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1.
Zhongguo Zhong Yao Za Zhi ; 48(20): 5548-5557, 2023 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-38114147

RESUMO

To explore the quality consistency evaluation method for multi-component traditional Chinese medicine and establish a dissolution evaluation method suitable for the characteristics of multi-component Chinese patent medicine, this study discussed the characteristics and advantages of the flow-through cell method in the dissolution evaluation of Chinese patent medicine by comparing the impact of the small cup method and the flow-through cell method on the dissolution behavior of water-soluble and lipid-soluble major active components of Danshen Tablets. Dissolution tests were performed using the small cup method as described in the 2020 edition of the Chinese Pharmacopoeia and the newly introduced flow-through cell method(closed-loop method) with water solution containing 0.5% SDS as dissolution medium. Cumulative dissolution curves of the water-soluble component salvianolic acid B and the lipid-soluble component tanshinone Ⅱ_A in Danshen Tablets were plotted, and fitting and similarity analysis of the dissolution models was conducted to identify the characteristics and advantages of the flow-through cell method. For the small cup method, 150 mL of water containing 0.5% SDS was used as the dissolution medium, with a rotation speed of 75 r·min~(-1) and a temperature of(37±0.5) ℃, and 3 mL of samples were taken at 15, 30 min, 1, 2, and 4 h, with fresh dissolution medium added at the same temperature and volume. For the flow-through cell method, a closed-loop system was used. Danshen Tablets were placed in the flow-through cell with approximately 6.7 g of glass beads, and 150 mL of water containing 0.5% SDS was used as the dissolution medium. The flow rate was set at 20 mL·min~(-1), and the temperature and sampling were the same as the small cup method. The results showed that compared with the small cup method, the flow-through cell method had stronger discriminative power and higher sensitivity in distinguishing the dissolution behavior of the two components, and could better reflect the differences in formulation quality, especially for water-insoluble lipid-soluble components. Given that there were no essential differences in the in vitro release kinetics between the two methods, the flow-through cell method could not only replace the traditional small cup method but also better guide the formulation development and identify quality issues of formulations.


Assuntos
Salvia miltiorrhiza , Medicina Tradicional Chinesa , Comprimidos , Água , Lipídeos , Solubilidade
2.
Artigo em Chinês | WPRIM | ID: wpr-981355

RESUMO

This study aimed to explore the correlation between traditional Chinese medicine(TCM) and reduced risk of readmission in patients having rheumatoid arthritis with hypoproteinemia(RA-H). A retrospective cohort study was conducted on 2 437 rheumatoid arthritis patients in the information system database of the First Affiliated Hospital of Anhui University of Chinese Medicine from 2014 to 2021, and 476 of them were found to have hypoproteinemia. The patients were divided into TCM users and non-TCM users by propensity score matching. Exposure was defined as the use of oral Chinese patent medicine or herbal decoction for ≥1 month. Cox regression analysis was performed to explore the risk factors of clinical indicators of rheumatoid arthritis. Additionally, the use of TCM during hospitalization was analyzed, and analysis of association rules was conducted to investigate the correlation between TCM, improvement of indicators and readmission of patients. Kaplan-Meier survival curve was plotted to compare the readmission rate of TCM users and non-TCM users. It was found the readmission rate of RA-H patients was significantly higher than that of RA patients. By propensity score matching, 232 RA-H patients were divided into TCM group(116 cases) and non-TCM group(116 cases). Compared with the conditions in the non-TCM group, the readmission rate of the TCM group was lowered(P<0.01), and the readmission rate of middle-aged and elderly patients was higher than that of young patients(P<0.01). Old age was a risk factor for readmission of RA-H patients, while TCM, albumin(ALB) and total protein(TP) were the protective factors. During hospitalization, the TCMs used for RA-H patients were mainly divided into types of activating blood and resolving stasis, relaxing sinew and dredging collaterals, clearing heat and detoxifying, and invigorating spleen and resolving dampness. The improvement of rheumatoid factor(RF), immunoglobulin G(IgG), erythrocyte sedimentation rate(ESR), C-reactive protein(CRP) and ALB was closely related to TCM. On the basis of western medicine treatment, the application of TCM could reduce the readmission rate of RA-H patients, and longer use of TCM indicated lower readmission rate.


Assuntos
Pessoa de Meia-Idade , Idoso , Humanos , Medicina Tradicional Chinesa , Medicamentos de Ervas Chinesas/uso terapêutico , Estudos Retrospectivos , Readmissão do Paciente , Artrite Reumatoide/tratamento farmacológico , Hipoproteinemia/tratamento farmacológico
3.
Journal of Modern Urology ; (12): 770-774, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1005991

RESUMO

【Objective】 To analyze the correlation between CT attenuation value of renal papilla and occurrence and development of renal calculi. 【Methods】 The clinical data of 100 patients with calcium oxalate stones treated during Aug.2020 and Jul.2022 were retrospectively analyzed, including 60 with primary stones, and 40 with recurrent stones. The 30 healthy volunteers were enrolled in the control group. CT attenuation value of renal papilla was measured with plain scan CT, and risk factors were identified with the receiver operating characteristic (ROC) curve. The correlation of CT attenuation value and 24 h urine metabolism was analyzed. 【Results】 The CT attenuation value of renal papilla was significantly higher in the primary group [34.92 (IQR: 3.84)] and recurrent group [43.00 (IQR: 8.74)] than in the control group [32.58 (IQR: 5.21)] (P<0.05). Compared with the primary group, the recurrent group had decreased citric acid level but increased calcium level in 24 h urine (P<0.05). The citrate ion and calcium ion were correlated with the CT attenuation value (P<0.05). 【Conclusion】 Patients with high renal papilla density have a high risk of stone formation and recurrence. Increased renal papilla density is a warning signal for the development of stones. The high calcium and low citric acid in 24 h urine have certain effects on the occurrence and development of urolithiasis. Intake of calcium should be limited and citric acid should be supplemented in patients with calcium oxalate stones.

4.
Zhongguo Zhong Yao Za Zhi ; 48(24): 6635-6644, 2023 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-38212023

RESUMO

To explore the stability characteristics of ß-nicotinamide mononucleotide(NMN) and provide data support for NMN production, preparation, and related product development, this study established a simple HPLC content determination method for NMN in simple substrate and investigated the degradation behavior, degradation products, and degradation kinetics of NMN under various chemical, physical, and biological conditions. The HPLC method employed a Welch Xtimate AQ-C_(18) column(4.6 mm×250 mm, 5 µm), a detection wavelength of 266 nm, a column temperature of 30 ℃, a flow rate of 1.0 mL·min~(-1), an injection volume of 5 µL, and a mobile phase consisting of methanol(A) and a 10 mmol·L~(-1) ammonium formate aqueous solution(B) with a gradient elution(0-6.7 min, 0-4% A; 6.7-13 min, 4%-18% A; 13-14.2 min, 18% A; 14.2-15 min, 18%-0 A; 15-22 min, 0 A). This method provided good separation between NMN and potential impurities and degradation products, and had a wide linear range, short analysis time, good durability, high accuracy, an average sample recovery rate of 98.71%, and an RSD of 1.2%. The instrument precision had an RSD of 0.26%, and the linearity within the examined range was excellent(R~2≥0.999 9). This method can be applied for NMN content determination in simple substrate. The degradation process of NMN in aqueous solution followed apparent first-order kinetics, with the degradation rate primarily influenced by high temperature and pH. NMN was more stable in low-temperature, neutral, or weakly acidic/alkaline environments. Strong acids or strong alkalis could accelerate its degradation, and its degradation rate was less affected by pepsin and trypsin. In an aqueous solution at room temperature, it followed the kinetic equation lg C_t=0.005 7t + 4.817 2, with t_(0.9) and t_(1/2) values of 95.58, 860.26 h, respectively. The results suggest that pH and temperature are the main factors affecting the stability of NMN in aqueous solution, and low temperature, moisture protection, and a weakly acidic environment are more conducive to the storage and application of NMN and its products.


Assuntos
Ácidos , Mononucleotídeo de Nicotinamida , Mononucleotídeo de Nicotinamida/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Cinética
5.
Journal of Biomedical Engineering ; (6): 1158-1164, 2022.
Artigo em Chinês | WPRIM | ID: wpr-970654

RESUMO

This study aimed to investigate the effect of curcumin (Cur) against human cytomegalovirus (HCMV) in vitro. Human embryonic lung fibroblasts were cultured in vitro. The tetrazolium salt (MTS) method was used to detect the effects of Cur on cell viability. The cells were divided into control group, HCMV group, HCMV + (PFA) group and HCMV + Cur group in this study. The cytopathic effect (CPE) of each group was observed by plaque test, then the copy number of HCMV DNA in each group was detected by quantitative polymerase chain reaction (qPCR), and the expression of HCMV proteins in different sequence was detected by Western blot. The results showed that when the concentration of Cur was not higher than 15 μmol/L, there was no significant change in cell growth and viability in the Cur group compared with the control group (P>0.05). After the cells were infected by HCMV for 5 d, the cells began to show CPE, and the number of plaques increased with time. Pretreatment with Cur significantly reduced CPE in a dose-dependent manner. After the cells were infected by HCMV, the DNA copy number and protein expression gradually increased in a time-dependent manner. Pretreatment with Cur significantly inhibited HCMV DNA copies and downregulate HCMV protein expression levels in a concentration-dependent manner, and the difference was statistically significant (P<0.05). In conclusion, Cur may exert anti-HCMV activity by inhibiting the replication of HCMV DNA and down-regulating the expression levels of different sequence proteins of HCMV. This study provides a new experimental basis for the development of anti-HCMV infectious drugs.


Assuntos
Humanos , Curcumina/uso terapêutico , Citomegalovirus/genética , Infecções por Citomegalovirus/tratamento farmacológico , Placa Aterosclerótica
6.
Artigo em Chinês | WPRIM | ID: wpr-885523

RESUMO

Objective:To investigate the levels of periphreal blood free carnitine and amino acids in healthy pregnant women in the third trimester and their association with maternal, fetal, and neonatal cardiac function and structure.Methods:This prospective descriptive study included healthy singleton pregnancies who underwent routine obstetric examination and delivered in two district maternal and child health hospitals (one in the urban and one in the suburb an area) in Beijing from June 2017 to February 2018. All recruiters had serology Down's syndrome screening test at (18±1) gestational weeks. Besides measurement of amino acids and free carnitine levels in whole blood and urine samples by liquid chromatography-tandem mass spectrometry, all cases underwent maternal and fetal echocardiography at (35±1) weeks of gestation. And neonatal echocardiography was performed after delivery to assess the heart function and structure. Antenatal factors were also collected, including maternal education background, age at first marriage and conception, gravidity, and folic acid supplement in early pregnancy. Statistical analysis was performed using t-test, ANOVA, Chi-square test, Pearson correlation coefficient, and Kappa test. Results:A total of 493 mother-neonate dyads were enrolled in this study. Blood free carnitine levels in the healthy pregnant women in the third trimester ranged from 5.09 to 59.17 μmol/L (reference value: 10.00-50.00 μmol/L) with an average value of (13.03±3.87) μmol/L. None was found with structural abnormalities by cardiac ultrasound, showing an average left ventricular end diastolic diameter (LVEDD) and end systolic diameter (LVESD) of (45.70±3.08) mm and (29.17±3.12) mm, respectively, and left ventricular ejection fraction (LVEF) of all cases were over 55%. No cardiac malformation was detected by the third-trimester fetal echocardiography. The average birth weight of the 493 newborns was (3 340±313) g. Those whose birth weight <2 500 g and >4 000 g were accounted for 1.0% (5 cases) and 3.0% (15 cases) with the average maternal blood free carnitine level of (13.25±2.17) μmol/L (10.46-19.21 μmol/L) and (12.64±2.50) μmol/L (8.78-17.73 μmol/L) ( t=0.42, P>0.05). The average LVEDD and LVESD of the 493 newborns were (17.21±1.27) mm and (11.03±1.30) mm, respectively. For the 64 newborns (13.0%) whose LVEF<60%, the maternal blood free carnitine level was (12.93±2.78) μmol/L (7.34-22.13 μmol/L), showing no statistical difference ( t=-0.29, P>0.05) with those 59 neonates (12.0%) whose LVEF over 75% and maternal carnitine level of (13.09±3.24) μmol/L (8.66-27.49 μmol/L). All cases were divided into four groups based on the quartiles of maternal blood free carnitine level and no significant difference in maternal or neonatal LVEDD or LVEF was observed among these groups (all P>0.05). Conclusions:Blood free carnitine concentration in healthy pregnant women in the third trimester is at the lower limit of normal range, and no significant effect on maternal cardiac function and fetal cardiac structure is seen. However, the effect of low maternal carnitine level in the third trimester on children's myocardial function and whether carnitine should be supplemented in the third trimester are worthy of further investigation with larger sample size.

7.
Fitoterapia ; 121: 159-163, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28739177

RESUMO

A phytochemical investigation on the 70% EtOH extract of the bark of Dysoxylum hainanense resulted in the isolation of four new triterpenoids, dysoxyhaines A-D (1-4). Structural elucidation of all the compounds were performed by spectral methods such as 1D and 2D (1H1H COSY, HMQC, and HMBC) NMR spectroscopy, in addition to high resolution mass spectrometry. The isolated components were evaluated in vitro for anti-inflammatory activities for Cox-1 and Cox-2, and radical scavenging potential using ABTS·+ and DPPH test. As a result, nor seco-olean type triterpenoid 1 exhibited significant anti-inflammatory potential, while tirucallane triterpenoids 3 and 4 showed radical scavenging activities.


Assuntos
Anti-Inflamatórios/química , Sequestradores de Radicais Livres/química , Meliaceae/química , Triterpenos/química , Anti-Inflamatórios/isolamento & purificação , Sequestradores de Radicais Livres/isolamento & purificação , Estrutura Molecular , Casca de Planta/química , Triterpenos/isolamento & purificação
8.
Artigo em Chinês | WPRIM | ID: wpr-230052

RESUMO

Vertebral artery type cervical spondylosis is a common type of cervical spondylosis, also known as cervical vertigo. In recent years, with the increase of people's life pace and the number of office staff, the incidence rate has been rising year by year, with a trend of younger age. Because traditional Chinese medicine has unique advantages in treating cervical vertigo, it is necessary to study the regularity of the Chinese medicine for cervical vertigo. But at present, the research is mainly based on the statistics for frequency and proportion of herbs. In addition, in the process of diagnosis and treatment of Chinese medicine, personal experience also caused differences in prescription compatibility and drug dosage, which makes difficult guarantee for the safety and effectiveness of drug treatment. So, this paper is based on literatures about the traditional Chinese medicine in treatment of cervical vertigo in the past 5 years, by using association rules algorithm, complex system entropy clustering and other non supervised data mining methods. Analysis was made for the use of various drugs in the frequency, the association rules, the core drug combination and the new prescriptions. The traditional Chinese medicine inheritance support system(TCMISS) was utilized to analyze 154 drugs from the 101 prescriptions, determine the use frequency of drugs, analyze the characteristics and the compatibility of core drugs, and dig out 14 core drug combinations and 7 new prescription combinations. The prescription and medication regularities illustrates the drugs for treatment of cervical vertigo, including those for flating liver and suppressing yang, invigorating the circulation of blood to remove blood stasis, reducing water and permeating dampness, increasing qi and activating blood, and nourishing the liver and kidney mainly. Treatment rules are nourishing liver and kidney, invigorating the circulation of blood stasis Tongqiao, reducing phlegm and dampness, flating liver and suppressing yan, dredging collaterals, supplementing qi and nourishing blood. This study aims to summarize frequently used single herbs for vertebral artery type cervical spondylosis, combinations of frequently used herbs and dosage of frequently used herbs with significant efficacy, define the current prescription and medication regularities for treating cervical vertigo and give guidances for clinical mediation.

9.
Artigo em Chinês | WPRIM | ID: wpr-464953

RESUMO

BACKGROUND:Extensive studies have shown that transcutaneous electrical nerve stimulation (TENS) plays a positive role in relieving the pain caused by a variety of diseases. However, its exact effect to manage pain in patients with knee osteoarthritis is stil controversial. TENS is classified into h-TENS and l-TENS, but currently its respective role in relieving the pain caused by knee osteoarthritis is not clear yet. OBJECTIVE: To compare the efficacy of h-TENS and l-TENS on pain relieving among patients with knee osteoarthritis. METHODS: A computer-based search was performed on PubMed, Embase and Cochrane database for randomized controled trials on TENS for the treatment of knee pain in patients with knee osteoarthritis which were reported before February 2014. Methodology quality of the trials was criticaly assessed and relative data were extracted. These studies were selected independently by two reviewers according to the inclusion and exclusion criteria. Meta-analysis was performed using Revman5.2 software. RESULTS AND CONCLUSION:A total of 10 randomized controled trials involving 519 cases were included. The results of meta-analysis showed that the h-TENS group had significant effects on pain relief in knee osteoarthritis patients in comparison to the control group [MD=-0.56, 95%CI(-0.98,-0.15),P=0.008]; the l-TENS group had no significant effect on pain relief in knee osteoarthritis patients in comparison to the control group [MD=-0.13, 95%CI(-1.63, 1.38),P=0.87]; the h-TENS group had significant effect on pain relief in knee osteoarthritis patients in comparison to the l-TENS group [MD=-0.85, 95%CI(-1.32, -0.37), P=0.000 5]. These findings indicate that h-TENS performs something positive to reduce pain in knee osteoarthritis patients that l-TENS cannot do. Owing to the limitations of this study, further work is needed to determine the role of TENS in pain management among patients with knee osteoarthritis.

10.
Artigo em Chinês | WPRIM | ID: wpr-454611

RESUMO

BACKGROUND:Symptomatic treatment is the main management strategy for patients with knee osteoarthritis at early metaphase. Some previous studies have demonstrated that neuromuscular electrical stimulation can al eviate pain in knee osteoarthritis patients. To date, this effectiveness, however, stil remains controversial. OBJECTIVE:To assess the efficacy of neuromuscular electrical stimulation on pain in patients with knee osteoarthritis. METHODRandomized control ed trials concerning the efficacy of neuromuscular electrical stimulation in the treatment of knee osteoarthritis were identified from the Medline database conducted up to July 3, 2014. These studies were selected independently by two reviewers according to the inclusion and exclusion criteria. Meta-analysis was performed using Revman5.2 software. RESULTS AND CONCLUSION:A total of 5 randomized control ed trials consisting of 239 participants were included. The results of the meta-analysis indicate neuromuscular electrical stimulation has no significant impact on measure of pain in knee osteoarthritis patients in comparison to the blank control group [mean difference=-0.40, 95%confidence interval (-1.34-0.54), P=0.40]. Owing to the sample limitations of our study, it is hard for us to draw a conclusion that the application of neuromuscular electrical stimulation in managing pain in patients with knee osteoarthritis is of little significance. Further work based on large-sample and high-quality randomized control ed trials is needed to determine the role of neuromuscular electrical stimulation in pain in this population.

11.
Chinese Journal of Urology ; (12): 120-122, 2012.
Artigo em Chinês | WPRIM | ID: wpr-420785

RESUMO

Objective To evaluate the effect of transurethral feedback microwave thermotherapy with the ProstaLund CoreTherm Device (PLFT) in benign prostate hyperplasia ( BPH ) patients with high risk factors 24 months after treatment.MethodsSixty-two BPH cases with high risk factors of aged ≥ 80or complicating severe conditions of no less than one organ or system,were treated with PLFT under urethral local anesthesia.The average pre-treatment prostate volume,international prostate symptom score (IPSS),quality of life score (QOL) and maximal urinary flow (Qmax) were 62.03 ml,23.19,4.58 and 4.33 ml/s,respectively.The changes of prostate volume,IPSS,QOL and Qmax at 3 months,12 months and 24 months after treatment were analyzed.ResultsAll patients tolerated well of PLFT performed in common therapy room except lightly bleeding,minor infection and temporary incontinence.There was no severe surgical adverse event.After 3 months,the prostate volume reduced to 43.85 ml,IPSS decreased to 11.63,QOL decreased to 2.44,Qmax rose up to 11.44 ml/s; The average values were 45.10 ml,12.23,2.61 and 10.91ml/s at 12 months after treatment.The corresponding values were 45.80 ml,12.37,2.66 and 10.82 ml/s,respectively at 24 months after treatment.Compared with pre-treatment,all the parameters showed significant improvement ( P < 0.01 ).ConclusionsPLFT is one of the effective and safe treatment options for BPH patients with high risk factors.It can be safely used on day-surgery patients.The best effect appears at 3 months after treatment.

12.
Chinese Journal of Hepatology ; (12): 346-352, 2010.
Artigo em Chinês | WPRIM | ID: wpr-326362

RESUMO

<p><b>OBJECTIVE</b>To identify the active material of anti-hepatic fibrosis from Amydae Carapax.</p><p><b>METHODS</b>Membrane separation technology was adopted to screen active fraction in Amydae Carapax, and the active components were isolated from the active fraction using gel chromatography and high performance liquid chromatography. The purified active components in Amydae Carapax were further analyzed using 4700 series time-of-flight mass spectrometer.</p><p><b>RESULTS</b>Proteins and peptides of Amydae Carapax with molecular weight less than 6000 were proved to have biological activity. 8 components (Bj1-Bj8) were isolated from the active fraction. Bj4, Bj6 and Bj7 were screened as active components. Bj7 was further purified, resulting in 7 components (Bj701-Bj707). Bj704 and Bj707 showed significant biological activity. Mass spectrometry showed three molecular ion peaks with highest abundance, i.e. m/e 526, 542 and 572, i.e. m/e 526, 542 and 572, in Bj707 -A The amino acid sequences of above three peptide compounds were NDDY (Asn-Asp-Asp-Tyr), NPNPT (Asn-Pro-Asn-Pro-Thr), and HGRFG (His-Gly-Arg-Phe-Gly), respectively. And M572 was the most abandunt components.</p><p><b>CONCLUSION</b>Three active peptide compounds of anti-hepatic fibrosis of Amydae Carapax were identified.</p>


Assuntos
Animais , Humanos , Linhagem Celular , Cirrose Hepática , Medicina Tradicional Chinesa , Extratos de Tecidos , Farmacologia
13.
Chinese Journal of Urology ; (12): 113-115, 2010.
Artigo em Chinês | WPRIM | ID: wpr-390955

RESUMO

Objective To evaluate the effect of transurethral feedback microwave thermotherapy with the ProstaLund CoreTherm Device(PLFT) in high risk patients with benign prostate hyperplasia (BPH). Methods Sixty-six high risk patients diagnosed with BPH, including aged ≥80 in 32 pa-tients, hypertension in 31 patients, diabetes in 5 patients, heart failure in 8 patients, chronic obstruc-tive pulmonary disease in 8 patients, cerebral infarction in 11 patients, fracture, amputation or joint stiffness unsuitable for lithotomy position in 3 patients, abnormal blood coagulation in 4 patients, pan-creatitis in 2 patients, cardiac arrhythmia in 6 patients and malignant tumor in 3 patients, were treated with PLFT using individual power at urethral local anesthesia, resulting in coagulation necrosis in 15%-30% of prostate tissue around urethra. Meanwhile, real-time monitoring the temperature of prostate and the tissue around it was used. All patients were evaluated by comparing volume of pros-tate, maximal urinary flow (Q_max), international prostate symptom score (IPSS) and quality of life questionnaire (QOL) in pre-treatment and three months after respectively. Results All of patients well tolerated PLFT. There was bleeding lightly, infection lightly and temporary incontinence. There was no severe surgical complication. After three months, the volume of prostate reduced from 62. 2 ml to 44.5 ml; IPSS decreased from 23. 4 to 11.7; QOL decreased from 4.5 to 2.4; Q_max rised from 4, 2 ml/s to 11.2 ml/s. All differences reached significance. Conclusion PLFT is one of effective and safe treatments for patients with BPH especial BPH complicating with severe conditions.

14.
Artigo em Chinês | WPRIM | ID: wpr-567846

RESUMO

Objective To investigate the effect of calcium/calmodulin-dependent protein kinaseⅡ (CaMK[KG-*6]Ⅱ) inhibitor,KN93 on calcium overloading of atrial muscle cells in neonatal rats and detect the expression of CaMK[KG-*6]Ⅱ. Methods The atrial muscle cells from neonatal rats were primarily cultured for 96 h and then divided into 6 group,control,calcium overloading group,KN93 group (0.5 ?mol/L),low-,moderate-and high-dose of KN93+ calcium overloading group. A model of calcium overloading for atrial muscle cells was established by using calcium ionophore (ionomycin,1.0 ?mol/L). For the later 3 groups,KN93 at doses of 0.25,0.5 and 1.0 ?mol/L was added into the culture medium for 30 min followed by 1.0 ?mol/L ionomycin treatment for another 30 min. The identification of ?-actin was performed by immunofluorescence staining. In the present of Fluo-3/AM (an indicator of calcium),intracellular calcium and the expression of CaMK[KG-*6]Ⅱ were detected under the intervention of KN93 with laser cofocal microscopy and Western blotting respectively. Results More than 90% of cultured cells were positive to ?-actin antibody. Compared with the control group,the fluorescence intensity of intracellular Ca2+ was increased significantly by ionomycin (660.16?108.47 vs 376.12?57.57,P

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