Objective To analyze the karyotypes of 11 cases of Turner syndrome with marker chromosome,and study the phenotypic effects resulting from the abnormal karyotype.Methods Eleven Turner syndrome patients had a mosaic karyotype and carried a marker chromosome,and 6 marker chromosomes were ring chromosomes.Their karyotypes were showed as mos.45,X/46,X,+mar or mos. 45,X/46,X,+r.Fluorescence in situ hybridization(FISH)technique with X/Y centromere probes was performed to determine the origin of the marker chromosome.Reverse chromosome painting technique was used to identify the breakpoints of two largest markers.Phenotype effects with different chromosome breakpoints were compared.Results All the 11 marker chromosomes were ring X chromosomes.The breakpoints of the r(X)were involved in Xp22,Xq22,Xq24 and Xq26,etc.Conclusions The marker chromosomes in Turner syndrome mainly originate from X chromosome and form ring chromosome X.Each r (X)in our patients was mosaic,indicating it was originated from mitosis error during early embryo development.To analyze the origin of the marker chromosome and the breakpoint of r(X)will provide guidance for the therapy and prognosis of the Turner syndrome patient.

To investigate the effect and possible mechanism of echinacoside-containing serum on the osteogenic differentiation in rat bone marrow mesenchymal stem cells. Rat bone marrow mesenchymal stem cells were cultivated by the whole bone marrow adherence method. The 3rd generation of cells were divided into 3 groups: the blank control group, the classic osteogenic-induced group and the 10% echinacoside-containing serum group. The expression of alkaline phosphatase and osteocalcin were detected by ELISA. The ex- pression of ZHX, protein was detected by Western blot technique. RT-PCR technique was used to detect the expression of ZHX₃mRNA. According to the result, the expressions of the alkaline phosphatase and osteocalcin in the classic osteogenic-induced group and the 10% echinacoside-containing serum group were significantly higher than that of the blank control group (P <0. 01). And expressions of the alkaline phosphatase activity and osteocalcin in the 10% echinacoside-containing serum group were significantly higher than that in the classic osteogenic-induced group (P < 0.01). Meanwhile, the classic osteogenic-induced group and the 10% echinacoside-containing serum group showed obviously higher ZHX₃ protain and mRNA expression than that of the black control group, with significant differences (P < 0.01); the 10% echinacoside-containing serum group showed obviously higher ZHX₃ protain and mRNA expression than that of the classic osteogenic-induced group, with a significant difference (P < 0.01). In conclusion, 10% echinacoside-containing serum can promote the differentiation of the bone marrow mesenchymal stem cells cultured in vitro. Its mechanism may be correlated with the increase in the ZHX₃expression.
Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Glycosides ; blood ; pharmacology ; Homeodomain Proteins ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Osteogenesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Serum ; chemistry ; Transcription Factors ; genetics ; metabolism

Recent researches indicate that ubiquitin-protea some pathway plays an important role in apoptosis regulation. Proteasome inhibitors induce apoptosis in many kinds of neoplastic cells, thus provide a great opportunity for exploring synergy of proteasome inhibitors and other apoptosis-inducing agents. In this study, the effect of the proteasome inhibitor Z-LLL-CHO combined with etoposide (VP16) on leukemic cell lines M-07e and TF-1 was investigated by MTT assay, trypan blue exclusion, flow cytometry and Western blot. The results showed that the combination of Z-LLL-CHO and VP16 was much more effective than either agents alone in promoting cytotoxicity in both cell lines evaluated. Accumulation of cells in S + G2/M phase of the cell cycle was observed in the cells treated with VP16 and Z-LLL-CHO alone, while apparent increase of sub-G0/G1 fraction was detected in cells treated with combination of the agents. The cleavage of Bcl-2 into a shortened 22 kD fragment was detected in M-07e cells exposed to either agents alone, and the fraction of 22 kD fragment was increased in the cells treated with combination of the agents. In conclusion, the combination of Z-LLL-CHO and VP16 enhanced their individual cytotoxic effect by inducing apoptosis, in which increase of S + G2/M fraction in cell cycle as well as the enhanced cleavage of Bcl-2 are the possible mechanism of the additive effect on leukemic cells by Z-LLL-CHO and VP16.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cysteine Endopeptidases ; Etoposide ; pharmacology ; Humans ; Leukemia ; pathology ; Multienzyme Complexes ; antagonists & inhibitors ; Oligopeptides ; pharmacology ; Proteasome Endopeptidase Complex ; Proto-Oncogene Proteins c-bcl-2 ; analysis

OBJECTIVETo determine the karyotype of a patient with Prader-Willi-like syndrome features.

METHODSChromosomal high resolution banding was carried out to analyze the karyotype of the patient, and methylation-specific PCR was used to analyze the imprinting region of chromosome 15. Subtelomeric region was screened by multiplex ligation-dependent probe amplification (MLPA), and fluorescent in situ hybridization (FISH) and real-time quantitative PCR were further performed to identify the deleted region.

RESULTSNo abnormality was discovered by high resolution karyotype analysis and methylation-specific PCR studies. MLPA analysis showed that the patient had a deletion of 1p subtelomeric area, which was confirmed by FISH analysis. The deleted region was shown within a 4.2 Mb in the distal 1p by 3 BAC FISH probes of 1p36 combined with real-time PCR technique. Family pedigree investigation showed the chromosome abnormality was de novo. Therefore, partial monosomy 1p36 was likely responsible for the mental retardation of the patient.

CONCLUSIONMolecular cytogenetic techniques should be performed to those patients with Prader-Willi-like syndrome features, to determine their karyotypes.

Child ; Chromosome Deletion ; Chromosomes, Human, Pair 1 ; genetics ; Female ; Humans ; Karyotyping ; Prader-Willi Syndrome ; genetics

OBJECTIVETo establish a single-cell whole genome amplification (WGA) technique, in combination with comparative genomic hybridization (CGH), for analyzing chromosomal copy number changes, and to explore its clinical application in preimplantation genetic diagnosis (PGD).

METHODSTwelve single-cell samples with known karyotypes, including 5 chorionic villus samples, 4 human embryonic stem cell (hESC) samples and 3 peripheral lymphocyte samples, and 4 single blastomere samples carrying chromosomal abnormalities detected by PGD, were collected for whole genome amplification by combining primer extension preamplification (PEP) with degenerate oligonucleotide primed-PCR (DOP-PCR) amplification. The amplified products labeled by red fluorescence were mixed with control DNA labeled by green fluorescence, and then the mixture was analyzed by CGH. As a comparison, 10 single cell samples were amplified by DOP-PCR only and then CGH analysis was performed.

RESULTSThe amplification using PEP-DOP-PCR was more stable than traditional DOP-PCR. The products of PEP-DOP-PCR range from 100 bp to 1000 bp, with the mean size being about 400 bp. The CGH results were consistent with analyses by other methods. However, only 6 out of 10 single cell samples were successfully amplified by DOP-PCR, and CGH analysis showed a high background and 2 samples showed inconsistent results from other methods.

CONCLUSIONPEP-DOP-PCR can effectively amplify the whole genome DNA of single cell. Combined with CGH, this WGA method can successfully detect single-cell chromosomal copy number changes, while DOP-PCR was easy to fail to amplify and amplify inhomogeneously, and CGH analysis using this PCR product usually showed high background. These results suggest that PEP-DOP-CGH is a promising method for preimplantation genetic diagnosis.

Comparative Genomic Hybridization ; methods ; DNA Primers ; Genetic Testing ; methods ; Humans ; Karyotyping ; methods ; Nucleic Acid Amplification Techniques ; methods ; Nucleic Acid Hybridization ; methods ; Oligonucleotides ; chemistry ; Preimplantation Diagnosis ; methods

Objective To investigate the distribution of insulin resistance (HOMA-IR) index and its influencing factorsamong middle and old aged people with normal glucose and to provide the basis for early screening and prevention of type 2diabetes. Methods A total of 229 residents were selected with health records showed normal blood glucose (fasting bloodglucose < 7.0mmol/L, postprandial 2h blood glucose<11.1 mmol/L) and more than 40 years old from July, 2012 to June,2015. Height, weight, waist and hip circumference, and the fasting plasma glucose (FPG), insulin (FINS), lowdensity lipoprotein (LDL), uric acid, tumor necrosis factor (TNF) and interleukin -6 (IL-6) were recorded to analyzethe distribution of HOMA-IR and its influencing factors. Results Totally 229 people were included, of which 113 were male(49.34%), 116 female(50.66%) . The average age was(63.58 + 8.85) years old. The average HOMA-IR index was 0.94(1.08) and there were 21 people that HOMA-IR exceed the standard (HOMA-IR≥2.68), accounting for 9.17%.TheHOMA-IR index of different gender, age, waist circumference, hip circumference, uric acid in the elderly had significantdifference (P < 0.05) .Multiple linear regression analysis showed that HOMA-IR index was positively correlated withfemale, waist circumference and IL-6 and was negatively correlated with age. Conclusion The possibility of IR was higherin women with relatively low age, female, central obesity and high IL-6 levels among the middle and old aged people withnormal blood glucose.

OBJECTIVETo identify the role of 5alpha-reductase in the spermatogenesis of male rats by studying the effect of two 5alpha-reductase inhibitors, Epristeride and Finasteride, on the spermatogenesis in male Sprague-Dawley (SD) rats.

METHODSChanges in the weight of the testis, serum testosterone and dihydrotestosterone levels, epididymal sperm count, and reproductive function were observed and analyzed after the two 5alpha-reductase inhibitors were administered to male SD rats orally.

RESULTSThe experiment showed that in comparison with control animals, both the two 5alpha-reductase inhibitors: 1. suppressed the development of the prostate and reduced the weight of the testis in the experimental groups (P < 0.05); 2. decreased the serum level of dihydrotestosterone and enhanced testosterone; 3. inhibited epididymal sperm count and productive function.

CONCLUSIONHigh dosages of the 5alpha-reductase inhibitor, Epristeride, can suppress the development of the prostate and reduce the weight of the testis, decrease dihydrotestosterone, and inhibit spermatogenesis and productive function in male rats.

5-alpha Reductase Inhibitors ; Androstadienes ; pharmacology ; Animals ; Dose-Response Relationship, Drug ; Finasteride ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Spermatogenesis ; drug effects

OBJECTIVETo investigate the clinical feasibility of cell-free fetal DNA (cffDNA)-based noninvasive prenatal diagnosis of β-thalassemia.

METHODSNine samples of amniotic fluid were obtained to detect the 8 common and 9 relatively rare mutation sites of β-thalassaemia in Guangdong Province. The maternal blood samples were also collected for extracting and purification of the cffDNA, and a duplex PCR was performed using 3 pairs of primers and the fetal β-globin genotype was analyzed by reverse dot-blot hybridization.

RESULTSAmong the 9 cases, 5 showed fetal genotypes of β-thalassemia inherited from the father by examination of the amniotic fluid, and 2 fetuses were identified to have β-thalassemia genes inherited from the father determined based on the cffDNA in the maternal blood.

CONCLUSIONSThe cffDNA-based noninvasive prenatal diagnosis is feasible for β-thalassemia, but the contamination of the maternal background DNA results in a low detection rate.

Adult ; Cell-Free System ; DNA ; blood ; Female ; Fetal Diseases ; diagnosis ; genetics ; Fetus ; Genetic Testing ; Humans ; Pregnancy ; blood ; Prenatal Diagnosis ; methods ; Young Adult ; beta-Thalassemia ; diagnosis ; genetics

·AMI:To evaluate the clinical efficacy of the segmented bifocal intraocular lens (MIOL) SBL-3. ·METHODS:Totally 26 cases (26 eyes) of age-related cataract who received phacoemulsification and implantation of MIOL SBL-3 were enrolled from February 2016 to June 2017 in our hospital as the SBL-3 group. And 28 cases (28 eyes) of age-related cataract who received phacoemulsification and implantation of single focus intraocular lens intraocular lens TecnisZA9003 were enrolled as control group. At postoperative 3mo, the uncorrected visual acuity, corrected visual acuity, contrast sensitivity and patient satisfaction were compared between two groups. ·RESULTS: At preoperative, there was no statistical difference in uncorrected distance, intermediate and near visual acuity between two groups (P > 0. 05). At postoperatively 3mo, the uncorrected distance, intermediate and near visual acuity in patients of both groups were significantly improved (P < 0. 05). At postoperative 3mo,the uncorrected intermediate and near visual acuity, distance-corrected intermediate and near visual acuity, contrast sensitivity at different spatial frequencies (3, 6, 12, 18c/d) and patient satisfaction in SBL- 3 group were significantly better than those of control group (P<0.05). ·CONCLUSION:The MIOL SBL-3 not only could provide preferable distance visual acuity, but also could provide better intermediate visual acuity, near visual acuity and contrast sensitivity,and greatly increase the visual quality and satisfaction at postoperative in cataract patients.

·AIM: To compare the visual quality after different multifocal lens (MIOL) implantation in cataract patients. ·METHODS: Totally 78 cases (78 eyes) of cataract patients who receiving phacoemulsification and implantation of different MIOL were enrolled from February 2016 to June 2017 in our hospital. According to the different type of implanted MIOL, the 78 cases (78 eyes) were divided Group A,B and C,each group was 26 cases (26 eyes). Patients in the Group A were implanted regional refraction MIOL SBL-3; patients in the Group B implanted step progressive diffraction MIOL SN6AD1;patients in the Group C implanted all - optical plane diffraction MIOL Tecnis ZMB00. The visual acuity, contrast sensitivity,defocus testing,contrast visual acuity and satisfaction of visual quality were compared in 3 groups at postoperative 3mo. · RESULTS: There was no statistically significant difference in postoperative adverse reactions between the three groups (P>0.05). There was statistical difference in uncorrected intermediate visual acuity, uncorrected near visual acuity, distance - corrected intermediate visual acuity and distance- corrected near visual acuity in 3 groups (P < 0. 05), and those in the Group C were significantly worse than those of Group A and B (P<0.05). There was statistical difference in contrast sensitivity at four spatial frequencies (3,6,12,18c/d) in light and light glare and three spatial frequencies (3, 6, 12c/d) in dark and dark glare in 3 groups (P<0.05), and those in the Group A were significantly better than those of Group B and C (P<0.05). According to the defocus testing of 3 groups,the visual acuity at -1.5D,-2.0D and-2. 5D in Group A and B was significantly higher than Group C (P<0. 05). There was statistical difference in contrast visual acuity at 20% and 9% of contrast ratio in 3 groups (P<0.05),and those in Group A were significantly better than Group B and C(P<0.05). There was statistical difference in visual quality satisfaction in 3 groups (P<0.05), and that in the Group A was significantly better than that of Group B and C (P<0.05). ·CONCLUSION:The region refraction MIOL SBL-3 not only could provide better distant, intermediate and near visual acuity, but also could provide better contrast sensitivity and contrast visual acuity, thereby greatly increase visual quality satisfaction.