The effects of DHAA-urea,a novel dehydroabietylamine(DHAA) derivatives,on cell viability and glucose metabolism,in hypoxia and normoxia human hepatoma HepG2 cells were investigated.Hypoxia cells were achieved using DMEM containing high concentration of glucose without serum and pre-incubating of CoCl_2 (final concentration 150 μmol/L) for 24 h.The antiproliferation effect of DHAA-urea was measured by colorimetric MTT assay.The cellular ATP concentration,the lactate dehydrogenase(LDH) and glucose-6-phosphate dehydro genase (G6PD) activity were detected by their kits.It was shown that DHAA-urea markedly inhibited cell viability,cellular ATP level,LDH and G6PD activity in either aerobic or anaerobic circumstance in a dose-and time dependent manner.This suggested that DHAA-urea possibly inhibited HepG2 cells growth via the inhibition of glucolysis and glucolysis-dependent ATP depletion.DHAA-urea could be a promising candidate in the development of a novel class of agents used for human hepatocellular carcinoma.

Anelectrochemicallyreducedgrapheneoxide/goldnanoparticle-chitosan(ERGO/AuNP-CS) composite film modified glassy carbon electrode ( GCE) was constructed by directly electrochemical reduction of GO, and then assembly of AuNP-CS polycation on the surface. The surface morphologies of different modified electrodes including bare GCE, GCE/GO, GCE/ERGO and GCE/ERGO/AuNP-CS were characterized by scanning electron microscopy ( SEM ) . The differential pulse voltammetric behaviors of the electrodes were investigated, and the results indicated that the composite of ERGO/AuNP-CS exhibited excellent electrocatalytic oxidation activity to uric acid ( UA) molecule. In 0. 10 mol/L of phosphate buffer solution (pH=6. 5) with a scanning rate of 100 mV/s, the proposed composite film modified electrode showed a linear electrochemical response to UA in the range of 0 . 05-110 μmol/L with a detection limit of 12. 4 nmol/L ( S/N = 3 ). The electrode displayed good selectivity, reproducibility and stability in the determination of UA in human serum and urine samples with a recovery of 93 . 8%-104 . 1%. The detection results were agreed with those of conventional spectrophotometry and uricase Kit methods.

Objective To investigate the effect of heme oxygenase-1 ( HO-1 ) on the acute radiation-induced skin injury by gene transfer.Methods Thirty-three male SD rats were randomly divided into three groups as PBS-injected group,Ad-EGFP-injeeted group and Ad-HO-1-injected group ( n =11 ).In each group,three rats were used for determining the expression of target gene and the other rats were irradiated on the buttock skin with 40 Gy electron beam generated by a linear accelerator.Immediately after irradiation,rats were administered with a subcutaneous injection of PBS,Ad-EGFP or Ad-HO-1,respectively.Subsequently,the skin reactions were measured twice a week using the semi-quantitative skin injury scale.Results The strong positive expression of HO-1 was observed in subcutaneous dermal tissue after injection of Ad-HO-1.Compared to the PBS-injected group or the Ad-EGFP-injected group,a significant mitigation of skin injury was observed in Ad-HO-1-injected mice 14 d after irradiation (q =0.000-0.030,P ＜ 0.05 ).Conclusions HO-1 could significantly mitigate radiation-induced acute skin injury and Ad-HO-1 could be used to treat radiation-induced skin injury.

Objective To establish the reference interval for serum prostate-specific antigen (PSA) in apparent healthy men of different ages in Nanjing.Methods A total of 25 820 healthy men undergoing routine physical examinations in the First Affiliated Hospital of Nanjing Medical University from October 2013 to September 2015 were selected for the study.All of them were screened by prostate B ultrasound,excluding abnormal urinary tract diseases.The concentration of serum PSA and free prostate-specific antigen (fPSA) were measured by automatic luminescence immunoassay analyzer,and the fPSA/PSA values were calculated.The participants were divided into four groups (20～ 39,40～ 59,60～ 79 and older than 80 years old groups),then the median,5th,25th,75th and 95th percentiles of both PSA and fPSA/PSA were counted,respectively.Results The median of PSA (95th percentile ranges) of these groups by age from low to high were 0.78 (1.93),0.90 (2.93),1.34(6.60) and 2.01(11.91),respectively.The 25th to 75th percentiles were 0.55～1.11,0.61～1.36,0.77～2.51 and 0.94 ～ 4.19,respectively.The median of fPSA/PSA (95 th percentile ranges) were 0.37 (0.60),0.31 (0.56),0.28 (0.53) and 0.29(0.52),respectively.The 25th to 75th percentiles were 0.28～0.46,0.23～0.40,0.22～0.36 and 0.23～ 0.37,respectively.Among all the groups,median differences of both PSA and fPSA/PSA were statistically significant (P＜0.05),and PSA levels rise with age.PSA levels in different regions were different.Conclusion The PSA level of men under 40 years in Nanjing should be 0～2.5 ng/ml,40～60 years should be 0～4 ng/ml,while men who are above 60 years,could use 0～5 ng/ml as reference interval.

ObjectiveTocomparethedifferencesoftwostocksofguineapigs,thealbinoguineapigsandpigment guinea pigs , in establishing dyslipidemic model , to evaluate their lipid-lowering action , and to compare their properties for development of hyperlipidemia .Methods Two stocks of the 5-week-old guinea pigs were randomly divided into two groups, normal group (NC) and model group (Model).For the NC group, 12 guinea pigs were fed with normal chew .For the model group , after fed with high-fat diet for four weeks , 24 male guinea pigs were randomly grouped and treated with vehicle (VC group) and pitavastatin (Pit group) calcium, respectively, by gavage as well as received high-fat diet.Before and after modeling and pitavastatin treatment , blood samples were collected and subjected to analysis of plasma TC , TG, HDL-C and LDL-C, respectively .Results In the normal group , the blood lipid levels of albino guinea pigs were more stable than that of the pigmented pigs with the increase of age .After fed with high-fat diet , the plasma lipid levels of TC , TG and LDL-C were significantly increased in the two strains of guinea pigs , while HDL-C showed a decrease to varying degrees .Interestingly , the lipid level in the albino guinea pigs was significantly higher than that of pigment guinea pigs . And also, after drug administration for four weeks , pitavastatin treatment significantly decreased the elevated lipid level of TC, TG and LDL-C in the albino guinea pigs compared with that in the pigment guinea pigs .Conclusions The albino guinea pigs and pigment guinea pigs demonstrate certain differences in establishing dyslipidemic model and evaluating lipid -lowering pharmacodynamics .However , compared with the pigment guinea pigs , the albino guinea pigs have obvious superiority because of easy establishment of hyperlipidemia model and are more sensitive to lipid -lowering drugs .

Objective To observe the influence of lysyl oxidase(LOX)downregulation via RNAi on hypoxic metastasis of human lung cancer cell 95D and stduy its molecular mechanism.Methods LOX siRNA was used to transfect 95D cell line in normoxia (19％ O2 ).After 24-hour incubation,the cells were cultured in hypoxic incubator (0.5％ O2 ) for 24h.Real-time PCR assay was applied to detect LOX mRNA and Snail mRNA expression.Levels of Src,phosphorylation of Src (P-Src Y418 ) and Snail protein were determined by Western blot assay.Transwell chamber was used to evaluate the cellular invasion potential.Results Compared with 95D cells under normoxic conditions,hypoixa treatment increased LOX mRNA expression by 14 times and invasion ability by 2.12 times respectively.Compared with siRNA control group,LOX siRNA transfection decreased LOX mRNA expression,the invasion ability of hypoxic cells,and the protein expression of P-Src Y418 and Snail by 70％ - 75％,about 30％,and about 40％ respectively (P ＜ 0.05).However,it didn't affect the expression level of Src protein or Snail mRNA ( P ＞ 0.05).Conclusions Impaired metastatic potential of hypoxic human lung cancer cell induced by LOX downregulation is associated with reduced expression level of Src activation and Snail protein.The present data provids experimental evidence for LOX as a potential target for prevention and treatment of lung cancer metastasis under hypoxia.

The aim of this study is to investigate the anti-inflammatory effect of the adenosine derivative N6-(3-hydroxylaniline) adenosine (WS070117M1) on cigarette smoke plus LPS (lipopolysaccharide)-induced chronic obstructive pulmonary disease (COPD) in mice and its mechanism. COPD model was established by exposing male BALB/c mice to cigarette smoke and challenged with LPS inhalation. Supernatants of bronchoalveolar lavage fluid (BALF) were harvested and IL-1β, IL-6, IL-8 and TGF-β1 levels were measured by ELISA (enzyme-linked immunesorbent assay). The number of total white blood cells and neutrophils in bronchoalveolar lavage fluid was counted separately. Lung tissue was stained with Mayer 's hematoxylin and eosin for histopathologic examination. pAMPKa protein expression and distribution of lung tissue were analyzed by immunohistochemistry method. In vitro, levels of AMPKα phosphorylation in phorbol-12- myristate-13-acetate (PMA) differentiated THP-1 cells was detected by immunohistochemistry, IL-8 level in supernatants of cigarette smoke condensate stimulating PMA differentiated THP-1 cells was measured by ELISA. The results showed that WS070117M1 treatment significantly activated AMPKa in the lung tissue. It also resulted in down regulation of IL-1β, IL-6, IL-8 and TGF-β1 levels in bronchoalveolar lavage fluid and IL-8 level in cigarette smoke condensate stimulating PMA differentiated THP-1 cells. In addition, WS070117M1 could inhibit the recruitment of total white blood cells and neutrophils. These results suggest that WS070117M1 may alleviate the airway inflammation by activating AMPK in the lung tissue.
AMP-Activated Protein Kinases ; metabolism ; Adenosine ; analogs & derivatives ; Animals ; Bronchoalveolar Lavage Fluid ; Cell Line, Tumor ; Disease Models, Animal ; Humans ; Inflammation ; drug therapy ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Leukocyte Count ; Lipopolysaccharides ; Male ; Mice ; Mice, Inbred BALB C ; Neutrophils ; cytology ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; Smoke ; adverse effects ; Tobacco ; Transforming Growth Factor beta1 ; metabolism

The in situ gel systems can form gel in situ after administration to achieve sustained release, thus provides a promising strategy for drug delivery systems. The aim of this study was to design and prepare in situ gel systems for the oral delivery of ibuprofen (IBU-ISG) and study its pharmacokinetics in Beagle dogs. The characteristics of the basic material of gellan gum (Kelcogel, Kel) and sodium alginate (Manugel, M) were studied through investigating the complex viscosity of the Kel or M solution with or without different concentrations of calcium ion or sodium citrate to ascertain the amount range of the excipients. The measurement of complex viscosity of the solution (0. 5% Kel and 1% M) with different concentrations of sodium citrate and calcium ion was carried out to select the suitable proportion of calcium ion and sodium citrate. The formulation of binary IBU-ISG was optimized by monitoring the complex viscosity before gelling in vitro release property. The optimized formulation contains 1.0% sodium alginate, 0.5% gellan gum, 0. 21% sodium citrate and 0.056% calcium chloride. A single oral dose of IBU-ISG and reference formulation (IBU suspension) were given to each of the 6 healthy Beagle dogs, ibuprofen in plasma at different sampling times was determined by RP-HPLC. The pharmacokinetics parameters in 6 Beagle dogs were calculated. The Tmax of IBU-ISG and reference formulation were (1.8 +/- 0.6) and (0.4 +/- 0. 1) h. The Cmax values were (29.2 +/- 7.6) and (37.8 +/- 2.2) microg x mL(-1). The T(1/2) were (2.3 +/- 0.5) and (2.0 +/- 0.9) h, and the AUC(0-t) were (131.0 +/- 38.6) and (117.3 +/- 23.1) microg x mL(-1) x h, respectively. The binary IBU-ISG was successfully prepared.
Administration, Oral ; Alginates ; chemistry ; Analgesics, Non-Narcotic ; administration & dosage ; blood ; pharmacokinetics ; Animals ; Area Under Curve ; Calcium Chloride ; chemistry ; Citrates ; chemistry ; Delayed-Action Preparations ; Dogs ; Drug Compounding ; methods ; Drug Delivery Systems ; Excipients ; Female ; Glucuronic Acid ; chemistry ; Hexuronic Acids ; chemistry ; Ibuprofen ; administration & dosage ; blood ; pharmacokinetics ; Male ; Polysaccharides, Bacterial ; chemistry ; Viscosity

BACKGROUNDAdvances in catheter ablation procedures for the treatment of supraventricular arrhythmias have created the need to understand better the morphological and electrophysiological characteristics of the inferior nodal extension (INE) and transitional cellular band (TCB) in the atrioventricular (AV) junctional area.

METHODSFirstly, we observed the histological features of 10 rabbit AV junctional areas by serial sections under light microscopy. Then we recorded the action potentials (APs) of transitional cells (TCs) in the INE, TCBs, AV node, and ordinary right atrial myocytes from the AV junctional area of 30 rabbits using standard intracellular microeletrode techniques.

RESULTSUnder light microscopy, the INE appeared to be mostly composed of transitional cells linking upward to the AV node. Four smaller TCBs originated in the orifice of the coronary sinus, the region between the septal leaflet of the tricuspid valve and the coronary sinus, the inferior wall of the left atrium, and the superior interatrial septum, respectively, all linking to the INE or the AV node. Compared with ordinary atrial myocytes, the AP of the TCs in both the INE and the TCBs had a spontaneous phase 4 depolarization (not present in ordinary atrial myocytes), with a less negative maximum diastolic potential, a smaller amplitude, a slower maximum velocity of AP upstroke, and a longer action potential duration at 50% repolarization (APD50) and at 30% repolarization (APD30). The AP characteristics of these TCs were similar to those of the AV node, except that the velocities of the phase 4 spontaneous depolarization were slower and their action potential durations at 90% repolarization (APD90) were shorter. Moreover, APD50 and APD30 of the TCs of the TCBs were shorter than in the case of TCs of the AV node.

CONCLUSIONSThe TCs of the INE and TCBs are similar to slow response automatic cells. They provide a substrate for slow pathway conduction. In addition, repolarization heterogeneity exists in the AV junctional area.

The essence of endogenous turbidity in Chinese medicine (CM) is different from cream, fat, phlegm, retention, damp, toxicity, and stasis. Along with the development of modern scientific technologies and biology, researches on the essence of endogenous turbidity should keep pace with the time. Its material bases should be defined and new connotation endowed at the microscopic level. The essence of turbidity lies in abnormal functions of zang-fu organs. Sugar, fat, protein, and other nutrient substances cannot be properly decomposed, but into semi-finished products or intermediate metabolites. They are inactive and cannot participate in normal material syntheses and decomposition. They cannot be transformed to energy metabolism, but also cannot be synthesized as executive functioning of active proteins. If they cannot be degraded by autophagy-lysosome or ubiquitin-prosome into glucose, fatty acids, amino acids, and other basic nutrients to be used again, they will accumulate inside the human body and become endogenous turbidity. Therefore, endogenous turbidity is different from final metabolites such as urea, carbon dioxide, etc., which can transform vital qi. How to improve the function of zang-fu organs, enhance its degradation by autophagy-lysosome or ubiquitin-prosome is of great significance in normal operating of zang-fu organs and preventing the emergence and progress of related diseases.
Autophagy ; Humans ; Medicine, Chinese Traditional ; Proteasome Endopeptidase Complex