Aim The purpose of this study was to compare the differential expression of 15-lipoxygenase isoenzymes in the pulmonary arteries between normoxia and hypoxia and to explore their roles in the formation of hypoxic pulmomary vasoconstriction. Method Eighteen SD rats were randomly divided into two groups(n=9):the normoxic control group breathing fresh gas and the hypoxic group breeding in animal hypoxic incubator.Immunohistochemical method,in situ hybridization and Western blot were employed to determine certain 15-lipoxygenase isoenzymes which involved in the process of hypoxic pulmonary vasoconstriction.Results ①In normoxic control group,the expression of 15-LO-1 protein was detected in the pulmonary arteries;but the expression of 15-LO-2 protein wasn’t detected.②The expression of 15-LO-1 protein in hypoxic group was much stronger than that in normoxic group (P

OBJECTIVE:To optimize the purification technology of total alkaloid from the flos of Aconitum kusnezoffii. METH-ODS:The content of total alkaloid from the flos of A. kusnezoffii was determined by acid-base titration. The purification technology of total alkaloid from the flos of A. kusnezoffii was optimized by ion resin with resin type,mass concentration of loading liquid and exchange speed as factors,maximum adsorption quantity,desorption rate and mass fraction of total alkaloid as index,and verifica-tion test was conducted. RESULTS:The optimal purification technology was as follows as type 732 cation exchange resin,mass concentration of loading liquid 0.32 g/L,exchange speed of 7 column volume(BV)/h. In validation test,the content of total alka-loid was 86.88%(RSD=0.52%,n=3),and desorption rate was 92.81%(RSD=0.40%,n=3)averagely. The extraction trans-port rate of total alkaloid from 3 batches of the flos of A. kusnezoffii was 81.76% and purification transport rate was 89.47% in av-erage. CONCLUSIONS:The established method is stable and feasible,and shows high transport rate.

The objective of this study was to investigate the correlation between the gene polymorphisms of drug metabolizing enzymes and the outcome of the first induction chemotherapy in patients with de novo acute myeloid leukemia (AML). 113 de novo AML patients were enrolled in this study. The genotypes of 11 single nucleotide polymorphisms (SNP) in drug metabolizing enzymes were detected by the SNPstream(®) Genotyping System. The correlation between the distribution of genotypes and the complete remission rate of first induction chemotherapy was analyzed by logical regression. The results showed that patients with variant genotype of CYP2D6 (rs16947) had a lower complete remission (CR) rate, as compared to those with wild type (p = 0.033, OR = 0.32, 95%CI 0.112 - 0.915); meanwhile the patients with variant genotype of GSTO2 (rs156697) had a higher CR rate as compared to those with wild type (p = 0.011, OR = 3.023, 95%CI 1.289 - 7.089). Combined analysis of the above polymorphisms, showed that patients with variant genotype of CYP2D6 and wild genotype of GSTO2 (V + W) had lower CR rates in comparison to patients with wild genotypes of both polymorphisms (p = 0.017, OR = 0.183, 95%CI 0.045 - 0.735). It is concluded that CYP2D6 (rs16947) and GSTO2 (rs156697) polymorphisms are independent factors influencing CR rates of the first induction chemotherapy in de novo AML patients.
Adolescent ; Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Child ; Cytochrome P-450 CYP2D6 ; genetics ; Female ; Genotype ; Glutathione Transferase ; genetics ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; enzymology ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Remission Induction ; Treatment Outcome ; Young Adult

OBJECTIVEThe ventral part of the medial prefrontal cortex (mPFC) plays an important role in initiation and control of voluntary movement, mood and cognition. However, after the degeneration of the nigrostriatal pathway, the neuronal activity of the ventral mPFC and the role of serotonin(1A) (5-hydroxytryptamine, 5-HT(1A)) receptors in the firing of the neurons are still unknown. The present study is to investigate the change of neuronal activity in the ventral mPFC and the effect of systemic administration of the selective 5-HT(1A) receptor antagonist WAY-100635 on the activity of the neurons in normal and 6-hydroxydopamine (6-OHDA)-lesioned rats.

METHODSSingle unit responses were recorded extracellularly with glass microelectrodes from ventral mPFC neurons in normal rats and 6-OHDA unilaterally lesioned rats in vivo.

RESULTS6-OHDA lesion of the substantia nigra pars compacta (SNc) significantly increased the firing rate with no change in the firing pattern of neurons of the ventral mPFC in rats. Systemic administration of WAY-100635 (0.1 mg/kg, i.v.) did not change the mean firing rate and firing pattern of ventral mPFC neurons in normal rats. In contrast, WAY-100635 significantly decreased the mean firing rate of the neurons in rats with 6-OHDA lesion of the SNc.

CONCLUSIONThese data suggest that the degeneration of the nigrostriatal pathway results in an increase of neuronal activity of ventral mPFC and dysfunction of 5-HT(1A) receptor.

Action Potentials ; Animals ; Disease Models, Animal ; Male ; Neostriatum ; physiology ; Neural Pathways ; drug effects ; physiology ; physiopathology ; Neurons ; drug effects ; physiology ; Parkinson Disease ; physiopathology ; Piperazines ; pharmacology ; Prefrontal Cortex ; cytology ; drug effects ; physiology ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptor, Serotonin, 5-HT1A ; metabolism ; Serotonin 5-HT1 Receptor Antagonists ; Serotonin Antagonists ; pharmacology ; Substantia Nigra ; physiology

Objective To investigate the protective effects of intravitreal injection of pigment epithelial-derived factor (PEDF) gene-modified human umbilical cord mesenchymal stem cells (PEDF-MSCs) on the pathological changes in retinal tissue of diabetic rats.Methods hUCMSCs were isolated from human umbilical cord tissue using tissue culture methods,and transfected with lentiviral vectors at a infection multiplicity of 50.Then diabetic model in rats was successfully induced by intraperitoneal injection of streptozotocin.And the rats were divided into normal control (N),PBS treatment (D1),hUCMSCs treatment (D2) and PEDF-MSC treatment (D3) group according to different treatment methods.Three months after modeling,treatment began in D1,D2 and D3 group,but N group left untreated.Two weeks after treatment,the expression of PEDF-MSCs in the eye of rats was detected by fluorescence microscopy,and HE staining was performed to observe the changes in retinal structure and the full-thickness of the retina in each group.Results The expression of CD105,CD73,CD90 was observed,while the expression of CD34,CD45,CD11b,CD19 and HLA-DR did not present.After 2 weeks of treatment,it was in the vitreous cavity not the retina that clusters of red fluorescence appeared in D2 group with fluorescence microscope.There were clusters of green fluorescence in the vitreous cavity not in the retina of D3 group.HE staining showed that the retina had intact structure and clear layers as well as neatly arranged and stained evenly cells in N group.In D1 group,the nerve fibers layer (NFL) showed obvious edema,the blood vessels were dilated,the inner plexiform layer (IPL) were loose and the inner nuclear layer (INL) cells were disordered.In D2 group,the edema of NFL relieved.In D3 group,NFL edema was significantly alleviated,and the cells of INL and outer nuclear layer (ONL) arranged in regular.Full-thickness of retina was (103.82 ±4.15) μm in N group,(138.86 ±4.71) μm in D1 group,(131.17 ±3.89) μm in D2 group,and (112.24 ±4.22) μm in D3 group,respectively,and the differences were statistically significant (all P ＜ 0.05).Conclusion PEDF-MSCs can survive and continue to express in the vitreous cavity of diabetic rats for a long time.Meanwhile,intravitreal injection of PEDF-MSCs can ameliorate retinal edema and the retinal injury in diabetic rats.

AIM:To explore the expression level of long non-coding RNA(lncRNA)myocardial infarction-as-sociated transcript(MIAT)in the tissues and cells of non-small-cell lung carcinoma(NSCLC), and to investigate the effect of MIAT on the function of NSCLC cell line.METHODS:Bioinformatic data in microarray dataset GSE19804 from Gene Expression Omnibus(GEO)were collected for analyzing the difference expression of MIAT between NSCLC tissues and normal lung tissues.Clinical and prognostic data in microarray dataset GSE 30219 from GEO were also collected for an-alyzing the correlation between the expression level of MIAT and the survival time of NSCLC patients.qPCR was applied to detect the expression of MIAT in 25 paired tumor tissues and corresponding adjacent normal tissues,normal lung epithelial HBE cell line and NSCLC A549,NCI-H266 and NCI-H1299 cell lines.The specific small interfering RNA for MIAT(si-MIAT group)or negative control sequence(si-NC group)was transfected into A549 cells,and flow cytometry,colony for-mation experiment and CCK-8 assay were employed to detect the proliferation of the cells in the 2 groups.The expression levels of cyclin D1 and cyclin-dependent kinase inhibitor 1A(CDKN1A)in the 2 groups were determined by qPCR and Western blot.RESULTS:In the GEO dataset GSE19804,the expression of MIAT in NSCLC tissues was significantly ele-vated compared with normal lung tissues(P<0.05).In the GEO dataset GSE30219,the overall survival time was signifi-cantly shorter in the patients with high expression of MIAT than the patients with low expression of MIAT(P<0.05).Fur-thermore,the levels of MIAT in both NSCLC tissues and cells were higher than those in adjacent normal tissues and normal cells(P<0.05).Compared with si-NC group,lower MIAT level,cell viability and cell colony number in si-MIAT group with statistical significance were observed(P<0.05).Meanwhile, compared with si-NC group, the expression of cyclin D1 in si-MIAT group was significantly decreased(P<0.05),and inversely,the expression of CDKN1A in si-MIAT group was significantly increased(P<0.05).CONCLUSION:There is high expression of MIAT in NSCLC tissues and NSCLC cells,and knockdown of MIAT expression inhibits NSCLC cell proliferation, which provides a potential target of targeted therapy for NSCLC.

OBJECTIVETo review theories and technologies of big data mining and their application in clinical medicine.

DATA SOURCESLiteratures published in English or Chinese regarding theories and technologies of big data mining and the concrete applications of data mining technology in clinical medicine were obtained from PubMed and Chinese Hospital Knowledge Database from 1975 to 2015.

STUDY SELECTIONOriginal articles regarding big data mining theory/technology and big data mining's application in the medical field were selected.

RESULTSThis review characterized the basic theories and technologies of big data mining including fuzzy theory, rough set theory, cloud theory, Dempster-Shafer theory, artificial neural network, genetic algorithm, inductive learning theory, Bayesian network, decision tree, pattern recognition, high-performance computing, and statistical analysis. The application of big data mining in clinical medicine was analyzed in the fields of disease risk assessment, clinical decision support, prediction of disease development, guidance of rational use of drugs, medical management, and evidence-based medicine.

CONCLUSIONBig data mining has the potential to play an important role in clinical medicine.

Bayes Theorem ; Clinical Medicine ; Data Mining ; Decision Support Systems, Clinical ; Decision Trees ; Evidence-Based Medicine ; Fuzzy Logic ; Humans ; Neural Networks (Computer) ; Pattern Recognition, Automated

In the present study, extracellular recording was used to examine the neuronal activity of the basolateral nucleus (BL) of the amygdala and the effects of systemic administration of the selective 5-HT(1A) receptor antagonist WAY-100635 on the neuronal activity in the normal rats and rats with 6-hydroxydopamine (6-OHDA)-produced lesions in the substantia nigra pars compacta (SNc). The results showed that the firing rates of BL projection neurons and interneurons were (0.39±0.04) Hz and (0.83±0.16) Hz in the normal rats, and (0.32±0.04) Hz and (0.53±0.12) Hz in 6-OHDA-lesioned rats. There was no significant difference in the firing rates of BL projection neurons and interneurons between the normal and 6-OHDA-lesioned rats. In the normal rats, all BL projection neurons fired in burst; 94% of BL interneurons fired in burst and 6% fired irregularly. In 6-OHDA-lesioned rats, 85% of BL projection neurons displayed a burst firing pattern and 15% fired irregularly; 86% of BL interneurons had a burst firing pattern and 14% fired irregularly. The distribution of firing patterns of projection neurons and interneurons in the BL in 6-OHDA-lesioned rats did not differ from that in the normal rats. Systemic administration of WAY-100635 at 0.1 mg/kg body weight did not change the mean firing rates of projection neurons and interneurons in the BL in both normal and 6-OHDA-lesioned rats. However, a higher dose of WAY-100635 at 0.5 mg/kg body weight significantly decreased the mean firing rate of BL projection neurons from (0.43±0.07) to (0.15±0.02) Hz in the normal rats (P<0.01), but significantly increased the activity of BL projection neurons in 6-OHDA-lesioned rats from (0.37±0.08) to (0.69±0.18) Hz (P<0.004). The mean firing rates of BL interneurons in the normal and 6-OHDA-lesioned rats did not change after administration of a higher dose of WAY-100635 at 0.5 mg/kg body weight. These results demonstrate that the activity of BL neurons after substantia nigra dopaminergic lesion in the SNc is regulated by activation of intrinsic and extrinsic inputs, and that 5-HT(1A) receptors significantly contribute to the regulation of the activity of BL projection neurons in both normal and 6-OHDA-lesioned rats. Furthermore, WAY-100635 induced an increase in the mean firing rate of projection neurons in the BL in 6-OHDA-lesioned rats, suggesting that 5-HT(1A) receptor is likely to play a role in generating affective symptoms in Parkinson's disease.
Action Potentials ; Amygdala ; drug effects ; Animals ; Neurons ; drug effects ; Oxidopamine ; adverse effects ; Piperazines ; pharmacology ; Pyridines ; pharmacology ; Rats ; Receptor, Serotonin, 5-HT1A ; Serotonin 5-HT1 Receptor Antagonists ; pharmacology ; Substantia Nigra ; pathology

AIMTo construct short hairpin RNA (shRNA) eukaryotic expression vectors targeting TGF-beta1 for further research on the effects of TGF-beta1 on vasculogenesis and angiogenesis.

METHODSThree pairs of siRNA target sequences coding from the mRNA of TGF-beta1 gene were designed and three pairs of nucleotides were synthesized. After annealing, the double-strand DNA products were ligated into the pEN_mH1c entry vector, and in turn into the shRNA eukaryotic expression vector pDS_hpEy labled by GFP through the LR recombination reaction. After sequencing successfully, the three resulting TGF-beta1 shRNA expression vectors were transfected into the mouse fibroblast cell line (NIH/3T3), and then cell clones stably expressing TGF-beta1 shRNA were screened. Reverse Transcript-Polymerase Chain Reaction (RT-PCR) and Western blot were used to detect the mRNA and protein expression.

RESULTSRT-PCR and Western blot showed that one of the TGF-beta1 shRNA expression vectors pDS_Tc downregulated TGF-pl mRNA and protein expression markedly in NIH/3T3 cells.

CONCLUSIONShRNA eukaryotic expression vectors targeting TGF-beta1 are successfully constructed which can be used for further investigation on the mechanism through which TGF-beta1 regulates vasculogenesis and angiogenesis.

Animals ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; NIH 3T3 Cells ; Neovascularization, Physiologic ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Transforming Growth Factor beta1 ; genetics ; metabolism

OBJECTIVETo investigate the inhibitor of matrix metalloproteinase-2 (MMP-2) (2R)-2-[5-[4-[ ethyl-methylamino] phenyl [thiophene-2-sulfonylamino]-3-methylbutyric acid (TISAM) therapeutic effect on experimental autoimmune myocarditis (EAM) in Lewis rats.

METHODSTreatment protocol of oral administration of 5 mg/kg TISAM once a day for 14 days was performed on EAM Lewis rats. EAM Lewis rats were divided into 3 groups: treatment in early, middle and later stage respectively (n = 20). After experiment at the designate time point, the rats were euthanatized and hearts were harvested. Cardiac inflammatory score, fibrosis score and content, and infiltration of macrophages and T lyminflammatory score, fibrosis score and content, and infiltration of macrophages and T lymphocytes, message RNA (mRNA) expression of matrix metalloproteinase (MMP)-2 and MMP-9 and protein activity of gelatinase were determined.

RESULTSTISAM treatment in early phase was invalid (treatment started from the creation of the model), treatment in middle and later phase was effective (treatment started from 7 and 14 day after the creation of the model).

CONCLUSIONInhibitor of MMP-2 can block ventricular remodeling in middle stage in EAM Lewis rats. The mechanism maybe alleviate the inflammatory cell cardiac infiltration, decrease the mRNA expression of MMP-2 at transcript level and downregulate gelatinase activity at protein level.

Animals ; Autoimmune Diseases ; drug therapy ; Female ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase Inhibitors ; therapeutic use ; Myocarditis ; drug therapy ; Rats ; Rats, Inbred Lew ; Thiophenes ; therapeutic use