Molecular cytogenetic study of a case with ring chromosome 15 / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a patient featuring developmental delay.@*METHODS@#The patient and her parents were subjected to G- and C-banded chromosomal karyotyping analysis. The proband was also analyzed by single nucleotide polymorphism microarray (SNP-array). The result was verified by using fluorescence quantitative PCR (qPCR).@*RESULTS@#The proband's karyotype was ascertained as 46,XX, r(15)(p11.2q26.3)[92]/45,XX,-15[9]/46,XX, dic r(15)(p11.2q26.3;p11.2q26.3)[4]. SNP-array revealed that she has carried a de novo deletion at 15q26.3 (98 957 555-102 429 040) spanning approximately 3.4 Mb, which encompassed the IGF1R gene. qPCR has confirmed haploinsufficiency of exons 3, 10 and 20 of the IGF1R gene. Both of her parents had a normal karyotype.@*CONCLUSION@#The abnormal phenotype of the proband may be attributed to the microdeletion at 15q26.3, in particular haploinsuffiency of the IGF1R gene and instability of the ring chromosome. Cytogenetic method combined with SNP-array and qPCR can efficiently delineate chromosomal aberrations and provide accurate information for clinical diagnosis and genetic counseling.

Genetic study of a child with developmental delay and mental retardation / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a child with developmental delay and mental retardation.@*METHODS@#Chromosomal karyotype of the child was analyzed by G-, C- and N-banding techniques. Her genome DNA was analyzed with single nucleotide polymorphisms array (SNP array). The result was validated by fluorescence quantitative polymerase chain reaction (PCR).@*RESULTS@#The karyotype of the child was ascertained as 46,XX,r(22)(p12q13). SNP array has revealed a deletion of approximately 1.4 Mb at 22q13.33 (49 802 963-51 197 766). The deletion has encompassed the SHANK3, a crucial gene for the development of nervous system. Fluorescence quantitative PCR has confirmed the deletion of exons 7, 19 and 22 of the SHANK3 gene.@*CONCLUSION@#The phenotype of the patient may be attributed to the microdeletion at 22q13.33. Cytogenetic methods combined with SNP array and fluorescence quantitative PCR can identify aberrant chromosomes and provide accurate information for the clinical diagnosis and genetic counseling.

Application of high-throughput whole genome sequencing and STR typing for the analysis of chorea villus tissue samples from spontaneous abortion / 中华医学遗传学杂志

OBJECTIVE@#To apply high-throughput whole genome sequencing (WGS) and short tandem repeat (STR) typing to detect aneuploidies, heteroploidies and copy number variations(CNVs) in spontaneous abortic tissues.@*METHODS@#Chorionic villus samples from 145 patients with spontaneous abortion were subjected to detection of aneuploidies, heteroploidies and copy number variations by WGS and STR typing.@*RESULTS@#All testing was successful and the rate of chromosomal abnormalities among the patients was 22.07%. Among these, there were 11 trisomies, 3 monosomies, 2 triploidies, 5 autosomal mosaicisms, 4 sex chromosomal mosaicisms, 7 structural abnormalities (including 1 mosaicism). In 89 cases, there were 130 CNVs of uncertain significance, 47 likely benign CNVs, and 2 loss of one copy of pathogenic AR gene. One sample contained 6 fragment duplications and deletions. Only 24 samples had no abnormal finding.@*CONCLUSION@#The most important reason for spontaneous abortions is embryonic chromosomal abnormality. Combined STR typing and WGS is both comprehensive and fast, and may become a major means for the detection of chorionic villi tissue from spontaneous abortions.

Application of high-throughput whole genome sequencing and STR typing for the analysis of chorea villus tissue samples from spontaneous abortion / 中华医学遗传学杂志

Objective@#To apply high-throughput whole genome sequencing (WGS) and short tandem repeat (STR) typing to detect aneuploidies, heteroploidies and copy number variations(CNVs) in spontaneous abortic tissues.@*Methods@#Chorionic villus samples from 145 patients with spontaneous abortion were subjected to detection of aneuploidies, heteroploidies and copy number variations by WGS and STR typing.@*Results@#All testing was successful and the rate of chromosomal abnormalities among the patients was 22.07%. Among these, there were 11 trisomies, 3 monosomies, 2 triploidies, 5 autosomal mosaicisms, 4 sex chromosomal mosaicisms, 7 structural abnormalities (including 1 mosaicism). In 89 cases, there were 130 CNVs of uncertain significance, 47 likely benign CNVs, and 2 loss of one copy of pathogenic AR gene. One sample contained 6 fragment duplications and deletions. Only 24 samples had no abnormal finding.@*Conclusion@#The most important reason for spontaneous abortions is embryonic chromosomal abnormality. Combined STR typing and WGS is both comprehensive and fast, and may become a major means for the detection of chorionic villi tissue from spontaneous abortions.

Diagnosis of a case with partial 9p trisomy by next generation sequencing / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic cause for a child featuring growth and mental retardation.@*METHODS@#Following conventional karyotyping analysis of the trio family, next generation sequencing (NGS) was carried out to explore the origin of the supernumerary marker chromosome. Fluorescence in situ hybridization (FISH) was used to confirm the result.@*RESULTS@#The karyotypes of both parents were normal, while the proband was found to be 47,XX,+mar. NGS showed that the supernumerary marker has originated from chromosome 9p13.1p24.3 with a size of 39.77 Mb. FISH has confirmed the above finding.@*CONCLUSION@#The 9p13.1-p24.3 trisomy probably underlies the abnormal phenotypes of the child. Cytogenetic analysis combined with NGS and FISH can provide accurate diagnosis for such disorders.

Application and Prospect of Automatic Control Technology in Extraction of Chinese Medicine / 世界科学技术-中医药现代化

In our country, the technology of extraction and separation of traditional Chinese medicine is backward relatively. The production process is extensive and the equipment level is low relatively. The manufacturing process is mainly based on unit operation and manual operation. The automatic control of the whole process has not been realized, which has seriously restricted the modernization of traditional Chinese medicine industry. If automatic control technology applied to traditional Chinese medicine in the extraction process, the process parameters of operation will be in the effective and strict monitoring and control, so as to improve the product quality and production efficiency, reduce costs, to achieve the green production. In this paper, the current situation and the application of new automation technology in the process of extraction traditional Chinese medicine in recent years were summarized, in order to provide a reference for Chinese medicine green and intelligent production.

Application of next generation sequencing for the analysis of patients with spontaneous abortion / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the value of next generation sequencing (NGS) for the analysis of spontaneous abortion samples.</p><p><b>METHODS</b>The NGS analysis was carried out on 85 chorionic villi samples (taken between 42 days to 12 weeks of gestation) for which conventional cell culture has failed or chromosomal karyotyping has yielded normal or uncertain result.</p><p><b>RESULTS</b>Among 68 samples with a normal karyotype, the NGS analysis has identified 2 copy number variations (CNVs) and 2 chimeras. For 16 cases with failed cell culture, the NGS has identified 4 chromosomal abnormalities including 1 copy number variation and 3 numerical chromosomal aberrations. For 1 remaining case with uncertain karyotyping result, the NGS analysis has verified it as 46,XX,del(4) (p15.1p16.3).seq[GRCh37/hg19] (57 549 - 32 371 364)×1.</p><p><b>CONCLUSION</b>The NGS analysis is capable of identifying novel CNVs in samples for which conventional cell culture may fail or karyotyping analysis may yield a normal result.</p>

A study on the risk factors for stroke among dialysis patients / 实用医学杂志

Objective To assess the mortality and risk factors for stroke among dialysis patients with different dialysis modality. Methods 590 patients who underwent hemodialysis (HD) or peritoneal dialysis (PD) from January 2008 to December 2012 were recruited in our study, and categorized according to dialysis modality. The prognostic risks of stroke were hazard ratio of risk was calculated by Cox regression analysis in HD and PD patients respectively. by the Kaplan?Meier curves or the Cox proportional hazards model. Results A total of 590 patients is under a median follow?up of 32.5 months. The stroke incidence rate of 49.2/1, 000 patient?years in total patients, and 74.1/1, 000 patient?years in HD patients, which was significantly higher compared with that of 31.8/1,000 patient?years in PD patients. On multivariate analysis, independent predictors of stroke occurrence were age(HR=1.05;95%CI:1.02~1.09;P=0.003)、diabete(HR=1.98;95%CI:1.31~3.46;P=0.001)、CVD(HR=2.06;95%CI:1.62-3.05;P < 0.001)、Total triglycerides(HR = 1.20; 95% CI:1.08-1.58; P = 0.034) and hemodialysis (HR = 2.03; 95% CI:1.46-3.89; P = 0.005). Conclusions Age, diabete, CVD, total triglycerides and hemodialysis are independently associated with increased stroke risks in dialysis patients, which suggest that these patients should pay attention to weight control and glucose control.

Clinical and genetic analysis for a patient with 45, X/46, X, Yqh- and mixed gonadal dysgenesis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the clinical and genetic characteristics of a patient with mixed gonadal dysgenesis.</p><p><b>METHODS</b>Clinical data was collected. The patient was subjected for serum hormone testing and G-banding chromosomal analysis. Sex-determining region of Y-chromosome (SRY) gene and azoospermia factor (AZF) a, b, c regions were analyzed with multiple polymerase chain reaction (PCR) and whole gene sequencing.</p><p><b>RESULTS</b>All serum hormone testing were normal. The karyotype of the patient was 45,X/46,X,Yqh-. PCR has proven the presence of SRY, ZFY and AZFa, and deletion of AZFb and AZFc regions. No mutation was detected in the sequence of the SRY gene. Abdominal computerized tomography has detected a huge mass in the pelvic cavity, which was positive for PLAP and CD117 on immunohistochemistry stain.</p><p><b>CONCLUSION</b>Based on clinical data and result of genetic testing, the patient was diagnosed with mixed gonadal dysgenesis. Pathological and immunohistochemistry analysis of the transformed gland has confirmed the diagnosis of seminoma. For patient with a karyotype of 45,X/46,X,Yqh-, the risk of seminoma may be related with the presence of SRY gene.</p>

Improved identification for trisomy 9p and partial trisomy 6q presented in a patient by array-based comparative genomic hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the genetic cause for a child with growth retardation and mental retardation and discuss the application of array-based comparative genomic hybridization (aCGH) in its molecular genetic diagnosis.</p><p><b>METHODS</b>Conventional karyotyping of peripheral blood for the family was carried out. aCGH was performed to further ascertain the size and origin of the additional chromosome fragments.</p><p><b>RESULTS</b>In the trio family here, the karyotype of the father was normal, the karyotype of the mother was 46,XX, t(6;9)(q26;q21)and the proband child's was 47,XX,+der(9)?t(6;9)(q26;q21). aCGH showed that the extra chromosomal fragments originated from chromosome 9p24.3-q21.13 and the size was 78.26 Mb, and the repeat region included the 9p trisomy's clinical area. At the same time, it was confirmed that 6q26-q27 was trisomic and the fragment that related to development delay was 6.6 Mb. We determined that the proband's karyotype was 47,XX,+der(9)t(6;9)(q26;q21.13)mat finally.</p><p><b>CONCLUSION</b>The patient's abnormal chromosome has originated from her mother with balance translocation. The duplications of 9p24.3-q21.13 and 6q26-q27 may lead to growth retardation and mental retardation. Accompanied with the cytogenetic methods, aCGH can accurately identify the origin and size of the abnormal chromosomes, contributing to the genetic analysis.</p>

Systematic genetic analysis for a case with duplication 9p / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify the genetic cause for a case with growth retardation and mental retardation.</p><p><b>METHODS</b>After conventional peripheral blood karyotyping with G-banding, the abnormal chromosome was identified as suspicious 9p duplication by multiplex ligation dependent probe amplification (MLPA) .</p><p><b>RESULTS</b>The proband's karyotype was suspicious 46,XY,der(9)t(9;14)(q13;q11.2), then the abnormal chromosome 9 was identified as 9p duplication with MLPA. The 9p duplication occurs because of a balanced chromosomal rearrangement between two chromosomes of 9 and 14 in the proband's father.</p><p><b>CONCLUSION</b>9p11.2-p24.3 duplication is the cause of abnormal phenotypes in the child patient. Cytogenetic methods combined with MLPA can efficiently identify abnormal chromosomes and provide accurate results for clinical diagnosis and treatment.</p>

Genetic analysis of a family with super-male syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify the genetic cause for a family featuring language retardation using combined cytogenetic and molecular genetic methods.</p><p><b>METHODS</b>Following conventional G-banded karyotype analysis, the additional Y chromosome was identified by fluorescence in situ hybridization (FISH) and multiplex ligation dependent probe amplification (MLPA). Whole genome array comparative genomic hybridization (aCGH) was also carried out to detect minor structural chromosomal abnormalities.</p><p><b>RESULTS</b>The proband's karyotype was determined as 47,XY,+?, and the unknown aberrant chromosome was identified as Yqh+ with FISH, MLPA and aCGH. No other chromosomal abnormality was found in the pedigree.</p><p><b>CONCLUSION</b>Cytogenetic methods combined with FISH, MLPA, and aCGH can efficiently identify the origin of unknown chromosomes and provide accurate clues for clinical diagnosis and treatment.</p>

Oxidative stress effect on viability of umbilical cord-derived mesenchymal stem cells in storage solution of transplantation / 中国组织工程研究

BACKGROUND:The viability of human umbilical cord-derived mesenchymal stem cel s is often declined with the commonly used transplantation storage solution in clinics, which may influence the therapeutic effects of cel ular transplantation. However, reasons for this are stil unknown. OBJECTIVE:To investigate the role of oxidative stress in the reduction of human umbilical cord-derived mesenchymal stem cel s viability in the storage process during clinical transplantation and to observe the effects of radical scavenger on the results. METHODS:Human umbilical cord-derived mesenchymal stem cel s were harvested and cultured in normal saline for 0, 2, 4 and 6 hours at room temperature. Intracel ular reactive oxygen levels were detected at those time points. Antioxidant enzyme activities and levels of malondialdehyde were measured to determine the intracel ular oxidative stress levels after storage. Cel adhesion rate changes were retested after adding N-acetyl cysteine to the storage solution. RESULTS AND CONCLUSION:The reactive oxygen levels in human umbilical cord-derived mesenchymal stem cel s were increased significantly after normal saline storage and levels of malondialdehyde were increased in a time-dependent manner. Activities of superoxide dismutase, catalase and glutathione peroxidase were al reduced. Addition of N-acetyl cysteine into the storage medium decreased the reactive oxygen levels and improved the human umbilical cord-derived mesenchymal stem cel s viabilities. Experimental findings indicate that, increased reactive oxygen species in human umbilical cord-derived mesenchymal stem cel s is one of the reasons for reduced cel viability. Adding the radical scavenger N-acetyl cysteine can improve the storage effects of human umbilical cord-derived mesenchymal stem cel s.

Evaluation indexes for the viability of umbilical cord-derived mesenchymal stem cells before transplantation / 中国组织工程研究

BACKGROUND:Umbilical cord-derived mesenchymal stem cel s are gaining more attention in clinical treatments. Cel viability prior to transplantation has a direct impact on clinical prognosis. Despite trypan blue staining is a widely performed procedure to assess the viability of umbilical cord-derived mesenchymal stem cel s, it cannot reflect the functional capacity of those cel s accurately because of some subjective factors. OBJECTIVE:To explore sensitive and accurate assay for the functions of umbilical cord-derived mesenchymal stem cel s. METHODS:Human umbilical cord-derived mesenchymal stem cel s were isolated and cultured in vitro. Cultured umbilical cord-derived mesenchymal stem cel s were preserved in 0.9%saline for 0, 2, 4 and 6 hours at 4 ℃. Various methods (trypan blue staining, AnnexinV-PI, terminal deoxynucleotidyl transferase dutp nick end labeling, cel counting kit-8, live-dead assay, cel adherent assay) were used to determine the viability of post-storage umbilical cord-derived mesenchymal stem cel s, and the results were compared with colony-forming efficiency, a measure of cel function. RESULTS AND CONCLUSION:Human umbilical cord-derived mesenchymal stem cel s cultured in vitro showed a spindle shape and attached growth, the third-generation umbilical cord-derived mesenchymal stem cel s were positive for CD29, CD44, CD105, and negative for CD 34 and CD 45. Umbilical cord-derived mesenchymal stem cel s incubated in the adipogenic and osteogenic medium were both positive. Cel viability measured with trypan blue correlated moderately with colony-forming efficiency, while the percentage of viable cel s measured with other methods correlated better with colony-forming efficiency, among which adherent assay was the most obvious. It is proved that cel adherent assay-measured viability is the most accurate indicator.

Influence of post-discharge continuous nursing intervention on psychological quality and compliance with immunosuppressants treatment in patients with renal transplantation / 中国实用护理杂志

Objective To explore the effects of postdischarge follow-up continuous nursing intervention on the psychological quality and compliance of immunosuppressants treatment in patients with kidney transplantation.Methods 157 patients with kidney transplantation post-operation were recruited,81 cases in the study group and 76 cases in the control group randomly.The control group received routine hospital discharge care and immunosuppressants treatment.The study group received postdischarge followup nursing intervention and immunosuppressants treatment.After 6 months of postdischarge intervention,the psychological quality of patients in two groups was assessed by the emotion application measure scale separately.The Morisky Medication Adherence Scale 8-item version (MMAS 8-item version) was used for the assessment of the immunosuppressants treatment compliance of the two groups of patients.Results After 6-month intervention,the patients in the study group showed better psychological quality than the patients in the control groups.There were significant differences in the various items of assessments of the psychological quality between the two groups.The high compliance percentage of immunosuppressants treatment of the study group was 45.7％,whereas the high compliance percentage of the control group was 22.4％.The differences between the two groups were significant.Conclusions Post-discharge follow-up nursing intervention to patients with kidney tranplantantion is effective for improving the psychological quality and the immunosuppressants treatment compliance and may be helpful to improve the life quality of the patients.

Proteomic identification of CIB1 as a potential diagnostic factor in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC), among the most common malignancies worldwide, remains a major threat to public health, and there is an urgent need to identify novel biomarkers for diagnosis, prognosis and targets for anti-cancer treatment. In this study, two-dimensional polyacrylamide gel electrophoresis coupled with ESI-Q-TOF MS/MS analysis was used to identify differentially expressed proteins among the HCC tumour centre, tumour margin and nontumourous liver tissues. In total, 52 spots with significant alteration were positively identified byMS/MSanalysis. Altered expression of representative proteins, including CIB1, was validated by Western blotting. Immunostaining suggested an increase tendency of CIB1 expression from nontumourous liver tissue to tumour centre. Knockdown of CIB1 expression by RNA interference led to the significant suppression of the cell growth in hepatoma HepG2 cells. These data suggest that CIB1 may be used as a novel prognostic factor and possibly an attractive therapeutic target for HCC.

Repair of acute anterior cruciate ligament injury usingpolyethylene terephthalates LARS artificial ligament in 23 cases / 中国组织工程研究

BACKGROUND:LARS artificial ligament was designed by Laboureau from France using polyethylene terephthalates with the anatomic structure of human ligament and mechanical principle of weight.This ligament is not only accorded with physiological structure of normal anterior cruciate ligament,but also significantly elevates anti-torque,can resist repetitive twist,bend and excessive traction.OBJECTIVE:To summarize characteristics of clinical application of LARS artificial ligament in transplant reconstruction following acute anterior cruciate ligament of knee joint injury.METHODS:A total of 23 patients with acute anterior cruciate ligament injury,comprising 17 males and 6 females,aged 21-54 years,were selected.Time-from injury to surgery was 3 days to 3 weeks.Polyethylene terephthalatas LARS artificial ligament made in France was used to reconstruct damaged ligament.The transplant was evaluated before and after implantation according to Lysholm knee joint criteria.RESULTS AND CONCLUSION:Following 11.2 months (10-14 months) of follow-up,unstable symptoms of affected knees of all 23 cases disappeared.Anterior and posterior drawer tests were negative,with good joint function.Average extension and flexion degree was 0°-120°.In accordance with Lysholm knee joint score,significant difference was found from (40.34±4.00) points preoperatively to (90.21±4.00) points postoperatively (P<0.01).They could do common athletic activities at about 2 months following surgery.In some cases,magnetic resonance examination demonstrated that residue ligament gradually grew into artificial ligament,which gradually became thicker.Above-mentioned results have verified that under an arthroscope,reconstruction of anterior cruciate ligament using polyethylene terephthalates LARS artificial ligament showed simple operation and small wound,which may lead to immediate stability,early rehabilitation exercises.Simultaneously,interlacing of residue ligament and artificial ligament keeps the nerve conduction pathway of propdoceptive sense,which prevents joint function limitation greatly.Moreover,short-term outcomes are satisfactory.

Relationship between Talin and its signaling pathway and hepatocellular carcinoma / 国际外科学杂志

Alterations of cell adhesion of tumor cells is the initial step of invasion and metastasis of tumor cells.Integrin receptors are cell surface receptors with critical functions in cell adhesion and migration,which can mediate many signaling pathway occurred,including the integrin/FAK signaling pathway,directly or indirectly affecting tumor recurrence and metastasis.Integrin/FAK signaling pathway is thought to play an impartant role in integrin-mediated signaling transduction pathway leading to adhesion and metastasis of hepatocellular carcinoma.Talin is the first cytoskeletal protein that has been proposed to act as the final common step in integrin activation,which is a major component of the focal adhesion.This study reviews the construction and function of talin,its reaction with integrin/FAK signaling pathway,as well as its relationship with hepatocellular carcinoma and other tumors.

Purification and characterization of deoxyribonuclease from earthworm Eisenia foetida / 北京大学学报(医学版)

Objective:To purify a kind of deoxyribonuclease from earthworm Eisenia foetida (named earthworm DNase, EDNase) and study its characteristics. Methods: Acetone precipitation, ion-exchange chromatography, high performance liquid chromatography, SDS-PAGE, CapiUary electrophoresis isoelectric focusing and MALDI-TOP MS were used for the study. Results: This purified protocol improved 137-fold purification and 45.6 % recovery of enzyme activity. The molecular mass of EDNase was estimated to be 63 000. Mg2+ , Mn2+ and Ca2+ were strong inhibitors of EDNase, while Na+ slightly increasd the enzyme activity. The enzyme was completely stable in the pH range from 4. 4 to 5.2 and had a pH optimum of 4.8. The optimum temperature was 37℃ and the enzyme was stable up to 40 ℃. The pI of the enzyme was 6. 20. Km and Vmax for the enzyme were 1.52 g/L and 4. 89 mg/(mL ·min), respectively, with calf thymus DNA as substrate. The enzyme was able to degrade chromosemal DNA, linear λbacteriophage DNA as well as supereoiled plasmid DNA, but didn' t display any RNase activity. Conclusion: This kind of deoxyribonuclease possesses unique characteristics, which is different from the deoxyribonucleases which we have known before.

A KINETICS STUDY OF DIFFERENTSPECIES OFBRUCELLA ON THE SUPPRESSIVE EFFECT OF PFC IN C57/BL MICE / 中国免疫学杂志

Data showed that there were Suppressive effects of B.melitensis 16,B.abortus 544 and B.suis 12 on PFC in different degree in C_(57)/BL.Thisimmunosuppressive effect was increased with the time of infection and ma-intained untill 6 months after infection while the Brucella already wasfully eradicated.According to the data,authors suggested that there was alead from the view of immunosuppression in the study of pathogenesis ofBrucellosis except the bacterium and allergy.