Study on relationship between autophagy and drug resistance of leukemia K562/ADM cells / 中国药理学通报

Aim To set up leukemic K562/ADM cells with stable tolerance to 15 fimol • L_1 ADM induced in vitro by long-term and continuous stepwise increment of adriamycin (ADM) concentration, to observe the sensitivity to other chemotherapy drugs and the relationship between autophagy and drug resistance.Methods MTT assay was used to detect the sensitivity of cells to chemotherapy drugs.The morphological changes of autophagy were observed by transmission electron microscope and fluorescence microscopy.Cell apoptosis analysis was performed using Annexin-V/PI double staining and flow cytometry ( FCM ).The expressions of autophagy and drug resistance associated proteins were tested by Western blot.Results K562/ADM cells were cross-resistance to the other chemotherapeu-tics besides adriamyciri, such as pirarubicin, daunoru- bicin, 5-flurouracil, vincristine but not arsenic triox- ide.The number of autophagosomes, the fluorescence intensity of monodansylcadaverine (MDC) and the expression of LC3-H ,Beclin-l in K562/ADM cells were significantly higher than those in K562 cells.The inhibition of autophagy by 3-MA significantly increased the sensitivity of K562/ADM cells to ADM, and 3-MA also effectively inhibited the expressions of drug resistance related proteins P-gp, MRP1 and BCRP in K562/ADM cells.Conclusions The K562/ADM cells resistant to adriamycin occur multidrug resistance, and the drug resistanceis closely related to the level of autophagy.

Kaempferol in reversing drug resistance of chronic myelogenous leukemia K562/ADM cells and its related mechanism / 白血病·淋巴瘤

Objective@#To investigate the drug resistance of kaempferol reversed adriamycin (ADM)-resistant K562/ADM cells in chronic myelogenous leukemia (CML) and its related mechanism.@*Methods@#Methyl thiazolyl tetrazolium (MTT) method was used to detect the toxicity of ADM on K562 and K562/ADM cells for 24 h. The half inhibitory concentration (IC50) of ADM and the drug resistance multiple for 24 h were calculated. MTT method was used to detect the toxicity of kaempferol on K562/ADM cells for 24 h. The 5% inhibitory concentration (IC5) and 10% inhibitory concentration (IC10) of kaempferol for 24 h were calculated to determine the concentration of kaempferol in the subsequent experiments. And the cells untreated by the kaempferol were selected as the control group. The cell inhibition after the treatment of ADM for 24 h of the blank control group and kaempferol intervention group was detected by using MTT method. And then the cell inhibition for 24 h and ADM IC50 for 24 h in the above groups were calculated. The ratio of IC50 in the blank control group and kaempferol group was the reversal drug resistance multiple of kaempferol. The fluorescence intensity of ADM in K562/ADM cells treated by kaempferol was detected by using flow cytometry. Western blotting was used to detect the expressions of P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1), phosphorylated p38 (p-p38), and total p38 (t-p38) protein in K562/ADM cells after the treatment of kaempferol, the specific inhibitor of p38-MAPK signaling pathway SB202190, and the combination of kaempferol and SB202190.@*Results@#After the treatment of ADM for 24 h, the IC50 value of K562 and K562/ADM cells was (0.9±0.6), (28.1 ±3.5) μg/ml, respectively. The drug resistance multiple of K562/ADM cells on the treatment of ADM for 24 h was 31.16 compared with the K562 cells. MTT method showed that kaempferol inhibited the proliferation of K562/ADM cells in a dose-dependent manner. According to the IC5 and IC10, 0.5 μmol/L and 1.0 μmol/L kaempferol were determined to do the subsequent experiments. After the combined interaction of kaempferol and ADM for 24 h, the ADM IC50 of K562/ADM cells in the blank control group, 0.5 μmol/L kaempferol group and 1.0 μmol/L kaempferol group was (33.7±5.7), (21.4±0.6), (15.9±1.8) μg/ml, respectively (F = 30.85, P < 0.05), and there was a statistical difference of pairwise comparison (both P < 0.05). The reversal drug resistance multiple of K562/ADM cells for 24 h in 0.5 μmol/L kaempferol group and 1.0 μmol/L kaempferol group was 1.58 and 2.12, respectively. Flow cytometry results showed that the mean fluorescence intensity (MFI) of ADM in the blank control group, 0.5 μmol/L kaempferol group and 1.0 μmol/L kaempferol group was 138.4±8.9, 154.3±2.2, 165.7±4.8, respectively, and the difference was statistically significant (F = 161.48, P < 0.05). Compared with the blank control group, after treatment of K562/ADM cells with 0.5 μmol/L and 1.0 μmol/L kaempferol for 24 h, the relative expressions of P-gp, MRP1 and p-p38 protein were decreased in K562/ADM cells (all P < 0.05), but there was no statistical difference in the expression of t-p38 protein (P > 0.05); SB202190 could reduce the relative expressions of P-gp, MRP1 and p-p38 protein (all P < 0.05); after the treatment of SB202190 combined with different concentration of kaempferol, the relative expressions of P-gp, MRP1 and p-p38 protein in K562/ADM cells did not decrease (P > 0.05).@*Conclusions@#Kaempferol can decrease the relative expressions of P-gp and MRP1 in K562/ADM cells by inhibiting p38-MAPK pathway, so as to increase the concentrations of ADM and to reverse the drug resistance of K562/ADM cells.

Inhibitory effect of PI3Kδ inhibitor idelalisib on proliferation of human myeloid leukemia cells and the reversal effect on drug resistance to adriamycin / 中南大学学报(医学版)

OBJECTIVES@#To investigate the effect of adriamycin (ADM), idelalisib or ADM and their combination on cell proliferation and intracellular concentration of ADM, and to explore the reversal effect of idelalisib on drug resistance to ADM.@*METHODS@#The K562 and K562/ADM cells were respectively treated with ADM and idelalisib at different concentrations. The 50% inhibitory concentration (IC@*RESULTS@#The cell survival rates were significantly decreased in a dose-dependent manner when the cells were treated with different doses of ADM (0.001-10.000 mg/L ). The IC@*CONCLUSIONS@#Idelalisib exerts effect on inhibition of the proliferation in myeloid leukemia K562 and K562/ADM cells, which may partially reverse the drug resistance of K562/ADM cells to ADM. The mechanisms for the effect of idelalisib may be related to increasing the accumulation of ADM and inducing the cell apoptosis in the K562 and K562/ADM cells.

Study on the Mechanism of Periplaneta americana Extract Reversing Multi-drug Resistance of Human Hepato- cellular Carcinoma HepG 2/ADM Cells / 中国药房

OBJECTIVE：To study the mechanism of Periplaneta americana extract degreasing cream and CⅡ-3（shorted for “degreasing cream ”and“CⅡ-3”）reversing the multi-drug resistance of human HepG 2/ADM cells. METHODS ：MTT assay was used to investigate the toxicity effects of different concentrations of sorafenib （positive control ），degreasing cream and C Ⅱ-3 on HepG2/ADM cells ，then IC 20 was calculated. The experiment was divided into sensitivity drug ，drug-resistance group ，sorafenib group，degreasing cream group and C Ⅱ-3 group. HepG 2 cells were included in sensitivity group ，and HepG 2/ADM cells were included in the latter 4 groups. Sensitivity group and drug-resistance group were treated with routine medium ，and other 3 groups were treated with relevant medicine （IC20 as drug concentration ）. The content of ADM in HepG 2/ADM cells was determined by Laser scanning confocal microscopy. The expression of apoptosis-related protein as Bcl- 2 and Cleaved-Caspase- 9 p37 were detected by Western blotting assay. RT-qPCR and immunocytochemistry were adopted to detect mRNA and protein expressions that related to multidrug resistance [P-gp （expression produce of MDR1 gene），LRP，BCRP] and that related to enzyme-mediated multidrug resistance pathway （GST-π and Topo Ⅱ）. RESULTS ：The IC 20 of degreasing cream ，CⅡ-3 and sorafenib were （2.40±0.16）， （200.44±27.52），（18.00±1.82）μg/mL，respectively. Compared with sensitivity group ，the protein expressions of Bcl- 2，P-gp， LRP，BCRP and Topo Ⅱ，the mRNA expressions of MDR 1， LRP，BCRP and GST-π were increased significantly in drug resistance group （P＜0.05 or P＜0.01）. Compared with @qq.com drug-resistance group ，the mRNA and protein expression of MDR1 mRNA and LRP ，BCRP，GST-π were significantly decreased in degreasing cream group and C Ⅱ-3 group（P＜ 0.05 or P＜0.01）；the protein expression of Bcl- 2 and the mRNA expression of Topo Ⅱ were significantly decreased （P＜0.01）， while the protein expression level of Cleaved-Caspase- 9 p37 was significantly increased in C Ⅱ -3 group （P＜0.05）. CONCLUSIONS：Degreasing cream and C Ⅱ-3 can reverse multidrug resistance of HepG 2/ADM cells by reducing drug efflux ， promoting cell apoptosis ，reducing the mRNA and protein expression of multi-drug resistance gene as well as gene in enzyme-mediated multi-drug resistance pathway. The effect of C Ⅱ-3 is better than that of degreasing cream.

Application status of commonly used patches in breast reconstruction / 中国肿瘤临床

Breast reconstruction is an essential part in the comprehensive management of breast cancer. The clinical application of patches (e.g., acellular dermal matrix, ADM) is the most impactful innovation in implant breast reconstruction in recent years. The wide application of patches in implant breast reconstruction promotes the development of immediate prosthetic reconstruction, im-proves the aesthetic outcomes of reconstructed breasts, and avoids additional donor tissue damage caused by autologous flap breast reconstruction. At present, patches used in breast reconstruction are mainly ADMs, bovine pericardial patches, and TiLOOP, which are widely used because of their good histocompatibility and tissue defect repair ability. This article reviews the applications and research statuses of these patches.

Enhancing effect of chidamide on the sensibility of human chronic myeloid leukemia K562/ADM cells to daunorubicin / 解剖学报

Objective: To investigate whether chidamide (CDM) could influence the sensibility of human chronic myeloid leukemia K562/ADM cells to daunorubicin (DNR) and its possible mechanism. Methods: The K562 and K562/ADM cells were cultured in vitro and treated with CDM and(or) DNR for 48 hous, and then the cell viability was measured by cell counting kit-8(CCK-8) assay. The proliferation, cell cycle and apoptosis were analyzed by flow cytometry. Western blotting was performed to measure the protein levels of histon 2AX (H2AX), γH2AX (Serl39), ataxia telangiectasia mutated gene (ATM), p-ATM (Serl981), breast cancer susceptibility protein 1(BRCAl), and p-BRCAl (Serl524). Results: DNR remarkably inhibited the cell activity of K562/ADM cells in dose-dependent manner with a half maximal inhibitory concentration(IC50) value of 11.76 μmol/L, and the resistant factor was 18.09. Co-treatment with CMD and DNR produced a synergistic effect confidence interval(GI) (CI<1) with a reversal fold of 8.11. DNR remarkably inhibited proliferation (P<0.05), induced G2/M phase arrest and apoptosis (P<0.05), these effects were enhanced under non-toxic concentration of CMD (P<0.05). K562/ADM cells had a significantly higher protein levels of ATM and BRCA1 than K562 cells (P<0.05). DNR significantly up-regulated the protein levels of γH2AX, p-ATM and p-BRCAl (P<0.05), and the protein level of γH2AX appeared higher in the combination group compared to DNR alone (P<0.05); however, the co-treatment with CMD and DNR induced a decreased expression of p-ATM and p-BRCAl than the DNR alone (P< 0.05). Conclusion: CDM may enhance the sensibility of K562/ADM cells to DNR by up-regulating the protein level of γH2AX, and down-regulating the protein levels of p-ATM and p-BRCAl.

Effects of combined oral doxycycline and topical cyclosporine treatment on ocular signs, symptoms, and tear film parameters in rosacea patients / Efeitos da doxiciclina oral combinada e tratamento tópico com ciclosporina nos sinais e sintomas oculares e em parâmetros do filme lacrimal em pacientes com rosácea ocular

ABSTRACT Purpose: This study reports the effects of combined use of oral doxycycline and topical cyclosporine on ocular signs, symptoms, and tear film parameters in rosacea patients. Methods: Fifty-four right eyes of 54 patients were included in this study. All patients underwent full ophthalmologic examination-including best corrected visual acuity measurement, slit-lamp anterior segment and fundus examination, tear film break-up time, and Schirmer test-before treatment and six months post-treatment. Patients were divided into two treatment groups. The first group was treated with oral doxycycline 100 mg twice daily for the first month and once daily for the following two months. The second group received topical 0.05% cyclosporine emulsion drops twice daily for six months in addition to the oral doxycycline treatment regimen. All patients received preservati ve-free artificial tear drops, warm compress, eyelash cleaning, and topical corticosteroid drops three times daily for one month. Results: A significant improvement in ocular signs and symptoms was recorded for all patients in groups 1 and 2 after treatment. There was not a significant difference in terms of itching, burning, meibomian gland inspissation, corneal neovascularization, and conjunctival hyperemia score changes between groups 1 and 2. The increases in Schirmer test and break-up time scores were significantly higher in group 2 than in group 1. Conclusions: Our results support the finding that topical cyclosporine in addition to the standard regimen improves tear function, as shown by Schirmer test and break-up time scores, in ocular rosacea patients.

RESUMO Objetivo: Este estudo relata os efeitos do uso combinado de doxiciclina oral e ciclosporina tópica sobre sinais e sintomas oculares e sobre parâmetros do filme lacrimal em pacientes com rosácea. Métodos: Cinquenta e quatro olhos direitos de 54 pacientes foram incluídos no estudo. Todos os pacientes foram submetidos a exame oftalmológico completo - incluindo a melhor medida da acuidade visual corrigida, segmento anterior em lâmpada de fenda e exame de fundo de olho, tempo de ruptura do filme lacrimal e teste de Schirmer - antes do tratamento e após seis meses de tratamento. O primeiro grupo foi tratado com doxiciclina oral 100 mg duas vezes ao dia no primeiro mês e uma vez ao dia nos dois meses seguintes. O segundo grupo recebeu gotas tópicas de emulsão de ciclosporina a 0,05% duas vezes ao dia por seis meses, além do tratamento com doxiciclina por via oral. Todos os pacientes receberam gotas de lágrima artificial sem conservantes, compressas mormas, limpeza de cílios e gotas de corticosteróide tópico três vezes ao dia durante um mês. Resultados: Uma melhora significativa nos sinais e sintomas oculares foi registrada para todos os pacientes do grupo 1 e 2 após o tratamento. Não houve diferença significativa em termos de prurido, queimação, inspeção da glândula meibomiana, neovascularização da cór nea e alterações na pontuação da hiperemia conjuntival entre os grupos 1 e 2. O teste de Schirmer e o aumento do tempo de ruptura no grupo 2 foram significativamente maiores do que no grupo 1. Conclusões: Os autores concluíram que os resultados apoiam a descoberta de que a ciclosporina tópica, além do tratamento padrão, melhora a função lacrimal como demonstrado pelo teste de Schirmer e o tempo de ruptura em pacientes com rosácea ocular.

Predictive Factors In Successful Completion Of Laparoscopic Cholecystectomy In Acute Cholecystitis.

Background: Laparoscopic Cholecystectomy in acute Cholecystitis is the established treatment of choice for the management of acute Cholecystitis. The conversion rate and morbidity is higher in Laparoscopic Cholecystectomy done in acute Cholecystitis. Aims and Objectives: To identify certain preoperative criteria for the selection of patients in acute Cholecystitis that can undergo laparoscopic Cholecystectomy, so that there is decrease in the conversion rate to open procedure and decrease in the complications. Methods: A Prospective interventional study was conducted in the Department of General Surgery, Moolchand Medcity, New Delhi, on 50 patients with acute Cholecystitis from September 2009 to September 2010. Data was analyzed using standard statistical software SPSS. Chi-square test was used for statistical analysis of qualitative data. Results: There were 4 males and 46 female patients with a mean age of 37. 74 years (range 18 to 65 years). There was no significant variation in intraoperative severity (IOS) pattern of acute Cholecystitis across the various age groups (p=0. 532). Males had significantly higher IOS grades of acute Cholecystitis as (p=0. 000). Among patients with varying IOS of acute Cholecystitis, there was significant difference in mean values of duration of symptoms before surgery (p=0.006), TLC at admission (adm.) (p=0.037) and at 24 hours of adm. (p=0.016) and serum AMS at adm. (p=0.005). The difference in mean serum ALP at adm. (p=0. 0171), at 24 hours of adm. (p=0. 137) and at 48 hours of adm. (p=0. 151), and mean values of TLC (p=0. 052) at 48 hours of adm. were insignificant across the various IOS grades of acute cholecystitis. There was no significant variation in histopathological severity (HPS) pattern of acute cholecystitis across the various age groups (p=0.826). Males had significantly higher HPS grades of acute Cholecystitis compared to females (p=0.042). Among patients with varying HPS of acute cholecystitis, there was no significant difference in mean duration of symptoms before surgery (p=0. 065). The difference in mean values of TLC at adm. (p=0.001), at 24 hours of adm. (p=0.001) and 48 hours of adm. (p=0.003); serum ALP at adm. (p=0.001), at 24 hours of adm.(p=0.001) and at 48 hours of adm.(p=0.022) was very significant across the various HPS grades of acute cholecystitis. The IOS (p=0.035) and HPS (p=0.032) of acute cholecystitis was significantly less with successful early lap Chole than with failed procedure. There was no significant difference in success versus failure rates of early Lap Chole across the age distribution of the patients (p=0. 153); mean values of duration of symptoms before surgery (p=0.971); TLC at adm. (p=0.422), at 24 hours of adm. (p=0.990) and at 48 hours of adm. (p=0. 478); serum ALP at adm. (p=0.113), at 24 hours of adm. (p=0.135) and at 48 hours of adm.(p= 0. 238). Male patients had significantly higher failure rate of early Lap Chole (p=0.022). Patients with failed early Lap Chole had a significantly longer mean duration of surgery (p=0. 032) and postoperative hospital stay (p=0.028) than those undergoing a successful procedure. Conclusion: Acute Cholecystitis is much more common in females. Severity of inflammation (intraoperative/ histopathological) in acute Cholecystitis is associated with higher failure/conversion rate of early laparoscopic Cholecystectomy because of dense pericholecystic adhesions, and has longer duration of surgery and postoperative hospital stay. This has a positive relation with male sex and TLC(IOS and TLC at 48 hrs of adm. , however had insignificant relation in our study ), while as no relation with age (at least up to 65 years) and mean duration of symptoms before surgery (if surgery is done within 1.5-4 days of onset of symptoms).

Study on the drug-resistant reversal effects of ginsenoside Rh2 in human hepatocellular carcinoma HepG2/ADM cells and its mechanism / 医学研究生学报

Objective Ginsenoside Rh2 can inhibit the proliferation of a variety of malignant tumor cells.However, little research has been done on the sensitivity of Rh2 in human hepatocellular carcinoma HepG2/ADM cells with multidrug resistance(MDR).This study aimed to explore the reversing effects of ginsenoside Rh2 on the MDR of human hepatocellular carcinoma HepG2/ADM cells and its potential mechanism.Methods MTT assay was applied to detect the effect of Rh2(0-250 μg/mL) on the viability of HepG2/ADM cells and screen out the optimum drug-resistant reversal concentration of Rh2.Cells were divided into 3 groups: HepG2/ADM group (without any medicine treatment), ADM group(ADM treatment for 48 h), ADM+40 μg/mL Rh2 group (pretreatment of 40μg/mL Rh2 for 30 min followed by ADM treatment for 48 h).Flow cytometry was applied to detect the effect of Rh2 on the fluorescence intensity of cellular Rh-123.RT-PCR was used to measure the expression of MDR1 gene.Western blot was used to detect the protein levels of P-gp, Bax, Bcl-2 and cleaved caspase-3.Results 40 μg/mL ginsenoside Rh2 significantly reversed the MDR of HepG2/ADM cells by a 2.55-to-3.70-fold increase in sensitivity.Furthermore, compared with ADM group, the efflux of Rh-123 in HepG2/ADM cells were remarkably inhibited by Rh2 in ADM+40 μg/mL Rh2 group (65.83±1.78 vs 78.21±1.26, P<0.01), along with the down-regulated expressions of MDR1 (0.48±0.02 vs 0.86±0.05, P<0.05), P-gp(0.97±0.04 vs 1.91±0.03,P<0.01), Bcl-2(1.25±0.05 vs 1.86±0.03, P<0.05) and the up-regulated protein level of Bax (1.76±0.04 vs 1.25±0.02,P<0.05) and cleved caspase-3(0.42±0.04 vs 38.26±5.45,P<0.05).Conclusion Ginsenoside Rh2 can effectively reverse the MDR of HepG2/ADM cells, and the potential mechanism is related to the decreased expressions of MDR1 and P-gp, the increasing drug concentration inside the cells and the Bax/Bcl-2 signaling pathway.

Influence of Hiwigene targeting silence in chemotherapy sensitivity of breast cancer MCF-7/ADM cells / 吉林大学学报(医学版)

Objective:To explore the changes of chemotherapy sensitivity of breast cancer MCF-7/ADM cells after Hiwi gene targeting silence, and to illuminate the role of Hiwi gene in resistant mechanism of breast cancer MCF-7/ADM cells.Methods:The MCF-7/ADM cells were transfected with Hiwi control,siRNA1 Hiwi gene and siRNA2 Hiwi gene as Hiwi control group, Hiwi siRNA1 group and Hiwi siRNA2 group.The expression levels of Hiwi gene mRNA and protein in breast cancer MCF-7/ADM cells were detected by Real-time PCR and Western blotting methods to judge the transfection rates in various groups.After transfection,the breast cancer MCF-7/ADM cells in various groups were treated with the different concentrations(0,0.1,0.5 and 1.0 mg·L-1) of adriamycin, and the cell resistant sensitivities were detected by MTT method.Results:Compared with control group(0 mg·L-1 adriamycin), the expression levels of Hiwi gene mRNA and protein inMCF-7/ADM cells in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased (P<0.01).Compared with control group, the survival rates of Cells in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased after treated with different concentrations of adriamycin,and the survival rates in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased(P<0.01);they were (48.15±6.28)% and (41.73±6.17)% when the concentration of adriamycin was 1 mg·L-1,and the sensitivities to the adriamycinwere obviously enhanced(P<0.01).Conclusion:The target silencing Hiwi genes in MCF-7/ADM cells is efficient.The sensitivity of MCF-7/ADM cells to adriamycin restores after Hiwi gene silencing.

An Electrode Configuration for Recording Muscle Motor Evoked Potentials in the Upper Extremities during Intraoperative Neurophysiological Monitoring

OBJECTIVE: The main aim of the present study is to examine the electrode configurations used to record the muscle motor evoked potential (mMEP) in the upper extremities during surgery with the goal of producing a high and stable mMEP signal, in particular among the abductor pollicis brevis (APB), abductor digiti minimi (ADM), and across the APB-ADM muscles, which have been widely used for the mMEP in the upper extremities. METHODS: Thirty right-handed patients were recruited in this prospective study. No patients showed any adverse events in their mMEP signals of the upper extremities during surgery. The mMEPs were recorded independently from the signals for the APB and ADM and for those across the APB-ADM. RESULTS: The mMEP amplitude from across the APB-ADM was statistically higher than those recorded from the APB and ADM muscles. Moreover, the coefficient of variation of the mMEP amplitude from across the APB-ADM was smaller than those of mMEP amplitude recorded from the APB and ADM muscles. CONCLUSION: The mMEP from across the APB-ADM muscles showed a high yield with high stability compared to those in each case from the APB and ADM muscles. The configuration across the APB-ADM muscles would be best for mMEP recordings from the upper extremities for intraoperative neurophysiological monitoring purposes.

Reverse Effect of Interferon-α Combined with All-Trans Retinoic Acid on Multidrug Resistance of Human Leukemic Cell Line K562/ADM / 中国药学杂志

OBJECTIVE: To investigate the effects of interferon-α(IFN-α) and all-trans retinoic acid(ATRA) on multidrug resistance reversal effect and mechanism of human leukemia K562/ADM cells. METHODS: The cytotoxicity and reversal times of IFN-α and ATRA were detected by CCK-8 method. Apoptosis rate and cell cycle were detected by flow cytometry. PI3K, Akt and Bad mRNA were detected by RT-PCR method. Western blot method was used to detect the expression of PI3K, AKt, P-AKt and Bad protein.RESULTS: The drug resistance of K562/ADM cells to adriamycin(ADM) was 54 times. ADM, respectively, with IFN-α, ATRA or combined application, the drug resistance of K562/ADM cells to ADM was 1.24, 2.34 and 8.14, respectively. The apoptosis rate of K562/ADM cells was significantly increased by using ADM 4 mg·L-1alone or in combination with IFN-α 2.5×106 U·L-1, ATRA 7.5 μmol·L-1, and the cell cycle was blocked in G0/G1 phase. PI3K mRNA and protein expression were significantly lowered, Akt mRNA and protein has no obvious change, Bad mRNA and protein expression are raised, phosphorylated Akt protein expression decreased, the expression is more obvious when the two drug combination. CONCLUSION: IFN-α and ATRA can reverse the multidrug resistance of K562/ADM cells, its mechanism may be the inhibition of the PI3K/Akt pathway.

Study on effect of curcumin in inhibiting proliferation of multidrug resistant liver cancer HepG2/ADM cells and its mechanism / 重庆医学

Objective To observe the inhibitory effect of curcumin on the proliferation of multidrug resistance liver cancer line HepG2/ADM cells and to explore its mechanisms.Methods HepG2/ADM cells were prepared and cultured in vitro,and treated by different concentrations (5,10,20,40 μmol/L) of curcumin for 24,48,72 h respectively.The effect of curcumin on proliferation of HepG2/ADM cells was measured by CCK-8 reagent;the concentration of intracellular rhodamine-123 (Rh-123) and adriamycin (ADM) were determined by flow cytometry;the level change of intracellular mdr-1 mRNA in each group was determined by RT-PCR,the P-gp protein level was detected by Western blot.Results Compared with the blank control and DMSO group,curucmin had more obvious inhibitory effect on HepG2/ADM cells proliferation (P＜0.05),and could more remarkably inhibit the intracellular Rh-123 excretion(P＜0.05).The RT-PCR and Western blot results showed that curcumm more significantly decrease the mdr-1 mRNA and P-gp protein levels in dose-time dependent manner (P＜0.05).Conclusion Curcumin could significantly inhibit the proliferation of multidrug-resistant HepG2/ADM cells,and its mechanism may be related with inhibiting mdr-1 gene expression and its encoded P-gp protein level,which are closely related with MDR.

Changes of plasma adrenomedullin in children with congenital heart disease combined with heart failure and their clinical implication / 实用医学杂志

Objective To explore the changes of plasma level of adrenomedullin(ADM)in congenital heart disease patients with heart failure,analyze the relationship between ADM and the severity of heart failure, assess the value of plasma ADM in the diagnosis of CHF and evaluate the cardiac function in pediatric patients with congenital heart disease. Methods 46 patients with CHD were enrolled according to the modified Ross Score. They were divided into three groups:mild CHF group(n = 16)and moderate CHF group(n = 18)and severe CHF group(n = 12). 25 matched normal children were enrolled as controls. Plasma ADM was measured with radioimmunoassay(RIA)and the contents of NT-proBNP were determined among all the patients. LVEF and E/A in all of them were also detected. The relationship between plasma ADM level and modified Ross Score and echocardiographic cardiac functional indexes was analyzed. Results Plasma ADM was positively correlated with modified Ross Score(r = 0.65,P < 0.01). Plasma ADM concentration in the severe CHF group was significantly higher than that of mild and moderate CHF group(P < 0.01),and plasma ADM concentration in the mild CHF group was higher than that in the control group(P<0.05).Plasma ADM was negatively correlated with LVEF and E/A(r =-1.05,P < 0.01). Conclusion Plasma ADM level could be used to assess cardiac function and diag-nose CHF with CHD.

Relationship between serum levels of ADMA ,CRP and atherosclerosis ,cardiovascular events in patients with type H hypertension / 国际检验医学杂志

Objective To investigate the relationship between serum levels of ADMA ,CRP and atherosclerosis ,cardiovascular e-vents in patients with type H hypertension .Methods Totally 80 patients with type H hypertension from August 2012 to August 2015 in our hospital were selected ,80 cases of non H type essential hypertension patiens were recruited as control group ,enzyme linked immunosorbent assay (ELISA) was used to detect serum levels of ADMA and CRP ,and carotid atherosclerosis was evalua-ted by ultrasonography .All patients were followed up regularly to record the occurrence of cardiovascular events ,and to investigate the relationship between ADMA and CRP levels and atherosclerosis and cardiovascular events .Results The levels of ADMA ,CRP and the rate of atherosclerosis in patients with type H hypertension were higher than those in the control group ,the difference was statistically significant (P<0 .05) .There were 37 cases of atherosclerosis in patients with type H hypertension ,and the levels of ADMA and CRP in patients with atherosclerosis were higher than those in non atherosclerosis group ,and the difference was statis-tically significant (P<0 .05) .Logistic regression analysis indicated that mean arterial pressure and ADMA levels were independent risk factors for cardiovascular events in patients (P<0 .05) .Conclusion Serum levels of ADMA are closely related to atherosclero-sis ,and are independent risk factors for cardiovascular events in patients with type H hypertension .

Relationship between serum levels of ADMA ,CRP and atherosclerosis ,cardiovascular events in patients with type H hypertension / 国际检验医学杂志

Objective To investigate the relationship between serum levels of ADMA ,CRP and atherosclerosis ,cardiovascular e-vents in patients with type H hypertension .Methods Totally 80 patients with type H hypertension from August 2012 to August 2015 in our hospital were selected ,80 cases of non H type essential hypertension patiens were recruited as control group ,enzyme linked immunosorbent assay (ELISA) was used to detect serum levels of ADMA and CRP ,and carotid atherosclerosis was evalua-ted by ultrasonography .All patients were followed up regularly to record the occurrence of cardiovascular events ,and to investigate the relationship between ADMA and CRP levels and atherosclerosis and cardiovascular events .Results The levels of ADMA ,CRP and the rate of atherosclerosis in patients with type H hypertension were higher than those in the control group ,the difference was statistically significant (P<0 .05) .There were 37 cases of atherosclerosis in patients with type H hypertension ,and the levels of ADMA and CRP in patients with atherosclerosis were higher than those in non atherosclerosis group ,and the difference was statis-tically significant (P<0 .05) .Logistic regression analysis indicated that mean arterial pressure and ADMA levels were independent risk factors for cardiovascular events in patients (P<0 .05) .Conclusion Serum levels of ADMA are closely related to atherosclero-sis ,and are independent risk factors for cardiovascular events in patients with type H hypertension .

Effect of Artesunate on Transferrin Receptor in K562/ADM Cells / 现代生物医学进展

Objective:To investigate the effect of Artesunate (Art) on the expression of transferrin receptor (TtR)in K562/ADM cells.Methods:The drug-resistant K562/ADM cells were cultured with 1000 ng/mL doxorubicin for two weeks followed by Artesunate treatment with different concentrations (12.5 μg/mL,25μg/mL and 50 μg/mL) or different time (12 h,24 h,36 h,and 48 h).The content of transferrin receptor in K562/ADM cells was determined by flow cytometry.The effect of Artesunate on the expression of transferrin receptor protein in K562/ADM cells was measured by Westem blot.Cell counting kit-8 (CCK-8) assay was used to evaluate inhibitory effect of Art combined with doxorubicin (ADM) in K562/ADM cells.The reversal index was defined as the IC50 of the experimental group divided by the IC50 of the control group in K562/ADM cells.Results:Art effectively decreased the content of transferrin receptor and the expression of transferrin receptor protein in K562/ADM cells in a dose-dependent manner.Moreover,Art also inhibited transferrin receptor protein expression in K562/ADM cells in a time-dependent manner.The different concentrations of Art(12.5 μg/mL,25μg/mL and 50 μg/mL) could induce reversal of drug-resistance with the reversal index being 1.38,2.12 and 2.95 times (P＜0.05).Art inhibited cell proliferation of K562/ADM cells,and the IC50 werel9.7 μmol/L.Conclusions:Art effectively down-regulated the transferrin receptor content as well as transferrin receptor protein expression in K562/ADM cells,which resulted in reversal of drug resistance of K562/ADM cells.Art also inhibited K562/ADM cells proliferation,which has great value in clinical treatment of leukemia.

The effects of different treatments of tooth extraction wound on alveolar bone healing / 实用口腔医学杂志

Bilateral mandibular first molars of 36 rabbits were extracted.The tooth extraction wounds were treated by:platelet-rich fibrin +acellular dermal matrix(group A),platelet-rich fibrin(group B),acellular dermal matrix (group C) and without treatment (group D,the control) (n =9) respectively.The measurments of alveolar bone height and width showed that there were no significant differences among groups at different times(P ＞ 0.05).Bone histomorphometry showed that at the 2th and 4th week,the best result was found in group A(P＜0.01).While,at the 8th week,the result of group A was still better than that of other 3 ones (P ＜ 0.01),but group B and C showed no significant difference(P ＞ 0.05).The combination of PRF and ADM shows the most significant effect.

Optimization of supercritical CO2 extraction for sauchinone and in vitro antitumor activity of Saururi Herba supercritical extract / 中成药

AIM To optimize the supercritical CO2 extraction of sauchinone and to evaluate the in vitro antitumor activity of Saururi Herba supercritical extract.METHODS With extraction pressure,extraction temperature,extraction time,entrainer (ethanol) concentration and entrainer amount as influencing factors,together with extraction rate of sauchinone as an evaluation index,orthogonal test was used for optimizing the extraction.Then MTT was applied to determining the extract's inhibitory effect on human multidrug-resistant hepatocellular carcinoma cell line (7721/Adm).RESULTS The optimal conditions were determined to be 30 MPa for extraction pressure,50 ℃ for extraction temperature,2 h for extraction time,95％ for ethanol concentration,and one time for ethanol amount,the average extraction rate of sauchinone was 0.173％.The obtained extract significantly inhibited the proliferation of 7721/Adm cells (IC50 =50.08 μg/mL),demonstrating a stronger activity than that of ethanol extract (ICs0 =150.59 μg/mL).CONCLUSION This stable and feasible method is appropriate for sauchinone extraction,and the supercritical CO2 extract from Saururi Herba shows a strong in vitro antitumor activity.

Expressions of ADM andβ-catenin in human osteosarcoma and their clinical significance / 西安交通大学学报(医学版)

ABSTRACT:Objective To study the expressions of adrenomedullin (ADM)andβ-catenin in human osteosarcoma of different differentiation grading and their clinical significance.Methods We chose 33 cases of human osteosarcoma with different pathological grading (14 cases of high differentiation grading and 1 9 cases of low grading)and 10 cases of human osteochondroma to detect the expressions of ADM protein,total (T)β-catenin and phosphorylated (P)β-catenin using immunohistochemical method. Results The expression of ADM was detected in the cytoplasm.In osteosarcomas of higher and lower differentiation grading,the positive rate of ADM was 57.1% (8/14)and 100% (1 9/1 9),respectively,with significant differences (P =0.0030.05).The expression of P-β-catenin was detected in the cytoplasm.In osteosarcomas of higher and lower differentiation grading,the positive rate of P-β-catenin was 35.7% (5/14)and 78.9% (1 5/1 9),with a significant difference (P =0.029 0.05 )or between T-β-catenin and P-β-catenin (P = 0.052 > 0.05 ).Conclusion The expressions of ADM and P-β-catenin proteins and the different distributions of expression of T-β-catenin protein in cytoplasm and/or nucleus are important indicators for judging the pathological grading of osteo-sarcoma.