogeny
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RNA, Viral
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chemistry
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genetics
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Terminal Repeat Sequences
4.Diagnosis Performance of 99mTc-MIBI and Multimodality Imaging for Hyperparathyroidism
ZHOU JUN ; LU DI-YU ; XIA LIANG ; CHENG XIAO-JIE
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(4):582-586
This study aimed to examine the diagnosis performance of 99mTc-methoxyisobutylisonitrisonitrile (99mTc-MIBI) and multimodality imaging [ultrasound,single-photon emission computed tomography/computed tomography (SPECT/CT)] for hyperparathyroidism (HPT).From Nov.2009 to Dec.2015,clinical data of a total of 43 HPT patients (16 males and 27 females;26-70 years old,average age:51.60±10.66 years old) were retrospectively analyzed.Among them,19 patients with primary hyperparathyroidism (PHPT) underwent 99mTc-MIBI planar imaging,24 [15 with PHPT and 9 with secondary hyperparathyroidism (SHPT)] underwent SPECT/CT hybrid imaging,and 41 (33 with PHPT and 8 with SHPT) had neck ultrasound imaging.Final diagnosis was determined by pathological examination after surgery.The positive rate was compared between different imaging modalities,and the correlation analysis was conducted between imaging results and lesion size or serum parathyroid hormone (PTH) level.The results showed that the total positive rates of 99mTc-MIBI imaging,ultrasound,and the two combined imaging in the 43 HPT cases were 90.70% (39/43),58.54% (24/41),and 100% (41/41),respectively.According to lesion numbers,the positive rates were 79.10% (53/67),53.23% (33/62),and 88.71% (55/62),respectively.SPECT/CT hybrid images were positive in all the 24 patients who underwent this examination.The mean maximum diameters of the lesions in 99mTc-MIBI positive and negative patients were 1.96±0.95 cm and 1.36±0.67 cm respectively,with statistically significant difference noted (P=0.03).The T/NT of 99mTc-MIBI imaging at the early phase was correlated positively with serum PTH level (r=0.40,P=0.01).The T/NT of 99mTc-MIBI imaging at both the early phase and the delay phase was correlated positively with lesion size (r=0.51,and r=0.45,respectively;P<0.01 for both).It was concluded that 99mTc-MIBI imaging presents significant value for location diagnosis of HPT,especially when combined with SPECT/CT hybrid imaging or ultrasound.The 99mTc-MIBI uptake correlates positively with serum PTH level and lesion size.
5.Observation on effect of compound danshen droplet-pill combined with trimetazidine in treating senile unstable angina pectoris.
Zhong-xia QIU ; He-jun MA ; Di-fei WANG
Chinese Journal of Integrated Traditional and Western Medicine 2005;25(9):787-789
OBJECTIVETo investigate the effect of compound Danshen Droplet-pill (DS) combined with trimetazidine (TMZ) in treating senile unstable angina pectoris (SUAP).
METHODSOne hundred and twenty patients with SUAP were eaually and randomly divided into 2 groups, the treated group and the control group. Changes of angina, occurrence of arrhythmia, myocardial infarction and sudden death, myocardial ischemia in ECG and partial indexes of heart function were observed.
RESULTSThe total effective rate in the treated group was 78.3%, while that in the control group was 53.3%, comparison of the two groups showed significant difference (P < 0.05). The incidence rate of arrhythmia in the two groups was 18.2% and 30.0% respectively and that of acute myocardial infarction and sudden death was 0 and 5.0% respectively, also showed significant difference between them (P < 0.05). Condition of myocardial ischemia revealed in ECG and partial indexes of heart function in the treated group were all improved to certain extent. Conclusion DS combined with TMZ is superior in treating SUAP.
Aged ; Aged, 80 and over ; Angina, Unstable ; drug therapy ; Drug Administration Schedule ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Phenanthrolines ; therapeutic use ; Phytotherapy ; Trimetazidine ; therapeutic use ; Vasodilator Agents ; therapeutic use
6.The effects of ultraviolet blood irradiation and oxygenation on oxygen free radical metabolism in rabbits with soman intoxication
Xian-Qing ZHANG ; Xiao-Di ZHANG ; Shi-Jie MU ; Xing LIANG ; Ai-Jun XIA ; Chun-Xu HAI ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(09):-
Objective To investigate the effects of ultraviolet blood irradiation and oxygenation(UBIO)on oxygen free radical metabolism(OFRM)in rabbits with acute soman intoxication.Methods One hundred rabbits were randomly divided into five groups:a control group,a soman intoxication group(I),a soman intoxication plus routine therapy group(TR),a soman intoxication plus UBIO therapy group(UBIO)and a soman intoxication plus complex therapy group(CT).All the rabbits were intervened accordingly.Then the concentrations of malondiade- hyde(MDA)and the activities of superoxide dismutase(SOD),glutathionperoxidase(GSH Px)and catalase (CAT)in serum were determined at 14 d after various treatments.Results Compared with the control group,the concentration of MDA and the activity of CAT in the 1 group were significantly increased(P<0.01),while the activi- ties of SOD and GSH Px were obviously decreased(P<0.05).After UBIO or complex therapy,the serum level of MDA was significantly decreased in comparison with that in the I group(P<0.01),while the concentrations of SOD, GSH Px and CAT were enhanced(P<0.05).Conclusion UBIO therapy can improve antioxidation activity against the injury caused by free radicals and could be used to treat acute soman intoxication,which causes injury from in- creased oxygen free radical concentrations.
7.Umbilical cord blood-derived mesenchymal stem cells ameliorate graft-versus-host disease following allogeneic hematopoietic stem cell transplantation through multiple immunoregulations.
Qiu-Ling WU ; Xiao-Yun LIU ; Di-Min NIE ; Xia-Xia ZHU ; Jun FANG ; Yong YOU ; Zhao-Dong ZHONG ; Ling-Hui XIA ; Mei HONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(4):477-484
Although mesenchymal stem cells (MSCs) are increasingly used to treat graft-versus-host disease (GVHD), their immune regulatory mechanism in the process is elusive. The present study aimed to investigate the curative effect of third-party umbilical cord blood-derived human MSCs (UCB-hMSCs) on GVHD patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and their immune regulatory mechanism. Twenty-four refractory GVHD patients after allo-HSCT were treated with UCB-hMSCs. Immune cells including T lymphocyte subsets, NK cells, Treg cells and dendritic cells (DCs) and cytokines including interleukin-17 (IL-17) and tumor necrosis factor-alpha (TNF-α) were monitored before and after MSCs transfusion. The results showed that the symptoms of GVHD were alleviated significantly without increased relapse of primary disease and transplant-related complications after MSCs transfusion. The number of CD3(+), CD3(+)CD4(+) and CD3(+)CD8(+) cells decreased significantly, and that of NK cells remained unchanged, whereas the number of CD4(+) and CD8(+) Tregs increased and reached a peak at 4 weeks; the number of mature DCs, and the levels of TNF-α and IL-17 decreased and reached a trough at 2 weeks. It was concluded that MSCs ameliorate GVHD and spare GVL effect via immunoregulations.
Adolescent
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Adult
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Cord Blood Stem Cell Transplantation
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methods
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Cytokines
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metabolism
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Dendritic Cells
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metabolism
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Female
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Graft vs Host Disease
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immunology
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therapy
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Hematopoietic Stem Cell Transplantation
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adverse effects
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Humans
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Immunomodulation
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Killer Cells, Natural
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metabolism
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Male
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T-Lymphocyte Subsets
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metabolism
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Transplantation, Homologous
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adverse effects
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Young Adult
8.Betulinic acid prevents alcohol-induced liver damage by improving the antioxidant system in mice.
Jine YI ; Wei XIA ; Jianping WU ; Liyun YUAN ; Jing WU ; Di TU ; Jun FANG ; Zhuliang TAN
Journal of Veterinary Science 2014;15(1):141-148
Betulinic acid (BA), a pentacyclic lupane-type triterpene, has a wide range of bioactivities. The main objective of this work was to evaluate the hepatoprotective activity of BA and the potential mechanism underlying the ability of this compound to prevent liver damage induced by alcohol in vivo. Mice were given oral doses of BA (0.25, 0.5, and 1.0 mg/kg) daily for 14 days, and induced liver injury by feeding 50% alcohol orally at the dosage of 10 ml/kg after 1 h last administration of BA. BA pretreatment significantly reduced the serum levels of alanine transaminase, aspartate transaminase, total cholesterol, and triacylglycerides in a dose-dependent manner in the mice administered alcohol. Hepatic levels of glutathione, superoxide dismutase, glutathione peroxidase, and catalase were remarkably increased, while malondialdehyde contents and microvesicular steatosis in the liver were decreased by BA in a dose-dependent manner after alcohol-induced liver injury. These findings suggest that the mechanism underlying the hepatoprotective effects of BA might be due to increased antioxidant capacity, mainly through improvement of the tissue redox system, maintenance of the antioxidant system, and decreased lipid peroxidation in the liver.
Animals
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Antioxidants/pharmacology
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Blood Chemical Analysis
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Enzymes/blood
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Ethanol/*toxicity
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Lipid Peroxidation/drug effects
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Liver/*drug effects/enzymology/metabolism/pathology
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Male
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Mice
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Random Allocation
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Triterpenes/*pharmacology
10.Comparison of the antagonistic effects of 6 beta-naltrexol and naltrexone against morphine analgesia.
Ling-di YAN ; Ze-hui GONG ; Xia-jun YAO ; Bo-yi QIN
Acta Pharmaceutica Sinica 2003;38(8):578-581
AIMTo compare the antagonistic effects of 6 beta-naltrexol and naltrexone against morphine analgesia.
METHODSThe effects of 6 beta-naltrexol and naltrexone against morphine analgesia were observed in mouse heat radiant tail-flick assay and in mouse (55 +/- 1) degrees C hot plate test. The displacement of 6 beta-naltrexol and naltrexone on binding to CHO-mu receptor was observed by radioligand binding study.
RESULTS6 beta-naltrexol antagonized morphine analgesia but the potency was (6.1 +/- 1.7)% that of naltrexone. The effective duration of 6 beta-naltrexol was 3-4 times that of naltrexone and the peak time of the response was about 0.5-1 h after s.c. equivalent efficacy dose (ED95) in two models. Like naltrexone, 6 beta-naltrexol was effective by oral administration and the potency ratio of p.o./s.c. was 1/3. As an antagonist to opioid receptor, the displacement of 6 beta-naltrexol was about 12.5% that of naltrexone, which was almost in agreement with the efficacies against morphine analgesia in mouse.
CONCLUSIONCompared with naltrexone, 6 beta-naltrexol was less potent but duration was longer.
Analgesia ; Analgesics, Opioid ; antagonists & inhibitors ; Animals ; Female ; Male ; Mice ; Morphine ; antagonists & inhibitors ; Naltrexone ; analogs & derivatives ; pharmacology ; Narcotic Antagonists ; pharmacology ; Pain Threshold ; drug effects ; Receptors, Opioid, mu ; metabolism