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 evaluate the effects of valsartan and angiotensin-converting enzyme inhibitors on the excretion rate of urinary albumm(UAER) and the clearance of endogenous creatinine(Cr-C) in elderly patients.Methods 120 elderly patients with stage of the 2～3 class hypertension were included.The patients were divided into 3 groups,the valsartan group and the benazpril group and captopril group,the anti-hypertensive effects and UAER and Cr-C were determined before and after the treatment.Results No significantly different anti-hypertensive effects were observed in valsartan,benazpril and captopril.But the effects of UAER,Cr-C in valsartan group had significant changes compared with the other groups,and the three groups had significant differentiation(P

High mobility group protein B1 (HMGB1) is the most representative substance in the alarmins family, it is actively or passively release to extracellular by the activation of monocyte/macrophage and the dead cells, and then it stimulates the production of a variety of inflammatory mediators, and increases the organism's inflammatory response through relevant receptors signaling pathways. In recent years, its concentration can reflect the severity of inflammation and injury and was related to the prognosis, HMGB1 has won more and more attention in the development of sepsis. By reviewing the study of HMGB1 in sepsis pathogenesis, signal pathway and reversal measures, it was found that HMGB1 was considered as an important inflammatory mediators and warning signal involved in the pathogenesis of sepsis, and was become a new target in the treatment of sepsis. Further research on the role of HMGB1 in the pathogenesis of sepsis is needed in the future, and the development of new drugs combined with HMGB1 will be used in the study of HMGB1 in animal experiments.

Objective To investigate the protective effect of agmatine (AGM) against peritoneal inflammatory response and neutrophil (PMN) infiltration induced by zymosan (ZYM) in mice. Methods Thirty-six adult male C57BL/6 mice were randomly divided into sham group, model group, and AGM treatment group. Peritonitis model was reproduced by intra-peritoneal injection of 1 mg/mL ZYM (0.5 mL), while equivalent phosphate buffer saline (PBS) was given to sham group. 200 mg/kg AGM was injected into peritoneal cavity after ZYM challenge in AGM treatment group. Six mice in each group were sacrificed at 2 hours and 6 hours, respectively, after reproduction of the model. Blood sample and peritoneal lavage fluid (PLF) were collected. The levels of keratinocyte-derived chemokine (KC), macrophage inflammatory protein 2 (MIP-2), tumor necrosis factor-α (TNF-α), interleukins-6 (IL-6) in serum and PLF were determined by enzyme linked immunosorbent assay (ELISA). The number of leukocytes and PMN in PLF were determined by hemocytometer and flow cytometry, respectively. Results Compared with sham group, all serum and PLF levels of KC, MIP-2, TNF-α and IL-6 were greatly elevated at 2 hours after ZYM injection in model group, while AGM treatment could dramatically reduce the levels of the above-mentioned cytokines in serum and PLF as compared with those of the model group [serum KC (ng/L): 990.7±137.9 vs. 2 053.2±262.7, MIP-2 (ng/L): 642.2±124.4 vs. 1 369.7±146.5, TNF-α (ng/L): 608.6±38.1 vs. 1 044.7±101.0, IL-6 (ng/L): 1 058.2±129.1 vs. 1 443.3±190.1; PLF KC (ng/L): 7 462.3±839.6 vs. 12 723.5±1 515.7, MIP-2 (ng/L): 1 570.8±193.4 vs. 3 471.4±384.7, TNF-α (ng/L): 1 115.8±156.7 vs. 1 499.2±231.2, IL-6 (ng/L): 2 646.5±223.2 vs. 3 126.7±291.4; all P < 0.05]. The expressions of KC, MIP-2 and TNF-α at 6 hours were significantly lower than those at 2 hours in model group and AGM treatment group, but IL-6 levels were further increased. The levels of KC and MIP-2 in serum and PLF at 6 hours were decreased to the levels of sham group. At 6 hours after the reproduction of the model, the number of total inflammatory cells and PMN of PLF in the model group was significantly higher than those of the sham group. In contrast, AGM notably lowered the number of inflammatory cells and PMN in peritoneal fluid after ZYM attack [total inflammatory cells (×109/L): 14.7±1.1 vs. 2.0±0.4, 10.1±1.2 vs. 14.7±1.1; PMN (×109/L): 11.37±1.22 vs. 0.18±0.05, 7.69±0.57 vs. 11.37±1.22, all P < 0.05]. Conclusion AGM can effectively alleviate acute peritoneal inflammatory injury induced by ZYM, mainly through reducing the secretion of inflammatory mediators and chemokines, and inhibiting the infiltration of leukocytes and neutrophils.

Objective To develop MSNs loaded with ICG and investigate its diagnostic value and photodynamic therapeutic value in the experimental pancreatic cancer. Methods ICG was loaded into MSNs to prepare the ICG/MSNs probe. The probe toxicity was evaluated by MTT assays. Near?infrared stereo fluores?cence microscope ( NISFM) was applied to investigate whether the ICG/MSNs would be uptaken by the human pancreatic cancer cells. After incubated with PBS, ICG (10μg/ml), MSNs or ICG/MSNs (10μg/ml ICG) for 24 h and treated with photodynamic therapy (PDT, (780±25) nm laser, 500 mW/cm2), the human pancreatic cancer cells survival rate was determined by MTT method. The human pancreatic cancer cells were implanted into nude mice to prepare subcutaneous tumor models. The distribution of ICG/MSNs in sub?cutaneous tumor models was studied with in vivo imaging system ( IVIS ) . With reference to the injection dose of ICG(0.5 mg/kg), the mice in PBS group, ICG group, ICG/MSNs group (4 mice per group) were injected via tail vein with 150μl PBS, ICG solution and ICG/MSNs solution, respectively. After treated by PDT for 48 h, the mice were observed by IVIS for 2 weeks using BLI to evaluate the therapeutic effect of PDT. NISFM was used to observe the fluorescence in tumor region. One?way analysis of variance was used to analyze the data. Results The diameter of ICG/MSNs was about 100 nm and it could be uptaken by human pancreatic cancer cells. After treated by PDT, the survival rates of human pancreatic cancer cells were (24?5±5.0)%, (81.2±1.6)%, (90.7±2.0)% and (93.4±1.7)% in ICG/MSNs group, ICG group, MSNs group and PBS group, respectively(F=212.289, P<0.05). ICG/MSNs group showed better therapeutic effect than ICG group( P<0.05) . After 12 d treated by PDT, the tumor did not recur or grow in ICG/MSNs group, but grew obviously in ICG group and PBS group. The BLI of tumor area in PBS group, ICG group and ICG/MSNs group were (61.2±7.7)×108, (56.7±9.0)×108 and (2.4±1.5)×108, respectively(F=67?098, P<0.05) . And the difference between ICG/MSNs and ICG group was statistically significant ( P<0.05) . Meanwhile, the NISFM showed clearly the tumor location with ICG/MSNs. Conclusions ICG/MSNs have good biocompatibility, good PDT effect on pancreatic cancer cells and pancreatic cancer xeno?grafts. Near?infrared fluorescence imaging with ICG/MSNs could delineate the tumor location.

Objective To explore the Chinese medicine syndrome characteristics of mild cognitive impairment (MCI).Methods The papers of MCI syndrome research were reviewed and collected in China National Knowledge Infrastructure,Wan Fang Data and VIP Data from January of 1990 to December of 2014.Statistical analysis was made on the Chinese medicine syndrome types and syndrome factors.SPSS 17.0 software was adopted to make cluster analysis.Combined with experts' experience,related symptoms to the syndrome factors were carried out.Results Totally 32 papers were included.Mter terminology normalization,there were 24 syndrome types of MCI.Top 5 syndromes with high frequency were syndrome of orifices confused by phlegm,syndrome of deficiency of kidney essence,syndrome of deficiency of both Qi and blood,syndrome of internal exuberance of heat toxin and syndrome of blood stasis blocking brain.In syndrome factors of disease location type,kidney and brain covered the highest proportion,30.83％ and 30.00％.In syndrome factors of disease cause and character types,Qi deficiency covered the most,16.50％.According to results of cluster analysis,combined with experts' experience,15 syndrome factors were extracted,including 69 symptoms.Conclusion Chinese medicine syndrome types of MCI were mainly syndrome of orifices confused by phlegm and syndrome of deficiency of kidney essence.The disease locations were mainly kidney and brain.The disease character was Qi deficiency.

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.

OBJECTIVETo identify the tyrosinase inhibitory constituent quickly from Potentilla bifurca.

METHODThe active constituent was found through fraction collecting and tyrosinase inhibitory activity by bioassay-linked HPLC method.

RESULTThe methanol extracts and BuOH fraction of Potentilla bifurca showed strong tyrosinase inhibitory activities. From BuOH fraction of Potentilla bifurca, the tyrosinase inhibitory constituent was isolated and identified as flavonoid, quercetin-4'-O-beta-D-glucoside. It express stronger tyrosinase inhibition than the known tyrosinase inhibitor, kojic acid (IC50 = 0.28 mmol x L(-1)) with IC50 value of 0.001 9 mmol x L(-1).

CONCLUSIONBioassay-linked HPLC fractionation method was provided for determination the active constituents quickly from herbal medicines.

Enzyme Inhibitors ; chemistry ; isolation & purification ; Kinetics ; Peptides ; chemistry ; isolation & purification ; Plant Extracts ; chemistry ; isolation & purification ; Potentilla ; chemistry

Objective:To explore influencing factors of hospitalization cost for breast cancer inpatients, develop an applicable standard using bundled payment approach, so as to provide reference for health insurance organizations to formulate reasonable imbursement method.Methods:31 792 inpatients′ medical records from 16 tertiary hospitals in Beijing, 2015 were included in the present study. Non-parametric tests and logistic regression model for ordinal categorical variable were applied to explore potential influencing factors. Decision tree model was performed to yield case-mix related groups.Results:The breast cancer inpatients were classified into 17 groups based on the following factors: surgical treatment/non-surgical treatment, surgical modality, radiology treatment, lymphadenectomy, comorbidity. RIV(reduction in variance)was 0.26. CV(coefficient of variation)was 0.24~0.97.Conclusions:It was feasible to apply the case-mix bundled payment method as a reimbursement method for breast cancer inpatients. The present study served as reference for formulating related policies.

Objective: To investigate the feasibility of multiplex real-time RT-PCR with fluorescent probes in early screening of Ph-like acute lymphoblastic leukemia (ALL) and analyze the clinical feature and prognos. Method: A total of 118 adult B-ALL patients diagnosed between October 2010 and March 2016 were enrolled in this study. Multiplex RT-PCR was used to detect the Ph-like ALL related fusion gene and CRLF2 expression in 58 BCR-ABL and MLL rearrangement negative patients. The clinical features, treatment response and prognosis were analyzed in Ph-like fusion gene positive and/or CRLF2 over-expression patients. Result: Among 58 patients, 9 patients (9/58, 15.5%) showed Ph-like ALL related fusion genes positive and 10 patients (10/58, 17.2%) showed CRLF2 over-expression. There were statistical differences in age, WBC count, immunophenotypes, cytogenetics and risk stratification among Ph-like fusion gene positive or CRLF2 over-expression patients, Ph⁺ patients, MLL⁺ patients and B-other patients. The 2-year overall survival rates were 65%, 47%, 64% and 74% respectively among these four groups (P=0.043) . The 2-year relapse free survival rates were 51%, 39%, 62% and 70% respectively among these four groups (P=0.010) . Conclusion Routine screening of Ph-like ALL by multiplex RTPCR is feasible.