2.Isolation and 16S rRNA Gene Sequence Analysis of a Thermotolerant Halophilic Bacterium
Ying XIE ; Lian-Bing LIN ; Xiu-Ling JI ; Yun-Lin WEI ;
Microbiology 1992;0(02):-
A strain of thermotolerant halophilic bacteria YJ0238 was isolated from the salt well at 47℃ in the Kangning countryside where was located at the side of Lancang River in the Tibet Autonomous Region. Some physiological and biochemical properties were characterized.16S rRNA gene sequence of YJ0238 was amplified by PCR,and its nucleotide sequence was determined. Based on it’s physiological and biochemical properties,homology and phylogenetic analysis of 16S rRNA gene sequence,strain YJ0238 was identified as a subspecies of the species Idiomarina zobellii. The GenBank accession number of the 16S rRNA gene sequence of strain YJ0238 is EF693953. Until now,there were few reports on the study of high-temperature and high-salt microbial in domestics. The results of this study will provide research material and information for further studies in this area.
3.Identification of a Thermoacidophilic Sulfolobus sp. Isolated from a Hot Spring in Tengchong Rehai
Bo CHEN ; Yun-Lin WEI ; Shen-Rong JING ; Xiu-Ling JI ; Yue-Qing LU ; Lian-Bing LIN ;
Microbiology 2008;0(12):-
An extremely thermoacidophilic isolate K4-1 was obtained from an acidic hot spring in Teng- chong Rehai, Yunnan province. Morphology, growth characteristics, utilization of carbon compounds, en- ergy sources and 16S rRNA gene sequence of K4-1 were studied. Cells of K4-1 are irregular cocci with monotrichous flagella. The strain grew aerobically in either a lithotrophic or a heterotrophic mode. Growth on elemental sulfur occurred through oxidation of sulfur. It grew optimally at 75?C and pH 3.5. On the basis of 16S rRNA gene sequence similarity, strain K4-1 was shown to belong to genus Sulfolobus, being related to the type strains of genus Sulfolobus (86.6%~94.3% similarity), and being most closely related to strain Sulfolobus tengchongensis RT8-4 (98.9% similarity). The GenBank accession number of strain K4-1 16S rRNA gene sequence is EU729124.
5.Construction of the Bac-to-Bac System of Bombyx mori Nucleopolyhedroviru
Jin-shan, HUANG ; Bi-fang, HAO ; Xiu-lian, SUN ; Fei, DENG ; Hua-lin, WANG ; Zhi-hong, HU
Virologica Sinica 2007;22(3):218-225
To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus (BmNPV), a transfer vector was constructed which contained an Escherichia coli (E. coli) mini-F replicon and a lacZ: attTN7: lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene. B. mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate recombinant viruses by homologous recombination in vivo. The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E. coli DH10B. Recombinant bacmids were screened by kanamycin resistance, PCR and restriction enzyme (REN) digestion. One of the bacmid colonies, BmBacJS13, which had similar REN profiles to that of wild-type BmNPV, was selected for further research. To investigate the infectivity of BmBacJS13, the polyhedrin gene was introduced into the bacmid and the resultant recombinant (BmBacJS13-ph) was transfected to BmN cells. The budded viruses were collected from the supernatant of the transfected cells and used for infecting BmN cells. Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV. Bio-assays indicated that BmBacJS13-ph was also infectious to B. mori larvae.
6.Using westgard's method evaluation decision chart for judging method performance of routine biochemistry items on Roche Modular PPI testing system
Xiu-Ming ZHANG ; Song-Bai ZHENG ; Lei SUN ; Pei-Feng KE ; Ji MA ; Wen ZHANG ; Lian-Ying LIN ;
Chinese Journal of Laboratory Medicine 2001;0(01):-
Objective To judge method performance of routine biochemistry parameters on Roche Modular PPI testing system by using westgard's method evaluation decision chart.Methods We assessed imprecision(CV)from internal quality control and inaccuracy(bias)from external quality control evaluation.Combined estimates of imprecision and inaccuracy by plotting imprecision as the x-coordinate and inaccuracy as the y-coordinate to locate an expected operating point of every item on the chart.By comparing this operating point with allowable total errors(TEa),we can decide whether the performance is acceptable or not.Results In the 27 different parameters tested,imprecision and bias of calcium were 0.08 mmol/L and 0.06 mmol/L respectively,its performance was marginal.The imprecision of creatinine,urea,glucose, sodium,chloride and phosphorus were 3.20%,2.13%,1.52%,0.89 mmol/L,1.10% and 1.55%,the bias were 4.79%,0.96%,4.63%,0.80 mmol/L,1.74% and 4.13% respectively,their performance was good.M1 other 20 items were of excellence performance.Conclusions Routine biochemistry parameters on Roche Modular PPI testing system possessed good precision and accuracy,and their performance were acceptable.To judge method performance of biochemistry testing system by using westgard' s method evaluation decision chart was easy to do and suited for clinical laboratory.
7.Clinical application analysis of unstable angina medications
Xiu-Qin ZHANG ; Hong-Lin HUANG ; Jin-Lian BI
The Chinese Journal of Clinical Pharmacology 2015;(21):2158-2160
Objective To analysis the clinical applications of unstable an-gina medications.Methods Retrospective analysis of the original re-cords of 327 patients with unstable angina pectoris and evaluation of the use and rationality of the medications using the indicators of defined daily dose(DDD), frequency of drug use(DDDs) and drug utilization index ( DUI) .Results Top three of the most frequently used anti-ischemic drugs were metoprolol tartaric acid ( 40.06%) , glyceryl trinitrate ( 33.03%) and metoprolol succinate sustained -release tablets ( 25.99%) .In addition, 97.54% of patients received anti -platelet therapy, 12.23%received anti-coagulation therapy, and 95.41% re-ceived treatment of statins.As for DUI, except of aspirin and clopi-dogrel, other drugs were ≤1.Conclusion Except that the intensity of anti-coagulation therapy is not enough, the use of other drugs was rea-sonable and consistent with the guidelines basically.
8.Changes of Cytokines Measurement in Serum and Cerebrospinal Fluid in Asphyxia Ne onates and Its Relationship with Prognostic
fa-lin, XU ; chang-lian, ZHU ; xiu-yong, CHENG ; li-ting, JIA ; lin-lin, GENG ; jun-ying, QIAO ; sheng-hai, HU
Journal of Applied Clinical Pediatrics 2004;0(08):-
Objective To study the dynamic changes of cytokines including tumor necrosis factor-alpha (TNF-?),interleukin-6 (IL-6),IL-8 in serum and cerebrospinal fluid (CSF) of asphyxia neonates,and to analyze the relationship between cytokines levels and severity of brain damage and neurological outcome. Methods The concentrations of TNF-?,IL-6,IL-8 in serum and CSF were measured by radioimmunoassay in 63 asphyxia neonates. Neurological development was evaluated at 12 months by children′s developmental scale of china.Results The serum concentrations of TNF-?, IL-6,IL-8 were significantly higher in asphyxiated neonates than those in the controls,and they were correlated with the degree of encephalopathy. The level of serum TNF-? was hig-(hest) at the first day and IL-6 was highest at the third day. There was no marked dynamic changes within 5 days in serum IL-8 level. The concentrations of TNF-?,IL-8 in CSF were higher at the first and the third day.The dynamic changes of IL-6 in CSF were similar in serum and they were positively correlated. The serum concentration of IL-6 in severe brain injury group was much higher than those of normal and mild group.The CSF concentration of IL-6 in severe brain injury group was much higher than that of normal group. The CSF concentration of IL-8 in severe brain injury group was much higher than those of the normal and mild group. Conclusions The concentrations of TNF-?,IL-6 and IL-8 are increased both in serum and CSF in asphyxiated neonates which are correlated with severity of hypoxic-ischemic encephalopathy. Cytokine-mediated inflammatory reactions may participate in the mechanism of hypoxic-ischemic brain injury after asphyxiaion.The concentration of IL-6 in serum and IL-6, IL-8 in CSF are correlated with the neurological outcome.
9.Construction of promoter probe vector for a cold-adapted bacterium, Acinetobacter sp. DWC6.
Yun-Lin WEI ; Lian-Bing LIN ; Xiu-Ling JI ; Shen-Rong JING
Chinese Journal of Biotechnology 2007;23(3):530-534
In order to build a protein expression system in a cold-adapted bacterium Acinetobacter sp. DWC6, a promoter probe vector was constructed based on the plasmid pBR322. A fragment containing the promoter of the beta-lactamase gene (the ampicillin resistance gene) in pBR322 was eliminated and replaced by a fragment comprizing a kanamycin resistance gene amplified from pJRD215. DNA fragment harboring in the Acinetobacter species specific ori was also inserted into the plasmid pBR322 to construct a promoter probe vector named pBAP1, which could replicate both in E. coli and in Acinetobacter sp. DWC6. The promoter selection library was constructed by randomly inserting genomic DNA fragment of Acinetobacter sp. DWC6 at upstream of reported gene, and target promoters were screened from genomic library on ampicillin selection plates. The function of pBAP1 and isolated promoters were determined by detection of the ampicillin sensitivity and the expression level of beta-lactamase in the host cell.
Acinetobacter
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genetics
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Adaptation, Physiological
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Bacterial Proteins
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genetics
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metabolism
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Cloning, Molecular
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Cold Temperature
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DNA, Bacterial
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genetics
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Electrophoresis, Polyacrylamide Gel
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Genetic Vectors
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genetics
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Models, Genetic
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Plasmids
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genetics
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Promoter Regions, Genetic
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genetics
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beta-Lactamases
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genetics
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metabolism
10.Study on content of two sesquiterpenes in Alpinia oxyphylla.
Xiu-xiang LV ; Lei HOU ; Wen-hua HUANG ; Bao-lin GUO ; Lian-xue ZHANG ; Jing-guang YU
China Journal of Chinese Materia Medica 2012;37(17):2589-2593
OBJECTIVETo develop an HPLC method for determining oxyphyllenodiol A (1) and teuhetenone A (2) contained in Alpinia oxyphylla and to compare the contents of the two components contained in medicinal materials and prepared herbal medicines in pieces sold in the market and different fractions.
METHODHPLC and Waters sunfire C18 column (4. 6 mm x 250 mm, 5 microm) were adopted for gradient elute with the mobile phase of acetonitrile and water. The flow rate was 1.0 mL x min(-1) and the detection wavelength was 250 nm.
RESULT1 and 2 showed a good linear relationship within the range of 0.1296 - 0.8640 microg and 0.1635 - 1.0900 microg respectively, with the average recoveries of 99.08% and 97.80%. Their content ranges were 0.0059% - 0.0149% and 0.0080% - 0.0164% in different samples. The mean value of 1 and 2 were 0.0085% and 0.0104% in the whole fruits, and 0.0137% and 0.0157% in the seeds. They were undetected in the nutshells.
CONCLUSIONThe method is so precise, accurate and highly reproducible that it can be used to determine the contents of oxyphyllenodiol A and teuhetenone A in A. oxyphylla. The contents of the two components are mainly extracted from the seeds, with certain difference among different samples. There are a higher contents and no significant difference in the salted and raw seeds.
Alpinia ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Sesquiterpenes ; analysis ; isolation & purification