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1.
Neuroscience Bulletin ; (6): 602-616, 2023.
Article in English | WPRIM | ID: wpr-982426

ABSTRACT

Methcathinone (MCAT) belongs to the designer drugs called synthetic cathinones, which are abused worldwide for recreational purposes. It has strong stimulant effects, including enhanced euphoria, sensation, alertness, and empathy. However, little is known about how MCAT modulates neuronal activity in vivo. Here, we evaluated the effect of MCAT on neuronal activity with a series of functional approaches. C-Fos immunostaining showed that MCAT increased the number of activated neurons by 6-fold, especially in sensory and motor cortices, striatum, and midbrain motor nuclei. In vivo single-unit recording and two-photon Ca2+ imaging revealed that a large proportion of neurons increased spiking activity upon MCAT administration. Notably, MCAT induced a strong de-correlation of population activity and increased trial-to-trial reliability, specifically during a natural movie stimulus. It improved the information-processing efficiency by enhancing the single-neuron coding capacity, suggesting a cortical network mechanism of the enhanced perception produced by psychoactive stimulants.


Subject(s)
Mice , Animals , Reproducibility of Results , Neurons , Sensation , Perception
2.
Chinese Journal of Laboratory Medicine ; (12): 1233-1239, 2022.
Article in Chinese | WPRIM | ID: wpr-958649

ABSTRACT

Objective:To establish a candidate reference method for the determination of angiotensin Ⅱ in human plasma by isotope dilution liquid chromatography tandem mass spectrometry (ID-LC-MS/MS) and to evaluate its performance.Methods:Using [ 13C 6- 15N]-angiotensin Ⅱ as the internal standard, the plasma was accurately weighed by gravimetric method and mixed with a certain amount of internal standard. At the same time, enzyme inhibitor was added. After zinc sulfate solution protein precipitation and reversed-phase solid-phase extraction plate treatment, it was analyzed by liquid chromatography tandem mass spectrometry. The multi reaction ion monitoring mode(MRM)was selected by mass spectrometry to detect specific ion fragments of angiotensin Ⅱ and internal standard. The linearity, sensitivity, precision, recovery rate and uncertainty of the performance of the established method were evaluated according to ISO15193. Results:Angiotensin Ⅱ had good linearity in the range of 10-1 000 pg/g ( r=0.999 5), the lower limit of quantification was 7.68 pg/g, the analytical recoveries were 97.14% to 102.85%, intra-batch imprecision≤3.21%, inter-batch imprecision≤2.96%, and total imprecision≤3.67%. Conclusion:A method for the determination of plasma angiotensin Ⅱ was established by ID-LC-MS/MS. The method is accurate and reliable, and is expected to be a reference method for the determination of plasma angiotensin Ⅱ.

3.
Journal of Forensic Medicine ; (6): 423-427, 2019.
Article in English | WPRIM | ID: wpr-985029

ABSTRACT

Objective To analyze a knowledge web of the literature published by Journal of Forensic Medicine from its founding in 1985 to 2018, describe the evolving process of forensic science research and explore the research hotspots and frontiers at present. Methods The literature that was published by Journal of Forensic Medicine from 1985 to 2018 was collected and analyzed in terms of elements, such as emerging research hotspots, high frequency keywords, authors, dispatching units, location of institution and funding, by CiteSpace5.3 information visualization analysis software. Results All disciplines of forensic medicine were continually developing and maturing, and the publication volume of the literature on forensic pathology had the highest weight; in research hotspots, the two categories, research and identification each had their own emphasis; as the main source of contributions to the journal, research institutes accounted for 38.99% of the total number of publications; Shanghai ranked first among all regions with 1 046 articles published. The number of funded articles was generally on the rise, with the number of funded articles published largest in 2015. Conclusion As an authoritative academic journal in the field of forensic science in China, Journal of Forensic Medicine carries the development of forensic science and witnesses the institutional reform of universities and colleges, and offers a wide range of communication and cooperation in terms of technicality and application. Many scholars and scientific research institutions have gained progress continually in various research directions in the form of teamwork; and emerging research hotspots will continue to play a huge role in future practical applications.


Subject(s)
Bibliometrics , China , Forensic Medicine/statistics & numerical data , Forensic Pathology , Forensic Sciences
4.
China Journal of Chinese Materia Medica ; (24): 3862-3866, 2018.
Article in Chinese | WPRIM | ID: wpr-775405

ABSTRACT

Fructus Arctii is a traditional Chinese medicine. The main counterfeit species are the seeds of Arctium tomentosum, Onopordum acanthium, Silybum marianum, Saussurea costus, Amorpha fruticosa. Traditional identification methods or molecular barcoding techniques can identify Fructus Arctii and its counterfeit species. However, the identification of the mixture of it and its spurious species is rarely reported. In this paper, we sequenced the ITS2 sequences of Fructus Arctii and 5 kinds of spurious species mix powder by high-throughput sequencing to identify the mixed powder species and providing new ideas for the identification of Fructus Arctii mix powder. The total DNA in mixed powder was extracted, and the ITS2 sequences in total DNA was amplified. Paired-end sequencing was performed on the DNA fragment of the community using the Illumina MiSeq platform. The sequence was analyzed by the software FLASH, QIIME and GraPhlAn etc. The results showed that the high quality ITS2 sequences of 39910 mix samples were obtained from the mixed samples, of which the total ITS2 sequence of the samples genus was 34 935. Phylogenetic analysis showed that the samples contained Fructus Arctii, A. tomentosum, O. acanthium, S. marianum, S. costus and A. fruticosa. Using ITS2 sequences as DNA barcodes, high-throughput sequencing technology can be used to detect the Fructus Arctii and its spurious specie in mixed powder, which can provide reference for the quality control, safe use of medicinal materials of Fructus Arctii and the identification of mixed powder of traditional Chinese medicine.


Subject(s)
Arctium , Chemistry , Classification , DNA Barcoding, Taxonomic , DNA, Plant , Genetics , DNA, Ribosomal Spacer , Genetics , Drug Contamination , Drugs, Chinese Herbal , Reference Standards , Fabaceae , Fruit , High-Throughput Nucleotide Sequencing , Silybum marianum , Onopordum , Phylogeny , Saussurea
5.
Acta Pharmaceutica Sinica ; (12): 1918-1923, 2018.
Article in Chinese | WPRIM | ID: wpr-780074

ABSTRACT

Traditional Chinese medicine Baitouweng have a long history of application. The pharmacopoeia included dry roots of Pulsatilla chinensis (Bge.) Regel of Ranunculaceae. There are easily confused species in the market circulation, such as P. cernua (Thunb.) Bercht. et Opiz., P. dahurica (Fisch.) Spreng., P. turczaninovii Kryl. et Serg., and P. chinensis (Bge.) Regel var. kissii (Mandl) S. H. Li et Y. H. Huang, etc. In this study, using the method of metagenomics, based on high-throughput sequencing technology, the ITS2 sequence of mixed samples of five species of Baitouweng medicinal materials was sequenced. First, the total DNA extraction of medicinal materials mixing powder, and the ITS2 fragment of total DNA was amplified by PCR. Second, the Illumina MiSeq platform was used to carry out Paired-end sequencing for DNA fragments. Last, using FLASH, QⅡME and GraPhlAn software to arrange and analyze, and clustering analysis with the sequences of uploaded to GenBank by our group in the early stage. The results showed that a total of 53 024 sequences of ITS2 were obtained from the mixed samples, there are 52 295 effective sequences, there are a total of 49 079 of five species of medicinal materials of P. Miller. After the representative sequences and the sequence of uploaded to GenBank by our group in the early stage were clustering analysis, 5 species of Baitouweng medicinal materials were clustered into one branch separately, presenting monophyletic. The results showed that using the high-throughput sequencing technology, using ITS2 sequence as DNA barcode, the mix powder of 5 species of Baitouweng medicinal materials could be effectively identified. It provides a new method and thought for the origin identification of mixed Chinese medicinal materials.

6.
Journal of Medical Postgraduates ; (12): 644-648, 2018.
Article in Chinese | WPRIM | ID: wpr-700889

ABSTRACT

Aptamers,screened by the in vitro process known as Systematic Evolution of Ligands by Exponential Enrichment (SELEX),are single stranded DNA/RNA oligonucleotides that can be folded into three dimensional spatialstructure. They can specif-ically recognize the molecular markers on the cell surface. Compared with monoclonal antibodies,aptamers possess similar or even su-perior characteristics,such as high specificity and affinity,easy for modification,excellet chemical stability,non-toxicity and low im-munogenicity. Due to these advantages,aptamers are emerging as a promising tool to targeted delivery of therapeutic drugs into cells,with low toxicity and high efficacy. They can not only be used as therapeutic agent,specific drug carrier,to deliver such as drugs,nan-oparticles,nucleic acids to targeted cells. This review discusses recent advances of aptamers applied for targeted therapy.

7.
Journal of Forensic Medicine ; (6): 611-616, 2018.
Article in Chinese | WPRIM | ID: wpr-742805

ABSTRACT

Methcathinone, a new cathinone designer drug, which is structurally similar to amphetamine analogs, is a central nervous stimulant.Recently, there has been a worldwide rise in its popularity and abuse, and a growing number of cases with disability or even death is reported in several countries, resulting in public concern.The typical symptoms include accelerated heartbeat, high temperature, anxiety, depression, etc.Forensic studies on its toxicity mechanism are rare.This article reviews its toxicological effects, poisoning symptoms, poisoning and addiction mechanisms, and detection methods, to provide theoretical reference for future studies and guidance for related forensic identification.

8.
Chinese Journal of Clinical and Experimental Pathology ; (12): 1097-1100, 2017.
Article in Chinese | WPRIM | ID: wpr-695025

ABSTRACT

Purpose To investigate the clinicopathologic features and differential diagnosis of oncocytic variant of medullary thyroid carcinoma (MTC).Methods Morphological,immunohistochemical findings on 3 oncocytic variant of MTC cases and electron microscopic findings on 2 of these 3 cases were studied,with review of the relevant literatures.Results The cytoplasm was abundant,eosinophilic and granular with defined margins.The nuclei were enlarged,round to oval.Prominent nucleoli were identified in some cells.Tumor cells formed sheets,trabeculae or follicles with infiltrative pattern.Immunohistochemically,thyroglobulin (TG) was negative in all 3 cases.Calcitonin was positive in 2 cases and negative in 1 case.Ultrastructurally,a large number of mitochondria and various neuroendocrine granules were found in the 2nd and 3rd cases.Conclusion Oncocytic variant of MTC is very rare with variable histopathologic appearances.It should be considered in diagnosing oncocytic lesions of thyroid.A definite diagnosis can be rendered based on comprehensive findings of the immunohistochemistry,serology study and electron microscopy.It needs to avoid misdiagnosis and missed diagnosis.

9.
Chinese Journal of Medical Education Research ; (12): 39-41, 2013.
Article in Chinese | WPRIM | ID: wpr-432506

ABSTRACT

As a rising histological specimen carrier,digital slide has advantages of easy search and fast browse.By digital scanning and stitching of traditional slides and uploading traditional slides as well as pathological information to network servers,the construction of digital slides data can take its own superiority to assist forensic medicine teaching.Combined with our research and teaching experience,this article discussed the application prospects of digital slide technology and digital slides data so as to provide references for the improvement and reformation of forensic medicine education.

10.
Chinese Journal of Cancer ; (12): 620-626, 2011.
Article in English | WPRIM | ID: wpr-294482

ABSTRACT

Matrix metalloproteinase 2 (MMP2) has been shown to play an important role in several steps of cancer development. The -1306C/T polymorphism of the MMP2 gene displays a strikingly lower promoter activity than the T allele, and the CC genotype in the MMP2 promoter has been reported to associate with the development of several cancers. To assess the contribution of the MMP2 -1306C/T polymorphism to the risk of nasopharyngeal carcinoma (NPC), we conducted a case-control study and analyzed MMP2 genotypes in 370 patients with NPC and 390 frequency-matched controls using real-time PCR-based TaqMan allele analysis. We found that subjects with the CC genotype had an increased risk (OR = 1.55, 95% CI = 1.05-2.27) of developing NPC compared to those with the CT or TT genotypes. Furthermore, we found that the risk of NPC was markedly increased in subjects who were smokers (OR = 15.04, 95% CI = 6.65-33.99), heavy smokers who smoked ≥ 20 pack-years (OR = 18.66, 95% CI = 7.67-45.38), or young (<60 years) at diagnosis (OR = 1.52, 95% CI = 1.01-2.29). Our results provide molecular epidemiological evidence that the MMP2 -1306C/T promoter polymorphism is associated with NPC risk, and this association is especially noteworthy in heavy smokers.


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Asian People , Genetics , Carcinoma , Case-Control Studies , China , Epidemiology , Genetic Predisposition to Disease , Genotype , Matrix Metalloproteinase 2 , Genetics , Nasopharyngeal Neoplasms , Epidemiology , Genetics , Pathology , Neoplasm Staging , Polymorphism, Single Nucleotide , Promoter Regions, Genetic , Real-Time Polymerase Chain Reaction , Risk Factors , Smoking
11.
Journal of Forensic Medicine ; (6): 200-204, 2011.
Article in Chinese | WPRIM | ID: wpr-983652

ABSTRACT

OBJECTIVE@#To provide references for forensic expertise by investigating the kinds of toxicant, routes of exposure and manners of poisoning deaths, etc.@*METHODS@#Six hundred and seven autopsy cases of poisoning deaths from 1957 to 2008 in Department of Forensic Medicine, Tongji Medical College (Tongji Forensic Science Identification Center of Hubei), were comparatively reviewed.@*RESULTS@#In 218 cases from 1999 to 2008, more than 50% of decedents were male in the ages of 30-49. The toxicants are usually taken orally and the most common manner of death was accidental. The common substances involved in poisoning death were rodenticide, poisoning gas and insecticide. Compared to the data of 1983-1998 and 1957-1982, the common toxic agents had changed significantly. The number of cases involving insecticide and cyanide poisoning decreased in recent years, and the number of cases of rodenticide, poisoning gas, alcohols poisoning displayed an increase tendency, especially for drugs abuse.@*CONCLUSION@#Poisoning deaths of pesticides remain a major public health problem for a long time and the awareness of prevention need to be raised, especially for the prevention of deaths from multiple poisons.


Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Young Adult , Accidents/statistics & numerical data , Age Distribution , Alcoholic Intoxication/epidemiology , Anesthetics/poisoning , Autopsy , Carbon Monoxide Poisoning/epidemiology , Cause of Death , China/epidemiology , Forensic Medicine , Heroin/poisoning , Hypnotics and Sedatives/poisoning , Pesticides/poisoning , Poisoning/etiology , Retrospective Studies , Rodenticides/poisoning , Sex Distribution , Suicide/statistics & numerical data
12.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 546-50, 2010.
Article in English | WPRIM | ID: wpr-634860

ABSTRACT

The aim of the present study was to assess whether Fourier transform infrared spectrometry (FTIR) micro-spectroscopy could produce distinct spectral information on protein of old myocardial infarction (OMI) and to set them as molecular markers to diagnose atypical OMI. Paraffin-embedded heart samples were derived from victims dying of OMI. In combination with histological stain, FTIR and infrared micro-spectroscopy, the characteristics of OMI were analyzed morphologically and molecularly. The most relevant bands identified were the amide A, B, I and, II showing crucial spectral differences between apparent normal region and OMI region, including the peak position blue shift and the increased intensity of OMI, moreover relative increase in alpha-helix and decrease in beta-sheet of protein secondary structures in OMI. Comparing to single spectral band, the I1650/I1550 ratio was increased and rationally used as a molecular marker for diagnosing OMI. These novel preliminary findings supported further exploration of FTIR molecular profiling in clinical or forensic study, and were in accordance with histopathology.

13.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 235-9, 2010.
Article in English | WPRIM | ID: wpr-634777

ABSTRACT

Corneal opacity is one of the most commonly used parameters for estimating postmortem interval (PMI). This paper proposes a new method to study the relationship between changes of corneal opacity and PMI by processing and analyzing cornea images. Corneal regions were extracted from images of rabbits' eyes and described by color-based and texture-based features, which could represent the changes of cornea at different PMI. A KNN classifier was used to reveal the association of image features and PMI. The result of the classification showed that the new method was reliable and effective.

14.
Journal of Forensic Medicine ; (6): 436-439, 2010.
Article in Chinese | WPRIM | ID: wpr-983608

ABSTRACT

OBJECTIVE@#To develop a rapid and accurate gas chromatography method and investigate the distribution of tramadol in acute poisoned rats for information of samples selection and results evaluation in forensic identification.@*METHODS@#After an oral administration of tramadol at 1140 mg/kg (5 x LD50), concentrations of tramadol in rats' biological fluids and tissues were determined by gas chromatography.@*RESULTS@#The limit of detection of tramadol in blood and urine was 0.1 microg/mL and the limit of detection in liver was 0.1 microg/g. The intra-day precision and inter-day precision were within 3.1% and 5.5% respectively, and the recovery of tramadol in blood was more than 98%. The average levels of tramadol displayed in descending order of heart blood, liver, peripheral blood, urine, vitreous humor, kidney, lung, spleen, heart, brain respectively.@*CONCLUSION@#The established method could meet the requirements for toxicological analysis, and the results of the study suggest that blood, urine, liver, lung and kidney are suitable samples for forensic toxicological analysis in tramadol poisoning cases.


Subject(s)
Animals , Male , Rats , Acute Disease , Administration, Oral , Analgesics, Opioid/urine , Body Fluids/chemistry , Chromatography, Gas/methods , Kidney/metabolism , Liver/metabolism , Random Allocation , Rats, Sprague-Dawley , Reproducibility of Results , Sensitivity and Specificity , Substance Abuse Detection/methods , Tissue Distribution , Tramadol/urine
15.
Journal of Forensic Medicine ; (6): 455-458, 2009.
Article in Chinese | WPRIM | ID: wpr-983526

ABSTRACT

Confocal laser scanning microscopy(CLSM) is a new technique for microscopic imaging, which can collect the transverse section image of the samples and produce three-dimensional reconstruction and present higher spatial resolution than the conventional light microscope. As a precision instrument for the microscopic image, it plays an important role in forensic pathology. The article reviews the recent research achievements from sudden cardiac death, bullet wound and nervous system damage, etc, and explores the potential applications of the forensic pathology research and forensic practice.


Subject(s)
Humans , Calcium/metabolism , Craniocerebral Trauma/pathology , Death, Sudden, Cardiac/pathology , Forensic Pathology/methods , Image Processing, Computer-Assisted , Microscopy, Confocal , Myocardium/ultrastructure , Sensitivity and Specificity , Trauma, Nervous System/pathology , Wounds and Injuries/pathology , Wounds, Gunshot/pathology
16.
Chinese Ophthalmic Research ; (12): 996-999, 2009.
Article in Chinese | WPRIM | ID: wpr-642974

ABSTRACT

Objective Present study aimed to observe the effects of Apoptin gene on killing retinoblastoma HXO-RB_(44) cells and illustrates its mechanisms. Methods Human retinoblastoma cells strain, HXO-RB_(44), was cultured and passaged in RPMI 1640 medium containing bovine serum. Apoptin gene was transfected into HXO-RB_(44) cells by liposome into HXO-RB_(44)/Apoptin, and pcDNA_3 was transfected in HXO-RB_(44)/peDNA_3 group. The expression of Apoptin mRNA was detected using Reverse Transcription Polymerase Chain Reaction (RT-PCR). The expression of protein of Apoptin and p53 were detected by SABC immunohistochemistry. The growth rate of HXO-RB_(44) cells was studied by constructing the growth curve and calculated as the formula: inhibitory rate = 1-cell number in experiment group/cell number in control group x 100%. Cellular apoptosis was determined by flow cytometry. Results The RT-PCR result showed the 450 kb specific band in UXO-RB_(44)/Apoptin group and absent amplification result in HXO-RB_(44) group and HXO-RB_(44)/pcDNA_3 group. The difference in SABC-positive cell number between HXO-RB_(44)/Apoptin group and control group was statistically significant (P < 0. 05). The growth of HXO-RB_(44) cells was significantly inhibited in HXO-RB_(44)/Apoptin group compared with control group (P < 0.05). Apoptosis cells increased significantly. The apoptosis rate was 38. 5% . Conclusion Apoptin gene could inhibit the growth of HXO-RB_(44) cells effectively. Up-regulation of expression of p53 gene might not be one of cell apoptosis mechanisms.

17.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 499-503, 2008.
Article in English | WPRIM | ID: wpr-634933

ABSTRACT

In order to investigate the effects of aconitine on [Ca2+] oscillation patterns in cultured myocytes of neonatal rats, fluorescent Ca2+ indicator Fluo-4 NW and laser scanning confocal microscope (LSCM) were used to detect the real-time changes of [Ca2+] oscillation patterns in the cultured myocytes before and after aconitine (1.0 micromol/L) incubation or antiarrhythmic peptide (AAP) and aconitine co-incubation. The results showed under control conditions, [Ca2+] oscillations were irregular but relatively stable, occasionally accompanied by small calcium sparks. After incubation of the cultures with aconitine, high frequency [Ca2+] oscillations emerged in both nuclear and cytoplasmic regions, whereas typical calcium sparks disappeared and the average [Ca2+] in the cytoplasm of the cardiomyocyte did not change significantly. In AAP-treated cultures, intracellular [Ca2+] oscillation also changed, with periodic frequency, increased amplitudes and prolonged duration of calcium sparks. These patterns were not altered significantly by subsequent aconitine incubation. The basal value of [Ca2+] in nuclear region was higher than that in the cytoplasmic region. In the presence or absence of drugs, the [Ca2+] oscillated synchronously in both the nuclear and cytoplasmic regions of the same cardiomyocyte. It was concluded that although oscillating strenuously at high frequency, the average [Ca2+] in the cytoplasm of cardiomyocyte did not change significantly after aconitine incubation, compared to the controls. The observations indicate that aconitine induces the changes in [Ca2+] oscillation frequency other than the Ca2+ overload.


Subject(s)
Aconitine/pharmacology , Animals, Newborn , Calcium Signaling/drug effects , Cells, Cultured , Myocytes, Cardiac/cytology , Myocytes, Cardiac/metabolism , Rats, Sprague-Dawley
18.
Chinese Journal of Surgery ; (12): 1037-1040, 2007.
Article in Chinese | WPRIM | ID: wpr-340866

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the effects of surgical treatment for gastrointestinal stromal tumors (GISTs) and influential factors of survival.</p><p><b>METHODS</b>The clinical data and the tissue slices including immunohistochemical staining of 153 cases of GISTs from January 1990 to March 2006 were rechecked retrospectively. All patients were followed up carefully. More attention was paid to the surgical effects and the influential factors of survival.</p><p><b>RESULTS</b>The overall survival rates at 1-, 2-, 3-, 4- and 5-year were 94.9%, 83.3%, 73.3%, 70.5% and 64.3%, respectively. The median survival time for patients with tumor resected completely was 66.0 months, and the 2- and 5-year survival rate were 89.4% and 70.9% respectively. The median survival time was 23.8 months for the patients with tumor resected partly, and only two of these patients survived over 2 years. Gender, tumor sites, preoperative metastasis, tumor size, pathological type, karyokinesis and recurrence and metastasis were related with survival rates for the patients with tumor resected completely on univariate analysis, but tumor size, pathology type, recurrence and metastasis were related with survival rates on Cox regression multivariate analysis (P < 0.05).</p><p><b>CONCLUSIONS</b>Surgery should still be the main therapy for GISTs. Local complete resection is the principal treatment. The survival cannot be improved by extensive resection and lymph nodes clearance.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Antigens, CD34 , Follow-Up Studies , Gastrointestinal Stromal Tumors , Metabolism , Mortality , General Surgery , Immunohistochemistry , Kaplan-Meier Estimate , Proto-Oncogene Proteins c-kit , Retrospective Studies , Survival Rate , Treatment Outcome
19.
Biomedical and Environmental Sciences ; (12): 87-95, 2006.
Article in English | WPRIM | ID: wpr-229720

ABSTRACT

<p><b>OBJECTIVE</b>To develop a safe and novel immunoadjuvant to enhance the immunity and resistance of animals against E. coli infection.</p><p><b>METHODS</b>An 88-base immunostimulatory oligodeoxynuleotide containing eleven CpG motifs (CpG ODN) was synthesized and amplified by PCR. The chitosan nanoparticle (CNP) was prepared by ion linking method to entrap the CpG ODN that significantly promotes the proliferation of lymphocytes of pig in vitro. Then the CpG-CNP was inoculated into 21-day old Kunming mice, which were orally challenged with virulent K88/K99 E. Coli 35 days after inoculation. Blood was collected from the tail vein of mice on days 0, 7, 14, 21, 28, 35, 42, and 49 after inoculation to detect the changes and content of immunoglobulins, cytokines and immune cells by ELISA, such as IgG, IgA, IgM, IL-2, IL-4, and IL-6.</p><p><b>RESULTS</b>The CpG provoked remarkable proliferation of lymphocytes of pig in vitro in comparison with that of control group (P < 0.05). The inoculation with CpG-CNP significantly raised the content of IgG, IgM, and IgA in the sera of immunized mice (P < 0.05). The levels of IL-2, IL-4, and IL-6 in the mice significantly increased in comparison with those in controls (P < 0.05), so was the number of white blood cells and lymphocytes in immunized mice. The humoral and cellular immunities were significantly enhanced in immunized mice, which resisted the infection of E. coli and survived, while the control mice manifested evident symptoms and lesions of infection.</p><p><b>CONCLUSIONS</b>CpG-CNP can significantly promote cellular and humoral immunity and resistance of mice against E. coil infection, and can be utilized as an effective adjuvant to improve the immunoprotection and resistance of porcine against infectious disease.</p>


Subject(s)
Animals , Female , Mice , Adjuvants, Immunologic , Antibodies, Bacterial , Blood , Biocompatible Materials , Chitosan , CpG Islands , Escherichia coli , Virulence , Escherichia coli Infections , Allergy and Immunology , Microbiology , Immunity, Cellular , Interleukins , Lymphocyte Activation , Nanoparticles , Oligodeoxyribonucleotides , Swine , Vaccination
20.
Biomedical and Environmental Sciences ; (12): 315-322, 2006.
Article in English | WPRIM | ID: wpr-229682

ABSTRACT

<p><b>OBJECTIVE</b>To study the regulating effects of a novel CpG oligodeoxynuleotide and the synergistic effect of chitosan-nanoparticles (CNP) with CpG on immune responses of mice, which were used to develop a novel immunoadjuvant to boost immune response to conventional vaccines.</p><p><b>METHODS</b>A novel CpG ODN containing 11 CpG motifs was synthesized and its bioactivities to stimulate the proliferation of lymphocytes of pig in vitro were detected. Then it was entrapped with CNP prepared in our laboratory by the method of ionic cross linkage, and immunized Kunming mice were co-inoculated with paratyphoid vaccine. The peripheral blood was collected weekly from the tail vein of inoculated mice to detect the contents of IgG, IgA, IgM, and specific antibody against salmonella as well as the levels of interleukin-2 (IL2), IL-4, and IL-6 by SABC-ELISA assay. The numbers of leucocytes, monocytes, granuloytes, and lymphocytes were calculated separately using the routine method. The experimental mice were orally challenged with virulent salmonella 35 days after inoculation.</p><p><b>RESULTS</b>This CpG ODN could remarkably provoke the proliferation of lymphocytes of pig in vitro in contrast with the control (P < 0.05). Compared with those of the control, immunoglobulins, including IgG, IgA, IgM, and specific antibodies to paratyphoid vaccine, increased significantly in sera from the CpG or CpG-CNP-vaccinated mice (P < 0.05). IL-2, IL-4, and IL-6 increased remarkably in sera from immunized mice (P < 0.05). The leucocytes, monocytes, granuloytes, and lymphocytes of the mice immunized with CpG or CpG-CNP were also increased in number (P < 0.05). After the challenge, these immunity values were elevated in the mice vaccinated with CpG or CpG-CNP. The immunized mice all survived, while the control mice fell ill with evident lesions with diffuse hemorrhage in stomach, small intestine, and peritoneum.</p><p><b>CONCLUSIONS</b>CpG ODN entrapped with CNP is a promising effective immunoadjuvant for vaccination, which promotes humoral and cellular immune responses, enhances immunity and resistance against salmonella by co-administration with paratyphoid vaccine.</p>


Subject(s)
Animals , Mice , Adjuvants, Immunologic , Pharmacology , Antibodies, Bacterial , Blood , Cell Proliferation , Chitosan , Chemistry , Drug Therapy, Combination , Enzyme-Linked Immunosorbent Assay , Immunoglobulin Isotypes , Blood , Interleukins , Blood , Lymphocyte Activation , Lymphocytes , Cell Biology , Nanoparticles , Chemistry , Oligodeoxyribonucleotides , Pharmacology , Paratyphoid Fever , Allergy and Immunology , Salmonella , Physiology , Salmonella Infections, Animal , Allergy and Immunology , Swine , Allergy and Immunology , Typhoid-Paratyphoid Vaccines , Allergy and Immunology
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