1.Extraction of Volatile Oil of Rhizoma Zedoariae in Weishu Granules and Preparation of Its ?-cyclodextrin-inclusion complexes
Wei HE ; Zhufeng CONG ; Yong LI
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To optimize the conditions of process for extraction of volatile oil of Rhizoma Zedoariae (RZ) in Weishu Granules and preparation of its ? - cyclodextrin- inclusion complexes. Methods Orthogonal design was adopted. Extraction content of RZ volatile oil was used as the index for the optimizing of extraction process. The amount of inclusion complexes and efficient inclusion rate served as the indexes for the optimizing of inclusion process. Ultraviolet spectrophotometry was applied to detect the content of volatile oil. Results The optimal extraction conditions were: gringding the crude RZ into coarse grains, adding 12- fold water and distilling the mixture for 6 h; the optimal inclusion conditions were: ? - cyclodextrin and volatile oil were mixed in the proportion of 8 :1, 2- fold water was added into the mixture and the mixture was triturated for 60 min. The maximum UV spectrum absorbance of aethylis acetas liquor of valatile oil located at wavelength of 253nm. Conclusion Higher extraction rate of RZ volatile oil can be achieved by this technique; volatile oil is powdered and its stability increased after being included by ? - cyclodextrin.
2.Ribosome display screening of a novel human anti-IgE scFv fragment.
Yongxia ZHANG ; Baocheng WANG ; Xin YU ; Yunjian DAI ; Yongzhi HE ; Cong CONG ; Yong XIA ; Mingrong WANG
Acta Pharmaceutica Sinica 2012;47(10):1329-35
Total mRNA was extracted from lymphocytes separated from the peripheral blood of allergic patients, and then variable region of heavy chain (VH) and variable region of light chain (VL) cDNA library were constructed by RT-PCR. Human scFv templates for rabbit reticulocyte lysate ribosome display were assembled by primers and linker peptide (Gly4Ser)3. mRNA bound in antibody-ribosome-mRNA complexes was recovered using in-situ single primer RT-PCR, and three rounds of anti-IgE scFv DNA were enriched. The target DNA fragments were double enzyme digested and ligated into plasmid pET22b (+), followed by transformation in E. coli Rosseta (DE3). Positive clones were screened using clone PCR, Dot blotting and antigen ELISA. The correct lengths of VH (400 bp) and VL (710 bp) PCR products were obtained. The expected 1,000 bp ribosome display templates were also observed in agarose gel electrophoresis. After three rounds of ribosome display target sequences were effectively enriched, leading to a library of 10(13) members. Antibodies with the highest ELISA value for IgE were generated in the strain pET-IgE-6. A human anti-IgE scFv library was successfully constructed as described herein. Ribosome display using single primer in-situ RT-PCR as the recovery procedure effectively enriched target sequences. Anti-IgE scFv with high affinity and specificity were identified. The prepared human anti-IgE scFv fragment might be self-developed to a lead drug for treating asthma. Our study provides an alternative method for rapid discovery of human antibodies of therapeutic importance.
3.Ribosome display screening of a novel human anti-IgE scFv fragment.
Yong-xia ZHANG ; Bao-cheng WANG ; Xin YU ; Yun-jian DAI ; Yong-zhi HE ; Cong CONG ; Yong XIA ; Ming-rong WANG
Acta Pharmaceutica Sinica 2012;47(10):1329-1335
Total mRNA was extracted from lymphocytes separated from the peripheral blood of allergic patients, and then variable region of heavy chain (VH) and variable region of light chain (VL) cDNA library were constructed by RT-PCR. Human scFv templates for rabbit reticulocyte lysate ribosome display were assembled by primers and linker peptide (Gly4Ser)3. mRNA bound in antibody-ribosome-mRNA complexes was recovered using in-situ single primer RT-PCR, and three rounds of anti-IgE scFv DNA were enriched. The target DNA fragments were double enzyme digested and ligated into plasmid pET22b (+), followed by transformation in E. coli Rosseta (DE3). Positive clones were screened using clone PCR, Dot blotting and antigen ELISA. The correct lengths of VH (400 bp) and VL (710 bp) PCR products were obtained. The expected 1,000 bp ribosome display templates were also observed in agarose gel electrophoresis. After three rounds of ribosome display target sequences were effectively enriched, leading to a library of 10(13) members. Antibodies with the highest ELISA value for IgE were generated in the strain pET-IgE-6. A human anti-IgE scFv library was successfully constructed as described herein. Ribosome display using single primer in-situ RT-PCR as the recovery procedure effectively enriched target sequences. Anti-IgE scFv with high affinity and specificity were identified. The prepared human anti-IgE scFv fragment might be self-developed to a lead drug for treating asthma. Our study provides an alternative method for rapid discovery of human antibodies of therapeutic importance.
Amino Acid Sequence
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Antibodies, Anti-Idiotypic
;
genetics
;
isolation & purification
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Antibody Affinity
;
Asthma
;
blood
;
Base Sequence
;
DNA, Complementary
;
metabolism
;
Escherichia coli
;
metabolism
;
Humans
;
Immunoglobulin Heavy Chains
;
genetics
;
Immunoglobulin Light Chains
;
genetics
;
Immunoglobulin Variable Region
;
genetics
;
Lymphocytes
;
chemistry
;
Peptide Library
;
RNA, Messenger
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isolation & purification
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Recombination, Genetic
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genetics
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Ribosomes
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chemistry
;
genetics
;
immunology
;
Single-Chain Antibodies
;
genetics
;
isolation & purification
;
Transformation, Genetic
4.A novel method for testing sterility of injections based on biothermodynamics.
Dan GAO ; Dan GAO ; Yong-Shen REN ; Dan YAN ; Cong-En ZHANG ; Zhu-Yun YAN ; Yin XIONG ; Li-Na MA ; Le-Le ZHANG ; Xiao-He XIAO
Acta Pharmaceutica Sinica 2014;49(3):385-391
This study aims at trying to establish a novel method of sterility test for injections based on biothermodynamics, in order to overcome the deficiencies of routine sterility tests such as long detecting cycle, low sensitivity and prone to misjudgments. A biothermodynamics method was adopted to rapidly detect the microorganism contamination of injections by monitoring the heat metabolism during the growth of microbe. The growth rate equal to or greater than zero and the heat power difference of P(i) and P(0) with three folds higher than the noise of baseline were chosen as indexes to study the heat change rule of microbe. In this way, the effectiveness of the new method to detect strains required by conventional sterility test or in injection samples was also investigated. Results showed that the Gram-positive bacteria, Gram-negative bacteria and fungi demanded by sterility testing methodology could be detected by biothermodynamics method within 10 hours, with the sensitivity lower than 100 CFU x mL(-1). Meanwhile, this method was successfully applied to the sterility test of Compound Yinchen injection (FFYC), Shuanghuanglian powder injection (SHL) and Compound Triamcinolone injection (TAND) which were sterilized with different degrees. Therefore, the biothermodynamics method, with advantages of fast detection and high sensitivity, could be a complementary solution for conventional sterility tests.
Anti-Inflammatory Agents
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administration & dosage
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chemistry
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Drug Contamination
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Drugs, Chinese Herbal
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administration & dosage
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chemistry
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Fungi
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isolation & purification
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Gram-Negative Bacteria
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isolation & purification
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Gram-Positive Bacteria
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isolation & purification
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Hot Temperature
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Injections
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Microbiological Techniques
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methods
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Sensitivity and Specificity
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Sterilization
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Triamcinolone
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administration & dosage
;
chemistry
5.Comparison of the exposure rates of risk factors and inducing factors of acute myocardial infarction between younger and aged adults.
Cong-gang HUANG ; Rui LI ; Hui-ping ZUO ; Zheng-yan WANG ; Rong-hua HE ; Yong-guang ZOU
Chinese Journal of Epidemiology 2007;28(3):282-285
OBJECTIVETo study the clinical characteristics of acute myocardial infarction (AMI) among younger adults and to explore the possible mechanisms of early myocardial infarction, combined with the newly discovered risk factors of coronary heart disease.
METHODSData on comparative analysis to the exposure rates of the risk factors and inducing factors of non-CAD patients with two groups of AMI patients including younger adults group (< or =40 years old) and aged adults group (> or =50 years old). Coronary angiography was applied.
RESULTSThere were differences noticed between the frequencies of risk factors of the two AMI groups. In younger adults group the exposure rates of smoking, hyperlipidemia, positive family history, C-reactive protein (CRP) and fibrinogen were markedly higher, while in elderly group the exposure rates of hypertension, smoking, hyperlipidemia, diabetes, CRP, fibrinogen and homocysteine (HCY) were markedly higher (P < 0.05). Although the clustering status of risk factors of the younger adult group was not higher than that of the elderly group. There were obvious inducing factors before the patients were attacked by AMI and the inducing factors inclined to cluster, which had obvious dose-reaction relationships with the occurrence of AMI in young people.
CONCLUSIONEarly AMI of younger adults might relate to the clustering status of inducing factors. The coexistence of several kinds of inducing factors was resulted in the occurrence of AMI of the atherosclerosis (As) and non-As patients by means of myocardial ischemia accumulation effect.
Adult ; Age Factors ; Aged ; Atherosclerosis ; epidemiology ; China ; epidemiology ; Coronary Angiography ; Humans ; Middle Aged ; Myocardial Infarction ; epidemiology ; pathology ; Myocardial Ischemia ; epidemiology ; Risk Factors
6.Clinical psychological factors and treatment strategies for patients with chronic hepatitis B
Haofei WANG ; Yuqin WU ; Xiaoyin CONG ; Yong LI ; Danjun HE
Journal of Clinical Medicine in Practice 2018;22(5):55-58
Objective To analyze the correlation between clinical mental personality,coping style,social support and depression,anxiety for patients with chronic hepatitis B,and their treatment strategies.Methods A total of 66 patients with chronic hepatitis B (CHB group) and 72 normal people (control group) were investigated by the questionnaire,such as the demographic history questionnaire,Eysenck Personality Questionnaire (EPQ),Trait Coping Style Questionnaire (TCSQ),Perceived Social Support Scale (PSSS),Hamilton Depression Scale (HAMD),Hamilton Anxiety Scale (HAMD) were assessed.Results The HAMD score and HAMA score of hepatitis B group were significantly higher than that of the control group.Pearson correlation analysis showed that depressive symptoms was negatively correlated with positive coping,family support,extroversion,and was positively correlated with neuroticism.Multiple linear regression analysis showed family support,active coping,neuroticism and psychosis were all included in the regression equation and can predict depressive symptoms (3 values,0.002,0.003,0.004,0.022,respectively).Neuroticism,positive responded to the two independent variables (β values,0.003,0.011,respectively) into the regression equation,and can predict anxiety symptoms.Conclusion The patients with chronic hepatitis B were more prone to suffer from depression and anxiety than normal people,depressive symptoms are closely related to the neuroticism and neuroticism.The anxiety symptoms are related to neuroticism.The neurotic personality traits,lack of positive coping strategies and out-of-home support may lead to depression and anxiety.Depression and anxiety symptoms can be avoided by improving the patient's personality characteristics,and actively responding and enhancing the social support system outside the home soas to reduce depression,anxiety symptoms in patients with hepatitis B.
7.Clinical psychological factors and treatment strategies for patients with chronic hepatitis B
Haofei WANG ; Yuqin WU ; Xiaoyin CONG ; Yong LI ; Danjun HE
Journal of Clinical Medicine in Practice 2018;22(5):55-58
Objective To analyze the correlation between clinical mental personality,coping style,social support and depression,anxiety for patients with chronic hepatitis B,and their treatment strategies.Methods A total of 66 patients with chronic hepatitis B (CHB group) and 72 normal people (control group) were investigated by the questionnaire,such as the demographic history questionnaire,Eysenck Personality Questionnaire (EPQ),Trait Coping Style Questionnaire (TCSQ),Perceived Social Support Scale (PSSS),Hamilton Depression Scale (HAMD),Hamilton Anxiety Scale (HAMD) were assessed.Results The HAMD score and HAMA score of hepatitis B group were significantly higher than that of the control group.Pearson correlation analysis showed that depressive symptoms was negatively correlated with positive coping,family support,extroversion,and was positively correlated with neuroticism.Multiple linear regression analysis showed family support,active coping,neuroticism and psychosis were all included in the regression equation and can predict depressive symptoms (3 values,0.002,0.003,0.004,0.022,respectively).Neuroticism,positive responded to the two independent variables (β values,0.003,0.011,respectively) into the regression equation,and can predict anxiety symptoms.Conclusion The patients with chronic hepatitis B were more prone to suffer from depression and anxiety than normal people,depressive symptoms are closely related to the neuroticism and neuroticism.The anxiety symptoms are related to neuroticism.The neurotic personality traits,lack of positive coping strategies and out-of-home support may lead to depression and anxiety.Depression and anxiety symptoms can be avoided by improving the patient's personality characteristics,and actively responding and enhancing the social support system outside the home soas to reduce depression,anxiety symptoms in patients with hepatitis B.
8.Application of standardized uptake value for FDG PET-CT in predicting radiation pneumonitis.
Yong ZHANG ; Yong-hua YU ; Jin-ming YU ; Wei HE ; Zheng FU ; Shou-fang GUO ; Xi-jun LIU ; Chang-sheng CONG
Chinese Journal of Oncology 2009;31(8):622-625
OBJECTIVETo investigate the correlation of radiation pneumonitis (RP) with standardized uptake value (SUV) for fluorodeoxyglucose (FDG) positron emission tomography and computed tomography (PET-CT) in lung cancer patients treated with radiation therapy.
METHODSFourty patients with unresectable non-small cell lung cancer (NSCLC) received FDG PET-CT before and after radiotherapy. The average SUV of the lung tissue irradiated with a dose of < or = 5 Gy, 5.1 approximately 15 Gy, 15.1 approximately 35 Gy, 35.1 approximately 60 Gy, >60 Gy were measured. The correlation between SUV and RP was analyzed by comparing the SUV in the patients with RP and without. The SUV ratio of the irradiated lung tissue to that of the non-irradiated lung tissue (L/B) was also calculated.
RESULTSOf the 40 patients, 8 developed RP, including 6 cases of grade 2 and 2 cases of grade 3. The SUV of irradiated lung tissues with a dose of 35.1 approximately 60 Gy was significantly correlated with RP. When SUV > or =1, the RP incidence rate was 41.7% versus 20.0% in the whole group, with a statistically significant difference. (chi2 = 3.96, P < 0.05). The sensitivity and specificity of SUV in predicting RP was 62.5% and 78.1%, respectively. When the value of L/B > or = 2.5, the RP incidence rate was 40.7% in this group versus 20.0% in the whole group, with a statistical significance (chi(2) = 4.92, P < 0.05). If taking L/B > or = 2.5 as a threshold value, the sensitivity and specificity in predicting RP was 72.7% and 90.9%, respectively. No statistically significant difference was found in predicting radiation pneumonitis between SUV > or =1 and L/B > or = 2.5 (chi2 = 0.002, P > 0.05).
CONCLUSIONThe standardized uptake value (SUV) and the SUV ratio of the irradiated lung tissue to that of the non-irradiated lung tissue (L/B) for FDG PET-CT are positively correlated with radiation pneumonitis, and clinicians may use it to predict the occurrence of radiation pneumonitis.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; radiotherapy ; Female ; Fluorodeoxyglucose F18 ; pharmacokinetics ; Follow-Up Studies ; Humans ; Lung Neoplasms ; radiotherapy ; Male ; Middle Aged ; Positron-Emission Tomography ; Radiation Dosage ; Radiation Pneumonitis ; diagnosis ; etiology ; metabolism ; Radiopharmaceuticals ; pharmacokinetics ; Radiotherapy, Conformal ; adverse effects ; Tomography, X-Ray Computed
9.Estimation of postmortem interval by detecting thickness of cornea using ultrasonic method.
Guo-Li LÜ ; Fu-Xue JIANG ; Xin-Shu XU ; Yong-Jun JIANG ; Zhi-Gang LI ; Xin WANG ; He SHI ; Li-Cong YU ; Chuan-Chao XU
Journal of Forensic Medicine 2012;28(2):89-91
OBJECTIVE:
To explore the postmortem changes of cornea thickness measured by ultrasonic pachymetry.
METHODS:
Eleven rabbits were randomly divided into two groups: one group with intact corneal epithelium and another group without intact corneal epithelium. In the later group, the corneal epithelium of the rabbit was scraped using mechanical elimination method. The corneal thickness was monitored continuously by ultrasonic pachymetry at several postmortem interval points in rabbits of the two groups. The changes of corneal thickness and postmortem interval were explored by relative regression analysis.
RESULTS:
The thickness of the cornea showed a strong non-linear correlation with the postmortem interval in the group with intact corneal epithelium. The group with intact corneal epithelium showed the correlation coefficient 0.922 and the group without intact corneal epithelium showed the correlation coefficient 0.822, respectively.
CONCLUSION
The corneal thickness measured by ultrasonic pachymetry shows a potential value for estimating early postmortem interval. The intact corneal epithelium is a crucial factor for the measurement of cornea thickness by ultrasonic pachymetry.
Animals
;
Cornea/pathology*
;
Corneal Topography/methods*
;
Epithelium, Corneal/ultrastructure*
;
Female
;
Forensic Pathology/methods*
;
Male
;
Postmortem Changes
;
Rabbits
;
Regression Analysis
;
Reproducibility of Results
;
Time Factors
;
Ultrasonography
10.Preliminary biomarker related to nasopharyngeal carcinoma filtered from the whole genome expression profiling involved in microdissection nasopharyngeal tissues.
Zhong-qi LIU ; Yong-quan TIAN ; He HUANG ; Hou-de ZHOU ; Qiu-hong ZHANG ; Ming ZHOU ; Cong PENG ; Xiao-ling LI ; Gui-yuan LI
Journal of Central South University(Medical Sciences) 2005;30(1):1-6
OBJECTIVE:
To filter biomarkers of nasopharyngeal carcinoma (NPC) by constructing the homogenesis tissue gene expression profiling with the whole human genome GeneChip.
METHODS:
The epithelium cells of the homogenesis NPC and the pure nasopharyngeal normal tissues microdissected from nasopharyngeal biopsy which was preserved in the RNAlater were used to isolate RNA and then to harvest the aRNA through in vitro transcription, and aRNA prober was labled to hybridize to HG-U133. plus 2.0, so the expression profiling of each homogenesis tissue could be constructed.
RESULTS:
Some candidate biomarker genes related to the tumorigenesis of NPC had been filtered by comparing the expression profiling of NPC samples with the expression profiling of normal nasopharyngeal epithelia samples. Any genes regarding the metastasis of NPC might have been selected by comparing the expression profiling of no-metastasis samples with those of the metastasis samples.
CONCLUSION
Using the whole genome GeneChip to construct the expression profiling for the microdissected homogenesis tissue is effective to filter the candidate biomarker genes.
Adult
;
Aged
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Biomarkers, Tumor
;
Female
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Gene Expression Profiling
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Gene Expression Regulation, Neoplastic
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Humans
;
Male
;
Microdissection
;
Middle Aged
;
Nasopharyngeal Neoplasms
;
genetics
;
Nasopharynx
;
metabolism
;
Neoplasm Proteins
;
biosynthesis
;
genetics
;
Oligonucleotide Array Sequence Analysis
;
Tumor Suppressor Proteins
;
biosynthesis
;
genetics