OBJECTIVE: To investigate the effect of autophagy gene Beclin1 silencing on enhancing the drug sensitivity of imatinib-resistant human myeloid leukemia cell line K562/IMA. METHODS: Imatinib-resistant human myeloid leukemia cell line K562/IMA was constructed by the combination of the first dose of high-dose shock therapy and gradual increase of dose. Imatinib interfered with K562 and K562/IMA cell lines. Cell viability was detected by CCK-8. Apoptosis rate was detected by flow cytometry. Western-blot assay was used to determine the drug resistance level. The expression of Beclin1 in drug resistant strains was detected. Small interfering RNA (siRNA) transfected Beclin1 siRNA-Beclin1 was used to silence the Beclin1 gene of K562/IMA. Control group (control), siRNA negative control group (siRNA-NC) and Beclin1 siRNA group (siRNA-Beclin1) were set up. RT-QPCR and Western-blot were used to identify the expression level of Beclin1 protein and mRNA in each group after silencing. CCK-8 method was used to detect the sensitivity of each group of cells to imatinib, and the half inhibitory concentration IC50 was calculated. The apoptosis rate of each group was detected by flow cytometry. The expression levels of Beclin 1, Bcl-2, Bax, cleaved-caspase 3, caspase-3 and cytochrome C(cyto-C) were detected by Western-blot assay, and the metastatic and invasive ability of cells was detected by Transwell chamber. RESULTS: K562 / IMA cell lines were resistant to imatinib at 10 μmol•L-1, and the expression of Beclin1 was higher than that of K562 cell lines. The cell viability of K562/IMA was significantly higher than that of K562 cell lines in the gradient concentration range, while the apoptosis rate was lower than that of K562 cell lines. After siRNA silencing Beclin-1, the cell viability and apoptosis rate of siRNA-Beclin 1 group were significantly lower than those of control group, and the IC50 value of half inhibitory concentration was significantly lower than that of control group. The levels of Bax, cleaved-caspase 3, caspase-3 and cyto-C in the cells were significantly higher than those in the control group, while the levels of Bcl-2 were significantly lower than those in the control group. CONCLUSION: Beclin1 plays an important role in imatinib resistance in human myeloid leukemia, and autophagy may be involved in the drug resistance process. After silencing Beclin1, cell resistance level can be reduced and drug sensitivity improved.

OBJECTIVETo detect the DNA damage of the nurses occupationally exposed to anticancer drugs and to assess the exposure level using Comet assay.

METHODSSixteen nurses occupationally exposed to anticancer drugs were selected as exposure group, the average exposure period was 5.6 years, and the average exposure dose was to prepare 7.8 portions of anticancer drugs daily. Meanwhile, sixteen nurse students were selected as control group. The DNA migration of the peripheral lymphocytes of both groups was detected using comet assay.

RESULTSThe comet length was 46.27 microns in exposure group, which was significantly higher than that of control group (26.78 microns, P < 0.01). Also the percentage of long tailed nucleus (LTN) of exposure group was 64.83%, which was significantly higher than that of control group (4.87%, P < 0.01).

CONCLUSIONSThere was DNA damage in the nurses occupationally exposed to antineoplastic drugs.

Objective: To study the mechanism of interleukin (IL)-17 in mice with non-alcoholic fatty liver disease for promoting M1-type macrophage polarization to exacerbate liver inflammation, and to provide references for the mechanism of NAFLD occurrence and development. Methods: A mouse model of NAFLD was constructed by high-fat diet. Mice were divided into control group, model group, IL-17 group, and anti IL-17 group. Histopathological changes of the liver were observed by HE staining. The serum levels of ALT and AST in peripheral blood of mice was detected by chemical colorimetry. Macrophages labeled with F4/80-PE, CD11C-FITC was designated as M1-type macrophages, those labeled with F4/80-PE, and CD206-APC was designated as M2-type macrophages. The proportion of M1 and M2 macrophages infiltrated into the liver tissues of mice were measured by flow cytometry. CD168 expression level of liver tissues was detected using immunohistochemistry. Protein and mRNA levels of the marker molecules (iNOS, TNF-alpha and IL-6) of M1 macrophages were detected using ELISA and RT-Q PCR. Western blot was used to detect the protein expression of JAK-STAT signal pathway and the expression level of MCP-1. Data were analyzed using one-way ANOVA and t-test. Results: High-fat diet NAFLD mice model was successfully constructed. IL-17 had increased the proportion of M1 macrophages in mice liver tissues and decreased the proportion of M2 macrophages (P < 0.05). The proportion of M1 and M2 macrophages in the liver tissues of normal mice was 7.9% ± 1.1% and 19.2% ± 1.8%. The proportion of M1 and M2 macrophages in the model group was 17.3% ± 2.5% and 15.0% ± 2.1. The proportion of M1 macrophages (33.8% ± 4.2%) in IL-17 group was higher than model group, while the proportion of M2 macrophages (7.8% + 1.0%) in IL-17 group was lower than model group. Protein and mRNA marker levels of M1 macrophage (iNOS, IL-12, TNFα and IL-6) in liver tissues were significantly higher than model group, control group, and anti-IL-17 group (P < 0.05). The expression levels of JAK1, STAT1, MCP-1, and CD168 in mice liver tissues of IL-17 group had increased (P < 0.05). The levels of aspartate and alanine aminotransferases in peripheral blood of mice in IL-17 group were significantly higher than other three groups (P < 0.05). Conclusion IL-17 can promote M1-type macrophage polarization, and exacerbates the liver inflammatory response to accelerate the progression of NAFLD in mice.

Objective To investigate the role of nuclear factor(erythroid?derived 2)?like 2(Nrf2)in load?driven bone metabolism in Nrf2 knock?out(KO)mice. Methods The hybridized mice in the same brood were selected through PCR detection and were divided into two groups,i.e.,the Nrf2 knockout(KO)group and the wild?type(WT)group. Ulna of mice was loaded with 4 000 peak microstrains at 2 Hz for 3 consecutive days (120 cycles/day)as scheduled,the relative mineralizing surface(rMS/BS)and the relative bone formation rate(rBFR/BS)of ulna were measured for the two groups. Results Load?induced bone formation was suppressed in KO mice. Compared to the WT control,the relative bone formation rate was roughly 84%lower in KO mice(P<0.01). Conclusion The loss?of?function mutation of Nrf2 in bone diminishes load?driven bone formation.

Objective To investigate the significance of calcification in thyroid node for diagnosis of thyroid carcinoma.Method Retrospective analysis of 107 thyroid nodules' pre-operative ultrasonic and postoperative pathologic results.Results Total ultrasonic thyroid calcification ratio was 27.1%(29/107),which in benign samples was lower than Ihat in malignant samples(17.2%vs 70.0%,P＜0.01).Micro-calci-fication ratio in benign samples Was lower than thai:in malignanl samples(8.0%vs 50.0%,P＜0.01).Conclusion The ralio of thyroid carcinoma with calcification is higher,so the detection of thyroid carcinoma,especially micro-single-calcification should be significant.

Objective To evaluate the influence of different surface roughness on final surface texture of glazed ceramic. Methods Sixty porcelain discs were fabricated and randomly divided into five groups (n=12), which were sequentially grounded with water-cooled sandpapers of decreasing abrasiveness (220# , 400# , 600# , 800# and 1200# grit), and then all the specimens were glazed. Before and after glazing, the surface roughness (Ra) μm of the same specimens were evaluated quantitatively with profilometer and qualitatively using scanning electron microscopy (SEM). Results The smoothest surfaces were obtained with 1 200# sandpapers griting prior to overglazing (Ra =0. 19 μm),and 220# sandpapers griting prior to overglazing displayed the roughest surfaces (Ra =0. 55 μm) ; each group displayed significant difference before and after glazing (P < 0. 05). The images obtained through SEM were consistent with the profilometer readings. Conclusions The results suggest that decreasing the surface roughness of dental ceramic prior to glazing is of great importance in order to obtain the smoothest surface after glazing.

Objective:To evaluate the application values of intraaortic palloon pumping in the treatment of acute left heart failure.Methods:From February 2013 to January 2016,102 patients in our hospital for diagnosis and treatment of acute left heart failure were selected as the research object,all the patients were equally divided into the observation group and control group with 51 patients in each group according to the random draw envelope principle.The control group was given routine drug treatment,the observation group was given intraaortic palloon pumping treatment based on the treatment in control group,the prognosis of two groups were observed and compared between two groups.Results:The total effective rate of observation group and control group were 96.1％ and 84.3 ％,respectively,which was significantly higher in the tobservation group than that of the control group (P＜0.05).The LVEF values in the observation group and control group after treatment were 65.22 ± 4.52 ％ and 58.20 ± 5.31％ respectively,which were significantly higher than those before treatment 45.29± 5.14 ％ and 45.20± 4.89 ％,and the LVEF value of observation group was significantly higher than that of the control group (P＜0.05).After 6 months' treatment,the incidence of death,refractory ischemia,malignant arrhythmia and other recent primary end points in the observation group was 5.9 ％,the control group was 21.6 ％,which was significantly higher than that of the observation group (P＜0.05).Conclusion:Aortic balloon pump implantation could promote the improvement of left ventricular function,improve the treatment effect,reduce the occurrence of recent primary end point event in the treatment of acute left heart failure.

Objective To study the effects of Xinshuai mixture on mitochondrial function of hypertrophy cardiomyocyte cell in vitro.Methods The primary neonatal rat myocardial cells were cultured to establish hypertrophic myocardial model by AngII.The levels of superoxide dismutase(SOD), methane dicarboxylic aldehyde (MDA), monoamine oxidase(MAO) and cyclooxygenase (COX) were detected by ELISA, as the mitochondrial function and oxidative stress damage.The expression level of caspase-3 mRNA was detected by Real-time PCR.Results Compared with control group, the SOD level in model group reduced markedly, MDA level increased significantly, the activities of MAO increased, the activities of COX decreased, the expression of caspase-3 level increased significantly(P<0.01).Compared with the model group, the SOD level increased and MDA level decreased in Xinshuai mixture group and blank serum group.But the SOD level and activities of MAO, COX in Xinshuai mixture group and blank serum group had significant difference(P<0.05,P<0.01).The expression of caspase-3 mRNA level in Xinshuai mixture was significantly higher than the blank serum group(P<0.01).Conclusion Xinshuai mixture could significantly improve the fuction of mitochondrial in hypertrophy cardiomyocyte cell, reduce the energy metabolism and oxidative stress injury, and protect cardiac myocyte through lower the expression of caspase-3.

OBJECTIVE: To observe the clinical efficacy of long-time needle retaining at Baihui (GV 20) on post-stroke cognitive disorder of deficiency and blood stasis. METHODS: A total of 70 patients (3 cases dropping) with post-stroke cognitive disorder of deficiency and blood stasis were randomized into an observation group (34 cases) and a control group (33 cases). Neurological routine treatment and western medicine rehabilitation therapy were given in the control group as the basic treatment, on the basis of the treatment as the control group, long-time needle retaining at Baihui (GV 20) was applied in the observation group. The treatment was given once every other day, 3 times a week for a total of 4 weeks. The National Institute of Health stroke scale (NIHSS), mini-mental state examination (MMSE), Montreal cognitive assessment (MoCA) and deficiency blood stasis syndrome scale score were observed before and after treatment, and the clinical efficacy was compared between the two groups. RESULTS: After 4 weeks of treatment, the NIHSS scores of the two groups were lower than those before treatment (both <0.01), and the MMSE and MoCA scores were higher than those before treatment (<0.05, <0.01), and the scores in the observation group were better than those in the control group (all <0.01). After treatment, the scores of deficiency and blood stasis syndrome were lower than those before treatment (all <0.01). The scores of the shortness of breath, facial color, tongue pulse and dizziness in the observation group were better than those in the control group (all <0.05), there was no significant difference in the degree of spontaneous sweating between the two groups (>0.05). The total effective rate was 94.1% (32/34) in the observation group, which was higher than 75.8% (25/33) in the control group (<0.05). CONCLUSION Long-time needle retaining at Baihui (GV 20) is safe and effective in treating with post-stroke cognitive disorder of deficiency and blood stasis.
Cognition ; Cognition Disorders ; etiology ; therapy ; Humans ; Qi ; Stroke ; complications ; Treatment Outcome

OBJECTIVE: To evaluate the effect of curcumin on renal mitochondrial oxidative stress, nuclear factor-κB/NOD-like receptor protein 3 (NF-κB/NLRP3) inflammatory body signaling pathway and tissue cell injury in rats with acute respiratory distress syndrome (ARDS). METHODS: A total of 24 specific pathogen free (SPF)-grade healthy male Sprague-Dawley (SD) rats were randomly divided into control group, ARDS model group, and low-dose and high-dose curcumin groups, with 6 rats in each group. The ARDS rat model was reproduced by intratracheal administration of lipopolysaccharide (LPS) at 4 mg/kg via aerosol inhalation. The control group was given 2 mL/kg of normal saline. The low-dose and high-dose curcumin groups were administered 100 mg/kg or 200 mg/kg curcumin by gavage 24 hours after model reproduction, once a day. The control group and ARDS model group were given an equivalent amount of normal saline. After 7 days, blood samples were collected from the inferior vena cava, and the levels of neutrophil gelatinase-associated lipocalin (NGAL) in serum were determined by enzyme-linked immunosorbent assay (ELISA). The rats were sacrificed, and kidney tissues were collected. Reactive oxygen species (ROS) levels were determined by ELISA, superoxide dismutase (SOD) activity was detected using the xanthine oxidase method, and malondialdehyde (MDA) levels were determined by colorimetric method. The protein expressions of hypoxia-inducible factor-1α (HIF-1α), caspase-3, NF-κB p65, and Toll-like receptor 4 (TLR4) were detected by Western blotting. The mRNA expressions of HIF-1α, NLRP3, and interleukin-1β (IL-1β) were detected by reverse transcription-polymerase chain reaction (RT-PCR). Renal cell apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL). The morphological changes in renal tubular epithelial cells and mitochondria were observed under a transmission electron microscope. RESULTS: Compared with the control group, the ARDS model group exhibited kidney oxidative stress and inflammatory response, significantly elevated serum levels of kidney injury biomarker NGAL, activated NF-κB/NLRP3 inflammasome signaling pathway, increased kidney tissue cell apoptosis rate, and renal tubular epithelial cell damage and mitochondrial integrity destruction under transmission electron microscopy, indicating successful induction of kidney injury. Following curcumin intervention, the injury to renal tubular epithelial cells and mitochondria in the rats was significantly mitigated, along with a noticeable reduction in oxidative stress, inhibition of the NF-κB/NLRP3 inflammasome signaling pathway, and a significant decrease in kidney tissue cell apoptosis rate, demonstrating a certain dose-dependency. Compared with the ARDS model group, the high-dose curcumin group exhibited significantly reduced serum NGAL levels and kidney tissue MDA and ROS levels [NGAL (μg/L): 13.8±1.7 vs. 29.6±2.7, MDA (nmol/g): 115±18 vs. 300±47, ROS (kU/L): 75±19 vs. 260±15, all P < 0.05], significantly down-regulated protein expressions of HIF-1α, caspase-3, NF-κB p65, and TLR4 in the kidney tissue [HIF-1α protein (HIF-1α/β-actin): 0.515±0.064 vs. 0.888±0.055, caspase-3 protein (caspase-3/β-actin): 0.549±0.105 vs. 0.958±0.054, NF-κB p65 protein (NF-κB p65/β-actin): 0.428±0.166 vs. 0.900±0.059, TLR4 protein (TLR4/β-actin): 0.683±0.048 vs. 1.093±0.097, all P < 0.05], and significantly down-regulated mRNA expressions of HIF-1α, NLRP3, and IL-1β [HIF-1α mRNA (2^-ΔΔCt): 2.90±0.39 vs. 9.49±1.87, NLRP3 mRNA (2^-ΔΔCt): 2.07±0.21 vs. 6.13±1.32, IL-1β mRNA (2^-ΔΔCt): 1.43±0.24 vs. 3.95±0.51, all P < 0.05], and significantly decreased kidney tissue cell apoptosis rate [(4.36±0.92)% vs. (27.75±8.31)%, P < 0.05], and significantly increased SOD activity (kU/g: 648±34 vs. 430±47, P < 0.05). CONCLUSIONS Curcumin can alleviate kidney injury in ARDS rats, and its mechanism may be related to the increasing in SOD activity, reduction of oxidative stress, and inhibition of the activation of the NF-κB/NLRP3 inflammasome signaling pathway.
Male ; Rats ; Animals ; Rats, Sprague-Dawley ; NF-kappa B ; Actins ; Caspase 3 ; Curcumin ; Lipocalin-2 ; Toll-Like Receptor 4 ; Inflammasomes ; NLR Family, Pyrin Domain-Containing 3 Protein ; Reactive Oxygen Species ; Saline Solution ; Kidney ; Superoxide Dismutase