Objective To evaluate the effects of morphine preconditioning on the expression of microRNAs (miRNAs) during hypoxia-reoxygenation (H/R) in isolated cardiomyocytes in rats with heart failure.Methods Healthy adult male Sprague Dawley rats,w eighing 200-220 g,were used in this study.Adriamycin 2.0 mg/kg was injected once a week for 6 weeks via the tail vein to induce heart failure.The cardiomyocytes were isolated from the failing hearts of rats and seeded in 24-well plates or in 60 mm diameter dishes.The cells were then randomly divided into 3 groups (n =16 each) using a random number table:control group (group C); group H/R;morphine preconditioning group (group MP).The cells were cultured in normal culture atmosphere in group C.After being exposed to hypoxic air (5％ CO2-95％ N2) for 90 min,the cells were returned to the high-glucose DMEM supplemented with 10％ newborn bovine serum and were then cultured for 120 min in H/R and MP groups.In group M,the cells were cultured in morphine culture medium (final concentration of morphine 0.3 μmol/L) for 10 min and then were returned to the culture medium without morphine and cultured for 30 min immediately before hypoxia.At 120 min of reoxygenation,the cells of 8 wells in each group were chosen to detect the cell viability and lactate dehydrogenase (LDH) activity (by Typan blue staining).All the RNAs were extracted from the cardiomyocytes of the left 8 wells in each group and subjected to miRNA microarray to screen differentially expressed miRNAs.Results The cell viability was significantly lower,the activity of LDH was higher,the expression of miR-6216 and let7e-5p was higher,and the expression of miR-133b-5p was lower in H/R and MP groups than in group C (P ＜ 0.05).Compared with H/R group,the cell viability was significantly increased,the activity of LDH was decreased,the expression of miR-133b-5p was up-regulated,and the expression of miR-6216 and let-7e-5p was down-regulated in MP group (P ＜ 0.05).Conclusion Morphine preconditioning reduces H/R injury to isolated cardiomyocytes in rats with heart failure through regulating the expression of miRNAs such as miR133b-5p,miR-6216 and let-7e-5p.

OBJECTIVE: To explore the relationship between polymorphism of apolipoprotein E (ApoE) exon 4 gene and different syndromes in patients with coronary heart disease (CHD). METHODS: Two hundred patients with CHD were divided into four groups according to syndrome differentiation, including syndrome of phlegm (PS), syndrome of blood stasis (BSS), syndrome of phlegm-blood stasis blocking (PBBS) and syndrome of non-phlegm and non-blood stasis (NPNBS). One hundred healthy volunteers were included in control group. Blood lipids were measured by routine examination. Total DNA of peripheral blood was extracted. ApoE genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. All data were analyzed by SAS software. RESULTS: (1)The occurrence rate of epsilon4 allele of ApoE in patients with CHD was 19.5%, significantly higher than 9.5% in the control group (P<0.05), and the E 3/4 genotype was especially more frequent (P<0.01). (2) The levels of total cholesterol (TC), total triglycerides (TG) and low-density lipoprotein cholesterol (LDL-C) in patients with epsilon4 were higher than those in patients without epsilon4 (P<0.01). (3) The frequencies of epsilon4 allele and E3/4 genotype in patients with PS were significantly higher than those in patients with BSS (P<0.05). CONCLUSION: ApoE epsilon4 allele, especially E3/4 genotype, is the risk factor of CHD. There is a relatively close relationship between patients with ApoE epsilon4 allele and PS. It may be one of the main susceptible genes in CHD patients with PS.

AIM:To observe the role of calcium-sensing receptor (CaSR) in the regulation of pulmonary artery tension.METHODS:The intracellular calcium concentration ([Ca2+]i) was detected by laser-scanning confocal micros-copy, and the pulmonary artery tension was determined by the pulmonary arterial ring technique .RESULTS: Increased levels of [Ca2+]o or Gd3+(an agonist of CaSR) induced the increase in [Ca2+]i and pulmonary artery constriction in a concentration-dependent manner.Additionally, the effects of Ca2+and Gd3+were inhibited by U73122 and D609 (specific inhibitor of PLC), and 2-APB and heparin (specific antagonist of IP3 receptor).However, U73343 (U73122 inactive ana-logue) did not take effect.CONCLUSION: CaSR may be involved in the regulation of pulmonary artery tension by in-creasing [Ca2+]i through G-protein-PLC-IP3 pathway.

Objective To verify whether the periodic or continuous exposure to high glucose may have different effects on human umbilical vein endothelial cell (HUVEC)apoptosis, and to explore the effect of NF-κB pathway on apoptosis of HUVEC induced by high glucose using the RNAi adenovirus vector. Methods RNAi combinant adenovirus vector which targeted 1566 site of NF-κB p65 mRNA was constructed and the effect of p65 gene knockdown in HUVEC was detected by Western blot analysis. Then, the RNAi adenovirus was transducted to explore the role of NF-κB pathway on the regulation of apoptosis in HUVEC induced by high glucose. The apoptosis of HUVEC was tested by flow cytometry and TUNEL assay. Results High glucose could induce apoptosis of HUVEC. p65 protein expression of nuclear extracts was significantly increased in high glucose culture as compared to control group, but only slightly increased in NF-κB-specific knockdown group, which maintained at basal state. Compared with normal glucose group, the number of TUNEL-positive cells in high glucose group was significantly increased (25.81%±1.77% vs 8.20%±0.63%, P<0.05). The number of TUNEL-positive cells was decreased in 30.5 rmnol/L glucose plus Ad-1566 than that in 30.5 mmol/L glucose plus Ad-DEST (11.49%±0.92% vs 26.10%±0.98%, P<0.01). Flow cytometry and TUNEL assay showed that the apoptosis of human umbilical vein endothelial cells induced by high glucose was inhibited by the RNAi adenovirus. Conclusion High glucose induces apoptosis of HUVEC. Knockdown of NF-κB p65 may protect HUVEC from apoptosis by preventing high glucose-induced NF-κB nuclear translocation.

Objective To construct RNAi combinant adenoviral expressive vectors specific to p65 subunit of NF-?B and to observe their gene silencing effect on p65 subunit.Methods Three pairs of complementary. single-strand DNA oligos targeting three various sites of p65 mRNA were designed and synthesized.Annealling was used to generate double-strand oligos(ds-oligos),and then the ds-oligos were cloned into pENTR~TM/u6 to generate the entry clone named pENTR.Recombination reaction in vitro with the pENTR and pAd/BLOCK-iT~TM- DEST was used to creat the adenovirus plasmid which contains the RNAi cassette.Then,the adenovirus plasmids digested with PacI were transfected into HEK293A cells to product adenovirus,and latter infected the HEK293A cells to amplify the adenoviral stock.Plaque forming assay was used to titer the adenoviral stock.The p65 gene silencing effect induced by the RNAi adenovirus was detected by Western blot and immunocytochemistry assay in ECV304 cells.Results The RNAi adenovirus specific to p65 subunit of NF-?B were produced with titer of 3.0 x 10~9pfu/ml to 2.5?10~10pfu/ml.The expression of p65 protein in ECV304 cells could be down-regulated efficiently by the RNAi adenovirus 48-72 h after infection,which would last for more than 6 days after infection.Conclusion RNAi adenovirus is an important tool inhibiting the expression of target gene efficiently.

Adolescent ; Adult ; Aged ; Bronchiolitis ; complications ; diagnosis ; therapy ; Female ; Humans ; Male ; Middle Aged

Knowledge Discovery in Databases is gaining attention and raising new hopes for traditional Chinese medicine (TCM) researchers. It is a useful tool in understanding and deciphering TCM theories. Aiming for a better understanding of Chinese herbal property theory (CHPT), this paper performed an improved association rule learning to analyze semistructured text in the book entitled Shennong's Classic of Materia Medica. The text was firstly annotated and transformed to well-structured multidimensional data. Subsequently, an Apriori algorithm was employed for producing association rules after the sensitivity analysis of parameters. From the confirmed 120 resulting rules that described the intrinsic relationships between herbal property (qi, flavor and their combinations) and herbal efficacy, two novel fundamental principles underlying CHPT were acquired and further elucidated: (1) the many-to-one mapping of herbal efficacy to herbal property; (2) the nonrandom overlap between the related efficacy of qi and flavor. This work provided an innovative knowledge about CHPT, which would be helpful for its modern research.
Data Mining ; Drugs, Chinese Herbal ; Humans ; Materia Medica ; Medicine, Chinese Traditional ; Qi

OBJECTIVETo investigate the apoptosis induced by manumycin in U937 and HL-60 cell lines, and to explore the role of mitochondria apoptotic pathway in manumycin-inducing apoptosis.

METHODSLeukemic cells line U937 and HL-60 were treated by manumycin at 2 micromol/L for different time. Apoptosis of leukemia cells was detected by flow cytometry. The cytosolic proteins were extracted using a digitonin buffer. The protein expression of cytochrome C, caspase-9, caspase-8, and caspase-3 were determined by western blot. Mitochondrial membrane potential was detected by JC-1.

RESULTSIn U937 and HL-60 cells, manumycin induced mitochondrial depolarization after 6 h treatment. The average red/green fluorescence ratios at 6 h were significantly (P < 0.01) lower than those at time 0, being 0.51 +/- 0.07 and 0.41 +/- 0.06 for control group respectively. Manumycin induced cytochrome C release from the mitochondria into the cytosol after 6 h treatment, and activated caspase-9, caspase-8, and caspase-3 after a 16h treatment. The broad-spectrum caspase-inhibitor Z-VAD-fmk at 50 micromol/L was able to inhibit caspase cleavage completely, but only reduced the manumycin-induced apoptosis rates by 51.69% and 56.47% in U937 and HL-60, respectively.

CONCLUSIONManumycin induced apoptosis in U937 and HL-60 cell lines via mitochondria apoptotic pathway.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cytochromes c ; metabolism ; HL-60 Cells ; Humans ; Mitochondria ; drug effects ; metabolism ; physiology ; Polyenes ; pharmacology ; Polyunsaturated Alkamides ; pharmacology

Objective To learn the current status of family doctor service at pilot communities in Guangzhou, and discover existing problems and influencing factors by investigating the residents who have contracted such service , those have not and the family doctors . Methods This study chose typical community health centers of six communities in Guangzhou in January 2016 .In random sampling , residents who visited doctors during the survey and all the family doctors were surveyed .EpiData was used to doubly inputdata,withSPSS20.0forstatisticalanalysis.Results 66.0％ofthoseresidentswhohavenot contracted the service are willing to contract a family doctor .According to the binary logistic regression analysis after eliminating the interference factors , there are two factors affecting their willingness:gender and whether needing a family doctor for themselves and their family for health management .According to the binary logistic regression analysis after eliminating the interference factors , the influencing factors of renewing contract are overall satisfaction and necessity for signing family doctors .Conclusions The smooth development of the family doctor service is faced with many bottlenecks , while improving willingness to contract and renew contract to family doctors are the cornerstone for sustainability of the family doctor system.

OBJECTIVETo observe the regional distribution of propofol in canine spinal cord under noxious stimulation.

METHODSTwelve healthy hybrid dogs (12-18 months old, weighing 10-12 kg) were randomly divided into control group (n=6) and stimulation group (n=6). All the dogs were anesthetized with a single bolus dose of propofol (7 mg/kg) in 15 seconds followed by propofol infusion at a constant rate of 70 mg/kg/h via the great saphenous vein of the right posterior limb. In the stimulation group, the tails of the dogs were clamped for 5 min after 45 min of propofol infusion. Blood samples were taken from the internal carotid artery and internal jugular vein at 50 min after propofol infusion to detect plasma propofol concentrations by high-pressure liquid chromatography (HPLC). The dogs were then immediately sacrificed by decapitation and the frontal horn, posterior horn, intermediate zone, frontal funiculus, posterior funiculus and lateral funiculus of the spinal cord were dissected for determination of propol content by HPLC.

RESULTSThe plasma concentrations of propofol in the internal carotid artery and internal jugular vein were 5.07-/+0.23 and 5.03-/+0.10 microg/ml in the stimulation group, respectively showing no significant differences from those in the control group (5.09-/+0.03 and 5.08-/+0.03 microg/ml, P>0.05). In the control group, the propofol concentration was 5.09-/+0.08 microg/g in the frontal horm, 5.10-/+0.08 microg/g in the posterior horn, 5.05-/+0.19 microg/g in the intermediate zone, 5.06-/+0.14 microg/g in the frontal funiculus, 5.06-/+0.15 microg/g in the posterior funiculus and 5.06-/+0.41 microg/g in the lateral funiculus, showing no significant differences (P>0.05). The propofol concentrations in the frontal horn (7.65-/+0.47 microg/g) and posterior funiculus (7.06-/+0.82 microg/g) in the stimulation group were significantly higher than those in the other spinal cord tissues (P<0.05) and those in the control group (P<0.05).

CONCLUSIONAt 50 min after intravenous injection of propofol at a constant rate of 70 mg/kg/h, plasma propofol concentrations in the internal carotid artery and internal jugular vein reaches equilibrium with a balanced distribution in all the spinal cord regions. Propofol concentration can be higher in the frontal horn and posterior funiculus under noxious stimulation.

Animals ; Dogs ; Female ; Male ; Nociceptors ; drug effects ; physiology ; Pain ; physiopathology ; Physical Stimulation ; Propofol ; administration & dosage ; pharmacokinetics ; pharmacology ; Random Allocation ; Spinal Cord ; metabolism