1.Establishment and evaluation of a BALBc mouse model of Burkholderia pseudomallei via nasal infection
XIA Fei ; QU Lei ; HU Hai-tao ; YANG Fang
China Tropical Medicine 2022;22(10):923-
Abstract: Objective To establish an animal model of BALB/c mice infected with Burkholderia pseudomallei through the nose (inhalation route), provides a reliable animal model for the follow-up studies on the virulence of melioidosis and the pathogenesis of acute melioidosis. Methods The experiment was carried out through infecting with Burkholderia pseudomallei through the nose (inhalation route). The pathophysiological response, visceral pathological damage and bacterial colonization of the mice infected with Burkholderia pseudomallei were observed by gross anatomy, histopathology and tissue homogenate count, and the biological characteristics of the mouse model of acute melioidosis were analyzed accordingly. Then we compared the physiological responses in BALB/c mice between the Burkholderia pseudomallei and non-pathogenic Burkholderia thailandensis. Results In the model of acute nasal infection with Burkholderia thailandensis, most death happened between the 3rd to 5th day after infection, 3×105-3×106 CFU was the suitable dose for acute fatal melioidosis model of BALB/c mice, and the medium lethal dose was about 3×104-3×105 CFU. Both gross anatomy and tissue HE staining showed that abscesses or necrosis were found in the lung, spleen and liver, especially in the spleen and lung, which was positively correlated with the challenge dose. Viable bacteria was isolated from the blood, lung, spleen and liver of Burkholderia pseudomallei-infected mice, and the bacteria account colonization was related to tissue specificity. The concentration of live bacteria isolated from in the blood was the highest [Log2 value: (10.28±0.34) CFU/mL], and the organ with the maximum quantity of bacteria was the lung [Log2 value: (7.54±2.11) CFU/total organ]. It has been reported that the biological effects of Burkholderia pseudomallei and its homologous non-pathogenic Burkholderia thailandensis were similar at the cellular level, like multi-nuclear giant cell formation and active intracellular replication, while it is still unclarrified in the differences of virulence in mice. In this study, it was proved that Burkholderia thailandensis was not fatal to mice even at a high dose (8×107CFU), or detected from mice infected with it via nasal. Conclusion We successfully established a reliable BALB/c mouse model (acute lethal model) of melioidosis via nasal infection, described its biological characteristics, and identified the different biological responses between Burkholderia pseudomallei and its homologous non-pathogenic Burkholderia thailandensis in mice.
2.Identification and Function of Acid-sensing Ion Channels in RAW 264.7 Macrophage Cells
Lan NI ; Peng FANG ; Zhuang-Li HU ; Hai-Yun ZHOU ; Jian-Guo CHEN ; Fang WANG ; You JIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2018;38(3):436-442
Activation of acid-sensing ion channels (ASICs) plays an important role in neuroinflammation.Macrophage recruitment to the sites of inflammation is an essential step in host defense.ASIC 1 and ASIC3 have been reported to mediate the endocytosis and maturation of bone marrow derived macrophages.However,the expression and inflammation-related functions of ASICs in RAW 264.7 cells,another common macrophage,are still elusive.In the present study,we first demonstrated the presence of ASIC 1,ASIC2a and ASIC3 in RAW 264.7 macrophage cell line by using reverse transcriptase polymerase chain reaction (RT-PCR),Western blotting and immunofluorescence experiments.The non-specific ASICs inhibitor amiloride and specific homomeric ASIC 1 a blocker PcTx 1 reduced the production of iNOS and COX-2 by LPS-induced activating RAW 264.7 cells.Furthermore,not only amiloride but also PcTx 1 inhibited the migration and LPS-induced apoptosis of RAW 264.7 cells.Taken together,our findings suggest that ASICs promote the inflammatory response and apoptosis of RAW 264.7 cells,and ASICs may serve as a potential novel target for immunological disease therapy.
3.Dietary patterns and its influencing factors among freshmen students in college
Su-Fang WANG ; Min MU ; Yan ZHAO ; Hu-Zhong LI ; Yan-Fu FANG ; Hai-Lin WANG ; Li LI ; Chuan-Lai HU
Chinese Journal of Epidemiology 2011;32(9):869-872
Objective To investigate the dietary pattern in college freshmen students and to analyze the influencing factors on their dietary patterns. Methods A questionnaire survey on situation of dietary pattern and influencing factors was conducted among 1319 freshmen students.Results Four major dietary patterns were noticed and they were: Ⅰ , high consumption in hamburger,fried food, nuts, biscuit, chocolate, cola, coffee, sugars, Ⅱ, high consumption in pork, mutton, beef,poultry meat, animal liver, Ⅲ, high consumption in fresh fruits, eggs, fish and shrimps, kelp laver and sea fish, milk and dairy products, beans and bean products, Ⅳ, high consumption in rice and grain,fresh fruits, fresh vegetables, pork. Risk factors on dietary pattern were presented as follows: ( 1 )boys: having the food pattern Ⅰ and Ⅱ showed a strong positive association with the place where they live (OR= 1.67, 95%CI: 0.87-3.19; OR= 1.51,95%CI: 0.79-2.88), eating place (OR=1.63,95%CI: 1.O3-2.59; OR= 1.83, 95%CI: 1.04-3.23), level of mother' s education (OR=2.52,95%CI: 1.07-5.95; OR=3.38, 95%CI: 1.50-7.63), family income (OR=2.24, 95%CI: 1.30-3.88;OR=3.06, 95% CI: 1.77-5.29) and the status of passive smoking (OR= 1.80, 95%CI: 0.70-4.59;OR=1.83, 95%CI: 0.75-4.45). Inverse correlations was found on the level of mother's education (OR=0.56,95%CI: 0.17-1.79). The food pattern Ⅳ showed a strong positive association with place of eating(OR= 1.83,95%CI: 1.04-3.23) but having an inverse correlation with the level of mother's education (OR=0.56, 95%CI: 0.17-1.79). (2)girls when compared with boys, the food pattern Ⅰ showed minor association with the places of living and eating; while the food pattern Ⅱ and Ⅲ had minor association with the status of passive smoking. Conclusion Socio-demographic factors and lifestyle had influenced on the dietary patterns among college freshmen students who should be guided to have a reasonable, balanced diet in the college.
4.Genome-wide association study of high altitude pulmonary edema.
Rui-Feng DUAN ; Wei LIU ; Chao-Liang LONG ; Yan-Fang ZHANG ; Wen-Yu CUI ; Yin-Hu WANG ; Hai WANG
Chinese Journal of Applied Physiology 2014;30(2):101-105
OBJECTIVEHigh altitue pulmonary edema (HAPE) impacts seriously people's health at high altitude. Screening of susceptibility genes for HAPE will be used for the evaluation and protection of susceptible people.
METHODSWe performed a genome-wide association study (GWAS) using Affymetrix SNP array 6.0 in 23 HAPE patients and 17 healthy controls. GO and Pathway analysis softwares were used to analyze and draw gene network.
RESULTSThirty-nine SNPs were found to be significantly different between case and control groups (P < 10(-4)). GO and Pathway analysis of 27 genes around the 39 SNPs indicated that these genes mainly participate in the regulating of cell proliferation, regulation of nitrogen compound metabolic process and G-protein coupled receptor protein signaling pathway and so on.
CONCLUSIONIt suggests that these SNPs and genes found in this study may be associated with the susceptibility of HAPE.
Adult ; Altitude Sickness ; genetics ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Humans ; Hypertension, Pulmonary ; genetics ; Polymorphism, Single Nucleotide ; Young Adult
5.Determination of 27 elements in Maca nationality's medicine by microwave digestion ICP-MS.
Gui-fang YU ; Hai-jie ZHONG ; Jun-hua HU ; Jing WANG ; Wen-zhe HUANG ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(23):4545-4551
An analysis method has been established to test 27 elements (Li, Be, B, Mg, Al, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Sr, Mo, Cd, Sn, Sb, Ba, La, Hg, Pb, Bi) in Maca nationality's medicine with microwave digestion-ICP-MS. Sample solutions were analyzed by ICP-MS after microwave digestion, and the contents of elements were calculated according to their calibration curves, and internal standard method was adopted to reduce matrix effect and other interference effects. The experimental results showed that the linear relations of all the elements were very good; the correlation coefficient (r) was 0.9994-1.0000 (Hg was 0.9982) ; the limits of detection were 0.003-2.662 microg x L(-1); the relative standard deviations for all elements of reproducibility were lower than 5% (except the individual elements); the recovery rate were 78.5%-123.7% with RSD lower than 5% ( except the individual elements). The analytical results of standard material showed acceptable agreement with the certified values. This method was applicable to determinate the contents of multi-elements in Maca which had a high sensitivity, good specificity and good repeatability, and provide basis for the quality control of Maca.
Lepidium
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chemistry
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Mass Spectrometry
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methods
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Microwaves
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Reproducibility of Results
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Trace Elements
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chemistry
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isolation & purification
6.Expression of GIRK4 gene in kidney tissues of obese rat.
Yong-an KANG ; Yan-rong HU ; Li GAO ; Hai YANG ; Nan-fang LI
Acta Academiae Medicinae Sinicae 2013;35(1):36-39
OBJECTIVETo investigate the expression of GIRK4 gene in the kidney tissues of obese rats.
METHODSObese rat models were established using diet-induced method. The GIRK4 protein expression in kidney tissues was determined in 20 obese rats and 10 normal rats using Western blot analysis.
RESULTSThe relative expression level of GIRK4 protein in the kidney tissues of obese rat (1.75±0.42) was significantly lower than that in normal rats (3.37±0.68, P<0.05).
CONCLUSIONGIRK4 has a low protein expression in the kidney tissues of obese rat.
Animals ; Female ; Gene Expression ; Kidney ; metabolism ; Male ; Obesity ; genetics ; metabolism ; Potassium Channels, Inwardly Rectifying ; genetics ; metabolism ; Rats
7.Determination of eleutheroside B in antifatigue fraction of Acanthopanax senticosus by HPLC.
Chen LI ; Xiao-yan WANG ; Xu-wei HU ; Hai-tao FANG ; Shan-yi QIAO
China Journal of Chinese Materia Medica 2008;33(23):2800-2802
OBJECTIVETo develop an HPLC method for the determination of eleutheroside B in As1 (the extract fraction from Acanthopanax senticosus, which has a good effect of antifatigue).
METHODThe antifatigue effect of Asl was evaluated by mice burden swimming testing. One compound was isolated by silica gel, Sephadex LH-20 column chromatography and preparative high performance liquid chromatography. The structure was identified by physicochemical properties and spectral evidences. Eleutheroside B in Asl was determined by HPLC. Chromatographic conditions included Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) and the mobile phase consisting of a mixture of acetonitrile-water-ethanoic acid (10: 90: 0.01). The UV detection wavelength was set at 344 nm.
RESULTAs1 showed an excellent antifatigue activity; One compound was isolated from As1 and its structure was identified as Eleutheroside B; The calibration cure was linear in the range of 0.104-20.8 microg (r = 0.9999), the average recovery was 97.68%, RSD 1.4% (n=6).
CONCLUSIONThis HPLC method is simple,accurate and reliable.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eleutherococcus ; chemistry ; Fatigue ; drug therapy ; physiopathology ; Glucosides ; analysis ; Humans ; Male ; Mice ; Phenylpropionates ; analysis ; Swimming
8.Research progress on effect factors of secondary metabolites content in callus.
Hui-Yong FANG ; Hong ZHU ; Hai-Mai DING ; Hua-Rui HAN ; Xiao-Lan LIU ; Li-Jing HAO ; Min-Hu LI
China Journal of Chinese Materia Medica 2014;39(15):2846-2850
Secondary metabolites are the result of that plant interaction with biological and non-biological factors in the long-term evolution process, and play an important role in plant growth, development and physiology. The effective components of medicinal plant are usually the secondary metabolites in plant cells, and the synthesis of them are affected by a variety of factors, such as environmental impact. Acquirement of the secondary metabolites via callus culture has the advantage of low cost and less environmental impact. The synthesis and accumulation of medicinal plant secondary metabolites are not only controlled by light, temperature and pH, but also infected by germplasm, plant growth regulator and elicitor. This article presents a review of the influencing factors, and provides a basis for further study and development.
Light
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Plant Growth Regulators
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metabolism
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Plant Physiological Phenomena
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radiation effects
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Plants
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metabolism
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radiation effects
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Temperature
9.Mini-open repair for the treatment of acute closed achilles tendon ruptures.
Tao CHEN ; Xin ZHENG ; Hong-bin AN ; Yang HUANG ; Fang-hu CHEN ; Jian-wei RUAN ; Hai-bao WANG
China Journal of Orthopaedics and Traumatology 2015;28(7):654-656
OBJECTIVETo evaluate the clinical efficacy of mini-open repair for the treatment of acute closed achilles tendon ruptures.
METHODSFrom April 2012 and October 2013,14 patients (14 feet) with acute closed achilles tendon ruptures were treated in our department. They were 9 males and 5 females, with an average age of 30.5 years old (ranged, 25 to 49 years old). The interval between injury and operation ranged from 1 to 13 days (8 days on average). A longitudinal incision approximately 1.5 to 2.0 cm in length was made around the ruptured achilles tendon for mini-open repair after insertion of oval clamp. Postoperative rehabilitation was carried out.
RESULTSThe wounds healed at the first stage except 2 cases with slow recovery. All the patients were followed up for 6 to 24 months, with an average of 11 months. According to the ankle-hindfoot scoring system of American Orthopaedic Foot & Ankle Society (AOFAS),the score was 92.71 ± 6.58 (82 to 100).
CONCLUSIONThe surgical treatment of acute achilles tendon rupture with mini-open repair has advantages of little invasion, a low rate of incision problems, quick function recovery, and simple operation, and it is suitable for primary hospital.
Achilles Tendon ; injuries ; physiopathology ; surgery ; Adult ; Female ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Tendon Injuries ; physiopathology ; surgery ; Treatment Outcome ; Wound Healing