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1.
Yi Chuan ; 45(1): 88-95, 2023 Jan 20.
Article in English | MEDLINE | ID: mdl-36927641

ABSTRACT

Duchenne/Becker muscular dystrophy (DMD/BMD) is one of the most common progressive muscular dystrophy diseases with X-linked recessive inheritance. It is mainly caused by the deletion, duplication and point mutation of DMD gene. In rare cases, it is also caused by the destruction of DMD gene by chromosomal structural rearrangement. Here, we report a case of Duchenne/Becker Muscular dystrophy (DMD/BMD) with typical symptoms but unknown genetic defects after MLPA and next generation sequencing tests in other hospitals. Interestingly, we find a pericentric inversion of X chromosome (Chr.X: g. [31939463-31939465del; 31939466-131765063 inv; 131765064-131765067del]) in this patient. We then use the karyotyping, FISH, long-read sequencing and Sanger sequencing technologies to characterize the chromosome rearrangement. We find that this chromosomal aberration disrupt both the DMD gene and the HS6ST2 gene. The patient present with typical DMD symptoms such as muscle weakness, but no obvious symptoms of Paganini-Miozzo syndrome. Our results suggest that the destruction of DMD gene by structural rearrangement is also one of the important causes of DMD. Therefore, we suggest to provide further genetic testing for those DMD patients with unknown genetic defects through routine genetic testing. Cost-effective karyotyping and FISH should be considered firstly to identify chromosome rearrangements. Long-read sequencing followed by Sanger sequencing could be useful to locate the precise breakpoints. The genetic diagnosis of this case made it possible for reproductive intervention in the patient's family.


Subject(s)
Muscular Dystrophy, Duchenne , Humans , Muscular Dystrophy, Duchenne/genetics , Muscular Dystrophy, Duchenne/diagnosis , Dystrophin/genetics , Genetic Testing , Gene Rearrangement/genetics , X Chromosome , Sulfotransferases/genetics
2.
Drug Des Devel Ther ; 16: 2365-2382, 2022.
Article in English | MEDLINE | ID: mdl-35910781

ABSTRACT

Background: As the main component of turmeric (Curcuma longa L.), curcumin is widely used in the treatment of various diseases. Previous studies have demonstrated that curcumin has great potential as a therapeutic agent, but the lack of understanding of the functional mechanism of the drug has hindered the widespread use of the natural product. In the present study, we used comprehensive bioinformatics analysis and in vitro experiments to explore the anti-tumor mechanism of curcumin. Materials and Methods: LUAD mRNA expression data were obtained from TCGA database and differentially expressed genes (DEGs) were identified using R software. Functional enrichment analysis was conducted to further clarify its biological properties and hub genes were identified by a protein-protein interaction (PPI) network analysis. Survival analysis and molecular docking were used to analyze the effectiveness of the hub genes. By an in vitro study, we evaluated whether curcumin could influence the proliferation, migration, and invasion activities of LUAD cells. Results: In this study, 1783 DEGs from LUAD tissue samples compared to normal samples were evaluated. Functional enrichment analysis and the PPI network revealed the characteristics of the DEGs. We performed a topological analysis and identified 10 hub genes. Of these, six genes (INS, GCG, SST, F2, AHSG, and NPY) were identified as potentially effective biomarkers of LUAD. The molecular docking results indicated that curcumin targets in regulating lung cancer may be INS and GCG. We found that curcumin significantly inhibited the proliferation, migration, and invasion of LUAD cells and significantly decreased the expression of the INS and GCG genes. Conclusion: The results of this study suggest that the therapeutic effects of curcumin on LUAD may be achieved through the intervention of INS and GCG, which may act as potential biomarkers for LUAD prevention and treatment.


Subject(s)
Adenocarcinoma of Lung , Curcumin , Lung Neoplasms , Adenocarcinoma of Lung/drug therapy , Adenocarcinoma of Lung/genetics , Biomarkers, Tumor , Computational Biology , Curcumin/pharmacology , Gene Expression Regulation, Neoplastic , Humans , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Molecular Docking Simulation
3.
J Med Genet ; 59(10): 1010-1016, 2022 10.
Article in English | MEDLINE | ID: mdl-35121647

ABSTRACT

BACKGROUND: Numerous variants of uncertain significance (VUSs) have been identified by whole exome sequencing in clinical practice. However, VUSs are not currently considered medically actionable. OBJECTIVE: To assess the splicing patterns of 49 VUSs in 48 families identified clinically to improve genetic counselling and family planning. METHODS: Forty-nine participants with 49 VUSs were recruited from the Reproductive and Genetic Hospital of CITIC-Xiangya. Bioinformatic analysis was performed to preliminarily predict the splicing effects of these VUSs. RT-PCR and minigene analysis were used to assess the splicing patterns of the VUSs. According to the results obtained, couples opted for different methods of reproductive interventions to conceive a child, including prenatal diagnosis and preimplantation genetic testing (PGT). RESULTS: Eleven variants were found to alter pre-mRNA splicing and one variant caused nonsense-mediated mRNA decay, which resulted in the reclassification of these VUSs as likely pathogenic. One couple chose to undergo in vitro fertilisation with PGT treatment; a healthy embryo was transferred and the pregnancy is ongoing. Three couples opted for natural pregnancy with prenatal diagnosis. One couple terminated the pregnancy because the fetus was affected by short-rib thoracic dysplasia and harboured the related variant. The infants of the other two couples were born and were healthy at their last recorded follow-up. CONCLUSION: RNA splicing analysis is an important method to assess the impact of sequence variants on splicing in clinical practice and can contribute to the reclassification of a significant proportion of VUSs. RNA splicing analysis should be considered for genetic disease diagnostics.


Subject(s)
RNA Precursors , RNA Splicing , Female , Genetic Counseling , Genetic Testing/methods , Humans , Pregnancy , Prenatal Diagnosis , RNA Splicing/genetics
4.
Biomed Pharmacother ; 148: 112675, 2022 Apr.
Article in English | MEDLINE | ID: mdl-35183993

ABSTRACT

Acute graft-versus-host disease (aGVHD) is a major life-threatening complication after Allogeneic Hematopoietic Stem Cell Transplant (allo-HSCT). Although a series of immunosuppressant agents are routinely used as the first-line prevention, the morbidity and mortality rate remains high in allo-HSCT recipients. Our previous work indicated that combining Xuebijing (XBJ) with Cyclosporin A (CSA) is superior to CSA alone in preventing aGVHD. However, it was not clear which compounds in XBJ may prevent aGVHD. Whether the effective compounds in XBJ can be safely combined with CSA to prevent GVHD remain to be evaluated. Here, we accessed whether the combination of four main components in XBJ (C0127) had the same efficacy as XBJ in preventing aGVHD. In addition, the effectiveness of a novel combination therapy (C0127 + CSA) on aGVHD prophylaxis was evaluated using 16 s rRNA sequencing and RNA sequencing approaches in vitro and in vivo. In aGVHD mice, C0127 enhanced the preventive effects of CSA including decreasing mortality, maintaining weight, reducing GVHD score and reducing the expression of IL-6 and TNF-α in serum. Fatal GVHD is a frequent consequence of intestinal tract damage. We found combining C0127 with CSA alleviated the gut damage and maintained the normal physiological function of intestine by H&E staining, intestinal permeability and short chain fatty acid (SCFA) assays. Next, 16 S sequencing analysis of feces showed the combination treatment maintained the intestinal microbial diversity, normalized the intestinal microorganism and prevented flora disorder by reducing the relative abundances of Escherichia coli and Enterococcus. Further, RNA-seq analysis of colonic epithelium revealed C0127 combined with CSA chiefly regulated chemokines and cytokines in IL-17 signaling pathway. The combination treatment reduced the expression of G-CSF and its effector STAT3 (an axis that aggravated gut inflammation and flora disorder) in gut epithelium on mRNA and protein level. These findings indicated that C0127 improved the prevention of CSA in aGVHD mice partially by protecting the gut from damage through normalizing G-CSF signaling, which regulates the intestinal microbiota and the integrity of the epithelial barrier.


Subject(s)
Drugs, Chinese Herbal , Graft vs Host Disease , Hematopoietic Stem Cell Transplantation , Acute Disease , Animals , Cyclosporine/pharmacology , Drugs, Chinese Herbal/pharmacology , Drugs, Chinese Herbal/therapeutic use , Graft vs Host Disease/prevention & control , Hematopoietic Stem Cell Transplantation/adverse effects , Mice
5.
Acta Trop ; 224: 106073, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34487719

ABSTRACT

In western societies, the prevalence of type 2 diabetes (T2D) is related to the hygiene hypothesis, which implies that reduced exposure to infectious factors results in a loss of the immune stimulation necessary to form the immune system during development. In fact, it has been reported that parasites, such as Schistosoma, can improve or prevent the development of T2D, which may be related to the activity of immune cells, including regulatory T cells (Tregs). Hence, Schistosoma, Tregs, and T2D share a close relationship. Schistosoma infection and the molecules released can lead to an increase in Tregs, which play an important role in the suppression of T2D. In this review, we provide an overview of the role of Tregs in the response to Schistosoma infection and the protective mechanism of Schistosoma-related molecular products against T2D.


Subject(s)
Diabetes Mellitus, Type 2 , T-Lymphocytes, Regulatory , Animals , Diabetes Mellitus, Type 2/prevention & control , Schistosoma
6.
Cancer Manag Res ; 13: 4935-4946, 2021.
Article in English | MEDLINE | ID: mdl-34188546

ABSTRACT

BACKGROUND: There are no useful biomarkers for the clinical outcome of advanced esophageal squamous cell carcinoma (ESCC). In this study, we aimed to investigate the prognostic value of soluble PD-L1 (sPD-L1) in serum of patients with locally advanced or metastatic ESCC who received cytotoxic chemotherapy as first-line treatment. MATERIALS AND METHODS: This study evaluated the expression pattern of PD-L1 by immunohistochemistry and sPD-L1 concentration, and correlation with clinicopathological factors and overall survival (OS) in 190 patients with ESCC. RESULTS: sPD-L1 concentration was highly expressed in ESCC, especially in female patients. Patients with a high sPD-L1 level (≥0.63 ng/mL) had a shorter OS than those with a low sPD-L1 level (<0.63 ng/mL). In a multivariate analysis, high sPD-L1 concentration remained an independent prognostic factor of OS after adjustment for possible confounders. However, tissue PD-L1 expression level was non-prognostic in this study. CONCLUSION: There was no significant correlation between serum sPD-L1 concentration and tissue PD-L1 expression level. sPD-L1 concentration before treatment could be an effective and convenient biomarker of prognosis in patients with locally advanced or metastatic ESCC treated with combination cytotoxic chemotherapy.

7.
J Parasitol ; 107(3): 472-480, 2021 05 01.
Article in English | MEDLINE | ID: mdl-34153095

ABSTRACT

We investigated the effect of Schistosoma japonicum adenylate kinase 1 (Sjak1) on the growth and development of schistosomula. Quantitative real-time PCR showed that Sjak1 mRNA was expressed in 3-, 10-, 14-, 18-, and 21-day-old schistosomula, and its levels increased gradually with the development of S. japonicum. Using immunohistochemical techniques, ak1 protein was found to be mainly distributed in the tegument and some parenchymal tissues of the schistosomula. Double-stranded RNA-mediated knockdowns of ak1 decreased ak1 mRNA transcripts by more than 90%, and western blot results showed that expression of ak1 protein was decreased by 66%. Scanning electron microscopy following the RNA-mediated ak1 knockdown showed that the sensory papillae did not develop. Transmission electron microscopy showed a lower mean thickness of the tegument in the Sjak1 interference group than in the negative control group. Terminal deoxynucleotidyl transferase dUTP nick-end labeling suggested higher apoptosis in the interference group than the negative control group. These results showed that ak1 may be involved in the growth and development of S. japonicum schistosomula and especially in the development of the integument. Consequently, ak1 may be a potential target in developing prevention methods for schistosomiasis in the future.


Subject(s)
Adenylate Kinase/metabolism , Schistosoma japonicum/enzymology , Schistosoma japonicum/growth & development , Adenylate Kinase/analysis , Adenylate Kinase/genetics , Animals , Apoptosis , Blotting, Western , DNA/physiology , Female , Gene Expression Regulation, Enzymologic , Gene Knockdown Techniques/methods , Gene Silencing , Immunohistochemistry , In Situ Nick-End Labeling , Liver/parasitology , Mice , Mice, Inbred ICR , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , RNA Interference , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rabbits , Real-Time Polymerase Chain Reaction , Schistosoma japonicum/genetics , Schistosoma japonicum/ultrastructure , Snails/parasitology
8.
Gene ; 788: 145666, 2021 Jul 01.
Article in English | MEDLINE | ID: mdl-33887368

ABSTRACT

BACKGROUND: Recent studies in cancer biology suggest that metabolic glucose reprogramming is a potential target for cancer treatment. However, little is known about drug intervention in the glucose metabolism of cancer stem cells (CSCs) and its related underlying mechanisms. METHODS: The crude realgar powder was Nano-grinded to meets the requirements of Nano-pharmaceutical preparations, and Nano-realgar solution (NRS) was prepared for subsequent experiments. Isolation and characterization of lung cancer stem cells (LCSCs) was performed by magnetic cell sorting (MACS) and immunocytochemistry, respectively. Cell viability and intracellular glucose concentration were detected by MTT assay and glucose oxidase (GOD) kit. Protein expressions related to metabolic reprogramming was detected by ELISA assay. Determination of the expression of HIF-1α and PI3K/Akt/mTOR pathways was carried out by RT-PCR and western blotting analysis. A subcutaneous tumor model in BALB/c-nu mice was successfully established to evaluate the effects of Nano-realgar on tumor growth and histological structure, and the expression of HIF-1α in tumor tissues was measured by immunofluorescence. RESULTS: Nano-realgar inhibits cell viability and induces glucose metabolism in LCSCs, and inhibits protein expression related to metabolic reprogramming in a time- and dose-dependent manner. Nano-realgar downregulated the expression of HIF-1α and PI3K/Akt/mTOR pathways in vitro and in vivo. Nano-realgar inhibits tumor growth and changes the histological structure of tumors through in vivo experiments and consequently inhibits the constitutive activation of HIF-1α signaling. CONCLUSIONS: These results reveal that Nano-realgar inhibits tumor growth in vitro and in vivo by repressing metabolic reprogramming. This inhibitory effect potentially related to the downregulation HIF-1α expression via PI3K/Akt/mTOR pathway.


Subject(s)
Antineoplastic Agents/administration & dosage , Arsenicals/administration & dosage , Glucose/metabolism , Lung Neoplasms/drug therapy , Neoplastic Stem Cells/metabolism , Sulfides/administration & dosage , A549 Cells , AC133 Antigen/metabolism , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Arsenicals/chemistry , Arsenicals/pharmacology , Cell Line , Cell Proliferation/drug effects , Cell Survival/drug effects , Gene Expression Regulation, Neoplastic/drug effects , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Lung Neoplasms/metabolism , Male , Mice , Mice, Inbred BALB C , Nanoparticles , Neoplastic Stem Cells/drug effects , Phosphatidylinositol 3-Kinases/genetics , Phosphatidylinositol 3-Kinases/metabolism , Sulfides/chemistry , Sulfides/pharmacology , TOR Serine-Threonine Kinases/genetics , TOR Serine-Threonine Kinases/metabolism , Xenograft Model Antitumor Assays
9.
Infect Genet Evol ; 85: 104481, 2020 11.
Article in English | MEDLINE | ID: mdl-32736043

ABSTRACT

Taenia hydatigena is a cestode of veterinary importance. Infection with the metacestode larval stage results in cysticercosis, which poses a serious challenge to the livestock industry worldwide. Globally, there are numerous reports on cysticercosis caused by T. hydatigena in sheep and goat but a lack of data on the prevalence and genetic diversity exists for Pakistan. We designed this study to provide an insight into the disease status as well as investigate the genetic variation among the recovered isolates based on the mitochondrial cox1 gene. In this study, we examined small ruminants (sheep and goats) slaughtered in Faisalabad in eastern Punjab province of Pakistan for T. hydatigena metacestodes and described the population structure and genetic variation using the cytochrome c oxidase subunit 1 (cox1) mitochondrial gene. Overall, a prevalence of 4.40% (goat =4.67% sheep = 4.07%) from a total of 2225 small ruminant carcasses (sheep = 983, goats = 1242) was observed. Based on the NCBI BLAST search and Bayesian phylogeny, the identity of all isolates was confirmed via their nucleotide sequences. The diversity indices indicated a high haplotype and a low nucleotide diversity with 43 haplotypes from 98 isolates. The results also show the existence of unique haplotypes of T. hydatigena in Pakistan as demonstrated by the significant negative values of Tajima's D and Fu's Fs neutrality test suggesting a recent population expansion. The median-joining network of the partial cox1 sequence dataset showed the existence of two main haplotypes detected in both sheep and goat populations. This study shows that the prevalence of cycticercosis due to T. hydatigena is below 5% in sheep and goats in Faisalabad, Punjab, Pakistan. The molecular analysis of the partial cox1 gene also indicates a high degree of genetic variation with the existence of rare haplotypes. These findings represent a preliminary report on the prevalence and genetic variation of T. hydatigena in Pakistan and serve as baseline information for future studies on the prevalence and population structure of T. hydatigena in the country.


Subject(s)
Cyclooxygenase 1/genetics , Cysticercosis/parasitology , Genes, Mitochondrial , Goats/parasitology , Haplotypes , Sheep/parasitology , Taenia/genetics , Animals , Bayes Theorem , Cysticercosis/epidemiology , Genetic Variation , Goat Diseases/epidemiology , Goat Diseases/parasitology , Pakistan/epidemiology , Phylogeny , Prevalence , Serogroup , Sheep Diseases/epidemiology , Sheep Diseases/parasitology
10.
Shanghai Kou Qiang Yi Xue ; 29(6): 586-590, 2020 Dec.
Article in Chinese | MEDLINE | ID: mdl-33778823

ABSTRACT

PURPOSE: To investigate the effect of firing times on the fatigue properties and failure modes of posterior occlusal veneers made of lithium disilicate glass ceramics. METHODS: According to the number of times of firing (1, 3, 5, 7 times), IPS e.max CAD(IC) and IPS e.max Press(IP) occlusal veneer restorations with a thickness of 1.2mm were prepared, and then cemented to the maxillary molar composite resin dies. Each group was subjected to thermo-mechanical fatigue(TMFT) test (5-55 ℃, 5000 cycles, 30-300 N, 10 Hz, sinusoidal wave, 500 000 cycles) and compressive loading. The maximum force at fracture was recorded. Stereomicroscope and scanning electron microscope(SEM), energy dispersive X-ray spectroscopy(EDX) and X-ray diffraction(XRD) were adopted to analyze the failure modes and microstructural features. Statistical analysis was performed with SPSS 20.0 software package. RESULTS: The highest average loads (N) at fracture for IC groups were (1546.73±192.85) N after one firing time; in IP group, the highest average loads were (1504.46±138.56) N after three firing times. There was no significant difference in the fracture load with different firing times for IC/IP(P>0.05). EDX and XRD analysis showed no significant difference in the composition of two materials. SEM showed that the size of crystal was shorter and the porosity increased after multiple firing. CONCLUSIONS: After repeated firing, the fatigue loading of lithium disilicate glass ceramics has a decreasing trend of fracture load value without any significant difference, and it meets clinical requirements.


Subject(s)
Dental Porcelain , Dental Veneers , Ceramics , Composite Resins , Computer-Aided Design , Dental Restoration Failure , Dental Stress Analysis , Materials Testing , Surface Properties
11.
Acta Trop ; 202: 105239, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31669534

ABSTRACT

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a candidate subunit vaccine that induces protective immunity and elicits partial resistance to Schistosoma japonicum upon mouse and livestock vaccination. This study aimed to evaluate the effect of regulatory T cells (Tregs), which were defined as CD4+CD25+Foxp3+ cells, on the efficacy of a GAPDH vaccine against S. japonicum. BALB/c female mice were randomly divided into five groups as follows: normal, infected control, anti-CD25 monoclonal antibody (anti-CD25 mAb), GAPDH group, and co-treated with anti-CD25 mAb and GAPDH group. The worm reduction and liver egg reduction rates in the GAPDH group were 32.46% and 35.43%, respectively, which increased to 60.09% and 58.78%, respectively, after anti-CD25 mAb administration. Compared with those in the infected control group, the percentage of Tregs in the spleen decreased significantly when GAPDH and anti-CD25 mAb were used either alone or in combination. Furthermore, secretions associated with the Th1 response increased in splenocytes of the anti-CD25 mAb group, whereas the Th1 and Th2 responses increased in splenocytes of the GAPDH and co-treated groups. Compared to that in the infected control group, granuloma diameter in the GAPDH and co-treated groups increased slightly, but there were no significant differences among the groups. Our results indicate that the protective effect of the GAPDH vaccines can be improved by decreasing Tregs and enhancing the Th1- and Th2-type immune responses. Therefore, anti-CD25 mAb and GAPDH might exert synergistic effects to clear parasites by decreasing the frequency of Tregs and increasing the Th1- and Th2-type immune responses.


Subject(s)
Glyceraldehyde-3-Phosphate Dehydrogenases/immunology , Schistosoma japonicum/immunology , T-Lymphocytes, Regulatory/immunology , Vaccines/immunology , Animals , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/therapeutic use , Cytokines/immunology , Female , Granuloma/pathology , Interleukin-2 Receptor alpha Subunit/immunology , Liver/immunology , Liver/pathology , Mice , Mice, Inbred BALB C , Schistosomiasis japonica/parasitology , Schistosomiasis japonica/prevention & control , T-Lymphocytes, Regulatory/drug effects
12.
Biomed Res Int ; 2019: 2063823, 2019.
Article in English | MEDLINE | ID: mdl-31061821

ABSTRACT

BACKGROUND: Although several studies have proved the relationship between the prognostic value of miRNA-15a and different types of cancer, the result remains controversial. Thus, a meta-analysis was conducted to clarify the prognostic value of miRNA-15a expression level in human cancers. METHODS: We enrolled appropriate literature by searching the databases of PubMed, Embase, and Web of Science. Subsequently, we extracted HRs and their 95% CIs and calculated pooled results of miRNA-15a for overall survival (OS) and disease-free survival (DFS). Besides, subgroup analysis, sensitivity analysis, and publication bias were also revealed in this study. We also further validated this meta-analysis using the Kaplan-Meier plotter database. RESULT: 10 studies, including 1616 patients, were embraced in our meta-analysis. The result showed the lower expression of miRNA-15a significantly predicted adverse OS (HR=2.17, 95% CI: 1.41-3.34), but there is no significant association between the expressing level and DFS in cancer patient (HR=2.04, 95% CI: 0.60-6.88). Based on Kaplan-Meier plotter database, we found the same results in bladder Carcinoma, head-neck squamous cell carcinoma, liver hepatocellular carcinoma, lung squamous cell carcinoma, pancreatic ductal adenocarcinoma, rectum adenocarcinoma, stomach adenocarcinoma, and uterine corpus endometrial carcinoma, but opposite results were found in cervical squamous cell carcinoma and esophageal carcinoma. CONCLUSION: Low expressing levels of miRNA-15a indicated poor OS, while miRNA-15a can be used as a prediction biomarker in different cancer types.


Subject(s)
Biomarkers, Tumor/biosynthesis , Computational Biology , Gene Expression Regulation, Neoplastic , Neoplasms/metabolism , Neoplasms/mortality , RNA, Neoplasm/biosynthesis , Biomarkers, Tumor/genetics , Disease-Free Survival , Humans , MicroRNAs , Neoplasms/genetics , Neoplasms/pathology , Predictive Value of Tests , RNA, Neoplasm/genetics , Survival Rate
13.
BMC Med Genet ; 19(1): 186, 2018 10 17.
Article in English | MEDLINE | ID: mdl-30333007

ABSTRACT

BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD), the commonest inherited kidney disease, is generally caused by heterozygous mutations in PKD1, PKD2, or GANAB (PKD3). METHODS: We performed mutational analyses of PKD genes to identify causative mutations. A set of 90 unrelated families with ADPKD were subjected to mutational analyses of PKD genes. Genes were analysed using long-range PCR (LR-PCR), direct PCR sequencing, followed by multiplex ligation-dependent probe amplification (MLPA) or screening of GANAB for some patients. Semen quality was assessed for 46 male patients, and the correlation between mutations and male infertility was analysed. RESULTS: A total of 76 mutations, including 38 novel mutations, were identified in 77 families, comprising 72 mutations in PKD1 and 4 in PKD2, with a positive detection rate of 85.6%. No pathogenic mutations of GANAB were detected. Thirty-seven patients had low semen quality and were likely to be infertile. No association was detected between PKD1 mutation type and semen quality. However, male patients carrying a pathogenic mutation in the Ig-like repeat domain of PKD1 had a high risk of infertility. CONCLUSION: Our study identified a group of novel mutations in PKD genes, which enrich the PKD mutation spectrum and might help clinicians to make precise diagnoses, thereby allowing better family planning and genetic counselling. Men with ADPKD accompanied by infertility should consider intracytoplasmic sperm injection combined with preimplantation genetic diagnosis to achieve paternity and obtain healthy progeny.


Subject(s)
Genetic Predisposition to Disease , Infertility, Male/genetics , Mutation , Polycystic Kidney, Autosomal Dominant/genetics , TRPP Cation Channels/genetics , Adult , Asian People , DNA Mutational Analysis , Female , Gene Expression , Genetic Counseling , Glucosidases/genetics , Humans , Infertility, Male/diagnosis , Infertility, Male/ethnology , Infertility, Male/pathology , Male , Middle Aged , Multiplex Polymerase Chain Reaction , Patient Acceptance of Health Care , Polycystic Kidney, Autosomal Dominant/diagnosis , Polycystic Kidney, Autosomal Dominant/ethnology , Polycystic Kidney, Autosomal Dominant/pathology , Reproductive Techniques, Assisted , Semen Analysis
14.
Mol Genet Genomic Med ; 6(2): 261-267, 2018 03.
Article in English | MEDLINE | ID: mdl-29378385

ABSTRACT

BACKGROUND: Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a malformation of the eyelids. Forkhead Box L2 (FOXL2) is the only gene known to be associated with BPES. METHODS: We identified two Han Chinese BPES families with premature ovarian insufficiency (POI). Sanger sequencing and in vitro functional analysis were performed to identify the genetic cause. RESULTS: Sanger sequencing identified two novel mutations (c.462_468del, c.988_989insG) in FOXL2, one in each family. The in vitro functional analysis confirmed that both novel mutations were associated with impaired transactivation of downstream genes. Specifically, the single-base insertion, c.988_989insG, led to subcellular mislocalization and aggregation of the encoded protein, which validated the hypothesis that the two novel FOXL2 mutations are deleterious and associated with POI in the two BPES families. CONCLUSION: The novel mutations identified in the present study will enhance the present knowledge of the mutation spectrum of FOXL2. The in vitro experiments provide further insights into the molecular mechanism by which the two new variants mediate disease pathogenesis and may contribute to elucidating the genotype-phenotype correlation between the two novel FOXL2 mutations and POI.


Subject(s)
Blepharophimosis/genetics , Forkhead Box Protein L2/genetics , Primary Ovarian Insufficiency/genetics , Skin Abnormalities/genetics , Urogenital Abnormalities/genetics , Adult , Base Sequence/genetics , Blepharophimosis/complications , Blepharophimosis/metabolism , China , Ethnicity/genetics , Eyelids/abnormalities , Female , Forkhead Box Protein L2/metabolism , Forkhead Transcription Factors/genetics , Genetic Association Studies , Humans , Pedigree , Primary Ovarian Insufficiency/complications , Skin Abnormalities/metabolism , Urogenital Abnormalities/metabolism
15.
Zhongguo Dang Dai Er Ke Za Zhi ; 18(8): 746-50, 2016 Aug.
Article in Chinese | MEDLINE | ID: mdl-27530794

ABSTRACT

OBJECTIVE: To investigate the mutation of related genes and prenatal diagnosis of a family with Bartter syndrome (BS). METHODS: The high-throughput capture sequencing technique and PCR-Sanger sequencing were used to detect pathogenic genes in the proband of this family and analyze the whole family at the genomic level. After the genetic cause was clarified, the amniotic fluid was collected from the proband's mother who was pregnant for 5 months for prenatal diagnosis. RESULTS: The proband carried compound heterozygous mutations of c.88C>T(p.Arg30*) and c.968+2T>A in the CLCNKB gene; c.88C>T(p.Arg30*) had been reported as a pathogenic mutation, and c.968+2T>A was a new mutation. Pedigree analysis showed that the two mutations were inherited from the mother and father, respectively. Prenatal diagnosis showed that the fetus did not inherit the mutations from parents and had no mutations at the two loci. The follow-up visit confirmed that the infant was in a healthy state, which proved the accuracy of genetic diagnosis and prenatal diagnosis. CONCLUSIONS: The compound heterozygous mutations c.88C>T(p.Arg30*) and c.968+2T>A in the CLCNKB gene are the cause of BS in the proband, and prenatal diagnosis can prevent the risk of recurrence of BS in this family.


Subject(s)
Bartter Syndrome/genetics , Mutation , Prenatal Diagnosis , Bartter Syndrome/diagnosis , Female , Humans , Infant , Pregnancy
16.
Cell Physiol Biochem ; 36(5): 1688-98, 2015.
Article in English | MEDLINE | ID: mdl-26196155

ABSTRACT

BACKGROUND: Uncoupling protein 2 (UCP2) is critical in regulating energy metabolism. Due to the significant change in energy metabolism of myocardium upon pressure overload, we hypothesize that UCP2 could contribute to the etiology of cardiac hypertrophy. METHODS: Adult male C57BL/6J mice were subjected to pressure overload by using transverse aortic constriction (TAC), and then received genipin (a UCP2 selective inhibitor; 25 mg/kg/d, ip) or vehicle for three weeks prior to histologic assessment of myocardial hypertrophy. ATP concentration, ROS level, and myocardial apoptosis were also examined. A parallel set of experiments was also conducted in UCP2-/- mice. RESULTS: TAC induced left ventricular hypertrophy, as reflected by increased ventricular weight/thickness and increased size of myocardial cell (vs. sham controls). ATP concentration was decreased; ROS level was increased. Apoptosis and fibrosis markers were increased. TAC increased mitochondrial UCP2 expression in the myocardium at both mRNA and protein levels. Genipin treatment attenuated cardiac hypertrophy and the histologic/biochemical changes described above. Hypertrophy and associated changes induced by TAC in UCP2-/- mice were much less pronounced than in WT mice. CONCLUSIONS: Blocking UCP2 expression attenuates cardiac hypertrophy induced by pressure overload.


Subject(s)
Aorta/pathology , Cardiomegaly/prevention & control , Ion Channels/metabolism , Mitochondrial Proteins/metabolism , Adenosine Triphosphate/metabolism , Animals , Cardiomegaly/etiology , Constriction, Pathologic , Ion Channels/genetics , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Mitochondrial Proteins/genetics , Reactive Oxygen Species/metabolism , Uncoupling Protein 2
17.
Exp Ther Med ; 7(4): 947-952, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24669256

ABSTRACT

The aim of the present study was to investigate the effects of three blood purification methods on fibroblast growth factor-23 (FGF-23) clearance in patients with hyperphosphatemia undergoing maintenance hemodialysis (MHD). In addition, the correlation between serum FGF-23 and phosphorus (Pi) levels and the clinical implications were identified. Sixty-five MHD patients with hyperphosphatemia were randomly divided into three groups: Hemodialysis, HD (n=23); hemodiafiltration, HDF (n=21); and hemodialysis+hemoperfusion, HD+HP (n=21) groups. Serum Pi, FGF-23, blood urea nitrogen, serum creatinine and associated bio-marker levels were measured prior to and following treatment. The expression level of serum FGF-23 was observed to be positively correlated with Pi (r=0.45, P<0.01). The three blood purification methods that were adopted for the present study exhibited significant and effective clearance of serum Pi (P<0.05). The post-treatment serum FGF-23 levels were significantly decreased in the HDF and HD+HP groups (P<0.05). Therefore, HDF may be an effective method for clearing serum FGF-23 in MHD patients exhibiting hyperphosphatemia.

18.
Shanghai Kou Qiang Yi Xue ; 23(6): 651-3, 2014 Dec.
Article in Chinese | MEDLINE | ID: mdl-25636276

ABSTRACT

PURPOSE: To investigate the influences of repeated casting on the mechanical properties of CP Ti ceramic alloy. METHODS: CP Ti ceramic alloy samples were prepared and recast 3 times without adding any new CP Ti ceramic alloy. The physical properties of each specimen were measured. SPPPSS13.0 software package was used for statistical analysis. RESULTS: No significant difference was found on the flexural strength of the CP Ti ceramic alloys that had been cast 2 or 3 times, compared with that of the alloys being cast only 1 time (P>0.05). However, the flexural modulus, tensile strength, 0.2% yield strength and surface microhardness of the CP Ti alloys being cast 2 or 3 times were significantly higher than those of the alloys being cast only 1 time (P<0.05). Elongation of the CP Ti alloys being cast 2 or 3 times was significantly lower than that of the alloys being cast only 1 time (P<0.05). CONCLUSIONS: Recasting may cause decreases in tensile properties of CP Ti ceramic alloy.


Subject(s)
Ceramics , Dental Alloys , Hardness , Materials Testing , Tensile Strength , Titanium
19.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 29(3): 249-52, 2011 Jun.
Article in Chinese | MEDLINE | ID: mdl-21776847

ABSTRACT

OBJECTIVE: To study the mechanical properties of Co-Cr ceramic alloy after recasts. METHODS: Co-Cr ceramic alloy cast samples were prepared and recast for 3 times without adding any new Co-Cr ceramic alloy. The tensile strength, 0.2% yield strength, percentage of elongation, flexural strength, flexural modulus and Vickers hardness of each specimen were measured. RESULTS: Being cast for different times, the Co-Cr ceramic alloy showed no significant differences on their tensile strength, 0.2% yield strength, percentage of elongation, flexural strength, flexural modulus and Vickers hardness. CONCLUSION: Co-Cr ceramic alloy can be recast for 3 times at least, without decrease of the mechanical properties.


Subject(s)
Ceramics , Chromium Alloys , Alloys , Dental Stress Analysis , Hardness , Materials Testing , Metal Ceramic Alloys , Tensile Strength
20.
Zhonghua Kou Qiang Yi Xue Za Zhi ; 45(1): 36-8, 2010 Jan.
Article in Chinese | MEDLINE | ID: mdl-20368039

ABSTRACT

OBJECTIVE: To investigate the change of retentive forces of cast cobalt-chromium (Co-Cr) alloy clasp in cyclic fatigue test. METHODS: Samples of three types of cast Co-Cr alloy (Group A: Hardalloy; B: Regalloy™; C: Vera PDN™) clasps were fabricated and placed at undercut depths of 0.25 mm, 0.50 mm. The clasps were drawn from the model molar cyclicly to simulate 5 years of clinical use in an universal testing machine. Retentive force were record at 21 different time point for each clasp during the whole fatigue testing process. Data were subjected to ANOVA, Chi-square test and linear regression analysis. RESULTS: All clasps showed decreasing retention during the cyclic fatigue test. Clasps engaged in 0.50 mm undercut depth exhibited greater initial retentive force [Group A: (8.714 +/- 1.104) N, B: (9.072 +/- 0.653) N, C: (9.588 +/- 1.980) N] as well as greater loss of retention [Group A: (4.408 +/- 0.662) N, B: (3.484 +/- 0.494) N, C: (3.290 +/- 1.484) N] at the end of the test than clasps engaged in 0.25 mm undercut did [initial forces were (7.940 +/- 0.357), (7.834 +/- 1.308) and (8.156 +/- 1.067) N for Group A, B, C, respectively; loss of retention were (2.444 +/- 0.736) N, (2.954 +/- 1.048) N and (1.832 +/- 1.180) N for group A, B, C, respectively]. Negative correlation was found between the clasp retention and the logarithm of cycling times. CONCLUSIONS: Co-Cr alloy cast clasp could provide adequate retentive force for 5 years of clinical use.


Subject(s)
Chromium Alloys/chemistry , Dental Alloys/chemistry , Dental Clasps , Dental Casting Technique , Dental Stress Analysis , Denture Retention
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