OBJECTIVE:To establish a simple and rapid ion-pair-RP-HPLC method for the determination of intracellular concentration of ara-CTP.METHODS:Mononuclearcells were separated from the whole blood,centrifugated and injected for determination on Symmetry C18 column with the mobile phase consisted of 0.1 mol?L-1 KDP buffer solution and 0.01 mol?L-1 tetrabutylammonium hydrogen sulfate(pH=2.7)at a flow rate of 1.0 mL?min-1 with the UV wavelength set at 278 nm.RESULTS:The linear range of ara-CTP was 0.35-9.12 ?g?mL-1(r=0.998 6)with its detection limit at 0.35 ?g?mL-1.The recoveries of ara-CTP at low,middle and high concentrations,and the intra-day and inter-day RSDs were all up to the standards.CONCLUSION:The established method is simple,rapid,stable,reproducible and applicable for the monitoring of intracellular concentration of ara-CTP.

Objective To study application of surface plasmon resonance(SIR)system in detection of clinical pathogen with a gene chip.Methods 27 clinical samples were detected by SPR-based gene chip system.These samples were composed by 8 positive blood samples,3 positive pyoid samples,9 positive leucorrhea samples and positive reproductive tract pyoid samples,1 positive biopsy sample and 6 negative biopsy samples.Specific primers and probes for target pathogens were designed by bioinformatics methods and validated by PCR and enzyme-labelled chemiluminescence,respectively.SPR-based gene chip was prepared and utilized to detect clinical samples by SPR system.Results The primers and probes showed good specificity and accuracy,which can be applied to perform PCR and application of the gene chip.Compared with the clinical analysis,gene chip analysis of 26 clinical samples showed the consistent results.Conclusions SPR detection system proved to be accurate and reliable.The chip will have a promising prospect in application.

Objective The purpose of this study was to investigate the mechanism of action of polypeptide extracts of Eupolyphaga sinensis Walker ( ESW) against oxidative aging.Methods Mice were intraperitoneally injected D-galac-tose for consecutive 20 days to establish an aging mouse model.The model mice were administered with different doses of ESW polypeptide (0, 40, 80, 160 mg/kg/d).The normal activity, movement and anti-stress ability of the mice were ob-served.The activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) in blood and different tissues and the content of glutathione ( GSH) and malondialdehyde ( MDA) of the aging mice were assessed by xanthin oxidase activity measurement and spectrophotometry, respectively.The expression of nuclear factor erythroid 2-re-lated factor 2 (Nrf2) in Caco-2 cells was detected by immunofluorescence.Results Comparing the control and polypep-tide groups, there were significant decreases of body weight gain, organ indexes, anti-stress ability and activity capacity, the activity of SOD, CAT, GSH-PX and the content of GSH, and an increase of the content of MDA in blood and different tissues in the aging mice.With the increasing dose of polypeptide extracts of ESW, the body weight gain, organ indexes of the liver, spleen and kidney were significantly increased, the static and dynamic exercise time was prolonged in the poly-peptide group, and their abilities of hypoxia tolerance and heat tolerance were close to that of normal controls.The SOD, CAT, GSH-PX activity and GSH level in blood and different tissues were significantly increased, but MDA content de-creased.The expression of Nrf2 in Caco-2 cell nuclei was significantly increased in the polypeptide group, close to that of the positive control group.Conclusions The results of our study show that polypeptide extracts of ESW improve the anti-stress and antioxidative capacity in D-galactose-induced mouse models of oxidative aging by initiating Nrf2-ARE antioxidant signaling pathway, therefore, delay the oxidative aging in mice.

ObjectiveTo observe and detect the change of invasion and migration about CNE cell line after 5-ALA Photodynamic therapy.MethodsThe relation between the inhibition rate and 5-ALA concentration & energy density of lasers was detected by MTT.Scratch wound assay and matrigel artificial basement membrane invasion assay were perforned after photodynamic therapy with different photosensitizer concentration and energy density of lasers.ResultsThe results showed that photodynamic therapy can inhibit the growth of CNE cells.The migration of CNE cell was inhibited with appropriate photosensitizer concentration and energy density of laser after photodynamic therapy.It exhibited a linear relation between cell migration and the energy density, when the photosensitizer concentration was a maximum effective concentration of 1 mmol/L (P ＜0.05). The invasion of CNE cell was also inhibited after photodynamic therapy,there is a relationship between the change of invasion and 5-ALA concentration & energy density of lasers.It which exhibited a linear relation between cell invasion and the energy density in the same photosensitizer concentration (r ＞0.8, P ＜0.0001). ConclusionThe migration and invasion of CNE cells are both inhibited after photodynamic therapy. The change of migration and invasion is related to 5-ALA concentration & energy density of laser.

OBJECTIVE: To observe whether injection of medicine into low hydraulic resistance point along meridian brings about higher medicinal effect and to explore the efficacy of the theory that meridians are made up of channels featuring low hydraulic resistance by observing the diuretic effect of injecting furosemide or saline into the low hydraulic resistance point Shuifen (CV 9), vein and Zusanli (St 36) respectively. METHODS: Acute edema was induced in pigs by rapid intravenous injection of 2 000 ml normal saline. The pigs were divided into four groups: Shuifen (CV 9) injection of half dose furosemide group (SFF group), intravenous injection of full dose furosemide group (VF group), Zusanli (St 36) injection of full dose furosemide group (ZSLF group), and Shuifen (CV 9) injection of half dose normal saline group (SFS group). The accumulated urine quantity and the urine quantity generated in every 15-minute period were measured in each group respectively, every 15 minutes after injection, and the measurement lasted for two hours at one experiment. Each group involved eight times of experiments with one pig used for one experiment, which means the whole observation involved 32 times of experiments. RESULTS: The accumulated urine quantities observed in both SFF group and VF group were higher than those in the ZSLF group and the SFS group all through the measurement, showing significant differences during the period from the 15th minute to the 45th minute (P<0.05). But no significant difference was observed between the SFF group and the VF group during the whole 2-hour measurement (P>0.05). Analysis of urine quantity generated in every 15-minute period showed that diuretic effect climaxed during the 15th minute to the 30th minute in both SFF group and VF group. By contrast, ZSLF group reached diuresis climax during the 45th minute to 60th minute and no diuresis climax was observed in the SFS group all through the measurement. CONCLUSION: Injection of medicine into low hydraulic resistance point along meridian generates faster and more powerful medicinal potency, and this is likely to be applied to clinical practice. The theory that meridians are channels featuring low hydraulic resistance is important to the elucidation of meridians.

Objective:To investigate the values of right ventricular free wall longitudinal strain (RVFWLS) by three-dimensional speckle tracking echocardiography (3D-STE) in predicting the degree of RV myocardial fibrosis (MF) in patients with end-stage heart failure (HF).Methods:A total of 102 consecutive patients with end-stage HF undergoing heart transplantation were enrolled in the Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from June 2018 to December 2019. Echocardiographic examinations were performed in these patients before heart transplantation. The conventional RV function parameters were obtained, including fractional area change, tricuspid annular plane systolic excursion(TAPSE), myocardial performance index, tricuspid lateral annular systolic velocity(Tricuspid s′). Two-dimensional (2D) RVFWLS was calculated by two-dimensional speckle tracking echocardiography (2D-STE). Right ventricular (RV) end-diastolic volume (RVEDV), RV end-systolic volume (RVESV), RV stroke volume (RVSV), RV ejection fraction (RVEF) and 3D-RVFWLS were measured by 3D-STE. The degree of MF was quantified using Masson′s trichrome stain in RV myocardial samples after heart transplantation. Patients were divided into mild, moderate, and severe groups according to the degree of MF on histology, then echocardiographic parameters were compared among the 3 groups. Pearson correlation analysis and the multiple linear regression analysis between echocardiographic parameters and RV MF were analyzed.Results:Compared with patients with mild and moderate MF, 3D-RVFWLS, 2D-RVFWLS and conventional parameters of RV function were significantly decreased in patients with severe MF.RV MF strongly correlated with 3D-RVFWLS ( r=-0.71, P<0.01), modestly correlated with 2D-RVFWLS ( r=-0.53, P<0.01), and weakly correlated with RVFAC, TAPSE, RVEF, Tricuspid s′, RVSV ( r=-0.47, -0.44, -0.35, -0.29, -0.38; all P<0.01). 3D-RVFWLS correlated best with the degree of MF compared with 2D-RVFWLS and conventional RV function parameters ( r=-0.71 vs r=-0.29~-0.53, all P<0.05). A stepwise multivariate analysis showed that 3D-RVFWLS was independently associated with RV MF (β=1.554, P<0.01, adjusted R2=0.539). Conclusions:3D-RVFWLS can provide an important imaging reference for detecting the degree of RV MF in patients with end-stage HF.

Objective To investigate the effect of endovascular treatment in iliac femoral artery occlusive disease of TransAtlantic Inter-Society Consensus (TASC) C,D lesions in elderly patients.Methods Totally 104 elderly TASC C,D patients with iliac femoral artery occlusion were treated,including TASC C (n =73),TASC D (n =31).Occlusive length range were 4.25-17.61 cm,with an average of (13.46±1.24)cm.The curative effect,complications and following up after surgery were analyzed.Results Ninty-four patients were treated successfully,and the technical success rate was 90.38％ (94/104);12 patients underwent balloon dilation,92 patients underwent balloon dilation combined stent.There were 8 cases (8/104,7.69 ％) with periprocedural complications,included arterial thrombosis (n =2),pseudoaneurysm (n=1),heart failure (n=4),and radiographic contrast nephropathy (n=1).Doppler ankle-brachial index increased from 0.32±0.12 to 0.81±0.11 (P＜0.01) at 1 week after treatment.The patency rates were 88.46 ％ (69/78) and 81.13％ (43/53) at 6 and 12 months followed-up respectively.Conclusion Minimally invasive endovascular treatment has high technical success rate,low complication rate,and satisfactory clinical curative effect for iliac femoral artery occlusive disease of TASC C,D lesions in elderly patients.

Acute Coronary Syndrome ; complications ; drug therapy ; Aged ; Hematoma ; complications ; Heparin, Low-Molecular-Weight ; adverse effects ; therapeutic use ; Humans ; Leg ; Male ; Thrombolytic Therapy ; adverse effects

Two main impact factors on accuracy of electronic balance are discussed based on its working principle:acceleration of gravity and temperature variation.Through analysis of interacting variables in acceleration of gravity and electronic balance measurement results,the method of builtin standard weight compensation and its shortages are brought forward and attach the compensation method on weight calibration.The impacts on temperature change for the electronic balance measurement are analyzed,which proposes a method to eliminate these factors.

Objective To determine the plasma and intracellular concentration of Ara-C by the RPHPLC method and analyse the influence factors and the relationship between the concentration and drug dose.Methods Mononuclear cells and serum of 75 patients with acute leukemia were extracted after the first intravenous infusion of different administration dosage of Ara-C (0.5, 1.0, 2.0 g/m2), and analysed with different chromatographic conditions by RP-HPLC. Results The linear range of Ara-CTP was 0.28-18.96 μg/ml (r =0.998), and the detection limit was 0.28 μg/ml. The detection limit of Ara-C and Ara-U in plasma was 0.0157 μg/ml and 1.034 μg/mnl respectively. In 27 samples preserved for more than 1.5 years, 11 (40.7 %)cases of the plasma concentration of Ara-C were below the detection limit. In 36 samples of mononuclear cell count below 1.5×106/ml, 15 cases (41.7 %) of intracellular concentration of Ara-CTP were below the detection limit. The plasma concentration of Ara-U and intracellular concentration of Ara-CTP were increased with administration dosage of Ara-C increased, and the plasma concentration of Ara-C was not increased. The intracellular concentration of Ara-CTP in old patients over 40 years was tend to in crease with age.Conclusion The RP-HPLC method is simple, rapid, stable, reproducible and applicable for the monitoring of the plasma concentration of Ara-C and intracellular concentration of Ara-CTP. In 0.5-2.0 g/m2 dose range of Ara-C, the plasma concentration of Ara-U and intracellular concentration of Ara-CTP was increased with administration dosage of Ara-C increased.