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Objective:To construct core medical skill index system for helicopter medical rescue personnel by Delphi expert consensus method.Methods:This study combined literature review and two rounds of Delphi expert correspondence to construct the core skill index system. Total of 22 experts from relevant disciplines were selected purposively to conduct Delphi expert consultation.Results:The effective questionnaire recovery rates of the two rounds of Delphi expert correspondence was 100%; the authority coefficient of experts was 0.87; the Kendall coordination coefficients of the importance of the first and second level indexes were 0.380 and 0.338 in the first round of correspondence, and then 0.415 and 0.357 in the second round. the Kendall coordination coefficients of the feasibility of the second level indexes were 0.347 in the first round of correspondence and 0.354 in the second round. The final core skill index system for helicopter medical rescue personnel includes 4 primary indicators: core medical skill for diseases or trauma in respiratory system, circulatory system, sports system and other system, and 32 secondary indicators.Conclusions:The core medical skill index system for helicopter medical rescue personnel constructed through two rounds of Delphi expert correspondence is scientific and reasonable, which can provide a scientific basis to accurately understand, describe, analyze, and evaluate the competence level of helicopter medical rescue personnel, as well as serve as a content framework for training programs.
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Objective:To observe and analyze the gene mutation and clinical phenotype of patients with cone and rod dystrophy (CORD).Methods:A pedigree investigarion. Two CORD pedigrees including 2 patients and 6 family members were enrolled in Ningxia Eye Hospital of People' Hospital of Ningxia Hui Automous Region for this study. The patients were from 2 unrelated families, all of whom were probands. Take medical history with best-corrected visual acuity (BCVA), color vision, slit lamp microscopy, indirect ophthalmoscopy, fundus color photography, optical coherence tomography (OCT), autofluorescence (AF), fluorescein fundus angiography (FFA), electroretinogram (ERG). The peripheral venous blood of patients and their parents was collected, whole genome DNA was extracted, Trio whole genome exome sequencing was performed, Sanger verification and pedigree co-segregation were performed for suspected pathogenic mutation sites. According to the law of inheritance, family history was analyzed to establish its genetic type. Mutational loci pathogenicity was analyzed according to the American College of Medical Genetics (ACMG) guidelines and 4 online tools.Results:Two CORD families showed autosomal recessive inheritance. The proband of pedigree 1 was female, 49 years old. Binocular vision loss with photophobia lasted for 9 years and night blindness for 4 years. The BCVA of right eye and left eye were 0.03 and 0.06, respectively. The results of ERG showed that the amplitudes of dark adaptation 0.01 b-wave and dark adaptation 3.0 a-wave and b-wave in both eyes were slightly decreased, and the amplitudes of light adaptation 3.0 a-wave and b-wave were severely decreased. The proband of pedigree 2 was male, 30 years old. Vision loss in both eyes for 4 years. Denying a history of night blindness. The BCVA of right eye and left eye were 0.3 and 0.2, respectively. The results of ERG showed that the amplitudes of dark adaptation 0.01 b-wave and dark adaptation 3.0 a-wave and b-wave in both eyes were slightly decreased, and the amplitudes of light adaptation 3.0 a-wave and b-wave were severely decreased. The color of optic disc in both eyes was light red, the macular area was atrophic, the foveal reflection disappeared, and the peripheral retina was punctate pigmentation. The main fundus changes in 2 patients were macular atrophy. The proband of pedigree 1 carried compound heterozygous variations c.439-2A>G (M1) and c.676delT (p.F226fs) (M2) on CDHR1 gene. Her father and mother carried M2 and M1 heterozygous mutations, respectively. The proband of pedigree 2 carried compound heterozygous variations c.2665dupC (p.L889fs) (M3) and c.878T>C (p.L293P) (M4) on C2orf71 gene. His father and mother carried M4 and M3 heterozygous mutations, respectively. According to ACMG guidelines and on line tools, 4 variations were considered as pathogenic level. Conclusions:M1 and M2 of CDHR1 gene and M3 and M4 of C2orf71 gene are new pathogenic mutations of CORD. All patients presented with the clinical phenotype of decreased visual acuity and macular atrophy.
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Gene editing technology can be used to modify the genome of Escherichia coli for the investigation of gene functions, or to change the metabolic pathways for the efficient production of high-value products in engineered strains with genetic stability. A variety of gene editing technologies have been applied in prokaryotes, such as λ-Red homologous recombination and CRISPR/Cas9. As a traditional gene editing technique, λ-Red recombination is widely used. However, it has a few shortcomings, such as the limited integration efficiency by the integrated fragment size, the cumbersome gene editing process, and the FRT scar in the genome after recombination. CRISPR/Cas9 is widely used for genome editing at specific sites, which requires specific DNA segments according to the editing site. As the understanding of the two technologies deepens, a variety of composite gene editing techniques have been developed, such as the application of λ-Red homologous recombination in combination with homing endonucleaseⅠ-SceⅠ or CRISPR/Cas9. In this review, we summarized the basic principles of common gene editing techniques and composite gene editing techniques, as well as their applications in Escherichia coli, which can provide a basis for the selection of gene editing methods in prokaryotes.
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CRISPR-Cas Systems/genetics , Escherichia coli/genetics , Gene Editing , Homologous Recombination , TechnologyРеферат
Objective To investigate the effect of microRNA-1183 on proliferation and metastasis on gastric cancer cells and to explore the role of microRNA-1183 and CBL-B signaling pathways in this process. Methods MGC803 cells were transfected with a microRNA-1183 mimic. Real-time PCR detected the expression of microRNA-1183 in gastric cancer cell line MGC803. MTT detected the proliferative effect of microRNA-1183 on MGC803 gastric cancer cells. A Transwell assay detected the effect of microRNA-1183 on the metastasis of MGC803 gastric cancer cells. A dual luciferase reporter assay detected the binding ability between microRNA-1183 and CBL-B. The expression of the protein was tested by Western blotting. Results MTT assay results showed that microRNA-1183 promoted the proliferation of MGC803 cells. Transwell assay results revealed that microRNA-1183 promoted the metastasis of MGC803 cells. The results of BLAST contrast analysis show that CBL-B is one of the target genes of microRNA-1183. Western blotting analysis showed that the mimic microRNA-1183 inhibited the expression of CBL-B. A dual luciferase reporter assay showed that CBL-B was the target gene of microRNA-1183. A CBL-B knockdown promoted the proliferation and metastasis of MGC803 cells. microRNA-1183 promoted the proliferation and metastasis of MGC803 cells by inhibiting the expression of CBL-B. Conclusion microRNA-1183 can inhibit the proliferation and metastasis of gastric cancer cell lines by inhibiting the expression of CBL-B.
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Objective To investigate the effect of Exosomes derived from lung cancer cells on the migration of secretory cells and homologous tumor cells and to explore the role of PI3K/Akt and SRC signaling pathways in this process.Methods Exosomes were isolated from the supematant post density gradient centrifugation of A549,lung cancer cells.Morphology of the Exosomes was studied using transmission electron microscopy.Protein expression was analyzed using Western blotting.Cell migration was analyzed by a transwell assay.Results The double-membrane-bound Exosomes appeared as discal-shaped structures,30-100 nm in diameter.Western blotting showed that CD9 was abundant in the Exosomes.The Exosomes promoted the migration of A549 cells and their homologous tumor cells,HCC827 in a dose-dependent manner,accompanied by the activation of Akt and SRC.Conclusion The Exosomes derived from A549 (lung cancer) cells promote the migration of the secreting cells and the homologous tumor cells.The mechanism may be correlated with the activation of Akt and SRC.
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Objective To investigate the distribution of traditional Chinese medicine (TCM) syndrome types of functional dyspepsia (FD) in Guangzhou, and to explore their correlation with some factors such as residence time in Guangzhou, gender, age, and Helicobacter pylori (Hp) infection. Methods A total of 326 cases of FD patients from Guangzhou were investigated. Database of the related clinical data was built and analyzed. Results The commonly-seen TCM syndrome types of FD patients in Guangzhou were spleen-deficiency and qi stagnation, disharmony of liver and stomach, dampness-heat of spleen and stomach, deficiency-cold in spleen and stomach, and concurrence of cold and heat, accounting for 41.71%, 26.07%, 11.96%, 11.04% and 9.20%, respectively. There were significant differences among the TCM syndromes(P < 0.05). The differences of average age of onset, residence time in Guangzhou, FD subtypes and Hp infection were significant among each syndrome type (P < 0.05). Conclusion The distribution of TCM syndrome types of FD in Guangzhou is dominated by spleen-deficiency and qi stagnation, and the syndrome types are correlated with residence time in Guangzhou, gender, average age of onset, FD subtypes, and Hp infection.
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Objective To explore the effect of Exosomes isolated from the A549 lung cancer cells on the proliferation of these cells and their ho?mologous tumor cells,HCC827,and the role of the PI3K/Akt and SRC signaling pathways in this process. Methods Exosomes were isolated from the supernatant after density gradient centrifugation of A549 cells. The Exosomes morphology was observed by transmission electron microscopy. The expression of the Exosome?specific proteins was analyzed using Western blotting. Cell proliferation was investigated using the MTT assay. Re?sults The A549?derived Exosomes were 30?100 nm in diameter and had a bilayer membrane.Western blotting showed that CD9 was detected in these Exosomes. The isolated Exosomes promoted the proliferation of the A549 and the HCC827 cells in a dose?and time?dependent manner,ac?companied by the activation of Akt and SRC. Conclusion Exosomes isolated from A549 cells promote the proliferation of the secreting cells and the homologous tumor cells in a dose?and time?dependent manner. The mechanism may be related to the activation of Akt and SRC.
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Objective To study the diagnostic value of NBI combined with magnification endoscopy using VS classification standard for early gastric carcinoma lesions.Methods A total of 100 patients with suspected early gastric cancer whose gastric mucosa showed roughness,erosion,abnormal colour or ulcer were collected from January 2013 to June 2014.The lesions were observed under white light endoscopy and then underwent biopsy.Observation and biopsy were conducted in the same location by NBI-ME with self contrast method 2 weeks later.Patients in group A underwent NBI-ME,then were diagnosed by VS classifi-cation standard.Patients in group B were diagnosed with white light endoscopy.The sensitivity,specificity, positive predictive value,negative predictive value and accuracy between group A and group B were com-pared.Results The sensitivity,specificity,positive predictive value,negative predictive value and accura-cy of white light endoscopy in the diagnosis of early gastric carcinoma lesions were 76.19% (16 /21 ), 45.57%(36 /79),27.12%(16 /59),87.80%(36 /41)and 52.00%(52 /100),respectively;while the these variables of NBI-ME for early gastric carcinoma lesions were 95.24%(20 /21),97.47%(77 /79), 90.91%(20 /22),98.72%(77 /78)and 97.00%(97 /100),respectively.The accuracy of NBI-ME for early gastric carcinoma lesions was significantly higher than that of white light endoscopy(χ2 =53.30,P <0.01).Conclusion NBI-ME is convenient and effective in the diagnosis of early gastric carcinoma lesions with high consistency of pathology and good clinical application value.
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Objective To investigate the effect of application of video surveillance system for medical staff in the prevention and control of Ebola treatment center (ETC). Methods The high-definition camera installed in the polluted area 100 beds of 24 wards, semi polluted area ring corridor, wear off gowns and other areas without blind spot. Video data and information were collected. The experts of steering group and medical personnel were on duty of video monitoring 24 hours in the medical duty room. They monitored the situation of frontline staff in medical care, whether wearing personal protective equipment (PPE), logistics and transportation problems encountered in the work, and corrected and reminded. Results From December 2015 to January 2014, by checking 182 people on the first-line staff including doctors, nurses and nursing workers, 239 problems of 4 categories were found. The PPE problems was 147 cases (61.50%), while perform operation process issue 35 cases (14.64%), flow logistics distribution problems 31 cases (12.97%), the others 26 cases (10.88%). All the problems were given a timely reminder. There was no infections among medical team. Conclusion Video surveillance system could help medical staff to prevent and control infection, and reduce the risk of infection.
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Objective To construct the secretory expression vector of recombinant human C-reactive protein(rhCRP) for its se-cretory expression in Pichia pastoris ,rhCRP was expressed as a secretory protein and purified ,and the immunity reactivity of the purified protein was identified .Methods The DNA fragment of rhCRP which was designed and synthesized was cloned into pPICZαA vector .Recombinant plasmid pPICZαA/rhCRP was linearized by SacⅠand transformed into Pichia pastoris X-33 by elec-trotransformation .The rhCRP was secreted into the medium under the methanol induction .RhCRP was purified by Histamine affin-ity chromatography .The purified rhCRP was identified by SDS-PAGE and Western blotting ,and its immunity reactivity and stabili-ty was identified by indirect ELISA .Results The pPICZαA/rhCRP expression vector was successfully constructed .The rhCRP of 23 × 103 was inducted and successfully expressed as a secretory protein by the recombinant Pichia pastoris strains .The rhCRP was purified by one step up to 90 .42% purity ,and it was showed good immunity and stability by indirect ELISA .Conclusion The rh-CRP with higher purity and immunoreactivity was successfully obtained by using the Pichia pastoris expression system ,which pro-vided an important experimental basis for producing anti-human CRP antibodies and developing testing CRP reagent .
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Arsenic trioxide (ATO) is a strong inducer of apoptosis in malignant hematological cells. Inducible phosphatidyl inositol 3 kinase (PI3K)-Akt activation promotes resistance to ATO. In the present study, we evaluated whether E3 ubiquitin ligase Cbl-b, a negative regulator of PI3K activation, is involved in the action of ATO. The effect of ATO on cell viability was measured by the Trypan blue exclusion assay or by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was determined by flow cytometry and protein expression was assayed by Western blotting. ATO decreased the viability of HL60 cells and induced cellular apoptosis, which was accompanied by transient activation of Akt. The PI3K/Akt inhibitor, LY294002, significantly increased ATO-induced apoptosis (P < 0.05). In addition, ATO up-regulated the expression of Cbl-b proteins. Furthermore, ATO inhibited cell viability with an IC50 of 18.54 μM at 24 h in rat basophilic leukemia-2H3 cells. ATO induced cellular apoptosis with transient activation of Akt and Cbl-b was also up-regulated. Rat basophilic leukemia-2H3 cells transfected with a dominant negative (DN) Cbl-b mutation showed overexpression of Cbl-b (DN) and enhanced Akt activation. Compared with cells transfected with vector, ATO-induced apoptosis was decreased and G2/M phase cells were increased at the same concentration (P < 0.05). The PI3K/Akt inhibitor, LY294002, re-sensitized Cbl-b (DN) overexpressing cells to ATO and reversed G2/M arrest (P < 0.05). Taken together, these results suggest that Cbl-b potentiates the apoptotic action of ATO by inhibition of the PI3K/Akt pathway.
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Animals , Humans , Rats , Apoptosis/drug effects , Arsenicals/pharmacology , Oxides/pharmacology , /antagonists & inhibitors , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , Proto-Oncogene Proteins c-cbl/pharmacology , Ubiquitin-Protein Ligases/pharmacology , Blotting, Western , Flow Cytometry , Signal Transduction/drug effects , Up-Regulation/drug effectsРеферат
<p><b>BACKGROUND AND OBJECTIVE</b>It has been known that the expression levels of ERCC1 and GST-pi were correlated with tumorigenesis and prognosis. The aim of this study is to investigate the relationship between expression levels of ERCC1 and GST-pi, and clinicopathologic parameters and survival in patients with lung cancer.</p><p><b>METHODS</b>The expression levels of ERCC1 and GST-pi were detected by immunohistochemical staining on tissue micro-array sections made of 148 cases of lung cancer and 7 cases of normal lung samples. The results were compared with relevant clinical and pathologic data.</p><p><b>RESULTS</b>Positive rates of ERCC1 and GST-pi were 36.2% and 73.6%, respectively. None of normal lung samples was positive staining. Positive expression of ERCC1 was significantly higher in group of non-small cell lung cancer (NSCLC), highly differentiated and the smokers less than 400 (P < 0.05), positive expression of GST-pi was significantly higher in group of non-smokers and NSCLC (P < 0.05). There were significant correlations between expression of ERCC1 and GST-pi (r = 0.253, P = 0.001). The 5 years survival rate was higher in positive expression of ERCC1. There was significant correlations between expression of ERCC1 and survival (P = 0.037). There was no significant correlations between expression of GST-pi and survival (P = 0.614). Multivariate analysis using Cox regression model showed that expression levels of ERCC1 and GST-pi were not the important independent prognostic factors for survival.</p><p><b>CONCLUSION</b>ERCC1 and GST-pi are aberrant highly expressed in NSCLC with positive correlation, which indicate they might act synergistically in tumorigenesis of NSCLC. The positive expression of ERCC1 have better survival and may have effect on prognosis.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Carcinoma, Non-Small-Cell Lung , Metabolism , Pathology , DNA-Binding Proteins , Metabolism , Endonucleases , Metabolism , Glutathione S-Transferase pi , Metabolism , Immunohistochemistry , Lung Neoplasms , Metabolism , Pathology , Tissue Array AnalysisРеферат
Objective To investigate the curative effect of methotrexate in treating systemic lupus erythema-tosus(SLE). Methods The chosen outpatients were divided into 2 groups,group A(28 cases) :MTX combined with small dosage of prednisone (prednlsone 10mg/d) ; group B (20 cuses) : medium dosage of prednisone (prednisone 30rag/d). Observation includes SLE disease activity index(SLEDAI) ,relapse while dosage of prednisone was reduced to small amount and drug side effects. Results The SLEDAI score of group A was reduced from (9.4±2.1) to (4.1±0.9) ,and group B from (8.6±2.5) to (4.0±1.0). Compared the result of before and after treatment be-twcen the two groups, the curative effect is obvious (P<0.01). However, there was no significant difference of efficacy between the two groups(P>0.25). While dosage of prednisone was reduced to 10mg/d ,there were 8 cases of relapse in group B,and only 3 cases in group A during the entire observation period. The difference is significant (P<0.025). The side effect of group A is gastrointestinal reaction and for B is mainly the cushing syndrome. Conclusion MTX is an effective treatment fw the light and medium activity SLE,and can reduce the dose of prednisone, prevent long-term using large dose of prednisone which will cause side effect.
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<p><b>BACKGROUND</b>Tissue microarray provides a convenient shortcut for immunohistochemical staining. The aim of this study is to investigate the clinicopathologic and prognostic values of survivin and cyclooxygenase-2 (COX-2) expression level in patients with non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>Expression of survivin and COX-2 was detected in 88 cases of NSCLC and 5 cases of normal lung samples by immunohistochemical staining on tissue microarray sections. All cases were followed up for more than 5 years.</p><p><b>RESULTS</b>Cytoplasmic and nuclear expression rate of survivin in NSCLC was 94.3% and 79.5%, respectively, and positive expression rate of COX-2 was 71.6%, however, neither survivin nor COX-2 expression was observed in normal lung tissues (P < 0.005). Nuclear expression of survivin was markedly higher in smokers than that in non-smokers (P=0.002). The positive expression of COX-2 was significantly related to gender, smoking, histologic subtype and lymph node metastasis (P < 0.05). Univariate analysis showed that patients with positive expression of COX-2 had worse overall survival (P=0.014), however, survivin expression was not related to survival. Multivariate analysis showed that neither survivin nor COX-2 was independent prognostic factor for survival.</p><p><b>CONCLUSIONS</b>The results indicate that survivin highly expresses in NSCLC, so the ubiquitous expression makes it a potential novel parameter for diagnosis of NSCLC. Aberrant expression of COX-2 is related to worse overall survival, which may be useful to predict prognosis for NSCLC.</p>
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Background and purpose:There is an increasing of breast diseases every year. It is important for us to detect the tumor characteristic and to establish the diagnosis of breast diseases through their clinical manifestation and pathology.Methods:A retrospective analysis of 864 cases of breast diseases after surgical operation,was made in our hospital, from 2000 to 2006.Results:85.7% of 864 cases were fibro benign diseases, and most of them were fibro cystic hyperplasia and fibroadenoma of breast.Malignant disease was found 123 cases, and most of them were invasive ductal carcinoma, then medullary carcinoma and early infiltrating carcinoma.It is significant that the youngest age from 30 years old and the incidence rate increased with the age.Conclusions:It is very important to promate the diagnosis rate of breast carcinoma by early prevention and treatment to screen breast diseases and monitor high risk population in time.
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This article began from the characteristic and research method of the syndrome in Traditional Chinese Medicine(TCM)and proteomics,analyzed the superiority and the feasibility of introduction of proteomics into the study of syndrome essence.The author thought that proteome and syndrome in TCM were amazing similar in the aspects of integrity,developments,space and complexity,there was maybe some kind of inner link and the corresponding relations between them.Therefore, the proteomics had the special superiority when it was compared with the general microscopic parameter in study of syndrome essence,also even more accord with the characteristics of syndrome in itself.This article also generalized the research achievement of syndromic proteomics in TCM in recent years from the aspects of specification,thought and method,the relativity of concrete syndrome type and proteome.This article demonstrated the magnificent prospect of proteomics in the study of the syndrome essence of the TCM.
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Objective To investigate the role of resting pulm on ary function (FEV 1,FVC,VC) and diffusing capacity (D LCO) on exercise limitation in patients with chronic obstructive pulmonary disease (COPD).Methods During the period of 2002-04~12 the results of resting pulmonary function test which were and diffusing capacity test performed on 26 subjects with stable COPD were reviewed.The symptom-limited maximum exercise (S LME) were performed.Dyspnea index (DI) and oxygen consumption (V? O 2/kg) were determined with a metabolic cart during exercise.Borg Scale s (BS) was used for counting degree of exertional dyspnea which appeared during exercise.At the end of SLME,arterial blood gas analysis was performed to determi ne PaO 2max,PaCO 2max and Phmax at that time.Results In stepwise multiple regression analysis,the variables significantly contributing to V?O 2max/kg were D LCO % and FEV 1/VC.Conclusion Lung function and diffusing capacity can be importan t determinants of exercise capacity in COPD.
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Objective To compare the effect of ultrafiltration method and activated carbon absorbance method on aconitine-type alkaloid (AA) content in the preparation of Yiqi Huiyang Injection (YHJ) and to evaluate the safety of YHJ by limited detrmination of diester alkaloids (DEA).Methods Acid dye colorimetry was used to detect AA content and reverse phase high performance liquid chromatography (RP-HPLC) was applied to determine the limited content of DEA. Mesaconitine, aconitine, hypaconitine were separated on a Hypersil BDS-C?18?column, with methanol and 0.05% triethylamine (70∶30) as mobile phase, wavelength at 240 nm and external standard for quantitative analysis.Results Compared with activated carbon absorbance method, the loss of AA was reduced, limited content of aconitine was within the pharmacopoeia standard method and the other two kinds of DEA were also low in YHJ prepared by ultrafiltration method.Conclusion Ultrafiltration was superior to activated carbon absorbance in the preparation of YHJ as with AA as the parameter, and the contents of DEA were very low which indicates that the toxicity of YHJ may be also low.
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Objective To establish a HPLC method for the assay of psoralen and isopsoralen in different effective extracts of Fructus Psoraleae. Methods HPLC was carried out on the column of Kromasil RP-C18. The mobile phase was methanol -water(65 ∶35). The flow rate was 1.0 mL/min and the UV detection wavelength was 245 nm. Results Good linearity of psoralen was showed within the range of 10.5 ng~525 ng(r= 0.999 3)and isopsoralen within the range of 9 ng~450 ng (r= 0. 999 9). The content of psoralen and isopsoralen differed in different extractions of Fructus Psoraleae. Among them,the extract C (extracted by ethyl acetate ) contained the highest contents of psoralen and isopsoralen,while the contents of psoralen and isopsoralen were very low in the extract D (extracted by n-butyl alcohol) and E (supernatant of water extract). Conclusion The method is simple,accurate and reproducible. The anti-asthma effect and the dose-effect relationship of the different effective extracts of Fructus Psoralea need further pharmacodynamics study.
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Objective To investigate the distribution of the mini mental status examination in the elderly Methods Totally 3 776 residents aged 55 years or over serving as samples in the urban and rural areas of Lhasa city, from August to December in 2001 were studied A residential area and a door to door general sampling procedures were used in this study, respectively Mini mental status examination (MMSE) and a activity of daily living scale (ADL) were used as screening methods for dementia Results The distribution of lower scores in such examination in middle elderly people, in Lhasa city was 7 2%, in which, males were 9 3% and femals, 6 3% Conclusion The results of lower scores in the mini mental status examination were similar to those reporting from inland cities, but there appear some what differences in distributions of ages, educational levels and sexes from Tibetan, which may be due to their special location and living habits