1.Application of in vitro bionic digestion and biomembrane extraction for metal speciation analysis, bioavailability and risk assessment in lianhua qingwen capsule.
Lu-Xiu LIN ; Shun-Xing LI ; Feng-Ying ZHENG
China Journal of Chinese Materia Medica 2014;39(12):2330-2335
One of the causes of the high cost of pharmaceuticals and the major obstacles to rapidly assessing the bioavailability and risk of a chemical is the lack of experimental model systems. A new pre-treatment technology, in vitro bionic digestion was designed for metal analysis in Lianhua Qingwen capsule. The capsule was digested on 37 degrees C under the acidity of the stomach or intestine, and with the inorganic and organic compounds (including digestive enzymes) found in the stomach or intestine, and then the chyme was obtained. Being similar to the biomembrane between the gastrointestinal tract and blood vessels, monolayer liposome was used as biomembrane model Affinity-monolayer liposome metals (AMLMs) and water-soluble metals were used for metal speciation analysis in the capsule. Based on the concentration of AMLMs, the main absorption site of trace metals was proposed. The metal total contents or the concentration of AMLMs in the capsule were compared to the nutritional requirements, daily permissible dose and heavy metal total contents from the "import and export of medicinal plants and preparation of green industry state standards". The metal concentrations in the capsule were within the safety baseline levels for human consumption. After in vitro bionic digestion, most of trace metals were absorbed mainly in intestine. The concentration of As, Cd, Pb was 0.38, 0.07, 1.60 mg x kg(-1), respectively, far less than the permissible dose from the "import and export of medicinal plants and preparation of green industry state standards".
Biological Availability
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Capsules
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adverse effects
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pharmacokinetics
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Digestion
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Drugs, Chinese Herbal
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adverse effects
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pharmacokinetics
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Humans
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Metals, Heavy
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adverse effects
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pharmacokinetics
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Models, Biological
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Stomach
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metabolism
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Trace Elements
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adverse effects
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pharmacokinetics
2.Fas Ligand gene transfection induces rat mast cells apoptosis
Qingyu XIU ; Danrong YANG ; Huanxing HAN ; Zhaoquan SHI ; Huiqi LU ; Lin ZHOU
Chinese Journal of Immunology 1985;0(03):-
Objective:To show whether the Fas Ligand gene induces mast cells apoptosis.Methods:RT-PCR was used to amplify the gene of rat Fas ligand extracelluar domain and transmembrane domain and cloned it into eukaryotic expression plasmid pcDNA3.1.Transcent transfect RBL-2H3,the expression of Fas ligand RBL-2H3 was detected by RT-PCR、Western blot.The Annexin V FCM was used to detect the RBL-2H3 apoptosis after the transfection of Fas Ligand.Results:It is successful to obtain the gene of rat Fas Ligand extracellular domain and transmembrane segment,cloning it into pcDNA3.1,FasL was expressed on the surface of RBL-2H3 and it's supernatant after the transfection of pcDNA3.1/FasL.The cell start to be apoptosis.Conclusion:Our study reveals that Fas Ligand gene transfection in RBL-2H3 can effectively induced apoptosis.It is a promising strategy for Fas Ligand to be used in the therapy of allergic disease. [
3.Bispectral index in predicting the prognosis of patients with coma in intensive care unit
Lin DOU ; Hong-Mei GAO ; Ling LU ; Wen-Xiu CHANG
World Journal of Emergency Medicine 2014;5(1):53-56
BACKGROUND:The bispectral (BIS) index is a processed electroencephalogram (EEG) parameter with extensive validation and demonstrated clinical utility. The study aimed to investigate the correlation between the BIS index and the prognosis of patients with coma in the ICU. METHODS:A total of 208 patients with coma in the ICU were enrolled in this study. According to the BIS value, the patients were divided into four groups:group I, BIS value 0 to 20; group Ⅱ, BIS value 21 to 40; group Ⅲ, BIS value 41 to 60; and group IV, BIS value greater than 60. The difference in BIS values with the differences in prognosis of patients with coma was compared between the four groups, and the prognosis of patients with coma was stratified into consciousness, coma, vegetative state, and brain death. Subsequently, the best cut-off score of BIS values calculated for determining the correlation between BIS value and mental state was proposed. RESULTS:There are no significant differences in the age and APACHE Ⅱ scores between the four groups (P>0.05). An inverse correlation was observed between BIS value and mental state (r=–0.749, P=0.00). According to the ROC curve, as BIS value was greater than 42.5, there were higher sensitivity and specificity in conscious-coma patients. CONCLUSION:BIS value is correlated with the prognosis of patients with coma in ICU, and BIS value can be a useful marker for estimating the prognosis of comatose patients.
4.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.Stem cell characteristics and islet differentiation potential of human amniotic epithelial cells.
Jian WANG ; Lin PENG ; Guang-xiu LU
Journal of Southern Medical University 2011;31(9):1484-1487
OBJECTIVETo establish a method for isolating and culturing human amniotic epithelial cells (hAECs) in a serum-free medium and investigate their transdifferentiation ability into islet-like cells.
METHODSThe culture condition of hAECs was optimized using DMEM with different supplements. The genetic stability of the tenth-passage cells was assessed by chromosome analysis and G-banding method. The stem cell characteristics of the cells were identified by examination of the surface markers using immunofluorescence methods. The endocrine-related genes and hormones of the cells were tested after induced differentiation into islet-like cells.
RESULTSThe hAECs allow stable passaging in the presence of 10 ng epidermal growth factor (EGF) in the culture medium. After 10 passages, the cells maintained a normal karyotype and G-banding profile. The hAECs expressed many multi-potent stem cell markers, including SSEA4, TRA-1-60, and TRA-1-81. After induced differentiation, the endocrine-related genes were expressed in the islet-like cells, including PDX1, ngn3, insulin and glucagon. Insulin secretion increased in the differentiated islet-like cells in response to high glucose exposure.
CONCLUSIONWe established a method for isolating and expanding the hAECs in a serum-free medium. hAECs possess stem cell characteristics and can be induced to differentiate into islet-like cells in vitro.
Amnion ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cells, Cultured ; Epithelial Cells ; cytology ; secretion ; Humans ; Insulin ; secretion ; Islets of Langerhans ; cytology ; Stem Cells ; cytology ; secretion
6.Stress analysis of the all-ceramic crown with a numerical approach.
Cheng-Lin LU ; Yan-Ling WU ; Xiu-Yin ZHANG ; Dong-Sheng ZHANG
Chinese Journal of Stomatology 2009;44(3):155-158
OBJECTIVETo analyze the stress distribution in all-ceramic crowns when it was subjected to load.
METHODSA 3D numerical model of the all-ceramic crown of the right mandibular first molar was generated from scanned CT images. Finite element analysis (FEA) was used to evaluate the stress distribution in the all-ceramic crown when it was subjected to 5 load conditions.
RESULTSStress distributions under 5 loading conditions were obtained. Stress concentrations were generally found near the loading areas at the veneer and at the lower surface of the core beneath the loading areas. In the 5 loading conditions, it was found that when the crown was loaded with vertical concentrated load, the tensile stresses around the shoulder areas were uniform, while stress concentration with maximum of 32.25 MPa was found at the shoulder areas in lingual-buccal direction when loads were applied at an angle of 10 degrees with the tooth axis on the buccal side. The masticatory load which was applied at 20 degrees with the tooth axis on the buccal side would cause stress concentration at the shoulder in mesial-distal direction. The maximum value could reach 11.29 MPa.
CONCLUSIONSThe occlusal surface of the all-ceramic crown must be trimmed to increase multiple contact zones with the opposite surfaces in antagonist teeth to avoid excessive concentrated stress that may cause crown failure.
Crowns ; Dental Porcelain ; Dental Stress Analysis ; methods ; Finite Element Analysis ; Materials Testing
7.Cloning and expression analysis of pathogenesis-related protein 1 gene of Panax notoginseng.
Rui-Bo LI ; Xiu-Ming CUI ; Yu-Zhong LIU ; Zhi-Gang WU ; Shu-Fang LIN ; Ye SHEN ; Lu-Qi HUANG
Acta Pharmaceutica Sinica 2014;49(1):124-130
By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence
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Cloning, Molecular
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Escherichia coli
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metabolism
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Molecular Weight
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Open Reading Frames
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genetics
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Panax notoginseng
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chemistry
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Phylogeny
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Plant Proteins
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genetics
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metabolism
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Plants, Medicinal
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chemistry
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Reverse Transcriptase Polymerase Chain Reaction
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Sequence Alignment
8.A study on the influenza surveillance program in Tianjin, 2004.
Jie-xiu WANG ; Ai-lan SHAN ; Lu GAO ; Lin LI
Chinese Journal of Epidemiology 2005;26(11):848-850
OBJECTIVETo explore the trend of influenza epidemics, predominate strains of the virus in Tianjin city and to analyze the economic impact of vaccine-based interventions for the prevention and control of influenza.
METHODSData on epidemiological studies and on virus surveillance was gathered. Monte Carlo mathematical simulation modelwas used for data analyses.
RESULTSFrom 29, December, 2003 to 2, January, 2005, the proportion of influenza-like infection cases was accounted for 8.93% of the total number of patients from the outpatient departments of 4 general hospitals in Tianjin. The proportion reached its peak from November to February and it was accounted for 9.39% in epidemic period, which was significantly different from that in the non-epidemic period (u = 15.53, P< 0.05). There was 56 strains of influenza virus isolated in which 45 were indentified as A(H3N2) and 11 as B with a total positive rate as 13.21%. Predominant strain was found bing type A(H3N2). The positive rate was 15.41% in the epidemic period, which was significantly different from that in the non-epidemic period (u = 2.519, P < 0.05). The cost per visit ranged from 475.93 to 581.69 Yuan (RMB) with an average cost of 528.81 Yuan. When the attack rate increased to 30 percent, the positive mean net returns would have been 24 million Yuan among the 0-19 age group.
CONCLUSIONInfluenza did not seem to be prevalent in Tianjin from 29, December, 2003 to 2, January, 2005. The main type of influenza was type A(H3N2). It is necessary to pay attention to the all-year round surveillance program due to the transformation of influenza type A and the slight increase of incidence in summer.
Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Health Care Costs ; statistics & numerical data ; Humans ; Infant ; Infant, Newborn ; Influenza A Virus, H3N2 Subtype ; physiology ; Influenza B virus ; physiology ; Influenza, Human ; economics ; epidemiology ; immunology ; virology ; Middle Aged ; Population Surveillance ; Seasons ; Viral Vaccines ; immunology
9.Behavioral alterations and demyelization of the corpus callosum in the mouse model of MK-801 induced schizophrenia
Yun XIU ; Lei ZHANG ; Xuan QIU ; Lin CHEN ; Wei LU ; Chao PENG ; Guohua CHENG ; Fenglei CHAO ; Yong TANG
Chinese Journal of Nervous and Mental Diseases 2013;(11):641-645
Objective To explore the role of white matter injuries in the schizophrenia induced by the NMDA re-ceptor antagonist. Methods Adult male C57BL/6J mice (8 week old) were equally divided into four groups. One group was sub-chronically treated with saline solution, and the other three groups were intraperitoneally treated with MK-801 at dose of 0.025 mg/mL (M1), 0.050 mg/mL (M2) and 0.100 mg/mL (M3) in a volume 10 ml per kilogram body weight. All ani-mals were tested using Morris water maze at the 9th-15th day and using the Hole Board exploration as well as Rota Rod performance tests on the 16th day. The myelin basic protein (MBP) and the ultrastructure of the myelin sheaths in the cor-pus callosum were then examined using immunohistochemical methods, transmission electron microscope technique and stereological methods. Results The repeated sub-chronic MK-801 treatment did not induce impairment of spatial learning and memory in Morris water maze. The MK-801 treatment at 0.25 mg/kg and 1.00 mg/kg but not at 0.50 mg/kg resulted in less exploration to a new environment. The myelin staining with anti-MBP antibody was less intense in all three schizo-phrenic groups when compared to saline control group (P<0.01). Furthermore, MK-801 treatment caused pathological al-terations of the myelin sheaths including segmental demyelination of myelinated fibers and splitting of myelin sheath lamel- lae in schizophrenic groups. The ratio of the injured myelinated nerve fibers in the corpus callosum of MK-801 treated mice [M3 group, (22.42 ± 4.24)%] was significantly higher when compared to the control mice [(3.84 ± 1.35)%,P<0.01)]. Conclusions The present study demonstrated the white matter damages, mainly low MBP expression and segmental demye-lization in the corpus callosum in the mice sub-chronic treated with MK-801, indicating that the white matter changes might be involved in the schizophrenia induced by NMDA antagonist.
10.Inhibition effect of Heme oxygenase-1 activator Copp on irradiation-induced endothelial cell apoptosis
jing, YU ; guo-hua, WU ; qing, LIN ; ren-hua, ZHOU ; dong-qing, LU ; jia, WANG ; qian, LIU ; xiu-yan, FEN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(12):-
Objective To explore protective effect of Heme oxygenase-1(HO-1) on irradiation-induced endothelial cell apoptosis.Methods Human endothelial cell line EA.hy926 were administered with or without HO-1 activator Copp and/or HO-1 inhibitor Znpp,respectively.Then,cells were treated with or without 8 Gy radiation.The HO-1 protein expression of cells were assessed with Western blotting and apoptosis of cells treated with irradiation were evaluated with flow cytometry.Moreover,cytochrome C releasing into cytosol were also determined by Western blotting. Results In PBS+R group,HO-1 protein expression of EA.hy926 was low posterior to irradiation.When cells were preconditioned with Copp and/or Znpp,then recieved with 8Gy irradiation,the HO-1 protein expression of EA.hy926 increased significantly in comparision with the PBS+R group(P