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1.
Asian Journal of Andrology ; (6): 398-403, 2023.
Article in English | WPRIM | ID: wpr-981948

ABSTRACT

Teratozoospermia is a rare disease associated with male infertility. Several recurrent genetic mutations have been reported to be associated with abnormal sperm morphology, but the genetic basis of tapered-head sperm is not well understood. In this study, whole-exome sequencing (WES) identified a homozygous WD repeat domain 12 (WDR12; p.Ser162Ala/c.484T>G) variant in an infertile patient with tapered-head spermatozoa from a consanguineous Chinese family. Bioinformatic analysis predicted this mutation to be a pathogenic variant. To verify the effect of this variant, we analyzed WDR12 protein expression in spermatozoa of the patient and a control individual, as well as in the 293T cell line, by Western blot analysis, and found that WDR12 expression was significantly downregulated. To understand the role of normal WDR12, we evaluated its mRNA and protein expression in mice at different ages. We observed that WDR12 expression was increased in pachytene spermatocytes, with intense staining visible in round spermatid nuclei. Based on these results, the data suggest that the rare biallelic pathogenic missense variant (p.Ser162Ala/c.484T>G) in the WDR12 gene is associated with tapered-head spermatozoa. In addition, after intracytoplasmic sperm injection (ICSI), a successful pregnancy was achieved. This finding indicates that infertility associated with this WDR12 homozygous mutation can be overcome by ICSI. The present results may provide novel insights into understanding the molecular mechanisms of male infertility.


Subject(s)
Humans , Pregnancy , Female , Male , Animals , Mice , Teratozoospermia/pathology , Semen/metabolism , Infertility, Male/metabolism , Spermatozoa/metabolism , Mutation , RNA-Binding Proteins/metabolism , Cell Cycle Proteins/genetics
2.
Asian Journal of Andrology ; (6): 186-190, 2022.
Article in English | WPRIM | ID: wpr-928536

ABSTRACT

Nonobstructive azoospermia (NOA) is a common cause of infertility and is defined as the complete absence of sperm in ejaculation due to defective spermatogenesis. The aim of this study was to identify the genetic etiology of NOA in an infertile male from a Chinese consanguineous family. A homozygous missense variant of the membrane-bound O-acyltransferase domain-containing 1 (MBOAT1) gene (c.770C>T, p.Thr257Met) was found by whole-exome sequencing (WES). Bioinformatic analysis also showed that this variant was a pathogenic variant and that the amino acid residue in MBOAT1 was highly conserved in mammals. Quantitative polymerase chain reaction (Q-PCR) analysis showed that the mRNA level of MBOAT1 in the patient was 22.0% lower than that in his father. Furthermore, we screened variants of MBOAT1 in a broader population and found an additional homozygous variant of the MBOAT1 gene in 123 infertile men. Our data identified homozygous variants of the MBOAT1 gene associated with male infertility. This study will provide new insights for researchers to understand the molecular mechanisms of male infertility and will help clinicians make accurate diagnoses.


Subject(s)
Animals , Humans , Male , Acetyltransferases/genetics , Azoospermia/genetics , Cell Cycle Proteins/genetics , Infertility, Male/genetics , Mammals , Membrane Proteins/genetics , Mutation
3.
Journal of Central South University(Medical Sciences) ; (12): 1176-1184, 2020.
Article in English | WPRIM | ID: wpr-880583

ABSTRACT

OBJECTIVES@#To investigate the risk factors as well as their impact on patients' survival of central nervous system (CNS) complications following allogeneic hematopoietic stem cell transplantation (HSCT).@*METHODS@#All relevant clinical data from a total of 323 patients, who underwent allogeneic HSCT in Xiangya Hospital of Central South University from September 2016 to September 2019, were retrospectively reviewed in this study. The complications' occurrence time, common symptoms and some other clinical data of the patients who developed CNS complications were analyzed descriptively. The risk factors for CNS complications following allogeneic HSCT were analyzed through univariate and multivariate analysis. And the survival analysis was conducted as well.@*RESULTS@#Among the 323 patients who underwent allogeneic HSCT, 32 patients developed CNS complications. These complications occurred in these patients at a median of 32 (range from -1 to 584) d after transplantation. Common symptoms were disturbance of consciousness (78.1%), convulsion (59.4%), and headache (12.5%). Univariate analysis showed that there were significant differences in neutrophil engraftment, platelet (PLT) engraftment, serum cytomegalovirus (CMV) DNA positive, combined with acute graft-versus-host disease (aGVHD), donor selection (@*CONCLUSIONS@#The delay or the failure of PLT engraftment and combined with aGVHD are the risk factors for CNS complications. The facts indicate that we should prevent CNS complications when patients who underwent allogeneic HSCT with the delay or the failure of PLT engraftment or aGVHD. Compared with non-CNS complication group, patients who developed CNS complications usually have poor prognosis.


Subject(s)
Humans , Central Nervous System , Graft vs Host Disease/etiology , Hematopoietic Stem Cell Transplantation/adverse effects , Retrospective Studies , Risk Factors
4.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 892-899, 2019.
Article in English | WPRIM | ID: wpr-781241

ABSTRACT

Obesity that is highly associated with numerous metabolic diseases has become a global health issue nowdays. Plant sesterterpenoids are an important group of natural products with great potential; thus, their bioactivities deserve extensive exploration. RNA-seq analysis indicated that leucosceptroid B, a sesterterpenoid previously discovered from the glandular trichomes of Leucosceptrum canum, significantly regulated the expression of 10 genes involved in lipid metabolism in Caenorhabditis elegans. Furthermore, leucosceptroid B was found to reduce fat storage, and downregulate the expression of two stearoyl-CoA desaturase (SCD) genes fat-6 and fat-7, and a fatty acid elongase gene elo-2 in wild-type C. elegans. In addition, leucosceptroid B significantly decreased fat accumulation in both fat-6 and fat-7 mutant worms but did not affect the fat storage of fat-6; fat-7 double mutant. These findings indicated that leucosceptroid B reduced fat storage depending on the downregulated expression of fat-6, fat-7 and elo-2 and thereby inhibiting the biosynthesis of the corresponding unsaturated fatty acid. These findings provide new insights into the development and utilization of plant sesterterpenoids as potential antilipemic agents.

5.
The Korean Journal of Parasitology ; : 325-334, 2018.
Article in English | WPRIM | ID: wpr-742267

ABSTRACT

Toxoplasma gondii is an apicomplexan zoonotic protozoan parasite that infects most species of warm-blooded animals, including humans. The heavy incidence and severe or lethal damage caused by T. gondii infection clearly indicate a need for the development of an effective vaccine. T. gondii GRA8 is a member of the dense granules protein family and is used as a marker of acute infection. In the present study, we evaluated the protective immunity induced by DNA vaccination based on a recombinant eukaryotic plasmid, pDsRed2-GRA8, against acute toxoplasmosis in mice. BALB/c mice were intramuscularly immunized with the pDsRed2-GRA8 plasmid and then challenged by infection with the highly virulent GFP-RH strain of T. gondii. The specific immune responses and protective efficacy against T. gondii of this vaccine were analyzed by measuring cytokine and serum antibody titers, splenocyte proliferation assays, and the survival times of mice after challenge. Our results showed that mice immunized with pDsRed2-GRA8 demonstrated specific humoral and cellular responses, induced higher IgG antibody titers with predominant IgG2a production; increased levels of IL-10, IL-12 (p70), IFN-γ, TNF-α, and splenocyte proliferation; and prolonged survival times compared to those of control mice. The present study showed that DNA immunization with pDsRed2-GRA8 induced humoral and cellular immune responses, and all immunized mice showed greater Th1-type immune responses and longer survival times than those of control mice. These results indicated that T. gondii GRA8 DNA immunization induces a partial protective effect against acute toxoplasmosis.


Subject(s)
Animals , Humans , Mice , DNA , Immunity, Cellular , Immunization , Immunoglobulin G , Incidence , Interleukin-10 , Interleukin-12 , Parasites , Plasmids , Toxoplasma , Toxoplasmosis , Vaccination
6.
The Korean Journal of Parasitology ; : 135-145, 2018.
Article in English | WPRIM | ID: wpr-742239

ABSTRACT

Due to the critical location and physiological activities of the retinal pigment epithelial (RPE) cell, it is constantly subjected to contact with various infectious agents and inflammatory mediators. However, little is known about the signaling events in RPE involved in Toxoplasma gondii infection and development. The aim of the study is to screen the host mRNA transcriptional change of 3 inflammation-related gene categories, PI3K/Akt pathway regulatory components, blood vessel development factors and ROS regulators, to prove that PI3K/Akt or mTOR signaling pathway play an essential role in regulating the selected inflammation-related genes. The selected genes include PH domain and leucine- rich-repeat protein phosphatases (PHLPP), casein kinase2 (CK2), vascular endothelial growth factor (VEGF), pigment epithelium-derived factor (PEDF), glutamate-cysteine ligase (GCL), glutathione S-transferase (GST), and NAD(P)H: quinone oxidoreductase (NQO1). Using reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), we found that T. gondii up-regulates PHLPP2, CK2β, VEGF, GCL, GST, and NQO1 gene expression levels, but down-regulates PHLPP1 and PEDF mRNA transcription levels. PI3K inhibition and mTOR inhibition by specific inhibitors showed that most of these host gene expression patterns were due to activation of PI3K/Akt or mTOR pathways with some exceptional cases. Taken together, our results reveal a new molecular mechanism of these gene expression change dependent on PI3K/Akt or mTOR pathways and highlight more systematical insight of how an intracellular T. gondii can manipulate host genes to avoid host defense.


Subject(s)
Blood Vessels , Caseins , Epithelial Cells , Gene Expression , Glutamate-Cysteine Ligase , Glutathione Transferase , Hydrogen-Ion Concentration , Phosphoprotein Phosphatases , Polymerase Chain Reaction , Reactive Oxygen Species , Retinaldehyde , Reverse Transcription , RNA, Messenger , Signal Transduction , Toxoplasma , Toxoplasmosis , Vascular Endothelial Growth Factor A
7.
The Korean Journal of Parasitology ; : 95-98, 2017.
Article in English | WPRIM | ID: wpr-168697

ABSTRACT

Fasciola hepatica is a trematode that causes zoonosis, mainly in cattle and sheep, and occasionally in humans. Few recent studies have determined the infection status of this fluke in Korea. In August 2015, we collected 402 samples of freshwater snails at Hoenggye-ri (upper stream) and Suha-ri (lower stream) of Song-cheon (stream) in Daegwalnyeong-myeon, Pyeongchang-gun in Gangwon-do (Province) near many large cattle or sheep farms. F. hepatica infection was determined using PCR on the nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 402 samples, F. hepatica 1TS-2 marker was detected in 6 freshwater snails; thus, the overall prevalence in freshwater snails was 1.5%. The prevalence varied between collection areas, ranging from 0.0% at Hoenggye-ri to 2.9% at Suha-ri. However, F. gigantica ITS-2 was not detected in the 6 F. hepatica-positive samples by PCR. The nucleotide sequences of the 6 F. hepatica ITS-2 PCR-positive samples were 99.4% identical to the F. hepatica ITS-2 sequences in GenBank, whereas they were 98.4% similar to F. gigantica ITS-2 sequences. These results indicated that the prevalence of F. hepatica in snail intermediate hosts was 1.5% in Gangwon-do, Korea; however the prevalence varied between collection areas. These results may help us to understand F. hepatica infection status in natural environments.


Subject(s)
Animals , Cattle , Humans , Agriculture , Base Sequence , Databases, Nucleic Acid , Fasciola hepatica , Fasciola , Fresh Water , Korea , Polymerase Chain Reaction , Prevalence , Ranunculaceae , Sheep , Snails , Trematoda
8.
Chinese Medical Journal ; (24): 979-985, 2017.
Article in English | WPRIM | ID: wpr-266877

ABSTRACT

<p><b>BACKGROUND</b>Several studies concerning the association between glutathione S-transferase P1 (GSTP1) Ile105Val polymorphism and male infertility risk have reported controversial findings. The present study was aimed to explore this association using a meta-analysis.</p><p><b>METHODS</b>The PubMed, EMBASE, China National Knowledge Infrastructure (CNKI), and Wanfang databases were searched. Odds ratios (OR s) with 95% confidence intervals (CI s) were calculated to estimate the strength of the association.</p><p><b>RESULTS</b>A total of 3282 cases and 3268 controls in nine case-control studies were included. There was no significant association between GSTP1 Ile105Val polymorphism and male infertility in the overall population, but significant associations were found under the dominant (OR = 1.23, 95% CI = 1.04-1.46, I2 = 32.2%) and heterozygote (OR = 1.29, 95% CI = 1.08-1.53, I2 = 26.8%) models after excluding studies for which the data did not satisfy Hardy-Weinberg equilibrium (HWE). Similarly, subgroup analyses revealed no significant association in Asians or Chinese population although a significant association was apparent among Chinese population in studies with HWE under the heterozygote model (OR = 1.25, 95% CI = 1.03-1.52, I2 = 44.1%). Significant heterogeneity could be observed in some genetic models, but this heterogeneity was not significant when stratified by HWE. No evidence for publication bias was found.</p><p><b>CONCLUSIONS</b>The GSTP1 Ile105Val polymorphism might not be associated with male infertility risk, and thus additional well-designed studies with larger sample size are warranted.</p>


Subject(s)
Humans , Male , Asian People , Genetic Association Studies , Genetic Predisposition to Disease , Glutathione S-Transferase pi , Genetics , Infertility, Male , Genetics , Polymorphism, Single Nucleotide , Genetics
9.
An. bras. dermatol ; 91(5,supl.1): 76-78, Sept.-Oct. 2016. tab, graf
Article in English | LILACS | ID: biblio-837916

ABSTRACT

Abstract Cutaneous reactions associated with interferons (IFNs) treatment are either localized or generalized. The most common presentation of localized reactions at IFNs injection site is usually an erythematous patch or plaque. Local leukocytoclastic vasculitis presenting with cutaneous necrosis is extremely rare. We report a 19-year-old man with hepatitis B who had local leukocytoclastic vasculitis induced by interferon-gama injection at the injection site. After changing the injection sites and using the combined treatment of prednisone and colchicine, the previous lesion healed and no other cutaneous lesion occurred. We also made a mini review of such cases.


Subject(s)
Humans , Male , Young Adult , Skin/pathology , Interferon-gamma/adverse effects , Vasculitis, Leukocytoclastic, Cutaneous/chemically induced , Skin/drug effects , Prednisone/therapeutic use , Colchicine/therapeutic use , Treatment Outcome , Vasculitis, Leukocytoclastic, Cutaneous/pathology , Vasculitis, Leukocytoclastic, Cutaneous/drug therapy , Erythema/chemically induced , Erythema/pathology , Injections, Subcutaneous/adverse effects , Anti-Inflammatory Agents/therapeutic use , Necrosis/chemically induced , Necrosis/pathology
10.
Journal of Clinical Hepatology ; (12): 992-996, 2016.
Article in Chinese | WPRIM | ID: wpr-778645

ABSTRACT

When hepatitis B virus (HBV) invades the human body, innate immunity acts the earliest and plays an important role. When an adult is infected with HBV, HBV can often be eliminated spontaneously. However, if HBV infection occurs right after birth or when the patient is young, the disease tends to become chronic and affect the whole life. In the incipient stage of HBV infection, innate immune response plays an important role in inhibiting viral replication, and the prognosis of HBV infection depends on the combined effect of host and virus. This article briefly introduces the research advances in the cells, cytokines, and signaling pathways that play important roles in the host's innate immunity against HBV and their mechanisms of action, and points out their potential values in clinical treatment.

11.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 372-376, 2016.
Article in English | WPRIM | ID: wpr-285260

ABSTRACT

Infection of schistosomiasis japonica may eventually lead to liver fibrosis, and no effective antifibrotic therapies are available but liver transplantation. Hedgehog (HH) signaling pathway has been involved in the process and is a promising target for treating liver fibrosis. This study aimed to explore the effects of pentoxifylline (PTX) on liver fibrosis induced by schistosoma japonicum infection by inhibiting the HH signaling pathway. Phorbol12-myristate13-acetate (PMA) was used to induce human acute mononuclear leukemia cells THP-1 to differentiate into macrophages. The THP-1-derived macrophages were stimulated by soluble egg antigen (SEA), and the culture supernatants were collected for detection of activation of macrophages. Cell Counting Kit-8 (CCK-8) was used to detect the cytotoxicity of the culture supernatant and PTX on the LX-2 cells. The LX-2 cells were administered with activated culture supernatant from macrophages and(or) PTX to detect the transforming growth factor-β gene expression. The mRNA expression of shh and gli-1, key parts in HH signaling pathway, was detected. The mRNA expression of shh and gli-1 was increased in LX-2 cells treated with activated macrophages-derived culture supernatant, suggesting HH signaling pathway may play a key role in the activation process of hepatic stellate cells (HSCs). The expression of these genes decreased in LX-2 cells co-cultured with both activated macrophages-derived culture supernatant and PTX, indicating PTX could suppress the activation process of HSCs. In conclusion, these data provide evidence that PTX prevents liver fibrogenesis in vitro by the suppression of HH signaling pathway.


Subject(s)
Animals , Humans , Antigens, Helminth , Pharmacology , Cell Culture Techniques , Cell Differentiation , Cell Line , Culture Media, Conditioned , Chemistry , Pharmacology , Gene Expression Regulation , Hedgehog Proteins , Genetics , Allergy and Immunology , Hepatic Stellate Cells , Cell Biology , Metabolism , Liver Cirrhosis , Metabolism , Parasitology , Macrophage Activation , Macrophages , Cell Biology , Allergy and Immunology , Models, Biological , Monocytes , Cell Biology , Metabolism , Pentoxifylline , Pharmacology , Phosphodiesterase Inhibitors , Pharmacology , RNA, Messenger , Genetics , Allergy and Immunology , Schistosoma japonicum , Chemistry , Signal Transduction , Tetradecanoylphorbol Acetate , Pharmacology , Zinc Finger Protein GLI1 , Genetics , Allergy and Immunology , Zygote , Chemistry
12.
The Korean Journal of Parasitology ; : 711-717, 2016.
Article in English | WPRIM | ID: wpr-72763

ABSTRACT

Toxoplasma gondii is an obligate intracellular parasite that stimulates production of high levels of proinflammatory cytokines, which are important for innate immunity. NLRs, i.e., nucleotide-binding oligomerization domain (NOD)-like receptors, play a crucial role as innate immune sensors and form multiprotein complexes called inflammasomes, which mediate caspase-1-dependent processing of pro-IL-1β. To elucidate the role of inflammasome components in T. gondii-infected THP-1 macrophages, we examined inflammasome-related gene expression and mechanisms of inflammasome-regulated cytokine IL-1β secretion. The results revealed a significant upregulation of IL-1β after T. gondii infection. T. gondii infection also upregulated the expression of inflammasome sensors, including NLRP1, NLRP3, NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and NAIP, in a time-dependent manner. The infection also upregulated inflammasome adaptor protein ASC and caspase-1 mRNA levels. From this study, we newly found that T. gondii infection regulates NLRC4, NLRP6, NLRP8, NLRP13, AIM2, and neuronal apoptosis inhibitor protein (NAIP) gene expressions in THP-1 macrophages and that the role of the inflammasome-related genes may be critical for mediating the innate immune responses to T. gondii infection.


Subject(s)
Apoptosis , Cytokines , Gene Expression , Immunity, Innate , Inflammasomes , Macrophages , Multiprotein Complexes , Negotiating , Neurons , Parasites , RNA, Messenger , Toxoplasma , Up-Regulation
13.
Journal of Forensic Medicine ; (6): 424-427, 2016.
Article in Chinese | WPRIM | ID: wpr-984871

ABSTRACT

OBJECTIVES@#To study the changes of alcohol content and pharmacokinetic parameter in rats after taking Huoxiang Zhengqi liquid.@*METHODS@#The rats were randomly divided into three groups and given with white alcohol at the dose of 3.0 mL/kg, low-dose and high-dose Chinese medicine liquor, respectively. The blood was collected before administration and 5 min, 10 min, 15 min, 30 min, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h and 8 h after administration by cutting rats' tails. The concentrations of alcohol in blood were detected by headspace-gas chromatography method. The main pharmacokinetic parameters were calculated by DAS 2.0, and then analyzed by SPSS 17.0.@*RESULTS@#The difference of maximum blood concentrations between high-dose Chinese medicine alcohol group and white alcohol group was statistically significant (P<0.05). There was no significant difference in other pharmacokinetic parameters among three groups (P>0.05).@*CONCLUSIONS@#The Chinese herbal medicinal ingredients in the Huoxiang Zhengqi liquid has no effect on the metabolism and elimination of ethanol in rats. The research provides useful reference for the qualitative assessment and processing of traffic accident cases involved in Huoxiang Zhengqi liquid and the studies related to drug-interaction.


Subject(s)
Animals , Rats , Drugs, Chinese Herbal/pharmacology , Ethanol/pharmacokinetics
14.
International Eye Science ; (12): 681-683, 2015.
Article in Chinese | WPRIM | ID: wpr-637244

ABSTRACT

AIM: To investigate the preoperative binocular visual function of intermittent exotropia and the rebuilding and recovery of the postoperative binocular visual function, and analyze the effect of binocular visual function on orthophoria after surgery. METHODS:From January 2011 to January 2014, 47 basic intermittent exotropia patients caming for treatment were collected in the clinical data. The changes in their near stereopsis, binocular visual function, binocular fusion and distance stereopsis after operations were recorded in the form of data. The preoperative binocular vision and the postoperative rebuilding were analyzed and contrasted with each other. In addition, the effect on the postoperative maintaining of orthophoria due to the existence, recovery and rebuilding of binocular visual function were observed. RESULTS:Intermittent exotropia patients got damage in different levels on their binocular visual functions, especially on distance stereopsis, which was the heaviest and earliest. After the operation, all functions were obviously recovered and reconstructed and the improvements were statistically significant compared against those before the operation (PCONCLUSION: The intermittent exotropia cause damage to the stereopsis which happened the earliest. Obvious recovery and reconstruction of binocular visual function can be observed after the surgery. A relatively good preoperative binocular visual function may lead to the increase in the ratio of orthophoria or cure the intermittent exotropia. Performing an operation when distance stereopsis is damaged can increase the success rate for the surgery and reduce the recurrence rate.

15.
The Korean Journal of Parasitology ; : 641-645, 2015.
Article in English | WPRIM | ID: wpr-180023

ABSTRACT

Fascioliasis, a food-borne trematode zoonosis, is a disease primarily in cattle and sheep and occasionally in humans. Water dropwort (Oenanthe javanica), an aquatic perennial herb, is a common second intermediate host of Fasciola, and the fresh stems and leaves are widely used as a seasoning in the Korean diet. However, no information regarding Fasciola species contamination in water dropwort is available. Here, we collected 500 samples of water dropwort in 3 areas in Korea during February and March 2015, and the water dropwort contamination of Fasciola species was monitored by DNA sequencing analysis of the Fasciola hepatica and Fasciola gigantica specific mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 500 samples assessed, the presence of F. hepatica cox1 and 1TS-2 markers were detected in 2 samples, and F. hepatica contamination was confirmed by sequencing analysis. The nucleotide sequences of cox1 PCR products from the 2 F. hepatica-contaminated samples were 96.5% identical to the F. hepatica cox1 sequences in GenBank, whereas F. gigantica cox1 sequences were 46.8% similar with the sequence detected from the cox1 positive samples. However, F. gigantica cox1 and ITS-2 markers were not detected by PCR in the 500 samples of water dropwort. Collectively, in this survey of the water dropwort contamination with Fasciola species, very low prevalence of F. hepatica contamination was detected in the samples.


Subject(s)
Animals , Base Sequence , Cluster Analysis , DNA, Helminth/chemistry , DNA, Ribosomal Spacer/chemistry , Electron Transport Complex IV/genetics , Fasciola hepatica/genetics , Korea , Molecular Sequence Data , Oenanthe/parasitology , Phylogeny , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid
16.
The Korean Journal of Parasitology ; : 271-277, 2015.
Article in English | WPRIM | ID: wpr-83622

ABSTRACT

The genetic diversity of Schistosoma haematobium remains largely unstudied in comparison to that of Schistosoma mansoni. To characterize the extent of genetic diversity in S. haematobium among its definitive host (humans), we collected S. haematobium eggs from the urine of 73 infected schoolchildren at 5 primary schools in White Nile State, Sudan, and then performed a randomly amplified polymorphic DNA marker ITS2 by PCR-RFLP analysis. Among 73 S. haematobium egg-positive cases, 13 were selected based on the presence of the S. haematobium satellite markers A4 and B2 in their genomic DNA, and used for RFLP analysis. The 13 samples were subjected to an RFLP analysis of the S. haematobium ITS2 region; however, there was no variation in size among the fragments. Compared to the ITS2 sequences obtained for S. haematobium from Kenya, the nucleotide sequences of the ITS2 regions of S. haematobium from 4 areas in Sudan were consistent with those from Kenya (> 99%). In this study, we demonstrate for the first time that most of the S. haematobium population in Sudan consists of a pan-African S. haematobium genotype; however, we also report the discovery of Kenyan strain inflow into White Nile, Sudan.


Subject(s)
Adolescent , Animals , Child , Female , Humans , Male , Base Sequence , DNA, Helminth/genetics , Genetic Variation , Genotype , Molecular Sequence Data , Ovum/classification , Parasite Egg Count , Polymorphism, Restriction Fragment Length , Schistosoma haematobium/genetics , Schistosomiasis haematobia/diagnosis , Students , Sudan/epidemiology , Urine/parasitology
17.
The Korean Journal of Parasitology ; : 371-377, 2015.
Article in English | WPRIM | ID: wpr-50468

ABSTRACT

Trichomonas vaginalis induces proinflammation in cervicovaginal mucosal epithelium. To investigate the signaling pathways in TNF-alpha production in cervical mucosal epithelium after T. vaginalis infection, the phosphorylation of PI3K/AKT and MAPK pathways were evaluated in T. vaginalis-infected SiHa cells in the presence and absence of specific inhibitors. T. vaginalis increased TNF-alpha production in SiHa cells, in a parasite burden-dependent and incubation time-dependent manner. In T. vaginalis-infected SiHa cells, AKT, ERK1/2, p38 MAPK, and JNK were phosphorylated from 1 hr after infection; however, the phosphorylation patterns were different from each other. After pretreatment with inhibitors of the PI3K/AKT and MAPK pathways, TNF-alpha production was significantly decreased compared to the control; however, TNF-alpha reduction patterns were different depending on the type of PI3K/MAPK inhibitors. TNF-alpha production was reduced in a dose-dependent manner by treatment with wortmannin and PD98059, whereas it was increased by SP600125. These data suggested that PI3K/AKT and MAPK signaling pathways are important in regulation of TNF-alpha production in cervical mucosal epithelial SiHa cells. However, activation patterns of each pathway were different from the types of PI3K/MAPK pathways.


Subject(s)
Female , Humans , Cell Line , Cervix Uteri/enzymology , Epithelial Cells/enzymology , MAP Kinase Signaling System , Mucous Membrane/enzymology , Phosphatidylinositol 3-Kinases/genetics , Proto-Oncogene Proteins c-akt/genetics , Trichomonas Vaginitis/enzymology , Trichomonas vaginalis/physiology , Tumor Necrosis Factor-alpha/genetics
18.
The Korean Journal of Parasitology ; : 645-652, 2014.
Article in English | WPRIM | ID: wpr-124060

ABSTRACT

Fasciola hepatica is a trematode that causes zoonosis mainly in cattle and sheep and occasionally in humans. Fascioliasis has been reported in Korea; however, determining F. hepatica infection in snails has not been done recently. Thus, using PCR, we evaluated the prevalence of F. hepatica infection in snails at 4 large water-dropwort fields. Among 349 examined snails, F. hepatica-specific internal transcribed space 1 (ITS-1) and/or ITS-2 markers were detected in 12 snails and confirmed using sequence analysis. Morphologically, 213 of 349 collected snails were dextral shelled, which is the same aperture as the lymnaeid snail, the vectorial host for F. hepatica. Among the 12 F. hepatica-infected snails, 6 were known first intermediate hosts in Korea (Lymnaea viridis and L. ollula) and the remaining 6 (Lymnaea sp.) were potentially a new first intermediate host in Korea. It has been shown that the overall prevalence of the snails contaminated with F. hepatica in water-dropwort fields was 3.4%; however, the prevalence varied among the fields. This is the first study to estimate the prevalence of F. hepatica infection using the vectorial capacity of the snails in Korea.


Subject(s)
Animals , Base Sequence , DNA, Helminth/chemistry , DNA, Ribosomal Spacer/chemistry , Fasciola hepatica/anatomy & histology , Molecular Sequence Data , Oenanthe/growth & development , Polymerase Chain Reaction , Republic of Korea , Sequence Analysis, DNA , Snails/growth & development
19.
The Korean Journal of Parasitology ; : 595-603, 2014.
Article in English | WPRIM | ID: wpr-229078

ABSTRACT

Trichomonas vaginalis secretes a number of proteases which are suspected to be the cause of pathogenesis; however, little is understood how they manipulate host cells. The mammalian target of rapamycin (mTOR) regulates cell growth, cell proliferation, cell motility, cell survival, protein synthesis, and transcription. We detected various types of metalloproteinases including GP63 protein from T. vaginalis trophozoites, and T. vaginalis GP63 metalloproteinase was confirmed by sequencing and western blot. When SiHa cells were stimulated with live T. vaginalis, T. vaginalis excretory-secretory products (ESP) or T. vaginalis lysate, live T. vaginalis and T. vaginalis ESP induced the mTOR cleavage in both time- and parasite load-dependent manner, but T. vaginalis lysate did not. Pretreatment of T. vaginalis with a metalloproteinase inhibitor, 1,10-phenanthroline, completely disappeared the mTOR cleavage in SiHa cells. Collectively, T. vaginalis metallopeptidase induces host cell mTOR cleavage, which may be related to survival of the parasite.


Subject(s)
Humans , Blotting, Western , Cell Line, Tumor , Epithelial Cells/metabolism , Metalloproteases/genetics , Proteolysis , Sequence Analysis, DNA , TOR Serine-Threonine Kinases/metabolism , Trichomonas vaginalis/enzymology
20.
China Journal of Chinese Materia Medica ; (24): 3902-3906, 2013.
Article in Chinese | WPRIM | ID: wpr-319682

ABSTRACT

To optimize the separation process of liquirtin from glycyrrhiz by static, dynamic adsorption and desorption experiments on polyamide resin, with liquirtin, isoliquiritin and glycyrrhizic acid as the study index. The optimum process conditions were that the pH of solution was regulated to be 7.0, the concentration of liquirtin was 1.296 g x L(-1), the volume of loading buffer was 3 BV. After absorption, efforts shall be made to elute resin with water, 10%, 20%, 30% ethanol (3 BV for each), collect 20% ethanol eluted fraction, and recover solvents. The results showed lower contents of such impurities as isoliquiritin and isoliquiritin in extracts sepaprated under this process conditions, as well as an increase in purity of liquirtin from 4.86% to 88.5%. The method was simple and feasible, it could obtain a higher purity in extracts from liquirtin and provide basis for industrialized separation and preparation of liquirtin.


Subject(s)
Chalcone , Drugs, Chinese Herbal , Glucosides , Glycyrrhiza uralensis , Chemistry , Glycyrrhizic Acid , Resins, Synthetic , Chemistry
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