2.Role of AMPK-dependent autophagic signaling pathway in ketamine-induced reduction of diabetic neuropathic pain in rats
Jie DING ; Yimin HU ; Chun YANG ; Bin ZHU ; Fei HUA
Chinese Journal of Anesthesiology 2016;36(2):180-182
Objective To evaluate the role of AMP-activated protein kinase (AMPK)-dependent autophagic signaling pathway in ketamine-induced reduction of diabetic neuropathic pain (DNP) in rats.Methods Sixty male Wistar rats,aged 3 months,weighing 200-250 g,were equally randomized into 5 groups using a random number table:control group (C group),normal saline group (NS group),ketamine group (K group),ketamine + Compound C group (KC group),and ketamine + 3-methyladenine (3-MA) group (KM group).DNP model was established by intraperitoneal injection of streptozocin (STZ)65 mg/kg in anesthetized rats.Four weeks later,the equal volume of normal saline,ketamine 10 mg/kg,ketamine 10 mg/kg + Compound C1 mg/kg,and ketamine + 3-MA 2 μl were injected intraperitoneally once a day for 7 consecutive days in NS,K,KC and KM groups,respectively.The mechanical paw withdrawal threshold (MWT) was measured on 8th day.The rats were then sacrificed,and the lunbar segment (L1-5) of the spinal cord was removed for determination of the expression of AMPKαt,Beclin-1,microtubule-associated protein 1 light chain (LC) 3B (by Western blot) and dendritic spine density in the dorsal root ganglia.Results Compared with group C,the MWT,expression of AMPKα,Beclin-1,and LC3B,and dendritic spine density were significantly decreased in group NS (P<0.05).Compared with group NS,the MWT,expression of AMPKαt,Beclin-1,and LC3B,and dendritic spine density were significantly increased in group K (P<0.05).Compared with group K,the MWT,expression of AMPKα,Beclin-1,and LC3B,and dendritic spine density were significantly decreased in KC and KM groups (P<0.05).Conclusion Activation of AMPK-dependent autophagic pathway is involved in ketamine-induced reduction of DNP in rats.
3.Developmental Characters of Neural Stem Cells in Occipital of Cortex from Human Fetal Brain at Different Ages
bo, HU ; ai-hua, LI ; yu-lin, AN ; zhi-chun, FENG
Journal of Applied Clinical Pediatrics 2006;0(14):-
Objective To investigate the developmental characters of neural stem cells(NSCs) in occipital of cortex from human fetal brain at different age.Methods Ninety cases of embryoes at gestational age 16-32 weeks and by induction of labor with water bag were collected for determining distribution,shapes,growth modes and the number of NSCs in the occipital of cortex with immunohisto- chemical method under light microscope.Results It was noted that NSCs existed in the occipital of cortex from human fetal brain at different ages.NSCs mainly distributed in layers of cone cells and inner granule cells.NSCs existed in the occipital of cortex of different fetal age included middling round cells,NSCs had enations from 0 to 1.Nucli were larger than plasm.Each NSC had nucleoli from 2-4 and rarefaction chromatin.Most of NSCs distributed in three growth modes including crowd,cluster and clone,occasionally with a single growth mode among other nerve cells.There were no differences including distribution,shapes,growth modes and the number of NSCs in the occipital of cortex between groups,but,NSCs gradually decreased with increasing of age.Conclusion NSCs exists in the occipital of cortex from different gestational age,and the number of NSCs decreases with increasing of age.
4.Effects of Munc13-1 and Munc18-1 on dopamine secretion dysfunction in manganese-exposed SH-SY5Y cells
Changzhe LI ; Chun YU ; Hua ZHAO ; Jun LI ; Ting HU
Journal of Environmental and Occupational Medicine 2023;40(3):268-272
Background Neurotransmitter secretion disorder induced by chronic manganese poisoning has always been one of the important causes of body injury, but the mechanism of neurotransmitter secretion disorder caused by manganese is not clear at present. Objective To investigate the effects of presynaptic membrane intracellular protein 13-1 (Munc13-1) and synapse fusion protein binding protein 18-1 (Munc18-1) on dopamine secretion dysfunction induced by manganese chloride (MnCl2) in human neuroblastoma (SH-SY5Y) cells. Methods A SH-SY5Y cell model induced by MnCl2 was established. Cell viability was measured by MTT assay. Four experimental groups were set up: control group and low-, medium-, and high-dose manganese groups (0, 100, 200, and 400 μmol·L−1 MnCl2). They were treated with corresponding doses of MnCl2 for 24 h. The secretion of dopamine was measured by enzyme-linked immunosorbent assay. The mRNA expression of Syntaxin-1 was detected by real-time quantitaive PCR. Total cell proteins were extracted, and the protein expression levels of Munc13-1, Munc18-1, and Syntaxin-1 were detected by Western blotting. The correlations of MnCl2 exposure and dopamine secretion with the protein expressions of Munc13-1 and Munc18-1 were also analyzed by Pearson correlation. Results Compared with the control group, the cell viability rate decreased gradually with the increase of manganese exposure concentration, and the difference between the medium- and the high-dose manganese groups was statistically significant (P<0.05). The concentration of dopamine in cell culture medium of all manganese exposure groups decreased with the increase of manganese concentration, and compared with the control group and the low-dose manganese group, the medium- and the high-dose manganese groups were statistically significant (P<0.05). The expression of Syntaxin-1 at mRNA or protein level did not change significantly among groups (P>0.05). Compared with the control group, the protein expression of Munc13-1 decreased and that of Munc18-1 increased with the increase of manganese concentration (P<0.05). Compared with the low-dose manganese group, the changes of Munc13-1 protein in the high-dose manganese group and Munc18-1 protein in the medium- and high-dose manganese groups had statistical significance (P<0.05). Compared with the medium-dose manganese group, the protein changes of Munc18-1 in the high-dose manganese group were statistically significant (P<0.05). The correlation analysis showed that MnCl2 dose was negatively correlated with Munc13-1 protein expression (r=−0.898, P<0.05), and positively correlated with Munc18-1 protein expression (r=0.678, P<0.05). Dopamine secretion was positively correlated with Munc13-1 protein expression (r=0.932, P<0.05), and negatively correlated with Munc18-1 protein expression (r=−0.817, P<0.05). Conclusion The inhibition of dopamine secretion in SH-SY5Y cells induced by manganese exposure is related to up-regulation of Munc18-1 and down-regulation of Munc13-1 expression levels, which may be one of the reasons for nerve injury caused by manganese.
5.Correlation between invasion and metastasis of gastric cancer and serum angiopoietin-2
Shao-ping LIU ; Chun-hua FANG ; Li-qun YU ; Xian-feng ZHANG ; Ya-hua HU
Chinese Journal of Postgraduates of Medicine 2011;34(28):31-34
Objective To study the clinical significance of the serum angiopoietin-2(Ang-2) in the diagnosis,recurrence,invasion and metastasis of gastric cancer.Methods The serum Ang-2 and carcino-embryonic antigen (CEA) levels in 158 patients with gastric cancer (gastric cancer group) and 30 normal controls(control group) were measured by enzyme linked immunosorbent assay(ELISA) technique respectively.The serum Ang-2 and CEA levels were also measured 2 weeks after operation in gastric cancer group and reexamined in the recurred gastric cancer patients in 2 years after operation (recurred and metastasis group).The correlation between the serum Ang-2 level and pathologic c haracterization of gastric cancer was evaluated.Results The serum Ang-2 and CEA levels in gastric cancer group [ (331.8 ±64.3),(42.6 ±37.3)μg/L] and recurred and metastasis group [(318.7 ±72.9),(40.5 ±36.7)μg/L] were significantly higher than those in control group [ (187.4 ± 32.7),(4.2 ± 3.1 )μ g/L] (P < 0.01 ),and the serum Ang-2 level 2 weeks after operation [ (211.6 ± 75.1 ) μ g/L ] was significantly decreased to the control group (P > 0.05 ),while the serum CEA level [ (33.4 ± 30.6) μ g/L ] was still significantly higher than the control group (P < 0.01 ).The sensitivity of the serum Ang-2 for diagnosis of gastric cancer was markedly higher than that of the serum CEA (P < 0.01 ).There was correlation between serum Ang-2 and degree of tumor differentiation,TNM pathological staging,lymphatic metastasis,invasion depth and tumor size (p <0.01 ),but there was no correlation between serum Ang-2 and tissue classification and location of gastric cancer (P> 0.05).Conclusion The serum Ang-2 level is suggested to be a valuable gastric cancer marker and conduce to the diagnosis of gastric cancer,the monitoring of recurrence after operation and evaluation of prognosis.
6.The effect of exposure to 50 Hz magnetic fields on the proliferation of different cell densities of human MG-63 osteosarcoma cell line in vitro
Wen-Chun ZHAO ; Hua WU ; Wei-Ming MA ; Hai-Hu HAO ; Hai-Jun ZHANG ; An HU ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(05):-
Objective To study the effects of 50 Hz magnetic fields on the in vitro proliferation of human osteosareoma cell line MG-63 with different cell densities.Methods Four different magnetic intensities(1 mT, 2 mT,3 roT,4 mT)were used to stimulate the cells,and the experiment was repeated with different cell densities. The method of MTT was employed to evaluate the level of proliferation.Results Fifty Hz magnetic fields signifi- cantly affected the level of proliferation of human osteosareoma cell line MG-63,and the 2 mT intensity exerted the greatest influence on it.The effects of the magnetic field differed with different cell densities.Conclusion The effect of 50 Hz magnetic fields on the in vitro proliferation of human osteosarcoma cell line MG-63 was not only relat- ed to the magnetic intensity,but also the cell density,
7.The effect of brain-derived neurotrophic factor on the angiogenesis.
Chun-yan SUN ; Yu HU ; Tao WU ; Ya-dan WANG ; Hua-fang WANG ; Wen-juan HE
Chinese Journal of Pathology 2006;35(4):238-239
Animals
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Brain-Derived Neurotrophic Factor
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pharmacology
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Cell Movement
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drug effects
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Cell Proliferation
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drug effects
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Cells, Cultured
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Chick Embryo
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Chorioallantoic Membrane
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blood supply
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Endothelial Cells
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cytology
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drug effects
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physiology
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Female
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Humans
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Mice
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Mice, Inbred C57BL
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Neovascularization, Physiologic
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drug effects
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Vascular Endothelial Growth Factor A
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pharmacology
8.Study on HPLC fingerprint of jinzhen oral solution.
Jun-Hua HU ; Qian-Qian SU ; Ting-Ting LI ; Jia-Chun LI ; Wei XIAO
China Journal of Chinese Materia Medica 2014;39(19):3768-3771
The HPLC fingerprint determination method of Jinzhen oral solution was established to provide a new method for quality control of Jinzhen oral solution. RP-HPLC was used for phenomenex Luna C18 (4.6 mm x 250 mm, 5 μm) chromatographic column, with 0.1% H3 PO4 water solution and acetonitrile as the mobile phase for gradient elution. The detection wavelength was 280 nm. HPLC fingerprint of Jinzhen oral solution was established to identify 17 common peaks in Jinzhen oral solution. The similarity of fingerprints of 10 batches of finished products was more than 0. 90. The established HPLC fingerprint has a better precision, reproducibility and stability, and can be applied in quality control of Jinzhen oral solution.
Chromatography, High Pressure Liquid
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methods
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Drugs, Chinese Herbal
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chemistry
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Quality Control
9.Construction of mouse models of experimental allergic rhinitis and the correlation with Th1/Th2 imbalance
Linge LI ; Juan FENG ; Bin HU ; Xi SHOU ; Chun ZHANG ; Yu ZHANG ; Hua ZHANG
Chinese Journal of Tissue Engineering Research 2015;(40):6515-6519
BACKGROUND:Atopic individuals predispose to alergic rhinitis after contacting with an alergen, which is mainly released by IgE-mediated histamine. Alergic rhinitis is a kind of non-infectious inflammatory disease that involves a variety of immune cels and cytokines and is related to Th1 and Th2 immune imbalance. Interferon γ is a cytokine secreted by Th1 cels, while interleukin-4 is a cytokine secreted by Th2 cels. OBJECTIVE:To establish a stable 129Sv mouse model of alergic rhinitis, so as to lay the foundation of establishing gene knockout 129Sv mouse models of alergic rhinitis, and to observe the concentration variation of IgE, interleukin-4 and interferon γ. METHODS: Twenty-four mice were randomly divided into 2 groups: model and control groups. Mice in the model group were sensitized by intraperitoneal injection of ovalbumin to establish mouse models of alergic rhinitis. Mice in the control group were subjected to intraperitoneal injection of PBS. After successful modeling, pathological changes of nasal eosinophils and plasmocyte infiltration were evaluated by nasal mucosa staining. Interleukin 4 and interferon γ levels and ovalbumin-specific IgE antibody concentration were detected by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:Enzyme-linked immunosorbent assay showed that compared with the control group, serum ovalbumin-IgE and interleukin-4 concentrations were significantly increased, and Interferon γ concentration was significantly lower in the model group (P < 0.05). Hematoxylin-eosin staining results showed that cilia lodging and submucosal serous gland hyperplasia, eosinophils and plasmocyte infiltration were obvious. These results confirm that a mouse model of allergic rhinitis was successfully constructed in this study and the pathogenesis of allergic rhinitis was related to Th1/Th2 imbalance.
10.Phenotype of congenital cataract accompanied with iris dysplasia
Chun-ling, WEI ; Juan-juan, LI ; Zhu-lin, HU ; Hua, LI
Chinese Journal of Experimental Ophthalmology 2012;30(7):630-632
Background Certain relationship has been found between phenotype and genes mutation of congenital cataract.It is clear that different genetic mutations can cause the same complication in congenital cataract,meanwhile,different complications may be caused by the same gene mutation.However,their mechanism is still remained unclear.Objective This study was to observe the phenotype of congenital cataract accompanied with iris dysplasia.Methods Fifteen patients with congenital cataract accompanied with iris dysplasia were included in this study.The slit lamp,gonioscope and ophthalmoscope were used for the examination of the anterior ocular segment,the anterior chamber angle and fundus on all the patients.This study was approved by the Ethic Committee of Second People' s Hospital of Yunnan Province.Written informed consent was obtained from each patient or the custodian prior to any medical procedure.Results All the patients showed binocular involvement.Congenital nuclear cataract with whole coloboma of iris was seen in 7 cases,and 2 cases showed an entire cataract associated with incomplete coloboma of iris.Entire cataract with aniridia was diagnosed in 5 patients,and suture cataract complicated with aniridia was in 1 patient.Conclusions Some regular patterns can be implied between the morphological type of cataract and iris dysplasia,which may be helpful for further study of these diseases.