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1.
Chinese Pharmacological Bulletin ; (12): 1566-1576, 2023.
Article in Chinese | WPRIM | ID: wpr-1013745

ABSTRACT

Aim To investigate the mechanism of corn silk decoction on diabetic nephropathy (DN) rats using metabolomics technology. Methods DN rat model was established by feeding with high-sugar and high-fat diet, combined with intraperitoneal injection of low dose streptozotocin. Renal organ index, fasting blood glucose, albumin creatinine ratio, serum creatinine, blood urea nitrogen, triglyceride, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol indexes were measured, and the pathological changes of renal tissues were also observed to evaluate the intervention effect of corn silk on DN model rats. Further, UPLC/Q-TOF-MS technology was used to screen potential biomarkers in renal tissues and urine, combined with principal component analysis (PC A) and orthogonal partial least squares discriminant analysis (OPLS-DA). After identification by HM-DB and KEGG database, the biomarkers were imported into MetaboAnalyst for metabolic pathway analysis. Results All indexes and pathological damage of kidneys were improved in groups with different doses of corn silk, indicating that corn silk had a good intervention effect on DN. Metabolomic analysis showed that 18 biomarkers could be significantly called back by corn silk, and it involved 18 metabolic pathways mainly including phenylalanine, tyrosine and tryptophan biosynthesis, riboflavin metabolism, arachidonic acid metabolism, and tyrosine metabolism. Conclusions The mechanism of corn silk decoction intervention on DN may be related to amino acid metabolism, riboflavin metabolism, and arachidonic acid metabolism.

2.
China Journal of Chinese Materia Medica ; (24): 1417-1429, 2021.
Article in Chinese | WPRIM | ID: wpr-879047

ABSTRACT

In this experiment, ultra high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to analyze and identify chemical constituents of Ginseng-Douchi(GD) compound fermentation, and explore the conversion rules of ginsenosides and soybean isoflavones after compound fermentation. Waters Acquity UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) was adopted, with 0.1% formic acid aqueous solution(A)-0.1% formic acid acetonitrile solution(B) as mobile phase for gradient elution; electrospray ion source(ESI) was used to collect data in positive and negative ion modes; according to the exact mass number, the secondary spectrum comparison of the database and the existing literature reports, Peakview 2.0/masterview 1.0 software was used to determine the common ion structure formula. Finally, a total of 133 chemical constituents were analyzed and identified from the GD. Ginseng saponins and isoflavone glycosides were significantly converted after fermentation. Among them, peak areas of prototype ginsenosides Rk_3, Rh_1, Rh_2, Rh_3, daidzin, glycitin and genistin decreased significantly; whereas peak areas of se-condary ginsenoside Rb_1, Rb_2, Rk_1, glycitein, genistein and daidzein increased significantly. In this experiment, liquid-mass spectrometry technique was used to investigate the conversion of active ingredients of GD compound fermented products after co-fermentation, so as to provide a scientific basis for elucidating pharmacodynamics material basis and quality control.


Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Fermentation , Panax , Tandem Mass Spectrometry
3.
China Journal of Orthopaedics and Traumatology ; (12): 867-872, 2020.
Article in Chinese | WPRIM | ID: wpr-827241

ABSTRACT

OBJECTIVE@#To explore the biomechanical characteristics of "three-dimensional balanced manipulation" for the treatment of cervical spondylotic radiculopathy(CSR).@*METHODS@#A CSR patient was treated with "three-dimensional balanced manipulation", and the mechanical changes during the manipulation were monitored by mechanical testing system. Using spiral CT to scan the neck of the patient to obtain DICOM data. The three-dimensional finite element model of cervical spondylotic radiculopathy was established by using Mimics software, Geographic Studio software. The "three-dimensional balance manipulation" was simulated and loaded, and the mechanical parameters of each part were replaced into the finite element model, and the finite element analysis was carried out by using ANSYS software to study the internal stress changes and displacement deformation of vertebral body and intervertebral disc under the action of "three-dimensional balance manipulation".@*RESULTS@#The established C-C finite element model of the CSR patient consisted of 5 vertebrae, 4 intervertebral discs and 3 ligaments, involving 153 471 nodes and 64 978 units. The stress of C-C vertebral body was mainly located in anterior and root of C spinous processes, arch, vertebral arch and the combination of the two after full loading of manipulation, and the maximum stress was 17.781 MPa. The deformation sites were mainly concentrated in articular processes and anterior transverse processes of C, superior articular processes and transverse processes of C, articular processes of C. The stress of C-C intervertebral disc mainly distributed in the anterior part of C intervertebral disc and the nucleus pulposus of C and C. The displace mentextended to the middle and posterior part of C nucleus pulposus, around the nucleus of C and C and anterior part of cervical intervertebral disc.@*CONCLUSION@#The establishment of three-dimensional finite element model of C-C cervical spondylotic radiculopathy can simulate the geometry and material properties of cervical spine, and also accurately reflects the biomechanical characteristics of cervical spine, verifys the internal mechanism of "three-dimensional balanced manipulation" on CSR, proves the safety and effectiveness of treatment, guides more standardized manipulation, and avoids medical accidents.


Subject(s)
Humans , Biomechanical Phenomena , Cervical Vertebrae , Finite Element Analysis , Intervertebral Disc , Radiculopathy , Range of Motion, Articular , Spondylosis
4.
Chinese Traditional and Herbal Drugs ; (24): 3774-3783, 2019.
Article in Chinese | WPRIM | ID: wpr-850908

ABSTRACT

Objective: To identify the common compounds and variance compounds of Sophorae Fructus from different regions by ultra performance liquid chromatography coupled with time-of-fight mass spectrometry (UFLC-Q-TOF-MS). Methods: The separation was performed on Waters Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm), with a mobile phase using water with 0.1% formic acid (A) and acetonitrile with 0.1% formic acid (B) for gradient elution; Q-TOF/MS and electrospray ion (ESI) source were applied for the analysis under the positive ion mode and the negative ion mode; 1 000 ions were extracted through Markerview 1.2.1 software from Sophorae Fructus of 11 different regions. And common ions (compounds) were selected according to the following principles: One ion can be detected in all samples, and most of the peak area was greater than the 1 × 104; The variance ions (compounds) were selected according to principal components analysis. The formula of common ions was then determined by accurate mass and isotopic abundance ratio from target screening function of Peakview 2.0/masterview 1.0 software, and its structure were determined by analysis of MS/MS fragment or comparison with standard substances and references. Results: A total of 24 common compounds and 21 variance compounds were identified and inferred in Sophorae Fructus from different producing areas. Conclusion: UPLC-Q-TOF/MS method which develops a new strategy can identify the main chemical constituents from Sophorae Fructus rapidly and accurately. The determination of the main common components lays a foundation for the selection of quality evaluation indexes and the in-depth study of pharmacodynamic substances. The variance components can also be used as one of the bases for the identification of origin and authentic evaluation of Sophorae Fructus.

5.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 525-545, 2018.
Article in English | WPRIM | ID: wpr-773587

ABSTRACT

The present study was designed to identify and characterize the major constituents in Juglans mandshurica Maxim. A simple, efficient and sensitive ultra performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-ESI-Q-TOF/MS) method was established and validated under positive and negative ion modes. The separation was performed on a Waters ACQUITY UPLC BEH C column (2.1 mm × 100 mm, 1.7 μm) by gradient elution with a mobile phase (Phase A: 0.1% aqueous formic acid solution, Phase B: 0.1% formic acid acetonitrile solution). A total of 165 compounds were rapidly selected by Targeted and Non-Targeted Peak Finding approaches, and then tentatively identifled by comparing with reference substances or inferred through mass spectrometry fragment ion analysis and literature data. These compounds included 68 naphthalenequinones, 20 diarylheptanoids, 29 flavonoids, 20 triterpenes, and 28 phenolic acids. In conclusion, the present study provided an effective approach to identifying components in complex matrices of herbal medicines such as Juglans mandshurica Maxim.


Subject(s)
Chromatography, High Pressure Liquid , Databases, Factual , Diarylheptanoids , Chemistry , Drugs, Chinese Herbal , Chemistry , Flavonoids , Chemistry , Fruit , Chemistry , Hydroxybenzoates , Chemistry , Juglans , Chemistry , Molecular Structure , Naphthoquinones , Chemistry , Reproducibility of Results , Tandem Mass Spectrometry , Triterpenes , Chemistry
6.
Chinese Medical Equipment Journal ; (6): 46-48, 2018.
Article in Chinese | WPRIM | ID: wpr-699963

ABSTRACT

Objective To develop a pottery frame to facilitate warm acupuncture moxibustion.Methods The frame was made of clay,and was composed of a moxa wool retaining table,a handle,a base and a support.The moxa wool holding table had a hollow structure wide at the top and narrow at the bottom,and there was a slot at its middle to hold the needle;while the base had a hollow structre narrow at the top and wide at the bottom,and there were several air vents at its inner wall to accelerate the full combustion of the moxa wool.Results The frame avoided patient burn due to dropped spark or overweight moxa cone during traditional warm acupuncture moxibustion therapy, and relieved the patients' pain and medical staffs' workload. Conclusion The frame has simple structure, easy operation, high safety, high patient satisfaction and high practicability.

7.
Journal of Southern Medical University ; (12): 174-180, 2018.
Article in Chinese | WPRIM | ID: wpr-299282

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the clinical effect of spinal cord decompression and lavage therapy on chronic cervical spinal cord injury and explore the possible mechanism.</p><p><b>METHODS</b>Fifty-seven patients with chronic cervical spinal cord injury treated in our hospital from January, 2008 to January, 2015 were enrolled, including 17 with multilevel cervical disc herniation, 25 with long segmental ossification of the posterior longitudinal ligament, 13 with hypertrophy or calcification of neck ligamentum flavum, and 2 with old cervical fractures. Open-door spinal canal laminoplasty via a posterior approach and decompression in simple extramedullary decompression was performed in 31 cases (group A), and open-door spinal cord incision decompression via a posterior approach, saline irrigation, and spinal canal laminoplasty in intramedullary decompression was performed in 26 cases (group B). The pre-operative cerebrospinal fluid in group B patients was collected to examine the inflammatory factors. All the patients were followed up and evaluated for pre- and postoperative JOA scores to calculate the improvement rate with regular examinations by X-ray, CT or MRI.</p><p><b>RESULTS</b>Imaging examinations 2 weeks after the operation showed obvious relief of the primary lesion in both groups, and the improvement of high signals was better in group B than in group A. The mean improvement rate at 12 months after the operation was 52.33% in group A and 61.52% in group B (P<0.05), and the mean JOA score was significantly higher in group B than in group A (14.80∓1.51 vs 13.58∓0.56; P<0.05). Cerebrospinal fluid leakage occurred in 3 cases, epidural hematoma in 2 cases, internal fixation loosening in 1 case in group A; portal shaft fracture and internal fixation loosening occurred in 1 case in group B. Postoperative recovery time was shorter in group B and entered the platform phase in 3 months. The inflammatory factors IFN-γ, IL-17F, IL-6 and sCD40L were all significantly higher than the normal levels after spinal cord injury, and the increment of IL-6 was the most conspicuous (P<0.05).</p><p><b>CONCLUSION</b>Intramedullary and extramedullary decompression can achieve better outcomes than extramedullary decompression in patients with chronic cervical cord injury. This may be related not only to relieving adhesions and secondary compression by cutting the dura under the microscope, but also to removal of local inflammatory factors.</p>

8.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 525-545, 2018.
Article in English | WPRIM | ID: wpr-812376

ABSTRACT

The present study was designed to identify and characterize the major constituents in Juglans mandshurica Maxim. A simple, efficient and sensitive ultra performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-ESI-Q-TOF/MS) method was established and validated under positive and negative ion modes. The separation was performed on a Waters ACQUITY UPLC BEH C column (2.1 mm × 100 mm, 1.7 μm) by gradient elution with a mobile phase (Phase A: 0.1% aqueous formic acid solution, Phase B: 0.1% formic acid acetonitrile solution). A total of 165 compounds were rapidly selected by Targeted and Non-Targeted Peak Finding approaches, and then tentatively identifled by comparing with reference substances or inferred through mass spectrometry fragment ion analysis and literature data. These compounds included 68 naphthalenequinones, 20 diarylheptanoids, 29 flavonoids, 20 triterpenes, and 28 phenolic acids. In conclusion, the present study provided an effective approach to identifying components in complex matrices of herbal medicines such as Juglans mandshurica Maxim.


Subject(s)
Chromatography, High Pressure Liquid , Databases, Factual , Diarylheptanoids , Chemistry , Drugs, Chinese Herbal , Chemistry , Flavonoids , Chemistry , Fruit , Chemistry , Hydroxybenzoates , Chemistry , Juglans , Chemistry , Molecular Structure , Naphthoquinones , Chemistry , Reproducibility of Results , Tandem Mass Spectrometry , Triterpenes , Chemistry
9.
Chinese Traditional and Herbal Drugs ; (24): 657-667, 2017.
Article in Chinese | WPRIM | ID: wpr-852968

ABSTRACT

Objective: To identify the major chemical constituents in Cortex Juglandis Mandshuricae by ultra performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Methods: The separation was performed on Waters Acquity UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm), with a mobile phase using water with 0.1% formic acid (A) and acetonitrile with 0.1% formic acid (B) for gradient elution; Q-TOF/MS and electrospray ion (ESI) source was applied for the analysis under the positive and negative ion modes; Target search and non-target search were performed by Peakview 2.0/masterview1.0 or Markerview 1.2.1 software. Then the formulas of compounds were determined by accurate mass and isotopic abundance ratio from target screening function of software. Their structures were determined by analysis of MS/MS fragment or comparison with standard substances and references. Results: Forty-four major chemical compounds including 13 naphthalene quinones, 3 diarylheptanoids, 15 flavonoids, and 13 other compounds were identified or inferred in Cortex Juglandis Mandshuricae. Conclusion: UPLC-Q-TOF/MS method which develops a new strategy can identify the main chemical constituents from Cortex Juglandis Mandshuricae rapidly and accurately, main chemical constituents can be used for quality evaluation and pharmacological research.

10.
China Journal of Chinese Materia Medica ; (24): 3112-3120, 2017.
Article in Chinese | WPRIM | ID: wpr-335886

ABSTRACT

To analyze the dynamic changes in components in exocarp of Juglans mandshurica at different browning periods. Twenty-six batches of exocarp of J. mandshurica samples from thirteen browning periods were assessed by UPLC-Q-TOF-MS/MS. The formula of different compounds were determined by accurate mass and isotopic abundance ratio from target screening function of Peakview 2.0/masterview1.0 software. Then their structures were determined by analysis of MS/MS fragment or comparison with standard substances and references. The contents of chemical components were changed significantly in different browning periods and twenty five compounds were identified or inferred. Of the 13 naphthoquinone compounds, the contents of 6 compounds with similar parent nucleus as juglone and 3 naphthoquinone glycosides compounds were decreased significantly, and 4 naphthoquinone derivatives such as regiolone were produced; the contents of four flavones and two phenolic acids compounds were decreased significantly; and the contents of 6 diarylheptanoids compounds were increased significantly. UPLC-Q-TOF/MS method can be used to identify and analyze the chemical constituents from exocarp of J. mandshurica rapidly and accurately, and analyze the rules of dynamic changes, to reveal the browning of Chinese medicinal materials and its effects on compositions of fruits and vegetables.

11.
China Journal of Chinese Materia Medica ; (24): 3379-3388, 2016.
Article in Chinese | WPRIM | ID: wpr-307148

ABSTRACT

The changes in effective components of Juglans mandshurica at different harvest periods were analyzed by UPLC-Q-TOF-MS/MS. Eighteen batch samples of J. mandshurica from six harvest periods were assessed by multivariate statistical analysis with Markerview software. The formula of different compounds were determined by accurate mass and isotopic abundance ratio from target screening function of Peakview 2.0/Masterview1.0 software. Then their structure were determined by analysis of MS/MS fragment or comparison with standard substances and references. Naphthoquinone are the major markers in samples of Juglans mandshurica from different harvest periods. Thirty-eight of naphthalenequinones were identified or inferred in J. mandshurica and contents decline gradually. UPLC-Q-TOF-MS method which develops a new strategy can identify and analyze chemical constituents from J. mandshurica rapidly and accurately, main chemical constituents can be used for quality evaluation and efficacy material research. The dynamic changes in the metabolite accumulation of J. mandshurica the basic data for harvesting medicinal plants at different times.

12.
Journal of Southern Medical University ; (12): 321-326, 2016.
Article in Chinese | WPRIM | ID: wpr-264047

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of miR-181b on the migration and invasion of osteosarcoma cells.</p><p><b>METHODS</b>Three cultured osteosarcoma cell lines and MG-63 cells transfected with miR-181b inhibitor were examined for miR-181b expression using qRT-PCR analysis. The cell migration and invasion of the transfected cells were assessed with Transwell assay. The targets of miR-181b were predicted using a miRNA target prediction software and the results were verified with luciferase reporter assay. The target protein expression in osteosarcoma cells lines was determined by Western blotting, and the cell migration and invasion changes following inhibition of miR-181b or its target protein were assessed using Transwell assay.</p><p><b>RESULTS</b>All the 3 osteosarcoma cells lines showed significantly up-regulated miR-181b expression. Inhibition of miR-181b expression obviously suppressed the migration and invasion of MG-63 cells. Based on luciferase reporter assay, N-myc downstream regulated gene 2 (NDRG2) was identified as the direct target gene of miR-181b, and inhibition of NDRG2 expression significantly reversed the effect of miR-181b on cell migration and invasion in MG-63 cells.</p><p><b>CONCLUSION</b>miR-181b is over-expressed in osteosarcoma cells, and inhibition of miR-181b, which directly targets NDRG2, can suppress the migration and invasion of osteosarcoma cells.</p>


Subject(s)
Humans , Bone Neoplasms , Genetics , Pathology , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , MicroRNAs , Genetics , Metabolism , Neoplasm Invasiveness , Osteosarcoma , Genetics , Pathology , Tumor Suppressor Proteins , Genetics , Metabolism
13.
Biomedical and Environmental Sciences ; (12): 1-12, 2015.
Article in English | WPRIM | ID: wpr-264625

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of electronspun PLGA/HAp/Zein scaffolds on the repair of cartilage defects.</p><p><b>METHODS</b>The PLGA/HAp/Zein composite scaffolds were fabricated by electrospinning method. The physiochemical properties and biocompatibility of the scaffolds were separately characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), and fourier transform infrared spectroscopy (FTIR), human umbilical cord mesenchymal stem cells (hUC-MSCs) culture and animal experiments.</p><p><b>RESULTS</b>The prepared PLGA/HAp/Zein scaffolds showed fibrous structure with homogenous distribution. hUC-MSCs could attach to and grow well on PLGA/HAp/Zein scaffolds, and there was no significant difference between cell proliferation on scaffolds and that without scaffolds (P>0.05). The PLGA/HAp/Zein scaffolds possessed excellent ability to promote in vivo cartilage formation. Moreover, there was a large amount of immature chondrocytes and matrix with cartilage lacuna on PLGA/HAp/Zein scaffolds.</p><p><b>CONCLUSION</b>The data suggest that the PLGA/HAp/Zein scaffolds possess good biocompatibility, which are anticipated to be potentially applied in cartilage tissue engineering and reconstruction.</p>


Subject(s)
Animals , Female , Humans , Male , Young Adult , Biocompatible Materials , Bone Development , Physiology , Cartilage , Cells, Cultured , Durapatite , Chemistry , Lactic Acid , Chemistry , Mesenchymal Stem Cells , Physiology , Polyglycolic Acid , Chemistry , Regeneration , Physiology , Tissue Scaffolds , Chemistry , Zein , Chemistry
14.
Chinese Journal of Medical Genetics ; (6): 29-32, 2011.
Article in Chinese | WPRIM | ID: wpr-234324

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the RHD zygosity of Rh(D)-positive Chinese Hans in order to study the mother-fetus Rh isoimmunization prophylaxis.</p><p><b>METHODS</b>Rh(D) blood group of 31 115 donors were serotyped, and the RHD zygosities were analyzed, or determined through a PCR method for 3628 donors of Rh(D)-positive individuals.</p><p><b>RESULTS</b>Among the 31 115 donors, 99 were tested Rh(D)-negative by indirect antiglobulin test (IAT) (0.318%). The d frequency was 0.056 41, D was 0.943 59, and Dd heterozygosity was 0.106 45 (10.6%). However the rate was 0.090 32 (about 9.0%) after excluding DEL (IAT-negative). For the 3628 PCR tested donors, 3383 were DD (93.2%), 245 were Dd (6.8%). After excluding nonfunctional RHD alleles, 7.4% of the donors were carrying one functional RHD. It showed that an Rh(D)-negative Chinese Han woman gives an Rh(D)-negative child at a rate of 3.7%-4.5% when her husband is Rh(D)-positive.</p><p><b>CONCLUSION</b>Fetus Rh(D) genotyping may be unnecessary for Chinese Hans if invasive operation was needed for prenatal diagnosis. The Rh prophylaxis could be chosen assuming an Rh(D)-positive fetus.</p>


Subject(s)
Adolescent , Adult , Female , Humans , Infant, Newborn , Male , Middle Aged , Asian People , Genetics , China , Ethnology , Polymerase Chain Reaction , Rh-Hr Blood-Group System , Blood , Genetics
15.
Chinese Medical Journal ; (24): 188-192, 2009.
Article in English | WPRIM | ID: wpr-311893

ABSTRACT

<p><b>BACKGROUND</b>Celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, is a non-steroidal anti-inflammatory drug used as an adjuvant to sensitize cancer cells to apoptosis. However, in rats suffering from acute rejection, celecoxib reduced apoptosis of myocardial cells. We hypothesize that celecoxib reduces myocardial apoptosis either by inducing apoptosis in peripheral blood lymphocytes (PBLs) or by altering the percentage of CD4(+) and CD8(+) lymphocytes.</p><p><b>METHODS</b>After cardiac transplantation, rats were administered intragastrically with celecoxib (50 mg/kg per day) for 3, 5 or 7 days, at which time the graft was excised and evaluated for organ rejection. In addition, PBLs were isolated from the blood to determine PBLs apoptosis, and the percentage of CD4(+) and CD8(+) lymphocytes.</p><p><b>RESULTS</b>Celecoxib induced PBLs apoptosis in 3 days, but protected the cells from apoptosis at 5 and 7 days. Also, the percentage of CD4(+) lymphocytes decreased only at 3 days, but a reduction in the percentage of CD8(+) lymphocytes was not seen until 7 days after the transplant surgery. Celecoxib only decreased acute rejection at 5 days, with no discernible difference in rejection after 3 and 7 days.</p><p><b>CONCLUSIONS</b>The results suggested that celecoxib displayed a multiple physiological function in a time-dependent manner.</p>


Subject(s)
Animals , Male , Rats , Anti-Inflammatory Agents , Pharmacology , Apoptosis , Celecoxib , Cells, Cultured , Graft Rejection , Allergy and Immunology , Heart Transplantation , Allergy and Immunology , Lymphocytes , Cell Biology , Allergy and Immunology , Membrane Potential, Mitochondrial , Pyrazoles , Pharmacology , Rats, Sprague-Dawley , Rats, Wistar , Sulfonamides , Pharmacology , Transplantation, Homologous , Allergy and Immunology
16.
Journal of Experimental Hematology ; (6): 1024-1028, 2006.
Article in Chinese | WPRIM | ID: wpr-282739

ABSTRACT

This study was aimed to investigate the molecular genetic basis and serological phenotype of Rh weak D type 15 individuals. Samples were identified by serological tests and genotyped by sequence specific primer-PCR (SSP-PCR), and were sequenced to detect the changes of all ten RHD exons. The number of gene RHD was detected through SSP-PCR. The results showed that in tested individuals of weak D type confirmed by the IAT, 18 cases (56% in weak D) were weak D type 15. Rh factors found in 2 weak D type 15 individuals (11%) were C+c+E+e; Rh factors found in 2 weak D type 15 individuals (11%) were C+c+E-e+; others (78%) were c-c+E+e+. The results by serological tests were consistent with the results genotyped by PCR-SSP method. In all 18 samples, the sequencing result revealed a gene mutation 845G > A at the exon 6 of the RHD and the point mutation changed amino acid G282D of the RhD polypeptide. The zygosity test demonstrated that 2 out of 18 weak D type 15 individuals were RHD(+)/RHD(+) homozygous (two DCe/DcE), 16 cases were RHD(+)/RHD(-) heterozygous (two DCe/dce and fourteen DcE/dce). It is concluded that Weak D type 15 is predominant in weak D individuals of Chinese Han Nationality, and most of them are heterozygous with various RH haplotypes.


Subject(s)
Humans , Asian People , Genetics , Base Sequence , Blood Donors , China , Ethnology , Erythrocytes , Allergy and Immunology , Exons , Genetics , Genotype , Haplotypes , Molecular Sequence Data , Phenotype , Point Mutation , Polymerase Chain Reaction , Methods , Polymorphism, Genetic , Rh-Hr Blood-Group System , Genetics , Allergy and Immunology , Sequence Analysis, DNA
17.
Journal of Experimental Hematology ; (6): 889-895, 2005.
Article in Chinese | WPRIM | ID: wpr-343864

ABSTRACT

To study the genetic polymorphism of HPA 1-16 platelet antigen alleles among unrelated volunteer donors and establish a typed platelet donor panel in Handan, typing was perfomed by polymerase chain reaction using sequence-specific primers (SSP-PCR); 148 random unrelated blood donors in Handan were genotyped for each of the HPA 1-16 antigen. The gene frequencies were analyzed and the genetype frequencies were determined by direct counting, and these data were compared with HPA distribution among various population by the chi-square test. The results indicated that HPA-1a, 2a, 4a-14a, 16a genes were found among the 16 HPAs in every sample tested. Monomorphic HPA-4a, 7a-14a, 16a were found in the samples. For HPA-1, 2, 5 and 6, a/a homozygosity was predominant with frequencies of 0.9595, 0.8108, 0.9865, 0.9797, respectively, and none of HPA b/b was found in the samples. HPA-1b, 2b, 5b, 6b were rarely found among subjects. HPA-15 had the greatest heterozygosity with a gene frequency of 0.2230, 0.5270, 0.2500 for HPA15a/15a, HPA15a/15b, HPA15b/15b, respectively. HPA-3 showed the second greatest heterozygosity with a gene frequency of 0.3851, 0.5135, 0.1014 for HPA3a/3a, HPA3a/3b, HPA3b/3b, respectively. HPA genotype frequencies showed a good fit to Hardy-Weinberg equilibrium. HPA1-5 gene frequencies for Chinese people in Handan were consistent with those of Chinese people in Shijiazhuang (P > 0.05). Among the HPA1-13, -15, the frequencies of HPA-1, -2, -6 for Chinese people in Handan differed appreciably from those for Chinese people in Taiwan (P < 0.05), others were similar to those of Chinese people in Taiwan. Among the HPA 1 - 8, a similarity was noted between Chinese people in Handan and Koreans (P > 0.05), except for HPA-3. Frequencies of HPA-1, -2, -5 significantly were differed from those in African Americans, as compared with HPA 1-5 (P < 0.05). Comparison of gene frequencies from HPA-1 and -5 showed significant differences between Chinese people in Handan and people in UK (P < 0.05). It is concluded that HPA-2, -3, -5, -15 of people in Western region of China have polymorphism, incompatible frequency of HPA antigen distribution is higher, which inevitably results in the increase of immunologic exposure, therefore attention must be paid to the importance of HPA-2, -3, -5, -15 in clinical disorders. This study for the first time completely analyses HPA1-16 gene frequencies in China, and provides data for establishing a typed platelet donor panel in Handan, China.


Subject(s)
Humans , Antigens, Human Platelet , Classification , Genetics , Blood Donors , China , Gene Frequency , Genotype , Platelet Transfusion , Polymerase Chain Reaction , Methods , Polymorphism, Genetic
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