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OBJECTIVE@#To study the trinucleotide repeats of GCN (GCA, GCT, GCC, GCG) encoding Alanine in exon 3 of the PHOX2B gene among healthy individuals from southwest China and two patients with Congenital central hypoventilation syndrome (CCHS).@*METHODS@#The number and sequence of the GCN repeats of the PHOX2B gene were analyzed by capillary electrophoresis, Sanger sequencing and cloning sequencing of 518 healthy individuals and two newborns with CCHS, respectively.@*RESULTS@#Among the 1036 alleles of the 518 healthy individuals, five alleles were identified, including (GCN)7, (GCN)13, (GCN)14, (GCN)15 and (GCN)20. The frequency of the (GCN)20 allele was the highest (94.79%). And five genotypes were identified, which included (GCN)7/(GCN)20, (GCN)13/(GCN)20, (GCN)14/(GCN)20, (GCN)15/(GCN)20, (GCN)20/(GCN)20. The homozygous genotypes were all (GCN)20/(GCN)20, and the carrier rate was 89.58%. Four GCN sequences of the (GCN)20 homozygous genotypes were identified among the 464 healthy individuals. The GCN repeat numbers in the exon 3 of the PHOX2B gene showed no significant difference between the expected and observed values, and had fulfilled the,Hardy-Weinberg equilibrium. The genotypes of the two CCHS patients were (GCN)20/(GCN)25 and (GCN)20/(GCN)30, respectively.@*CONCLUSION@#It is important to determine the GCN repeats and genotypic data of the exon 3 of the PHOX2B gene among the healthy individuals. The number of GCN repeats in 518 healthy individuals was all below 20. The selection of appropriate methods can accurately detect the polyalanine repeat mutations (PARMs) of the PHOX2B gene, which is conducive to the early diagnosis, intervention and treatment of CCHS.
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Humanos , Recién Nacido , Proteínas de Homeodominio/genética , Hipoventilación/congénito , Mutación , Apnea Central del Sueño/genética , Factores de Transcripción/genéticaRESUMEN
Parkinson's disease (PD) is a neurodegenerative disease characterized by selective loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc). We recently reported that Six2 could reverse the degeneration of DA neurons in a dephosphorylation state. Here we further identified that Eya1 was the phosphatase of Six2 that could dephosphorylate the tyrosine 129 (Y129) site by forming a complex with Six2 in damaged DA cells. Dephosphorylated Six2 then translocates from the cytoplasm to the nucleus. Using ChIP-qPCR and dual luciferase assay, we found that dephosphorylated Six2 down-regulates TEA domain1 (Tead1) expression, thus inhibiting 6-hydroxydopamine (6-OHDA)-induced apoptosis in DA cells. Furthermore, we showed Six2Y129F/Tead1 signaling could protect against the loss of SNpc tyrosine hydroxylase-positive (TH+) cells and improve motor function in PD model rats. Our results demonstrate a dephosphorylation-dependent mechanism of Six2 that restores the degeneration of DA neurons, which could represent a potential therapeutic target for PD.
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In the design of the outer windshield of high-speed trains, the non-fully enclosed windshield structure is widely used. Based on the cavity flow-induced vibration theory and FW-H acoustic equation, the flow field and acoustic characteristics of three high-speed train models with different windshield configurations are compared and analyzed. Numerical simulations were validated by full-scale experiments. The results show that the noise generated from the windshield area is mainly composed of low-frequency peak resonance noise and broadband turbulent noise. A typical Helmholtz resonator cavity is composed of inner and outer windshields. The position and number of windshield openings affect the airflow structure inside the windshield and the train wake. In addition, the hole as a potential aerodynamic noise source determines the vortex shedding and cavity resonance frequency, resulting in different acoustic feedback. The energy of windshield noise is mainly concentrated in the low frequency range below 400Hz. The windshield configuration of M1 and M3 shows tonal noise characteristics, while the M2 model eliminates the generation of low-frequency resonance noise. Compared with the M1 and M3 models, the noise generated by the windshield and the whole train of the M2 model is reduced by 3.83 dB and 1.77 dB, respectively.
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Acústica , Vibración , RuidoRESUMEN
Objective:To investigate the inhibitory effect of curcumin on oxidative stress in BCG-infected macrophages based on the Nrf2 pathway.Methods:THP-1-derived macrophages were infected.The experiment was divided into control group,BCG group,BCG+curcumin group and BCG+curcumin+ML385 group.Cellular ROS fluorescence intensity were observed under a fluores-cence microscope;Glutathione(GSH)levels were measured by Colorimetry;Western blot was used to detect the protein expressions of Nrf2,HO-1 and NQO1;MTT was used to detect the proliferation rate of macrophages.Results:BCG infection significantly enhanced ROS fluorescence intensity,reduced cell GSH content(P<0.01),inhibited protein expressions of Nrf2,HO-1 and NQO1,at the same time inhibited cell proliferation(P<0.01);curcumin significantly weakened ROS fluorescence intensity,increased GSH level(P<0.05),promoted Nrf2,HO-1 and NQO1 protein expressions and cell proliferation(P<0.01);Nrf2 inhibitor ML385 reversed the effect of curcumin.Conclusion:Curcumin can alleviate BCG-induced oxidative stress in macrophages by increasing the expression of Nrf2 and inducing the transcription of downstream antioxidant molecules.
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This study evaluated the scientific nature and effectiveness of iterative optimization of prevention and control measures for local outbreaks caused by the BA.2 and BA.5.2 COVID-19 strains in Fujian Province in 2022,to provide a scientif-ic basis for responding to future new or recurrent respiratory infectious diseases.According to the theory of infectious disease dynamics,relevant information regarding the local epidemic situation caused by the BA.2 sub-type Omicron virus strain in March 2022 and BA.5.2 sub-type Omicron virus strain in October 2022 in Fujian Province was collected.The susceptible exposed infectious removed(SEIAR)model of COVID-19 infection with a latent period and asymptomatic infected persons was used to analyze the transmission dynam-ics of two local epidemic situations,and evaluate the preven-tion and control effects.The incubation period of the BA.2 epidemic was 3 days(1~9 days),the intergenerational inter-val was 3 days(1~5 days),and the initial Rt was 3.0(95%CI:2.7~3.3).The incubation period of the BA.5.2 epidemic was 2 days(1~6 days),the intergenerational interval was 1 day(0~2 days),and the initial R,was 1.9(95%CI:1.7~2.1).The fittingresults for the BA.2 and BA.5.2 epidemics were good,and no statistical difference was observed between the predic-ted and actual numbers of cases(x2BA.2=31.53,x2BA.5.2=27.88,P>0.05).If an emergency response had not been initiated,the BA.2 epidemic would have continued to spread andpeak on April 7th,with an estimated 638 035 cases.The BA.5.2 epidemic would have rapidly spread,reaching a peak on November 14th,with an estimated 685 940 cases.If one incubation period were detected early,the scale of the BA.2 epidemic would have decreased by 25.73%;if two incubation periods were detected early,the scale would have decreased by 79.56%,and if one incubation period had been delayed,the scale would have expanded by 13.72%.If one incubation period had been detected early in the BA.5.2 epidemic,the scale would have decreased by 35.04%;if two incubation periods had been detected early,the scale would have decreased by 92.47%;and if one incubation period had been delayed,the scale would have increased by 19.75%.The guiding ideology,and the prevention and control measures for handling two local epidemics were optimized and iterated.Our study indicated that implementing the"four early"measures ef-fectively decreased the scale of the epidemic,and earlier detection was associated with more significant control effects.This study provides valuable information for the prevention and control of new or recurrent respiratory infectious diseases.
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Dangefentong Capsules is a new traditional Chinese medicine preparation for the treatment of diabetic peripheral neuropathy. It is based on the Salviae Miltiorrhizae Radix et Rhizoma-Puerariae Lobatae Radix herb pair with salvianolic acids, tanshinones and pueraria flavonoids as main components. Studying the chemical composition in vivo of Dangefentong Capsules and its metabolites is of great significance for making clear its pharmacodynamic material basis and the action mechanism. The UHPLC-Q/Orbitrap-MS/MS was applied to rapidly analyze the metabolites and metabolic pathways of Dangefentong Capsules in Beagle dogs after gavage. Eclipse plus C_(18) column(2.1 mm×50 mm, 1.8 µm) was used, and gradient elution was performed with 0.1% formic acid aqueous solution(A)-formic acid acetonitrile solution(B). A heated electrospray ion source(HESI) was employed. The scanning mode was set as the positive and negative ion mode, and the mass scanning range was m/z 100-1 000. The plasma, urine and feces samples were collected after male Beagle dogs were administered with Dangefentong Capsules. The prototype components and metabolites were identified by UHPLC-Q/Orbitrap-MS/MS analysis combined with reference substances and references. The results showed that 107 chemical components were identified, including 58 prototype components and 49 metabolites. The identified prototype components included 42 components from Salviae Miltiorrhizae Radix et Rhizoma and 16 components from Puerariae Lobatae Radix. The metabolites consist of 21 and 28 metabolites of Salviae Miltiorrhizae Radix et Rhizoma and Puerariae Lobatae Radix, respectively. They are mainly derived from the methylation, hydroxylation, sulfation and glucuronidation of salvianolic acids, tanshinones and pueraria flavonoids. This research rapi-dly analyzes the chemical components in vivo of Beagle dogs administered with Dangefentong Capsules, laying a basis for illustrating the pharmacodynamic material basis and mechanism of Dangefentong Capsules.
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Medicamentos Herbarios Chinos , Pueraria , Abietanos , Acetonitrilos , Alquenos , Animales , Cápsulas , Cromatografía Líquida de Alta Presión/métodos , Perros , Medicamentos Herbarios Chinos/química , Flavonoides , Formiatos , Masculino , Polifenoles , Espectrometría de Masas en TándemRESUMEN
It is still difficult to conduct numerical calculation of the aerodynamic noise of full-scale, long-marshalling, high-speed trains. Based on the Lighthill acoustic analogy theory, the aerodynamic sound source of the high-speed train is equivalent to countless micro-vibrating sound sources. An acoustic radiation model of the dipole sound source of high-speed trains is established, and a method to predict the aerodynamic noise in the far field of long-marshalling high-speed trains is proposed. By this method, combined with numerical simulation technology, the flow field, noise source, and far-field noise characteristics of high-speed trains with different marshalling numbers are studied. The improved delayed detached eddy simulation method is used for flow field calculation, to obtain aerodynamic noise source information regarding the surface of high-speed trains. The numerical calculation method is verified by wind tunnel testing. The results show that the flow field and noise source characteristics of high-speed trains with different marshalling numbers are similar. The greater the length of the train body, the longer the trailing distance of the train wake, and the stronger of a surface noise source the tail car becomes. The spatial distribution characteristics of aerodynamic noise in the far field of high-speed trains do not change significantly with the length of the train body, but the magnitude of the sound pressure level will increase with the increase in length of the train body. The middle car body parts of high-speed trains with different marshalling numbers have similar noise distributions and sound pressure levels. Based on the noise calculation results of the 3-marshalling high-speed train, the far-field noise of the 5-marshalling and 8-marshalling train models is predicted and found to be in good agreement with the far-field noise of the actual train model. The differences in average sound pressure level are 1.01 dBA and 1.74 dBA, respectively.
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Glial cell line-derived neurotrophic factor (GDNF) played critical roles in the survival and repair of dopaminergic (DA) neurons. Transcription factor Six2 could repair injured DA cells by promoting the expression of GDNF, however, the underlying molecular mechanisms remain largely unknown. In this study, we screened forty-three proteins that interacted with Six2 in MES23.5 DA cells treated with 6-OHDA by liquid chromatography - electrospray - ionization tandem mass spectrometry (LC-ESI-ITMS/MS). Among these proteins, Smarcd1 is a member of SWI/SNF chromatin-remodeling complex family. Our results confirmed that Smarcd1 formed a transcription complex with Six2, and Smarcd1 mainly binded to the 2840 bp-2933 bp region of the GDNF promoter. Furthermore, knockdown of Smarcd1 inhibited the effect of Six2 on GDNF expression, and resulted in decreased cell viability and increased the apoptosis of injured DA neurons, and the result of overexpression of Smarcd1 is opposite to knockdown. Taken together, our results indicate that smarcd1 can be recruited to the promoter region of GDNF by transcription factor Six2 to promote the effect of Six2 on GDNF expression and protect injured MES23.5 DA cells, which could be useful in identifying potential drug targets for promoting endogenous GDNF expression.
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Neuronas Dopaminérgicas/metabolismo , Factor Neurotrófico Derivado de la Línea Celular Glial/genética , Proteínas de Homeodominio/metabolismo , Factores de Transcripción/metabolismo , Animales , Apoptosis , Línea Celular , Neuronas Dopaminérgicas/patología , Regulación de la Expresión Génica , Células Híbridas , Ratones , RatasRESUMEN
Objective:To investigate the role of peroxisome proliferator-activated receptor γ (PPARγ)/CD36 signaling pathway in macrophage lipid metabolism after Mycobacterium tuberculosis ( Mtb) infection. Methods:THP-1-derived macrophages were infected with Mtb. Four groups were included in this study, which were control group, Mtb infection group, Mtb+ rosiglitazone (ROZ, PPARγ agonist) group and Mtb+ GW9662 (PPARγ antagonist) group. Western blot and RT-PCR were used to detect the expression of PPARγ in macrophages at protein and mRNA levels, respectively. The lipids in cells were detected by oil red O staining. The concentrations of total cholesterol (TC), triglycerides (TG), low density lipoprotein (LDL-C) and high density lipoprotein (HDL-C) in the supernatant of cell culture were detected by automatic biochemical analyzer. The expression of CD36 was detected by immunohistochemistry. CCK-8 was used to detect the proliferation rate of macrophages. Results:Mtb infection significantly increased the expression of PPARγ in macrophages ( P<0.001), promoted intracellular lipid aggregation and CD36 expression and decreased the levels of TC, TG, LDL-C and HDL-C in the supernatant of cell culture ( P<0.001) and cell proliferation rate ( P<0.001). PPARγ agonist significantly enhanced the intracellular lipid accumulation and CD36 expression that were induced by Mtb infection and down-regulated the lipid level in the supernatant of cell culture and cell proliferation rate, while PPARγ antagonist reversed the above effects. Conclusions:PPARγ played a role in lipid metabolism in Mtb-infected macrophages through affecting CD36 expression.
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Objective To evaluate the effects of antiretroviral therapy(ART)for the prevention of mother-to-child transmission(PMTCT)of acquired immune deficiency syndrome(AIDS)on the growth and development of 18-month-old children born by human immunodeficiency virus(HIV)-positive pregnant women in Lingshan County,Guangxi Zhuang Autonomous Region,and provide scientific evidence for improving the ART medication plan for PMTCT.Methods Lingshan County,ranking the first in the HIV-epidemic counties of Guangxi,was selected as the research site.According to the design of retrospective case-control study,we assigned all the subjects into the case group and the control group:(1)The case group included the HIV-positive pregnant women who had received ART for PMTCT and their HIV-negative infants in Lingshan County from 2010 to 2017.The historical cards and PMTCT data of them were collected from the national PMTCT database.(2)The control group included the healthy pregnant women and their healthy babies born in the Lingshan Maternity and Infant Hospital in 2017,and the children's growth and development data were collected.The stunted growth in children was defined as at least one of the three main indicators of body height,body weight,and head circumference below the normal range.Results The number of HIV-positive mothers and their infants in the case group was 391 and 368,respectively,and 87.21%(341/391)and 95.38%(351/368)of mothers and infants respectively received ART medication.The HIV positive rate,mortality rate,and mother-to-child transmission rate of 18-month-old children were 1.36%(5/368),4.35%(16/368),and 2.01%(5/249),respectively.The incidence of stunted growth of 18-month-old children in the case group and the control group was 42.12%(155/368)and 23.06%(101/438),respectively,with significant difference(
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Femenino , Humanos , Lactante , Embarazo , Estudios de Casos y Controles , China/epidemiología , Crecimiento y Desarrollo , VIH , Infecciones por VIH/prevención & control , Transmisión Vertical de Enfermedad Infecciosa/prevención & control , Madres , Complicaciones Infecciosas del Embarazo/tratamiento farmacológico , Estudios RetrospectivosRESUMEN
Objective: To investigate the effect of Bifidobacterium on endothelial nitric oxide synthase and its potential protective mechanism in neonatal necrotizing enterocolitis using an induced mouse model. Methods: Sixty 8-day newborn C57BL/6 mice were randomized into three groups (n=20): control group, necrotizing enterocolitis group (NEC) and Bifidobacterium intervention group (NEC+BIF). Samples were collected 3 days after NEC modeling. HE staining pathological score was used to evaluate the modeling effect; the level of endothelial nitric oxide synthase (eNOS) in the intestine was detected with Western blotting; the eNOS activity, nitric oxide (NO) and superoxide (O2•) content were measured by Colorimetry. Real-time quantitative PCR was used to detect the mRNA level of interleukin-6 (IL-6) and interleukin-1β (IL-1β); the protein expression of IL-6 and IL-1β was assessed by ELISA. Results: No significant difference was observed in the protein levels of eNOS between the three groups. Compared with control group, in NEC group, the eNOS activity and NO content were significantly decreased; the O2 content was substantially increased; both IL-6 and IL-1β were dramatically increased (P0.05). Conclusion: Bifidobacterium can alleviate the severity of intestinal damage in NEC mouse model, the mechanism may be related to the enhancement of eNOS activity.
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A novel, rapid and sensitive chemiluminescence (CL) method for the determination of oxytetracycline hydrochloride (OTCH) is described in this paper. The presented method was based on the fact that OTCH could immensely enhance the CL of the reaction of cerium sulfate and tris(2,2-bipyridyl) ruthenium (II) in acidic medium. Under optimal experimental conditions, CL intensity was favorably linear for OTCH in the range 5.0 × 10-7 to 5.0 × 10-5 g/ml, with a detection limit of 1.5 × 10-7 g/ml (S/N = 3). The relative standard detection was 4.76% for 5.0 × 10-6 g/ml (n = 11). This method was successfully applied to the analysis of OTCH in milk and egg white samples. According to the results of the kinetic curves for OTCH in the Ru(bipy)3 2+ -Ce(SO4 )2 CL system, together with CL and ultraviolet (UV)-visible spectra, the possible mechanism of the CL reaction is discussed briefly.
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Antibacterianos/análisis , Clara de Huevo/análisis , Mediciones Luminiscentes/métodos , Leche/química , Oxitetraciclina/análisis , Animales , Bovinos , Cerio/química , Pollos , Contaminación de Alimentos/análisis , Mediciones Luminiscentes/instrumentación , Rutenio/química , Sulfatos/químicaRESUMEN
Objective:To observe the clinical efficacy of balance-impact tuina therapy for lumbar intervertebral disc herniation (LIDH).Methods:A total of 118 eligible LIDH patients were randomized into an observation group and a control group by the random number table method,with 59 cases in each group.The observation group was intervened by balance-impact tuina therapy,while the control group was intervened by conventional tuina therapy,both for consecutive two weeks.The scores of visual analog scale (VAS),Oswestry disability index (ODI),quality of life questionnaire-core 30 (QOL-C30) were observed before and after treatment;the relapse rate was estimated at the sixth month and twelfth month following the treatment.The data were statistically analyzed.Results:After intervention,the total effective rate was 96.6% in the observation group versus 91.5% in the control group,and the between-group difference was statistically significant (P<0.05).The VAS and ODI scores declined significantly after treatment in both groups (all P<0.05),and the observation group was markedly lower than the control group (P<0.05,P<0.01).The QOL-C30 score increased significantly after treatment in both groups (both P<0.05),and the observation group was markedly higher than the control group (P<0.05).The relapse rates at the post-treatment sixth month and twelfth month in the observation group were lower than those in the control group (P<0.05,P<0.01).Conclusion:Compared with the conventional tuina therapy,the balance-impact tuina therapy shows advantage in lessening pain,improving the function and enhancing the quality of life in the treatment of LIDH,and it has a lower relapse rate.Thus,this therapy is worth promoting in clinic.
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Objective:To investigate evaluation methods to predict the outcome of nervous system development in high-risk infants. Methods:From March, 2015 to March, 2016, 336 high-risk infants were enrolled. They were assessed by General Movements (GMs) Quality Assessment, 0~1 Years Old 20 Items Neuromotor Assessment and Gesell Developmental Schedules. Results:A total of 236 infants finishied the study. GMs Quality Assessment showed that 203 cases were normal and 33 cases were abnormal in the writhing movements stage; 218 cases were normal and 18 cases were abnormal in the fidgety movemonts stage. 0~1 Years Old 20 Items Neuromotor Assessment showed that 202 cases were normal and 34 cases were abnormal. Gesell Developmental Schedules showed that 12 cases were abnormal. Conclusion:The combination of GMs Quality Assessment, 0~1 Years Old 20 Items Neuromotor Assessment and Gesell Developmental Schedules could better predict the nervous system development of high-risk infants.
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To follow up and analyze patients with ventricular septal rupture( VSR) after acute myocardial infarction ( AM I) w ho underwent VSR occlusion ,screen the main risk factors of survival in perioperative patients with VSR after AM I ,and observe the postoperative cardiac function and hemodynamic changes by echocardiography . Methods Seventeen VSR patients were divided into the survival group ( 11 cases) and the death group ( 6 cases) within 30 days according to the survival time . T he changes of cardiac ultrasound parameters before and after the operation of VSR survivors were compared , and the hemodynamic recovery characteristics of VSR survivors were analyzed . Results Age ,the time to VSR occlusion ,preoperative left ventricular ejection fraction ,cardiogenic shock ,and the size of VSR were all the factors that were closely related to the 30‐day mortality of VSR with odds ratio of 0 .90 ( 95% CI 0 .73 to 1 .14 , P =0 .045) ,1 .89( 95% CI 1 .35 to 2 .23 , P =0 .003) ,0 .89( 95% CI 0 .57 to 1 .24 , P =0 .039) ,1 .45 ( 95% CI 1 .12 to 1 .78 , P =0 .027) and 11 .45( 95% CI 7 .89 to 15 .56 , P =0 .012) ,respectively ( all P<0 .05 ) . Compared with the preoperative measurements , the left ventricular end‐diastolic volume and pulmonary artery systolic pressure were significantly reduced in the VSR survival group ( P < 0 .05 ) . Conclusions The decrease of left ventricular end diastolic volume and pulmonary artery systolic pressure after operation indicates a better prognosis in the short period . Echocardiography is a vital tool in preoperative screening ,intraoperative monitoring and postoperative follow‐up in VSR occlusion .
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miR-214 plays a major role in the self-renewal of skin tissue. However, whether miR-214 regulates the proliferation and differentiation of human hair follicle stem cells (HFSCs) is unknown. Primary HFSCs were isolated from human scalp skin tissue, cultured, and identified using flow cytometry. An miR-214 mimic and inhibitor were constructed for transfection into HFSCs. The MTS and colony formation assays examined cell proliferation. Immunofluorescence detected the localization and expression levels of TCF4, β-catenin, and differentiation markers. Luciferase reporter and TOP/FOP Flash assays investigated whether miR-214 targeted EZH2 and regulated the Wnt/β-catenin signaling pathway. Western blot determined the expression levels of enhancer of zeste homolog 2 (EZH2), Wnt/β-catenin signaling-related proteins, and HFSC differentiation markers in cells subjected to miR-214 transfection. miR-214 expression was remarkably decreased during the proliferation and differentiation of HFSCs into transit-amplifying (TA) cells. Downregulation of miR-214 promotes the proliferation and differentiation of HFSCs. Overexpression of miR-214 led to decreased expression of EZH2, β-catenin, and TCF-4, whereas downregulation of miR-214 resulted in increased expression of EZH2, β-catenin, and TCF-4 as well as TA differentiation markers. Immunofluorescence assay revealed that inhibiting miR-214 triggered the entry of β-catenin and TCF-4 into the nucleus. The luciferase reporter and TOP/FOP Flash assays demonstrated that miR-214 directly targets EZH2 and affects Wnt/β-catenin signaling. The miR-214/EZH2/β-catenin axis could be considered a candidate target in tissue engineering and regenerative medicine for HFSCs.
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Humanos , Antígenos de Diferenciación , Western Blotting , Proliferación Celular , Regulación hacia Abajo , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Folículo Piloso , Cabello , Técnicas In Vitro , Luciferasas , Medicina Regenerativa , Cuero Cabelludo , Piel , Células Madre , Ingeniería de Tejidos , TransfecciónRESUMEN
Objective To study the effect of an essential α,β-unsaturated aldehyde from cigarette smoke crotonal-dehyde on myocardial contractile function and intracellular Ca2+function in mice. Methods Hearts of from male C57BL/6 mice were digested by Langendorff to islate the cardiomyocytes. The cardiomyocytes of mice were then in-cubated with crotonaldehyde(1,10,25 and 50 μmol/L) for 6 h,and the control group was treated without croton-aldehyde,then they were evaluated including peak shortening(PS),maximal velocity of shortening/relengthening (±dL/dt),time-to-PS(TPS),time-to-90% relengthening(TR90),fura-2 fluorescence intensity(FFI),intracel-lular Ca2+decay and SERCA 45Ca2+uptake and the expression of Na+-Ca2+exchange were evaluated by Western blot analysis. Results Compared with the control group, the higher concentrations of the crotonaldehyde(25 and 50 μmol/L) groups significantly diminished the PS, ±dL/dt,ΔFFI,SERCA activity and Ca2+decay (P<0.05), as well as prolonged the TR90(P<0.05);however the crotonaldehyde with different concentrations had no effect on the expression of Na+-Ca2+exchange in cardiomyocytes. Conclusions Crotonaldehyde may inhibit cardiomyocyte contraction by suppressing SERCA activity and compromising intracellular Ca2+handling.
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· AIM:To explore the modulation effects of 17-β estradiol (E2) and tamoxifen (TAM) in chronic intraocular hypertension mouse model.· METHODS:We performed anterior chamber injection of magnetic beads to induced chronic ocular hypertension models.Adult C57BL/6 male mice were used in the experiments and randomly divided into four groups:control,Beads group,E2 group and E2+TAM group.The intraocular pressure (lOP) were measured by Tonolab tonometer.Central retinal thickness was evaluated by HE staining.Brn3a as a specific marker of retinal ganglion cells (RGCs),were stained and counted by immunohistochemistry (IHC) staining.Glial fibrillary acidic protein (GFAP) as a marker of proliferation of astrocytes,was quantified using western blotting.· RESULTS:The IOP level was significantly elevated after anterior chamber injection of magnetic beads compared to control group (P<0.05),while in E2 and E2+TAM group,the IOP levels were reduced (P< 0.05 vs Beads group),especially in E2+TAM group in 2wk.The RGCs happened to degenerated in Beads group after 2wk,while the effects were reversed by E2+TAM (P<0.05).The central retinal thickness showed no significant statistical difference among the four groups after 2wk (P > 0.05).The expression level of GFAP increased caused via beads injection,however,decreased in E2 and E2+TAM group (P<0.05).· CONCLUSION:E2 and E2+TAM could both effectively decrease the IOP level in chronic intraocular hypertension mouse model,increase the survival of RGCs from high intraocular pressure,suppress expression of GFAP,which indicated neuroprotective effects of E2 and TAM in glaucoma through an anti-inflammatory effects.
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Objective To investigate the effects of dedicator of cytokinesis 1 (Dock180) knockout on proliferation and apoptosis in rat derived H9C2 cardiomyocytes and their mechanisms.Methods A single guide RNA (sgRNA) targeting rat Dock180 gene was designed and constructed using CRISPR/Cas9 system.A plasmid contained above sgRNA was packaged into lentivirus and selected to knockout Dock180 in the cardiomyocytes.A single clone of cardiomyocyte with Dock180 knockout was established.Cardiomyocytes were divided into negative lentivirus group (Cas9, A group), Dock180 knockout group (B group), Cas9 lentivirus hypoxia group (C group), Dock180 knockout hypoxia group (D group).The expression of Dock180 mRNA was examined by RT-PCR, and relevant proteins were detected by Western blot.The cell proliferation rate of the cardiomyocytes was determined by MTT, and the apoptotic rate was measured by flow cytometry.Results Dock180 mRNA and protein were absent in B andD groups.Compared with A and C groups, p-ERK1/2 and Bcl-2 protein expression and cell proliferation rate were lower in B and D groups respectively (P<0.01), while Bax protein expression and cell apoptosis rate were higher in B and D groups respectively (P<0.05, P<0.01);Compared with A group, Dock180 mRNA and protein, p-ERK1/2 and Bcl-2 proteins and cell proliferation rate were reduced, while Bax protein and cell apoptosis rate were increased in C group(P<0.05,P<0.01).Compared with B group, p-ERK1/2 and Bcl-2 proteins and cell proliferation rate were decreased, while Bax protein and cell apoptosis rate were increased in D group(P<0.05,P<0.01).Conclusions Dock180 knockout with CRISPR/Cas9 can inhibit proliferation and promote apoptosis via p-ERK1/2, Bcl-2 and Bax in H9C2 cardiomyocytes.
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The aim of this study was to investigate the relationship between the acetylcholine concentration in the blood and gelsenicine-induced death in mice. Kunming mice were given intraperitoneal injections of normal saline, gelsenicine or different doses of acetylcholine chloride. Atropine was given to the mice which received gelsenicine or medium dose acetylcholine chloride injection. The blood was sampled immediately when the mice died or survived for 20 min after injection. The acetylcholine concentration and acetylcholinesterase activity in the blood were measured by the testing kits, and the mortality was calculated and analyzed. The results showed that half lethal dose of gelsenicine (0.15 mg/kg) reduced the acetylcholinesterase activity and increased the blood acetylcholine concentration. The blood acetylcholine concentration of the dead mice in the gelsenicine group was increased to 43.0 μg/mL (from 31.1 μg/mL in the control), which was lower than that (53.9 μg/mL) of the dead mice in the medium dose acetylcholine chloride group, but almost equal to that (42.7 μg/mL) of the survival mice in the medium dose acetylcholine chloride group. Atropine could successfully rescue the mice from acetylcholine poisoning, but its efficiency of rescuing the mice from gelsenicine intoxication was weak. These results suggest that gelsenicine can inhibit acetylcholinesterase activity and increase blood acetylcholine concentration, but the accumulation of acetylcholine may not be the only or main cause of the death induced by gelsenicine in mice.