OBJECTIVETo observe the inhibitory effect of 4-chlorobenzoyl berbamine (BBD9) on imatinib-resistant cell line K562 (K562/IR) in vitro and in vivo and explore the mechanisms.

METHODSThe IC50 of BBD9 and berbamine (BBM) was determined by MTT assay. The expressions of p210(Bcr-Abl), IKKa, cytoplasmic and nuclear NF-κBp65 were determined using Western blotting in K562/IR cells following a 48-h exposure to 0.5 µg/ml BBD9 or 8 µg/ml BBM. Flow cytometry was used to analyze the cell viability, apoptosis and necrosis; Western blotting was employed to determine the expressions of PARP, caspase-3, caspase-9 and LC3II in K562/IR cells exposed to different concentrations of BBD9 for 48 h. In nude mouse models bearing K562/IR cell xenograft, the tumor weight, tumor regression, and body weight changes of the mice were measured after treatments with 15 mg/kg and 30 mg/kg BBD9 and 100 mg/kg imatinib.

RESULTSThe IC50 of BBD9 and BBM was 0.73 µg/ml and 5.43 µg/ml, respectively. In K562/IR cell cultures, the expressions of p210(Bcr-Abl), IKKa and nuclear NF-κB p65 were all decreased following BBD9 and BBM treatments, but BBD9 produced more potent effect; cytoplasmic NF-κB p65 showed no obvious changes after the treatments. The cell apoptosis and necrosis increased with the concentrations of BBD9, which also dose-dependently increased the levels of cleaved caspase-3, csapase-9, PARP, and LC3II expression. In the tumor-bearing mouse model, BBD9 showed stronger effects than imatinib in reducing the tumor weight, promoting tumor regression, and increasing the body weight.

CONCLUSIONBBD9 can effectively inhibit the growth of K562/IR cells in vitro and in vivo by activating cell apoptosis, necrosis and autophage pathways, down-regulating expressions of p210(Bcr-Abl) and IKKa and suppressing the cytoplasm-to- nucleus translocation of NF-κBp65.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Benzamides ; Benzylisoquinolines ; pharmacology ; therapeutic use ; Drug Resistance, Neoplasm ; Female ; Fusion Proteins, bcr-abl ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; I-kappa B Kinase ; metabolism ; Imatinib Mesylate ; K562 Cells ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; Mice ; Mice, Nude ; Piperazines ; pharmacology ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; Pyrimidines ; pharmacology ; Transcription Factor RelA ; metabolism ; Xenograft Model Antitumor Assays

Objective The objective of this study was to investigate the abnormal expression of Notch signaling pathway members in pancreatic cancer and its important effect on the development of pancreatic carcinoma. Methods Affymetrix gene expres-sion microarray was used to screen the differentially expressed members of Notch signal pathway in 10 cases of pancreatic carcinoma and its adjacent tissues,and verified by real-time quantitative PCR and Western blotting. The lentivirus expression vector carrying the siRNA fragment of Jagged 2(JAG2)gene was transfected into the pancreatic cancer primary cells to construct the JAG2 gene re-pression-expressing pancreatic cancer cell line. MTT,flow cytometry and Transwell assays were used to analyze cell proliferation, changes of cell cycle and invasive transfer capabilities. Results A total of 512 differentially expressed genes were detected by Affy-metrix gene expression microarray,including 419 up-regulated genes and 93 down-regulated genes. JAG2( up-regulated expres-sion 8. 20 times),NOTCH1(up-regulated expression 3. 74 times),HES1(up-regulated expression 3. 27 times),and NOTCH2(up-regulated expression 3. 16 times)were differentially expressed in Notch signaling pathway. The results of PCR and Western blotting were consistent with those of gene chip. The growth curves of JAG2 gene repressed pancreatic cancer cells and pancreatic cancer pri-mary cells were drawn by the standard OD490 value of d1-d5 by MTT assay. JAG2 gene repressor expression vector could signifi-cantly inhibit the proliferation of pancreatic cancer cells. The cell cycle analysis showed that the apoptosis and the arrested cell cycle at the S phase were significantly increased in pancreatic cancer cells with JAG2 gene repressor expression. The invasive ability was significantly reduced in JAG2 gene repressor expression pancreatic cancer cells(P<0. 05). Conclusion Some members of the Notch signaling pathway are significantly differentially expressed in pancreatic cancer tissues,and repression of this member can affect the growth,cell cycle,invasion and metastasis of pancreatic cancer cells.

Objective Transplantation of bone marrow mesenchymal stem cells (BMSCs) has a good prospect of application for cerebral infarction,but the environment and the inflammatory response to ischemia and hypoxia after cerebral infarction are not con-ducive to the survival of transplanted cells. This article investigated the effects of Shuxue Tongmai capsule(SXTM) combined with BM-SCs transplantation on the improvement of cerebral ischemic injury in rats. Methods A model of middle cerebral artery occlusion was es-tablished in Sprague-Dawley(SD) rats using thread method and these 15 SD rats were randomly divided into model group,BMSCs group and combination therapy group (BMSCs transplantation combined with SXTM treatment). At 24h after modeling,rats in combination therapy group were given tail vein injection of 1 mL BMSCs suspension (2× 109 per/L) and gavage administration of SXTM 0. 64 g/kg. Rats in BMSCs group were given tail vein injection of 1 mL BMSCs suspension (2×109 per/L) and gavage administration of equal volume of sa-line. For model group,the rats were given tail vein injection of equal volume of PBS and gavage administration of equal volume of sa-line. Neurologic function was assessed before cell transplantation and at 3,7,14,28 days after cell transplantation to check the injury of neurologic function. At 28 days after transplantion,the rats were decapitated after anesthesia to take brain tissues for immunohisto-chemical detection of vascular endothelial growth factor(VEGF) and brain-derived neurotrophic factor(BDNF) protein expression. Mor-phological changes of the brain tissue and apoptosis in cortical neurons were observed and detected by hematoxylin-eosin staining and TUNEL,respectively. Results At 7,14,28 days after transplantation,the neurological defect score in combination therapy group was significantly lower than those of model group and BMSCs group(P<0.05). In each group,the neurological defect score at 3 days after transplantation was significantly decreased compared with those before transplantation(P<0.05). In the same group,the neurologi-cal defect scores at 14,28 days after transplantation were significantly decreased compared with those at 7 days after transplantation (P<0.05). The neurological defect scores at 14,28 days after transplantation were significantly decreased compared with those at 7 days after transplantation(P<0.05). The neurological defect score at 28 day after transplantation was significantly decreased compared with that at 7 day after transplantation(P<0.05). At 28 day after transplantation,the number of apoptotic cells in combination therapy group (51.40±4.04) was significantly fewer than those of model group (74.80±5.31) and BMSCs group (67.20±4.66) and the num-ber of apoptotic cells in BMSCs group was significantly decreased compared with model group(P<0.05). The results of immunohisto-chemistry showed that the VEGF and BDNF positive cells in the cerebral ischemic region of rats were brownish or sepia in color. Com-pared with model group,the expression levels of VEGF and BDNF protein in BMSCs group and combination therapy group were signifi-cantly increased (P<0.05),and that of combination therapy group was significantly increased compared with BMSCs(P<0.05). Conclusion SXTM combined with BMSCs transplantation can promote neurological recovery from cerebral ischemia by increasing the protein expression of VEGF and BDNF and reducing neuronal apoptosis.

BACKGROUNDFulminant type 1 diabetes (F1D) is a complex disease. Microarray analysis was used to identify gene expression changes and obtain understanding of the underlying mechanisms.

METHODSMicroarray analysis was performed on peripheral blood mononuclear cells from six F1D patients and six matched healthy subjects. Real-time polymerase chain reaction was used to verify the differentially expressed genes. NK cell activity was detected by methyl thiazoleterazolium assay.

RESULTSMicroarray analysis identified 759 genes differing in expression between F1D patients and controls at a false discovery rate of 0.05. Expression of TLR9, ELF4 and IL1RAP were verified and consistent with changes in microarray results. NK cell activity was decreased in F1D. With use of a knowledge base, differentially expressed genes could be placed within different pathways with predicted functions including interleukin-1, and tumor necrosis factor-α signaling.

CONCLUSIONSThese results identify several genes indicating possible mechanisms in F1D. NK cell dysfunction resulting from changes in expression of TLR9, ELF4 and IL1RAP, and pathways of interleukin-1 and tumor necrosis factor-α signaling might be involved in F1D through inducing β-cell dysfunction.

Adult ; Diabetes Mellitus, Type 1 ; genetics ; metabolism ; Female ; Gene Expression Profiling ; Humans ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult

Objective To explore the activity of brain default mode network (DMN) in patients with mild Alzheimer's disease (AD) by resting state functional magnetic resonance imaging (fMRI) and investigate its possible neural mechanism. Methods Twenty-four patients with mild AD,admitted to our hospital from January 2009 to June 2010,and 25 normal controls (NCs) were chosen in this study.All subjects were examined by Mini Mental State Examination (MMSE), Mattis Dementia Rating Scale (DRS) and fMRI.Resting-state whole brain data were analyzed by amplitude of low frequency fluctuation (ALFF) with two-sample t test and the brain regions in mild AD patients having significantly different ALFF comparing to NCs were observed. Results As compared with that in NCs,the memory function in mild AD patients was seriously impaired (P＜0.05).As compared with NCs,mild AD patients showed significantly decreased ALFF in the posterior cingulate cortex,ventral medial prefrontal cortex and dorsal medial prefrontal cortex, which were closely relevant to the memory (P＜0.05). Conclusion AD patients show significantly decreased active intensity of some DMN nodes that relate to memory in resting state; DMN abnormalities play an important role in early memory impairment of AD patients.

Carcinoma, Non-Small-Cell Lung ; diagnosis ; metabolism ; China ; Consensus ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; diagnosis ; metabolism ; Oncogene Proteins, Fusion ; metabolism ; Polymerase Chain Reaction ; Receptor Protein-Tyrosine Kinases ; metabolism

Objective:To investigate the availability, prices and affordability of essential medicines in pediatric population across China, in the hope of improving rational use of medicines.Methods:A multicenter cross-sectional survey of medicine prices, availability and affordability was conducted in 17 provinces, municipalities and autonomous region across east, south-central part, west and north of China. Data on 42 medicines used in pediatric population, both original and generic, were collected in 55 public hospitals from May 26 to June 2, 2017. Availability was expressed as the percentage of hospitals with stock of the target medicine on the day of data collection,and median price ratio (MPR) was the ratio of price upon investigation to international reference. Based on national minimum daily wage, affordability represents the number of working days needed to earn the expense which covers a standard course using the target medicine. Statistical software SPSS 13.0 was applied for descriptive analysis of availability, MPR and affordability.Results:Mean Availability of original and generic medicine was 33% and 32%, with median MPR being 5.43 and 1.55. Among the 19 medicines with price information for both original and generic product, the median MPR was 7.73 and 2.04 respectively. Regarding the five medicines used to treat four common pediatric diseases (pneumonia,peptic ulcer, congenital hypothyroidism, refractory nephrotic syndrome), the affordability was 0.63 (0.16-6.17) d for generic medicine, and 1.03 (0.16-11.53) d for its original counterpart.Conclusions:The availability to both original and generic products of the 42 medicines used in pediatric population was low in China. The prices of generic medicines seem to be lower and affordability higher than those of original medicines. There is an urgent need to improve the availability and affordability of pediatric medicines.

BACKGROUNDGlutamic acid decarboxylase antibody (GADA) and protein tyrosine phosphatase antibody (IA-2A) are two major autoantibodies, which exert important roles in the process of type 1 diabetes mellitus (T1D). Our study aimed to investigate the changes in positivity and titers of GADA and IA-2A during the course of Chinese acute-onset T1D patients and their relationships with clinical features.

METHODSTwo hundreds and forty-seven Chinese newly diagnosed acute-onset T1D patients were consecutively recruited. GADA and IA-2A were detected at the time of diagnosis, one year later, 3-5 years later after diagnosis during the follow-up; all the clinical data were recorded and analyzed as well.

RESULTSDuring the course of acute-onset T1D, the majority of patients remained stable for GADA or IA-2A, however, a few patients changed from positivity to negativity and fewer patients converted from negativity to positivity. The prevalence of GADA was 56.3% at diagnosis, decreasing to 50.5% one year later, and 43.3% 3-5 years later while the corresponding prevalence of IA-2A were 32.8%, 31.0% and 23.3%, respectively. The median GADA titers were 0.0825 at diagnosis, declining to 0.0585 one year later and 0.0383 3-5 years later (P < 0.001), while the corresponding median titers were 0.0016, 0.0010, 0.0014 for IA-2A, respectively. Fasting C-peptide (FCP) and postprandial C-peptide 2 hours (PCP2h) levels of GADA or IA-2A negativity persistence patients were higher than those of positivity persistence and negativity conversion patients (P < 0.05) which indicated GADA or IA-2A negativity persistence T1D patients had a less loss of β cell function.

CONCLUSIONOur data suggest that repeated detection of GADA and IA-2A are necessary for differential diagnosis of autoimmune diabetes and the indirect prediction of the β cell function in Chinese patients.

Adolescent ; Adult ; Aged ; Antibodies ; therapeutic use ; Asian Continental Ancestry Group ; Child ; Child, Preschool ; Diabetes Mellitus, Type 1 ; drug therapy ; immunology ; Female ; Glutamate Decarboxylase ; immunology ; Glycated Hemoglobin A ; metabolism ; Humans ; Infant ; Male ; Middle Aged ; Protein Tyrosine Phosphatases ; immunology ; Young Adult

OBJECTIVETo investigate the effect of simulated microgravity and carbon ion irradiation (CIR) on spermatogenic cell apoptosis and sperm DNA damage to the testis of male Swiss Webster mice, and assess the risk associated with space environment.

METHODSSperm DNA damage indicated by DNA fragmentation index (DFI) and high DNA stainability (HDS) was measured by sperm chromatin structure assay (SCSA). Apoptosis of spermatogenic cells was detected by annexin V-propidium iodide assay. Bax (the expression levels of p53) and proliferating cell nuclear antigen (PCNA) were measured by immunoblotting; p53 and PCNA were located by immunohistology.

RESULTSHDS, DFI, apoptosis index, and the expression levels of p53 and Bax were detected to be significantly higher in the experimental groups (P<0.05) compared with those in the control group; however, the PCNA expression varied to a certain degree. p53- and PCNA- positive expression were detected in each group, mainly in relation to the spermatogonic cells and spermatocytes.

CONCLUSIONThe findings of the present study demonstrated that simulated microgravity and CIR can induce spermatogenic cell apoptosis and sperm DNA damage. Sperm DNA damage may be one of the underlying mechanisms behind male fertility decline under space environment. These findings may provide a scientific basis for protecting astronauts and space traveler's health and safety.

Animals ; Apoptosis ; radiation effects ; Carbon ; Cell Proliferation ; radiation effects ; DNA Damage ; Heavy Ions ; adverse effects ; Immunohistochemistry ; Male ; Mice ; Random Allocation ; Sperm Count ; Spermatogenesis ; radiation effects ; Spermatozoa ; radiation effects ; Testis ; radiation effects ; Weightlessness Simulation

Air Pollution ; Asthma/drug therapy* ; China ; Humans ; Treatment Adherence and Compliance