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1.
National Journal of Andrology ; (12): 980-984, 2009.
Artículo en Chino | WPRIM | ID: wpr-252846

RESUMEN

<p><b>OBJECTIVE</b>To investigate the difference in the responsiveness of intracellular free Ca2+ concentration ([Ca2+]i) to progesterone in the spermatozoa of normal fertile men and patients with unexplained infertility.</p><p><b>METHODS</b>Nine normal fertile men and 10 patients with unexplained infertility were selected in this study. After swim-up separation of the motile fraction and 2-hour in vitro capacitation, the spermatozoa were loaded with the fluorescent calcium indicator Fluo-3/AM (8.85 micromol/L) for 40 minutes away from the light, and then the sperm suspension was mixed with equal amount of 20% gelatin to immobilize the spermatozoa. The basal intracellular free [Ca2+]i and that induced by 10 micromol/L progesterone in the individual sperm were assessed by laser scanning confocal microscopy.</p><p><b>RESULTS</b>The infertile patients had a significantly lower basal level of [Ca2+]i in the capacitated sperm than the fertile men (P < 0.01). The sperm from the normal controls responded to progesterone by exhibiting a rapid but transient rise in [Ca2+]i, with the peak level significantly higher than the basal level (P < 0.05), while those from the infertile patients by showing a slight increase, with no significant difference between the peak and basal levels (P > 0.05). Both the peak of the progesterone-induced [Ca2+]i and its increase amplitude expressed as the difference between the peak and basal levels were significantly higher in the normal fertile group than in the infertile patients (P < 0.01).</p><p><b>CONCLUSION</b>The responsiveness of [Ca2+]i to progesterone is reduced in the spermatozoa of patients with unexplained infertility, which suggests a functional defect in the non-genomic sperm membrane progesterone receptor responsible for calcium influx.</p>


Asunto(s)
Adulto , Femenino , Humanos , Masculino , Adulto Joven , Reacción Acrosómica , Calcio , Estudios de Casos y Controles , Infertilidad Masculina , Progesterona , Farmacología , Espermatozoides
2.
Chinese Journal of Pathology ; (12): 292-295, 2006.
Artículo en Chino | WPRIM | ID: wpr-277417

RESUMEN

<p><b>OBJECTIVE</b>To screen for siRNAs that inhibit the expression of p42(MAPK) in HeLa cell line.</p><p><b>METHODS</b>Three p42(MAPK) siRNAs and one random siRNA were synthesized using Silencer siRNA Construction Kit, and labeled with Cy-3 for measurement of transfection effect. SiRNAs were transfected into HeLa cells by Lipofectamin 2000. The expression of p42(MAPK) was analyzed by Western blot. The biological effect of siRNAs on HeLa cell growth was monitored by MTT and flow cytometry.</p><p><b>RESULTS</b>Two siRNAs (siRNA-2 and siRNA-3) among three tested were identified to be able to downregulate the p42(MAPK) expression. A concurrent growth retardation of HeLa cell line was observed in comparison with the control.</p><p><b>CONCLUSION</b>Inhibition of p42(MAPK) expression with siRNA technique can inhibit the proliferation of HeLa cells.</p>


Asunto(s)
Humanos , Ciclo Celular , Proliferación Celular , Supervivencia Celular , Citometría de Flujo , Células HeLa , Proteína Quinasa 1 Activada por Mitógenos , Genética , Interferencia de ARN , ARN Interferente Pequeño , Genética , Transfección
3.
Chinese Journal of Medical Genetics ; (6): 627-630, 2006.
Artículo en Chino | WPRIM | ID: wpr-285064

RESUMEN

<p><b>OBJECTIVE</b>To investigate the protocol of the construction of HERG gene mutations, an A561V mutation which was detected in a Chinese congenital long QT syndrome (LQTS) family had been constructed and expressed in vitro.</p><p><b>METHODS</b>The A561V cloning vector PGEM-HERG-A561V was constructed by quick site-directed mutagenesis PCR. The A561V expressive vector pcDNA3-HERG-A561V was constructed by restriction enzymes. pRK5-GFP was cotransfected with pcDNA3-HERG-A561V or wild type pcDNA3-HERG into HEK293 cells by Superfect transfection reagent. The protein was measured by immunofluorescence.</p><p><b>RESULTS</b>Direct sequence analyses revealed a C to T transition at position 1682. The A561V mutation was correctly combined to eukaryotic expressive vector pcDNA3 and expressed in HEK293 cells. The protein of mutation was expressed in cytoplasm and cellular membrane while the wild type gene was expressed only on cellular membrane.</p><p><b>CONCLUSION</b>The protocol can be used successfully to construct and express HERG A561V mutation and it forms the basement of the further study on functions of mutation.</p>


Asunto(s)
Humanos , Secuencia de Bases , Línea Celular , Membrana Celular , Metabolismo , Citoplasma , Metabolismo , ADN , Química , Genética , Análisis Mutacional de ADN , Canal de Potasio ERG1 , Canales de Potasio Éter-A-Go-Go , Genética , Metabolismo , Vectores Genéticos , Genética , Proteínas Fluorescentes Verdes , Genética , Metabolismo , Síndrome de QT Prolongado , Genética , Microscopía Fluorescente , Mutación Puntual , Proteínas Recombinantes de Fusión , Genética , Metabolismo , Transfección
4.
China Journal of Chinese Materia Medica ; (24): 357-360, 2005.
Artículo en Chino | WPRIM | ID: wpr-279163

RESUMEN

<p><b>OBJECTIVE</b>To study the effect of 20( R)-ginsenoside Rg3 on the expressions of angiogenesis factors proteins (VEGF,bFGF, MMP-2) in human lung adenocarcinoma cell line A549 and HUVEC304 cell.</p><p><b>METHOD</b>The cell lines of A549 and HUVEC304 were cultured with 20(R)- Rg3. The gray scale and positive rate of VEGF, bFGF, MMP-2 were detected by immunohistochemistry. The differential expressions of genes were studied by DNA microarray.</p><p><b>RESULT</b>The positive rate of VEGF protein in A549 cell decreased significantly as compared with the control group ( P = 0.03). The gray scales of VEGF, Flt, KDT proteins in both A549 cell lines and HUVEC 304 cell lines decreased ( P = 0.05). Gray scale of MMP-2 also decreased in A549 cell lines. The result of differential expressions of genes of A549 cell lines showed that 14 genes were down-regulated and 10 genes were up-regulated.</p><p><b>CONCLUSION</b>The Chinese materia medica of 20( R)-Rg3 can inhibit the expression of angiogenesis factors proteins via several target genes in both tumour cell and vascular endothelial cell.</p>


Asunto(s)
Humanos , Adenocarcinoma , Genética , Metabolismo , Patología , Línea Celular Tumoral , Células Endoteliales , Metabolismo , Perfilación de la Expresión Génica , Ginsenósidos , Farmacología , Neoplasias Pulmonares , Genética , Metabolismo , Patología , Metaloproteinasa 2 de la Matriz , Metabolismo , Neovascularización Patológica , Análisis de Secuencia por Matrices de Oligonucleótidos , Panax , Química , Venas Umbilicales , Biología Celular , Factor A de Crecimiento Endotelial Vascular , Metabolismo
5.
Chinese Journal of Plastic Surgery ; (6): 48-50, 2004.
Artículo en Chino | WPRIM | ID: wpr-327326

RESUMEN

<p><b>OBJECTIVE</b>To observe the effects of NGF, estrogens and minoxidil on the growth of human hair follicle in vitro.</p><p><b>METHODS</b>In a model of human hair follicle in vitro, the follicle was separately treated with the NGF, estrogens and minoxidil. The growth of the hair follicle was measured in length with an eyepiece micrometer. The effects of the NGF, estrogens and minoxidil were evaluated by measuring the rates of incorporation of 3H-TdR of DNA synthesis.</p><p><b>RESULTS</b>The growth of the human hair follicle was showing significantly faster in the 100 ng/ml NGF and 125 micrograms/ml minoxidil groups, compared with the control (P < 0.05), but the growth was significantly inhibited in the 0.5 microgram/ml 17 beta-E2 group (P < 0.05). There was no difference shown for the growth of the hair follicle in the group mixed with 100 ng/ml NGF and 0.5 microgram/ml 17 beta-E2 (P > 0.05). The rates of incorporation of 3H-TdR in the groups were shown that the results just correlated with the results of the above-mentioned method.</p><p><b>CONCLUSIONS</b>The 100 ng/ml NGF and 125 micrograms/ml minoxidil could increase the growth of human hair follicle while the 0.5 microgram/ml 17 beta-E2 could inhibit it. The 100 ng/ml NGF could neutralized the effect of the 0.5 microgram/ml 17 beta-E2.</p>


Asunto(s)
Adolescente , Adulto , Humanos , Persona de Mediana Edad , Estrógenos , Farmacología , Folículo Piloso , Biología Celular , Técnicas In Vitro , Minoxidil , Farmacología , Factor de Crecimiento Nervioso , Farmacología , Vasodilatadores , Farmacología
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