Objective To investigate the optimal time and molecular mechanism of gene therapy for promoting distraction osteogenesis by observing the effect of gene transfecting at different time on expression of cyclins in mandibular distraction area.Methods Forty eight New-Zealand rabbits were employed.After accomplishing bilateral mandibular distraction osteogenesis model,the rabbits were randomly divided into the group A,B,C and D.The group A,B and C were transfected by recombinant plasmids pIRES-hBMP2-hVEGF165 2 μg(0.1 μg/μL)at the distraction area instantly after operation,on postoperative 4,14 d and given local electroporation stimulation.The four groups entered the consolidation stage after 10 d continuous traction at a rate of 1 mm/d on postoperative 4 d.Three rabbits in each group were respectively sacrificed on 7,14,28,56 d of the consolidation stage.The expression of cyclin A,D1,and E of fresh bone tissue in distraction area were detected by immunohistochemical staining.Results Cyclin A,D1,and E were strongly expressed in the traction gap on 7,14 d of consolidation stage found,which was strongest on 7 d,moreover the group A,B and C were stronger than the group D.The expression in each group was weakened on 28,56 d.The expression on 7 d in the group B was stronger than that in the4 group A,C and D(P<0.05),the expression had no statistical difference between the group A and C(P>0.05),but all were stronger than the group D(P<0.05);the expression in the group B and C on 14 d was stronger than that in the group A and D,but the expression had no statistical difference between the group B and C and between the group A and D(P>0.05).Conclusion The high expression of cyclin A,D1 and E can promote the cellular division,proliferation and differentiation in distraction area,thus accelerates the new bone formation in the distraction area,prompting that the distraction period is the best time for distraction osteogenesis under gene therapeutic intervention.

To determine the effects of Von Hippel-Lindau (VHL) on the invasion and migration of glioma U251 cells. Methods:U251 GBM cells were transfected using VHL expression plasmid. Real-time polymerase chain reaction was conducted to de-tect VHL mRNA expression after transfection. Western blot assay was used to measure protein (VHL, MMP-2, and MMP-9) expres-sion. Tumor invasion and migration were examined by the Transwell and wound-healing experimental methods after VHL up-regula-tion. The intracranial model of nude mouse was developed using U251 cells transfected by VHL expression plasmid, and immunohisto-chemical staining was used to measure protein (VHL, MMP-2, and MMP-9) expression in the tissue sections. Results: In the U251 cells transfected by VHL expression plasmid, the expression of VHL mRNA and VHL proteins increased, and the expression of MMP-2 and MMP-9 protein decreased. Meanwhile, the invasion and migration of glioma U251 cells were also inhibited. Immunohistochemical staining results showed that the expression of MMP-2 and MMP-9 proteins decreased, and the VHL protein expression increased after transfection. Conclusion:VHL can inhibit the invasion and migration of glioma U251 cells. Thus, VHL gene can be used as a target for the gene therapy of gliomas.

AIMTo fabricate multi-wall carbon nanotube (MWNT) modified electrode and study the electrochemical behaviors of diethylstilbestrol at the MWNT-modified electrode.

METHODSCyclic voltammetry and linear sweep voltammetry.

RESULTSThe oxidation peak current of diethylstilbestrol increased remarkably and the peak potential shifted negatively at the MWNT-dihexadecyl hydrogen phosphate (DHP) modified glassy carbon electrode (GCE), in contrast to that at the bare GC electrode and DHP-modified GC electrode. The oxidation peak current is linear with the concentration of diethylstilbestrol over the range from 1 x 10(-8) to 2 x 10(-6) mol.L-1. The detection limit was 2.5 x 10(-9) mol.L-1. The relative standard deviation (n = 10) was 2.9% for 1 x 10(-6) mol.L-1 diethylstilbestrol.

CONCLUSIONThe MWNT-DHP modified GCE exhibits catalytic activity to the oxidation of diethylstilbestrol.

Carbon ; chemistry ; Diethylstilbestrol ; analysis ; chemistry ; Electrochemistry ; Electrodes ; Hydrogen-Ion Concentration ; Nanotechnology

Objective:To investigate the genetic etiology of a child with epilepsy accompanied by motor retardation.Methods:A patient with epilepsy and motor retardation in Henan Provincial People′s Hospital in January 2020 and his parents′ peripheral blood 2 mL were collected.G-banded karyotyping and array-based comparative genomic hybridization (aCGH) were used to analyze the duplication / deletion of chromosome segments in child and her pa-rents.Results:The karyotype of the patient revealed 46, XX, and add(19)(p13.3→qter), whereas aCGH detected a 9.50 Mb duplication at 19q13.33q13.43[arr(hg19)(49593920_59092570)×3]. This region contains 471 genes.No abnormality was discovered in the karyotyping and aCGH analysis of the patient′s parents.The phenotypes of the patient conformed to the previously reported clinical characteristics of 19q13.3 duplication.Conclusions:The de novo 19q13.3 duplication is the cause of epilepsy and motor development retardation for the patient.Combined with aCGH, the traditional G banding is valuable to diagnose the patient with developmental delay.

Objective To investigate the clinical value of non-invasive prenatal diagnosis using cell free fetal DNA(cff-DNA)in maternal blood.Methods From Sep.2010 to Mar.2012,103 pregnant women who came to Henan Province People's Hospital in the first trimestcr for prenatal diagnosis of scx-linked inherited diseases were included in the first trimester group.From Oct.2010 to Jan.2012,205 pregnant women undergoing amniotic fluid sampling for fetal karyotype analysis in the same hospital were included in the second trimester group.Real time quantitative PCR and fluorescent PCR were used to detect sex determining region of Y chromosome gene(SRY)and amelogenin gene(AML)on cff-DNA of the first trimester group.Moreover,12 Y chromosome STR loci analysis were performed for 33 male fetuses and their fathers.Massively Parallel Signature Sequencing(MPSS)was used for aneuploidy analysis in cff-DNA of the second trimester group.Results(1)In the first trimester group,there were 53 SRY positive and 50 SRY negative.Compared with the results of cff-DNA of chorionic villus samples,there was one SRY false positive and one false negative results,with a sensitivity of 98％ and specificity of 98％.For the AML gene test,there were two PCR products of male fetuses:102 bp fragment originating from X chromosome(AML X)and 108 bp fragment from Y chromosome(AML Y);but only AML X was found in products from female fetuses.In the first trimester group,102 bp and 108 bp fragments were detected in 52 cases,and only 102 bp fragment was found in the other cases.Compared to AML results from chorionic villus samples,there were 2 false negative results,with a sensitivity of 96％ and specificity of 100％.(2)For cff-DNA with plasma SRY over 30 copy/ml,Y STR loci were analyzed on cff-DNA of 33 fetuses and their fathers.The Y STR loci less then 200 bp were successfully detected,while Y STR loci with PCR products between 200-300 bp showed low signal or could not be amplicated;and no PCR products more than 300 bp were detected from cff-DNA.Comparing the detected Y STR loci of cff-DNA to the fathers,32 fetuses were concordant with their fathers'.Exogenous contamination was found in the rest one sample.(3)In the second trimester group,6 fetuses with abnormal karyotype(two trisomy 21,three trisomy 18 and one 45,XO)were detected by cff-DNA and were proved by karyotype analysis.Moreover,the MPSS results of cff-DNA revealed one 45,Y and one trisomy 16 whose karyotype analysis showed normal results.And in one case,MPSS suggested less chrX or chrY,that was proved to be 47,XYY by karyotype analysis.Conclusions(1)Cff-DNA in maternal blood can be used to determine fetal gender in early prenancy with considerable sensitivity and specificity.But the trace cff-DNA and the high maternal DNA background might have impact on the result.(2)Analysis of cff-DNA in maternal blood of the second trimester women showed that MPSS could be used for prenatal screening of trisomy 21 and trisomy 18.However,further research should be done for other chromosomes aneuploidy detection.

It is always very difficult to process the bioelectrical signals, sampling, amplifying and quantifying because of the different sources it comes from and the mechanism it depends on. This paper mainly introduces a method of measuring the peak value of bioelectrical signals, expatiates on the structure, testing result, analysis for error and measure taken for improvement of the circuit designed by this method. At last, the paper also points out that the method is wonderful, simple, easy to carry out, and can be used in many medical systems.
Algorithms ; Brain ; physiology ; Electric Stimulation ; Electrophysiology ; instrumentation ; methods ; Equipment Design ; Microcomputers ; Neurons ; physiology ; Sensitivity and Specificity ; Signal Processing, Computer-Assisted ; Software

This paper mainly introduces the principles of a type of circuit for measuring frequency & duty cycle of low-frequency signals, and its applications in neuron-threshold stimulator. The circuit has the advantages of simple structure and accurate measurement.
Brain ; physiology ; Electrophysiology ; Equipment Design ; Humans ; Neurons ; physiology ; Physical Stimulation ; instrumentation ; Signal Processing, Computer-Assisted ; instrumentation

Objective - To analyze the effect of using vibration tools on the prevalence of work related musculoskeletal disorders （ ） Methods ， - WMSDs in automobile factory workers. By judgment sampling method front line workers with more than one year of working experience in an automobile factory were selected as the research subjects. Musculoskeletal Disorders Questionnaire was used for investigation. The workers were divided into the control group and the vibration tool group. The propensity score ∶ ， matching method was used to balance the confounding factors of the two groups of workers by 1 1 and 568 people were Results included in each group. The prevalence of WMSDs was compared between the two groups after matching. After ， ， ， ， ， ， matching the prevalence of WMSDs in the shoulder elbow hand/wrist upper back waist hip/buttock and knee of workers in ， （ P ） the vibration tool group was higher than that in the control group and the differences were statistically significant all <0.05 .， The prevalence of WMSDs in different body parts of workers in the vibration tool group ranking from high to low was waist ， ， ， ， ， ， ， ， ， ， neck shoulder hand/wrist upper back knee ankle/foot elbow and hip/buttock with the rate of 74.3% 61.3% 54.2% ， ， ， ， ， （P ） Conclusions 54.0% 50.9% 39.4% 35.2% 31.0% and 27.1% respectively <0.01 . The use of vibration tools can ， ， ， ， ， increase the risk of WMSDs in shoulder elbow hand/wrist upper back waist hip/buttock and knee of automobile factory workers. Corresponding measures should be taken to reduce vibration intensity and reduce contact time to protect workers'

OBJECTIVETo investigate the efficiency of multiplex ligation-dependent probe amplification (MLPA) combined with short tandem repeat (STR) linkage analysis for the prenatal diagnosis for Duchenne muscular dystrophy (DMD).

METHODSGender of the fetus was first determined by the presence of Y chromosome sex-determining gene (SRY). Subsequently, combined MLPA and STR linkage analysis were applied for the probands, pregnant women and fetuses in 45 affected families.

RESULTSAmong the 45 families, 31 SRY-positive fetuses were identified, among whom six were diagnosed with DMD. For 14 SRY-negative fetuses, four were diagnosed as carriers. The remainders were normal.

CONCLUSIONMLPA can detect mutations in the exons of dystrophin gene, whilst STR linkage analysis can determine whether the fetus has inherited the maternal X chromosome bearing the mutant gene. As the result, the method can detect affected fetuses in which no exonic mutations are detected with MLPA. By combining the two methods, the diagnostic rate for DMD can be greatly improved.

Dystrophin ; genetics ; Exons ; Female ; Genetic Linkage ; Heterozygote ; Humans ; Male ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne ; diagnosis ; genetics ; Mutation ; Pregnancy ; Prenatal Diagnosis

?AlM: To compare the clinical effect between small-incision cataract surgery and phacoemulsification.
?METHODS: Totally 93 patients ( 124 eyes ) with age-related cataract who received treatment in Mar 2010 and Feb 2013 were dicided into 2 groups randomly. Forty-two patients ( 59 eyes ) in group small - incision cataract surgery ( SlCS ) were treated by SlCS, while other 51 patients ( 65 eyes ) in group Phaco were treated by phacoemulsification. And then, postoperative visual acuity, corneal astigmatism, surgically induced astigmatism ( SlA ) and intraoperative and postoperative complications were contrasted between groups.
?RESULTS: After 1d and 1wk of postoperation, there were 38 eyes ( 64. 4%) and 41 eyes ( 69. 5%) having a better visual acuity of 0. 5 in the SlCS group, while there were 29 eyes (44. 6%) and 32 eyes (49. 2%) in the Phaco group. The vision of SlCS group was better than that of Phcao group (χ2 = 4. 877, 5. 242, P < 0. 05 ). On postoperative 1 and 3mo, with acuity of 0. 5 or better, eye numbers showed no statistically significant differences between two groups (χ2 = 0. 005, 0. 085, P>0. 05). The average corneal astigmatism used analysis of repeatedly measuring designing variance: Comparing the corneal astigmatism in intra - groups at different times, it was statistically significant (F=25. 624, P<0. 05), and had a tendency to decrease with time. However, there was no statistical significance for corneal astigmatism between groups (F=0. 986, P>0. 05). The coneal astigmatism of each group was higher at 1wk after the surgery than that of preoperation, and the contrast had statistical sigenficence (t=2. 906, 2. 427, P<0. 05). The Phaco group with SlA was lower than the SlCS group at 1wk and 1mo after the surgery (t=-4. 628, 2. 770, P<005). lt had no statistical significance in SlA by comparing with the two groups at 3mo after the surgery (t=0. 754, P>0. 05). There were statistical differences in SlA at different time both by intra-group comparison and group comparison ( F=26. 37, P<0. 05, F = 14. 29, P<0. 05). The comparison of posterior capsule rupture, the postoperative corneal edema and anterior chamber pigment membrane reaction in two groups showed no statistical significance.
?CONCLUSlON: Our research shows that small-incision cataract surgery and phacoemulsification had similar effect in the treatment of cataract. Phacoemulsification is not the only surgery option for the best treatment effect. Small- incision cataract surgery can be popularized in basic- level hospitals, achieving the effect similar to phacoemulsification.