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1.
Adv Sci (Weinh) ; 5(10): 1800873, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30356983

RESUMO

Biomineralization in vertebrates is initiated via amorphous calcium phosphate (ACP) precursors. These precursors infiltrate the extracellular collagen matrix where they undergo phase transformation into intrafibrillar carbonated apatite. Although it is well established that ACP precursors are released from intracellular vesicles through exocytosis, an unsolved enigma in this cell-mediated mineralization process is how ACP precursors, initially produced in the mitochondria, are translocated to the intracellular vesicles. The present study proposes that mitophagy provides the mechanism for transfer of ACP precursors from the dysfunctioned mitochondria to autophagosomes, which, upon fusion with lysosomes, become autolysosomes where the mitochondrial ACP precursors coalesce to form larger intravesicular granules, prior to their release into the extracellular matrix. Apart from endowing the mitochondria with the function of ACP delivery through mitophagy, the present results indicate that mitophagy, triggered upon intramitochondrial ACP accumulation in osteogenic lineage-committed mesenchymal stem cells, participates in the biomineralization process through the BMP/Smad signaling pathway.

2.
Cell Death Discov ; 4: 32, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30245855

RESUMO

Photoreceptor cell death is recognized as the key pathogenesis of retinal degeneration, but the molecular basis underlying photoreceptor-specific cell loss in retinal damaging conditions is virtually unknown. The N-myc downstream regulated gene (NDRG) family has recently been reported to regulate cell viability, in particular NDRG1 has been uncovered expression in photoreceptor cells. Accordingly, we herein examined the potential roles of NDRGs in mediating photoreceptor-specific cell loss in retinal damages. By using mouse models of retinal degeneration and the 661 W photoreceptor cell line, we showed that photoreceptor cells are indeed highly sensitive to light exposure and the related oxidative stress, and that photoreceptor cells are even selectively diminished by phototoxins of the alkylating agent N-Methyl-N-nitrosourea (MNU). Unexpectedly, we discovered that of all the NDRG family members, NDRG2, but not the originally hypothesized NDRG1 or other NDRG subtypes, was selectively expressed and specifically responded to retinal damaging conditions in photoreceptor cells. Furthermore, functional experiments proved that NDRG2 was essential for photoreceptor cell viability, which could be attributed to NDRG2 control of the photo-oxidative stress, and that it was the suppression of NDRG2 which led to photoreceptor cell loss in damaging conditions. More importantly, NDRG2 preservation contributed to photoreceptor-specific cell maintenance and retinal protection both in vitro and in vivo. Our findings revealed a previously unrecognized role of NDRG2 in mediating photoreceptor cell homeostasis and established for the first time the molecular hallmark of photoreceptor-specific cell death as NDRG2 suppression, shedding light on improved understanding and therapy of retinal degeneration.

3.
Nan Fang Yi Ke Da Xue Xue Bao ; 38(3): 268-273, 2018 Mar 20.
Artigo em Chinês | MEDLINE | ID: mdl-29643031

RESUMO

OBJECTIVE: To investigate the role of p38MAPK signaling pathway in autophagy of intestinal epithelial cells induced by spvB of S.typhimurium. METHODS: Henle-407 cells in exponential growth were infected with wild-type S.typhimurium strain STM-211 (with spvB gene), spvB mutated strain STM-delata;spvB, or with delata;spvB-complemented strain STM-c-spvB after treatment of the cells with the p38MAPK inhibitor SB203580. At different time points of co-culture, the cells were collected and the intracellular bacteria were counted. Western blotting was performed to detect the expressions of phosphorylated p38 and autophagy-related proteins LC3 and p62; immunofluorescence staining was used to observe the expression and distribution of LC3. RESULTS: At 1, 2 and 4 h after the infection, the phosphorylation levels of p38 in STM-211 group and STM-c-spvB group were significantly lower than that in STM-delata;spvB group (P<0.05). At 2 and 4 h of co-culture, the intracellular bacterial counts were significantly greater in STM-211 and STM-c-spvB infection groups than in STM-delata;spvB group (P<0.05). Pretreatment with p38 inhibitor SB203580 did no significantly affect the expression levels of LC3 II or P62 in STM-211 and STM-c-spvB groups, but caused significant reduction in their expressions in STM-delata;spvB group at 1 h (P<0.05), and such changes were more obvious at 3 h (P<0.05). CONCLUSION: The inhibitory effect of spvB gene on autophagy in intestinal epithelial cells is related with the negative regulation of p38MAPK signaling pathway.


Assuntos
ADP Ribose Transferases/genética , Autofagia , Células Epiteliais/citologia , Sistema de Sinalização das MAP Quinases , Salmonella typhimurium , Fatores de Virulência/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Células HeLa , Humanos , Intestinos/citologia , Salmonella typhimurium/genética , Transdução de Sinais , Virulência
4.
IDCases ; 9: 21-24, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28560174

RESUMO

To define the clinical manifestations and laboratory characteristics of pediatric severe fever with thrombocytopenia syndrome (SFTS) case caused by a novel bunyavirus. we retrospectively analyzed a pediatric case of viral SFTS in a 13 year old successfully managed and confirmed to be due to the novel bunyavirus now referred to as Huaiyangshan virus. A literature review of related cases was performed.Our pediatric case was a 13.3-year-old middle school student no underlying disease. Major clinical features included a fever with chills, headache, and dizziness. The patient's epidemiology showed he had close contact with his grandfather who had a confirmed, novel bunyavirus infection. Symptomatic theraphy were given at admission. The patient's temperature and platelet count returned to normal by days 7 and 10, respectively, and he was discharged from the hospital with an improved condition. A literature search was performed using "severe fever with thrombocytopenia syndrome" and "bunyavirus" as keywords, but few relevant reports were found. Novel bunyavirus infection can be transmitted through close contact. Confirmed cases should be kept in isolation. Clinical manifestations were characterized by aspecific symptoms, such as fever and chills. In some cases, platelet counts may remain normal in the early phase of the disease, and fever may not present throughout the entire illness period. Thus, misdiagnosis is possible. Surveillance and vigorous follow-up should be carried out in children with tick bites or in close contact with an index patient in high-risk areas during peak season.

5.
Molecules ; 22(5)2017 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-28534823

RESUMO

Two novel glucosylated zinc(ІІ) phthalocyanines 7a-7b, as well as the acetyl-protected counterparts 6a-6b, have been synthesized by the Cu(I)-catalyzed 1,3-dipolar cycloaddition between the propargylated phthalocyanine and azide-substituted glucoses. All of these phthalocyanines were characterized with various spectroscopic methods and studied for their photo-physical, photo-chemical, and photo-biological properties. With glucose as the targeting unit, phthalocyanines 7a-7b exhibit a specific affinity to MCF-7 breast cancer cells over human embryonic lung fibroblast (HELF) cells, showing higher cellular uptake. Upon illumination, both photosensitizers show high cytotoxicity with IC50 as low as 0.032 µM toward MCF-7 cells, which are attributed to their high cellular uptake and low aggregation tendency in the biological media, promoting the generation of intracellular reactive oxygen species (ROS). Confocal laser fluorescence microscopic studies have also revealed that they have high and selective affinities to the lysosomes, but not the mitochondria, of MCF-7 cells. The results show that these two glucosylated zinc(II) phthalocyanines are potential anticancer agents for targeting photodynamic therapy.


Assuntos
Indóis/síntese química , Compostos Organometálicos/síntese química , Fármacos Fotossensibilizantes/síntese química , Espécies Reativas de Oxigênio/agonistas , Zinco/química , Acetilação , Cátions Bivalentes , Linhagem Celular , Reação de Cicloadição , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Glucose/química , Glicosilação , Humanos , Indóis/farmacologia , Concentração Inibidora 50 , Luz , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Células MCF-7 , Especificidade de Órgãos , Compostos Organometálicos/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Espécies Reativas de Oxigênio/metabolismo
6.
J Huazhong Univ Sci Technolog Med Sci ; 37(1): 105-109, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28224437

RESUMO

Hepatitis associated anti-tuberculous treatment (HATT) has been a main obstacle in managing patients co-infected with Mycobacterium tuberculosis and hepatitis B virus (HBV). Therefore, we evaluated the factors related to the severity of adverse effects during HATT, especially those associated with liver failure. A retrospective study was carried out at Tongji Hospital from 2007 to 2012. Increases in serum transaminase levels of >3, 5, and 10 times the upper limit of normal (ULN) were used to define liver damage as mild, moderate, and severe, respectively. Patients with elevated total bilirubin (TBil) levels that were more than 10 times the ULN (>171 µmol/L) with or without decreased (<40%) prothrombin activity (PTA) were diagnosed with liver failure. A cohort of 87 patients was analyzed. The incidence of liver damage and liver failure was 59.8% (n=52) and 25.3% (n=22), respectively. The following variables were correlated with the severity of hepatotoxicity: albumin (ALB) levels, PTA, platelet counts (PLT), and the use of antiretroviral therapies (P<0.05). Hypo-proteinemia and antiretroviral therapy were significantly associated with liver failure, and high viral loads were a significant risk factor with an odds ratio (OR) of 2.066. Judicious follow-up of clinical conditions, liver function tests, and coagulation function, especially in patients with high HBV loads and hypoalbuminemia is recommended. It may be advisable to reconsider the use of antiviral drugs failure during the course of anti-tuberculous treatment of HBV infection patients to avoid the occurrence of furious liver failure.


Assuntos
Antituberculosos/efeitos adversos , Doença Hepática Induzida por Substâncias e Drogas/epidemiologia , Hepatite B Crônica/etiologia , Falência Hepática/epidemiologia , Tuberculose/tratamento farmacológico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença Hepática Induzida por Substâncias e Drogas/virologia , Feminino , Hepatite B Crônica/epidemiologia , Hepatite B Crônica/fisiopatologia , Humanos , Incidência , Falência Hepática/induzido quimicamente , Falência Hepática/virologia , Testes de Função Hepática , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Exacerbação dos Sintomas , Transaminases/sangue , Tuberculose/metabolismo , Tuberculose/fisiopatologia , Carga Viral , Adulto Jovem
7.
Arch Virol ; 161(10): 2667-72, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27383210

RESUMO

Equine infectious anemia virus (EIAV) is a member of the genus Lentivirus of the family Retroviridae. Horses are the most susceptible equids to EIAV infection and are therefore the primary hosts of this virus. In contrast, infected donkeys do not develop clinically active equine infectious anemia (EIA). This phenomenon is similar to what has been observed with HIV-1, which fails to induce AIDS in non-human primates. Interestingly, Shen et al. developed a donkey-tropic pathogenic virus strain (EIAVDV117, DV117) by serially passaging a horse-tropic pathogenic strain, EIAVLN40 (LN40), in donkeys. LN40, which was generated by passaging a field isolate in horses, displayed enhanced virulence in horses but caused no clinical symptoms in donkeys. Infection with DV117 induced acute EIA in nearly 100 % of donkeys. Genomic analysis of DV117 revealed a significantly higher frequency of A-to-G substitutions when compared to LN40. Furthermore, detailed analysis of dinucleotide editing showed that A-to-G mutations had a preference for 5'TpA and 5'ApA. These results strongly implicated the activity of the adenosine deaminase, ADAR1, in this type of mutation. Further investigation demonstrated that overexpression of donkey ADAR1 increased A-to-G mutations within the genome of EIAV. Together with our previous finding that multiple mutations in multiple genes are generated in DV117 during its adaptation from horses to donkeys, the present study suggests that ADAR1-induced A-to-G mutations occur during virus adaption to related new hosts contributing to the alteration of EIAV host tropism.


Assuntos
Adaptação Biológica , Adenosina Desaminase/metabolismo , Vírus da Anemia Infecciosa Equina/genética , Vírus da Anemia Infecciosa Equina/patogenicidade , RNA de Cadeia Dupla/metabolismo , Animais , Equidae , Cavalos , Mutação Puntual , Análise de Sequência de DNA , Inoculações Seriadas
8.
Int Immunopharmacol ; 36: 86-93, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27107801

RESUMO

Periodontitis is a severe inflammatory response, leading to characteristic periodontal soft tissue destruction and alveolar bone resorption. Baicalin possesses potent anti-inflammatory activity; however, it is still unclear whether baicalin regulates toll-like receptor (TLR) 2/4 expression and downstream signaling during the process of periodontitis. In this study, the cervical area of the maxillary second molars of rats was ligated and inoculated with Porphyromonas gingivalis (P. gingivalis) for 4weeks to induce periodontitis. Some rats with periodontitis were treated intragastrically with baicalin (50, 100 or 200mg/kg/day) or vehicle for 4weeks. Compared with the sham group, the levels of TLR2, TLR4 and MyD88 expression and the p38 MAPK and NF-κB activation were up-regulated in the experimental periodontitis group (EPG), accompanied by marked alveolar bone loss and severe inflammation. Treatment with 100 or 200mg/kg/day baicalin dramatically reduced the alveolar bone loss, the levels of HMGB1, TNF-α, IL-1ß, and MPO expression, and the numbers of inflammatory infiltrates in the gingival tissues. Importantly, treatment with 100 or 200mg/kg/day baicalin mitigated the periodontitis-up-regulated TLR2, TLR4 and MyD88 expression, and the p38 MAPK and NF-κB activation. Hence, the blockage of the TLR2 and TLR4/MyD88/p38 MAPK/NF-κB signaling by baicalin may contribute to its anti-inflammatory effects in rat model of periodontitis. In conclusion, these novel findings indicate that baicalin inhibits the TLR2 and TLR4 expression and the downstream signaling and mitigates inflammatory responses and the alveolar bone loss in rat experimental periodontitis. Therefore, baicalin may be a potential therapeutic agent for treatment of periodontitis.


Assuntos
Anti-Inflamatórios/uso terapêutico , Infecções por Bacteroidaceae/tratamento farmacológico , Flavonoides/uso terapêutico , Fator 88 de Diferenciação Mieloide/metabolismo , Periodontite/tratamento farmacológico , Porphyromonas gingivalis/fisiologia , Scutellaria baicalensis/imunologia , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Infecções por Bacteroidaceae/imunologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Modelos Animais , Fator 88 de Diferenciação Mieloide/genética , Periodontite/imunologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Receptor 2 Toll-Like/genética , Receptor 4 Toll-Like/genética
9.
Virology ; 476: 364-371, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25589239

RESUMO

Adenosine deaminases that act on RNA (ADARs) have been reported to be functional on various viruses. ADAR1 may exhibit antiviral or proviral activity depending on the type of virus. Human immunodeficiency virus (HIV)-1 is the most well-studied lentivirus with respect to its interaction with ADAR1, and variable results have been reported. In this study, we demonstrated that equine ADAR1 (eADAR1) was a positive regulator of equine infectious anemia virus (EIAV), another lentivirus of the Retroviridae family. First, eADAR1 significantly promoted EIAV replication, and the enhancement of viral protein expression was associated with the long terminal repeat (LTR) and Rev response element (RRE) regions. Second, the RNA binding domain 1 of eADAR1 was essential only for enhancing LTR-mediated gene expression. Third, in contrast with APOBEC proteins, which have been shown to reduce lentiviral infectivity, eADAR1 increased the EIAV infectivity. This study indicated that eADAR1 was proviral rather than antiviral for EIAV.


Assuntos
Adenosina Desaminase/metabolismo , Anemia Infecciosa Equina/enzimologia , Vírus da Anemia Infecciosa Equina/fisiologia , Vírus da Anemia Infecciosa Equina/patogenicidade , RNA de Cadeia Dupla/metabolismo , RNA Viral/metabolismo , Replicação Viral , Adenosina Desaminase/genética , Animais , Linhagem Celular , Anemia Infecciosa Equina/genética , Anemia Infecciosa Equina/virologia , Cavalos , Interações Hospedeiro-Patógeno , Vírus da Anemia Infecciosa Equina/genética , Estrutura Terciária de Proteína , RNA de Cadeia Dupla/genética , RNA Viral/química , RNA Viral/genética , Sequências Repetidas Terminais , Virulência
10.
J Virol ; 88(21): 12296-310, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25122784

RESUMO

UNLABELLED: Viperin is an endoplasmic reticulum (ER)-associated multifunctional protein that regulates virus replication and possesses broad antiviral activity. In many cases, viperin interferes with the trafficking and budding of viral structural proteins by distorting the membrane transportation system. The lentivirus equine infectious anemia virus (EIAV) has been studied extensively. In this study, we examined the restrictive effect of equine viperin (eViperin) on EIAV replication and investigated the possible molecular basis of this restriction to obtain insights into the effect of this cellular factor on retroviruses. We demonstrated that EIAV infection of primary equine monocyte-derived macrophages (eMDMs) upregulated the expression of eViperin. The overexpression of eViperin significantly inhibited the replication of EIAV in eMDMs, and knockdown of eViperin transcription enhanced the replication of EIAV in eMDMs by approximately 45.8%. Further experiments indicated that eViperin restricts EIAV at multiple steps of viral replication. The overexpression of eViperin inhibited EIAV Gag release. Both the α-helix domain and radical S-adenosylmethionine (SAM) domain were required for this activity. However, the essential motifs in SAM were different from those reported for the inhibition of HIV-1 Gag by human viperin. Furthermore, eViperin disrupted the synthesis of both EIAV Env and receptor, which consequently inhibited viral production and entry, respectively, and this disruption was dependent on the eViperin α-helix domain. Using immunofluorescence assays and electron microscopy, we demonstrated that the α-helix domain is responsible for the distortion of the endoplasmic reticulum (ER). Finally, EIAV did not exhibit counteracting eViperin at the protein level. IMPORTANCE: In previous studies, viperin was indicated as restricting virus replications primarily by the inhibition of virus budding. Here, we show that viperin may have multiple antiviral mechanisms, including the reduction of EIAV Gag budding and Env expression, and these activities are dependent on different viperin domains. We especially demonstrate that the overexpression of viperin inhibits EIAV entry by decreasing the level of virus receptor. Therefore, viperin restriction of viruses is determined largely by the dependence of virus on the cellular membrane transportation system.


Assuntos
Retículo Endoplasmático/metabolismo , Retículo Endoplasmático/virologia , Interações Hospedeiro-Patógeno , Vírus da Anemia Infecciosa Equina/imunologia , Vírus da Anemia Infecciosa Equina/fisiologia , Proteínas Virais/metabolismo , Replicação Viral , Animais , Células Cultivadas , Retículo Endoplasmático/ultraestrutura , Imunofluorescência , HIV-1 , Cavalos , Macrófagos/imunologia , Macrófagos/virologia , Microscopia Eletrônica , Liberação de Vírus
11.
Zhonghua Nan Ke Xue ; 19(7): 630-3, 2013 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-23926681

RESUMO

OBJECTIVE: To observe the clinical effect of low-dose testosterone undecanoate capsules combined with tadalafil on late-onset hypogonadism (LOH) accompanied with ED. METHODS: Ninety cases of LOH accompanied with ED who met the inclusion criteria were randomly divided into a control group and a combination therapy group, the former treated with tadalafil and the latter with low-dose testosterone undecanoate capsules combined with tadalafil. The LOH symptoms, IIEF-5 scores, sexual encounter profile (SEP) scores, prostate volumes, and the levels of total testosterone (TT), free testosterone (FT) and prostatic specific antigen (PSA) were recorded and compared between the two groups before and after treatment. RESULTS: The IIEF-5 and SEP scores and the TT and FT levels were 20.6 +/- 3.8, 4.02 +/- 1.08, (15.4 +/- 3.4) nmol/L and (0.391 +/- 0.062) nmol/L, respectively, in the combination therapy group after treatment, significantly higher both than 15.7 +/- 3.9, 1.49 +/- 0.82, (10.1 +/- 1.2) nmol/L and (0.200 +/- 0.045) nmol/L before treatment (P < 0.01) and than 8.6 +/- 3.6, 3.50 +/- 1.21, (10.2 +/- 1.2) nmol/L and (0.210 +/- 0.051) nmol/L in the control group after treatment (P < 0.01). CONCLUSION: Low-dose testosterone undecanoate capsules combined with tadalafil has a definite clinical effect and no obvious adverse reactions in the treatment of LOH accompanied with ED.


Assuntos
Carbolinas/uso terapêutico , Disfunção Erétil/tratamento farmacológico , Hipogonadismo/tratamento farmacológico , Testosterona/análogos & derivados , Adulto , Carbolinas/administração & dosagem , Quimioterapia Combinada , Disfunção Erétil/complicações , Humanos , Hipogonadismo/complicações , Masculino , Pessoa de Meia-Idade , Tadalafila , Testosterona/administração & dosagem , Testosterona/uso terapêutico , Resultado do Tratamento
12.
Zhonghua Nan Ke Xue ; 19(3): 241-6, 2013 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-23700731

RESUMO

OBJECTIVE: To investigate the effect of low-dose daily de-escalatory administration of tadalafil on psychological erectile dysfunction (ED). METHODS: We randomized 84 psychological ED patients into an observation and a control group of equal number to receive low-dose daily de-escalatory administration and on-demand medication of tadalafil, respectively, both for 2 months. We compared the scores on IIEF-5 and erection hardness (EHS) between the two groups before and after the treatment. RESULTS: The treatment and follow-up were accomplished for 79 cases, with 5 withdrawals in the control group. The IIEF-5 and EHS scores were remarkably improved in both the observation and control groups after treatment. The rate of therapeutic effectiveness was significantly higher in the observation group than in the control (95.2% vs 86.5%, P < 0.05). CONCLUSION: Low-dose daily de-escalatory administration of tadalafil is highly effective and even better than on-demand medication of tadalafil for psychological ED.


Assuntos
Carbolinas/administração & dosagem , Disfunção Erétil/tratamento farmacológico , Inibidores de Fosfodiesterase/administração & dosagem , Adulto , Carbolinas/uso terapêutico , Disfunção Erétil/psicologia , Humanos , Masculino , Inibidores de Fosfodiesterase/uso terapêutico , Tadalafila , Adulto Jovem
13.
Shanghai Kou Qiang Yi Xue ; 22(6): 634-42, 2013 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-24469124

RESUMO

PURPOSE: The purpose of this study was to culture and identify dental pulp stem cells(DPSCs) from deciduous teeth in vitro and construct the recombinant hFOXA2 and hPDX1 lentivirus vectors and transfect the DPSCs to induce insulin-producing cells (IPCs). METHODS: DPSCs were separated and cultured by enzyme digest method, and purified by limited dilution method. Flow cytometry was used to determine the surface marker expression of the DPSCs, and the ability of multiple differentiations was determined by specific staining. hFOXA2 and hPDX1 genes were amplified by PCR, and the recombinant hFOXA2 and hPDX1 lentivirus vectors were reconstructed and transfected into 293T cells by lipofectamine2000 for virus packaging. The viral infection efficiency and titer were determined through fluorescence cell count. The recombinant virus was used to infect the DPSCs cells via multiplicity of infection (MOI) and induce the DPSCs reprogramming for IPCs. Immunofluorescence staining was used to measure the expression of proinsulin, FOXA2 and PDX1. ELISA method was used to detect the insulin secretion. The data was analyzed Using SPSS13.0 software package. RESULTS: DPSCs were isolated and cultured successfully. Cell surface highly expressed STRO-1 (98.01%), CDl46 (98.51%), CD34 (99.54%) and CD45 (24.08%). The multi-lineage differentiation capacity into osteoblasts, chondrocytes, and adipose was achieved. The recombinant hFOXA2 and hPDX1 lentivirus vectors were successfully constructed. Double enzyme digestion and sequencing appraisal showed that the sequence was fully consistent with GenBank retrieval. Virus packing efficiency was (96.15±0.17) % and (95.49±0.21) % respectively, and the infection titer was about 1.80±108 GTU/mL. The best MOI of the virus was 20. After inducing the cells to express proinsulin, FOXA2 and PDX1, insulin secretion volume was about 1.92 µmol/L. Compared with the uninduced group and control group, insulin secretion increased significantly (P<0.01). CONCLUSIONS: The recombinant transcription factor virus can activate cell reprogramming mechanism, form insulin-producing cells, and can be used for gene therapy of diabetes seed cells. Supported by Science and Technology Research Program of Shaanxi Province (2009K17-06) and Science and Technology Innovation as a Whole Plan Resources Leading Industry Key Technology (Chain) Project of Shaanxi Province (2011KTCL03-24).


Assuntos
Polpa Dentária , Vetores Genéticos , Lentivirus , Diferenciação Celular , Células Cultivadas , Fator 3-beta Nuclear de Hepatócito , Proteínas de Homeodomínio , Humanos , Insulina , Células-Tronco Mesenquimais , Osteoblastos , Células-Tronco , Dente Decíduo , Transativadores , Transfecção
14.
Zhonghua Nan Ke Xue ; 18(7): 661-4, 2012 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-22994054

RESUMO

OBJECTIVE: To evaluate the clinical effects of Compound Xuanju Capsule combined with tamoxifen citrate on the seminal parameters of men with idiopathic oligozoospermia. METHODS: We equally assigned 120 men with idiopathic oligozoospermia to receive Compound Xuanju Capsule plus tamoxifen citrate (experiment group) or tamoxifen citrate alone (control group). After 3 months of medication, we compared the concentration and total number of sperm with the baseline, and analyzed the influence of the duration of natural infertility on the therapeutic effect. RESULTS: Both the concentration and total number of sperm were significantly increased in both the experiment and the control groups after 3 months of medication as compared with the baseline (P < 0.05), and the increases were even more significant in the former than in the latter group (P < 0.05). The therapeutic efficacy was remarkably better in the patients with natural infertility < or = years than in those > 3 years (P < 0.05). CONCLUSION: Compound Xuanju Capsule combined with tamoxifen citrate produces satisfactory results in the treatment of idiopathic oligozoospermia, and therefore can be used as a first-line therapy for this disease.


Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Oligospermia/tratamento farmacológico , Tamoxifeno/uso terapêutico , Adulto , Quimioterapia Combinada , Humanos , Masculino , Fitoterapia , Resultado do Tratamento
15.
Zhongguo Zhong Yao Za Zhi ; 37(20): 3117-21, 2012 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-23311165

RESUMO

OBJECTIVE: To observe the effect of ginsenoside Rg1 on behavior and hippocampal amino acids in depressive-like rats. METHOD: SD rats were randomly divided into 5 groups: control, model, fluxetine, low dose ginsenoside Rg1 and high dose of ginsenoside Rg1. The chronic unpredictable mild stress (CUMS) was performed to induce depressive-like animal model. Fluxetine group was orally given fluxetine in dose of 10 mg x kg(-1) for 21 days, low dose ginsenoside Rg1 group was orally given ginsenoside Rg1 in dose of 20 mg x kg(-1) for 21 days, high dose ginsenoside Rg1 group was orally given ginsenoside Rg1 in dose of 40 mg x kg(-1) for 21 days. The control and model group was orally given saline for 21 days. The sucrose consumption was detected before and after the CUMS procedure. The horizontal and vertical activities of rats were determined by open-field test. HPLC was adopted to detect the contents of amino acids in hippocampus. RESULT: The sucrose consumption, horizontal and vertical activities in CUMS rats were decreased compared with those in control group. Compared with control group, the contents of glutamate (Glu) and aspartate (Asp) in hippocampus of CUMS group were increased, while the gamma amino butyric acid (GABA) and taurine (Tau) were decreased. Ginsenoside Rg1 treatment significantly increased the CUMS-induced decrease in sucrose consumption, horizontal and vertical activities. Administrated with ginsenoside Rg1 also decreased Glu and Asp and increased the GABA and Tau in hippocampus in a dose dependent manner. CONCLUSION: Ginsenoside Rg1 could alleviate the behavior changes of depressive-like rats, which might be related to regulate the levels of amino acids in hippocampus during CUMS and prevent the neuro-toxicity of excitatory amino acids.


Assuntos
Aminoácidos/metabolismo , Depressão/tratamento farmacológico , Depressão/psicologia , Ginsenosídeos/administração & dosagem , Hipocampo/metabolismo , Animais , Comportamento Animal/efeitos dos fármacos , Depressão/metabolismo , Modelos Animais de Doenças , Hipocampo/efeitos dos fármacos , Humanos , Masculino , Ratos , Ratos Sprague-Dawley
16.
Shanghai Kou Qiang Yi Xue ; 20(5): 454-8, 2011 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-22109358

RESUMO

PURPOSE: To clone the peptidylarginine deiminase(PAD) gene of Porphyromonas gingivalis(P.gingivalis) and to be expressed in E. coli under the best conditions. METHODS: With P.gingivalis strains ATCC33277 genomic DNA as a template, PCR was applied to obtain gene PAD which was then inserted into linear cloning vector PMD-18-T to construct clone recon. Recombinant PMD18-T-PAD was cloned and analyzed with PCR and restriction endonuclease,and PET-28a expression vector was digested by Xhol and Ncol,their products were linked to construct expression plasmid PET-28a-PAD under certain connection system. The recombinant expression plasmid PET-28a-PAD which had been confirmed correctly was transformed to E. coli competent cells BL21 and induce the expression of PAD with IPTG of different density and time. With His Tag monoclonal antibody as the first antibody, the expressed fusion protein was characterized by Western blot. RESULTS: DNA sequencing showed that the fragment was same as the sequence published in NCBI. Under the condition of 37 degrees centigrade, 0.5mmol/L IPTG, 250r/min shaking for 6 hours, the PAD could be highly expressed. CONCLUSIONS: The PAD is successfully cloned and expressed in E. coli which can be further uesd to study the immunity of PAD and the homologues antibody preparation.


Assuntos
Clonagem Molecular , Porphyromonas gingivalis , Células Cultivadas , Escherichia coli , Vetores Genéticos , Hidrolases , Plasmídeos , Reação em Cadeia da Polimerase , Desiminases de Arginina em Proteínas
17.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 29(2): 199-202, 2011 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-21598499

RESUMO

OBJECTIVE: To clone the glyceraldehydes 3-phosphate dehydrogenase (GAPDH) gene of Porphyromonas gingivalis (P. gingivalis) and to induce its fusion expression in E. coli. METHODS: GAPDH was obtained by PCR and was inserted into cloning vector pMD-18-T to construct clone recon. Double enzymes digest the recon pMD18-T-GAPDH and the prokaryotic expression vector pET-32a and then connect to get the expressing recon pET-32a-GAPDH. The recombinant expression plasmid which had been confirmed by enzymes digestion was transformed to E. coli competent cells BL21 and induced the expression of GAPDH with isopropyl beta-D-1-thiogalactopyranoside (IPTG) of different density. RESULTS: DNA sequencing showed that the fragment was 99.802% the same to the sequence published in NCBI. Under the best density, IPTG could be highly expressed. CONCLUSION: The GAPDH had been successfully cloned and expressed in E. coli which gets ready for the following experiment to study the immunity of GAPDH and the homologues antibody preparation.


Assuntos
Escherichia coli , Porphyromonas gingivalis , Células Cultivadas , Clonagem Molecular , Clonagem de Organismos , Vetores Genéticos , Gliceraldeído , Oxirredutases , Fosfatos , Reação em Cadeia da Polimerase
18.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 28(3): 241-5, 2010 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-20635648

RESUMO

OBJECTIVE: To clone the FimA gene of fimbriae from Porphyromonas gingivalis (P. gingivalis) and to construct prokaryotic expression vector which was induced in E.coli BL21(DE3)pLyS in the form of fusion protein expression and to identify, purify the product of its expression. METHODS: To clone the FimA gene of fimbriae from P. gingivalis and to construct prokaryotic expression vector pET15b-FimA vector which was transformed into the competent cells of BL21(DE3)pLyS. The expression of fusion protein was induced by isopropyl beta-D-1-thiogalactopyranoside (IPTG). With anti-6xHis Tag monoclonal antibody as the first antibody, the expressed fusion protein was characterized by Western blot and purified by Co(2+)-NTA affinity chromatography. RESULTS: Cloned FimA gene sequences and inserted into expression vector of the FimA sequences were related to the sequence in GenBank database showed 100% homology. IPTG induced and then identified by Western blot showed a fragment of 4.1 x 10(4) has been expressed. Co(2+)-NTA affinity chromatography column was used to obtain high concentrations of FimA purified protein. CONCLUSION: The recombinant prokaryotic expression vector of pET15b-FimA was constructed and was expressed and purified successfully in E. coli BL21 (DE3)pLyS. This study laid the experimental foundation to further prepare for monoclonal antibodies of fimbriae of P. gingivalis and to develop the subunit protein vaccine of prevention of periodontitis.


Assuntos
Escherichia coli , Porphyromonas gingivalis , Clonagem Molecular , Proteínas Recombinantes de Fusão , Proteínas Recombinantes
19.
Zhonghua Nan Ke Xue ; 16(10): 950-3, 2010 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-21243761

RESUMO

OBJECTIVE: To investigate the effect of Jinleng undershorts on the elevated scrotal temperature induced by varicocele as well as on other clinical symptoms of the disease. METHODS: Fifty-one varicocele patients received the treatment of wearing Jinleng undershorts for 30 min twice a day for a course of 90 days. Comparisons were made between the scrotal temperatures and other clinical symptoms of varicocele before and after the treatment. RESULTS: After 90 days of treatment with Jinleng undershorts, the left scrotal temperature of the varicocele patients was significantly reduced from (32.16 +/- 0.79) degrees C to (31.53 +/- 0.77) degrees C (P < 0.01), and the right scrotal temperature decreased from (31.91 +/- 0.73) degrees C to (31.81 +/- 0.63) degrees C (P > 0.05). Compared with pretreatment, significant improvement was found in such symptoms as wetness, fever and swelling of the scrotum, backache, headache, dizziness, fatigue and anxiety (P < 0.05), as well as in testicular pain (P < 0.01) and IIEF-5 score, which was increased from 15.89 +/- 6.13 to 20.04 +/- 3.87 (P < 0.01). CONCLUSION: Jinleng undershorts can be used for the treatment of mild and moderate varicocele.


Assuntos
Vestuário , Medicina Tradicional Chinesa , Varicocele/terapia , Temperatura Corporal , Humanos , Masculino , Escroto
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