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1.
Reproduction ; 168(2)2024 Aug 01.
Article in English | MEDLINE | ID: mdl-38819377

ABSTRACT

In brief: Ovarian aging results in reactive oxygen species accumulation and mitochondrial deterioration. During the aging process, GRSF1 deficiency attenuates mitochondrial function in aging granulosa cells. Abstract: Ovarian aging critically influences reproductive potential, with a marked decrease in oocyte quality and quantity and an increase in oxidative stress and mitochondrial dysfunction. This study elucidates the role of guanine-rich RNA sequence binding factor 1 (GRSF1) in the aging of ovarian granulosa cells (GCs). We observed a significant reduction in GRSF1 within GCs correlating with patient age, utilizing clinical samples from IVF patients. Using an siRNA-mediated knockdown technique, we established that diminished GRSF1 expression exacerbates mitochondrial dysfunction, elevates reactive oxygen species, and impairs ATP production. Furthermore, RNA immunoprecipitation revealed GRSF1's interaction with superoxide dismutase 2 (SOD2) mRNA, a key antioxidant enzyme, suggesting a mechanism whereby GRSF1 modulates oxidative stress. Downregulation of SOD2 reversed the protective effects of GRSF1 overexpression on mitochondrial function. These insights into the role of GRSF1 in ovarian aging may guide the development of interventions to improve fertility outcomes in advanced age.


Subject(s)
Aging , Cellular Senescence , Granulosa Cells , Mitochondria , Oxidative Stress , Reactive Oxygen Species , Female , Granulosa Cells/metabolism , Humans , Mitochondria/metabolism , Reactive Oxygen Species/metabolism , Aging/metabolism , Adult , Superoxide Dismutase/metabolism , Superoxide Dismutase/genetics , Cells, Cultured , Poly(A)-Binding Proteins
2.
Heliyon ; 9(9): e19395, 2023 Sep.
Article in English | MEDLINE | ID: mdl-37809636

ABSTRACT

Objective: Zishen Yutai Pill (ZYP) is a frequently used traditional Chinese medicine (TCM) preparation in women's health. However, the effects of ZYP on endometrial epithelial response have not been fully explored. Herein, uterine natural killer cell (uNK) secretion medium was used to mimic the uterine microenvironment. Thereafter, an endometrial epithelial cell line (Ishikawa cells) was treated with ZYP-containing serum to elucidate the effects of ZYP on endometrial receptivity.Methods: uNK cells were isolated from decidual tissues of pregnant women undergoing pregnancy termination surgery, and thereafter, uNK secretion medium was collected. ZYP-containing serum was collected from rats after intragastrical administration of ZYP. Ishikawa cells were divided into three groups, one treated with blank control (control group), one treated with uNK secretion medium (uNK group), and one treated with both uNK secretion medium and ZYP-containing serum (ZYP + uNK group). Total RNAs were extracted. Gene expression profiles of Ishikawa in different groups were determined through microarray analysis. mRNA expressions of selected genes were determined through quantitative real-time polymerase chain reaction (qRT-PCR). Expression of intercellular cell adhesion molecule-1 (ICAM-1) was determined using Western blotting (WB). Results: Compared with the uNK group, the gene expressions of ZYP group with a total of 1117 genes were significantly altered, among which 510 genes were upregulated and 607 genes were downregulated. Compared with uNK group, expressions of CSF1, CSF2, SPP1, and ICAM1 were upregulated (P < 0.05). Up-regulation of ICAM-1 expression after treatment of ZYP was further confirmed by WB analysis. Conclusion: In brief, in the presence of uNK cell medium, ZYP could improve the expressions of ICAM1, CSF1, CSF2, TNF, SPP1, etc. However, further exploration should be carried out in in vivo experiments for the validation of the mechanisms of ZYP on endometrial epithelial response.

3.
Insect Sci ; 28(6): 1690-1707, 2021 Dec.
Article in English | MEDLINE | ID: mdl-33118290

ABSTRACT

Validamycin has been widely used as a specific competitive inhibitor of trehalase. In our previous research, validamycin significantly inhibited trehalase activity and chitin synthesis in Diaphorina citri, resulting in abnormal phenotypes. However, the mechanism of validamycin's action on D. citri remains unclear. Here, using a comparative transcriptome analysis, 464 differentially expressed genes (DEGs) in D. citri were identified after validamycin treatment. A Gene Ontology enrichment analysis revealed that these DEGs were mainly involved in "small molecule process", "structural molecule activity" and "transition metal ion binding". DEGs involved in chitin metabolism, cuticle synthesis and insecticide detoxification were validated by reverse transcription quantitative polymerase chain reaction. The RNA interference of D. citri chitinase-like protein ENO3 and D. citri cuticle protein 7 genes significantly affected D. citri molting. Moreover, the recombinant chitinase-like protein ENO3 exhibited a chitin-binding property, and an antimicrobial activity against Bacillus subtilis. This study provides a first insight into the molecular changes in D. citri after exposure to validamycin and identifies two effective RNA interference targets for D. citri control.


Subject(s)
Chitinases , Hemiptera , Inositol/analogs & derivatives , RNA Interference , Transcriptome , Animals , Chitin/biosynthesis , Chitinases/antagonists & inhibitors , Chitinases/genetics , Hemiptera/drug effects , Hemiptera/genetics , Hemiptera/metabolism , Inositol/pharmacology
4.
J Colloid Interface Sci ; 579: 562-572, 2020 Nov 01.
Article in English | MEDLINE | ID: mdl-32623122

ABSTRACT

To improve the permeability and antifouling properties of polyvinyl chloride (PVC) ultrafiltration (UF) membrane, amphiphilic sulfonated polysulfone (SPSF) was introduced into PVC matrix. Three types of PVC/SPSF blend membranes containing different SPSF with the sulfonation degree (SD) of 20%, 30%, and 50% were fabricated by non-solvent induced phase separation (NIPS) process. The excellent compatibility between PVC and SPSF was confirmed by differential scanning calorimetry (DSC). Surface chemical compositions, morphology, roughness, charge, hydrophilicity, permeability and antifouling properties of the pristine PVC membrane and the PVC/SPSF blend membranes were systematically compared and characterized. Due to the improved hydrophilicity and endowed negative charge, the blend membrane showed high water permeability (i.e. 880 L m-2h-1 bar-1), high bovine serum albumin (BSA) rejection (i.e. 95.7%), and high flux recovery ratio (i.e. 96%), which outperformed ever reported and commercialized PVC membranes. Furthermore, the permeability and rejection properties of PVC/SPSF UF membranes were maintained after soaking in acidic and alkaline solutions for 30 days, indicating their outstanding acid and alkali tolerance. Therefore, SPSF was expected as a potential versatile modifier for fabricating high performance UF membranes.

5.
Insects ; 11(4)2020 Apr 17.
Article in English | MEDLINE | ID: mdl-32316461

ABSTRACT

Chitin is one the main components of the insect cuticle, and chitin synthase (CHS) is an important enzyme required for chitin formation. CHS has been characterized in various insect species, but the structure and biochemical properties in Spodoptera litura have not been determined. In this study, we identified two CHS genes, SlCHS1 and SlCHS2, which encode proteins with 1565 and 1520 amino acid residues, respectively. Transcriptional analysis suggested that SlCHS1 has a high expression level in the integument whereas SlCHS2 showed the highest expression level in the midgut. During S. litura growth and development, SlCHS1 and SlCHS2 were both predominantly expressed in the fourth-instar larval stage. In addition, the expression of SlCHS1 and SlCHS2 could be induced by 20-hydroxyecdysone (20E). Silencing of SlCHS1 by RNA interference significantly inhibited the pupation and molting of S. litura larvae (RNAi), while knockdown of SlCHS2 had no significant effects on the S. litura phenotype. These results may provide a new molecular target for control of S. litura.

6.
Appl Opt ; 59(6): 1611-1617, 2020 Feb 20.
Article in English | MEDLINE | ID: mdl-32225659

ABSTRACT

A theoretical model on accumulation of temperature and stress in a three-junction GaAs solar cell is proposed to analyze its damage characteristics while irradiated by a multipulse laser. The distribution and accumulation effect of temperature and stress with different pulse widths are calculated. Specifically, the influences of pulse energy and duty ratio on the accumulation effect are discussed. Results show that the accumulation is weakened as pulse energy and duty ratio decrease and differ with the different pulse widths. The accumulation rate of stress is more rapid than that of temperature under nanosecond laser irradiation. Furthermore, tensile stress damage is the main damage form under nanosecond laser irradiation, and melting damage will change to main damage from a millisecond laser.

7.
RSC Adv ; 10(48): 28720-28724, 2020 Aug 03.
Article in English | MEDLINE | ID: mdl-35520074

ABSTRACT

A highly diastereoselective dearomative [3 + 2] 1,3-dipolar cycloaddition reaction of nitrobenzothiophenes with an in situ-generated nonstabilized azomethine ylides has been developed. The transformation provides a series of functionalized fused tricyclic benzo[4,5]thieno[2,3-c]pyrroles in good yields (up to 92%) under mild reaction conditions. In addition, a gram-scale experiment and the synthetic transformation of the cycloadduct further highlighted the synthetic utility. The relative configurations of the typical products were clearly confirmed by X-ray crystallography.

8.
Int J Mol Sci ; 21(1)2019 Dec 20.
Article in English | MEDLINE | ID: mdl-31861829

ABSTRACT

Chitin deacetylase (CDA) is a chitin degradation enzyme that strictly catalyzes the deacetylation of chitin to form chitosan, which plays an important role in regulating growth and development, as well as the immune response. In this study, a chitin deacetylase 3 gene (CDA3) was identified with a complete open reading frame (ORF) of 1362 bp from the genome database of Diaphorina citri, encoding a protein of 453 amino acids. Spatiotemporal expression analysis suggested that D. citri CDA3 (DcCDA3) had the highest expression level in the integument and third-instar nymph stage. Furthermore, DcCDA3 expression level can be induced by 20-hydroxyecdysone (20E). Injection of Escherichia coli and Staphylococcus aureus induced the upregulation of DcCDA3 in the midgut, while DcCDA3 was downregulated in the fat body. After silencing DcCDA3 by RNA interference, there was no influence on the D. citri phenotype. In addition, bactericidal tests showed that recombinant DcCDA3 inhibited gram-positive bacteria, including S. aureus and Bacillus subtilis (B. subtilis). In conclusion, our results suggest that DcCDA3 might play an important role in the immune response of D. citri.


Subject(s)
Amidohydrolases/immunology , Hemiptera/immunology , Insect Proteins/immunology , Amidohydrolases/chemistry , Amidohydrolases/genetics , Amino Acid Sequence , Animals , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/immunology , Hemiptera/chemistry , Hemiptera/genetics , Immunity , Insect Proteins/chemistry , Insect Proteins/genetics , Transcriptome
9.
Int J Mol Sci ; 20(15)2019 Jul 31.
Article in English | MEDLINE | ID: mdl-31370145

ABSTRACT

Chitin synthase is a critical enzyme that catalyzes N-acetylglucosamine to form chitin, which plays an important role in the growth and development of insects. In this study, we identified a chitin synthase gene (CHS) with a complete open reading frame (ORF) of 3180 bp from the genome database of Diaphorina citri, encoding a protein of 1059 amino acid residues with the appropriate signature motifs (EDR and QRRRW). Reverse transcription-quantitative PCR (RT-qPCR) analysis suggested that D. citri CHS (DcCHS) was expressed throughout all developmental stages and all tissues. DcCHS had the highest expression level in the integument and fifth-instar nymph stage. Furthermore, the effects of diflubenzuron (DFB) on D. citri mortality and DcCHS expression level were investigated using fifth-instar nymph through leaf dip bioassay, and the results revealed that the nymph exposed to DFB had the highest mortality compared with control group (Triton-100). Silencing of DcCHS by RNA interference resulted in malformed phenotypes and increased mortality with decreased molting rate. In addition, transmission electron microscopy (TEM) and fluorescence in situ hybridization (FISH) also revealed corresponding ultrastructural defects. Our results suggest that DcCHS might play an important role in the development of D. citri and can be used as a potential target for psyllid control.


Subject(s)
Chitin Synthase/genetics , Genome, Insect , Hemiptera/genetics , Insect Proteins/genetics , Nymph/genetics , RNA Interference , Amino Acid Sequence , Animals , Chitin Synthase/antagonists & inhibitors , Chitin Synthase/metabolism , Citrus/parasitology , Diflubenzuron/pharmacology , Fruit/parasitology , Gene Expression Regulation, Developmental , Hemiptera/drug effects , Hemiptera/enzymology , Hemiptera/growth & development , Insect Control , Insect Proteins/antagonists & inhibitors , Insect Proteins/metabolism , Molting/drug effects , Molting/genetics , Nymph/drug effects , Nymph/growth & development , Nymph/metabolism , Open Reading Frames , Phylogeny , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Sequence Alignment , Sequence Homology, Amino Acid
10.
Arch Insect Biochem Physiol ; 102(1): e21592, 2019 Sep.
Article in English | MEDLINE | ID: mdl-31276235

ABSTRACT

Ferritin, which is ubiquitous among all living organisms, plays a crucial role in maintaining iron homeostasis, immune response, and detoxification. In the present research, we identified an iron-binding protein, ferritin heavy chain subunit, from Papilio xuthus and named PxFerHCH. The complete complementary DNA of PxFerHCH was 1,252 bp encoding a sequence of 211 amino acids, which includes an iron-responsive element. Phylogenetic analysis showed that PxFerHCH is clustered with Manduca sexta and Galleria mellonella ferritin heavy chain subunits. Expression levels of PxFerHCH in various tissues were analyzed by reverse transcription quantitative polymerase chain reaction, and the results exhibited that PxFerHCH was expressed in all tissues with the highest expression in the fat body. The relative expression level of PxFerHCH in response to bacterial (Escherichia coli and Staphylococcus aureus) challenges sharply increased by about 12 hr postinfection (hpi) and then decreased at 24 hpi. In addition, the iron-binding capacity and antioxidation activity of recombinant PxFerHCH protein were also investigated. These results reveal that PxFerHCH might play an important role in defense against bacterial infection.


Subject(s)
Apoferritins/metabolism , Butterflies/metabolism , Iron/metabolism , Amino Acid Sequence , Animals , Apoferritins/genetics , Apoferritins/isolation & purification , Base Sequence , Butterflies/genetics , Butterflies/immunology , Escherichia coli , Insect Proteins/genetics , Insect Proteins/metabolism , Staphylococcus aureus
11.
J Insect Physiol ; 114: 125-135, 2019 04.
Article in English | MEDLINE | ID: mdl-30817914

ABSTRACT

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is the transmitting vector of Candidatus Liberibacter asiaticus (CLas), which causes citrus disease Huanglongbing (HLB). In recent years, control of HLB has been achieved by reducing the vector population. In the present study, we identified an isoform of D. citri tropomyosin (herein designated as DcTm1-X1). DcTm1-X1 was down-regulated in CLas-infected ACPs compared with uninfected ACPs. Bioinformatics analysis revealed that the full-length DcTm1-X1 is 2955 bp and encodes a protein of 284 amino acids with a deduced molecular weight of 32.15 kDa. Phylogenetic tree analysis suggested that DcTm1-X1 shares a high amino acid identity with its homolog in Acyrthosiphon pisum. Higher DcTm1-X1 expression levels were found in the leg of the psyllid by reverse transcription quantitative PCR (RT-qPCR). According to Blue Native PAGE analysis and mass spectrometric analysis, DcTm1-X1 interacts with citrate synthase (CS) and V-type proton ATPase subunit B-like (VAT). In addition, knockdown of DcTm1-X1 by RNA interference (RNAi) significantly increased the mortality rate of nymphs and the infection rate of CLas at different time points. Taken together, our results show that DcTm1-X1 might play an important role in response to CLas, but also lay a foundation for further research on the functions of DcTm1-X1.


Subject(s)
Hemiptera/metabolism , Insect Vectors/metabolism , Tropomyosin/metabolism , Amino Acid Sequence , Animals , Base Sequence , Gene Expression , Hemiptera/genetics , Hemiptera/microbiology , Insect Proteins/genetics , Insect Proteins/metabolism , Insect Vectors/genetics , Plant Diseases , Protein Isoforms/metabolism , Recombinant Proteins/isolation & purification , Tropomyosin/genetics
12.
Gynecol Endocrinol ; 34(9): 784-788, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29648472

ABSTRACT

Polycystic ovary syndrome (PCOS) is a common endocrine disease that affects reproductive-aged women and mostly characterized by insulin resistance (IR). The underlying mechanism remains unknown. Long noncoding RNAs (lncRNAs) have been demonstrated to be involved in various levels of biological regulation process of cell development, metabolism, and differentiation. This study aims to investigate the relationship between IR and differential expression of lncRNA Growth-arrest specific transcript 5 (GAS5) in patients' serum with and without PCOS. A total of 76 cases of serum was collected from non-PCOS and PCOS patients with and without IR to measure interleukin-18 (IL-18) and GAS5 expression, which were correlated with IR status. The IL-18 concentration in serums was significantly increased in PCOS patients with IR. GAS5 expression was decreased in serums in PCOS patients with IR. Result of correlation analysis shows that there is a negative association between GAS5 expression and homeostasis model of assessment for insulin resistance (HOMA-IR). GAS5 was yielded the ROC curve (AUC). Our study implied that elevated IL-18 expression and downregulation of GAS5 in serums might contribute to IR in PCOS patients.


Subject(s)
Down-Regulation , Insulin Resistance/physiology , Polycystic Ovary Syndrome/metabolism , RNA, Long Noncoding/blood , Adult , Anti-Mullerian Hormone/blood , Estradiol/blood , Female , Follicle Stimulating Hormone/blood , Humans , Interleukin-18/blood , Luteinizing Hormone/blood , Polycystic Ovary Syndrome/blood , Testosterone/blood , Young Adult
13.
J Assist Reprod Genet ; 35(1): 177-186, 2018 Jan.
Article in English | MEDLINE | ID: mdl-28921398

ABSTRACT

PURPOSE: The aim of this study was to determine whether an interchromosomal effect (ICE) occurred in embryos obtained from reciprocal translocation (rcp) and Robertsonian translocation (RT) carriers who were following a preimplantation genetic diagnosis (PGD) with whole chromosome screening with an aCGH and SNP microarray. We also analyzed the chromosomal numerical abnormalities in embryos with aneuploidy in parental chromosomes that were not involved with a translocation and balanced in involved parental translocation chromosomes. METHODS: This retrospective study included 832 embryos obtained from rcp carriers and 382 embryos from RT carriers that were biopsied in 139 PGD cycles. The control group involved embryos obtained from age-matched patient karyotypes who were undergoing preimplantation genetic screening (PGS) with non-translocation, and 579 embryos were analyzed in the control group. A single blastomere at the cleavage stage or trophectoderm from a blastocyst was biopsied, and 24-chromosomal analysis with an aCGH/SNP microarray was conducted using the PGD/PGS protocols. Statistical analyses were implemented on the incidences of cumulative aneuploidy rates between the translocation carriers and the control group. RESULTS: Reliable results were obtained from 138 couples, among whom only one patient was a balanced rcp or RT translocation carrier, undergoing PGD testing in our center from January 2012 to June 2014. For day 3 embryos, the aneuploidy rates were 50.7% for rcp carriers and 49.1% for RT carriers, compared with the control group, with 44.8% at a maternal age < 36 years. When the maternal age was ≥ 36 years, the aneuploidy rates were increased to 61.1% for rcp carriers, 56.7% for RT carriers, and 60.3% for the control group. There were no significant differences. In day 5 embryos, the aneuploidy rates were 24.5% for rcp carriers and 34.9% for RT carriers, compared with the control group with 53.6% at a maternal age < 36 years. When the maternal age was ≥ 36 years, the aneuploidy rates were 10.7% for rcp carriers, 26.3% for RT carriers, and 57.1% for the control group. The cumulative aneuploidy rates of chromosome translocation carriers were significantly lower than the control group. No ICE was observed in cleavage and blastocyst stage embryos obtained from these carriers. Additionally, the risk of chromosomal numerical abnormalities was observed in each of the 23 pairs of autosomes or sex chromosomes from day 3 and day 5 embryos. CONCLUSION: There was not enough evidence to prove that ICE was present in embryos derived from both rcp and RT translocation carriers, regardless of the maternal age. However, chromosomal numerical abnormalities were noticed in 23 pairs of autosomes and sex chromosomes in parental structurally normal chromosomes. Thus, 24-chromosomal analysis with an aCGH/SNP microarray PGD protocol is required to decrease the risks of failure to diagnose aneuploidy in structurally normal chromosomes.


Subject(s)
Chromosome Aberrations , DNA Copy Number Variations , Preimplantation Diagnosis/methods , Translocation, Genetic , Adult , Case-Control Studies , Comparative Genomic Hybridization , Female , Fertilization in Vitro , High-Throughput Nucleotide Sequencing , Humans , Microarray Analysis , Pilot Projects , Polymorphism, Single Nucleotide , Pregnancy , Pregnancy Rate , Retrospective Studies
14.
J Nanosci Nanotechnol ; 15(2): 1135-42, 2015 Feb.
Article in English | MEDLINE | ID: mdl-26353623

ABSTRACT

In an effort to examine liver, heart and kidney injury, immune response, and other physiological effect in rats caused by intratracheal instillation of nano titanium dioxide (TiO2) for 28 days, we assessed T lymphocytes counts, hematological indices, biochemical parameters, cytokines assay and histopathological changes in nano TiO2 treated rats. Indeed, rats treated with nano TiO2 displayed a reduction in body weight and coefficients of the hearts. Edema and loose cytoplasm on liver cells were found in nano groups. The results showed that a statistically significant increased in the BUN, HTC and AST levels than those in control group. Our data suggested that the immunologically competent cells of CD3+, CD4+, and CD8+ caused by nano TiO2 32 mg/kg group were significantly lower than control group. The ratio of CD4+ to CD8+ from the nano 32 mg/kg group was significantly increased and showed a disturbance of cellular immune function. But ELISA analysis showed that no significant changes in IFN-γ and IL-4 were observed throughout the experimental period in this study.


Subject(s)
Cytokines/immunology , Metal Nanoparticles/toxicity , Metal Nanoparticles/ultrastructure , Systemic Inflammatory Response Syndrome/chemically induced , Systemic Inflammatory Response Syndrome/immunology , Titanium/toxicity , Animals , Immunity, Innate/drug effects , Immunity, Innate/immunology , Materials Testing , Metal Nanoparticles/chemistry , Particle Size , Rats , Titanium/chemistry
15.
Int J Mol Med ; 35(3): 569-78, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25524499

ABSTRACT

The present study aimed to investigate the X chromosome inactivation (XCI) status in long-term cultured human parthenogenetic embryonic stem cells. One human embryonic stem (hES) cell line and 2 human parthenogenetic embryonic stem (hPES) cell lines were subjected to long-term culture in vitro (>50 passages). Karyotyping, array-based comparative genomic hybridization (aCGH), X-inactive specific transcript (XIST) RNA, immunofluorescence staining and real-time PCR were used to assess the chromosome karyotypes of these cells and the XCI status. X chromosome microdeletion was observed in the hPES-2 cells following culture for 50 passages. As early as 20 passages, XIST RNA expression was detected in the hPES-2 cells and was followed by low X-linked gene expression. The XIST RNA expression level was higher in the differentiated hPES-2 cells. The hPES-2' cells that were subclones of hPES-2 retained the XCI status, and had low XIST and X-linked gene expression. XIST RNA expression remained at a low level in the differentiated hPES-2' cells. The human biparental embryonic stem (hBES)-1 and hPES-1 cells did not exhibit XCI, and the differentiated hPES-1 cells had high expression levels of XIST RNA. In conclusion, the chromosome karyotypes of some hPES cell lines revealed instabilities. Similar to the hES cells, the hPES cells exhibited 3 XCI statuses. The unstable XCI status of the hPES-2 line may have been related to chromosome instability. These unstable chromosomes renedered these cells susceptible to environmental conditions and freezing processes, which may be the result of environmental adaptations.


Subject(s)
Embryonic Stem Cells/metabolism , Parthenogenesis/genetics , X Chromosome Inactivation , Cell Culture Techniques , Cell Line , Cells, Cultured , Gene Expression Regulation, Developmental , Genes, X-Linked , Genomic Instability , Histones/metabolism , Humans , Karyotype , RNA, Long Noncoding/genetics
16.
Zhonghua Fu Chan Ke Za Zhi ; 49(3): 193-8, 2014 Mar.
Article in Chinese | MEDLINE | ID: mdl-24820304

ABSTRACT

OBJECTIVE: To investigate the clinical use of array comparative genomic hybridization (aCGH) with fluorescence in situ hybridization (FISH) in preimplantion genetic diagnosis (PGD) for reciprocal and Robertsonian translocation carriers. METHODS: From Jan. 2012 to Jun. 2013, a total of 220 PGD cycles from 151 reciprocal translocation and 62 Robertsonian translocation carrier couples, including 33 cycles for reciprocal translocation carriers and 22 cycles for Robertsonian translocation carriers performed using array CGH, and 119 cycles for reciprocal translocation carriers and 46 cycles for Robertsonian translocation carriers performed using FISH were retrospectively studied. The rate of accurate diagnosis was compared between two methods. RESULTS: Normal and/or balance rates of the two translocated chromosomes detected by aCGH for both reciprocal and Robertsonian translocation carriers were 38.20% (123/322) and 67.20% (127/189), significantly higher than 15.39% (195/1 267) and 30.75% (202/657) by FISH (all P < 0.05). Abnormal rates of the two translocated chromosomes detected by aCGH for both reciprocal and Robertsonian translocation carriers were 59.32% (191/322) and 30.69% (58/189), significantly lower than 83.03% (1 052/1 267) and 67.43% (443/657) by FISH (all P < 0.05). And the rate of aneu ploidy in non-translocated chromosome from reciprocal translocation embryos was 20.19% (65/322), which was significantly lower than 38.62% (13/189) from Robertsonian translocation embryos (P < 0.01). CONCLUSIONS: Normal and/or balance rates of the two translocated chromosomes detected by array CGH were significantly higher than FISH. And the rate of aneuploidy in non-translocated chromosomes from reciprocal translocation embryos was significantly lower than that from Robertsonian translocation embryos.


Subject(s)
Chromosome Disorders/diagnosis , Comparative Genomic Hybridization , In Situ Hybridization, Fluorescence , Preimplantation Diagnosis/methods , Translocation, Genetic , Adult , Aneuploidy , Biopsy , Blastocyst , Embryo Culture Techniques , Embryo Transfer , Female , Humans , Pregnancy , Pregnancy Outcome
17.
Microb Pathog ; 63: 54-8, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23823084

ABSTRACT

Mycoplasmas are widely distributed among animals, plants, and human. The four species namely, Mycoplasmas genitalium(Mg), Mycoplasmas fermentans(Mf), Mycoplasmas pentrans(Mpe), Mycoplasmas pirum(Mpi) are also called AIDS-associated mycoplasmas due to their involvement in the development and outcome of AIDS. To investigate the infection prevalence of Mg, Mf, Mpe and Mpi among male HIV/AIDS patients in Jiangsu Province and to analyze the relationship between pathogenic mycoplasmas and cellular immune function of them. First void urine and venous blood samples were collected and epidemiology questionnaires were administered after informed consent. Nested PCR was performed to determine the infection of Mg, Mf, Mpe and Mpi while ELISA assay was applied to detect interleukin-2(IL-2), interferon-γ(IFN-γ) and tumor necrosis factor-α(TNF-α). SAS 9.0 software was applied to analyze the data. A total of 713 HIV/AIDS patients were recruited in this study. The overall infection rates of Mg, Mf, Mpe and Mpi are 27.9%, 9.7%, 1.0% and 18.4% respectively. Generally, the infection rates of Mg(χ(2) = 10.311, P = 0.006) and Mpi were declined as the CD4+ cell counts increased, while Mf infection was higher in CD4+ T cell>350/µl group. The levels of cytokines are different with the variance of mycoplasmas infection. Mycoplasma infection among male HIV/AIDS patients is associated with changes in cellular immune response (cytokines). However, the affect of mycoplasmas on the immune function is complex, further studies are still required to elucidate whether mycoplasmas interact with HIV by interfering host immune system.


Subject(s)
HIV Infections/complications , Mycoplasma Infections/epidemiology , Mycoplasma/isolation & purification , Blood/microbiology , CD4 Lymphocyte Count , China , Cytokines/metabolism , HIV Infections/immunology , Humans , Male , Prevalence , Surveys and Questionnaires , Urine/microbiology
18.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 30(3): 283-7, 2013 Jun.
Article in Chinese | MEDLINE | ID: mdl-23744315

ABSTRACT

OBJECTIVE: To assess the value of array comparative genomic hybridization (array CGH) technique for preimplantation genetic diagnosis (PGD). METHODS: Array CGH was performed on three types of cells, which included 3-5 cells isolated from B2/C38/A1 embryonic stem cell lines, single cells isolated from two discarded normal fertilized embryos, and 10 blastocysts biopsied from 5 couples undergoing PGD for chromosomal translocations. For the 10 blastocysts, 8 were abnormal embryos, 1 appeared to be normal but showed arrested development, and 1 embryo was without any fluorescence signals. 24sure V3 or 24sure+ array chips were applied for CGH analysis. The results were analyzed with a BlueFuse Multi software. RESULTS: (1) The results of cells from B2/C3/A1 embryo stem cells by array CGH were consistent with karyotyping analysis. (2) For the 6 single cell samples from two discarded embryos, 2 blastomeres from one embryo were diagnosed as with aneuploidy and a normal karyotype, respectively. Two out of 4 blastomeres biopsied from another embryo were normal, whilst the remaining two were diagnosed with aneuploidies of -22 and +13. Repeated detection with 24sure+ array was consistent with the 24sure V3 result. (3) Ten cell masses from 10 embryos in PGD cycles were successfully analyzed with array CGH, among which four were confirmed with fluorescence in situ hybridization (FISH) on day 3. In two of them, array CGH confirmed FISH diagnosis. For the remaining two, additional aneuploidies for chromosomes not tested by FISH were discovered by array CGH. Another embryo diagnosed as no signal by FISH was found to have trisomy 13 by array CGH. The remaining 5 embryos also showed discordant results by FISH and array CGH. One embryo from a Robertsonian translocation carrier was found to have monosomy 13 by FISH but trisomy 14 and additional aneuploidies by both 24sure V3 and 24sure+ chips. One embryo with many fragments and arrested development by D5 showed discordant results by FISH and array CGH. However, the FISH and array CGH results for other two embryos from this reciprocal translocation carrier were consistent. Three embryos with inconsistent results by FISH and array CGH had a chromosomal translocation involving q11 region. CONCLUSION: Array CGH is useful for PGD applications for its capability to detect structural chromosomal abnormalities through screening of aneuploidies. However, the 24sure V3 array may not suit detection of translocations with breakpoints close to the q11 region of chromosomes.


Subject(s)
Comparative Genomic Hybridization , Preimplantation Diagnosis , Cell Line , Embryonic Stem Cells/metabolism , Female , Humans , In Situ Hybridization, Fluorescence , Male , Pregnancy , Reproducibility of Results , Translocation, Genetic
19.
Zhonghua Liu Xing Bing Xue Za Zhi ; 34(3): 259-62, 2013 Mar.
Article in Chinese | MEDLINE | ID: mdl-23759233

ABSTRACT

OBJECTIVE: To investigate the prevalence of Mycoplasma pirum (Mpi) in male HIV infected patients, and to identify the 16S rRNA gene of Mpi. METHODS: The first void urine of male HIV/AIDS patients in Jiangsu province was collected for Mpi detection. Purified 16S rRNA gene PCR production was sequenced for analysis on its identification, homogeneity and phylogenetic tree. P1 protein sequence of Mpi was analyzed by Vector NTI Advance 11.0 to calculate the coded amino acid sequence. Homogeneity analysis was conducted between the theoretical amino acid sequence of Mpi and other Mycoplasmas. RESULTS: The prevalence of Mpi in male HIV/AIDS patients was 21.5% while the Mpi prevalence rates in different age groups were significantly different (χ² Mpi = 124.63, P < 0.01). The homogeneity of 18 strains of Mpi was higher than 90%. CONCLUSION: The Mpi prevalence seemed much higher than the results from previous detection on HIV/AIDS patients, suggesting that more attention should be paid on AIDS treatment. More bioinformatic research on gene/nucleotide sequence analysis and forecast should be carried out to identify the molecular characteristics of Mpi.


Subject(s)
Acquired Immunodeficiency Syndrome/microbiology , HIV Infections/microbiology , Mycoplasma/genetics , RNA, Ribosomal, 16S , Acquired Immunodeficiency Syndrome/epidemiology , Adolescent , Adult , Aged , Child , Genes, rRNA , HIV Infections/epidemiology , Humans , Male , Middle Aged , Mycoplasma/classification , Mycoplasma Infections/epidemiology , RNA, Ribosomal, 16S/genetics , Young Adult
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