1.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.
2.The reliability and validity of the Chinese version of the upper extremity functional index
Xiling XIAO ; Zhaohui YANG ; Xiaoxuan XIA ; Shuang WANG
Chinese Journal of Physical Medicine and Rehabilitation 2012;(12):903-906
Objective To adapt the upper extremity functional index (UEFI) to Chinese and assess the reliability and validity of the Chinese version.Methods The index items were translated into Chinese,back translated into English,and revised by experts in rehabilitation.Twenty-five patients with upper extremity trauma were then recruited to complete two UEFI questionnaires on two occasions 1 week apart as a field test.They also completed a disability of arm,shoulder and hand (DASH) questionnaire on the first occasion.Results Some adjustments were made to the translations based on cultural and linguistic practices.Cronbach's alpha for the final version was 0.93 and the total inter-class correlation coefficient (ICC) was 0.97.The ICCs of the individual items ranged from 0.56 to 0.94.The Pearson correlation coefficient between the UEFI and DASH questionnaires was-0.86.Conclusions The Chinese version of the UEFI scale is reliable and valid.It can be used for study and in the clinical evaluation of upper extremity function.
3.Clinical observation on the neurotransmitters regulation in patients of insomnia differentiated as yang deficiency pattern treated with warm acupuncture and auricular point sticking therapy.
Jin-Liang YANG ; Rong ZHANG ; Lei DU ; Yun-Shuang YANG ; Xiao-Chen LIU
Chinese Acupuncture & Moxibustion 2014;34(12):1165-1168
OBJECTIVETo evaluate the effect mechanism of warm acupuncture combined with auricular point sticking therapy and its efficacy on insomnia by monitoring the level of brain neurotransmitters in the insomnia patients.
METHODSOne hundred and thirty patients with insomnia were randomized into an observation group and a control group, 65 cases in each one. In the observation group, based on the treating principle of warming yang and benefiting qi, acupuncture was applied to Xinshu (BL 15), Pishu (BL 20), Shenshu (BL 23), Yaoyangguan (GV 3), Baihui (GV 20), Neiguan (PC 6) and Shenmen (HT 7). Warm acupuncture was supplemented at Xinshu (BL 15), Pishu (BL 20), Shenshu (BL 23) and Yaoyangguan (GV 3). The treatment was given once every day. In the control group, estazolam tablets, 0. 5 to 1 mg were prescribed for oral administration, 30 min before sleep at night. The treatment of 14 days was taken as one session and 2 sessions were required in the two groups. The encephal of luctuograph technology was used to observe the sleep quality and brain neurotransmitters before and after treatment in the two groups and the efficacy was compared between the two groups.
RESULTSThe total effective rate was 83.1% (54/65) in the observation group and 87.7% (57/65) in the control group. The efficacy was similar between the two groups (P > 0.05). In the observation group, after treatment, 5-HT and GABA/Glu were all increased compared with those before treatment (P < 0.05, P < 0.01) and norepinephrine (NE) was reduced compared with that before treatment (P < 0.05). The level of each index did not change significantly before and after treatment in the control group (all P > 0.05). The regulations of 5-HT, GABA/Glu in the observation group were superior to those in the control group (all P < 0.05).
CONCLUSIONThe combined therapy of warm acupuncture and auricular point sticking method for warming yang and benefiting qi effectively improves brain neurotransmitters and essentially improves sleep quality of the patients with insomnia differentiated as yang deficiency pattern.
Acupuncture Points ; Acupuncture Therapy ; Acupuncture, Ear ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neurotransmitter Agents ; metabolism ; Serotonin ; metabolism ; Sleep Initiation and Maintenance Disorders ; metabolism ; therapy ; Treatment Outcome ; Yang Deficiency ; metabolism ; therapy ; Young Adult ; gamma-Aminobutyric Acid ; metabolism
4.Advances in studies on biotransformation of ginsensides.
Cong-Liang GUO ; Xiu-Ming CUI ; Xiao-Yan YANG ; Shuang WU
China Journal of Chinese Materia Medica 2014;39(20):3899-3904
Ginseng saponins are a type of important active substances in the ginseng genus plants. They have notable pharmacological activities of antineoplastic, neuroprotective, and hepatoprotective activities, which have been drawn more attention to obtain minor ginsenosides by all kinds of methods. In this review, we discussed the latest progress for enrichment of minor ginsenosides by biological transformation of major ginsenosides. At the same time, we have a brief outlook of the research at bioconversion of ginseng saponins.
Bacteria
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metabolism
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Biotransformation
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Drugs, Chinese Herbal
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chemistry
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metabolism
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Ginsenosides
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chemistry
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metabolism
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Panax
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chemistry
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metabolism
5.Expression of hypoxia-inducible factor 2 alpha and vascular endothelial growth factor in chondrocytes of articular cartilages in human osteoarthritis
Feng LIU ; Hao PENG ; Jianlin ZHOU ; Hongsong FANG ; Shuang DENG ; Xiao YANG ; Jinqing WENG
Chinese Journal of Tissue Engineering Research 2015;(2):201-206
BACKGROUND:Studies have found that vascular endothelial growth factor and hypoxia inducible factor are involved in the development process of osteoarthritis, but their correlation is rarely reported. OBJECTIVE:To observe the expression and correlation of hypoxia inducible factor-2α and vascular endothelial growth factor in chondrocytes of articular cartilages in human osteoarthritis. METHODS: Articular cartilage specimens were colected from 50 patients with knee osteoarthritis undergoing total knee joint replacement. According to the joint Kelgren-Lawrance (K-L) X-ray grouping classification standard, there were 18 cases of K-LIII level and 32 cases of K-LIV level. Besides, articular cartilage specimens from 10 patients undergoing amputation for legs tumor or traffic accident served as control group. Hematoxylin-eosin staining, Safranin O-Fast Green staining and Mankin scoring were performed to observe and evaluate the histological characteristics of articular cartilages of each group, immunohistochemical staining was conducted to detect the expression of hypoxia inducible factor-2α and vascular endothelial growth factor in chondrocytes of articular cartilages, and their correlations were analyzed. RESULTS AND CONCLUSION:The Mankin score of K-LIV group was significantly higher than those of K-LIII group and control group. Immunohistochemical staining revealed that the number of chondrocytes with positive expression of hypoxia inducible factor-2α or vascular endothelial growth factor in K-LIV group was significantly higher than that in K-LIII group and control group (P < 0.05). The expression of hypoxia inducible factor-2α and vascular endothelial growth factor increased in chondrocytes of articular cartilages of osteoarthritis patients, and to up-regulate the expression of vascular endothelial growth factor may be the regulatory mechanism of hypoxia inducible factor-2αinthe pathogenesis of osteoarthritis.
6.Expression of Trem-1 and its significance in psoriasis vulgaris
Chen TU ; Shuang WANG ; Yang ZHANG ; Jianwen REN ; Xiaopeng WANG ; Shengxiang XIAO
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;37(3):428-431
Objective To study the expression of triggering receptor expressed on myeloid cells-1 (Trem-1)in psoriatic vulgaris and normal skin tissues and blood,and to explore the potential pathogenesis of psoriasis.Methods Immunohistochemistry and Real-time PCR were used to detect the expression of Trem-1 in the blood and tissues of normal skin and psoriasis.Results The positive expression rate of Trem-1 in psoriatic lesion was significantly higher than normal tissue.Trem-1 was expressed in the whole epidermis,with a significant difference (P<0.05). The mRNA expression of Trem-1 was significantly higher in psoriatic skin tissues and blood than in normal skin tissues and blood (P<0.05).Moreover,the mRNA expression of Trem-1 was positively correlated with PASI (P<0.05).Conclusion Abnormal expression of Trem-1 might be related to the pathogenesis of psoriasis.Trem-1 will cure psoriasis vulgaris as the potential therapeutic target.
7.Characteristic Analysis of Cooperation Hydrogen Production Using Rhodopseudomonas sp. DT and Enterobacter Aerogenes
Xiao-Rong ZHANG ; Shuang-Jiao GONG ; Hui-Min LIAO ; Dong-Mei YANG ; Yi-Guang CHEN ;
Microbiology 2008;0(10):-
Cooperation hydrogen production was carried out using Rhodopseudomonas sp. DT and Enterobacter aerogenes. The effects of the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes, culture temperature, and carbon source on the cooperation hydrogen production were investigated. The results suggested that cooperation hydrogen production rate was highly affected by the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes. The mixed bacteria of Rhodopseudomonas sp. DT and E. aerogenes with 1:1 initial ratio benefited to the cooperation hydrogen production, which led the hydrogen production rate and duration of gas production to 3.1 mol H2/mol glucose and 81 h, respectively. The pH dynamics analysis of culture medium further discovered that the pH of the mixed bacteria with 1:1 initial ratio changed from 6 to 7 smaller than other conditions, which was probably fitted to produce hydrogen. Furthermore, the mixed bacteria with 1:1 initial ratio had the higher hydrogen production efficiency at temperatures of 28?C and 37?C than at 20?C, and without any hydrogen production at temperature of 50?C. The carbon sources of glucose, succinate acid, malic acid could be used to produce hydrogen by the mixed bacteria. Even the soluble starch, unused by Rhodopseudomonas sp. DT, was also decomposed by the mixed bacteria to produce hydrogen with the conversion efficiency of 8.22%. The glucose was the optimal carbon resource, and the conversion efficiency could reach to 36.11%. The results, further, implied that the cooperation hydrogen production could enlarge the use of the carbon sources.
8.Experimental study and clinical application of anti-adhesion treatment and high-strength suture technique on the treatment of extensor tendon rupture
Dawei JIANG ; Quanzhang JIA ; Mingxiao HOU ; Shuang XU ; Guang XIAO ; Yang CHEN ; Dongjun LI
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
Objective To investigate the effects of anti-adhesion treatment and high-strength suture technique on the treatment of extensor tendon rupture by animal experiments and clinical application.Methods Twenty-eight leg-born chickens were randomly divided into four groups(7 each).Double cross suture was applied in group A,while double cross suture combined with sodium hyaluronate spraying in group B,cross-finger-like micro-braided suture in group C,and cross-finger-like micro-braided suture combined with sodium hyaluronate spraying in group D.The animals were sacrificed 6 weeks after operation,morphological,histological and biomechanics were observed and compared among the groups.One hundred and sixteen patients were treated with the surgical method in group D(89 males,27 females,aged 20-55 with an average of 36 years;73 with extensor tendon rupture,38 with strain/chalasis,5 with firearm injuries;56 on back of hand,48 on central slip,12 on lateral fixing chorda;82 with one-stage operation,and 34 with second-stage operation),and then followed-up for 2-5 years to observe the therapeutic effects.Results The repaired tendons in group D was in good contour,most tendon cells arranged regularly in bunches.The maximum load was significantly higher in group D(70.9?5.7N) than in group A(48.4?5.7N),Group B(51.3?3.2N) and Group C(68.3?2.8N,P
9.Subcellular localization of macrophage migration inhibitory factor and its diverse biological functions in tumors
Huan ZHANG ; Chen YANG ; Shuang SHANG ; Xiao-xi LV ; Fang HUA
Acta Pharmaceutica Sinica 2023;58(8):2085-2097
Macrophage migration inhibitory factor (MIF) is an enzyme-active pleiotropic cytokine that is expressed in various immune cells and tumor cells. MIF plays diverse roles in inflammation and tumor progression. It acts as a cytokine involved in immune response and inflammatory lesions. Additionally, MIF is closely associated with tumor proliferation, metastasis, and other tumor hallmarks, exerting a multifaceted influence on tumor occurrence and progression. MIF not only functions by being secreted into the extracellular space as a cytokine but can also be localized within the cytoplasm and nucleus, exhibiting diverse biological functions. As MIF in promoting tumor progression becomes increasingly recognized, MIF-based therapeutic strategies have become a hot research topic in oncology. Here, we provide a comprehensive review of MIF with different subcellular localization about their pro-tumoral functions. A better understanding of MIF in tumor biology will bring broader perspectives for the development of novel MIF targeting strategies and give promising direction for future tumor treatments.
10.Craniofacial characteristics of the Class II division 1 malocclusion patients with mouth-breating: a posteroanterior.
Xiao-jun LIU ; Shuang YAO ; Shuang YANG ; Ping YANG ; Qing XU ; Bing WANG ; Wen-hong WANG
West China Journal of Stomatology 2005;23(3):214-216
OBJECTIVETo evaluate the craniofacial characteristics of the Class II malocclusion patients with mouth-breating by posteroanterior cephalometry.
METHODSTo measure craniofacial width of the 12 Class II malocclusion patients with mouth-breathing, and to compared these measures with corresponding measures in a group of normal children.
RESULTSThe width of the maxillary base bone (J-J) was less than that in normal children significantly (P < 0.01). The mouth-breathing children's upper and lower arch width (at first molar and cuspid) were comparatively narrower, and lateronasal width (Lap-Lap) was narrower too.
CONCLUSIONMouth breathing may lead to craniofacial morphological abnormal development in craniofacial transverse structures.
Adolescent ; Cephalometry ; Child ; Cuspid ; Dental Arch ; Face ; Humans ; Male ; Malocclusion ; Malocclusion, Angle Class II ; Maxilla ; Molar