AIM: To investigate the regulation of ‘Tiao Gan Fang Yao’(TGFY) on neuroendocrine-immuno-function of bandage-stressed rat . METHODS:The stressed rat model was made by bandage. RIA was adopted to measure the function of hypothalamus-pitutary-adrenal gland axis (HPAA) of stressed rat. Meanwhile, the immunity of stressed rat and the regulation of TGFY were observed.RESULTS:Bandage stress increased the contents of serum corticosterone(CORT), and ACTH, and hypothalamus corticotropin releasing hormone ( P

Objective To observe the effect of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on cholesterol gallstones formation in C57BL/6 mice with diet-induced cholesterol gallstone,and then explore the potential mechanism.Methods Fifty C57BL/6 mice were randomly divided into 5 groups (10 mice in each group),referring to control group,experimental group,experimental plus DHA group,experimental plus EPA group,as well as experimental plus DHA and EPA group.The mice in control group were fed with regular diet,and the rest of the mice with lithogenic diet (LD).Subsequent to feeding the mice with separate diets for two weeks,EPA and/or DHA (70 mg · kg-1 · d-1) were orally administered for eight weeks,while the LD feeding was continued during this period.After a total of 10 weeks,the mice were dissected to observe the gallstone formation.The levels of serum lipids,total cholesterol (TC) and phospholipids (PL) in bile,and TC in the liver were tested,and the protein expression of HMGCR,SRBI,ABCG5/ABCG8,CYP7A1 and ABCB11genes in the liver of mice was measured.Results Compared with the experimental group,the experimental plus EPA group had significantly lower TC in liver (0.033 ±0.008 mmolo/g) and bile (1.807 ±0.381 mmolo/L),and lower relative protein expression levels of HMGCR (0.545±0.098),ABCG5 (0.418±0.089) and ABCG8 (0.501 ±0.151)in liver (P＜ 0.05).The contents of TC in liver and bile,and the protein expression of HMGCR,ABCG5andABCG8 in liver were 0.048 ± 0.006 mmol/g and 2.662 ± 0.339 mmolo/L,and 1.011 ± 0.213,1.037 ± 0.276 and 1.266 ±0.312,respectively.No significant differences were observed between experimental plus DHA group and experimental group (P ＞ 0.05).Conclusions EPA could prevent the cholesterol gallstone formation in mice by decreasing the expression of HMGCR and ABCG5/8 genes in liver,therefore reducing cholesterol synthesis and blocking cholesterol transport from liver to bile as well as diminishing cholesterol content in the bile.However,the inhibition effect of DHA on cholesterol gallstone formation was not obvious.

spectrum beta-lactamase genes:blaOXA-128 and blaOXA-129.

AIM: To study the effect of calcium sensing receptor (CaSR) on icariin (ICA) induced mouse embryonic stem cells ( mESCs) to differentiate into cardiomyocytes in vitro.METHODS:mESCs were cultured to embry-oid bodies ( EBs) by direct suspension method and the differentiation of EBs into cardiomyocytes was induced by ICA.The expression of cardiac specific proteinsα-actinin and cardiac troponin-I ( cTnI) was analyzed by Western blot and immuno-fluorescence.The differentiation rate was determined by flow cytometry.The ultrastructure of the derived cardiomyocytes was further characterized by transmission electron microscopy.The expression of cardiac-specific transcription factors Nkx2.5 and GATA-4,as well as CaSR was detected by Western blot.RESULTS: After induction with ICA, the positive characteristics of myocardial cells appeared in the EBs cultured for 2 d.The expression of cardiac-specific sarcomeric pro-tein actinin (α-actinin) and cTnI showed an overall upward trend by Western blot in different phases of ICA induced differ-entiation.The expression of CaSR, Nkx2.5 and GATA-4 was the highest at an early stage of ICA-induced differentiation. Neomycin (an activator of CaSR) up-regulated CaSR, NKx2.5 and GATA-4 expression in the EBs at early stage of ICA-in-duced differentiation, all of which were reversed by NPS2390 ( an inhibitor of CaSR) .CONCLUSION:CaSR is function-ally expressed in mESC-derived cardiomyocytes, and activation of CaSR is involved in the differentiation of mESCs into car-diomyocytes by facilitating the expression of NKx2.5 and GATA-4.

Objective To evaluate the effect of therapeutic patient education on improving life quality of children with atopic dermatitis (AD). Methods A total of 109 children with AD were enrolled, including 53 patients in the intervention group and 56 patients in the control group. The intervention group was given therapeutic patient education in addition to routine treatment, while the control group was given routine treatment without therapeutic patient education. After three months two groups were compared with the disease severity and quality of life in children and their families. Results Compared with control group, the intervention group had significant improvements in severity of AD (P?=?0.003) and also significant improvements in quality of life (IDQOL and CDLQI) (P?=?0.004). The family life quality (DFI) of the two groups were both improved, but the difference was not signiifcant (P?=?0.492). Conclusions Therapeutic patient education can improve symptoms of atopic dermatitis, and the quality of life of children as well.

Objective To evaluate the use of corrected standard uptake value(SUV)in patient with peripheral solitary pulmonary lesion and the relationship between corrected SUV and the lesion's size.Methods This retrospective study included 69 patients with peripheral solitary pulmonary lesions seen on PET and CT scans whose diagnoses were confirmed by pathology.Each lesion’s FDG uptake was quantified by using two methods:the SUV and the corrected SUV.The lesion’s size was recorded on CT.Results The diagnostic accuracy was 88.7% and 98.5% for PET/CT and combining corrected SUV with CT respectively.There was a positive correlation between SUV,corrected SUV and the size of lesions(r 1 =0.536,P <0.001;r 2 =0.637,P <0.001).Conclusion Measur-ing corrected SUV appears to improve the accuracy of FDG PET/CT for characterization of peripheral solitary pulmonary lesions . The volume of lesion may potentially affect the value of SUV.

Objective To investigate the effect and mechanism of high concentration glucose on cholesterol absorption of human colon cancer epithelial Caco-2 cells.Methods The experimental study was used.(1) CCK-8 detected cell proliferation:the proliferation rate changes of Caco-2 cells were detected by CCK-8 when different concentrations (12.5,100.0,300.0,700.0,1 000.0,1 388.0 mmol/L) of glucose solution effects on Caco-2 cells in order to ensure the half hindering concentration of glucose concentration on Caco-2 cells.(2)Cholesterol absorption of Caco-2 cells was detected:Caco-2 cells were divided into the cholesterol group,cholesterol plus ezetimibe (cholesterol inhibitor) group and blank control group.Cholesterol group:100 μmol/L cholesterol solution and different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added.Cholesterol plus ezetimibe group:100 μmol/L ezetimibe,100 μmol/L cholesterol solution and different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added.Blank control group:DMEM culture medium and corresponding concentrations of DMSO were added.The cholesterol absorption amounts of Caco-2 cells were measured.(3) The relative expressions of ATP binding cassette G8 (ABCG8),ATP binding cassette G5 (ABCG5),Nickman-Pick CI Like 1 (NPC1L1) and scavenger receptor class B type Ⅰ (SR-B Ⅰ) were examined by Western blot in the different groups.Caco cells were divided into the glucose group,glucose plus ezetimibe group and control group.The different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added into the glucose group,different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution and 100 μmol/L ezetimibe were added into the glucose plus ezetimibe group,and 100 μmol/L ezetimibe were added into the control group.The relative expressions of ABCG8,ABCG5,NPC1L1 and SR-B Ⅰ were detected by Western blot.Measurement data were presented as (x) ±s,repeated measure variance analysis was used to perform variation trend test,and t test was utilized to conduct comparisons among groups.Results (1) CCK-8 results showed:proliferation rates of Caco-2 cells with the glucose solution concentration of 12.5,100.0,300.0,700.0,1 000.0 and 1 388.0 mmol/L were 1.380 ±0.043,1.238 ±0.072,0.736 ±0.035,0.336 ±0.021,0.316 ±0.020 and 0.288 ±0.010,respectively,with a statistically significant difference in the proliferation rates (F =11.019,P ＜ 0.05).The half hindering concentration of glucose solution on Caco-2 cells was 283.54 mmol/L.(2)Cholesterol absorption of Caco-2 cells:① the cholesterol absorption amounts of Caco-2 cells with the glucose solution concentration of 5.0,25.0 and 50.0 mmol/L were 0.282 ± 0.042,0.380 ± 0.063,0.390 ± 0.060 in the cholesterol group and 0.042 ± 0.012,0.197 ± 0.015,0.277 ± 0.029 in the cholesterol plus ezetimibe group,respectively,showing a statistically significant difference between the 2 groups (F =55.566,P ＜ 0.05).②There was a statistically significant difference in cholesterol absorption amounts of Caco-2 cells with different glucose solution concentration in the cholesterol group (F =79.117,P ＜ 0.05).The cholesterol absorption amounts of Caco-2 cells with the glucose solution concentrations of 5.0 mmol/L was lower than that with the glucose solution concentrations of 25.0 mmol/L and 50.0 mmol/L,respectively (t =11.207,11.532,P ＜0.05).There was no statistically significant difference in the cholesterol absorption amounts of Caco-2 cells between the glucose solution concentrations of 25.0 mmol/L and 50.0 mmol/L (t =12.389,P ＞ 0.05).③There were statistically significant differences in cholesterol absorption amounts of Caco-2 cells with the glucose concentration of 5.0 mmol/L and 25.0 mmol/L between cholesterol group and cholesterol plus ezetimibe group (t =10.908,10.644,P ＜ 0.05).(3) The results of Western blot showed:① the relative expression of NPC1L1 protein in Caco-2 cells with the glucose solution concentrations of 5.0,25.0 and 50.0 mmol/L were respectively 0.277 ±0.019,0.558 ±0.015,0.576 ±0.003 in the glucose group and 0.057 ±0.002,0.054 ±0.005,0.077 ±0.005 in the glucose plus ezetimibe group,showing a statistically significant difference (F =482.207,P ＜0.05).② The relative expression of NPC1L1 protein of Caco-2 cells with the different concentration of glucose solution in the glucose group were compared,with a statistically significant difference (F =8.112,P ＜ 0.05).There was a statistically significant difference in the relative expression of NPC1L1 protein in Caco-2 cells with the different concentration of glucose solution in the glucose plus ezetimibe group (F =11.708,P ＜ 0.05).③ The relative expression of NPC1L1 protein in Caco-2 cells with the glucose solution concentrations of 5.0,25.0 and 50.0 mmol/L in the glucose group was statistically different from that in the glucose plus ezetimibe group (t =8.112,11.708,13.920,P ＜ 0.05).Conclusion High concentration glucose solution could promote the reabsorption of cholesterol through increasing NPC1L1 protein expression in Caco-2 cells,and increase the risk of suffering from cholelithiasis in diabetes patients.

AIM:To observe the functional expression of calcium-sensing receptor (CaSR) in the mouse em-bryonic stem cells (mESCs).METHODS:The expression and distribution of CaSR were detected by Western blotting and immunofluorescence observation in 129 mouse ES-D3 cells.The intracellular concentration of free calcium ([Ca2+]i) was determined by confocal laser scanning microscopy .The cell viability was analyzed by MTT assay and flow cytometry .RE-SULTS:CaSR protein was expressed in mESCs .Extracellular calcium or neomycin significantly increased the expression of CaSR and [Ca2+]i.Neomycin increased the cell viability , up-regulated the protein expression of p-ERK2.These effects of neomycin were inhibited by NPS2390.CONCLUSION:CaSR is expressed in mESCs .The activation of CaSR is involved in the proliferation of mESCs .

OBJECTIVETo research the effect of polysorbate 80 (Tween 80) on Yuxingcao injection and volatile oils from Houttuynia cordata.

METHOD1H-NMR spectra of aldehydic and new matter in Yuxingcao injection, volatile oils of H. cordata, and solutions of Tween 80 and volatile oil of H. cordata are determined and compared from various angles of growing origin, storage temperature, and storage time.

RESULTThree aldehydic singlets in 1H-NMR spectra of every volatile oil from 4 aerial part of H. cordata were observed. These aldehydic peaks were basically disappeared and a new peak at delta 8.30 was found in 1H-NMR spectra of the volatile oil solutions in tween 80. Any obvious aldehydic peak in 1H-NMR spectra did not be observed in Yuxincao injection. A weak peak at 8 8.30 was found in 1H-NMR spectra in Yuxincao injection, and the peak high of delta 8.30 was remarked gone up when the injection was stored in 40 degrees C for 1 to 3 months.

CONCLUSIONTween 80 might cause the obvious reduce of aldehydic compounds contents and the production of a novel singal at delta 8.30 in 1H-NMR spectra when it was mixed with the volatile oil from the aerial part of H. cordata. The novel signal at delta 8.30 in 1H-NMR spectra existed in Yuxincao injection and was very small, but was increased remarkably when the Yuxincao injection was stored at 40 degrees C for 1 month at least.

Chemistry, Pharmaceutical ; instrumentation ; Drugs, Chinese Herbal ; chemistry ; Houttuynia ; chemistry ; Oils, Volatile ; chemistry ; Plant Oils ; chemistry ; Polysorbates ; Temperature

Objective To purify native F1 antigen from E pestis EV76 strain and determine its ef-ficacy against Y. pestis. Methods A new purification method was developed by the substitution of physical disruption ( glass beads) for organic solvent ( acetone and toluene) one, followed by a combination of ammo-nium sulfate fractionation and SephacrylS-200HR column filtration chromatography. Groups of mice were im-munized with F1 antigen adsorbed to 25% aluminum hydroxide in PBS by intramuscular route. The immu-nized animals were challenged subeutaneously(s, c. ) with 104 CFU of Y. pestis strain 141 at 18 weeks after the primary immunization. Results There was no IgG titre difference between two groups of mice with one-dose immunization, whereas in the two-dose immunization groups, the group F1-40 μg induced a statistically higher antibody titre than the group F1-20 μg. Complete protection was observed for animals immunized with purified F1 antigen by s.c. route. In contrast, the control mice immunized with aluminum hydroxide suc-cumbed to a same dose of Y. pestis 141 challenge. Conclusion This purification strategy is a simple and ef-fective, and can be operated in a large scale. Native F1 antigen extracted from Y. pestis EV76 is highly im-munagenic, and can be used as a key antigen component to develop sub-unit vaccine of plague.