Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 50
Filter
1.
Article in Chinese | WPRIM | ID: wpr-928400

ABSTRACT

OBJECTIVE@#To determine the carrier rate for 21 inherited metabolic diseases among a Chinese population of childbearing age.@*METHODS@#A total of 897 unrelated healthy individuals (including 143 couples) were recruited, and DNA was extracted from their peripheral blood samples. Whole exome sequencing (WES) was carried out to screen potential variants among 54 genes associated with 21 inherited metabolic diseases. Pathogenic and likely pathogenic variants and unreported loss-of-function variants were analyzed.@*RESULTS@#One hundred fourty types of pathogenic/likely pathogenic variants (with an overall number of 183) and unreported loss-of-function variants were detected, which yield a frequency of 0.20 per capita. A husband and wife were both found to carry pathogenic variants of the SLC25A13 gene and have given birth to a healthy baby with the aid of preimplantation genetic diagnosis. The detected variants have involved 40 genes, with the most common ones including ATP7B, SLC25A13, PAH, CBS and MMACHC. Based on the Hardy-Weinberg equilibrium, the incidence of the 21 inherited metabolic diseases in the population was approximately 1/1100, with the five diseases with higher incidence including citrullinemia, methylmalonic acidemia, Wilson disease, glycogen storage disease, and phenylketonuria.@*CONCLUSION@#This study has preliminarily determined the carrier rate and incidence of 21 inherited metabolic diseases among a Chinese population of childbearing age, which has provided valuable information for the design of neonatal screening program for inherited metabolic diseases. Pre-conception carrier screening can provide an important measure for the prevention of transmission of Mendelian disorders in the population.


Subject(s)
Asians/genetics , China , Exome , Female , Humans , Infant, Newborn , Metabolic Diseases/genetics , Mitochondrial Membrane Transport Proteins/genetics , Oxidoreductases/genetics , Whole Exome Sequencing
2.
Journal of Chinese Physician ; (12): 1286-1289, 2021.
Article in Chinese | WPRIM | ID: wpr-909698

ABSTRACT

Objective:To explore the genetic etiology for a premature ovarian insufficiency (POI) patient from a consanguineous Chinese family, and to provide basis for genetic counseling and fertility counseling.Methods:Whole-exome sequencing was performed using DNA extracted from the blood sample of POI patient. Suspected pathogenic mutation was analyzed by bioinformatics methods and verified by Sanger sequencing. The pathogenicity of the variation was assessed according to the ACMG genetic variation classification criteria and guidelines.Results:A homozygous variation, c. 32G>T (p.G11V), of PSMC3IP was identified in the patient. Bioinformatics analysis revealed that the variation was conserved in different animal species, and this variation was classified as possible pathogenic variation according to the ACMG genetic variation classification criteria and guidelines.Conclusions:The homozygous missense variation of PSMC3IP is the cause of the POI patient in this family. We are reporting for the first time the missense variation in PSMC3IP gene caused POI, which enriched the mutation spectrum of PSMC3IP and provided the basis for genetic counseling and fertility guidance of this family.

3.
Article in Chinese | WPRIM | ID: wpr-905921

ABSTRACT

Objective:To control the quality of the reference sample of Wenjingtang by establishing the specific chromatograms. Method:On the basis of analyzing 15 batches of Wenjingtang freeze-dried powder samples, a high performance liquid chromatography (HPLC) specific chromatogram analysis method of Wenjingtang was established. The system adaptability was investigated and the retention time, relative retention value and deviation caused by different chromatographic columns and instruments were calculated by using the same brand of chromatographic columns, four different brands of chromatographic columns and instruments from three different manufacturers. The precision, repeatability and stability of this method was further completed. The possible chemical components of the freeze-dried powders were speculated and identified by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS<italic><sup>n</sup></italic>). Chromatographic separation was performed on ACQUITY UPLC BEH C<sub>18</sub> column (2.1 mm×100 mm, 1.7 μm) with acetonitrile (A)-0.1% formic acid aqueous solution (B) as mobile phase for gradient elution (0-2.8 min, 10%A; 2.8-8.0 min, 10%-18%A; 8.0-12.2 min, 18%-25%A; 12.2-15.3 min, 25%-40%A; 15.3-17.4 min, 40%A; 17.4-20.5 min, 40%-90%A), and column temperature was set at 30 ℃ with flow rate of 0.4 mL·min<sup>-1</sup>. Mass spectrometry was performed on electrospray ionization, data were collected under positive and negative ion modes, and the detection range was <italic>m</italic>/<italic>z</italic> 50-1 600. Result:Ten characteristic peaks were selected as the distinguishing features in this specific chromatograms, and eight of them were identified by comparing with the reference standards, including paeoniflorin (peak 1), liquiritin apioside (peak 2), liquiritin (peak 3), ferulic acid (peak 4), iquiritigenin (peak 6), cinnamaldehyde (peak 8), paeonol (peak 9)and glycyrrhizic acid (peak 10). By mass spectrometry analysis, 30 compounds were identified, and the source of medicinal materials were assigned. It mainly contained triterpenoid saponins and flavonoids from Glycyrrhizae Radix et Rhizoma, ginsenosides from Ginseng Radix et Rhizoma, monoterpenoid glycosides and tannins from Paeoniae Radix Alba, steroids in Achyranthis Bidentatae Radix, phenolic acids in Angelicae Sinensis Radix. Conclusion:The established characteristic chromatographic analysis method of Wenjingtang is simple, stable and repeatable. The chemical composition of the freeze-dried powder of Wenjingtang is basically defined by mass spectrometry identification and source attribution, which can provide reference for the development and quality control of Wenjingtang in the future.

4.
Acta Pharmaceutica Sinica B ; (6): 1885-1902, 2021.
Article in English | WPRIM | ID: wpr-888840

ABSTRACT

Multidrug resistance (MDR) mediated by ATP binding cassette subfamily B member 1 (ABCB1) is significantly hindering effective cancer chemotherapy. However, currently, no ABCB1-inhibitory drugs have been approved to treat MDR cancer clinically, mainly due to the inhibitor specificity, toxicity, and drug interactions. Here, we reported that three polyoxypregnanes (POPs) as the most abundant constituents of

5.
Acta Pharmaceutica Sinica B ; (6): 3820-3835, 2021.
Article in English | WPRIM | ID: wpr-922443

ABSTRACT

Pyrrolizidine alkaloids (PAs) are the most common phytotoxins with documented human hepatotoxicity. PAs require metabolic activation by cytochromes P450 to generate toxic intermediates which bind to proteins and form protein adducts, thereby causing cytotoxicity. This study investigated the role of the gut-liver axis in PA intoxication and the underlying mechanisms. We exposed mice to retrorsine (RTS), a representative PA, and for the first time found RTS-induced intestinal epithelium damage and disruption to intestinal barrier function. Using mice with tissue-selective ablation of P450 activity, we found that hepatic P450s, but not intestinal P450s, were essential for PA bioactivation. Besides, in RTS-exposed, bile duct-cannulated rats, we found the liver-derived reactive PA metabolites were transported by bile into the intestine to exert enterotoxicity. The impact of gut-derived pathogenic factors in RTS-induced hepatotoxicity was further studied in mice with dextran sulfate sodium (DSS)-induced chronic colitis. DSS treatment increased the hepatic endotoxin level and depleted hepatic reduced glutathione, thereby suppressing the PA detoxification pathway. Compared to RTS-exposed normal mice, the colitic mice displayed more severe RTS-induced hepatic vasculature damage, fibrosis, and steatosis. Overall, our findings provide the first mode-of-action evidence of PA-induced enterotoxicity and highlight the importance of gut barrier function in PA-induced liver injury.

6.
Article in Chinese | WPRIM | ID: wpr-921638

ABSTRACT

Based on the textual research on literature, the key information of Wenjing Decoction were tested and identified, and 15 batches of lyophilized powder samples of Wenjing Decoction were prepared. The specific components, including paeoniflorin, glycyrrhizin, ginsenosides(Rg_1, Re and Rb_1), glycyrrhizic acid, and paeonol, were used as indexes to establish the HPLC method for quantitative evaluation, and the content ranges and transfer rates of these components were determined. The results showed that the contents of paeoniflorin, glycyrrhizin, ginsenosides Rg_1 + Re, ginsenoside Rb_1, glycyrrhizic acid, and paeonol in the 15 batches of samples were 0.62%-0.86%, 0.25%-0.76%, 0.14%-0.30%, 0.07%-0.21%, 0.63%-1.16%, and 0.09%-0.25%, respectively, and their transfer rates from the decoction pieces to the reference materials were 14.99%-19.42%, 28.11%-40.93%, 25.92%-61.88%, 25.03%-64.06%, 23.43%-35.53%, and 5.34%-10.44%, respectively. The consistency of the transfer rates between batches indicated that the preparation process was stable. It is suggested that the contents of paeoniflorin, glycyrrhizin, ginsenosides Rg_1 + Re, ginsenoside Rb_1, glycyrrhizic acid, and paeonol in Wenjing Decoction should not be less than 0.52%, 0.35%, 0.15%, 0.10%, 0.63%, and 0.12%, respectively. In this study, we determined the contents and analyzed the quantity transfer process of the index components in Wenjing Decoction, which can provide a basis for the follow-up development of Wenjing Decoction and the quality control of related preparations.


Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Glycyrrhizic Acid , Powders , Quality Control
7.
Asian Journal of Andrology ; (6): 197-204, 2021.
Article in English | WPRIM | ID: wpr-879740

ABSTRACT

Oligoasthenoteratozoospermia (OAT) refers to the combination of various sperm abnormalities, including a decreased sperm count, reduced motility, and abnormal sperm morphology. Only a few genetic causes have been shown to be associated with OAT. Herein, we identified a novel homozygous frameshift mutation in meiosis-specific nuclear structural 1 (MNS1; NM_018365: c.603_604insG: p.Lys202Glufs*6) by whole-exome sequencing in an OAT proband from a consanguineous Chinese family. Subsequent variant screening identified four additional heterozygous MNS1 variants in 6/219 infertile individuals with oligoasthenospermia, but no MNS1 variants were observed among 223 fertile controls. Immunostaining analysis showed MNS1 to be normally located in the whole-sperm flagella, but was absent in the proband's sperm. Expression analysis by Western blot also confirmed that MNS1 was absent in the proband's sperm. Abnormal flagellum morphology and ultrastructural disturbances in outer doublet microtubules were observed in the proband's sperm. A total of three intracytoplasmic sperm injection cycles were carried out for the proband's wife, but they all failed to lead to a successful pregnancy. Overall, this is the first study to report a loss-of-function mutation in MNS1 causing OAT in a Han Chinese patient.

8.
Article in Chinese | WPRIM | ID: wpr-879598

ABSTRACT

OBJECTIVE@#To analyze the (CGG)n repeats of FMR1 gene among patients with unexplained mental retardation.@*METHODS@#For 201 patients with unexplained mental retardation, the (CGG)n repeats of the FMR1 gene were analyzed by PCR and FragilEase@*RESULTS@#For the 201 patients with unexplained mental retardation, 15 were identified with full mutations of the FMR1 gene. The prevalence of fragile X syndrome (FXS) in patients with unexplained mental retardation was determined as 7.5% (15/201). Prenatal diagnosis was provided for 6 pregnant women with pre- or full mutations. Analysis revealed that women with mental retardation and full FMR1 mutations exhibited a skewed XCI pattern with primary expression of the X chromosome carrying the mutant allele.@*CONCLUSION@#FXS has a high incidence among patients with unexplained mental retardation. Analysis of FMR1 gene (CGG)n repeats in patients with unexplained mental retardation can facilitate genetic counseling and prenatal diagnosis for their families. FMR1 gene (CGG)n repeats screening should be recommended for patients with unexplained mental retardation.


Subject(s)
Female , Fragile X Mental Retardation Protein/genetics , Fragile X Syndrome/genetics , Humans , Intellectual Disability/genetics , Mutation , Pregnancy , Prenatal Diagnosis
9.
Article in Chinese | WPRIM | ID: wpr-879582

ABSTRACT

OBJECTIVE@#To explore the correlation between Fragile X mental retardation gene-1 (FMR1) gene CGG repeats with diminished ovarian reserve (DOR).@*METHODS@#For 214 females diagnosed with DOR, DNA was extracted from peripheral blood samples. FMR1 gene CGG repeats were determined by PCR and capillary electrophoresis.@*RESULTS@#Three DOR patients were found to carry FMR1 premutations, and one patient was found to carry gray zone FMR1 repeats. After genetic counseling, one patient and the sister of another patient, both carrying FMR1 permutations, conceived naturally. Prenatal diagnosis showed that both fetuses have carried FMR1 permutations.@*CONCLUSION@#FMR1 gene permutation may be associated with DOR. Determination of FMR1 gene CGG repeats in DOR patients can provide a basis for genetic counseling and guidance for reproduction.


Subject(s)
Female , Fragile X Mental Retardation Protein/metabolism , Fragile X Syndrome/genetics , Humans , Ovarian Diseases , Ovarian Reserve/genetics , Primary Ovarian Insufficiency/genetics , Trinucleotide Repeats/genetics
10.
Journal of Medical Postgraduates ; (12): 454-459, 2020.
Article in Chinese | WPRIM | ID: wpr-821873

ABSTRACT

In the 1970s, the Framingham study found that smoking increased the risk of stroke. Subsequent laboratory and clinical studies have demonstrated that a variety of harmful components in tobacco can damage the vascular endothelium and interfere with the clotting mechanism, giving rise to atherosclerosis and hypercoagulability, which eventually leads to stroke. Furthermore, the harmful ingredients in tobacco may induce hypertension, diabetes, atrial fibrillation and other diseases, which indirectly increase the risk of stroke. However, there are still a lot of questions about the relationship between smoking and stroke. This paper will analyze the correlation between them and its mechanism, and discuss the influence of smoking cessation on the prognosis of stroke, so as to provide reference for the prevention and treatment of related diseases.

11.
Chinese Journal of Dermatology ; (12): 889-893, 2017.
Article in Chinese | WPRIM | ID: wpr-664425

ABSTRACT

Objective To investigate association of vitamin D receptor (VDR) polymorphisms with susceptibility to psoriasis vulgaris and clinical response to calcipotriol in patients with psoriasis vulgaris.Methods A total of 110 patients with psoriasis vulgaris and 183 healthy controls were enrolled into this study,and they were all of Han nationality from Hainan province.Ligase detection reaction (LDR) was conducted to determine the genotypes of VDR gene polymorphisms rs2228570,rs731236,rs1544410 and rs7975232.Single nucleotide polymorphism (SNP)-based association analysis in genotypic and allelic models,and haplotype-based association analysis were then performed.Then,75 patients with psoriasis area and severity index (PASI) scores less than 10 were topically treated with calcipotriol ointment alone.After 6-week treatment,the efficacy of calcipotriol ointment was evaluated,and the correlation between the efficacy and individual genotypes was analyzed.Results The frequency of A allele of rs7975232 in the psoriasis group and control group was 39.09% and 27.05% respectively,and the risk of developing psoriasis in rs7975232 A allele carriers was significantly higher than that in non-carriers (OR =1.731,95% CI:1.213-2.471,P < 0.05).Additionally,the risk of developing psoriasis in individuals with AA genotype (OR =2.404,95% CI:1.085-5.328,P < 0.05),as well as in individuals with AC genotype (OR =2.143,95% CI:1.283-3.579,P < 0.05),was significantly higher than that in patients with CC genotype.CTGA haplotype carriers (rs2228570,rs731236,rs1544410,rs7975232,respectively) had significantly higher risk of developing psoriasis compared with non-carriers (OR =1.907,95% CI:1.132-3.214,P < 0.05).Among 72 patients with mild-to-moderate psoriasis whose PASI scores were less than 10,patients with CC genotype of rs7975232 showed better response to calcipotriol ointment compared with those with AC genotype (OR =3.798,95% CI:1.061-13.590,P < 0.05) and those with AA genotype (OR =9.667,95%CI:1.556-60.040,P < 0.05).Conclusion VDR polymorphisms are associated with psoriasis susceptibility and clinical response to calcipotriol in patients with psoriasis individuals.

12.
Article in Chinese | WPRIM | ID: wpr-657635

ABSTRACT

Objective To explore the clinical effect of acupuncture combined with Chinese Medicine in the treatment of ischemic stroke patients with Qi deficiency and blood stasis restoring stage. Methods 180 patients with ischemic stroke in recovery period of Qi deficiency and blood stasis were divided into control group and study group, 90 cases in each group. The control group received conventional rehabilitation therapy, the study group on the basis of routine rehabilitation therapy combined with traditional Chinese medicine and acupuncture. Recorded the change of nerve function pre-treament and after continuous treatment for 1 month. Results There was no significant difference in neurological function between the two groups before treatment. After treatment, the NIHSS scale scores of the two groups were significantly lower than pre-treatment (P<0.05). After treatment, the improvement effect of nerve function in the study group was better than control group (P<0.05). Conclusion The traditional Chinese medicine and acupuncture on the basis of routine rehabilitation therapy in treating patients ischemic stroke in recovery period of Qi deficiency and blood stasis can get ideal clinical effect.

13.
Article in Chinese | WPRIM | ID: wpr-335101

ABSTRACT

<p><b>OBJECTIVE</b>To explore the genetic etiology of three families affected with split-hand/split-foot malformation (SHFM).</p><p><b>METHODS</b>Peripheral venous blood samples from 21 members of pedigree 1, 2 members of pedigree 2, and 2 members of pedigree 3 were collected. PCR-Sanger sequencing, microarray chip, fluorescence in situ hybridization (FISH), real-time PCR, and next-generation sequencing were employed to screen the mutations in the 3 families. The effect of the identified mutations on the finger (toe) abnormality were also explored.</p><p><b>RESULTS</b>Microarray and real-time PCR analysis has identified a duplication in all patients from pedigrees 1 and 3, which have spanned FKSG40, TLX1, LBX1, BTRC, POLL and FBXW4 (exons 6-9) and LBX1, BTRC, POLL and FBXW4 (exons 6-9) genes, respectively. A missense mutation of the TP63 gene, namely c.692A>G (p.Tyr231Cys), was found in two patients from pedigree 2. FISH analysis of chromosome 10 showed that the rearrangement could fita tandem duplication model. However, next-generation sequencing did not identify the breakpoint.</p><p><b>CONCLUSION</b>The genetic etiology for three families affected with SHFM have been identified, which has provideda basis for genetic counseling and guidance for reproduction.</p>


Subject(s)
Chromosomes, Human, Pair 10 , Genetics , Female , Foot Deformities, Congenital , Genetics , Genetic Testing , Hand Deformities, Congenital , Genetics , Humans , Limb Deformities, Congenital , Genetics , Male , Mutation , Genetics , Pedigree
14.
Article in Chinese | WPRIM | ID: wpr-659918

ABSTRACT

Objective To explore the clinical effect of acupuncture combined with Chinese Medicine in the treatment of ischemic stroke patients with Qi deficiency and blood stasis restoring stage. Methods 180 patients with ischemic stroke in recovery period of Qi deficiency and blood stasis were divided into control group and study group, 90 cases in each group. The control group received conventional rehabilitation therapy, the study group on the basis of routine rehabilitation therapy combined with traditional Chinese medicine and acupuncture. Recorded the change of nerve function pre-treament and after continuous treatment for 1 month. Results There was no significant difference in neurological function between the two groups before treatment. After treatment, the NIHSS scale scores of the two groups were significantly lower than pre-treatment (P<0.05). After treatment, the improvement effect of nerve function in the study group was better than control group (P<0.05). Conclusion The traditional Chinese medicine and acupuncture on the basis of routine rehabilitation therapy in treating patients ischemic stroke in recovery period of Qi deficiency and blood stasis can get ideal clinical effect.

15.
Progress in Modern Biomedicine ; (24): 5389-5391,5395, 2017.
Article in Chinese | WPRIM | ID: wpr-615096

ABSTRACT

Fertilization is a crucial step for origin of life.During Assisted Reproductive Technologies (ART),total fertilization failure is complex and unpredictable.Total fertilization failure may related to some abnormal cellular mechanistic events,such as:any stage of sperm and cumulus-oocyte-complexes penetration,sperm-zona pellucida binding / penetration,sperm-oocyte membrane binding,oocyte activation,sperm discondensation or pronuclear formation.Most of total fertilization failure could be solved by intracytoplasmic sperm injection.But oocytes of some patient still can't fertilize successfully,even though assisted oocyte activation be used.As for total fertilization failure patients in ART,combining the mature of oocyte,sperm quality and some trail to improve clinical protocol in later cycle may prevent failure to happen again.

16.
Progress in Modern Biomedicine ; (24): 4986-4989, 2017.
Article in Chinese | WPRIM | ID: wpr-614914

ABSTRACT

DMD/BMD is a X-linked recessive hereditary disease.It predominantly affects males.While female carriers do not have symptoms,due to their inactive X chromosome make it present mosaic.Recently,more and more papers reported that a clinically significant proportion of DMD/BMD female carriers have symptoms.They presented variable degrees of symptoms.But the mechanism of the pathogencity is still not clear.Most of the research considered that the dominating reason is the skewed X inactivation.It means that the predominant expression of the DMD mutant allele make the normal one have weak expression,thus no function dystrophin proteins could be generate,manifested as DMD/BMD.In this paper,we mainly summarized the relationship between skewed X inactivation and pathogenicity of the symptomatic DMD female carriers.

17.
Article in Chinese | WPRIM | ID: wpr-319656

ABSTRACT

<p><b>OBJECTIVE</b>To study the relationship between morphological characteristics of growth cycle for Epimedium acuminatium and accumulation level of secondary metabolites.</p><p><b>METHOD</b>After making habitat clear, methods of morphology observation, specimen preparation, organism charting were applied to record morphological characteristics of E. acuminatium at different stages. HPLC was employed to detect icariin content of different parts of the plant at the stage of vegetative growth and sexual reproduction.</p><p><b>RESULT</b>The growth cycle of E. acuminatium was divided into vegetative growth stage and sexual and asexual reproduction coexistence stage. Seven to eight years were needed for the plants flowering and seeding of E. acuminatimcan in open land of forest edge, but within good nutrition conditions, growth cycle is shortened into three to four years. The difference of icariin content in two growth stages was not significant.</p><p><b>CONCLUSION</b>Morphological change of lower and foliar organ can be utilized to estimate growth age. But after applying sexual propagation for both root and stem of grown plants, it's been difficult to determine their growth year. If individual biomass or yield is economical, plants can be harvested in both vegetative growth and sexual reproduction stages.</p>


Subject(s)
Biomass , Chromatography, High Pressure Liquid , Methods , Epimedium , Metabolism , Flavonoids , Metabolism , Plant Roots , Metabolism , Reproduction , Seedlings , Metabolism
18.
Article in Chinese | WPRIM | ID: wpr-814616

ABSTRACT

OBJECTIVE@#To investigate the effectiveness of treatment for severe ovary hyperstimulation syndrome (OHSS) complicated by pleural effusion and ascites after in vitro fertilization preembryo transfer (IVF-ET).@*METHODS@#One hundred and thirty-two patients with severe OHSS in our hospital (from January 2007 to December 2010) were retrospectively analyzed and the efficacy of three therapeutic methods was compared. Twenty-five patients in group I were treated with low-molecular dextran and albumin, 67 patients in group II were treated with 6% medium molecular-weight hydroxyethyl starch, and 40 patients in group III were treated with active aspiration of pleural effusion and ascites.@*RESULTS@#All three therapies improved the symptoms of OHSS and various blood biochemical parameters. The duration of hospitalization of group III [(7.4±4.5) d] was significantly less than those of group I [(21.4±9.2) d] or II [(15.6±6.7) d], and the cost of group III [(2656.2±1882.8) Yuan] was also significantly lower than that of group I or II [(11937.6±7989.8) and (5182.7±2991.7) Yuan, respectively].@*CONCLUSION@#Abdominal B ultrasonography-guided trans-abdominal wall aspiration of pleural effusion and ascites combined with blood volume maintenance is an effective and economical way to treat OHSS.


Subject(s)
Adult , Ascites , Therapeutics , Dextrans , Drainage , Female , Fertilization in Vitro , Humans , Hydroxyethyl Starch Derivatives , Infusions, Intravenous , Ovarian Hyperstimulation Syndrome , Therapeutics , Ovulation Induction , Pleural Effusion , Therapeutics , Retrospective Studies , Serum Albumin
19.
Article in Chinese | WPRIM | ID: wpr-315528

ABSTRACT

<p><b>OBJECTIVE</b>To establish a human embryonic stem cell line with stably β-catenin gene silencing by lentivirus-mediated shRNA interference.</p><p><b>METHODS</b>PLKO.1-puro-β-catenin vector, a lentivirus plasmid expressing β-catenin shRNA, was packaged into 293FT cells. Human embryonic stem cells were infected with the lentivirus and the cell clones stably expressing β-catenin shRNA were selected by puromycin, with the uninfected cells and cells infected with the empty vector as the control. Real-time RT-PCR was used to evaluate the efficiency of β-catenin knocked down; β-catenin and OCT4 protein expression in the infected cells was examined using immunofluorescence assay.</p><p><b>RESULTS</b>Infection with β-catenin-specific shRNA lentivirus resulted in stable interference of β-catenin expression in human embryonic stem cells, which showed a β-catenin mRNA expression of only 1% of that in the uninfected cells. Infection with the empty vector produced no obvious effect on β-catenin expression compared to the uninfected cells. In the cells infected with β-catenin shRNA lentivirus, β-catenin protein expression was almost undetectable in immunofluorescence assay, while OCT4 was still expressed after the interference.</p><p><b>CONCLUSION</b>Lentiviral vector-delivered shRNA results in effective and stable β-catenin gene silencing in human embryonic stem cells.</p>


Subject(s)
Cell Line , Embryonic Stem Cells , Cell Biology , Metabolism , Genetic Vectors , Humans , Lentivirus , Genetics , Plasmids , RNA Interference , RNA, Small Interfering , Genetics , Transfection , Wnt Signaling Pathway , beta Catenin , Genetics , Metabolism
20.
Article in Chinese | WPRIM | ID: wpr-358743

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of living high-training low (HiLo) on innate immunity in blood of elite swimmers.</p><p><b>METHODS</b>Six female swimmers undertook HiLo for two weeks, erythrocyte adhesion function and counts of leukocyte were tested in different time of training period.</p><p><b>RESULTS</b>Red blood cell C3b receptor ring rate (RBC-C3bRR) decreased and red blood cell immune complex matter ring rate (RBC-ICR) increased significantly (P < 0.05), the two markers returned to base line 1 week after training. Counts of leukocyte and granulocyte decreased significantly (P < 0.05), and they recovered 1 week after training; Counts of lymphocyte and monocyte decreased without significance during training and did not recovered after training.</p><p><b>CONCLUSION</b>Immunity of erythrocyte and granulocyte decreased quickly, but lymphocyte and monocyte recovered slowly, swimmers were adaptive to the training.</p>


Subject(s)
Adolescent , Altitude , Athletes , Cell Adhesion , Erythrocytes , Cell Biology , Allergy and Immunology , Female , Humans , Leukocyte Count , Physical Education and Training , Swimming
SELECTION OF CITATIONS
SEARCH DETAIL