Objective: To study the chemical constituents of aerial part of Rosmarinus officinalis. Methods: Compounds were isolated and purified by repeated column chromatography on silica gel column and Sephadex LH-20 gel column. The structures of the new compounds were elucidated by analysis of their NMR and HRESIMS data. The cytotoxicity of the compounds was evaluated by SRB methods. Results: Ten compounds were obtained and identified as 7-O-isopropyl-epirosmanol (1), 7-O-isopropyl rosmaquinone (2), rosmanol (3), 7-O-methyl-rosmanol (4), epirosmanol (5), 7-methoxy-epirosmanol (6), rosmaquinone B (7), carnosol (8), galdosol (9) and pisiferal (10). Conclusion: Compounds 1 and 2 are new abietane diterpenes named 7-O-isopropyl-epirosmanol (1), 7-O-isopropyl rosmaquinone (2), and showed no cytotoxicity.

Objective To quantitatively measure the perfusion parameters of placenta in different stage of pregnant rats using contrast-enhanced ultrasonography (CEUS) with contrast pulsed sequencing (CPS), and to analyze the relationship between perfusion parameters and changes of placental vascular bed. Methods Sixty healthy pregnant rats in according to the requirements of the experiment was divided into three groups: 15 days, 17 days and 20 days of gestation with 20 animals in each group. One blous injection of SonoVue (Sonovue 1.0 ml/kg) via a tail vein was administered to each rat, and the time-intensity curves (TIC) of placenta and uterine muscle wall were drawn with ACQ using CPS technique with MI 0.20, and the perfusion parameters were calculated. Then 4 μm vertical placenta sections were cut and stained with hematoxylin and eosin. Surface area densities of placental maternal blood space was measured with image analysis software. Results The peak intensity (PI) of 17 days and 20 days was higher than that of 15 days pregnant rats (P＜0.05). There was no difference in PI between 17 days and 20 days (P＞0.05), and nor of arrivel time (AT) and time-to-peak (TTP) (P＞0.05) among the three groups. There was significant difference of surface area densities of placental maternal blood space among the three groups (P＜0.05). PI was positively correlated to the surface area densites of placenta (P＜0.05). Conclusion There is close relationship between peak intensity and area densities of placental maternal blood space. CPS technique can sensitivly detect changes of the placenta vascular bed in different stage of pregnant rats.

Objective To evaluate the perfusion parameters in different zone of placenta using CPS,to research placental blood perfusion of different zone. Methods A total of 60 pregnant rats were divided into 3groups, 15 day,17 day and 20 day of gestation with 20 animals in each group,the placenta was divided into the central zone and marginal zone. SonoVue was injected by tail vein of rat using CPS, the time-intensity curves of the central zone and marginal zone were drawn and perfusion paramcters were calculate by software ACQ and Sonoliver. Surface area densities of different zone of placental maternal blood space was measured with histoligical method,and using immunohistochemical marked the placenta vascular. Results Arrivel time(AT) and time-to-peak(TTP) of placenta central zone was earlier than marginal zone,the peak intensity(PI) of the central zone was higher than that of the marginal zone ( P ＜ 0.05). There was significant difference between the central zone and marginal zone of surface area densities of placental maternal blood space ( P ＜0.05). PI of the central zone and marginal zone and the surface area densites of placenta was positively correlated ( P ＜0.05). The placental blood vessels was not expressed by factor Ⅷ.Laminin in the placental basement membrane expression of 20 day of gestation was not continuous or missing. Conclusions The surface area densities of central zone and marginal zone was different. CPS technique can be sensitive to detect changes of the placenta vascular bed in different zone.

A sensitive and reliable liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS) method was established to simultaneously quantitate four categories of compounds (isoflavonoids, flavonoids, alkaloids and saponins) in Gegen-Qinlian decoction (GQD). These compounds were separated by a Shiseido CAPCELL PAK C18 column with a linear gradient consisting of 0.1% (v/v) formic acid in water (A) and 0.1% (v/v) formic acid in acetonitrile (B), and delivered at a flow rate of 0.3 mL/min. All the analytes were determined by electrospray positive ionization tandem mass spectrometry in a multiple reaction monitoring (MRM) mode. Linearity, accuracy, precision, recovery and stability of the method were evaluated with the validation over the range of 4.0-538 5 ng/mL. The proposed method was applied to the analysis of a Chinese herbal preparation GQD successfully.

The prevalence of inflammatory bowel disease (IBD) in China is increasing year by year, however, the efficacy and safety of commonly used therapeutic methods are limited.Granulocyte and monocyte adsorptive apheresis (GMA) is one of the effective methods for treatment of IBD used abroad, however, there is still lacking of such research in China.Aims: To investigate the efficacy and safety of GMA in IBD patients.Methods: A retrospective study was conducted in 21 cases of IBD patients [13 cases with ulcerative colitis (UC) and 8 with Crohn's disease (CD)] who accepted GMA treatment from May 2013 to July 2014 at the Shanghai Rui Jin Hospital.All the cases were poor responders to 5-aminosalycylic acid (5-ASA) or steroid-refractory.The clinical data were collected, and the clinical activity index (CAI), endoscopic activity index (EAI), laboratory parameters including serum albumin (Alb), hemoglobin (Hb), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), leukocyte count and percentage of neutrophils, as well as the adverse effects before and two weeks after the end of GMA treatment were analyzed.Results: After GMA treatment, both CAI and EAI were decreased significantly in UC and CD groups as compared with those before treatment (P all <0.05).Among laboratory parameters, Alb was increased in UC group and CRP was decreased in both UC and CD groups after treatment (P all <0.05).No significant differences were found in other laboratory parameters in both UC and CD groups before and after treatment (P all >0.05).The treatment was well tolerated with no severe adverse effects.Conclusions: GMA is safe and effective for ameliorating clinical symptoms, attenuating intestinal mucosal injury and controlling active inflammation in IBD patient that has not responded to 5-ASA or steroid treatment.Prospective clinical studies with large samples are needed to confirm these findings.

BACKGROUND:Previous studies have found that estrogen deficiency causes a reduction in the activity of bone marrow mesenchymal stem cel s (BMSCs). OBJECTIVE:To explore the effect of endothelial progenitor cel s (EPCs) on the BMSCs proliferation and apoptosis ability of osteoporosis rats. METHODS:Healthy female Sprague-Dawley rats, 6 weeks old, were enrol ed and subjected to bilateral ovariectomy to make osteoporosis models. BMSCs and EPCs were isolated using density gradient centrifugation combined with adhesion method, and identified with surface markers, cel proliferation and immunocytochemistry in vitro. We used Transwel inserts to establish EPCs and OVX-BMSCs indirect co-culture system. Control groups were OVX-BMSCs group and sham-BMSCs group in which rats were only subjected to remove the equal amount of fat tissues around the ovary. Flow cytometry was applied to detect BMSCs proliferation and apoptosis ability. RESULTS AND CONCLUSION:Compared with the control groups, the results of flow cytometry test showed that the proportion of OVX-BMSCs at S phase was significantly increased at 3 days of indirect co-culture with EPCs and the apoptosis rate was significanty reduced at 10 days of indirect co-culture with EPCs (both P<0.05). These results suggest that EPCs can promote the proliferation but inhibit the apoptosis of OVX-BMSCs.

Background Researches showed that wild-type p53(wtp53)and Rb94 genes inhibit the growth of retinoblastoma(RB),and these genes are involved in signal pathway in the induction and maintenance of cellular senescence.Thus the combination of two genes to inhibit growth of RB is concerned.Objective This study was to observe the inhibitory effect of the co-transfection of Rb94 and wtp53 gcnc into subretina on RB with ultrasound microbubble.Methods HXO-Rb44 suspension was subretinally injected to establish the RB model in 40 SPF female Balb/c nude mice.The RB models were randomized into model control group,wtp53 group,Rb94 group and wtp53+Rb94 group,and 0.1 ml relevant gene microvesicle suspension was injected via caudal vein in the different groups,but no any gene was used in the model control group.Seven days after modeling,followed by 0.5 W/cm2ultrasonic wave irradiated the eyeballs immediately for 4 seconds ×2 times and interrupted for 24 seconds.Eyeballs were extracted 7 days after gene transfection,and the expressions of wtp53 mRNA and Rb94 mRNA in tumor tissuc were detected by RT-PCR,and wtp53 and Rb94 protein in tumor tissue were assayed using Western blot.Immunochemistry was used to exam the VEGF expression,and TUNEL was used to evaluate the apoptosis of the tumor cells.Results The model successful rate after HXO-Rb44 suspension was 80％ (32/40)and obvious malformation cells were seen under the light microscope.In 7 days after gene transfection,no response band for wtp53 mRNA and Rb94 mRNA were found.The relative expression valuc of wtp53 mRNA was 0.65±0.07 in the wtp53 group,and that in wtp53+Rb94 group was 0.32±0.02,showing a significant difference between them (t =11.743,P =0.000).Rb94mRNA relative value was 0.42 ±0.03 in Rb94 group,and that in the wtp53 + Rb94 group was 0.23 ± 0.03,with a significant difference(t=5.041,P=0.001).The response bands of wtp53 and Rb94 proteins were seen in wtp53group,Rb94 group and wtp53+Rb94 group.Immunochemistry showed that the positive reactive intensity for VEGF in tumor tissue was obviously weaker in wtp53+Rb94 group than that in the wtp53 group,Rb94 group and model control group.Apoptotic index(Al) was 37.35±2.14 in the wtp53+Rb94 group,showing a significant increase in comparison with model control group (0.46 ± 0.05),wtp53 group (5.05 ± 0.80) and Rb94 group (6.43 ± 1.02) (t =-34.395,-28.206,-26.006,P＜0.01).Conclusions Ultrasound microvesicle enable double gene transfecting into RB tumor tissue,and Rb94 gene cooperation with wtp53 gene enhance the inhibitory effect on RB by promoting the apoptosis of RB cells.

Objectives： The current study was designed to identify mimic polypeptide epitopes of lung cancer-related antigen WLA-Ag1 through screening random peptide library which was on display in phages, and provide useful information for further study of the characterization of WLA-Ag1 antigen and the potential chance for developing new diagnostic or therapeutic methods for lung cancer. Methods： Through affinity enrichment and immunoscreening of phage-displayed fifteen-peptide libraries with a monoclonal antibody named WLA2C4 against WLA-Ag1, the enriched phages was obtained and plated on agar plates. Some positive clones picked at random were confirmed by ELISA using WLA2C4 monoclonal antibody and DNA sequencing. The homologous comparison of amino acid sequences of positive clones was conducted through searching the protein sequence databank on Internet. The digestion analyses of amino acid sequences were also conducted by a software. Results： Twelve positive clones have been found with a consensual DNA sequence, that is, 5′ -GCT GGT TGG ATT ACT TTT CAT CGT CGT CAT CAT GAT CGT GTT CTT-3′ . Amino acid sequence was AGWITPHRRHHDRVL, with three trypsinase-digestible sites. This coincided with our previous finding that WLA-Ag1 can be digested by trypsinase. Conclusions： The amino acid sequence found in this study might be the epitope of WLA-Ag1. Screening of bioactive peptides from random peptide libraries using monoclonal antibodies as the ligands is an effective method to identify the epitopes of cancer-related antigens.

85 mL/min),CL=6.0?(WT/60)~(0.52).④The increased volume of peripheral distribution (V_2) was observed when norvancomycin was co-administered with diuretics;④Reduced drug clearance,prolonged t_(1/2),and increased values of AUC_(24) were found in elderly patients.Conclusions Renal function impairment and age have significant impact on PK parameters of norvancomycin.Dosing regimens of norvancomycin were finally established for different patients on the basis of important PPK parameters generated from different groups of patients.

Objective To evaluate the role of mitochondrial fusion?fission in endotoxin?induced a?cute lung injury in rats. Methods Twenty healthy male Sprague?Dawley rats, weighing 160-180 g, were e?qually and randomly divided into either control group ( group C ) or endotoxin?induced acute lung injury group (group L) using a random number table. Lipopolysaccharide 5 mg∕kg was injected intravenously in group L, while the equal volume of normal saline 0?5 ml was given instead in group C. The animals were sacrificed at 6 h after administration of lipopolysaccharide or normal saline. The lungs were immediately re?moved for measurement of wet to dry lung weight ratio ( W∕D ratio) , superoxide dismutase ( SOD) activity and malondialdehyde ( MDA) content. The mitochondrial fusion proteins mitofusin 1 ( Mfn1) , Mfn2 and op?tic atrophy 1 ( OPA1) mRNA and protein expression was detected, and mitochondrial fission proteins dy?namin?related protein 1 (Drp1) and fission 1 (Fis1) mRNA and protein expression was also detected in lung tissues. Results Compared to group C, the W∕D ratio and MDA contents in lung tissues were signifi?cantly increased, SOD activity was decreased, Mfn1, Mfn2 and OPA1 mRNA and protein expression in lung tissues was down?regulated, and Drp1 and Fis1 mRNA and protein expression was up?regulated in group L. The pathological damage to lung tissues was obviously aggravated in group L when compared to group C. Conclusion The mechanism underlying endotoxin?induced acute lung injury is related to enhanced oxidative stress responses caused by decreased mitochondrial fusion and increased mitochondrial fission in rats.