To establish an HPLC-MS/MS method for the analysis of quercetin, kaempferid and isorhamnetin in rats plasma and study its pharmamacokinetics after an intragastrical administration of Hippophae rhamnoides extracts. Five healthy male Sprague-Dawley (SD) rats were given single doses of H. rhamnoides extracts (quercetin 26.35 mg x kg(-1), kaempferid 4.040 mg x kg(-1), isorhamnetin 31.37 mg x kg(-1)), and then their orbital sinus blood samples were collected at different time points. The drug plasma concentration of the three flavonoids was determined by HPLC-MS/MS method. After that, the main pharmacokinetics parameters were calculated by using Kinetica 5. 0. 11 software. The methodological test showed that the linear concentration ranges of quercetin, kaempferid and isorhamnetin were 7.500-600.0 μg x L(-1) (R2 = 0.998 5), 1.000-80.00 μg x L(-1) (R2 = 0.998 5 ) and 10.00-800.0 μg x L(-1) (R2 = 0.998 0), respectively. The inner and inter-days precisions were both less than 14.0%. The plasma samples showed a good stability and consistency with the requirement of biological sample analysis after the samples were frozen once and placed at - 20 degrees C for 15 d and room temperature for 6 h and the treated analytes were placed at -20 degrees C for 24 h. For quercetin, the pharmacokinetic parameter t(½β), AUC(0-∞), MRT(0.∞), C.(max) and T(max) were (113.3 ± 19.37) min, (12 542.14 ± 3 504.05) μg x h x L(-1), (119.6 ± 13.29) h, (164.6 ± 27.33) μg x L(-1) and (5.199 ± 0.840 3) h, respectively. For kaempferid, the pharmacokinetic parameters t(½β), AUC(0-t), MRT(0-∞), C(max) and T(max) were (79.85 ± 17.15) min, (934.51 ± 94.59) μg x h x L(-1), (81.50 ± 13.75) h, (80.15 ± 14.24) μg x L(-1) and (3.827 ± 0.902 7) h, respectively. For isorhamnetin, the pharmacokinetic parameters t1,2,, AUC(0-t), MRT(0-∞), C(max) and T(max) were (118.3 ± 20.73) min, (26 067.77 ± 4 124.60) μg x h x L(-1), (129.0 ± 16.30) h, (269.6 ± 29.32) μg x L(-1) and (6.513 ± 1.450) h, respectively. The HPLC-MS/MS analysis method established in this study was proved to be sensitive and accurate and could be applied in the pharmacokinetic study of quercetin, kaempferid and isorhamnetin in rat plasma.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Hippophae ; chemistry ; Kaempferols ; blood ; pharmacokinetics ; Male ; Quercetin ; analogs & derivatives ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

This paper presents a new kind of collecting and processing system of the sound signal of larynx where double sound cards and software filter are used. The installation of the double sound cards and processing proposal of software are mainly discussed and a new kind of ADC method of dual-channel sound signal is put forward in this paper. This system has the feature of reliable performance, simple installation and easy maintenance.
Algorithms ; Equipment Design ; Humans ; Larynx ; physiology ; Microcomputers ; Signal Processing, Computer-Assisted ; instrumentation ; Software ; Sound ; Speech Production Measurement ; instrumentation

Objective To evaluate the effectiveness and safety of a sodium hyaluronate-containing dressing for skin wound healing after Q-switched laser therapy.Methods Sixty-six patients with facial pigmentary disorders were enrolled in this study.After treatment with Q-switched laser,the patients were randomly and equally divided into three groups to apply a sodium hyaluronate-containing dressing,a thermal spring water-containing facial mask and a distilled water-containing facial mask respectively.The dressing or facial masks were externally applied to the face for 15 minutes once immediately after the laser therapy,then,once every night for 14 consecutive days.Inflammatory responses,such as erythema,edema and burning sensation,were measured semi-quantitatively on day 1,3,7 and 14 separately after the laser therapy.The time for skin wound healing and degree of patients' satisfaction were compared among these groups by chi-square test and rank sum test,respectively.Results The sodium hyaluronate-containing dressing and thermal spring watercontaing facial mask were superior to the distilled water-containing facial mask in the improvement of erythema,edema and burning sensation after the laser therapy (all P ＜ 0.05).Compared with the patients applying the thermal spring water-containg facial mask and those applying the distilled water-containing facial mask,those applying the sodium hyaluronate-containing dressing felt better,with significantly increased skin cleanliness,humidity,smoothness,confort degree and overall satisfaction (all P ＜ 0.05).Conclusion The sodium hyaluronate-containing dressing is effective for improving acute inflammatory responses and increasing skin cleanliness,humidity and smoothness after laser therapy.

Since the polyjuice potion ingredient is complex, we need to develop an analysis method with well separation and high stability to perform qualitative analysis. After dividing chemical components of Mahuang Decoction into fat-soluble and water-soluble constituents by gradient extraction, GC-MS was used to analyze the chemical components of the ethyl acetate extraction. The results showed that forty compounds had been identified by NIST MS search 2.0 standard mass spectrometry Library and literatures. Next, UPLC-Q-TOF-MS was applied to idendify the chemical components of the water extraction. The results showed that thirty-nine compounds had been identified by MZmine-2.9.1, Isotope Pattern, fragmentation regularity of mass spectrometry and literatures. This experiment will provide evidences for elucidation of the effective substance in Mahuang decoction and can be used as a simple, shortcut method for analysis and identification for the polyjuice potion.
Chromatography, High Pressure Liquid ; methods ; Ephedra ; chemistry ; Ephedra sinica ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods

Background Cellular autophagy is a non-apoptosis death form of tumor tissue.Research determined that arsenie trioxide (As2O3) leads to apoptosis of tumor cells.But whether As2O3 induce autophagy of SO-Rb50 cells or not is unclear.Objective This study was to assess the effects of As2O3 on autophagy of SO-Rb50 cells.Methods As2O3 with the concentration of 0,0.5,1.0,2.0,4.0 μmol/L was used to treat the SO-Rb50 cell line for 48 hours,and the growth and proliferation of SO-Rb50 cells were detected using MTT assay (A570).pGFP-LC3,a marker of autophagy,was constructed to transfer SO-Rb50 cells,and the cells were then divided into RPMI-1640 culture group (untreated group),As2O3 + RPMI-1640 culture group (As2O3 treated group) and rapamycin culture group (positive control group).Autophagy of SO-Rb50 cells was examined by laser confocal microscope and monodansylcadaverine (MDC) influorescence staining,respectively,48 hours following cell culture.Ultrastructural features of autophagy were examined with transmission electron microscope (TEM).The percentage of autophagy positive cells in different concentrations of As2O3 treated groups was calculated with flow cytometer.Results The A570 values of SO-Rb50 cells were 2.194±0.066,1.841 ±0.213,1.035±0.046,0.374±0.042 and 0.167±0.019 in 0,0.5,1.0,2.0,4.0 μmol/L As2O3 treated groups,with a significant difference among these 5 groups(F=547.636,P＜0.05),and those of 0.5,1.0,2.0,4.0 μmol/L As2O3 treated groups were significantly reduced in comparison with untreated group (P =0.000).The positive granular spots for GFP-LC3 chimeric protein were seen to aggregate in autophagic vacuoles in the As2O3 treated group and positive control group,but diffuse cytoplasmic signal for GFP-LC3 was found in the untreated group.Normal ultrastructure of SO-Rb50 cells was exhibited in the untreated group,and many double-membrane-like bound vesicles and autlysosomes were documented in the As2O3 treated group and positive control group under the TEM.A lots of MDC fluorescence granule were found in the As2O3 treated group and positive control group rather than the untreated group.Flow cytometry showed that the percentages of SO-Rb50 cells were 0,15.6％,42.7％,57.9％,79.5％ and 89.0％ in the 0,0.5,1.0,2.0,4.0 μmol/L As2O3 groups and positive control group,respectively,showing a As2O3 concentration-dependent increase.Conclusions As2O3 can induce the autophagy of SO-Rb50 cells and inhibit the proliferation of SO-Rb50 cells.Autophagic response of SO-Rb50 cells appears prior to the nuclear change after exposed to As2O3.The degree of autophagy of SO-Rb50 cells is associated with As2O3 dose.

Objective To apply a simple fixator to salivary gland scintigraphy to evaluate its effect on body position movement. Methods Totally 30 female patients complaining xerostomia were randomly and equally divided into a conventional scan group and a simple fixator group,who were injected with Technetium Tc-99m Pertechnetate intravenously. In the conventional scan group the patients had their necks fixed with the bracket and cushion,while in the other group the fixation was executed with the single fixator. SPECT imager was used for dynamic salivary gland scintigraphy, and two nuclear medicine physicians evaluated the maximum shift of the parotid gland along left-right(X)and head-foot(Y)axes in series of dynamic images. SPSS 16.0 software was used for data analysis. Results In the conventional scan group the maximum shift was(6.6±4.6)mm at X axis and(5.2±3.4)mm at Y axis;in the single fixator group the maximum shift was(3.2± 1.6)mm at X axis and(3.0±1.3)mm at Y axis.There were significant differences between the maximum shifts in the two groups (P=0.012,X axis;P=0.027,Y axis).Conclusion The single fixator assists in salivary gland scintigraphy,and alleviates the body position movement during dynamic acquisition and provides data support for functional parameter calculation and result determination.

Objective To investigate the curative effect of brucellar spondylitis,so as to provide scientific proof for improving the curative level of the disease.Methods Epidemiological information was collected from 113 patients diagnosed as brucellar spondylitis,who were divided into 5 groups according to different drugs and drug combinations of doxycycline,gentamicin,sulfamethoxazole and rifampicin.Then the curative effect was investigated.Twenty-one patients who had greater paoas muscle abscess or Para vertebral abscess,intraspinal abscess of spinal canal,necrotic intervertebral disk and major osteolasia received the minimal invasive surgery and the focus removal surgery.Results The occurrence of the disease in female was much higher than that in male.Grazing and breeding beasts was the principal route of infection.Lumbars was mostly involved.they usually was infected in the adjacent 2 piece.L4 was the most common and seriuous one.The curative effect of doxycycline group was better than that without doxycycline(72.60% vs 35.00%,X2=15.14,P＜0.05).Doxycyeline+gentamicin+sulfamethoxazole was reeommended as the first choice.However,the curative effect did not increase despi~course of the treatment prolonged.The heMing rate and effective rate after 1 course was 52.21%(59/113)and 92.04%(104/113).that after 2 courses 58.41%(66/113)and 95.58%(108/113),that after 3 courses 59.29%(67/113)and 95.58%(108/113).The healing rate in different course did not present differences(P＞0.05).21 patients undergoing surgery were followed-up,12 patients were after 2 years and 9 patients were between 1-2 years.The healing rate was 95.24%(20/21),1 case was healed basically,the effective rate was 100%.None reoccurred.Conclusions There are characterized features in clinical epidemiology of the brucell spondylitis.Long term,adequate in dosage,combination and multi-approach use of antibiotics is the most reliable way to treat and prevent it from recurring.But fof the patients soitable for surgery.the minimal invasive surgery or the focus removal could shorten the course of therapy,decrease the complications and increase the cure rate.

Objective To study the relationship between environmental chemical elements,vires infection and dilated cardiomyopathy(DCM).Methods In 2008,233 patients with DCM(case group)and 150 patient with stable angina(control group)were chosen in Liaocheng People's Hospital and Yanggu People's Hospital,Shandong province.Population distribution and disease history were surveyed in the two groups.Human myocardial antibody IgG(AMA-IgG),Coxsackie B virus IgG(CBV-IgG),Adenofirus antibody IgG(ADV-IgG)were detected by ELISA in both the case group and the control group.Serum trace elements were detected in the two groups.The general chemical and toxicological indicators in drinking water of the high-and the low-incidence aireas of the disease were control group[60.00%(90/150),χ2=13.80,P<0.01)].Per capita annual income(Yuan,RMB)in the case group (3207.82±618.51)was lower than that of the control group[(5086.61±886.12),t=24.40,P<0.01].Personal alcohol consumption in the case group[(175.00±160.50)g/d]was higher than that of the control group[(110.22±100.03)g/d,t=4.40,P<0.01)].The rate of myocarditis in the case group[5.15%(12/233)]was higher than ADV-IgG in the cage group were 7.78%(7/90),6.67%(6/90)and 6.67%(6/90),respectively.Compared with those in the control group[3.33%(2/60),5.00%(3/60)and 5.00%(3/60),χ2=1.26,0.18,0.18,all P>0.05],no mg/L]in drinking water of the high-incidence areas were significantly higher than that of iron[(0.39±0.67)mg/L,t=2.11,P<0.05]and that of manganese[(0.15±0.14)mg/L,t=3.01,P<0.01]in the low-incidence arefas.The content of semm iron[(69.1±57.8)μmol/L]in the case group evidently exceeded the normal range(15.6-35.9 μmol/L)and obviously higher than that in the control group[(20.0±17.5)μmol/L,t=5.04,P<0.01].Conclusions Theso data do not support that DCM is related with persistent virus infection and autoimmunization.DCM is probably related with low incomes,high alcohol consumption,myocarditis,high iron and manganese contents in drinking water and high content of serum iron.

Aim To study the inhibitory effect of volatile components in Oroxyli Semen on liver cancer and its possible mechanisms.Methods H22 bearing mouse model was used,the mice were divided into six groups:blank,model,positive (cytoxan,100 mg · kg-1),low-,mid-,and high-dose (17.5,35,and 70 mg · kg-1) volatile components groups,and then the mice were ig given once daily for consecutive 12 d.Then the tumor growth inhibitory rate,spleen and thymus indexes were calculated;the serum levels of IL-2,IL-6 were determined.HE staining was used to study of the apoptosis of the solid tumor.After treatment of SMMC-7721 cells with 0 ～ 1 g · L-1 of volatile components for 24,48 and 72 h,MTT assay was used to examine the proliferation.TUNEL method was applied to detect cell apoptosis,and RT-PCR method to detect Bax,Bcl-2,caspase-3 mRNA experssion.Results The inhibitory rate of volatile components high-dose on H22 bearing mice was 42.08％.The thymus index and the contents of serum IL-2 and IL-6 of H22 bearing mice were significantly higher than those in model group.Volatile components could significantly inhibit proliferation and induce apoptosis of SMMC-7721 cells,downregulate the expression of Bcl-2 mRNA,and up-regulate the expression of Bax,caspase-3 mRNA.Conclusions The volatile components in Oroxyli Semen have obvious anti-tumor activity in vitro and in vivo,and its mechanism may be related to enhancing immune system and promoting tumor cell apoptosis.

OBJECTIVETo confirm whether human MHC class I chain-related A (MICA) induces the amplification of V delta 1 gamma delta tumor-infiltrating lymphocytes (TILs) in vitro and to identify the cytotoxicity of MICA-reactive V delta 1 gamma delta TILs towards epithelial tumor cells.

METHODSMICA protein was prokaryoticly expressed and purified by molecular cloning technology. The purified recombined MICA (rMICA) was used to induce V delta 1 gamma delta T cells from tumor tissues in vitro and the cytotoxicity of these V delta 1 gamma delta TILs were tested by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT).

RESULTSThe rMICA was expressed in prokaryocyte with pET30 as a vector. The immobilized rMICA protein could markedly induce the amplification of V delta 1 gamma delta T cells from tumor tissue in vitro. These V delta 1 gamma delta T cells showed strong cytolytic activities towards tumor cell lines expressing MICA.

CONCLUSIONThe MICA-reactive V delta 1 gamma delta T cell may be a candidate for adoptive cellular therapy of tumors.

Adult ; Aged ; Female ; HeLa Cells ; pathology ; Histocompatibility Antigens Class I ; biosynthesis ; genetics ; Humans ; Immunotherapy, Adoptive ; Lymphocytes, Tumor-Infiltrating ; immunology ; Male ; Membrane Proteins ; biosynthesis ; genetics ; Middle Aged ; Ovarian Neoplasms ; immunology ; Receptors, Antigen, T-Cell, gamma-delta ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; T-Lymphocytes, Cytotoxic