Objective: To optimize the extraction process of effective components in Lycii Fructus using central composite design(CCD)-response surface method(RSM). Methods: The yield of main effective components, polysaccharides, flavonoids, rutin and chlorogenic acid as well as the yield of total extract were used as indicator for evaluating the effect of extraction solvent, extraction time, solid- liquid ratio, extraction temperature and other factors. The single factor experiment was conducted to determine the range of various factor parameters, the factor design experiment was performed for the screening of the factors that had significant influence on the extraction efficiency of effective components, the maximum velocity rise experiment was carried out to determine the central value of the factors, and finally the optimal extraction process was optimized by the central composite design- response surface methodology. Results: The optimum extraction conditions of Lycii Fructus were obtained as follows: the extraction with 60% aqueous ethanol solution by ultrasound sonication at room temperature for 40.5 minutes, and the solid- liquid ratio was 1:27. Conclusion: The present optimal extraction process could effectively improve the extraction efficacy of effective components from Lycii Fructus, and the created CCD-RSM model and related conditions have good predictability and repeatability, which provides experimental and theoretical reference for further development and research of Lycii Fructus.

BACKGROUND:As the multipotent differentiation potential, bone marrow mesenchymal stem cells exert an important role in bone metabolism disorders, which is regulated by a variety of hormones and cytokines. Currently, the epigenetic regulatory mechanisms underlying osteogenic differentiation of bone marrow mesenchymal stem cells are unclear, and association between histone deacetylase (Hdac) and osteoporosis needs to be further explored. OBJECTIVE:To investigate the epigenetic mechanisms of bone formation by analyzing the expression of Hdac1, 3, 4 mRNA profile in bone marrow mesenchymal stem cells isolated from ovariectomized mice. METHODS:A total 30 female healthy Kunming mice were randomly divided into sham group and ovariectomy group. After 7 days of adaptive feeding, mice in the ovariectomized group (n=15) were subject to bilateral ovariectomy;mice in sham group (n=15) were sham-operated. RESULTS AND CONCLUSION:In the ovariectomy group, the trabecular bone of the femur was sparse or broken, the width of trabecular bone was narrowed, the trabecular separation was widened, and the trabecular bone volume was reduced. The level of Hdac3 mRNA was lower in bone marrow mesenchymal stem cells from ovariectomized mice compared with controls, but there was no significance between Hdac1, Hdac4 mRNA expression of the two groups. These findings demonstrate that Hdac might play an important role in bone remodeling in the model of estrogen deficiency-induced osteoporosis.

Background Researches showed that as the non-optical factors,cognitive has certain influence on the regulating system.So accurately experimental design is one of the key steps that evaluates the non-optical factors on regulating system.Objective The present study was to investigate the influence of presenting pattern of target and watching way on the level and amplitude of accommodative fluctuate and to analyze the effect of focus gaze of cognitive on regulating system and the relationship between focus gaze condition under near work and the development of myopia.Methods This study complied with Declaration of Helsinki,and the permission of Ethic Committee and written informed consent was obtained before entering in this trial.Thirty healthy volunteers were included with the mean age (24.80 ± 1.98) years old,equivalent refractive diopter (-1.92 ± 2.02) D and mean cylinder (-0.19±0.58) D.The presenting pattern of the targets was designed as focus gaze and relaxed gaze.The accommodative response and accommodative fluctuation in the complete corrected right eyes for the different targets at the 40 cm under the gazing state was recorded with Grand Seiko WAM 5500 automatic infrared refractor in the experiment.Results The mean accommodative response value was (1.86±0.26) D under the focus gaze and (1.27±0.39) D under the relax gaze,showing a statistically significant difference (t=-8.052,P=0.000).The mean fluctuate value was(0.17±0.06) D under the focus gaze,with a significant lowing in comparison with (0.28±0.17) D under the relax gaze (t =3.600,P =0.001).Conclusions These results demonstrate that the different presenting patterns of sighting target and watching ways of the subjects affect accommodation system.The accommodative response was relatively more accurate with a smaller microwavc moving under the focus gaze condition.

Background and purpose:Chemotherapy plays an important role in the treatment of breast carcinoma by inhibiting the tumor growth and inducing the apoptosis.MAPK transduction pathway is closely related to proliferation and apoptosis of varieties of tumor cells,inhibition of MAPK pathway may increase the efficiency and decrease the toxicity of chemotherapy.Our study was to investigate the effect of MEK inhibitor PD98059 in response of breast cancer cell lines to Epirubicin.Methods:Human breast cancer cell lines MCF-7 and MCF-7/ADR were used as cell models.Epirubicin(EADM),PD98059(inhibitor of MAPK Kinase-MEK),or EADM+PD98059 was added into the culture medium,the expression of MEK2 and p-ERK were measured by Western blot,the growth of the two cell lines were measured by MTT.Results:ERK activity was elevated in MCF-7 after the treatment of EADM,the cells were more sensitive to EADM if combined with PD98059,while in MCF-7/ADR,ERK activity kept unchanged after EADM treatment,and PD98059 has no effect on the sensitivity of cells to EADM.Conclusion:MAPK signal transduction may be activated in some cells treated by EADM,adding inhibitor of MAPK signal transduction could improve the sensitivity of the cells to EADM.

DNA microarrays have been acknowledged to represent a promising approach for the detection of viral pathogens. However, the probes designed for current arrays could cover only part of the given viral variants, that could result in false-negative or ambiguous data. If all the variants are to be covered, the requirement for more probes would render much higher spot density and thus higher cost of the arrays. Here we have developed a new strategy for oligonucleotide probe design. Using type I human immunodeficiency virus (HIV-1) tat gene as an example, we designed the array probes and validated the optimized parameters in silico. Results show that the oligo number is significantly reduced comparing with the existing methods, while specificity and hybridization efficiency remain intact. The adoption of this method in reducing the oligo numbers could increase the detection capacity for DNA microarrays, and would significantly lower the manufacturing cost for making array chips.

Objective To appraise and compare protein expression profiles in sera of patients without or with recurrence following liver transplantation for hepatocellular carcinoma (HCC) using SELDI-TOF-MS technique,and establish the diagnostic and predictive model. Methods A total of 76 sera (41 from disease free survival patients and 35 from recurrence individuals) were collected pretransplantation and differentially expressed proteins were identified by SELDI-TOF-MS. The intensity values for each peak were analyzed by Biomarker Wizard Software to screen serum proteome biomarkers related to the recurrence post-transplantation. By using Biomarker Patterns Software, the classification trees were generate. from randomly selected samples (30 fingerprints obtained from each group). The sensitivity and specificity of best decision tree were then chosen for blind test with 16 samples (5 from recurrence individuals and 11 from recurrence-free survival patients). Results There were significant differences only in tumor size and the presence of vascular invasion between recurrence group and recurrence-free survival group (P＜0.05). According to serum protein fingerprints, a total of 368 protein peaks were identified at the mass-to-charge ratio (M/Z) value ranging from 2000 to 300 00. There were 22 significant differential proteins between two groups. Among them, 9 proteins were up-regulated and 13 proteins were down-regulated -espectively in recurrence group. The intensity values of differential proteins were input into BPS for classification tree analysis and the best performing tree could distinguish two groups successfully. As a result of blind assessment for this model,a sensitivity of 80.0 % (4/5) and specificity of 72.7 % (8/11) were obtained. Conclusion Some of differential proteins screened by SELDI-TOF-MS technique in the serum may be correlated with the prognoses of liver transplantation patients with HCC. The decision tree may be useful for the clinical application of formulating the indication for liver transplantation, detecting extrahepatic micrometastasis and setting up the diagnostic and treatment strategies.

Objective To investigate the better regimen of combined cyclophosphamide pulse therapy with corticosteroids in the treatment of Pemphigus. Methods Intravenous cyclophosphamide was given weekly in a dosage of 600 mg to those Pemphigus patients whose conditions couldn't be improved by corticosteroids (oral prednisone 1.2～1.5 mg/kg/d) alone. Results Ten patients with Pemphigus received weekly cyclophosphomide therapy in addition to corticosteroid. Patients conditions improved quickly, without side effects. Conclusions Cyclophosphamide weekly pulse therapy combined with corticosteroids is a good regimen in the treatment of Pemphigus.

OBJECTIVETo establish a method for real-time recording the oxygen consumption of mice under normobaric hypoxia.

METHODSThe experimental apparatus was made up of animal container, filling water control system, electronic balance, hose, a computer with weight recording software, etc. The working principle was that the oxygen consumed by animal was replaced by water filling which was controlled by the pneumatic and hydraulic actuator. The water was weighted by an electronic balance and the weight signal was recorded into excel file at the same time. The accuracy and precision of the apparatus were detected by a 10 ml syringe. The oxygen consumption characteristics of 6 acute repetitive hypoxia mice and 6 normal mice were observed.

RESULTSThe P value for the paired t test was 1 and the CV value was 4%. The survival time and total oxygen consumption of acute repetitive hypoxia mice were both significantly increased compared to normal mice (P < 0.05), which were (58.8 +/- 6.8) min and (46.0 +/- 8.7) min respectively for the survival time and (85.1 +/- 8.5) ml and (73.6 +/- 5.4) ml respectively for total oxygen consumption.

CONCLUSIONThe hypoxia tolerance of the acute repetitive hypoxia mice can significantly improved by taking more oxygen in the animal cabin. The accuracy and precision of the apparatus are high and it can be used for the determination of oxygen consumption in hypoxia research.

Animals ; Antibodies, Monoclonal ; immunology ; Atrazine ; analysis ; immunology ; Chromatography, Gas ; Chromatography, High Pressure Liquid ; Enzyme-Linked Immunosorbent Assay ; Food Contamination ; analysis ; Herbicides ; analysis ; immunology ; Mass Spectrometry

This study is to investigate the protection effect of schisandrin B (Sch B) against oxidation stress of HK-2 cells induced by cisplatin and the mechanisms involved. HK-2 cells were cultured and divided into different groups: solvent control group, cisplatin exposure group, positive group, Sch B treatment group. Cell viability and toxicity were evaluated by MTT and LDH assay. GSH level and SOD enzymes activities were also measured. DCFH-DA as fluorescence probe was used to detect ROS level by fluorescence microplate reader. Nrf2 translocation was detected by Western blotting. Real time Q-PCR was used to detect expressions of NQO1, HO-1 and GCLC mRNA level. The results showed that Sch B could significantly inhibit the decline of cell viability induced by cisplatin treatment (P < 0.05) and the protective effect was in a dose dependent manner. Furthermore, Sch B treatment significantly inhibited the increase of ROS level induced by cisplatin and reversed the decrease of GSH level (P < 0.05). When Sch B concentration was up to 5 micromol x L(-1), SOD enzyme activities were also enhanced significantly compared with that of the cisplatin group (P < 0.05). It was shown that Sch B could cause nuclear accumulation of Nrf2 in association with downstream activation of Nrf2 mediated oxidative response genes such as GCLC, NQO1 and HO-1. These results suggested Sch B could protect against the oxidative damage of HK-2 cells induced by cisplatin via the activation of Nrf2/ARE signal pathway.
Antineoplastic Agents ; toxicity ; Antioxidants ; isolation & purification ; pharmacology ; Cell Line ; Cell Survival ; drug effects ; Cisplatin ; toxicity ; Cyclooctanes ; isolation & purification ; pharmacology ; Glutamate-Cysteine Ligase ; genetics ; metabolism ; Glutathione ; metabolism ; Heme Oxygenase-1 ; genetics ; metabolism ; Humans ; Kidney Tubules, Proximal ; cytology ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Lignans ; isolation & purification ; pharmacology ; NAD(P)H Dehydrogenase (Quinone) ; genetics ; metabolism ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Polycyclic Compounds ; isolation & purification ; pharmacology ; RNA, Messenger ; metabolism ; Reactive Oxygen Species ; metabolism ; Schisandra ; chemistry ; Signal Transduction ; Superoxide Dismutase ; metabolism