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OBJECTIVES@#The previously established 38-plex InDel system was optimized and its performance was validated according to the Scientific Working Group on DNA Analysis Method (SWGDAM) application guidelines. The ancestry inference accuracy of individuals from East Asian, European, African and mixed populations was verified.@*METHODS@#DNA standard sample 9947A was used as the template to establish the optimal amplification conditions by adjusting primer balance, Mg2+ final concentration and optimizing PCR thermal cycle parameters and amplification volume. The allelic dropout, nonspecific amplification and whether the origin of the inferred samples matched the known information were compared to evaluate the performance of this system.@*RESULTS@#The optimal dosage of this system was 0.125-2 ng DNA template. The results of InDel typing were accurate, the amplification equilibrium was good, and the species specificity was good. This system showed certain tolerance to DNA samples including the inhibitor such as hemoglobin (≤80 μmol/L), indigo (≤40 mmol/L), calcium ion (≤1.0 mmol/L), and humic acid (≤90 ng/μL). The system enabled the direct amplification of DNA from saliva and blood on filter paper, and the results of ethnic inference were accurate. The system successfully detected the mixed DNA sample from two individuals. The test results of the system for common biological materials in practical cases were accurate.@*CONCLUSIONS@#The results of the 38-plex InDel system are accurate and reliable, and the performance of the system meets the requirement of the SWGDAM guidelines. This system can accurately differentiate the ancestry origins of individuals from African, European, East Asian, and Eurasian populations and can be implemented in forensic practice.
Subject(s)
Humans , Polymorphism, Single Nucleotide , Genotype , Polymerase Chain Reaction , DNA/genetics , DNA Fingerprinting/methods , INDEL Mutation , Genetics, Population , Gene FrequencyABSTRACT
<p><b>OBJECTIVE</b>To study the pharmacokinetics of puerarin (PUE) in Gegen Qinlian Decoction (, GQD), and the effects of PUE dosage variations on the pharmacokinetics of baicalin (BAL) in mice.</p><p><b>METHODS</b>GQD is composed of the concentrated granules of four Chinese herbs. Three dosages with different levels of PUE, including GQD, GQD co-administered with PUE, and GQD co-administration with two times the amount of PUE, were used to research the pharmacokinetics of PUE and BAL in mice. The indirect competitive enzyme-linked immunosorbent assay (icELISA) methods based on an anti PUE-monoclonal antibody (MAb)and BAL-MAb were employed to determine the concentration of PUE and BAL in mice blood.</p><p><b>RESULTS</b>After the co-administration of GQD with PUE, the area under the curves (AUC) of PUE increased 2.8 times compared with GQD. At the dose of GQD co-administration at two times that of PUE, the (AUC) of PUE was almost equal to that of GQD co-administration of PUE, showing non-linear pharmacokinetics. The (AUC) of BAL showed a good dose-related increase of PUE (r=0.993) in the range from 100 to 300 mg/kg, indicating that PUE dramatically affects the absorption of BAL in mice. There was no significant difference in the other pharmacokinetic parameters, such as the first time of maximum concentration (T), the second T, or the mean residence time.</p><p><b>CONCLUSIONS</b>The icELISA methods were successfully applied to pharmacokinetic studies of PUE and BAL in GQD in mice. The dosage variability of PUE of the main ingredient in GQD affects its own pharmacokinetic characteristics and the absorption characteristics of BAL.</p>
Subject(s)
Animals , Male , Mice , Drugs, Chinese Herbal , Pharmacokinetics , Enzyme-Linked Immunosorbent Assay , Flavonoids , Pharmacokinetics , Herb-Drug Interactions , Isoflavones , Pharmacology , Vasodilator Agents , PharmacologyABSTRACT
Purpose To investigate the relationship between M2 microphage infiltration and the pathological grading and staging of prostate cancer.Methods 58 patients who accepted the radical resection of prostate cancer were selected.After operation,pathologic specimens were hematoxylin eosin stained and immunohistochemical of SP stained to identify the pathological grading,staging,and M2 microphage infiltration.Then,Chi-squared test was used to analyze the relationship between M2 microphage infiltration and the pathological grading and staging of prostate cancer.Results There were 71.4% high stage (≥T2c) cases and 60% lymph node metastasis cases in M2 microphage infiltration group,which was significantly higher than those in non M2 microphage infiltration group.However,there was no relationship between M2 microphage infiltration and pathological grading.Conclusion M2 macrophage infiltration may participate in the prostate cancer cell growth and metastasis,whose pathophysiological mechanism needs further exploration.
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Immunogenic antigen (jujuboside A-BSA) and coating antigen (jujuboside A-OVA) of jujuboside A were synthesized by sodium periodate oxidation method for the first time. Jujuboside A artificial antigen was confirmed by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF-MS). The titer and specificity of the antibody in serum of immunized mice were detected by enzyme-linked immunosorbent assay (ELISA). The corrected relation curve of inhibition rate showed that the antibody against Jujuboside A obtained from immunized mice could bind to jujuboside A and the titer was up to 1∶4 000. The jujuboside A artificial antigen was synthesized, which can be used further to preparation of monoclonal antibody and the pharmacokinetics study of jujuboside A in laboratory animals.
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Sini Powder (SP), a traditional Chinese herbal formula, has long been used to treat depression in patients, although the underlying mechanisms remain to be elucidated. In the present study, we found that rats treated with SP extract for 7 days showed a significant increase in swimming time and reduction in immobility time in forced swimming test in a dose-dependent manner, without changes in locomotion. These effects could be attributed to SP's modulation of the hypothalamus-pituitary-adrenal axis, because a single pretreatment of SP extract could rescue increased serum corticosterone and plasma adrenocorticotropin levels induced by acute elevated platform stress. A single pretreatment of SP extract could also elevate the mRNA expression of hippocampal glucocorticoid receptors. In conclusion, our results suggest that SP extract may act as an anti-stress medication to produce antidepressant-like effects.
Subject(s)
Animals , Humans , Male , Rats , Adrenocorticotropic Hormone , Blood , Antidepressive Agents , Corticosterone , Blood , Depression , Drug Therapy , Genetics , Metabolism , Drugs, Chinese Herbal , Hippocampus , Pituitary-Adrenal System , Metabolism , Rats, Sprague-Dawley , Receptors, Glucocorticoid , Genetics , MetabolismABSTRACT
The method of monoclonal antibody-based immunoassay has a great importance in the study of quality control of traditional Chinese medicine (TCM) and detection of trace components in vivo animals. Synthesis of small molecule artificial antigen is the prerequisite for the establishment of this method. In present study, catalpol-BSA was synthesized by sodium periodate oxidation method. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF-MS) and molecular exclusion chromatography showed that catalpol was successfully conjugated with BSA. The mice could specifically produce anti-catalpol antibodies with titer up to 1:8000. The artificial antigen of catalpol was successfully synthesized.
Subject(s)
Animals , Male , Mice , Antibodies , Allergy and Immunology , Antigens , Chemistry , Allergy and Immunology , Immunoassay , Iridoid Glucosides , Chemistry , Allergy and Immunology , Medicine, Chinese Traditional , Mice, Inbred BALB C , Serum Albumin, Bovine , Chemistry , Allergy and ImmunologyABSTRACT
Oxidation method with sodium iodide was used to synthesize immunogenic antigen (PF-BSA) and coating antigen (PF-OVA) of paeoniflorin. UV spectroscopy showed that paeoniflorin was successfully conjugated with BSA and OVA. After immunized by PF-BSA, the mice can produce anti-paeoniflorin antibodies specifically. The ELISA test results showed the high titer (1:12 800) and specificity (IC50 = 0.791 mg x L(-1)) of the antiserum from mice injected with PF-BSA. Also, the antiserum showed low cross activities against nine traditional Chinese medicine (TCM) of small molecules. These artificial antigens were successfully synthesized and the anti-paeoniflorin antibody well prepared, which provides the experimental basis for the further study of ELISA and its kit.
Subject(s)
Animals , Male , Mice , Antibodies , Antigens , Chemistry , Allergy and Immunology , Drugs, Chinese Herbal , Chemistry , Enzyme-Linked Immunosorbent Assay , Glucosides , Chemistry , Allergy and Immunology , Mice, Inbred BALB C , Monoterpenes , Chemistry , Allergy and Immunology , Serum Albumin, Bovine , Chemistry , Allergy and ImmunologyABSTRACT
The establishment of high specificity and sensitivity method of small molecule monoclonal antibody-based immunoassay has a great importance in the study of small molecule compounds in Chinese medicine, wherein synthesis of small molecule artificial antigen is a critical step in the preparation of small molecule antibodies. Oxidation method using sodium iodide was used to synthesize immunogenic antigen (FRn-BSA) and coating antigen (FRn-OVA) of forsythin. UV spectroscopy and thin layer chromatography showed that forsythin was successfully conjugated with BSA and OVA. After immuned FRn-BSA, the mice could specifically produce anti-forsythin antibodies with titer up to 1:8 000, and the linear range was from 1 mg x L(-1) to 100 mg x L(-1). In this paper, the artificial antigen of forsythin was successfully synthesized, which can be applied for preparation of monoclonal antibodies and establishment of appropriate immune method.
Subject(s)
Animals , Male , Mice , Antibodies , Allergy and Immunology , Antigens , Chemistry , Allergy and Immunology , Bridged Bicyclo Compounds, Heterocyclic , Chemistry , Allergy and Immunology , Drugs, Chinese Herbal , Chemistry , Furans , Chemistry , Allergy and Immunology , Mice, Inbred BALB CABSTRACT
Study on pharmacodynamic material basis of traditional Chinese medicines is one of the key issues for the modernization of traditional Chinese medicine. Having introduced the monoclonal antibody technology into the study on pharmacodynamic material basis of traditional Chinese medicines, the author prepared the immunoaffinity chromatography column by using monoclonal antibodies in active components of traditional Chinese medicines, so as to selectively knock out the component from herbs or traditional Chinese medicine compounds, while preserving all of the other components and keeping their amount and ratio unchanged. A comparative study on pharmacokinetics and pharmacodynamics was made to explicitly reveal the correlation between the component and the main purpose of traditional Chinese medicines and compounds. The analysis on pharmacodynamic material basis of traditional Chinese medicines by using specific knockout technology with monoclonal antibodies is a new method for study pharmacodynamic material basis in line with the characteristics of traditional Chinese medicines. Its results can not only help study material basis from a new perspective, but also help find the modern scientific significance in single herb or among compounds of traditional Chinese medicines.
Subject(s)
Animals , Humans , Adsorption , Antibodies, Monoclonal , Chemistry , Cell Line , Chromatography, Affinity , Methods , Drugs, Chinese Herbal , Chemistry , PharmacologyABSTRACT
Compatibility mechanism study of Chinese herbal compound (CHC) has been one of key contents in Chinese medical research, but the present research methods are not suitable for its own features due to its complexity, which has restricted the process of modernization and intemationalization of Chinese medicine. In this paper, we addressed that the compatibility is closely correlated to their in vivo metabolic processes. The preparation of active small molecules monoclonal antibodies in herbs and testing a variety of effective compositions simultaneously using immunoassay can clarify the in vivo metabolism and mutual interactions of Chinese herbs, which is a new thought of studying the compound compatibility mechanisms.
Subject(s)
Antibodies, Monoclonal , Drug Combinations , Drug Incompatibility , Drugs, Chinese HerbalABSTRACT
To describe the imaging features of gastrointestinal stromal tumors [GISTs] at initial presentation with clinical, surgical, and pathologic correlation, and to evaluate values of various techniques in GISTs. This retrospective study recruited 70 patients with histologically proved GISTs between December 2004, and May 2009 in the Department of General Surgery, Zhongshan Hospital, Fudan Univeristy, Shanghai, China. Each patient underwent CT scanning, 39 patients underwent simultaneous endoscopy, 12 patients underwent endoscopic ultrasound [EUS], and 36 patients underwent transabdominal ultrasonography [TAUS] simultaneously. Features of GISTs were assessed. Computerized tomography findings showed an eccentric mass in 44 patients, an intraluminal component in 24, and a transmural distribution in 2. Forty-two tumors were dumbbell-shaped, 2 were round, while 26 were irregular. Forty-three tumors presented with well-defined masses, while 27 with unclear borders. The arterial phase attenuation showed the continuous enhancement. The portal-venous phase attenuation was heterogeneous in 26 and homogeneous in the other 44. There was a significant correlation between certain CT features and tumor risk stratification. Gastrointestinal stromal tumors were characterized by a smooth shape and normal overlying mucosa in endoscopy, hypoechoic, and solid in TAUS. Imaging examinations are pivotal in the management of GISTs. The CT scan is valuable in the diagnosis, staging, and treatment planning of GISTs. Endoscopy and EUS contribute to the detection of mucosal lesions. Other methods including TAUS, fluorodeoxyglucose positron emission tomography, CT gastrography, and MRI help in specific cases
Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Gastrointestinal Stromal Tumors/pathology , Gastrointestinal Stromal Tumors/surgery , Retrospective Studies , Endosonography , Ultrasonography , Tomography, X-Ray ComputedABSTRACT
<p><b>OBJECTIVES</b>To investigate the value of R2*map for quantitatively tracing superparamgnetic ironoxides (SPIO) labeled endothelial progenitor cells (EPCs).</p><p><b>METHODS</b>The EPCs were isolated from Balb/c mice bone marrow and cultured in vitro. After 7 days, expression of acetylated low-density lipoprotein (acLDL) and Ulex europaeus agglutinin-1 (UEA-1), two markers of EPCs, was observed by double staining using fluorescence microscope, the expression of stem cell antigen-1 and vascular endothelial growth factor receptor-2 (VEGFR-2) was confirmed by flow cytometry. EPCs were labeled by incubating with 50 microg/ml SPIO and 6 microl/ml lipofectamine2000, SPIO labeled EPCs were observed under transmission electron microscope (TEM). Labeled and unlabeled EPCs were mixed with 10 g/L agarose and scanned using a 3.0T MR scanner, R2* map and R2 map images were obtained on workstation.</p><p><b>RESULT</b>After 7 days of in vitro culture, most of the cells showed characteristics of EPCs. There was no morphological difference between SPIO labeled EPCs and unlabeled EPCs. R2* and R2 values exhibited a linear correlation with the number of labeled cells in the agarose gel. Compared to R2, R2* was a better indicator of the number of labeled cells.</p><p><b>CONCLUSION</b>MR R2* map can be used to trace SPIO labeled EPCs quantitatively.</p>
Subject(s)
Animals , Mice , Cells, Cultured , Endothelial Cells , Cell Biology , Ferric Compounds , Chemistry , Magnetic Resonance Imaging , Methods , Magnetics , Mice, Inbred BALB C , Nanoparticles , Chemistry , Stem Cells , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of Banxia Xiexin Decoction (BD) and its components in the rats with electrogastric dysrhythmia, explore the mechanism of BD' s "relieving distension and fullness, and dissolving lumps" and study BD' s compatibility regularity.</p><p><b>METHODS</b>Rat model of electrogastric dysrhythmia was established, the effects of BD and its components on the coefficient of variability of the model' s slow wave electrogastric rhythm were determined.</p><p><b>RESULTS</b>Compared with the model group, BD and all the components groups had better effects on all the aspects (P <0. 05 ), Among which the group containing herbs with pungent and bitter flavour (Rhizoma Pinelliae, Rhizoma Zingiberis, Radix Scutellariae, Radix Berberidis julianae) was the best.</p><p><b>CONCLUSION</b>BD and all the components have the effects of rectifying electrogastric dysrhythmias in various degrees, and the herbs with pungent and bitter flavour are best. This study provides the experimental foundations for further understanding the mechanism of BD' s treating principle-using bitter drugs for purgation and pungent drugs for dispersion, normalizing the functional activities of qi.</p>
Subject(s)
Animals , Rats , Digestive System , Electrodiagnosis , Gastrointestinal Motility , Medicine, Chinese Traditional , Myoelectric Complex, Migrating , Plant Extracts , Pharmacology , Stomach DiseasesABSTRACT
Objective: To summarize the distributing rule of TCM syndrome elements of common diseases by analyzing the literature of Depression,Chronic Hepatitis B,Chronic Renal Failure,Chronic Obstructive Pulmonary Disease,Menopausal Syndrome.Methods: We selected eligible literatures,then established database with EpiData 3.0 software and computed the frequency of TCM syndrome elements with SPSS 12.0 software.Results: ①Liver,heart,spleen,lung and kidney are common TCM syndrome elements of location of disease.②Yin deficiency,yang deficiency,heat,qi stasis,qi deficiency,blood stasis,damp and phlegm are common TCM syndrome elements of nature of disease.③There are TCM syndrome types between 62 and 148,but the sum of frequency of leading syndrome types which includes disease information is mostly no more than 50%.There are TCM syndrome elements between 19 and 47,but the sum of frequency of leading syndrome elements which includes disease information is mostly above 80%.Conclusion: It is convenient to study TCM syndrome from the aspect of TCM syndrome elements,so we should thoroughly keep on studying TCM syndrome elements.
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Systems biology is a new field in biology that aims at system-level understanding of biological systems. Based on the remarkable progress from molecular biology, systems biology focuses its research on investigating the biological systems to form their entirety. Considering the trend of TCM modernization and the coherence on the point of holism between systems biology and TCM, a comprehensive way for the research of TCM modernization was referred, which involves a new concept, componentomics.
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Drug Combinations , Drugs, Chinese Herbal , Medicine, Chinese Traditional , Models, Biological , Neural Networks, Computer , Systems Biology , MethodsABSTRACT
In traditional Chinese medicine,medical practice is combined with qi,which forms special qi-theory.Body fluid which includes blood and Jin-ye is the fluid structure of human being.Nutrient qi and defensive qi depend on these two kinds of body fluid.Defensive qi not only maintains physiological functions of Jin-ye,but also participates pathomechanism such as Jin-ye consumption,retention,etc.The identity of defensive qi and Jin-ye plays an important role in theory and clinic.
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Objective: To investigate the antihypertensive time-effect and dose-effect features of Sancao jiangya decoction(SCD).Methods: The blood pressure of spontaneously hypertensive rats at different time points were measured after treatment with Sancao jiangya decoction of low,middle,high concentration by tailartery blood pressure measurement for conscious rats.Results: The blood pressure was decreased at 2 hours after drug taken,there were significant dose-effect relationship between the antihypertensive effect and the low,middle,high dose.At 4h after drug taken,the high,middle dose had dose-effect correlation,but the low-dose had no antihypertensive effect.Further research on the middledose shows that the blood pressure reduced at 1h after drug taken,and the stable antihypertensive effect was keeping during 1-4h,the blood pressure began to rise at 6h,and got back to the level before drug taken at 8h.Conclusion: To choose the Middle-dose(10.4g crude drug/kg body weight) and 2h after drug taken is appropriate for SCD's use.This result laid a substantial foundation for further research on effects evaluation and mechanism of antihypertensive medicine.