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@#AIM: To evaluate the changes of early visual quality of patients with high myopia after small incision lenticule extraction(SMILE)and femtosecond assisted laser in situ keratomileusis(FS-LASIK)by the double-pass optical quality analysis system Ⅱ(OQAS Ⅱ)and Pentacam corneal topography.<p>METHODS: A prospective non-randomized controlled study was conducted among 148 eyes of 74 patients with high myopia. These patients were treated by the same surgeon in our hospital from March 2020 to September 2020. According to their wishes, 86 eyes with 43 patients were treated with SMILE and 62 eyes with 31 patients were treated with FS-LASIK. The uncorrected visual acuity(UCVA), modulation transfer function cut off frequency(MTF cut off), Strehl ratio(SR), objective scattering index(OSI), predicted visual acuity values(VA 100%, VA 20%, VA 9%), high-order aberration(HOA), horizontal aberration(<i>Z 13</i>), vertical coma(<i>Z -13</i>)and spherical aberration(<i>Z 04</i>)preoperatively, 1 and 7d postoperatively were collected and analyzed.<p>RESULTS:There was no statistical significance in preoperative age, spherical equivalent(SE), UCVA, MTF cut off, SR, OSI, VA 100%, VA 20%, VA 9%, HOA, <i>Z 13</i>, <i>Z -13</i> and <i>Z 04</i>(<i>P</i>>0.05). The OSI of the SMILE group was higher than the FS-LASIK group at postoperative 1d(2.3±2.1 <i>vs</i> 1.8±1.1). The difference in OSI between the two groups was not statistically significant at postoperative 7d(1.2±0.7 <i>vs</i> 1.3±0.7). The HOA and <i>Z04</i> in the FS-LASIK group were higher than the SMILE group at postoperative 1d and 7d(<i>P</i><0.001). The UCVA, <i>Z 13</i>, <i>Z -13</i>, MTF cut off, SR, VA 100%, VA 20%, and VA 9% between the two groups were no statistical significance at postoperative 1d and 7d(<i>P</i>>0.05).<p>CONCLUSION:FS-LASIK is easier to introduce corneal high-order aberration and spherical aberration in the early postoperative period, while SMILE with 2mm incision only introduces higher scatter at postoperative 1d. Therefore, SMILE can obtain better visual quality than FS-LASIK in the early postoperative period among patients with high myopia after corneal refractive surgery.
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Cancer-related anorexia syndrome (CACS) is one of the main causes for death at present as well as a syndrome seriously harming patients' quality of life, treatment effect and survival time. In current clinical researches, there are fewer reports about empirical traditional Chinese medicine(TCM) prescriptions and patent prescriptions treating CACS, and prescription rules are rarely analyzed in a systematic manner. As the hidden rules are not excavated, it is hard to have an innovative discovery and knowledge of clinical medication. In this paper, the grey screening method combined with the multivariate statistical method was used to build the ″CACS prescriptions database″. Based on the database, totally 359 prescriptions were selected, the frequency of herbs in prescription was determined, and commonly combined drugs were evolved into 4 new prescriptions for different syndromes. Prescriptions of TCM in treatment of CACS gave priority to benefiting qi for strengthening spleen, also laid emphasis on replenishing kidney essence, dispersing stagnated liver-qi and dispersing lung-qi. Moreover, interdependence and mutual promotion of yin and yang should be taken into account to reflect TCM's holism and theory for treatment based on syndrome differentiation. The grey screening method, as a valuable traditional Chinese medicine research-supporting method, can be used to subjectively and objectively analyze prescription rules; and the new prescriptions can provide reference for the clinical use of TCM for treating CACS and the drug development.
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The compatibility law analysis of traditional Chinese medicine (TCM) is the core of modernization of TCM. Furthermore, "macro compatibility" and "micro compatibility" are two important components of formula compatibility. At present, due to the lack of research on the law of "macro compatibility", the research ideas are too narrow, which seriously restricts the development of modernization of Chinese materia medica (CMM). Under the guidance of TCM theory, we explore the study on the new mode of "grey screening" whose components are divided into the functional units by the current situation and deficiency of CMM research, treatment based on syndrome differentiation, grey systematology, the action of multivariate statistical methods, and new idea on the feasibility study and application prospect as well.
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OBJECTIVE@#To investigate the impact of bone marrow mesenchymal stem cells on Smad expression of hepatic fibrosis rats.@*METHODS@#A total of 48 adult female SD rats were randomly divided into three groups, normal control group (n=10), observation group (n=19) with liver fibrosis model rats injected with BMSCs cells; model group (n=19), with liver fibrosis model rats injected with physiological saline. Serum index, TGF-β1 and Smad expression were detected.@*RESULTS@#Type III procollagen, IV collagen, hyaluronic acid, laminin levels of observation group were significantly lower than those of model group (P<0.05). The content and expression of TGF-β1 in serum and liver tissue of observation group were significantly lower than those of model group(P<0.05). Compared with normal control group, the Smad3, Smad4 mRNA and protein expression of model group were significantly increased, the Smad7 mRNA and protein expression were significantly reduced (P<0.05). Compared with model group, Smad3, Smad4 mRNA and protein expression of observation group were significantly reduced, and Smad7 mRNA expression were significantly increased (P<0.05).@*CONCLUSIONS@#BMSCs can regulate Smad expression to some extent, and reduce the degree of liver fibrosis.
Subject(s)
Animals , Female , Rats , Bone Marrow Cells , Cell Biology , Collagen , Metabolism , Hyaluronic Acid , Metabolism , Laminin , Metabolism , Liver Cirrhosis , Metabolism , Pathology , General Surgery , Mesenchymal Stem Cell Transplantation , Methods , Mesenchymal Stem Cells , Rats, Sprague-Dawley , Smad Proteins , Metabolism , Transforming Growth Factor beta , MetabolismABSTRACT
OBJECTIVE@#To observe the effect of Rougan Huaqian granules combined with human mesenchymal stem cell (hMSC) transplantation on the liver fibrosis in carbon tetrachloride-induced cirrhosis rats.@*METHODS@#Sixty SD rats were randomly divided into five groups. The rats in control group received intraperitoneal injection of saline, while those in model control group, treatment group A, group B and group C received intraperitoneal injection of carbon tetrachloride oily solution to induce liver cirrhosis within 8 weeks. Then, the rats in the model control group, treatment group A, treatment group B, treatment group C received vein tail injection of saline, Rougan Huaqian granules, hMSC suspension and Rougan Huaqian granules combined with hMSC suspension.@*RESULTS@#The treatment groups had significantly different liver function (AST levels), liver fibrosis index (laminin and HA), hepatic sinusoidal wallsα-smooth muscle actin, IV collagen and laminin protein expression and I, III collagen from the model group (P<0.05). The transplanted cells showed human hepatocyte-like cells differentiation trend in the liver.@*CONCLUSIONS@#The Rougan Huaqian granules combined with hMSC transplantation can alleviate liver fibrosis in cirrhosis rats.
Subject(s)
Animals , Female , Humans , Male , Rats , Body Weight , Collagen , Genetics , Metabolism , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Liver , Chemistry , Metabolism , Pathology , Liver Cirrhosis , Metabolism , Pathology , Therapeutics , Matrix Metalloproteinase 13 , Genetics , Metabolism , Mesenchymal Stem Cell Transplantation , Organ Size , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects and safety of Western medicine combined with Chinese medicine (CM) based on syndrome differentiation in the treatment of elderly polarized hypertension (PHPT), or isolated systolic hypertension with low diastolic blood pressure (DBP).</p><p><b>METHODS</b>A total of 125 elderly patients with PHPT were randomly assigned to two groups: 59 in the control group treated by Western medicine and 66 in the intervention group treated by Western medicine combined with CM treatment. Based on syndrome differentiation, the patients in the intervention group were further divided into subgroups of yang-qi deficiency and yin-qi deficiency. All subjects were treated with Western medicine of Amlodipine Besylate Tablets and Irbesartan Tablets (or Irbesartan and Hydrochlorothiazide Tablets), to decrease their systolic blood pressure (SBP) slowly to 125-135 mm Hg in 2-6 weeks. In the intervention group, Shiyiwei Shenqi Capsule was given additionally to the subgroup of yang-qi deficiency at the dosage of 3-5 capsules, thrice a day, while Dengzhan Shengmai Capsule was given additionally to the subgroup of yin-qi deficiency at the dosage of 2 capsules, 2-3 times per day. For all subjects, SBP, pulse pressure (PP), and DBP were measured before treatment and at the terminal of a 6-week treatment. For subjects in the intervention group, left ventricular ejection fraction (LVEF) was also recorded.</p><p><b>RESULTS</b>After a 6-week treatment, the SBP in the two groups and the PP in the intervention group decreased significantly compared to those before treatment (P<0.05), while the PP in the control group showed no significant difference between prior and post-treatment (P>0.05). After treatment, the DBP in the control group decreased (P>0.05), while the DBP and LVEF in the intervention group showed an increase tendency although it had no statistical significance (P>0.05). When subjects in the intervention group were classified further by the course of disease, the DBP and LVEF of subjects whose course of disease were less than 2 years, increased significantly after treatment (P<0.05).</p><p><b>CONCLUSION</b>Western medicine combined with CM treatment based on syndrome differentiation was safer and more effective than Western medicine alone in the treatment of elderly PHPT, because it not only reduced SBP but also improved DBP, which might lower the incidence of the cardiovascular and cerebrovascular events.</p>
Subject(s)
Aged , Female , Humans , Male , Amlodipine , Pharmacology , Therapeutic Uses , Antihypertensive Agents , Pharmacology , Therapeutic Uses , Biphenyl Compounds , Pharmacology , Therapeutic Uses , Blood Pressure , Diastole , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Hypertension , Drug Therapy , Stroke Volume , Syndrome , Tetrazoles , Pharmacology , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To study the effect of strengthening Pi and nourishing Shen therapy (SPNST) in treating patients with glucocorticoid resistant myasthenia gravis (GR-MG).</p><p><b>METHODS</b>Twenty-seven patients with MG were enrolled, who were relapse cases after treated by cholinesterase inhibitor with systemic glucocorticoid treatment and showed resistance to glucocorticoid. All were treated by Western medicines, methylprednisolone (MP) and pyridostigmine bromide (PSB), together with Chinese medicine (CM) given according to their syndrome types, namely, for the 15 patients of Pi-Shen qi-yin deficiency type, Buzhong Yiqi Pill and Liuwei Dihuang Pill, and for the 12 patients of Pi-Shen yang-deficiency type, Buzhong Yiqi Pill and Zishen Yutai Pill. The dosages of medicines were reduced gradually in MP-PSB-CM order along with the progressing of the therapy in 4 stages (symptom curing, choline receptor restoration, immune regulation, and functional strengthening). Muscle strength and overall state of patients were re-examined before and after each of the 4 stages.</p><p><b>RESULTS</b>After 1-year treatment, the therapeutic effect in 9 patients was judged as completely remitted; in 7 as remitted with continuous medication; in 5, 1 and 3 as significantly improved, moderately improved and unchanged respectively, while 2 patients died.</p><p><b>CONCLUSION</b>Integrative medicine shows definite effects in treating GR-MG, and it is worthy of further studying.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Drug Resistance , Drugs, Chinese Herbal , Therapeutic Uses , Glucocorticoids , Therapeutic Uses , Integrative Medicine , Methods , Myasthenia Gravis , Drug Therapy , Yin Deficiency , Drug TherapyABSTRACT
Objective To construct a recombinant plasmid expressing human microdystrophin gene fused with VP22 ofhmnan herpes simplex virus 1(HSV-1),and investigate the expression of microdystrophin in vitro and the characteristics of VP22-mediated protein transduction.Methods Full length HSV-1 VP22 gene was amplified by PCR from the plasmid pSINrep5-VP22 and cloned into the eukatyotic expression vector pAVXl to conslruct recombinant plasmid pAVP22.Microdystrophin cDNA was obtained from the recombinant plasmid pBSK-MICRO digested with Not Ⅰ,and the product Was inserted into plasmid pAVP22 to construct the plasmid pAVP22-MICDYS. 3T3 cells were transfected with pAVP22-MICDYS,and the expression of microdystrophin was detected by RT-PCR,Western blotting and immtmocytochemislry.The supematant of 3T3 cells transfected with pAVP22-MICDYS were collected to infect human mesenchymal cells(MSCs),and the expression of the fusion protein VP22-microdystrophin in the cells was detected using immunohistochemistry to assess VP22-mediated protein transduction. Results The recombinant plasmid expressing microdystrophin-VP22 fusion gene capable of in vitro expression of the fusion protein was constructed successfully.VP22 was shown to enhance the expression efficiency of microdystrophin in 3T3 cells and transduce VP22-microdystrophin fusion protein from 3T3 cells to human MSCs. Conclusion The recombinant plasmid expressing microdystrophin-VP22 fusion gene with protein transduction capacity provides an important basis for further study of the fusion protein in treatment of Duchenne muscular dystrophy.
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<p><b>OBJECTIVE</b>To investigate the use of surface electromyography (sEMG) and electrocardiogram (ECG) in evaluation of dynamic workload.</p><p><b>METHODS</b>Through controlling the speed and gradient, 8 subjects ran on the treadmill power machine to simulate the dynamic work. The sEMG signal of anterior tibial muscle (AT) and gastrocnemius muscle (GC) of right lower limb and ECG signal were recorded. The root mean square value (RMS), median frequency (MF), mean power frequency (MPF), heart rate (HR), standard deviation of all normal to normal intervals (SDNN) and Borg scores were analyzed.</p><p><b>RESULTS</b>In the five sports, with the speed increasing, all the values of RMS increased in the AT and GC (P < 0.01). With the gradient increasing, the values of RMS increased in the GC (P < 0.01) while the values of RMS of AT had a trend of decrease (P > 0.05). In all five sports, both the values of MF and MPF in AT and GC were lowest in B sports. Compared to A sport, most of the values of MF and MPF increased in C, D, E sports (P < 0.01), with a highest value in the D sport. Compared with A sport, the HR of B, C, D, E sports significantly increased (P < 0.01), and the highest heart rate was found in B sport, however, the values of SDNN significantly decreased. With the increased speed and gradient, the scores of Borg scale significantly increased.</p><p><b>CONCLUSION</b>In the evaluation of dynamic workload, RMS and HR appear to be good indexes. However, in terms of stress reaction to dynamic workload, MF and MPF are more sensitive.</p>
Subject(s)
Adult , Humans , Male , Electrocardiography , Electromyography , WorkloadABSTRACT
<p><b>OBJECTIVE</b>To explore the structural and functional changes of dystrophin molecule after exon 3 deletion.</p><p><b>METHODS</b>Three-dimensional models of dystrophin comprising the major domains were established before and after exon 3 deletion using SWISS-MODEL server. The motifs and structural domains of dystrophin after exon 3 deletion were searched in Pfam database, and the crystal structure of the actin-binding domain in the dystrophin molecule was analyzed using Rasmol software.</p><p><b>RESULTS</b>Torsion of the N-terminal actin-binding domain occurred in the dystrophin molecule after deletion of exon 3. Homology analysis based on Pfam database searches indicated that following exon 3 deletion, the Bit score of the first calponin homology (CH1) domain was decreased from 108 to 36.5 while its expectation value increased from 2.3e-9 to 8.1e-8. The deletion also resulted in the absence of the spiral region C from the CH1 domain.</p><p><b>CONCLUSION</b>Exon 3 deletion in the dystrophin-coding sequence decreases the stability of CH1 domain and prevents the formation of the junction interface where dystrophin binds to actin. The bioinformatics approach provides a new alternative for investigation of the pathogenesis of DMD pathogenesy investigation.</p>
Subject(s)
Humans , Dystrophin , Chemistry , Genetics , Metabolism , Exons , Genetics , Models, Molecular , Muscular Dystrophy, Duchenne , Genetics , Metabolism , Protein Binding , Protein Conformation , Protein Structure, Tertiary , Sequence Deletion , Structure-Activity RelationshipABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of Hg2+ on voltage-dependent calcium channels and intracellular free calcium in trigeminal ganglion neurons of rats and explore the toxicity mechanism of Hg2+ on these neurons.</p><p><b>METHODS</b>Whole cell patch-clamp technique was used to determine ICa of voltage-dependent calcium channels in trigeminal ganglion neurons of rats. Intracellular free calcium was measured to explore [Ca2+]i dynamic changes from a single cell level by laser scanning confocal microscopy and fluorescence probe techniques.</p><p><b>RESULTS</b>0.01, 0.10, 1.00 and 10.00 micromol/L Hg2+ could reduce voltage-dependent calcium channel currents ICa by (1.80+/-0.32)%, (23.04+/-9.46)%, (58.20+/-7.90)% and (82.00+/-5.77)% in trigeminal ganglion neurons. The inhibiting effects reached their maximum in 5 minutes and could not be reversed significantly during wash with Hg2+-free solution. Also, 0.01, 0.10 and 1.00 micromol/L Hg2+ increased intracellular free calcium concentrations by (2.50+/-0.83)%, (82.81+/-35.36)% and (222.70+/-62.48)% in trigeminal ganglion neurons. Pre-administrated trigeminal ganglion neurons with nifedipine for 10 minutes could decrease the effects and delay the effecting time.</p><p><b>CONCLUSION</b>The inhibition of Hg2+ on the voltage-dependent calcium channel currents ICa depends on voltage-dependent calcium channels. And the increase of intracellular free calcium concentration in trigeminal ganglion neurons induced by Hg2+ is related to the release of intracellular stored calcium. However, the relationship between them needs further investigation.</p>
Subject(s)
Animals , Female , Male , Rats , Calcium , Metabolism , Calcium Channels , Metabolism , Physiology , Cells, Cultured , Mercury , Pharmacology , Rats, Sprague-Dawley , Trigeminal Ganglion , Cell Biology , Metabolism , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To detect the female carriers from the intron and/or exon-deletion Duchenne/Becker musclular dystrophy (DMD) familial members for prenatal or preimplantation genetic diagnosis.</p><p><b>METHODS</b>Using method of PCR to five microsatellite markers (located in 5' terminus and intron 44, 45, 49, 50), analysing of the short tandem repeat sequence polymorphism with the genescan and binding with the quantitative polymerase chain reaction, we detected the DMD carriers from 1 intron and exon -deletion family and 1 intron-deletion family.</p><p><b>RESULTS</b>The STR-50 genotype of II 2 in family 5 was 245/245, so II3 is DMD gene carrier. The STR-45 genotype of II6 and II8 were del/172, III19 was del/178, so they were all DMD gene carriers.</p><p><b>CONCLUSION</b>The STR haploid linkage analysis combined with quantitative polymerase chain reaction is accurate and efficient to detect the female carriers from the intron and/or exon-deletion DMD familial members.</p>
Subject(s)
Female , Humans , Male , Exons , Genetics , Gene Deletion , Heterozygote , Introns , Genetics , Microsatellite Repeats , Genetics , Muscular Dystrophy, Duchenne , Diagnosis , Genetics , Pedigree , Polymerase Chain Reaction , MethodsABSTRACT
<p><b>OBJECTIVE</b>To investigate the dynamic changes of dystrophin expression in mdx mice after bone marrow stem cells transplantation.</p><p><b>METHODS</b>The bone marrow stem cells of C57 BL/6 mice (aged 6 to 8 weeks) were injected intravenously into the mdx mice (aged 7 to 9 weeks), which were preconditioned with 7Gy gamma ray. The amount of dystrophin;expression in gastrocnemius was detected by immunofluorescence, reverse transcription-polymerase chain reaction and Western blot at week 5, 8, 12 and 16 after transplantation.</p><p><b>RESULTS</b>At week 5 after bone marrow stem cells transplantation, the dystrophin expression detected in mdx mice were very low; however, its expression increased along with time. At week 16 week, about 12% muscle cells of all transplanted mice expressed dystrophin. There were less centrally placed myonuclei than the control mdx mice, whereas peripheral myonuclei increased.</p><p><b>CONCLUSIONS</b>After having been injected into mdx mice, the allogenic bone marrow stem cells have a trend to reach the injured muscle tissues and differentiate to fibers that can express dystrophin and the expression increased with time. The bone marrow stem cells participates in the repair and regeneration of the injured tissues permanently and constantly.</p>
Subject(s)
Animals , Male , Mice , Bone Marrow Cells , Cell Biology , Metabolism , Cell Differentiation , Disease Models, Animal , Dystrophin , Hematopoietic Stem Cell Transplantation , Mice, Inbred C57BL , Mice, Inbred mdx , Muscular Dystrophy, Duchenne , Metabolism , General Surgery , Transplantation, HomologousABSTRACT
<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector of human microdystrophin gene and observe its expression in rat mesenchymal stem cells (rMSCs) in vitro.</p><p><b>METHODS</b>The plasmid PBSK-MICRO containing human microdystrophin cDNA was digested by restriction endonuclease, and the resultant microdystrophin fragment was inserted into the NotI site of pcDNA3.1(+) to prepare the eukaryotic expression vector-pcDNA3.1(+)/ microdystrophin, which was identified by endonuclease digestion and sequencing. The recombinant plasmid was transfected into rMSCs via lipofectamine, and after G418 selection, the expression of microdystrophin was detected by RT-PCR and indirect immunofluorescence assay.</p><p><b>RESULTS</b>Microdystrophin gene fragment was correctly inserted into the plasmid pcDNA3.1(+), as conformed by sequencing and digestion with Not I and Hind III. The total mRNA of the transfected rMSCs was extracted and microdystrophin mRNA expression was found in the cells by RT-PCR. Indirect immunofluorescence assay for the protein expression of microdystrophin showed bright red fluorescence in the transfected rMSCs.</p><p><b>CONCLUSION</b>Eukaryotic expression plasmid pcDNA3.1(+)/microdystrophin has been constructed successfully and microdystrophin can be expressed in transfected rMSCs in vitro, which may facilitate further research of Duchenne muscular dystrophy treatment by genetically modified allogeneic stem cell transplantation.</p>
Subject(s)
Animals , Humans , Rats , Base Sequence , Cells, Cultured , Dystrophin , Genetics , Fluorescent Antibody Technique, Indirect , Gene Expression , Mesenchymal Stem Cells , Cell Biology , Metabolism , Molecular Sequence Data , Peptide Fragments , Genetics , Plasmids , Genetics , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To detect the disease-causing point mutations in the dystrophin gene of Duchenne muscular dystrophy (DMD) patients.</p><p><b>METHODS</b>The approach of denaturing high performance liquid chromatography (DHPLC) coupling with sequencing was used to screen the point mutations of 79 exons and the untranslated regions of dystrophin gene without large deletions/duplications, which was in 6 unrelated DMD probands from 6 DMD families.</p><p><b>RESULTS</b>Five disease-causing mutations, 697-698insGT, C616T, G1255T, C4279T, and C2302T, were ides created the new stop codons in downstream sites of mutations, respectively. In addition to the disease-causing point mutations, a point mutation T5586+61A in intron 39 was also found at patient 3, and a missense mutation A694T in exon 8 was detected at patient 5. Four point mutations, C2168+13T, 5740-13dupG, G5234A and C5280T, were also detected at patient 6 whose causative point mutation was unavailable. Seven point mutations have not been reported previously. Bi-directional PCR amplification of specific alleles (Bi-PASA) method was established to distinguish the haplotypes of heterozygote or homozygote in a single PCR reaction.</p><p><b>CONCLUSION</b>Via automated DHPLC screening or detecting the subexonic mutations in dystrophin gene is feasible to clinical laboratories, and also is a superior method in terms of sensitivity and efficiency.</p>
Subject(s)
Humans , Male , Base Sequence , Chromatography, High Pressure Liquid , DNA Mutational Analysis , Dystrophin , Genetics , Gene Duplication , Muscular Dystrophy, Duchenne , Genetics , Point Mutation , Polymerase Chain Reaction , Sequence DeletionABSTRACT
<p><b>OBJECTIVE</b>To set up a technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene, and to evaluate the possibility of using this technique for preimplantation genetic diagnosis(PGD) of deleted Duchenne muscular dystrophy (DMD) with family history.</p><p><b>METHODS</b>Fifty single lymphocytes of a normal male and fifty of a normal female were obtained for detecting dystrophin gene(exon 51, exon 19, exon 48) and SRY gene by 3-plex nested PCR.</p><p><b>RESULTS</b>In the group of exon 51/exon 19/SRY, the amplification rates of exon 51, exon 19 and SRY in male were 96%, 94% and 94%; the amplification rates of exon 51 and exon19 in female were 94% and 94%, respectively. In the exon 48/exon 19/SRY group, the amplification rates of exon 48, exon 19 and SRY in male were 92%, 90% and 94%, the amplification rates of exon 48, exon 19 in female were 94% and 92%, respectively.</p><p><b>CONCLUSION</b>The technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene established in this study is highly sensitive, specific and reliable, and is suitable for PGD of deleted DMD with family history.</p>
Subject(s)
Female , Humans , Male , Dystrophin , Genetics , Exons , Genetics , Polymerase Chain Reaction , Methods , Preimplantation Diagnosis , Methods , Reproducibility of Results , Sequence Deletion , Sex Determination ProcessesABSTRACT
<p><b>OBJECTIVE</b>To screen and detect the female carriers from the DMD/BMD family members for prenatal or preimplantation genetic diagnosis.</p><p><b>METHODS</b>For the detection of DMD/BMD carriers from 27 family members in 4 families, PCR to five microsatellite markers(located in 5' terminus and intron 44, 45, 49, 50) and analysis of the short tandem repeat(STR) sequence polymorphism with the use of genescan were implemented.</p><p><b>RESULTS</b>Six of the 17 female members were obligate DMD gene carriers according to the haplotype analysis of the results of the genescan, which conformed with the pedigree analysis. Besides, the authors detected five carriers and five normal females in these families with the use of the haplotype analysis only. The most polymorphic locus was STR 49, and the least was STR 50.</p><p><b>CONCLUSION</b>The STR haploid linkage analysis using (CA)n repeats within the human dystrophin gene is a rapid,accurate, objective method and is well suited for routine use in clinical laboratories engaged in DMD/BMD linkage analysis for the detection of carrier.</p>
Subject(s)
Female , Humans , Male , Genetic Carrier Screening , Microsatellite Repeats , Muscular Dystrophy, Duchenne , Genetics , Polymerase Chain Reaction , Tandem Repeat SequencesABSTRACT
<p><b>OBJECTIVE</b>Study the improvement of locomotive faculty of dystrophin/utropin gene double knock-out mice (dko mice) by transplanting bone marrow stem cells.</p><p><b>METHODS</b>The bone marrow stem cells of C57BL/6 mice (4- to 5-weeks age) were cultured in vitro for three days, before transplanted intravenously (1.0 x 10(7) for each) into 11 dko mice (7- to 8-weeks age). The dko mice were irridiated with 7Gy gamma-ray before transplantation. 8-9 weeks after transplantation, the locomotroy function, electromyography items and expression of dystrophin in transplanted mice and controls were observed.</p><p><b>RESULTS</b>8-9 weeks after transplantation, the dropping times of hauling wire were 3.09 +/- 2.47, compared with that of the control dko mice(16.78 +/- 3.60), there are distinct differences. About electromyography items, the duration of active potential and amplitude of maxim contractions were (4.99 +/- 1.62) ms and(2872 +/- 1474.33) microV, compare with those of control dko mice(3.69 +/- 0.40) ms and(1210.0 +/- 551.0) microV, respectively, about 7% fibers of the muscle tissue of transplanted dko mice expressed dystrophin protein.</p><p><b>CONCLUSIONS</b>8-9 weeks after transplanted with homology bone marrow stem cells, the locomotive function and electromyography items of transplanted dko mice were obviously improved, and about 7% muscle tissue fibers of the mice expressing dystrophin protein were observed. It suggested that there is an ideal prospect for DMD therapy with bone marrow stem cells transplantation.</p>