1.Culture Methods of the Oligotrophic Marine Microbes
Tian TIAN ; Dong-Mei LI ; Shi-Kun DAI ; Ke-Dong YIN ; Hui-Min SUN ; Xiang LI ;
Microbiology 2008;0(07):-
Molecular methods and fluoroscopic techniques suggest that rich microbial diversity exist in the marine environment, but less than 1% of these microbes can be cultured in the laboratory conditions, and that the cultivable dominant species were even less. This limitation has long been a barrier to the development of environmental microbiology and the utilization of marine resources. In the past decade, novel methods for culture and detection of these uncultured marine microbes have successfully applied to obtain several conventionally-uncultured microbes including those from extreme environments. Those progresses have inspired researchers greatly. Developments in the research of marine microbial resources are an important basis for the study of the micro-world and deserve increasing scientific attention.
2.Effect of andrographolide derivative Yanhuning on in vivo Candida albicans biofilms in rats.
Gao-Xiang SHI ; Yuan-Yuan YAN ; Jing SHAO ; Meng-Xiang ZHANG ; Ke-Qiao LU ; Tian-Ming WANG ; Chang-Zhong WANG
China Journal of Chinese Materia Medica 2014;39(15):2924-2929
OBJECTIVETo investigate the effect of andrographolide derivative Yanhuning (YHN) on Candida albicans biofilms in rats.
METHODThe rat C. albicans biofilms subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2.5 mg x kg (-1)), with the FLC (80 mg x kg(-1)) positive group as the control group. After 7 d, CFU counting and XTT assay were used to evaluate the effect of YHN on C. albicans biofllms in vivo. Scanning electron microscopy (SEM) was applied to observe the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PCR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1, ALS3, HWP1, EAP1 and MP65.
RESULTThe YHN group showed much less CFUs on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SEM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR's results, YHN can down-regulate expressions of ALS1, ALS3, HWP1, EAP1 and MP65.
CONCLUSIONYHN could inhibit C. albicans biofilms in rats.
Animals ; Biofilms ; drug effects ; growth & development ; Candida albicans ; cytology ; drug effects ; physiology ; Catheters ; microbiology ; Cell Adhesion ; drug effects ; Diterpenes ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Rats
3.Effect of Huanglian Jiedu decoction in combination with fluconazole on ergosterol of fluconazole-resistant Candida albicans.
Yuan-yuan YAN ; Tian-ming WANG ; Gao-xiang SHI ; Meng-xiang ZHANG ; Ke-qiao LU ; Jing SHAO ; Chang-zhong WANG
China Journal of Chinese Materia Medica 2015;40(4):727-732
OBJECTIVETo investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) , alone and in combination with fluconazole (FLZ) on FLZ-resistant Candida albicans.
METHODThe minimum inhibitory concentrations (MIC) and sessile MIC80 (SMIC80) of EAHD and FLZ to FLZ-resistant C. albicans were determined by CLSI M27-A3 microdilution method, and the synergy of EAHD combined with FLZ were examined by the checkerboard microdilution assay. Agar plate-method was adopted to observe the rate of antifungal activity according to time-kill curve. HPLC and qRT-PCR were utilized to evaluate the changes of ergosterol content and expressions of related genes, respectively.
RESULTMICs of EAHD ranged from 156 to 1,250 mg · L(-1), those of FLZ from 256 to above 2,048 mg · L(-1) with FICI approximate 0.066 in combination; SMIC80 of EAHD were higher than 1,250 mg · L(-1), SMIC80 of FLZ were higher than 512 mg · L(-1) and up to above 2,048 mg · L(-1). Combination group also showed synergy effect except one group showing addition effect. The results of T-K experiment also confirmed obviously fungicidal effect when treated for 12 h. When compared with control groups, the ergosterol was reduced 85% and 50% in the treatments of combination and EAHD alone by HPLC, respective- ly. The expressions of ERG1, ERG2, ERG6, ERG7 and ERG11 were upregulated, and ACS1, ACS2, MET6 were downregulated when exposed to FLZ. The expressions of the above genes were downregulated by treatment of EAHD. The expressions of ERG2, ERG6, ERG11 were upregulated, while ERG1, ERG7, ACS1, ACS2, MET6 were downregulated in combination group.
CONCLUSIONThe combination of EAHD and FLZ exhibited synergy against FLZ-resistant C. albicans through decreasing the synthesis of ergosterol, and resulting in the breakage of cell membrane.
Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; growth & development ; metabolism ; Drug Resistance, Fungal ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Ergosterol ; biosynthesis ; Fluconazole ; pharmacology ; Microbial Sensitivity Tests
4.Effect of andrographolide on Candida albicans biofilm dispersion.
Gao-Xiang SHI ; Yuan-Yuan YAN ; Jing SHAO ; Ke-Qiao LU ; Meng-Xiang ZHANG ; Tian-Ming WANG ; Chang-Zhong WANG
China Journal of Chinese Materia Medica 2014;39(17):3339-3343
Along with the increase in fungal infections, Candida albicans prevention and control become the focus of anti-fungal infection at present. This study aims to discuss the effect monomer andrographolide (AG) on C. albicans biofilm dispersion. In the experiment, micro-well plates and medical catheter pieces were used to establish the C. albicans biofilm model. It was discovered by XTT assay and flat band method that 1 000, 500, 250 mg x L(-1) AG could impact the activity of C. albicans biofilm dispersion cells. The morphological structures of residual biofilms on catheter pieces were observed with scanning electron microscopy, which showed that 1 000, 500, 250 mg x L(-1) AG could induce C. albicans biofilm dispersion in a dose-dependent manner, and the dispersed cells were dominated by the yeast phase. According to the real-time fluorescence quantification PCR (qRT-PCR) test, AG could up-regulate HSP90 expression and down-regulate UME6 and PES1 expressions. This study demonstrates that AG could induce C. albicans biofilm dispersion to some extent.
Anti-Inflammatory Agents
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pharmacology
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Biofilms
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drug effects
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Candida albicans
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genetics
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physiology
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ultrastructure
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Diterpenes
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pharmacology
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Dose-Response Relationship, Drug
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Fungal Proteins
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genetics
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Gene Expression Regulation, Fungal
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drug effects
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HSP90 Heat-Shock Proteins
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genetics
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Microscopy, Electron, Scanning
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Reverse Transcriptase Polymerase Chain Reaction
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Time Factors
5.GNE gene mutation analysis in 5 patients with distal myopathy with rimmed vacuoles.
Xiang-hui LU ; Chuan-qiang PU ; Qiang SHI ; Wen-jing LUO ; Ke LI
Journal of Southern Medical University 2011;31(8):1421-1424
OBJECTIVETo investigate GNE gene mutations in 5 Chinese patients with distal myopathy with rimmed vacuoles (DMRV).
METHODSFive patients with typical clinical and pathological features of DMRV were studied. All the 11 coding exons and the flanking intron sequences of GNE gene were amplified by PCR and sequenced. Four family members of case 5 were also examined for GNE gene mutations.
RESULTSAll the patients were identified to have different GNE gene mutations: Cases 1-4 had complex heterozygous mutations and case 5 had homozygous mutation. Six reported mutations had been identified, including 1 nonsense mutation (p.R8X) and 5 missense mutations (p.D176V, p.I298T, p.A591T, P.A631V, and p.V696M). A novel mutation (c.317T>C, p.I106T) was identified in case 2.
CONCLUSIONThis is the first report of p.R8X, p.I298T, p.A591T and p.V696M mutations in GNE gene in Chinese population, and a novel mutation p.I106T was identified. These findings further expand the clinical and genetic spectrum of DMRV in China.
Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; DNA Mutational Analysis ; Distal Myopathies ; enzymology ; genetics ; Female ; Humans ; Male ; Molecular Sequence Data ; Multienzyme Complexes ; genetics ; Mutation ; genetics ; Mutation, Missense ; genetics ; Young Adult
6.Brief report of series study on Rongshui miniature pig as laboratory animal
Hehe SHI ; Gan CHEN ; Ke LIU ; Yunzhong LIU ; Xumin WANG ; Xiang GAO ; Zhengming HE ; Xiyong YU ; Liangxue LAI ; Xiaojiang TANG
Chinese Journal of Comparative Medicine 2015;(3):86-89
The provenances of Rongshui miniature pigs ( RMPs) were purchased from Rongshui, Guangxi, in 2012.130 RMPs were transported to Sanshui, Guangdong,China, which were breed according to the laboratory animal standards.83 RMPs were selected randomly from the first filial generations ( F1 ).The basic data were collected including breed characteristics, reproductive performance, grow curve, hematology, biochemical markers of serum and urine, organ coefficient, Chromosome analysis.According to the national and local standards, the quality control standards of RMP were set up including microbiological, parasitic, environment and facilities, fodder, pathology, genetic.The results showed that RMPs adapt to the climate of Guangdong.The natural mating and conceive rate was 88.3% with the pregnancy of 112 days.The average number of firstborn and still-born was 6.1 and 7.9 respectively.RMP was small body size with the adult body weight of 17.21 ±5.20 kg and 16.35 ±5.23 kg in female and male respectively.RMP was very tame.The mitochondrial genome analysis suggested RMP belonged a typical miniature pig breed in China, which is ancient than Lanyu pig.RMP could be breed as a new kind of laboratory animal.
7.Post-disaster medical rescue strategy in tropical regions
Xiang-Hui LI ; Shi-Ke HOU ; Jing-Chen ZHENG ; Hao-Jun FAN ; Jian-Qi SONG
World Journal of Emergency Medicine 2012;3(1):23-28
BACKGROUND: Earthquakes, floods, droughts, storms, mudslides, landslides, and forest wild fires are serious threats to human lives and properties. The present study aimed to study the environmental characteristics and pathogenic traits, recapitulate experiences, and augment applications of medical reliefs in tropical regions. METHODS: Analysis was made on work and projects of emergency medical rescue, based on information and data collected from 3 emergency medical rescue missions of China International Search and Rescue Team to overseas earthquakes and tsunamis aftermaths in tropical disaster regions Indonesia-Aceh, Indonesia-Yogyakarta, and Haiti-Port au Prince. RESULTS: Shock, infection and heat stroke were frequently encountered in addition to outbreaks of infectious diseases, skin diseases, and diarrhea during post-disaster emergency medical rescue in tropical regions. CONCLUSIONS: High temperature, high humidity, and proliferation of microorganisms and parasites are the characteristics of tropical climate that impose strict requirements on the preparation of rescue work including selective team members suitable for a particular rescue mission and the provisioning of medical equipment and life support materials. The overseas rescue mission itself needs a scientific, efficient, simple workflow for providing efficient emergency medical assistance. Since shock and infection are major tasks in post-disaster treatment of severely injured victims in tropical regions, the prevention and diagnosis of hyperthermia, insect-borne infectious diseases, tropic skin diseases, infectious diarrhea, and pest harms of disaster victims and rescue team staff should be emphasized during the rescue operations.
8.Effect of andrographolide on quorum sensing and relevant virulence genes of Candida albicans.
Yuan-yuan YAN ; Gao-xiang SHI ; Jing SHAO ; Ke-qiao LU ; Meng-xiang ZHANG ; Tian-ming WANG ; Bin WANG ; Chang-zhong WANG
China Journal of Chinese Materia Medica 2015;40(2):292-297
OBJECTIVETo investigate the effect of andrographolide (AG) on quroum sensing (QS) and relevant virulence genes of Candida albicans.
METHODGas-chromatography-mass spectrometry (GC-MS) was applied to detect the changes in the content of farnesol and tyrosol in C. albicans intervened by AG. The real-time quantitative PCR (qRT-PCR) was adopted to inspect the expressions of relevant virulence genes such as CHK1, PBS2 and HOG1 regulated by QS.
RESULTAt 2 h after the growth of C. albican, the farnesol and tyrosol secretions reduced, without notable change after intervention with AG. The secretions were highest at 12 h and decreased at 24 h. After the intervention with different concentrations of AG, the farnesol content reduces, whereas tyrosol increased, indicating a dose-dependence, particularly with 1 000 mg x L(-1) AG. qRT-PCR revealed that 1 000 mg x L(-1) AG could down-regulate CHK1 by 2.375, 3.330 and 4.043 times and PBS2 by 2.010, 4.210 and 4.760 times, with no significant change in HOG1.
CONCLUSIONAG could inhibit the farnesol secretion, promote the tyrosol secretion and down-regulate QS-related virulence genes CHK1 and PBS2 expressions.
Candida albicans ; drug effects ; genetics ; physiology ; Diterpenes ; pharmacology ; Farnesol ; analysis ; metabolism ; Gas Chromatography-Mass Spectrometry ; Genes, Fungal ; Phenylethyl Alcohol ; analogs & derivatives ; analysis ; metabolism ; Quorum Sensing ; drug effects ; Real-Time Polymerase Chain Reaction ; Virulence ; genetics
9.Neuroendoscope-assisted micro-invasive surgical treatment of hypertensive thalamic hemorrhage
Yi-Zhao CHEN ; Ru-Xiang XU ; Yong-Geng NIE ; Li-Ke SAI ; Xiang-Yu WANG ; Cheng-Yi LUO ; Shi-Zhong ZHANG ; Yi-Quan KE
Chinese Journal of Neuromedicine 2010;09(9):936-939
Objective To develop a minimally invasive operating technique for the treatment of hypertensive thalamic hemorrhage. Methods The clinical data of 15 patients with hypertensive thalamic hemorrhage performed neuroendoscope-assisted micro-invasive surgical treatment in our hospitals from July 2007 to June 2010 were retrospectively analyzed; their treatment efficacy were also concluded. Results The mean operation time of these patients was (1.5±0.4) h and the amount of blood loss was 30-40 mL; the mean clearance rate of hematoma in the thalamus was (86.2 ±7.9)percentage. Patients were followed up and evaluated by Glasgow outcome scale for at least 3 months.Three patients (21.4%) showed good recovery, 4 (28.6%) moderate disability, 4(28.6%) severe disability and 2 (14.3%) vegetative survival; 1 patient (7.1%) died. Conclusion Neuroendoscope-assisted micro-invasive surgical treatment is a fast and minimally invasive operating technique with little blood loss in the treatment of hypertensive thalamic hemorrhage.
10.Three-dimensional reconstruction of CT imaging in endoscopic surgery of patients with hypertensive intracerebral hemorrhage
Yi-Zhao CHEN ; Bo-Miao LIN ; Ru-Xiang XU ; Yong-Geng NIE ; Li-Ke SAI ; Xiang-Yu WANG ; Shi-Zhong ZHANG ; Yi-Quan KE
Chinese Journal of Neuromedicine 2011;10(3):280-283
Objective To develop a simple, fast and accurate preoperative planning method for endoscopic surgery of patients with hypertensive intracerebral hemorrhage (HICH).Methods Eighteen patients with HICH, admitted to our hospital from June 2008 to August 2010, were performed endoscopic minimally invasive surgery; CT three-dimensional reconstruction was employed to locate the intracerebral hematoma and select the appropriate endoscopic approach before the endoscopic surgery.The clinical data and treatmem efficacy were analyzed.Results According to the results of CT three-dimensional reconstruction, our neurosurgeons could design the best endoscopic approach; the three-dimensional relationship between intracerebral hematoma and scalp markers was shown directly and accurate positioning of the location of drilling was achieved; therefore, the time for preoperative preparation, anesthesia and operation was shortened. The mean operating time of these 18 patients was about 1.5 h; the volume of blood loss was only 30-40 mL; and the evacuation ratio was about 89.2%.After the elimination of hematoma, the brain tissues were flabby, so decompressive craniectomy was not needed. Conclusion CT three-dimensional reconstruction is a simple, fast and accurate preoperative planning method for endoscopic surgery of patients with HICH.