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1.
Chinese journal of integrative medicine ; (12): 51-58, 2019.
Artículo en Inglés | WPRIM | ID: wpr-773987

RESUMEN

OBJECTIVE@#To investigate the potential antifibrotic mechanisms of Chinese medicine Ganshuang Granules (, GSG) and to provide clinical therapeutic evidence of its effects.@*METHODS@#A cirrhotic mouse model was established by intraperitoneally injecting a mixture of CCl (40%) and oil (60%) at 0.2 mL per 100 g of body weight twice a week for 12 weeks. After 12-week modeling, GSG was intragastric administrated to the mice for 2 weeks, and the mice were divided into low-, medium- and high-dose groups at doses of 1, 2 and 4 g/(kg·day), respectively. Liver morphology changes were observed using Masson's trichrome staining and B-ultrasound. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and hyaluronic acid (HA) in serum were detected using an automatic biochemistry analyzer. The expressions of desmin, smooth muscle actin (SMA) and Foxp3 in liver were detected by immunoflfluorescence. The regulatory T cell (Treg) frequency was determined through flflow cytometry analysis. Collagen-I, SMA, IL-6, tumor necrosis factor α (TNF-α), interleukin (IL)-1β and transforming growth factor β1 (TGF-β1) expression levels were measured using quantitative polymerase chain reaction (qPCR).@*RESULTS@#Masson's staining result showed fewer pseudolobule structures and fibrous connective tissue in the GSG-treatment groups than in the spontaneous recovery group. Ultrasonography showed that GSG treatment reduced the number of punctate hyperechoic lesions in mice cirrhotic livers. The serum ALT, AST, HA levels were significantly ameliorated by GSG treatment (ALT: F=8.104, P=0.000; AST: F=7.078, P=0.002; and HA: F=7.621, P=0.001). The expression levels of collagen-I and SMA in the cirrhotic livers were also attenuated by GSG treatment (collagen-I: F=3.938, P=0.011; SMA: F=4.115, P=0.009). Tregs, which were elevated in the fibrotic livers, were suppressed by GSG treatment (F=8.268, P=0.001). The expressions of IL-6, TNF-α and IL-1β increased, and TGF-β levels decreased in the cirrhotic livers after GSG treatment (IL-6: F=5.457, P=0.004; TNF-α: F=6.023, P=0.002; IL-1β: F=6.658, P=0.001; and TGF-β1: F=11.239, P=0.000).@*CONCLUSIONS@#GSG promoted the resolution/regression of cirrhosis and restored liver functions in part by suppressing Treg cell differentiation, which may be mediated by hepatic stellate cells.


Asunto(s)
Animales , Masculino , Ratones , Modelos Animales de Enfermedad , Medicamentos Herbarios Chinos , Farmacología , Usos Terapéuticos , Células Estrelladas Hepáticas , Cirrosis Hepática Experimental , Quimioterapia , Alergia e Inmunología , Ratones Endogámicos BALB C , Linfocitos T Reguladores
2.
Chinese Journal of Experimental and Clinical Virology ; (6): 250-252, 2013.
Artículo en Chino | WPRIM | ID: wpr-318051

RESUMEN

<p><b>OBJECTIVE</b>To study whether CO-Q10 can protect liver injury caused by acute on chronic liver failure (ACLF) by autophagy.</p><p><b>METHODS</b>Rats were separated into three groups: control group, acute on chronic liver failure (ACLF) and intervenient group, liver tissues were observed by optical microscopy and electron microscopy. The levels of Beclin-1 expression were determined by real-time PCR. And Western Blot.</p><p><b>RESULTS</b>Areas of necrosis detected in intervenient group were alleviated than in ACLF significantly. Most mitochondrias had been degradated in ACLF group while alive in intervenient group. Real-time PCR and Western Blot revealed level of beclin-1 in ACLF was lower than control and intervenient group.</p><p><b>CONCLUSION</b>Intervenient group may ameliorate rat liver injury by promoting autophagy.</p>


Asunto(s)
Animales , Humanos , Masculino , Ratas , Proteínas Reguladoras de la Apoptosis , Genética , Metabolismo , Autofagia , Beclina-1 , Fallo Hepático Agudo , Genética , Metabolismo , Mitocondrias Hepáticas , Metabolismo , Ratas Sprague-Dawley , Ubiquinona , Metabolismo
3.
Chinese Journal of Hepatology ; (12): 693-697, 2012.
Artículo en Chino | WPRIM | ID: wpr-296826

RESUMEN

<p><b>OBJECTIVE</b>To determine the mechanism underlying the therapeutic activities of glycogen synthase kinase 3b (GSK3b) against hepatic ischemia-reperfusion (H-IR) injury by investigating the inhibitive effects of GSK3b on inflammation mediated by Toll-like receptor 4 (TLR4).</p><p><b>METHODS</b>C57BL/6 male mice were subjected to 90 min of warm liver cephalad lobe ischemia, followed by reperfusion for various lengths of time. The mice were divided into three groups: the H-IR untreated model (control group), and the H-IR inflammation-induced models that received an intraperitoneal injection of purified lipopolysaccharide (LPS) endotoxin alone (inflammation group) or with pretreatment of the SB216763 GSK3b-specific inhibitor (intervention group). To create a parallel isolated cell system for detailed investigations of macrophages, marrow-derived stem cells were isolated from femurs of the H-IR control group of mice and used to derive primary macrophages. The cells were then divided into the same three groups as the whole mouse system: control, LPS-induced inflammation model, and inflammation model with SB216763 intervention. Differential expressions of inflammation-related proteins and genes were detected by Western blotting and real-time quantitative PCR, respectively.</p><p><b>RESULTS</b>The phosphorylation levels of ERK, JNK and p38 MAPK were induced in liver at 1 h after reperfusion, but then steadily decreased and returned to baseline levels by 4 h after reperfusion. In addition, the phosphorylation levels of ERK and JNK were induced in macrophages at 15 min after LPS stimulation, while the phosphorylation level of p38 MAPK was induced at 1 h; SB216763 pretreatment suppressed the LPS-stimulated ERK, JNK and p38 phosphorylation in macrophages. In the mouse model, GSK3b activity was found to promote the gene expression of anti-inflammatory cytokine IL-10 (control: 0.21 ± 0.08, inflammation: 0.83 ± 0.21, intervention: 1.76 ± 0.67; F = 3.16, P = 0.027) but to significantly inhibit the gene expression of pro-inflammatory cytokines IL-12 (control: 0.11 ± 0.05, inflammation: 0.85 ± 0.11, intervention: 0.43 ± 0.10; F = 2.67, P = 0.038), TNF-a, (control: 0.052 ± 0.012, inflammation: 8.11 ± 0.98, intervention: 3.9 ± 0.82; F = 4.13, P = 0.016), IL-6 (control: 0.22 ± 0.08, inflammation: 6.37 ± 0.81, intervention: 2.11 ± 0.63; F = 3.21, P = 0.024), and IL-1b (control: 0.12 ± 0.07, inflammation: 2.51 ± 0.62, and intervention: 1.28 ± 0.33; F = 2.22, P = 0.030).</p><p><b>CONCLUSION</b>Inhibition of GSK3b selectively regulates the expression of anti-inflammatory and pro-inflammatory cytokines in liver Kupffer cells (liver macrophages). This process may be one of the mechanisms underlying the ability of GSK3b to ameliorate hepatic ischemia-reperfusion injury, possibly because inhibition of pro-inflammatory cytokines may indirectly mediate liver cell apoptosis.</p>


Asunto(s)
Animales , Masculino , Ratones , Células Cultivadas , Citocinas , Metabolismo , Glucógeno Sintasa Quinasa 3 , Metabolismo , Glucógeno Sintasa Quinasa 3 beta , Inflamación , Metabolismo , Patología , Lipopolisacáridos , Hígado , Patología , Macrófagos , Metabolismo , Ratones Endogámicos C57BL , Daño por Reperfusión , Receptor Toll-Like 4 , Metabolismo
4.
Chinese Journal of Experimental and Clinical Virology ; (6): 275-277, 2009.
Artículo en Chino | WPRIM | ID: wpr-325568

RESUMEN

<p><b>OBJECTIVE</b>To comprehend the latest HIV-I epidemic tendency and the character of V3 loop in MSM population of Beijing. METHODS; The C2-V3 regions of the HIV envelop gene were amplified by nest-PCR and sequenced from 11 HIV-l-infected MSM in Beijing in 2007. The subtype and sequences of V3 loop was analyzed. RESULTS There are 4 subtype B strains, 5 CRF AE, 1 CRFO7BC and 1 CRF15-01B strains within all 11 strains. There are five types of central motifs of the 11 samples, in which GPGR and GPGQ are most common.</p><p><b>CONCLUSION</b>Recombination subtype of HIV-1 are spread extensively in MSM population of Beijing.</p>


Asunto(s)
Adulto , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Secuencia de Aminoácidos , China , Infecciones por VIH , Virología , VIH-1 , Química , Clasificación , Genética , Homosexualidad Masculina , Datos de Secuencia Molecular , Filogenia , Alineación de Secuencia , Productos del Gen env del Virus de la Inmunodeficiencia Humana , Química , Genética
5.
Chinese Journal of Experimental and Clinical Virology ; (6): 446-448, 2009.
Artículo en Chino | WPRIM | ID: wpr-325516

RESUMEN

<p><b>OBJECTIVE</b>To investigate alterations of hyper variable region 1 (HR 1) of mitochondrial DNA Blood cells were (mtDNA) in white blood cells of Chinese Han nationality HIV/AIDS patients.</p><p><b>METHODS</b>obtained from 47 cases of therapy-naïve HIV/AIDS patients without opportunity infection and DNA were extracted using blood DNA extracted kit. About 600 bp fragments which contain HR 1 were amplified by PCR. Alterations were determined by directed DNA sequencing.</p><p><b>RESULTS</b>There were 124 polymorphism sites in mtDNA HR 1 (nb16024-16383) in 47 HIV/AIDS patients. The alteration rate was 0 to 20.47% (median 5.33%). 36 cases experienced C to T nucleotide change at nt 16 223, and the alteration rate was 70.97%. At nt 16 362, 26 individuals showed T to C nucleotide change and 3 individuals showed T to G alteration, alteration rate was 55.32% (26/47) and 6.38% (3/47) individually.</p><p><b>CONCLUSION</b>HIV infection may cause more alterations in HR 1 regions.</p>


Asunto(s)
Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pueblo Asiatico , Etnología , Genética , Secuencia de Bases , Células Cultivadas , China , ADN Mitocondrial , Química , Genética , Variación Genética , Infecciones por VIH , Etnología , Genética , Leucocitos , Química , Datos de Secuencia Molecular , Mutación , Polimorfismo Genético , Homología de Secuencia de Ácido Nucleico
6.
Chinese Journal of Experimental and Clinical Virology ; (6): 27-29, 2008.
Artículo en Chino | WPRIM | ID: wpr-254151

RESUMEN

<p><b>OBJECTIVE</b>To characterize the variation in V3 loop of HIV-1 B'strains circulating in Chinese blood donors.</p><p><b>METHODS</b>The c2-c3 regions of the HIV envelop gene were amplified by nest-PCR from 32 HIV-1-infected blood donors in He Nan province in China. The BIOEDIT and MEGA software are used to analyze the sequences of V3 loop.</p><p><b>RESULTS</b>There are five types of central motifs of the 32 samples, in which GPGR and GPGQ are most common. More variations associated with T tropic/SI phenotype can be seen in AIDS group.</p><p><b>CONCLUSION</b>The V3 tip motifs of HIV-1B' strains circulating in Chinese blood donors are various, the different characterization of V3 loop between AIDS and asymptomatic patients indicates different biological phenotype and pathogenesis which warrant additional investigation.</p>


Asunto(s)
Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Donantes de Sangre , China , Variación Genética , Genética , Proteína gp120 de Envoltorio del VIH , Química , Genética , Infecciones por VIH , Sangre , Virología , VIH-1 , Química , Clasificación , Genética , Datos de Secuencia Molecular , Oligopéptidos , Química , Fenotipo
7.
Chinese Journal of Experimental and Clinical Virology ; (6): 186-188, 2008.
Artículo en Chino | WPRIM | ID: wpr-254109

RESUMEN

<p><b>OBJECTIVE</b>To investigate the regulatory effect and significance of transcription factor E2F1 on X-ray repair cross2 complementing 1 (XRCC1).</p><p><b>METHODS</b>Saos2 cells were transfected with the E2F1 expression vectors (tet-E2F1) and mutated E2F1 expression vectors (tet-132E). XRCC1 promotor luciferase reporter vector was constructed and transfected into Saos2 cells together with E2F1, E2F2, E2F3 and E2F4 expression vectors at different amount. The cells were collected 36 hours post-transfection for luciferase assays and absorbance was read at 570 nm.</p><p><b>RESULTS</b>Cotransfection of increasing amounts of E2F1 expression vector with the XRCC1 promoter-luciferase reporter caused a dose-dependent increase in luciferase activation. In contrast, DNA binding incompetent E2F1 (132E) could not activate the XRCC1 promoter-luciferase reporter.</p><p><b>CONCLUSION</b>E2F1 could upregulate endogenous XRCC1 expression and stimulate the XRCC1 promoter.</p>


Asunto(s)
Humanos , Línea Celular Tumoral , Proteínas de Unión al ADN , Genética , Metabolismo , Factor de Transcripción E2F1 , Genética , Metabolismo , Expresión Génica , Genes Reporteros , Regiones Promotoras Genéticas , Unión Proteica , Regulación hacia Arriba , Proteína 1 de Reparación por Escisión del Grupo de Complementación Cruzada de las Lesiones por Rayos X
8.
Chinese Journal of Experimental and Clinical Virology ; (6): 287-289, 2008.
Artículo en Chino | WPRIM | ID: wpr-254079

RESUMEN

<p><b>OBJECTIVE</b>To highly express TAT-HBX-EGFP fusion protein and study its distribution in mouse liver.</p><p><b>METHODS</b>TAT-HBX-EGFP recombinant vector was constructed and fusion protein was induced by IPTG and expression in BL21; fusion protein was purified by Ni-NTA argarose, then injected into the peritoneal cavity of the mice. Distribution of fusion protein was observed by immunofluorescence.</p><p><b>RESULTS</b>TAT-HBX-EGFP was highly expression in E. coli; HBX could be induced into mouse liver by TAT.</p><p><b>CONCLUSION</b>HBX protein could be induced into mouse liver by TAT induced peptide.</p>


Asunto(s)
Animales , Femenino , Humanos , Masculino , Ratones , Membrana Celular , Genética , Metabolismo , Escherichia coli , Genética , Metabolismo , Expresión Génica , Proteínas Fluorescentes Verdes , Genética , Metabolismo , Hepatitis B , Metabolismo , Virología , Hígado , Metabolismo , Ratones Endogámicos ICR , Transporte de Proteínas , Proteínas Recombinantes de Fusión , Genética , Metabolismo , Transactivadores , Genética , Metabolismo , Proteínas Reguladoras y Accesorias Virales , Genética , Metabolismo , Productos del Gen tat del Virus de la Inmunodeficiencia Humana , Genética , Metabolismo
9.
Chinese Journal of Experimental and Clinical Virology ; (6): 220-222, 2007.
Artículo en Chino | WPRIM | ID: wpr-248798

RESUMEN

<p><b>OBJECTIVE</b>To investigate the expression of K18, Ser-33 and Ser-52 phosphorylated K18 in HBV infected human liver disease and its significance.</p><p><b>METHODS</b>The expression and localization of K18 and Ser-33, Ser-52 phosphorylated K18 in healthy liver tissue, in liver tissues of patients with post-HBV infection cirrhosis and severe chronic hepatitis were detected by histochemistry.</p><p><b>RESULTS</b>K18, Ser-33 and Ser-52 phosphorylated K18 were expressed in normal liver cells, in liver tissues of cirrhosis patients and severe chronic hepatitis cases. The expression of K18 in the liver cells from the 3 different sources had no significant difference in levels. Ser-33 and Ser-52 phosphorylated K18 were expressed in normal liver cells, in liver tissues of cirrhosis patients chronicity HBV hepatitis and severe chronic hepatitis cases. Ser-33 and Ser-52 located around cytoplasmic membrane, diffused into cytoplasm and expressed at a higher levels in cirrhosis and severe chronic hepatitis.</p><p><b>CONCLUSION</b>The expression levels of Ser-33 and Ser-52 phosphorylated K18 increased along with the progression of HBV infected human liver disease. The phosphorylation of K18 could be a marker of progression of HBV infected human liver disease.</p>


Asunto(s)
Humanos , Hepatitis B , Metabolismo , Inmunohistoquímica , Queratina-18 , Metabolismo , Cirrosis Hepática , Metabolismo , Patología , Virología , Hepatopatías , Metabolismo , Patología , Virología , Fosforilación , Serina , Metabolismo
10.
Acta Academiae Medicinae Sinicae ; (6): 642-646, 2006.
Artículo en Chino | WPRIM | ID: wpr-313716

RESUMEN

<p><b>OBJECTIVE</b>To clone human CCL3L1 cDNA and to express and purify the glutathione-S-transferase (GST) fusion protein and human CCL3L1 protein.</p><p><b>METHODS</b>Total RNA was isolated from breast cancer cell line MCF7. CCL3L1 cDNA including open reading frame was obtained by RT-PCR. PCR product was digested with EcoR I and cloned into the pGEX-4T-1 vector. The plasmids from positive clone was prepared and sequenced to confirm the CCL3L1 in correct fusion form. pGEX-4T-CCL3L1 was transfected to BL21 E. coli via isopropyl-beta-D-thiogalactoside (IPTG) induction to produce GST-CCL3L1 fusion protein, which was further detected by SDS-PAGE and Western blotting.</p><p><b>RESULTS</b>As shown and confirmed by restriction endonuclease digestion analysis, CCL3L1 was correctly inserted into pGEX-4T-1 vector. The expressed fusion protein had a relative molecular weight of approximately 34 kD.</p><p><b>CONCLUSION</b>GST-CCL3L1 fusion protein can be successfully expressed using appropriate vector.</p>


Asunto(s)
Femenino , Humanos , Línea Celular Tumoral , Quimiocinas CC , Genética , Clonación Molecular , ADN Complementario , Genética , Escherichia coli , Genética , Metabolismo , Vectores Genéticos , Glutatión Transferasa , Genética , Isopropil Tiogalactósido , Farmacología , Proteínas Recombinantes de Fusión , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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