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1.
Chinese Journal of Preventive Medicine ; (12): 386-392, 2023.
Artigo em Chinês | WPRIM | ID: wpr-969900

RESUMO

Objective: To characterize the prevalence and genomic epidemiology of Vibrio parahaemolyticus from acute diarrheal patients in Shenzhen City from 2013 to 2021. Methods: Based on the Shenzhen Infectious Diarrhea Surveillance System, acute diarrheal patients were actively monitored in sentinel hospitals from 2013 to 2021. Whole-genome sequencing (WGS) of Vibrio parahaemolyticus isolates was performed, and the genomic population structure, serotypes, virulence genes and multilocus sequence typing were analyzed. Outbreak clusters from 2019 to 2021 were explored based on single-nucleotide polymorphism analysis. Results: A total of 48 623 acute diarrhea cases were monitored in 15 sentinel hospitals from 2013 to 2021, and 1 135 Vibrio parahaemolyticus strains were isolated, with a positive isolation rate of 2.3%. Qualified whole-genome sequencing data of 852 isolates were obtained. Eighty-nine serotypes, 21 known ST types and 5 new ST types were identified by sequence analysis, and 93.2% of strains were detected with toxin profile of tdh+trh-. 8 clonal groups (CGs) were captured, with CG3 as the absolute predominance, followed by CG189. The CG3 group was dominated by O3:K6 serotype and ST3 sequence type, while CG189 group was mainly O4:KUT, O4:K8 serotypes and ST189a and ST189 type. A total of 13 clusters were identified, containing 154 cases. About 30 outbreak clusters with 29 outbreak clusters caused by CG3 strains from 2019 to 2021. Conclusion: Vibrio parahaemolyticus is a major pathogen of acute infectious diarrhea in Shenzhen City, with diverse population structures. CG3 and CG189 have been prevalent and predominant in Shenzhen City for a long time. Scattered outbreaks and persistent sources of contamination ignored by traditional methods could be captured by WGS analysis. Tracing the source of epidemic clone groups and taking precise prevention and control measures are expected to significantly reduce the burden of diarrhea diseases caused by Vibrio parahaemolyticus infection in Shenzhen City.


Assuntos
Humanos , Vibrio parahaemolyticus/genética , Diarreia/epidemiologia , Doenças Transmitidas por Alimentos/epidemiologia , Sorogrupo , Genômica , Disenteria , Vibrioses/epidemiologia , Sorotipagem
2.
Acta Pharmaceutica Sinica ; (12): 1954-1961, 2022.
Artigo em Chinês | WPRIM | ID: wpr-936582

RESUMO

With the acceleration of the aging process of our country's population, the impact of aging-related diseases - Alzheimer's disease (AD) on society and families has become increasingly prominent. AD is caused by multiple mechanisms, and the pathogenesis has not been fully elucidated. Most of the clinical treatments are single therapy, which mainly focuses on improving symptoms and are difficult to reverse the disease process. Therefore, the development of drugs that can both improve symptoms and reverse the disease process is extremely urgent in clinical. Increasing number of studies has shown that traditional Chinese medicine plays an important role in the prevention and treatment of AD. The natural products have many advantages, such as novel structures, multiple targets and diverse activities, which can be used as an important source of leading compounds for the treatment of AD. The review summarizes the main clinical treatment methods and the research progress of natural ingredients in traditional Chinese medicine, and provides a reference for the follow-up clinical treatment of AD combined with the advantages of traditional Chinese medicine treatment.

3.
China Journal of Orthopaedics and Traumatology ; (12): 1017-1022, 2020.
Artigo em Chinês | WPRIM | ID: wpr-879344

RESUMO

OBJECTIVE@#To analyze the clinical efficacy of hip arthroplasty with femoral calcar prosthesis and proximal femoral nail fixation(PFNA) in the treatment of elderly patients(≥80 years old) with unstable intertrochanteric fractures(Evans Ⅲ, Ⅳ).@*METHODS@#From June 2016 to March 2018, 60 elderly patients with unstable intertrochanteric fractures treated with prosthetic replacement and PFNA were retrospectively analyzed. According to the surgical methods, they were divided into PFNA group and prosthesis group. In PFNA group there were including 21 males and 15 females, with an average age of(84.3± 2.9) years old;in the prosthetic group, there were 10 males and 14 females with an average age of (82.9±2.4) years old. The operation time, hemoglobin difference between preoperative and postoperative 1 day, postoperative ambulation time, hospitalization time and complications were observed and compared between the two groups. Harris hip score was performed 3 and 12 months after operation.@*RESULTS@#All patients were followed up for 12 to 24 months (19.3±4.8) months. One patient in the prosthesis group died of lung cancer one year later and the follow-up was terminated. The operation time of prosthetic group was longer than that of PFNA group(@*CONCLUSION@#The elderly patients with intertrochanteric arthroplasty can reduce the burden of intertrochanteric arthroplasty and improve the quality of life.


Assuntos
Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pinos Ortopédicos , Fixação Intramedular de Fraturas , Fraturas do Quadril/cirurgia , Pacientes , Qualidade de Vida , Estudos Retrospectivos , Resultado do Tratamento
4.
Neuroscience Bulletin ; (6): 261-269, 2018.
Artigo em Inglês | WPRIM | ID: wpr-777069

RESUMO

Hyperphosphorylated tau is the major protein component of neurofibrillary tangles in the brains of patients with Alzheimer's disease (AD). However, the mechanism underlying tau hyperphosphorylation is not fully understood. Here, we demonstrated that exogenously expressed wild-type human tau40 was detectable in the phosphorylated form at multiple AD-associated sites in cytoplasmic and nuclear fractions from HEK293 cells. Among these sites, tau phosphorylated at Thr205 and Ser214 was almost exclusively found in the nuclear fraction at the conditions used in the present study. With the intracellular tau accumulation, the Ca concentration was significantly increased in both cytoplasmic and nuclear fractions. Further studies using site-specific mutagenesis and pharmacological treatment demonstrated that phosphorylation of tau at Thr205 increased nuclear Ca concentration with a simultaneous increase in the phosphorylation of Ca/calmodulin-dependent protein kinase IV (CaMKIV) at Ser196. On the other hand, phosphorylation of tau at Ser214 did not significantly change the nuclear Ca/CaMKIV signaling. Finally, expressing calmodulin-binding protein-4 that disrupts formation of the Ca/calmodulin complex abolished the okadaic acid-induced tau hyperphosphorylation in the nuclear fraction. We conclude that the intracellular accumulation of phosphorylated tau, as detected in the brains of AD patients, can trigger nuclear Ca/CaMKIV signaling, which in turn aggravates tau hyperphosphorylation. Our findings provide new insights for tauopathies: hyperphosphorylation of intracellular tau and an increased Ca concentration may induce a self-perpetuating harmful loop to promote neurodegeneration.


Assuntos
Humanos , Doença de Alzheimer , Metabolismo , Patologia , Cálcio , Metabolismo , Proteína Quinase Tipo 4 Dependente de Cálcio-Calmodulina , Metabolismo , Núcleo Celular , Metabolismo , Ativação Enzimática , Fisiologia , Células HEK293 , Neurônios , Metabolismo , Patologia , Fosforilação , Transdução de Sinais , Fisiologia , Proteínas tau , Metabolismo
5.
Chinese Journal of Immunology ; (12): 325-330, 2018.
Artigo em Chinês | WPRIM | ID: wpr-702727

RESUMO

Objective:To study the expression and interaction between miR-194 and PTPN12 in the process of age-related atrophy of thymus for clarifying the regulatory mechanism in the process of this disease.Methods:C57BL/6 mouse were divided into 4 groups as 1 month,6 months,10 months and 19 months old and each group has 6 cases.Thymus tissue was removed and thymic stromal cells were isolated.And thymus epithelial cells were washed out by CD45 antibody and LS column after anesthesia.Fluorescence quantitative real-time PCR and Western blot were used to detect the changes of miR-194 and PTPN12 gene expression in thymus epithelial cells with aging.miR-194 and PTPN12 luciferase reporter vectors were transfected into HEK293 cells,and the auto fluorescence values were detected at 24 h and 48 h,respectively in vitro.Results:The expression level of miR-194 decreased (P<0.05),while the expression level of PTPN12 mRNA increased (P<0.05) as the age increased.And the correlation between miR-194 and PTPN12 mRNA expression was found to be negative(P<0.05).In vitro,luciferase reporter gene results show that miR-194 has a direct effect on the 3'UTR region of PTPN12 gene and had the highest binding efficiency in 48 h.Conclusion:PTPN12 is one of the target genes of miR-194,which is involved in the aging process of thymus and is an important factor regulating the function of thymic ep-ithelial cells.

6.
Journal of Southern Medical University ; (12): 268-273, 2018.
Artigo em Chinês | WPRIM | ID: wpr-690477

RESUMO

<p><b>OBJECTIVE</b>To investigate the role of p38MAPK signaling pathway in autophagy of intestinal epithelial cells induced by spvB of S.typhimurium.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>The inhibitory effect of spvB gene on autophagy in intestinal epithelial cells is related with the negative regulation of p38MAPK signaling pathway.</p>

7.
Chinese Journal of Contemporary Pediatrics ; (12): 1278-1284, 2017.
Artigo em Chinês | WPRIM | ID: wpr-300407

RESUMO

<p><b>OBJECTIVE</b>To investigate the molecular mechanism of action of BET bromodomain inhibitor JQ1 in treating airway remodeling in asthmatic mice.</p><p><b>METHODS</b>A total of 24 mice were randomly divided into control group, ovalbumin (OVA)-induced asthma group (OVA group), and JQ1 intervention group (JQ1+OVA group), with 8 mice in each group. OVA sensitization/challenge was performed to establish a mouse model of asthma. At 1 hour before challenge, the mice in the JQ1+OVA group were given intraperitoneal injection of JQ1 solution (50 μg/g). Bronchoalveolar lavage fluid (BALF) and lung tissue samples were collected at 24 hours after the last challenge, and the total number of cells and percentage of eosinophils in BALF were calculated. Pathological staining was performed to observe histopathological changes in lung tissue. RT-PCR and Western blot were used to measure the mRNA and protein expression of E-cadherin and vimentin during epithelial-mesenchymal transition (EMT).</p><p><b>RESULTS</b>Compared with the control group, the OVA group had marked infiltration of inflammatory cells in the airway, thickening of the airway wall, increased secretion of mucus, and increases in the total number of cells and percentage of eosinophils in BALF (P<0.01). Compared with the OVA group, the JQ1+OVA group had significantly alleviated airway inflammatory response and significant reductions in the total number of cells and percentage of eosinophils in BALF (P<0.01). Compared with the control group, the OVA group had significant reductions in the mRNA and protein expression of E-cadherin and significant increases in the mRNA and protein expression of vimentin (P<0.01); compared with the OVA group, the JQ1+OVA group had significant increases in the mRNA and protein expression of E-cadherin and significant reductions in the mRNA and protein expression of vimentin (P<0.01); there were no significant differences in these indices between the JQ1+OVA group and the control group (P>0.05).</p><p><b>CONCLUSIONS</b>Mice with OVA-induced asthma have airway remodeling during EMT. BET bromodomain inhibitor JQ1 can reduce airway inflammation, inhibit EMT, and alleviate airway remodeling, which provides a new direction for the treatment of asthma.</p>


Assuntos
Animais , Feminino , Camundongos , Remodelação das Vias Aéreas , Asma , Tratamento Farmacológico , Azepinas , Farmacologia , Caderinas , Genética , Transição Epitelial-Mesenquimal , Proteínas Nucleares , Ovalbumina , Alergia e Imunologia , RNA Mensageiro , Fatores de Transcrição , Triazóis , Farmacologia , Vimentina , Genética
8.
Chinese Journal of Epidemiology ; (12): 609-613, 2013.
Artigo em Chinês | WPRIM | ID: wpr-318340

RESUMO

<p><b>OBJECTIVE</b>To determine the occurrence and distribution of specific clones of pathogenic Vibrio parahaemolyticus(VP)isolated in Shenzhen and to assess the relationship between serotype O3:K6 and the globally distributed pandemic clone.</p><p><b>METHODS</b>A total of 1005 VPs isolated from diarrhea patients in 2002-2008 were sero-typed. Real-time PCR was used to detect the virulence genes tlh, toxR, tdh, trh and orf8 in 281 isolates from 68 different serotypes. The main serotypes were typed by pulsed field gel electrophoresis(PFGE). Strains with dominant serotypes and PFGE patterns were assayed by GS-PCR and toxRS sequencing for the identification of pandemic clone. Multilocus sequence typing(MLST)analysis was reserved for exemplary 41 O3 : K6 and O1 : K25 isolates.</p><p><b>RESULTS</b>Seventy-nine serotypes were observed among the 1005 isolates, including O3 : K6(57.9%), O4 : K8(8.16%), O1 : KUT(5.87%), O1 : K25(5.27%), O4 : K68(1.39%), O1 : K56(1.39%) and O9 : K44(0.99%). Most of the strains(99.36%)showed PCR positive to tlh, toxR, and tdh but eleven strains were tdh negative. MLST showed that all the 36 O3 : K6 isolates belonged to ST3 and all the 5 O4 : K8 strains were ST189. These results matched the description of the pandemic VP clone.</p><p><b>CONCLUSION</b>A recognizable burden of diarrheal illness caused by VP had been seen in Shenzhen. Results from serotyping indicated that although there existing a large variety of diversities, the dominant serotype appeared to be O3 : K6. VP isolates identified in Shenzhen mainly showed as tdh positive but trh negative, in consistent with the current pandemic O3 : K6 clone. The pandemic O3 : K6 clone did appear to co-exist with other clones of O3 : K6, as well as O4 : K8,O1 : K25. Potential outbreak of VP could be monitored through the laboratory-based surveillance programs, suggesting that the strategies related to prevention and control of VP should be prioritized in Shenzhen.</p>


Assuntos
Humanos , China , Epidemiologia , Eletroforese em Gel de Campo Pulsado , Tipagem de Sequências Multilocus , Reação em Cadeia da Polimerase em Tempo Real , Sorotipagem , Vibrioses , Epidemiologia , Microbiologia , Vibrio parahaemolyticus , Genética , Virulência
9.
Chinese Traditional and Herbal Drugs ; (24): 3496-3502, 2013.
Artigo em Chinês | WPRIM | ID: wpr-854992

RESUMO

Objective: To investigate the intervention of quercetin on rats with uric acid nephropathy (UAN) and its molecular mechanisms. Methods: Rats were divided into six groups including normal, model, quercetin (25, 50, and 100 mg/kg)-treated, and allopurinol (5 mg/kg)-treated groups. Adenine (100 mg/kg) and ethambutol (250 mg/kg) were ig given to rats once daily for consecutive three weeks to establish UAN model. Quercetin (25, 50, and 100 mg/kg) and allopurinol (5 mg/kg) were initially ig given to UAN rats 1 h after adenine and ethambutol had been given. Rat renal histopathological changes were observed; The levels of uric acid (Sur and Uur), creatinine (Scr and Ucr), and blood urea nitrogen (BUN) in serum and urine were detected; The fractional excretion of uric acid (FEUA) was calculated. Simultaneously, mRNA and protein levels of components of NOD like receptor protein 3 (NLRP3) inflammasomes (NLRP3, ASC, and Caspase-1) and key factors in Toll like receptors (TLRs) signaling pathways (TLR2 and TLR4) were analyzed by RT-PCR and Western blotting methods, respectively. Results: Compared with the model group, quecetin and allopurinol reduced Sur and Scr levels significantly, increased Uur and Ucr excretion in 24 h, and recovered FEUA in UAN rats. Moreover, the mRNA and protein expression of NLRP3, ASC, Caspase-1, TLR2, and TLR4 was up-regulated in UAN rat kidneys, which could be reversed by the treatment of quercetin and allopurinol. Conclusion: These findings suggest that the activated NLRP3 inflammasomes and TLRs signaling pathways could play the causal roles in UAN pathogenesis. Quercetin could ameliorate UAN by regulating NLRP3 inflammasomes and TLRs signaling pathways.

10.
Acta Physiologica Sinica ; (6): 289-295, 2012.
Artigo em Chinês | WPRIM | ID: wpr-335911

RESUMO

To investigate the effect of Ca(2+)-sensing receptor (CaR) on Spermine-induced extracellular Ca(2+) influx and NO generation in human umbilical vein endothelial cells (HUVEC), the small interference RNA (siRNA) specifically targeting CaR gene was designed, synthesized and transfected into HUVEC according to the cDNA sequence of human CaR gene in GenBank. The transfection efficiency and the interference efficiency of CaR protein were determined by laser scanning confocal microscopy and Western blot, respectively. Intracellular Ca(2+) concentration ([Ca(2+)](i)) was measured by Fura-2/AM loading. The production of NO and the activity of endothelial nitric oxide synthase (eNOS) were determined by the DAF-FM diacetate (DAF-FM DA). Western blot results demonstrated that siRNA targeting the CaR specifically decreased the expression of CaR protein in CaR siRNA group 48 h after transfection (P < 0.05). At the same time, the Spermine-induced [Ca(2+)](i), eNOS activity and NO generation were also significantly reduced (P < 0.05) in CaR siRNA group compared with those in the untransfected or negative siRNA transfected group. In conclusion, the present study suggests that the CaR plays an important role in the Spermine-evoked process of extracellular Ca(2+) influx and NO generation in HUVEC.


Assuntos
Humanos , Cálcio , Fisiologia , Células Cultivadas , Células Endoteliais da Veia Umbilical Humana , Fisiologia , Óxido Nítrico , Fisiologia , Óxido Nítrico Sintase Tipo III , Fisiologia , RNA Interferente Pequeno , Receptores de Detecção de Cálcio , Genética , Fisiologia , Espermina , Farmacologia , Transfecção
11.
Chinese Journal of Applied Physiology ; (6): 289-293, 2012.
Artigo em Chinês | WPRIM | ID: wpr-329884

RESUMO

<p><b>OBJECTIVE</b>To investigate whether homocysteine (Hcy) participates the proliferation of the spontaneously hypertensive rat(SHR) vascular smooth muscle cell (VSMCs) and the molecular mechanism.</p><p><b>METHODS</b>The rat's arota were removed. The primary SHR VSMCs were isolated and cultured in vitro, then the SHR VSMCs were divided into four groups: (1) control group, (2) Hcy group, (3) 18alpha-glycyrrhetinic acid (GA) group, (4) Hcy + 18alpha-GA group. We detected proliferation of the SHR VSMCs by MTT and flow cytometry. The expression and co-localization of the connexin (Cx) 43 and Cx40 proteins in the SHR VSMCs were deteced by immunofluorescence. The expression of the Cx43 and Cx40 proteins in SHR VSMCs were detected by Western blot. The molecular dye transfer method (scrape dye transfer method) was applied to detect the gap junction function in the SHR VSMCs.</p><p><b>RESULTS</b>(1) The Cx43 and Cx40 proteins expression in the SHR VSMCs were positive, confocal microscopy supported the co-localization of Cx43 and Cx40 in the cytoplasm. (2) The S value deteced by cell cycle and A value detected by MTT in the Hcy group were increased obviously compared with those in the control group (P < 0.05), decreased in 18alpha-GA group (P < 0.05). Compared with the Hcy group, the S and A value in the Hcy + 18alpha-GA group were significantly decreased, respectively (P < 0.05). (3) The expression of Cx43 and Cx40 proteins in Hcy group were increased compared with the control group (P < 0.05), decreased in 18alpha-GA group (P < 0.05). Compared with the Hcy group, the expression of Cx43 and Cx40 proteins in the Hcy + 18alpha-GA group were significantly decreased, respectively (P < 0.05). (4) The function of gap junction detected by scrape dye transfer method in the Hcy group were enhanced compared with the control group (P < 0.05), weakened in the 18alpha-GA group (P < 0.05). Compared with the Hcy group,the function of gap junction in the Hcy + 18alpha-GA group was significantly weakened (P < 0.05).</p><p><b>CONCLUSION</b>Hcy can enhance the function of gap junctional to stimulate the proliferation of SHR VSMCs through the expression of Cx43 and Cx40 proteins promoted.</p>


Assuntos
Animais , Ratos , Proliferação de Células , Células Cultivadas , Conexina 43 , Metabolismo , Conexinas , Metabolismo , Junções Comunicantes , Metabolismo , Ácido Glicirretínico , Farmacologia , Homocisteína , Farmacologia , Músculo Liso Vascular , Biologia Celular , Metabolismo , Miócitos de Músculo Liso , Biologia Celular , Metabolismo , Ratos Endogâmicos SHR
12.
Chinese Journal of Preventive Medicine ; (12): 899-903, 2011.
Artigo em Chinês | WPRIM | ID: wpr-266076

RESUMO

<p><b>OBJECTIVE</b>To study the characteristics of the strains of Salmonella enterica (S. enterica) serovar Senftenberg lacking Salmonella pathogenicity island 1 (SPI-1).</p><p><b>METHODS</b>A total of 10 strains of S. enterica serovar Senftenberg were isolated from 10 cases of diarrhea patients. Pulsed field gel electrophoresis (PFGE), PCR, sequencing techniques and cell invasion test were adapted to study the molecular types and invasiveness of the genes and cells; and made a comparison between the 10 strains and the strains (C02013) isolated in Shenzhen in 2002.</p><p><b>RESULTS</b>The 10 Senftenberg isolated (S09007-S09012, S09014-S09017) in Shanghai showed three PFGE patterns, which were significantly different from the strains isolated in Shenzhen. PCR-amplified results indicated the invasion gene (invA), secreted effector protein gene (sipA) and gene fragments as fhlA-hilA, hilA-spaP and spaP-invH in the 10 strains of SPI-1 were all negative. The sequencing results revealed that the 10 strains isolated in Shanghai lacked most parts of SPI-1 genes, as fragments from orgA to invH and parts of orgA gene itself; however, compared with strains isolated in Shenzhen, the sprB-orgC gene existed. The missing parts of genes were replaced by a simple insertion sequence (IS) of 1000 bp in the strains isolated both in Shenzhen in 2002 and in Shanghai in 2006. The invasiveness rates of the 10 strains (S09007-S09012, S09014-S09017) towards Hela cells were (0.0053 ± 0.0024)%, (0.0046 ± 0.0006)%, (0.0047 ± 0.0003)%, (0.0064 ± 0.0012)%, (0.0065 ± 0.0011)%, (0.0070 ± 0.0020)%, (0.0115 ± 0.0030)%, (0.0099 ± 0.0039)%, (0.0180 ± 0.0135)% and (0.0031 ± 0.0012)%, respectively; which were all significantly lower than the rate of invA-positive control strain STM1344 ((5.0800 ± 0.6333)%); lower or close to the rate of invA-lacked artificial-mutated strain STMinvA-((0.0193 ± 0.0045)%).</p><p><b>CONCLUSION</b>SPI-1 genes are not essential for the diarrhea caused by S. enterica serovar Senftenberg.</p>


Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Técnicas de Tipagem Bacteriana , Diarreia , Microbiologia , Fezes , Microbiologia , Genes Bacterianos , Ilhas Genômicas , Células HeLa , Salmonella enterica , Genética , Virulência
13.
Acta Physiologica Sinica ; (6): 39-47, 2011.
Artigo em Chinês | WPRIM | ID: wpr-337706

RESUMO

Although the function of extracellular Ca(2+)-sensing receptor (CaR) is known, the regulatory mechanism of the CaR function remains to be clarified. The purpose of the present study was to investigate the effect of caveolin-1 (Cav-1) on CaR-induced extracellular Ca(2+) influx by using acute caveolae disruption with Filipin or siRNA targeted to the Cav-1 in human umbilical vein endothelial cells (HUVECs). Intracellular Ca(2+) concentration ([Ca(2+)](i)) was detected by Fura-2/AM loading. The results showed that different concentrations of extracellular Ca(2+) failed to increase [Ca(2+)](i), while the CaR agonist Spermine (2 mmol/L) resulted in an increase in [Ca(2+)](i) that was diminished in buffer without Ca(2+) (P<0.05). No matter in buffer with or without 2 mmol/L Ca(2+), the [Ca(2+)](i) increase induced by Spermine in HUVECs was abolished after inhibition of CaR by a negative allosteric modulator Calhex231 (1 μmol/L) (P<0.05), conversely, the effect of Spermine on the increase in [Ca(2+)](i) in HUVECs was further augmented after acute caveolae disruption with Filipin (1.5 μg/mL) or transfection with siRNA targeted to the Cav-1 (P<0.05). This indicated that Cav-1 produced an inhibition of CaR-induced extracellular Ca(2+) influx. As to the biological mechanism of Cav-1-induced inhibition, immunofluorescence technique showed that both CaR and Cav-1 were present in HUVECs, and confocal microscopy supported the co-localization of CaR and Cav-1 on the plasma membrane. Functionally, the Cav-1 protein expression was decreased in HUVECs transfected with siRNA targeted to the Cav-1 (P<0.05); simultaneously, the CaR membrane protein expression was decreased (P<0.05), whereas CaR total protein level was unaffected (P>0.05). In conclusion, the present study suggests that CaR and Cav-1 co-localize on the plasma membrane in HUVECs and CaR-induced Ca(2+) influx is down-regulated by binding with Cav-1, and the mechanism involves the effect of Cav-1 on CaR localization on the plasma membrane and attenuating the CaR response to the agonist.


Assuntos
Humanos , Cálcio , Metabolismo , Canais de Cálcio , Metabolismo , Caveolina 1 , Fisiologia , Células Cultivadas , Regulação para Baixo , Filipina , Farmacologia , Células Endoteliais da Veia Umbilical Humana , Biologia Celular , Metabolismo , Receptores de Detecção de Cálcio , Fisiologia , Espermina , Farmacologia
14.
Chinese Journal of Epidemiology ; (12): 51-55, 2010.
Artigo em Chinês | WPRIM | ID: wpr-321001

RESUMO

Objective To study the infection status and the molecular characteristics of Vibrio parahaemolyticus isolated from diarrheal patients in Shenzhen, in 2007 to 2008 and to provide evidence for the prevention and control of diarrheal diseases caused by Vibrio parahaemolyticus. Methods More than 80 fecal specimens from four sentinel surveillance hospitals were collected and cultured each month. A total of 361 isolates of Vibrio parahaemolyticus were sero-typed and examined by real-time PCR for the presence of two major virulence genes, tdh and trh. Of 361 strains, 60 O3: K6 strains isolated from six suspected outbreaks in August, 2007 and in September, 2008 were typed by pulsed-field gel electrophoresis (PFGE). Results 4384 stool samples were detected in four sentinel surveillance hospitals and with 361 Vibrio parahaemolyticus strains isolated that belonged to 28 serotypes. Serotype O3:K6, O4:K8 and O1:KUT accounted for 67.90%, 7.50% and 6.10%, respectively. Of 361 strains, 337 strains belonged to tdh + trh- , 11 strains were tdh-trh- and 13 strains were tdh + trh +. The most prevalent serotype which caused diarrheal diseases was tdh + trh-in Shenzhen. The 60 isolates were discriminated into twenty different PFGE patterns, which belonged to three clones. Among the 60 isolates, most of the PFGE patterns of isolates from the suspected outbreak locations were identical and some strains isolated from different year were different. Conclusion Vibrio parahaemolyticus isolates in Shenzhen were dominated by O3:K6 strains. Most of these isolates carried tdh gene and few carried trh gene. Meanwhile, the identical patterns of isolates from 6 suspected outbreaks locations demonstrated that Vibrio parahaemolyticus outbreaks occurred in July 2007 and in September 2008 in Shenzhen. However, the dominated strains' PFGE patterns were different each year, indicating that the sources of Vibrio parahaemolyticus had a multiplex nature and the multiplex sources such as water, sea food and pickled products should be integrated monitored. Laboratory based surveillance of diarrheal diseases could contribute in establishing early warning system for the better prevention and control of diarrheal diseases.

15.
Chinese Journal of Medical Genetics ; (6): 463-468, 2010.
Artigo em Chinês | WPRIM | ID: wpr-234381

RESUMO

<p><b>OBJECTIVE</b>To investigate the association of the transforming growth factor- beta 1 (TGF- beta 1) gene polymorphisms and blood TGF- beta 1 level with essential hypertension (EH) in Kazakh Chinese.</p><p><b>METHODS</b>The polymorphisms of TGF- beta 1 gene in 354 Kazakh EH patients and 435 healthy controls were detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing. The blood level of TGF- beta 1 was quantified using specific sandwich ELISA.</p><p><b>RESULTS</b>The frequencies of genotypes GG, GC and alleles G and C of +915G/C in Xinjiang Kazakh were 97.9%, 2.1% and 98.77%, 1.23%, respectively. No significantly difference was found between EH patients and controls (P>0.05). The frequencies of genotypes TT, TC, CC and alleles T and C of +869T/C in controls was 25.97%, 46.67%, 27.36%, 49.3% and 50.7%, respectively, the CC genotype or C allele in EH patients had significantly higher frequencies than controls [41.60% vs. 27.36%, and 62.2% vs. 50.7%, respectively (P<0.05)]. It was also shown that TGF- beta 1 +869 C allele carriers had significantly increased risk of EH compared with T allele carriers (OR=1.60, P=0.00). There was linkage disequilibrium (LD) between the two polymorphisms. The frequency of haplotype C-G in the EH group was significantly higher than that in controls (61.6% vs. 49.8%, P<0.05). There were no differences in TGF- beta 1 level among different genotypes or alleles in both +869T/C and +915G/C loci (P>0.05).</p><p><b>CONCLUSION</b>The frequency of +915G/C variation of the TGF- beta 1 gene was very low in Kazakh and there was no homozygous variation. The +869 C allele was likely the genetic susceptibility factor for EH in the population. There was linkage disequilibrium in the polymorphisms of +869T/C and +915G/C. Haplotype C-G was the risk factor of EH.</p>


Assuntos
Adulto , Humanos , Masculino , Pessoa de Meia-Idade , Alelos , Povo Asiático , Etnologia , Genética , Predisposição Genética para Doença , Genótipo , Hipertensão , Epidemiologia , Genética , Desequilíbrio de Ligação , Genética , Polimorfismo Genético , Fatores de Risco , Fator de Crescimento Transformador beta1 , Genética
16.
Acta Pharmaceutica Sinica ; (12): 1239-1246, 2010.
Artigo em Chinês | WPRIM | ID: wpr-354521

RESUMO

The effects of mangiferin on uric acid excretion, kidney function and related renal transporters were investigated in hyperuricemic mice induced by potassium oxonate. Mice were divided into normal control group, and 5 hyperuricemic groups with model control, 50, 100, and 200 mg x kg(-1) mangiferin, and 5 mg x kg(-1) allopurinol. Mice were administered by gavage once daily with 250 mg x kg(-1) potassium oxonate for seven consecutive days to create the model. And 3 doses of mangiferin were orally initiated on the day 1 h after potassium oxonate was given, separately. Serum uric acid, creatinine and urea nitrogon levels, as well as urinary uric acid creatinine levels were measured. Mouse uromodulin (mUMOD) levels in serum, urine and kidney were determined by ELISA method. The mRNA and protein levels of related renal transporters were assayed by RT-PCR and Western blotting methods, respectively. Compared to model group, mangiferin significantly reduced serum uric acid, creatinine and urea nitrogon levels, increased 24 h uric acid and creatinine excretion, and fractional excretion of uric acid in hyperuricemic mice, exhibiting uric acid excretion enhancement and kidney function improvement. Mangiferin was found to down-regulate mRNA and protein levels of urate transporter 1 (mURAT1) and glucose transporter 9 (mGLUT9), as well as up-regulate organic anion transporter 1 (mOAT1) in the kidney of hyperuricemic mice. These findings suggested that mangiferin might enhance uric acid excretion and in turn reduce serum uric acid level through the decrease of uric acid reabsorption and the increase of uric acid secretion in hyperuricemic mice. Moreover, mangiferin remarkably up-regulated expression levels of renal organic cation and carnitine transporters (mOCT1, mOCT2, mOCTN1 and mOCTN2), increased urine mUMOD levels, as well as decreased serum and kidney mUMOD levels in hyperuricemic mice, which might be involved in mangiferin-mediated renal protective action.


Assuntos
Animais , Masculino , Camundongos , Nitrogênio da Ureia Sanguínea , Proteínas de Transporte , Genética , Metabolismo , Creatinina , Sangue , Proteínas Facilitadoras de Transporte de Glucose , Genética , Metabolismo , Hiperuricemia , Sangue , Urina , Rim , Metabolismo , Proteínas de Membrana , Genética , Metabolismo , Fator 1 de Transcrição de Octâmero , Genética , Metabolismo , Proteína 1 Transportadora de Ânions Orgânicos , Genética , Metabolismo , Transportadores de Ânions Orgânicos , Genética , Metabolismo , Proteínas de Transporte de Cátions Orgânicos , Genética , Metabolismo , Transportador 2 de Cátion Orgânico , Ácido Oxônico , Substâncias Protetoras , Farmacologia , RNA Mensageiro , Metabolismo , Distribuição Aleatória , Membro 5 da Família 22 de Carreadores de Soluto , Ácido Úrico , Sangue , Urina , Uromodulina , Sangue , Urina , Xantonas , Farmacologia
17.
Chinese Journal of Epidemiology ; (12): 933-937, 2009.
Artigo em Chinês | WPRIM | ID: wpr-321094

RESUMO

ation were complicated, with the characteristics as the obvious decreasing number of patients, with no food-borne isolates in 2007.

18.
Chinese Journal of Preventive Medicine ; (12): 827-830, 2008.
Artigo em Chinês | WPRIM | ID: wpr-242708

RESUMO

<p><b>OBJECTIVE</b>To investigate the application of pulsed-field gel electrophoresis (PFGE) in food-borne outbreak.</p><p><b>METHODS</b>Pathogens were isolated and further characteristics identified by traditional methods. The strains isolated were carried out with molecular typing with using PFGE. PFGE was performed by Laboratory Directions for molecular subtyping of Salmonella by PFGE (CDC, USA) and the results of PFGE were analyzed by BioNumerics soft.</p><p><b>RESULTS</b>Totally 14 Salmonella serotype Muenchen strains were isolated from 19 patients, 3 of 9 suspicious foods were positive for S. muenchen and 7 strains were isolated from 18 cooks. The biochemistry characterization and antimicrobial susceptibility of all the strains isolated were the same. 23 S. muenchen isolates were all shown indistinguishable by PFGE.</p><p><b>CONCLUSION</b>PFGE should play a key role in identifying the outbreak-associated isolates and distinguishing them from unrelated sporadic isolates. It might also demonstrate that the genetic fingerprints of serotype Muenchen isolates derived from patients were indistinguishable from those derived from drinks. PFGE might provide precise information on bacterial food-borne pathogens, promptly identify the source of infection, and effectively prevent from spreading. It should be one of the early warning method on controlling outbreak of the food-borne disease.</p>


Assuntos
Humanos , China , Epidemiologia , Surtos de Doenças , Eletroforese em Gel de Campo Pulsado , Métodos , Testes de Sensibilidade Microbiana , Intoxicação Alimentar por Salmonella , Epidemiologia , Microbiologia , Salmonella enterica , Classificação , Sorotipagem
19.
Chinese Journal of Preventive Medicine ; (12): 317-320, 2008.
Artigo em Chinês | WPRIM | ID: wpr-352480

RESUMO

<p><b>OBJECTIVE</b>To analyze the genetic relations of Shigella isolated from Shenzhen in 2001-2006 and develop primary molecular subtyping surveillance network of Shigella.</p><p><b>METHODS</b>Chromosomal DNAs from 55 isolated in agarose were digested with the restriction enzyme Xba I, and then were analyzed by pulsed-field gel electrophoresis. Pulsed-field gel electrophoresis (PFGE) patterns were clustered using BioNumerics software.</p><p><b>RESULTS</b>All 41 distinctive PFGE patterns were identified among 55 strains. 32 strains belonged to one cluster. Differences were observed in other strains.</p><p><b>CONCLUSION</b>Both genetic-related clones and non-related clones of Shigella existed in Shenzhen. The development of PFGE molecular subtyping surveillance network would contribute to the active surveillance, outbreak investigation and source tracking for Shigellosis.</p>


Assuntos
Humanos , Técnicas de Tipagem Bacteriana , China , Eletroforese em Gel de Campo Pulsado , Métodos , Fezes , Microbiologia , Shigella , Classificação
20.
Acta Academiae Medicinae Sinicae ; (6): 474-478, 2008.
Artigo em Chinês | WPRIM | ID: wpr-270666

RESUMO

<p><b>OBJECTIVES</b>To investigate the effect of HOE642 on cardiac myocyte apoptosis of the heterotopic heart transplantation of rat non heart-beating donors.</p><p><b>METHODS</b>Totally 112 male Sprague-Dawley rats were randomly divided into 7 groups (n=16 in each group) C, the control group (normal hearts); S10, S30, and S45 (groups of transplanted hearts after 10, 30, and 45 minutes of asystole); and SH10, SH30, and SH45 (groups of transplanted hearts after 10, 30, and 45 minutes of asystole and infused with HOE642). After rata in the experimental groups were killed by warm ischemia the donators of the S10, S30 and S45 groups were infused with 5TH-1 for 30 minutes, and the dead rats in group SH10, SH30, and SH4 were infused with STH-1 and HOE642 (20 micromol/L) for 30 minutes. Heterotopic heart transplantation were processed by the method of neck Cuff. The heart specimens of S10, SH10, S30, and SH30 groups were taken after 48 hours of transplantation, and the heart specimens of S45 and SH45 groups were taken immediately after transplantation. Then apoptotic myocytes were detected with terminal deoxynucleotide transferase-mediated deoxyuridine-biotin nick end labeling method and the expressions of Bcl-2, Bax, and Caspase-3 proteins were detected by immunohistochemistry.</p><p><b>RESULTS</b>The rats were discerned death when cardiac electric wave vanished after 9-11 minutes of bloodletting by transsection of abdominal aorta. The number of positive cardiac muscle cells in S10 and S30 groups were significantly larger than those in group SH10 and SH30 (P < 0.05). The levels of Bcl-2 protein expression in S10 and S30 groups were significantly lower than those in SH10 and SH30 groups (P < 0.05). The levels of Bax and Caspase-3 protein expression were significantly higher than those in SH10 and SH30 groups (P < 0.05).</p><p><b>CONCLUSIONS</b>The rat model of a heterotopic heart transplantation on the cervical part is a convenient animal model for cardiac muscle protection. HOE642 can suppress rat cardiac muscle cells apoptosis (within 30 min) after death caused by warm ischemia.</p>


Assuntos
Animais , Masculino , Ratos , Antiarrítmicos , Farmacologia , Apoptose , Expressão Gênica , Guanidinas , Farmacologia , Coração , Transplante de Coração , Contração Miocárdica , Miócitos Cardíacos , Biologia Celular , Metabolismo , Proteínas Proto-Oncogênicas c-bcl-2 , Genética , Metabolismo , Distribuição Aleatória , Ratos Sprague-Dawley , Sulfonas , Farmacologia
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