How Job Stress and Psychological Adaptation Predicting Interpersonal Needs Among Female Migrant Manufacturing Workers in China: A Structural Equation Model

Background@#This study aimed to explore relationships between job stress and psychological adaptation and how they related to interpersonal needs through mood states among female migrant manufacturing workers. @*Methods@#A cross-sectional survey was conducted in 16 factories in Shenzhen, China. Sociodemographic, job stress, psychological adaptation and other psychological information of was collected. Structural equation modeling was performed to delineate the internal relationship between variables. @*Results@#The hypothetical structural equation model exhibited acceptable model fit among female migrant manufacturing workers (χ2 = 11.635, df = 2, χ2/df = 5.82, p = 0.003, RMSEA = 0.090, CFI = 0.972, SRMR = 0.020). Job stress was directly associated with mood states and interpersonal needs; Psychological adaptation was directly associated with mood states and indirectly associated with interpersonal needs; Bootstrapping tests demonstrated mediation effect of mood states in the way from psychological adaptation to interpersonal needs. @*Conclusion@#Female migrant manufacturing workers who suffered stress from job and the process of psychological adaptation may have worse mood states and workers with worse mood states are more likely to develop unmet interpersonal needs, a proximal factor of suicidal ideation.

Clinical Analysis of Bloodstream Infection after Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

OBJECTIVE@#To analyze the clinical characteristics of bloodstream infection (BSI) in patients treated by hematopoietic stem cell transplantation (HSCT).@*METHODS@#The clinical characteristics, distribution of pathogenic bacteria causing BSI and drug sensitivity of 910 patients treated by HSCT in our department from January 2013 to June 2020 were retrospectively analyzed.@*RESULTS@#Among 910 HSCT patients, 111 patients were diagnosed as BSI within 100 days after transplantation, and 98 patients showed BSI during the period of agranulocytosis. Multivariate analysis showed that the usage of anti-thymocyte globulin (ATG), long duration of agranulocytosis and low infusion volume of mononuclear cell (MNC) were the independent risk factors affecting BSI after HSCT. Among 121 pathogenic bacteria isolated, 76 Gram-negative (G-) bacteria (62.8%), 40 Gram-positive (G+) bacteria (33.0%), and 5 fungi (4.1%) were detected out. The top three pathogens were Escherichia coli, Staphylococcus epidermidis and Pseudomonas aeruginosa. The drug-resistance rates of Escherichia coli and Klebsiella pneumoniae to carbapenems was 14.3% and 7.7%, respectively, and Pseudomonas aeruginosa was 66.7%. The susceptibility of G+ bacteria to vancomycin, linezolid and teicoplanin was 97.5%, 100% and 100%, respectively. The crude mortality rate of the patients with BSI at 100 days after HSCT was significantly higher than that of patients without BSI (P<0.001).@*CONCLUSION@#The usage of ATG, long duration of agranulocytosis and low infusion volume of MNC are independent risk factors for BSI after HSCT. The pathogens after HSCT are mainly G- bacteria. Pseudomonas aeruginosa is highly resistant to carbapenems. Key words　　;

The clinical application of mobile internet remote guidance platform for vestibular rehabilitation / 中华耳鼻咽喉头颈外科杂志

Objective: To guide the patients with vertigo who are suitable for vestibular rehabilitation therapy (VRT), and to evaluate the curative effect through a remote guidance platform based on mobile internet. Methods: Adult outpatients, who were diagnosed as vestibular disorders and required VRT, were selected and conducted baseline evaluation and formulated vestibular rehabilitation plan according to their symptoms, diagnosis and vestibular function examination results. These patients downloaded and installed the mobile internet remote guidance platform app for VRT, and then registered and uploaded medical records. According to the VRT plan formulated by clinicians for patients, the platform launched corresponding exercise guidance videos to guide them to complete 4-week VRT exercise at home. Before and after VRT, the patients were scored with Visual Analogue Scale (VAS), Activities-specific Balance Confidence (ABC), Dizziness Handicap Inventory (DHI) and Self-rating Anxiety Scale (SAS). The rehabilitation effects were statistically analyzed by SigmaStat 4.0 software. Results: From October 2019 to October 2021, 233 patients with vertigo completed the registration of vestibular rehabilitation guidance platform, of whom 187 patients insisted on 4-week rehabilitation training and completed the scale evaluation. Among 187 patients, 65 were male and 122 were female; Age was (49.8±16.0) years; The medical history ranged from one to 192 months, with a median of eight months. Compared with that before rehabilitation exercise, the subjective feeling of vertigo in 170 patients was improved, and the overall effective rate was 90.9% (170/187). The subjective symptoms of vertigo were basically improved after rehabilitation training in patients with unilateral vestibular dysfunction, vestibular neuritis, sudden deafness with vertigo, Hunt syndrome and acoustic neuroma. There were significant differences in ABC, DHI and SAS scores before and after VRT (P<0.05). Of those patients with Meniere's disease in the intermittent period and the patients with Meniere's disease who underwent surgical treatment, more than 90% of their subjective symptoms of vertigo or dizziness improved after VRT, and there were significant differences in the scores of ABC, DHI and SAS before and after VRT exercise (P<0.05). In patients with vestibular migraine, 36.7% (11/30) had no improvement or even aggravation of subjective symptoms of vertigo after VRT, however, the DHI score after rehabilitation exercise was lower than that before exercise, and the difference was statistically significant (P<0.05). In patients with bilateral vestibular dysfunction, although most (6/8) subjective symptom scores were improved compared with those before exercise, there was no significant difference in ABC, DHI and SAS scores before and after rehabilitation (P>0.05). Conclusion: VRT with the help of vestibular rehabilitation mobile internet remote guidance platform can effectively improve the subjective symptoms of vertigo, balance ability and anxiety in patients with unilateral vestibular lesions.

Clinical and Bacteriological Analysis of Bacterial Bloodstream Infections in Patients with Acute Leukemial / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical characteristics, etiology and drug susceptibility of bacterial bloodstream infections in acute leukemia(AL) patients.@*METHODS@#Clinical data, etiology and drug susceptibility of acute leukemia patients with bacterial bloodstream infections from April 2009 to April 2018 were retrospectively analyzed.@*RESULTS@#A total of 376 strains were isolated, 76.9% was Gram-negative bacterial and 23.1% was Gram-positive bacteria. Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae were listed as the top three of Gram-negative bacteria. The susceptibility of Escherichia coli to the tigacycline, imipenem and meropenem was 100.0%, 98.2% and 98.1%, respectively. The susceptibility of Klebsiella pneumoniae to the tigacycline, imipenem and meropenem were 100.0%, 98.3% and 94.4%, respectively. The adjustment rate for initial use of carbopenems was 3.8%, while the adjustment rate for initial use of noncarbopenems was 74.3% in patients with main Gram-negative bacterial blood stream infection. The susceptibility of Gram-positive bacteria to glycopeptide antibiotics, linezolid and tigacycline was 100.0%.@*CONCLUSION@#Gram-negative bacteria is the majority type of bacteria in AL patients with bacteria blood stream infections. The susceptibility of Gram-negative bacteria to the carbapenems is high, and the treatment adjustment rate is obviously low. The glycopeptide, linezolid and tigacycline are effective for Gram-positive bacteria infections..

Effect of rotary cell culture system-simulated microgravity environment on the expression of lncRNA in mouse fibroblasts / 解放军医学杂志

Objective To investigate the effects of simulated microgravity by rotary cell culture system (RCCS) on expression profiles of long non-coding RNA (lncRNA) in mouse fibroblasts L929 cell line.Methods L929 cells were cultured in vitro and randomly divided into simulated microgravity (SMG) group and normal gravity (NG) group.Each group had three samples,the rotator axis of SMG group was paralleled to the ground rotation,while the rotator axis of NG group was vertical to the ground rotation,and the speed of rotation was consistent for the two groups.The samples of two groups were collected on 7th day of culture and the total RNAs were extracted,labeled and hybridized in sequence.The lncRNA and mRNA were detected by Agilent Mouse lncRNA Chips respectively.Differentially expressed lncRNA were identified and then validated by RT-qPCR.GO and Pathway analysis were applied to determine the functional distribution of these target genes.The integration predictions of the lncRNA and mRNA co-expression had been proposed to refine the functional lncRNA-mRNA relationships.Results There were 238 differentially expressed lncRNAs including 134 lncRNAs up-regulated and 104 lncRNAs down-regulated,and 237 differentially expressed mRNAs including 53 mRNAs up-regulated and 184 mRNAs down-regulated significantly in mouse fibroblasts L929 cell line under simulated microgravity by RCCS.The RT-qPCR showed a high concordance with chip microarray results in 4 differentially expressed lncRNA.GO analysis showed that the differentially expressed lncRNAs were related to the biological processes such as negative regulation of megakaryocyte differentiation and negative regulation of wound healing.Pathway analysis showed that these target genes were related to the signal pathways of systemic lupus erythematosus and TGF-β.The lncRNA-mRNA co-expression networks were also established.Conclusion The simulated microgravity by RCCS could significantly affect the expression profiles of lncRNA and mRNA in mouse fibroblasts L929.The lncRNA target genes prediction and functional enrichment analysis based on gene chip technology may provide the theoretical basis for illustrating the mechanism and management of weightlessness stress injury.

Study on Accurately Controlling Discharge Energy Method Used in External Defibrillator / 中国医疗器械杂志

This paper introduces a new method which controls discharge energy accurately. It is achieved by calculating target voltage based on transthoracic impedance and accurately controlling charging voltage and discharge pulse width. A new defibrillator is designed and programmed using this method. The test results show that this method is valid and applicable to all kinds of external defibrillators.

Preliminary study of low-dose CT coronary angiography by using low concentration isotonic contrast agent / 中华放射学杂志

Objective To compare the iodine intake,image quality and radiation dose of dual-source CT coronary artery angiography between the low concentration isotonic contrast agent with iterative reconstruction technique of sinogram affirmed iterative rEconstruction (SAFIRE) and common concentration contrast agent with filtered back projection (FBP).Methods One hundred patients [58 men,42 women; age:(62± 11)y] were enrolled in this study.Fifty consecutive patients (Group A) were scanned with FBP and common concentration contrast agent and another 50 consecutive patients (Group B) were scanned with low concentration isotonic contrast agent and iterative reconstruction technique.The image quality were assessed by two experienced radiologists with a double blinded fashion in a five score scale.The attenuation of coronary artery,scan time,imaging noise,signal-to-noise ratio (SNR),contrast-to-noise ratio (CNR),radiation dose and iodine intake between these two groups were compared using two sample t test.The observer agreement for image quality was calculated using Kappa statistics.Results All examinations were completed successfully.There was no statistical difference (P＞0.05) between two groups in clinical characteristics and scan parameters.There was no statistical difference for the mean subjective image quality of group A (4.4±0.7) and group B (4.3±0.8) (t=0.924,P＞0.05).The Kappa value between two radiologists was 0.887 (P＜0.01)on image quality.The mean attenuation of coronary artery segments was higher than 300 HU,especially in group A (386.1±51.5) HU,which was slightly higher than the group B (384.1±77.1) HU,but there was no statistical difference (t=0.157,P＞0.05).The SNR and CNR in two groups did not have significant differences (P＞0.05).The total iodine and iodine injection rates were 21.0 g,17.5 g/s in Group A and 16.2 g,13.5 g/s in Group B,respectively.The iodine intake was decreased by 22.9％ in Group B.The effective radiation dose in Group B (1.09±0.19) mSv was significantly (t=20.260,P＜0.01) lower than that in Group A (2.85±0.59) mSv,with the radiation dose reduced by 61.8％.Conclusion Low concentration isotonic contrast agent and iterative reconstruction technique can significantly reduce the radiation dose and iodine intake in CT coronary artery angiography and achieve the same image quality using common concentration contrast agent with FBP.

Mechanism of GLI3 gene transcription regulation in idiopathic congenital talipes equinovarus / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the mechanism of transcription regulation of GLI3 gene in idiopathic congenital talipes equinovarus.</p><p><b>METHODS</b>pGL3-Gli3 luciferase report vectors were constructed, and the activity of Gli3 promoter was explored. A P-Match software was used to analyze the sequence upstream of the transcription start site of rat Gli3 gene, which was subsequently verified with chromatin immunoprecipitation assay (CHIP) and electrophoretic mobility shift assay (EMSA). Expression of the Gli3 gene was analyzed in L6 cells transfected with Hoxd13 small interference RNA(siRNA) and Hoxd13 expression vectors.</p><p><b>RESULTS</b>The 5' region of rat Gli3 gene contains two potential binding sites for the Hoxd13 protein. CHIP and EMSA assays both confirmed that Hoxd13 can directly bind with site 2. As shown in L6 cells, expression of Gli3 may be enhanced with silencing of Hoxd13, whilst exogenous expression of Hoxd13 can down-regulate transcription of Gli3.</p><p><b>CONCLUSION</b>Hoxd13 can directly regulate the expression of Gli3 gene through a Hoxd13 binding site in the limb of rat embryo.</p>

Possible role of GLI3 gene in the pathogenesis of idiopathic congenital talipes equinovarus / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between GLI3 gene and pathogenesis of idiopathic congenital talipes equinovarus (ICTEV).</p><p><b>METHOD</b>Potential mutations in the coding region of GLI3 were detected among 84 patients with ICTEV by denaturing gradient electrophoresis. Expression of GLI3 in the ICTEV patients' disease tissues was assessed by reverse transcription PCR. Following generation of rat model for ICTEV, mRNA and protein levels of GLI3 were evaluated by real-time PCR and immunohistochemistry and Western blotting.</p><p><b>RESULTS</b>No mutation was found in exons 1 - 8 and 13 of GLI3 gene among the 84 ICTEV patients. No expression of GLI3 gene was detected in the flexor hallucis longus of ICTEV patients or normal controls. Expression of Gli3, in terms of both mRNA and protein, was stronger in the hindlimb of ICTEV rat embryos compared with normal controls.</p><p><b>CONCLUSION</b>Mutation in the coding region of GLI3 may not be responsible for the occurrence of ICTEV. However, there may still be connection between abnormal expression of the gene and pathogenesis of ICTEV.</p>

Expression of COL9A1 gene and its polymorphism in children with idiopathic congenital talipes equinovarus / 中国当代儿科杂志

<p><b>OBJECTIVE</b>COL9A1 gene is located in the susceptibility region of idiopathic congenital talipes equinovarus (ICTEV) (6q12-13). This study aimed to investigate the expression of the COL9A1 gene and the distribution of single nucleotide polymorphism (SNP) of COL9A1 gene in patients with ICTEV and normal controls.</p><p><b>METHODS</b>Immunohistochemistry was used to detect the expression of COL9A1 in 25 children with ICTEV and 5 normal controls. The frequencies of genotypes and allele of two SNPs in COL9A1 gene rs35470562 and rs1135056 were investigated by PCR-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing in 118 patients with ICTEV and 100 normal controls.</p><p><b>RESULTS</b>The COL9A1 protein expression was significantly higher in 22 (88%) out of 25 children with ICTEV than normal controls. There were significant differences in the frequencies of genotypes and allele of rs1135056 in COL9A1 gene between the ICTEV and the control groups: the G allele frequency was higher, the frequency of AA genotype was lower, and the frequencies of AG and GG genotypes were higher in ICTEV patients than those in healthy controls (P<0.05).</p><p><b>CONCLUSIONS</b>COL9A1 protein is highly expressed in patients with ICTEV and rs1135056, which is located in the coding region of COL9A1 gene, may be associated with the pathogenesis of ICTEV.</p>

Dual regulation effect of somatostatin on immunity in patients with severe sepsis caused by abdominal diseases / 中华外科杂志

<p><b>OBJECTIVE</b>to investigate the effect of somatostatin on inflammatory immune disorders and prognosis in patients with severe sepsis caused by abdominal diseases.</p><p><b>METHODS</b>fifty-three patients with severe abdominal sepsis (age > 18 years, APACHE-II score > 15) from June 2005 to June 2009 were randomly divided into Somatostatin group (n = 23) and SSC Group (n = 30). Fifteen healthy volunteers of the same age range were chosen as Control group. The SSC group was treated with classical SSC therapy, and the Somatostatin Group was treated with the same regime plus 14-peptide somatostatin continuous infusion at the dose of 6 mg/24 h for 7 days. The serum levels of interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α) were determined by using ELISA. CD(4)(+), CD(8)(+) T cell subsets were determined by fluorescence activated cell sorter(FACS) and CD(4)(+)/CD(8)(+) was calculated. APACHE-II score was observed on admission (d1) and day 3, 7 and 14 after treatment. Morality rates in 28 days in two groups were recorded.</p><p><b>RESULTS</b>compared with Control group, IL-10 and TNF-α levels were significantly elevated in patients with severe abdominal sepsis (P < 0.05), while CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) decreased significantly (P < 0.05). Compared with the Somatostatin group CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) on d7 and d14 in SSC Group were significantly increased (P < 0.05), while IL-10 and TNF-α decreased significantly(P < 0.05). APACHE-II scores on d3, d7, d14 of Somatostatin group were significantly lower than those of SSC group, and 28 d mortality rate also declined.</p><p><b>CONCLUSIONS</b>in patients with severe abdominal sepsis, systemic inflammatory response and immune suppression exist simultaneously. Somatostatin has a dual immunomodulatory activity in these patients.</p>

Molecular and prenatal diagnosis of a pedigree with spinocerebellar ataxia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify the type of a pedigree with spinocerebellar ataxia, and carry out asymptomatic carrier detection and prenatal diagnosis.</p><p><b>METHODS</b>The blood samples of two patients in the spinocerebellar ataxia pedigree were collected. Based on the clinical characteristics of the pedigree and the disease incidence in China, the regions containing the CAG repeat of the SCA1, SCA2 and SCA3/MJD genes were amplified by polymerase chain reaction (PCR). The numbers of CAG repeats in the normal and abnormal allele fragments were identified by using agarose gel electrophoresis and DNA sequencing. We further carried out tests on the children of the patients and fetus to identify the presence of the abnormal allele.</p><p><b>RESULTS</b>The numbers of CAG repeat in the SCA1 and SCA2 genes were in the normal range. The CAG repeat number in one allele of SCA3/MJD gene was in the normal range, while that in the other allele was in the abnormal range. One of the children of the patients and the fetus carried the abnormal allele.</p><p><b>CONCLUSION</b>It was confirmed that the pedigree was SCA3/MJD by gene diagnosis. One of the children of the patients was asymptomatic carrier and the fetus also carried the abnormal allele.</p>

Gene diagnosis for spinal muscular atrophy and its application study / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish an effective testing system for gene diagnosis, carrier detection and prenatal diagnosis for spinal muscular atrophy (SMA).</p><p><b>METHODS</b>Twenty-six patients with SMA were directly tested with PCR-RFLP for exon 7 deletion in the SMN1 gene. Carrier detection was carried out with multi-PCR-DHPLC. Amniotic fluid was taken at the middle stage of gestation from pregnant women who had given birth to affected children.</p><p><b>RESULTS</b>Twenty-five out of 26 patients were diagnosed as having SMN1 gene deletion. Fifty-two of their parents were found to be carriers of exon 7 deletion. Eight of 20 fetuses were diagnosed as having SMN1 gene deletion by PCR-RFLP.</p><p><b>CONCLUSION</b>PCR-RFLP and multi-PCR-DHPLC techniques can provide rapid diagnosis for exon 7 deletion detection and carrier detection. PCR-RFLP may also be adapted for prenatal gene diagnosis of exon 7 deletion in SMN1 gene.</p>

Non-invasive prenatal genetic diagnosis using multiple displacement amplification / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of multiple displacement amplification (MDA) to apply in the non-invasive prenatal genetic diagnosis of Duchenne muscular dystrophy (DMD).</p><p><b>METHODS</b>Maternal blood was obtained from 20 pregnant women at 7 to 25 weeks of gestation. After the discontinuous density gradient centrifugation with Percoll, the fetal nucleated red blood cells (NRBCs) were stained with Kleihauer test. All positive NRBCs were collected by micromanipulator and then performed with MDA. Sex and short tandern repeat (STR) analysis were determind from a small aliquot of the reaction. The origin of NRBCs was verified and prenatal diagnosis of DMD was made at the same time.</p><p><b>RESULTS</b>The product length of MDA was >15 kb, while primer extension preamplification (PEP) is only about 1 kb. We completed non-invasive prenatal genetic diagnosis of 6 fetus at high risk of DMD using MDA. The results were all coincident with amniotic fluid control.</p><p><b>CONCLUSION</b>The MDA method which provides a highly uniform representation across the genome, representing the entire genome with minimal amplification bias, shows good application prospects.</p>

Application study on inversion diagnosis of F8 gene in hemophilia A / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish an effective method of genetic diagnosis on hemophilia A (HA) by detecting the inversion mutation in intron 22 of F8 gene.</p><p><b>METHODS</b>Intron 22 inversion mutation in F8 gene was detected by using long distance-polymerase chain reaction (LD-PCR) and inversion-PCR (I-PCR) in 31 HA patients. The mothers of HA patients with intron 22 inversion mutation were selected to carrier diagnosis and amniotic fluid of the pregnant women with inversion mutation was collected at intermediate stage of gestation, and used to prenatal genetic diagnosis.</p><p><b>RESULTS</b>Seven patients showed F8 gene inversion mutation in thirty-one patients. Three in four mothers of HA patients with intron 22 inversion mutation were diagnosed as carriers. The prenatal diagnosis result indicated that the fetus conceived in the HA-carrier woman was normal individual.</p><p><b>CONCLUSION</b>The detection of intron 22 inversion mutation by LD-PCR and I-PCR is time-saving, and can be used in prenatal diagnosis on HA.</p>

Proteomic analysis of the ankle joint bone, ankle joint tissue and spinal cord of clubfoot-like deformity in rat fetuses / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the etiology of idiopathic talipes equinovarus (ITEV) in all-trans retinoic acid (ATRA) induced clubfoot-like deformity in rat fetuses with two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS).</p><p><b>METHODS</b>Clubfoot-like deformity model in rat fetuses was induced with ATRA (135 mg/kg) in gestation day (GD10) pregnant Wistar rats. 2-DE was applied to separate the total proteins of ankle joint tissue, ankle joint bone and spinal cord of the animal models. The Coomassie Brilliant Blue staining gels were analyzed by 2-DE software PDQuest 7.1.0. Selected differential protein spots were identified with peptide mass fingerprinting based on matrix-assisted laser adsorption/ionization time-of-flight mass spectrometry and database searching. xiap, tnnt1 and col2 alpha 1, three genes of the differential proteins, were identified furthermore. Apoptosis study was made in terminal deoxynucleotidyl transferase nick end labeling.</p><p><b>RESULTS</b>There were many differential expressed proteins in the clubfoot-like deformity model. Out of the differentially expressed proteins,16 protein spots were identified to be differentially expressed in the clubfoot-like deformity model with MS. Three of the 16 protein spots, xiap, tnnt1 and col2 alpha 1 were confirmed to be significantly down-regulated by the RT-PCR, and Xiap was further confirmed to be significantly down-regulated with immunohistochemistry. Another randomly selected gene, ngfr, did not express differently in ATRA-induced clubfoot-like deformity in rat fetuses. The rates of the apoptosis in the spinal, bone of the clubfoot-like deformity fetuses was 5.4 and 10 times of those of the normal fetuses respectively.</p><p><b>CONCLUSION</b>The results suggest that there are certain differently expressed proteins in ankle joint tissue, ankle joint bone and spinal cord of the ATRA-induced clubfoot-like deformity in rat fetuses, and Xiap, sTnT, and Col2 alpha 1 show a significant correlation with ITEV. Ngfr is not correlation with ITEV. Apoptosis plays a key role in the development of ITEV and related to the decreased expression of the Xiap.</p>

Detection of new mutations in the dystrophin gene by denaturing high-performance liquid chromatography / 中华儿科杂志

<p><b>OBJECTIVE</b>Duchenne muscular dystrophy (DMD) is an X-linked recessive disease caused by dystrophin gene mutations; 55%-65% of these pathogenic mutations are large deletion and duplication mutations that can be detected by multiplexed polymerase chain reaction. However, finding the remaining micro-mutations (substitutions, deletions or insertions of one or several nucleotides) cannot be achieved in this way. The aim of the present study was to detect mutations of the dystrophin gene in individuals with Duchenne muscular dystrophy (DMD) by denaturing high-performance liquid chromatography (DHPLC) and to establish a rapid and sensitive screening platform for micro-mutations leading to DMD.</p><p><b>METHODS</b>Twenty patients negative for large deletions in the dystrophin gene by multiplex PCR were selected for further screening by DHPLC and 20 normal male without DMD family history as the control cohort. Dystrophin exons and their flanking sequences were individually amplified by genomic PCR and the amplicons showing abnormal DHPLC profile were directly sequenced to identify the position and the type of the mutations.</p><p><b>RESULTS</b>After screening 68 exons covering the two deletion hotspots and 3'UTR region, four pathogenic mutations, including c.6808_6811del TTAA, c.4959_4960insA, c.8656C > T and c.8608C > T, were found in four DMD patients. Moreover, c.6808_6811del TTAA, c.4959_4960ins and c.8656C > T have not been reported previously. The first two frameshift mutations were predicted to produce premature stop codons, p.Leu2270MetfsX9 and p.Ser1654LysfsX5, respectively. The remaining two were nonsense mutations, leading to p.R2886X and p.R2870X, respectively.</p><p><b>CONCLUSION</b>Three novel and one recurrent dystrophin mutations have been identified in Chinese DMD patients. This study has demonstrated that DHPLC is an effective screening method for micro-mutation associated with DMD.</p>

Association and mutation analysis of GLI3 gene in idiopathic congenital talipes equinovarus / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the association and mutation of GLI3 gene in idiopathic congenital talipes equinovarus(ICTEV).</p><p><b>METHODS</b>(1) Genotype of 2 single nucleotide polymorphism (SNP) in 84 idiopathic congenital talipes equinovarus nuclear pedigree were analyzed by restriction fragment length polymorphism. Association analysis was directed between single SNP locus and ICTEV through ETDT software, respectively.(2) Mutation sites in exon 9,10,11,12 of GLI3 gene were detected in 103 patients with ICTEV by denaturing gradient gel electrophoresis technique.</p><p><b>RESULTS</b>rs929387ls located in exon 14 of GLI3 gene have transmission disequilibrium in 84 nuclear pedigrees (P<0.05), and rs846266 located in exon 4 have no transmission disequilibrium (P>0.05). A synonymous mutation in exon 9 was detected in one patient and his mother.</p><p><b>CONCLUSION</b>There is an association between GLI3 gene and ICTEV, and exons 9,10,11,12 are not its mutation hot spots.</p>

Evaluation of Domestic Cryostorage Blood Bags / 대한수혈학회지

BACKGROUND: Recent studies have shown that the optimal temperature at which the cord blood and the peripheral blood stem cells need to be cryopreserved is below -160 degrees C in a nitrogen tank. This study evaluated a domestic cryostorage cord blood processing bag (CBP set) made from poly (ethylene-co-vinylacetate) (EVA) recently produced by the Medi-Rution Company in Korea. METHODS: The domestic cryostorage bags was evaluated using 100 bags from Medi-Rution, which were divided into 3 different groups and compared with 20 bags produced by the Thermogenesis company from the United States as a control. In the first group, the mononuclear cells separated from the human blood sample were divided equally into the cryosolution using DMSO, auto plasma and RPMI media. The blood was preserved in 20 domestic bags and 20 bags produced by the Thermogenesis company. In the second group, the cells were stored in the cryosolution with either auto plasma or fetal bovine serum. In the third group, 20 bags were frozen at -70 degrees C before being stored in the nitrogen tank whereas the other 20 bags were frozen in a controlled refrigerator before being stored in the nitrogen tank at -196 degrees C. The bags containing the peripheral blood cells were stored in the nitrogen tank at vapor phase for a period of one year and evaluated for the rate of breakage during thawing, recovery rates and viability of the cell after thawing. RESULTS: None of the bags evaluated in this study showed any signs of breakage or leakage of the products. All three groups tested showed similar results regardless of where the bags were made, the composition of the cryosolution or the early freezing conditions. CONCLUSION: The CBP set cryostorage bags recently produced by Medi-Rution can be used to preserve blood cells at extremely low temperatures for a one year period without any significant breakage.

Analysis of association between 5' HOXD gene and idiopathic congenital talipes equinovarus / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>Four single nucleotide polymorphisms (SNP) in HOXD10, HOXD12 and HOXD13 genes were chosen to investigate SNP and haplotypes distribution in idiopathic congenital talipes equinovarus nuclear pedigrees.</p><p><b>METHODS</b>Genotypes of 4 SNPs in 84 idiopathic congenital talipes equinovarus nuclear pedigrees were analyzed by restriction fragment length polymorphism and DNA sequencing. Analysis of association between SNP locus and idiopathic congenital talipes equinovarus was performed using ETDT software. Haplotypes and their frequencies in 84 nuclear pedigrees were established and analyzed by TRANSMIT software.</p><p><b>RESULTS</b>rs847151 polymorphism was not detected; the rs847154 located in 5' flanking sequence of HOXD12 gene and the rs13392701 located in exon 1 of HOXD13 gene were noted to have transmission disequilibrium in 84 nuclear pedigrees (P < 0.05).</p><p><b>CONCLUSION</b>rs847154 located in 5' flanking sequence of HOXD12 gene and rs13392701 located in exon 1 of HOXD13 gene are associated with idiopathic congenital talipes equinovarus; HOXD12 andHOXD13 are important susceptible genes of idiopathic congenital talipes equinovarus.</p>