Alectinib, an inhibitor of anaplastic lymphoma kinase (ALK), was approved by the Food and Drug Administration (FDA) for the treatment of patients with ALK-positive non-small cell lung cancer (NSCLC). Here we investigated the reversal effect of alectinib on multidrug resistance (MDR) induced by ATP-binding cassette (ABC) transporters, which is the primary cause of chemotherapy failure. We provide the first evidence that alectinib increases the sensitivity of ABCB1- and ABCG2-overexpressing cells to chemotherapeutic agents in vitro and in vivo. Mechanistically, alectinib increased the intracellular accumulation of ABCB1/ABCG2 substrates such as doxorubicin (DOX) and Rhodamine 123 (Rho 123) by inhibiting the efflux function of the transporters in ABCB1- or ABCG2-overexpressing cells but not in their parental sensitive cells. Furthermore, alectinib stimulated ATPase activity and competed with substrates of ABCB1 or ABCG2 and competed with [125I] iodoarylazidoprazosin (IAAP) photolabeling bound to ABCB1 or ABCG2 but neither altered the expression and localization of ABCB1 or ABCG2 nor the phosphorylation levels of AKT and ERK. Alectinib also enhanced the cytotoxicity of DOX and the intracellular accumulation of Rho 123 in ABCB1-overexpressing primary leukemia cells. These findings suggest that alectinib combined with traditional chemotherapy may be beneficial to patients with ABCB1- or ABCG2-mediated MDR.
Adenosine Triphosphatases ; Carcinoma, Non-Small-Cell Lung ; Doxorubicin ; Drug Resistance, Multiple* ; Drug Therapy ; Humans ; In Vitro Techniques* ; Leukemia ; Lymphoma ; Parents ; Phosphorylation ; Phosphotransferases ; Rhodamine 123 ; United States Food and Drug Administration

Along with the increase of clinical application, the safety of traditional Chinese medicine gained more and more attentions. In particular, the safety evaluation of Chinese medical injections has become a mandatory task should be completed by pharmaceutical companies under the supervision of China Food and Drug Administration(CFDA). Due to the weak foundation of previous studies, the safety issues of Chinese medical injections have not been fully understood, and lack of scientific and rational risk management programs. Clinical safety centralized monitoring(CSCM) is an important method for post-market safety evaluation of Chinese medicine. Due to the lack of appropriate norms and procedures, the quality of similar research is uneven, and the results vary. Combined with practical experience with experts' suggestions, we developed this expert consensus on the design and implementation of CSCM from three stages (design, implementation and report) with 20 technical points, which will provide technical support for future CSCM studies.

OBJECTIVETo summarize the clinical characteristics of peripheral blood, immune phenotypes, fusion genes and cytogenetics of patients with t(8;21) acute myeloid leukemia(AML) through the retrospective analysis of 586 patients with t(8;21) AML from 15 blood disease research centers in Northern area of China.

METHODSThe factors affecting prognosis of patients with t(8;21) AML were investigated by using univariate and multivariate COX regression.

RESULTSThe immune type of t(8;21) AML patients was mainly with HLA-DR, CD117, CD34, MPO, CD38, CD13and CD33(>95%), part of them with CD19and CD56; the most common accompanied mutation of t(8;21) AML patients was C-KIT mutation (37.8%); in addition to t(8;21) ectopic, the most common chromosomal abnormality was sex chromosome deletions (38.9%). The univariate analysis revealed a significant survival superiority of OS and PFS in t(8;21) AML patients of WBC≤3.5×10/L without C-KIT mutation, the newly diagnosed ones achieved HSCT(P<0.05), only survival superiority on OS in t(8;21) AML patients with extramedullary infiltration and CD19 positive; the results of multivariate analysis showed a significant survival superiority on OS and PFS in t(8;21) AML patients with WBC≤3.5×10/L(P<0.05).

CONCLUSIONThe clinical features of t(8;21) AML patients in China are similar to those in other countries, WBC≤3.5×10/L is a good prognostic factor while the C-KIT mutation is a poor one in t(8;21) AML patients.

OBJECTIVETo investigate the clinical outcomes of limited open reduction and percutaneous K-wire internal fixation for the treatment of irreducible Gartland type III humerus supracondylar fracture in children.

METHODSFrom May 2006 to October 2014, 132 patients with irreducible Gartland type III humerus supracondylar fracture were treated with reduction and percutaneous K-wire internal fixation. The reduction was performed with the guiding of surgeon's finger, and the lateral approach with periosteum torn was chosen according to the shift direction of the distal fractures. Among them, there were 82 males and 50 females with an average age of 5.8 years old(ranged from 2 to 14 years old).

RESULTSAll the patients were followed up, the duration ranged from 6 to 36 months, with an average of 13.7 months. Ninety-five patients got an excellent result, 27 good, 8 fair, and 2 poor.

CONCLUSIONSLimited open reduction and percutaneous K-wire internal fixation for the treatment of irreducible Gartland type III humerus supracondylar fracture in children has many advantages: simple manipulate, not affected by the elbow swelling, and satisfactory curative effect. It is worth popularizing in clinic.

OBJECTIVETo study effects of 50 Hz 1.8 mT sinusoidal electromagnetic fields (SEMFs) on bone mineral density (BMD) in SD rats.

METHODSThirty SD rats weighted(110±10) and aged 1 month were randomly divided into control group and electromagnetic field group, 15 in each group. Normal control group of 50 Hz 0 mT density and sinusoidal electromagnetic field group of 50 Hz 1.8 mT were performed respectively with 1.5 h/d and weighted weight once a week, and observed food-intake. Rats were anesthesia by intraperitoneal injection and dual energy X-ray absorptiometry were used to detect bone density of whole body, and detected bone density of femur and vertebral body. Osteocalcin and tartrate-resistant acid phosphatase 5b were detected by ELSA; weighted liver, kidney and uterus to calculate purtenance index, then detected pathologic results by HE.

RESULTSCompared with control group, there was no significant change in weight every week, food-intake every day; no obvious change of bone density of whole body at 2 and 4 weeks, however bone density of whole body, bone density of excised femur and vertebra were increased at 6 weeks. Expression of OC was increased, and TRACP 5b expression was decreased. No change of HE has been observed in liver, kidney and uterus and organic index.

CONCLUSIONS50 Hz 1.8 mT sinusoidal electromagnetic fields could improve bone formation to decrease relevant factors of bone absorbs, to improve peak bone density of young rats, in further provide a basis for clinical research electromagnetic fields preventing osteoporosis foundation.

OBJECTIVETo establish a new inducing system for differentiation of human embryonic stem (ES) cells to dendritic cells (DC), and further explore how microRNA-223 (miR-223) regulates DC differentiation from ES cells.

METHODSHuman ES cells were cultured on plates coated by IV type collagen and differentiated into hematopoietic stem/progenitor cells, common myeloid progenitor cells and DC step by step via adding different hematopoietic growth factors. The differentiated cells were identified by morphology, flow cytometry and hematopoietic colony forming unit (CFU) assays. Human ES cells were transfected with lentiviral vectors to overexpress miR-223 or inhibit miR-223 expression, then initiated the differentiation to DC. The differentiated cells at the different miR-223 levels were compared by the numbers of hematopoietic CFU and the expressions of specific surface markers. Dual-luciferase reporter assay was performed to test whether miR-223 directly targets TGFBR3.

RESULTSHuman ES cells were successfully induced into DC as the percentage of CD83 was approximately 82%, and the expression of miR-223 was up-regulated during the whole process. Supplementing miR-223 level using synthetic miR-223 mimics improved the proportions of CD34CD45, CD34CD45and CD83in differentiated cells, which were significantly higher than those in synthetic miR-223 inhibitor group and negative control (P<0.05). The expressions of cell makers at each differentiated phase in miR-223 inhibitor group were significantly lower than those in negative control (P<0.05). The differentiated cells in miR-223 mimics group showed approximately 759 CFUs per 10cells, which was significantly higher than that in others (P<0.05). Compared with negative control, miR-223 substantially inhibited the luciferase activity of Tgfbr3 3'UTR construct (by 37%) (P<0.05). In addition, the luciferase activity of the mutant construct was significantly higher than that of the WT construct in the presence of miR-223 mimics (P<0.05). With DC mature, the protein level of TGFBR3 gradually decreased using miR-223 mimics, and increased in miR-223 inhibitor group due to the suppression of the endogenous miR-223.

CONCLUSIONMiR-223 promotes the differentiation of human ES cells to DC, probably through direet target to TGFBR3.

OBJECTIVETo understand the relationships between CDH13 (T-cadherin) genetic polymorphisms, adiponectin levels and ischemic stroke, and possible interactions between CDH13 polymorphisms and other risk factors.

METHODSWe recruited 342 Chinese ischemic stroke sib pairs. We genotyped rs4783244 and rs7193788 on CDH13 using time-of-flight mass spectrometry genotyping technology and measured total and high-molecular weight (HMW) adiponectin levels. We investigated associations between SNPs and ischemic stroke, and interactions between SNPs and other risk factors using multi-level mixed-effects regression model.

RESULTSIn individuals without ischemic stroke, CDH13 rs4783244 was associated with total adiponectin levels (per T: Coef = -0.257, P = 0.001). CDH13 rs7193788 was associated with total adiponectin levels (per A: Coef = -0.221, P = 0.001) and HMW adiponectin levels (per A: Coef = -0.163, P = 0.003). rs7193788 was significantly associated with ischemic stroke (GA/AA vs. GG: OR = 1.55, 95% CI: 1.07 to 2.24, P = 0.020) after Bonferroni correction (α = 0.025). There was an interaction between rs7193788 and diabetes (P = 0.036). Compared to diabetes-free individuals with rs7193788 GG genotype, diabetes patients with rs7193788 GA/AA genotypes had higher risks for ischemic stroke (OR = 2.64, 95% CI: 1.58-4.40, P < 0.001).

CONCLUSIONCDH13 genetic polymorphisms are associated with adiponectin levels and ischemic stroke. An interaction is found between CDH13 SNP and diabetes for ischemic stroke.

Adiponectin ; blood ; Aged ; Brain Ischemia ; blood ; genetics ; Cadherins ; genetics ; China ; Female ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Risk Factors ; Stroke ; blood ; genetics

BACKGROUNDAlthough obstructive sleep apnea (OSA) has been recognized as a major risk factor for cardiovascular complications and its clinical features are well characterized, it is difficult to replicate the OSA hypoxic model in humans. We aimed to establish an experimental rabbit model for chronic OSA and to explore its application to measure blood pressure (BP), myocardial systolic function, and oxidative stress.

METHODSThe rabbit model for OSA was established by repeatedly closing the airway and then reopening it. A tube specially designed with a bag that could be alternately inflated and deflated according to a predetermined time schedule, resulting in recurrent airway occlusions and chronic intermittent hypoxia (CIH) imitating OSA patterns in humans, was used. Twenty-four rabbits were randomly divided into obstruction, sham, and control groups, and their upper airways were alternately closed for 15 s and then reopened for 105 s in a 120-s-long cycle, for 8 h each day over 12 consecutive weeks. Before and after the experiment, the BP of each rabbit was monitored. Levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the serum, superoxide dismutase (SOD) activity, malondialdehyde (MDA) and reactive oxygen species (ROS) contents, as well as Na+-K+-ATPase/Ca2+-ATPase activities in cardiac muscle were examined. In addition, cardiac functional parameters were measured using echocardiography.

RESULTSAfter 3 months, all rabbits in the obstruction group manifested sleepiness performance similar to that observed in OSA patients. Traces of airflow and SpO2showed that this model mimicked the respiratory events involved in OSA, including increased respiratory effort and decreased oxygen saturation. Gradually, the BP rose each month. CIH led to obvious oxidative stress and injured myocardial systolic performance. The serum levels of IL-6 and TNF-α increased significantly (64.75 ± 9.05 pg/ml vs. 147.00 ± 19.24 pg/ml and 59.38 ± 8.21 pg/ml vs. 264.75 ± 25.54 pg/ml, respectively, both P < 0.001). Compared with the sham and the control groups, myocardial activities of Na+-K+-ATPase/Ca2+-ATPase and SOD in the obstruction group decreased markedly, while ROS and MDA content increased.

CONCLUSIONSThese results show that the rabbit model for OSA simulates the pathophysiological characteristics of OSA in humans, which implies that this animal model is feasible and useful to study the mechanisms involved in the cardiovascular consequences of OSA.

Airway Obstruction ; blood ; pathology ; Animals ; Blood Pressure ; physiology ; Disease Models, Animal ; Female ; Hypoxia ; blood ; pathology ; Interleukin-6 ; blood ; Male ; Malondialdehyde ; blood ; Oxidative Stress ; Rabbits ; Reactive Oxygen Species ; blood ; Sleep Apnea, Obstructive ; blood ; pathology ; Tumor Necrosis Factor-alpha ; blood

Erigeron breviscapus injection(DZXI) and Breviscapus injection(DZSI) are two popular injections in treatment of acute ischemic stroke in China. Both of them are manufactured from a same herbal medicine, E. breviscapus, but DZXI is an herbal extract(mixture) preparation and DZSI is a pure compound injection. This article was aimed to systemically evaluate and compare their efficacy and safety in treatment of acute ischemic stroke. The randomized controlled trials(RCTs) were collected for comparing DZXI and DZSI with Salvia miltiorrhiza injection(FDI) as the medium, and they were compared with indirect Meta-analysis(ITC). Thirty-nine RCTs with 4 180 patients were included. Meta-analysis results showed that both DZXI and DZSI had better efficacy than FDI in acute ischemic stroke. In the indirect comparison, DZSI had a higher total efficacy than DZXI, with significantly statistical differences[OR=0.634, 95%CI = (0.432,0.928), P<0.000 01], but there was no significant difference in improvement of neurological deficit [MD=-1.19, 95%CI=(－3.57,1.19), P=0.953]. On the safety aspect, adverse reaction rate of DZXI was 1.14%, mainly including head swelling, fever and chills while DZSI had no significant adverse reactions. The limited evidences in this study showed that Breviscapine injection had higher total efficiency and safety than E. breviscapine injection, but due to the low quality of the included RCTs, these two medicines should be comprehensively compared in further high-quality clinical trials.

OBJECTIVETo study the effect of calcium-sensing receptor (CaSR) agonists and antagonists on the expression of CaSR in neonatal mice with persistent pulmonary hypertension (PPHN), and to clarify the role of CaSR in neonatal mice with PPHN.

METHODSForty-nine neonatal mice were randomly divided into four groups: control (n=10), hypoxia (PPHN; n=11), agonist (n=13), and antagonist (n=15). The mice in the PPHN, agonist, and antagonist groups were exposed to an oxygen concentration of 12%, and those in the control group were exposed to the air. The mice in the agonist and antagonist groups were intraperitoneally injected with gadolinium chloride (16 mg/kg) and NPS2390 (1 mg/kg) respectively once daily. Those in the PPHN and the control groups were given normal saline daily. All the mice were treated for 14 consecutive days. Hematoxylin and eosin staining and immunohistochemistry were used to observe the changes in pulmonary vessels. Laser confocal microscopy was used to observe the site of CaSR expression and measure its content in lung tissues. qRT-PCR and Western blot were used to measure the mRNA and protein expression of CaSR in lung tissues.

RESULTSCompared with the control group, the PPHN group had significant increases in the pulmonary small artery wall thickness and the ratio of right to left ventricular wall thickness (P<0.05), which suggested that the model was successfully prepared. Compared with the control group, the PPHN group had a significant increase in the mRNA and protein expression of CaSR (P<0.05), and the agonist group had a significantly greater increase (P<0.05); the antagonist group had a significant reduction in the mRNA and protein expression of CaSR (P<0.05).

CONCLUSIONSCaSR may play an important role in the development of PPHN induced by hypoxia in neonatal mice.

Animals ; Hypoxia ; complications ; Lung ; pathology ; Mice ; Myocardium ; pathology ; Persistent Fetal Circulation Syndrome ; etiology ; pathology ; Pulmonary Artery ; pathology ; RNA, Messenger ; analysis ; Receptors, Calcium-Sensing ; analysis ; genetics ; physiology