1.Effects of amyloid-beta 25-35 on expression of synapse-associated proteins in PC12 neurons Effects of amyloid-beta 25-35 on expression of synapse-associated proteins in PC12 neurons
Shuang ZHANG ; Xinyan HUANG ; Shuang LIU ; Yanjun LI ; Jincheng ZHAO
Chinese Journal of Tissue Engineering Research 2016;(2):224-229
BACKGROUND:An amyloid-beta (Aβ) aggregation in the brain can induce nerve cel apoptosis, loss of synapses and functional damage. However, there is stil no effective intervention. Improving the synaptic plasticity provides an important direction for the treatment of early Alzheimer’s disease. OBJECTIVE: To screen the best model of Alzheimer’s disease and to explore the expression of synapse-associated proteins in Aβ25-35-injured PC12 neurons. METHODS:PC12 cels were induced by 50 μg/L nerve growth factor to differentiate into neuronal-like cels. Then, these cels were treated with Aβ25-35 at different concentrations. Consequently, cel survival rate was detected using cel counting kit-8; neurogranin and neuregulin immunofluorescence stainings were used to observe morphological changes of model cels; western blot used to detect the expression level of neurogranin, calmodulin kinase II, postsynaptic density-95 proteins. RESULTS AND CONCLUSION:Over time, the survival rate of PC12 neurons induced by Aβ25-35 was decreased in a dose-dependent manner. Shortened synaptic length, neuronal atrophy and sparsely interconnected neurons were visible. Expression levels of neurogranin, calmodulin kinase II and postsynaptic density-95 proteins were al down-regulated. These findings indicate that to screen the cel model of Alzheimer’s disease, the optimal concentration and interventional time of Aβ25-35are 10 μmol/L and 48 hours, respectively.
2.Comparison between Radiofrequency Coblation and CO2 Laser for the Treatment of Early Glottic Carcinoma
Yu SHUANG ; Chao LI ; Yongwang HUANG ; Shuang WU
Journal of Audiology and Speech Pathology 2015;(4):372-376
Objective To comparison curative effects between radiofrequency coblation and CO 2 laser in treat‐ment of early glottic carcinoma .Methods A total of 104 patients with early glottic cancer (Tis ,T1a ,T1b ) were treated in our department from October 2007 to December 2012 .Among them 64 cases underwent radiofrequency ablation (radiofrequency group) while 40 cases underwent CO2 laser resection (CO2 laser group) .The operation time and the postoperative pain VAS scores were recorded .All of the patients were examined by electronic laryngoscopy and a‐coustic analysis in 1 week ,1 month ,3 months after surgery .Results The mean operation time of radiofrequency group was 8 .75 ± 1 .62(min)and CO2 laser group was 11 .82 ± 1 .51(min) .The radiofrequency group was better than CO2 laser group(P < 0 .05) .The differences of postoperative pain VAS score was not statistically significant be ‐tween radiofrequency group(2 .79 ± 0 .47) and CO2 laser group (2 .96 ± 0 .64)(P> 0 .05) .The mucosal recovery of radiofrequency group in 1 month after surgery was better than the CO2 laser group (χ2 = 13 .842 ,P< 0 .05) .Subjects in radiofrequency group and CO2 laser group showed statistical significance and linear trends in jitter ,shimmer ,HNR over time by univariate test of repeated measurement ANOVA (P < 0 .05) .The differences of acoustic parameters (jitter ,shimmer ,HNR)between radiofrequency group and CO 2 laser group were statistically significant by multiva‐riate test of repeated measurement ANOVA (P< 0 .05) .The differences of 3 - years survival rate ,local recurrence rate ,recurrence rate with anterior commissure involved and postoperative adhesion rate with anterior commissure in ‐volved were not statistically significant between radiofrequency group and CO 2 laser group (P > 0 .05) .All of the patients had not underwent tracheotomy .They did not have cough ,dyspnea ,severe pain ,hemoptysis ,serious compli‐cations after surgery .Conclusion Both radiofrequency coblation and CO 2 laser are safe and effective surgical meth‐ods in the treatment of early glottic carcinoma .The operation time of radiofrequency coblation is shorter than CO 2 laser surgery .The mucosal recovery in 1 month after surgery and acoustic recovery of radiofrequenay coblation are all better than that of the CO2 laser surgery .
3.Content Determination and Quality Evaluation of Gallic Acid in Gentiana Rigescens from Different Areas in Guizhou Province
Shuang HAN ; Xianrong XI ; Ping HUANG
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(05):-
Objective To determine the content of gallic acid and evaluate the quality of Galla chinensis of different habitats in Guizhou Province. Methods HPLC method was used to determine the content of Gallic acid on Alltima C18 column (4.6 mm?250 mm, 5 ?m) with the mobile phase of methanol-water-phosphoric acid (15∶85∶0.085). The flow rate was 1.0 mL/min and the detective wave length was 270 nm. Results The contents of Gallic acid in Galla chinensis of 10 different habitats in Guizhou Province were 51.43%~ 60.25%. Conclusion The quality of Galla Chinensis in different habitats in Guizhou Province are up to the Chinese Pharmacopoeia, and the individual varibility is significant.
4.Advances of Particles/Cells Magnetic Manipulation in Microfluidic Chips
Shuang HUANG ; Yongqing HE ; Feng JIAO
Chinese Journal of Analytical Chemistry 2017;45(8):1238-1247
Magnetic manipulation of particles/cells in microfluidic chips is a promising research field.This paper stated the operation mechanism and the main means of manipulation, including separation, concentration, capture, arrangement and assembly.Especially, the concept of particles/cells separation was emphasized with different criteria, like sizes, shapes, and magnetic properties.In addition, the effects of the channel geometry, the intensity and distribution of the magnetic field, and the types of magnetic liquid (paramagnetic salt solutions and ferrofluids) on the performance of the magnetic manipulation were also compared.The prospective to the prospect of magnetic manipulation about particles/cells in microfluidic chip was also depicted.
5.The effect of interferon ? with different structures on ACHN tumor suppression
Haixiao HUANG ; Shuang XING ; Ming LI
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
Objective To evaluate the anti-tumor activity of interferon ? with different structures including pegylated interferon ?-1b (PEG IFN ?-1b), pegylated interferon ?-2b (PEG IFN ?-2b) and natural interferon ? (IFN ?) in male BALB/C nude mice with human renal adencarcinoma (ACHN) tumor. Methods Athymic BALB/C nude mice were received a subcutaneous injection of ACHN cells (2?106/0.2ml) on the rear flank. After four weeks, the size of tumor reached a mean volume of 50-100 mm3, and the interferon was administrated. The mice with tumor were divided into four groups, and were administrated with placebo (0.2ml/mouse thrice weekly), IFN ?-1b (50?g/mouse thrice weekly), PEG IFN ?-1b (150?g/mouse once weekly) and PEG IFN?-2b (150?g/mouse once every week), respectively, for five weeks. The tumor volumes were measured weekly prior to treatments until the end of the fifth week. All mice in each group were sacrificed to measure the weight of the residual tumor. The anti-tumor activity of IFN ? in vivo was evaluated by studying their ability to reduce the volume and weight of tumor. Results IFN ? may significantly reduce the volume and weight of ACHN tumor in mice. The average tumor weight was 0.422g in placebo group, while 0.263g, 0.235g and 0.219g in the groups treated with PEG IFN ?-1b, IFN ?-1b and PEG IFN ?-2b, respectively. No significant difference was found among the three groups treated with IFN ?. Moreover, the volume of tumor in three groups treated with IFN ? was significant reduced than that in placebo group. Conclusion IFN ? with different structures have showed the similar ability of ACHN tumor suppression at the dosage of 150?g per week.
6.Inhibitory effects of Interferon alfa-1b on human liver cancer cells in vivo
Shuang XING ; Hongling OU ; Haixiao HUANG
Medical Journal of Chinese People's Liberation Army 2001;0(07):-
Objective To compare the effects of interferon alfa-1b(IFN?-1b) and pegylated IFN?-1b(PEG IFN?-1b) on inhibiting growth of human liver cancer cells in vivo.Methods Human liver cancer cells(HepG-2,2?l06/0.2ml) cultured in log phase were subcutaneously injected into the right rear flank of male BALB/C nude mice to establish the tumor model.After HepG-2 injection,the tumor growth was followed up for 14 days.The mice with HepG-2 tumor were randomly divided into placebo(n=7),PEG IFN?-1b(n=8) and IFN?-1b(n=8) groups.Mice received injection of placebo 0.2ml/mouse each time(three times a week),PEG IFN?-1b 150?g/mouse per injection(once a week) and IFN?-1b 50?g/mouse each time(three times a week),respectively,according to their grouping,and this regime lasted for five weeks.In order to evaluate anti-tumor activity,animals' body weight was measured every week,and the measurements of tumor volume and animals' body weight were continuously done till the mice were sacrificed at the end of the fifth week.Results PEG IFN?-1b and IFN?-1b induced a significant decrease in tumor volume and the animals' weight,the suppression rates onto tumor growth were 56%(2.190g vs.4.979g) and 42%(2.678g vs.4.979g),respectively.However,no significant difference was found in animals' weight among the three groups.Conclusion HepG-2(2?l06/0.2ml) injection into nude mice may induce a typical tumor model.PEG IFN?-1b(150?g/mouse per time,once a week) could significantly inhibit the growth of HepG-2 in vivo,and so does the IFN?-1b(50?g/mouse each time,three times per week).
7.Analysis of critical genes expression of chlorogenic acid and luteolin biosyntheses in Lonicera confusa.
Shuang-Shuang QIN ; Lu-Qi HUANG ; Yuan YUAN ; Li-Ying YU
China Journal of Chinese Materia Medica 2014;39(13):2469-2472
This study analysed the tissue specific expression of critical genes involved in chlorogenic acid and luteolin biosynthesis, for exploiting the molecular mechanism of components biosynthesis in Lonicera confusa. Expression of PAL, 4CL, C4H, CHS, CHI, FNS and HQT gene families of chlorogenic acid and luteolin biosynthesis-related genes in buds and leaves of L. confusa were analyed by Real-time PCR. Expressions of PAL1, C4H1, 4CL1, CHS1, CHI3 and HQT2 in buds were lower than that in leaves, and expressions of PAL3, 4CL2, CHI2 and FNS2 in buds were higher than that in leaves. The results indicated that that PAL3 and 4CL2 may be associated with accumulation of chlorogenic acid, and the expression patterns of PAL1, CHS1, CHI3 and HQT2 in buds and leaves of L. confusa were different with L. japonica. This study provided some theoretical basis for the further research on genetic mechanism of active components differences in L. confusa and L. japonica.
Biosynthetic Pathways
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Chlorogenic Acid
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metabolism
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Gene Expression Regulation, Plant
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Lonicera
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genetics
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metabolism
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Luteolin
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biosynthesis
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Multigene Family
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Plant Proteins
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genetics
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metabolism
8.Construction and identification of RNAi lentiviral vector targeting at triggering receptors expressed on myeloid cells-1
Dajiang SONG ; Xiaoyuan HUANG ; Xinghua YANG ; Muzhang XIAO ; Shuang WANG
Journal of Central South University(Medical Sciences) 2009;34(10):970-977
Objective To construct a lentiviral vector of RNA interference (RNAi) of murine triggering receptor expressed on myeloid cells-1 (TREM-1) gene and to explore the effect of TREM-1 on the inflammatory response caused by Bacteroides fragilis. Methods Four target sequences were selected according to murine TREM-1 mRNA sequence, and then 4 pairs of double-strand DNA oligo according to these target sequences and one pair of negative control double-strand DNA oligo were designed and synthesized. These fragments were subcloned into pGCSIL-GFP/Lenti plasmid. After being identified by PCR and sequencing, these plasmids were cotransfected into 293T cells to package lentiviral particles. The lentiviral vector particles were transfected into Raw 264. 7 cells and TREM-1 expression in the transfected cells was assayed by real-time PCR and ELISA. Different concentrations of Bacteroides fragilis lipopolysaccaride (LPS) were administered in the Raw264. 7 cells, and the cells were stimulated with LPS for 12 h. TREM-1 expression was determined by real-time PCR and ELISA at the time points. Results PCR and sequencing confirmed that lentiviral vectors had the correct structure and could express high titer of virus. After being transfected into Raw264. 7 cells, TREM-1 expression was knocked down significantly by all of these lentiviral vectors at both protein and mRNA levels, and the pGCSIL-GFP/Lenti-1 had the most efficient interference. TREM-1 was upregulated in the presence of Bacteroides fragilis LPS, and this increase was partly abrogated in the TREM-1 siRNA-treated cell models of endotoxemia, depending on the sequence. Conclusion The lentivirus RNAi vector of TREM-1 was constructed successfully. The lentivirus RNAi vector of TREM-1 can inhibit the expression of TREM-1 in the murine endotoxemia model caused by Bacteroides fragilis LPS.
9.Effect of electroacupuncture on inducible nitric oxide synthase mRNA expression in hippocampus of the rats with cerebral ischemia-reperfusion injury
Shuang ZHOU ; Jianhua HUANG ; Bangjiang FANG ; Lingsong YUAN
Chinese Journal of Tissue Engineering Research 2005;9(37):138-140
BACKGROUND: Researches indicate that inducible nitric oxide synthase(iNOS) plays an important role in relieving destruction of blood brain barrier and protecting vascular endothelium and brain tissue in cerebral ischemia-reperfusion injury.OBJECTIVE: To observe the effect of Shuigou (DU26), Neiguan (PC6)and Zusanli (ST36) on the expression of inducible nitric oxide synthase messenger ribonucleic acid (iNOS mRNA) in hippocampus of the rats with cerebral ischemia-reperfusion.DESIGN: Randomized controlled study.SETTING: Shanghai University of Traditional Chinese Medicine, Shanghai Institute of Acupuncture and Meridian and Huashan Hospital Affiliated to Fudan University.METHODS: The experiment was completed at the Experimental Animal Center in Shanghai University of Traditional Chinese Medicine, Threegrade Acupuncture Immunological Laboratory of Shanghai Research Institute of Acupuncture and Meridian and Huashan Hospital Affiliated to Fudan University from December 2003 to December 2004. Totally 40 SD rats were randomly divided as normal group, sham operation group, model group and electroacupuncture group with 10 in each group. The hypertension model rats were induced with bi-kidney-bi-clip method, then the focal cerebral ischemia-reperfusion rat model was induced by reversible occlusion for a middle cerebral artery with a thread on the stroke prone renovascular hypertensive model rat. While the rats in sham operation group were operated without thread insertion. Shuigou (Du 26), bilateral Neiguan(PC 6) and bilateral Housanli (Zusanli, ST 36) were located according to"Atlas of Experimental Animal Acupoints" made by Hua's. Shuigou (Du26): 1 mm below nasal apex right on the Labium leporinum, be punctured 1 mm obliquely upward; Neiguan (PC 6): at the lateral palm of the forearm3 mm above wrist joint, between ulna and radius, be punctured 1 mm perpendicularly; Housanli (Zusanli, ST 36): under knee joint, 5 mm below the capitulum of fibula, be punctured 7 mm perpendicularly. The skin of Shuigou (Du 26) and the root of right ear, Neiguan (PC 6) and Housanli (Zusanli, ST 36) were connected with electroacupuncture Therapeutic Instrument G6805 with the parameters of continuous wave, 120 times/minute in frequency, 1 mA in intensity for 30 minutes. The treatment was applied once immediately after ischemia operation and again 12 hours after reperfusion. Being killed on 24 hours after reperfusion, the hippocampus of the rats were rapidly picked out. The effect of electroacupuncture on the expression of iNOS mRNA in hippocampus of the rats with cerebral ischemia-reperfusion was detected with fluorescence quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR).MAIN OUTCOME MEASURES: The expression of iNOS mRNA in hippocampus.RESULTS: All 40 SD rats entered the final analysis. Expression of iNOS mRNA in hippocampus: [1] That in model group was significant higher than that in electroacupuncture group {[(4.85±1.29) ×1 000, (3.19±1.38) ×1 000] copy, (t=2.77, P < 0.05)}; [2] That in model group was significant higher than that in sham operation group {[(4.85±1.29) ×1 000, (4.93±2.17)×10] copy, (t=97.38, P < 0.01)}; [3] That in model group was significant higher than that in normal group {[(4.85±1.29) ×1 000, (3.13±1.68) ×10]copy, (t=11.81, P < 0.01)}.CONCLUSION: Electroacupuncture may significantly inhibit the expression of iNOS mRNA in hippocampus and reduce the synthesis of NO to relieve the cerebral ischemia-reperfusion injury.
10.Law of acupoint selection in acupuncture treatment for insomnia based on data mining method.
Kaiyu HUANG ; Shuang LIANG ; Yueting XU ; Shengfeng LU
Chinese Acupuncture & Moxibustion 2015;35(9):960-963
The modern statistical and data mining technology is used to analyze the law of the acupoint selection in the treatment of insomnia with acupuncture and moxibustion. The clinical literature on the treatment of insomnia with acupuncture and moxibustion in recent 10 years is collected from China National Knowledge Infrastructure (CNKI). Excel software is applied to establish the database of acupuncture prescriptions for insomnia so as to conduct the descriptive analysis, association rule analysis and clustering analysis on the data. In the treatment of insomnia with acupuncture and moxibustion, Shenmen (HT 7) is of the highest frequency and most acupoints were selected from the Governor Vessel. The commonly used acupoints are located on the head, face, neck and nape region. The combination of the local acupoints with the distal ones is predominated. The crossing points among the specific points present the advantage in the treatment. The association: rule analysis indicates the highest, correlation among Taixi (KI 3), Sishencong (EX-HN 1) and Shenmen (HT 7). The clustering analysis results in 6 effective clusters and 10 pairs of key points and summarizes the common law of the acupoint combination for insomnia. All of these provide the reference to the treatment of insomnia with acupuncture and moxibustion.
Acupuncture Points
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Acupuncture Therapy
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China
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Data Mining
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Databases, Factual
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Humans
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Moxibustion
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Sleep Initiation and Maintenance Disorders
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therapy
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Software