1.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.
2.Middle cerebral artery blood flow changes at different tilting angles in patients with cervical and thoracic spinal cord injury
Sheng-jun HU ; Yang HUA ; Gui-ping ZHANG ; Xuemei LI
Chinese Journal of Rehabilitation Theory and Practice 2004;10(9):562-563
ObjectiveTo study the speciality of hemodynamics in middle cerebral artery(MCA) by means of transcranial color Doppler ultrasound(TCCD) at different tilting angles in patients with cervical and thoracic spinal cord injury.Methods48 patients with spinal cord injury were involved. The velocity of peak systolic(PSV), end diastolic(EDV) and mean velocity(MV) of MCA on the tiltable at 0°,30°,60°,80° were measured.ResultsThere was no significant difference in PSV between 0°and 30°(P>0.05), while EDV and MV showed very significant (P<0.01)from 0°to 30°,60°and 80° position. From 0°to 60°,80°,there were differences not only at EDV and MV, but also PSV(P<0.01).ConclusionThe velocities of MCA were changed at different tilting angles in the patients with spinal cord injury.
3. Study on capillary GC fingerprint of Guominxing Biyan Granules
Chinese Traditional and Herbal Drugs 2016;47(6):923-927
Objective: To establish the capillary GC fingerprint of Guominxing Biyan Granules and give a new method for quality control. Methods: The capillary GC was used. The ingredients of common peaks were analyzed by FID and MS. The HP-5 Phenyl Methyl Siloxane (30 m×0.32 mm, 0.25 μm) was used for capillary column. The chromatography conditions were as follows: the temperature of inlet was 200℃ in splitless sampling, the detector temperature was 250℃, the column initial temperature was 50℃, 3℃/min rose to 140℃, keeping 2 min and 10℃/min rose to 200℃, keeping 2 min. Results: Capillary GC fingerprint of volatile oil in Allergic Rhinitis Particles was established. Twelve peaks were selected as the fingerprint peaks of Allergic Rhinitis Particles. The ingredients of common peaks were designated by GC-MS. The similarity between the fingerprint of 10 batches of Allergic Rhinitis Particles Samples were 0.90. Conclusion: The established capillary GC fingerprint has desirable precision, reproducibility, and stability, and can be applied to the quality control of Guominxing Biyan Granules.
4. Determination of cafferic acid and rosmarinic acid in Perilla frutescens leaves and Schizonepeta tenuifolia by HPLC
Chinese Traditional and Herbal Drugs 2015;46(14):2155-2159
Objective: To establish an HPLC method for the simultaneous determination of cafferic acid and rosmarinic acid in Perilla frutescens leaves and Schizonepeta tenuifolia. Methods: The anaiysis was carried out on an Agilent C18 column (250 mm × 4.6 mm, 5 μm), and the mobile phase was composed of actonitrile and 0.1% phosphoric acid aqueous with gradient elution. The detection wavelength was 327 nm. The flow rate 1.0 mL/min at column temperature of 30 ℃. Results: Under the chromatographic condition, cafferic acid and rosmarinic acid were completely separated and other components had no effect on their determination. It showed a good linearity in the range of 0.69-22.12 and 3.65 -116.92 μg/mL. The average recoveries of P. frutescens leaves were 102.9% and 105.6%, and the RSD values were 1.8% and 1.9%. The average recoveries of S. tenuifolia were 101.2% and 101.0% and the RSD values were 2.4% and 1.9%. Conclusion: The established method is rapid, sensitive, accurate, and the test component peak separation degree is good, and could be used for the simultaneous determination of cafferic acid and rosmarinic acid in P. frutescens leaves and S. tenuifolia, which provides a scientific basis for the quality evaluation of P. frutescens leaves and S. tenuifolia.
5.Whole course accelerated hyperfractionation radiotherapy for esophageal carcinoma
Hai-Hua YANG ; Jian-Hua WANG ; Wei-Jun DING ; Wei HU ;
Chinese Journal of Radiation Oncology 2005;0(05):-
Objective To compare the treatment effects and toxicity of convention fractionation ra- diotherapy(CFR),late course accelerated hyperfractionafion radiotherapy(LCAFR) and whole course accel- erated hyperfractionation radiotherapy(WCAFR) for patients with middle or low thoracic esophageal moder- ately differentiated squamous cell carcinoma.Methods Ninety patients with middle or low thoratic esoph- ageal carcinoma were divided into three groups:1.CFR group,patients were irradiated 2.0 Gy/f,5 times a week,to a total dose of 70 Gy;2.LCAFR group,patients were first irradiated with CFR to 40 Gy,then fol- lowed by 1.5 Gy/f bid,at more than 6 hoursinterval,5 times a week,to a total dose of 70 Gy;3.WCAFR group,patients were irradiated 1.5 Gy/f bid,at more than 6 hoursinterval,5 times a week,to a total dose of 72 Gy.Results The 1-,2-,3-year local control rate was 47%,17% and 0% in CFR group,60%,20% and 20% in LCAFR group,60%,40% and 33% in WCAFR group,respectively.The 1-,2-,3-year survival rate was 47%,20% and 10% in CFR group,63%,43% and 17% in LCAFR group,and 63% ,50% and 33% in WCAFR group,respectively.There were better results in the latter two groups(P<0.01).The a- cute toxic effect was severer in the WCAHF and LCAF groups than in CF group.Highter grade 2-3 acute esophagitis rate was observed both in WCAHF and LCAF groups.There were no significant differences in late esophagitis rate between the three groups.The grade 1-2 acute radiation pneumonitis rates were higher in WCAHF group than in LCAF and CF groups.One patient died of radiation pneumonitis in WCAHF group. Conclusions Whole course accelerated hyperfractionation radiotherapy can significantly improve the local control rates and the 1-and 2-year survival rates of esophageal cancer,but can also increase the toxicity.
6.Experimental study of IRAK-4 expression in hypoxic neurons of rats
Jiangtao LI ; Shengli HU ; Jun HU ; Peng DU ; Hua FENG ; Yunfeng YANG ; Linjie WEI ; Gang ZHU
Chinese Journal of Trauma 2010;26(7):648-652
Objective To investigate the dynamic changes of interleukin-1 receptor-associated kinases (IRAK-4) in hypoxic neurons and explore their role in regulation of inflammatory reaction. Methods The B35 cells exposed to hypoxia of 3% O2,5% CO2 and 92% N2 were cultured for 1,3,6, 12,24,48,72 and 96 hours respectively. Then, mRNA and protein expressions of IRAK-4 were detected by RT-PCR and Western blot, the expression of IRAK-4 in the cells were observed by laser scanning con-focal microscope (LSCM), and the concentration of IL-6 was measured by ELISA method. Results After hypoxia, the mRNA and protein expressions of IRAK-4 were increased at one hour, reached the peak at six hours (P<0.05), kept at a high level at 12 hours (P<0.05) , but decreased gradually to the normal oxygen level at 24 hours (P < 0.05) and to below the normal oxygen level at 48 hours (P < 0.05). Immunofluorescence results showed that the fluorescence intensity of IRAK-4 was gradually increased with time. The changes of IL-6 in the supernatant were positively correlated with protein expression of IRAK-4 (r =0.84, P < 0.05 ). Conclusions Hypoxia can increase the expression of IRAK-4 at transcription and translation levels in a certain period of time, which may participate in down-stream inflammatory reaction and lead to increase of IL-6 expression.
7.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,
9.Optimization of spraying dry technology of Biqiu ranules ethanol extract by Box-Behnken response surface method.
Yan-jun ZHANG ; Li-li LIU ; Jun-hua HU ; Yun WU ; En-xiang CHAO ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(18):3585-3589
With inlet temperature, specific gravity, feeding speed as independent variables, the comprehensive evaluating indexes of content of schisandrin and arctiin as dependent variable, the experimental data were fitted to a second order polynomial equation. Based on establishing the mathematical relationship between the comprehensive evaluating indexes and respective variables, Box-Benhnken central composite test and response surface analysis method was employed to optimize the spray drying technology of Biqiu granules ethanol extract. The optimal drying parameter was as follows: the inlet temperature was 175 degrees C, the specific gravity was 1.10, feeding speed was 32 r x min(-1). Under these conditions, the comprehensive evaluating indexes of spraying dry processes was 92.68, which was close to the model prediction. The spraying dry technology of Biqiu granules ethanol extract optimized by response surface methodology was accurate and feasible, which provided theoretical experiment basis for the industrialization production.
Chemistry, Pharmaceutical
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methods
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Ethanol
10.Optimization of one-step pelletization technology of Biqiu granules by Plackett-Burman design and Box-Behnken response surface methodology.
Yan-jun ZHANG ; Li-li LIU ; Jun-hua HU ; Yun WU ; En-xiang CHAO ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(22):4406-4410
First with the qualified rate of granules as the evaluation index, significant influencing factors were firstly screened by Plackett-Burman design. Then, with the qualified rate and moisture content as the evaluation indexes, significant factors that affect one-step pelletization technology were further optimized by Box-Behnken design; experimental data were imitated by multiple regression and second-order polynomial equation; and response surface method was used for predictive analysis of optimal technology. The best conditions were as follows: inlet air temperature of 85 degrees C, sample introduction speed of 33 r x min(-1), density of concrete 1. 10. One-step pelletization technology of Biqiu granules by Plackett-Burman design and Box-Behnken response surface methodology was stable and feasible with good predictability, which provided reliable basis for the industrialized production of Biqiu granules.
Chemistry, Pharmaceutical
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methods
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Drugs, Chinese Herbal
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chemistry
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Temperature