AIM To analyze the dynamic changes of five constituents in Ligustri lucidi Fructus at five picking time (August,September,October,November,December).METHODS The HPLC analysis of Ligustri lucidi Fructus ethanol extract was performed on a 25 ℃ thermostatic Aglient Zorbax SB-C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 1 mL/min in a gradient elution manner,and the detection wavelength was set at 224 nm.RESULTS Salidroside,tyrosol,luteolin-7-O-glucoside,ligustroflavone and specnuezhenide showed good linear relationships within their own ranges (r ＞0.999 0),whose average recoveries were 99.56％-100.30％ with the RSDs of 0.89％-1.23％.The contents of various constituents (except for tyrosol) were the highest in samples picked up in September,followed by those picked up in October.CONCLUSION The suitable picking time of Ligustri lucidi Fructus is September and October.

Background:Inflammatory bowel disease( IBD)is a group of chronic and non-specific intestinal inflammatory diseases of undetermined origin. Functional impairment of macrophages has been associated with the dysregulation of mucosal immunity in intestinal tract of patients with IBD. Aims:To investigate the correlation of serum levels of macrophage polarization-related cytokines with the development and disease activity of IBD. Methods:A total of 105 IBD patients admitted from May 2013 to May 2014 at Shanghai Ruijin Hospital were recruited,of them 65 were Crohn’s disease (CD)and 40 were ulcerative colitis( UC). Twenty-four patients with colonic polyps were served as controls. Serum samples were obtained and the levels of interleukin-1beta(IL-1β),IL-6,IL-10,IL-13,interferon-gamma(IFN-γ)and inducible nitric oxide synthase(iNOS)were determined by ELISA method. Results:Serum levels of IL-1β,IL-6,IL-10, IL-13 and IFN-γ were significantly higher in CD group than in control group(P < 0. 05),and serum levels of IL-10,IL-13 and IFN-γ were significantly higher in UC group than in control group(P < 0. 05). Logistic regression analysis revealed that serum IL-13(OR = 1. 009,P = 0. 005)and iNOS( OR = 0. 982,P = 0. 013)were correlated with CD,while Spearman correlation coefficient demonstrated a link between serum IL-10 and disease activity of CD(rs = - 0. 432,P =0. 014). No correlations were observed between serum levels of these cytokines and development and disease activity of UC (P > 0. 05). Conclusions:Serum levels of macrophage polarization-related cytokines increase to varying degrees in IBD patients,but these cytokines have no obvious correlations with IBD and its disease activity. Presumably,theses cytokines are only involved in but not the triggers in the development of IBD.

BACKGROUND:Segmental bone defects resulting from osteoporotic fractures, trauma, congenital bone dysplasia and progressive bone disorder are very common, and bone tissue engineering provides a new approach to bone defect repair. Growth factors related to bone tissue engineering bone have been reported a lot and have achieved some results. How to mimick the natural timing of different growth factors with different bioactivities has become the current hotspot in bone repair. OBJECTIVE: To review the new developments in vascular endothelial growth factor and bone morphogenetic protein in bone tissue engineering. METHODS: The first author searched CNKI (1990/2015) and Medline database (1990/2015) for related articles using the key words of “osteogenic factors, angiogenic factors, tissue engineering bone, bone repair, vascularization, vascular endothelial growth factor, bone morphogenetic protein, sequential release, seed cels, cytoskeleton” in Chinese and English, respectively. Mechanism of action and research direction about vascular endothelial growth factor and bone morphogenetic protein were summarized. RESULTS AND CONCLUSION:Totaly 313 papers were searched initialy, and finaly 87 papers were enroled in result analysis. The results show that different growth factors play different roles in bone repair. Vascularization and osteogenesis are the most important processes in bone repair. The osteogenic factors play an important role in maintaining bone structure and bone formation. The angiogenic factors can provide oxygen and nutrients for tissue growth, differentiation and functionalization. The combination of osteogenic and angiogenic factors has a better osteogenic effect than osteogenic or angiogenic factors used alone. However, the most important problem is how to control the exogenous osteogenesis and the release dosage of angiogenic factors in bone repair.

Objective To analyze the clinical characteristics,pathological classification and therapy strategy of rectal carcinoid and its prognostic factors.Methods Forty four patients with rectal carcinoid were diagnosed and treated in Ruijin Hospital Shanghai Jiaotong University from January 2006 to November 2013,among whom 21 patients (19 males and 2 females) were followed-up for 1-7.5 years.The clinical data of these cases were retrospectively analyzed.Results The patients underwent colonoscopy because of changed bowel habits and/or abnormal digital rectal examination,none of them had carcinoid syndrome.Colonoscopy showed that most lesions presented yellowish in color and smooth in surface; the diameter of the tumor was ≤ 1.0 cm in 12 cases (57％) ; the tumors were located at the rectum within 8 cm from anal rim in 17cases (76％); most of them were well differentiated.Immunohistochemistry demonstrated that NSE expression was highly positive.The 1 year-and 3year-survival rate were both 100％.Among 8 cases who were followed up for over 5 year,2 relapsed.Conclusions The digital rectal examination plays a key role in detecting rectal carcinoid.Though prognosis is relatively good,we should keep close following-up to detect the recurrence.The main risk factors influencing the prognosis are tumor size,depth of invasion and clinical stage.The combination measurement of CEA,TSGF,CA19-9 and NSE will either increase the sensitivity or the specificity of early detection.

AlM:To explore the primary culture conditions for four kinds of lens epithelial cells ( LECs) of rat, rabbit, dog, and human, and measure their growth characteristics.METHODS:The lens capsule or anterior capsular tissue of rat, rabbit, dog and patient were removed by different methods, and they were cut into tiny pieces for primary culture by modified tissue adherent method. The morphological features of four kinds of LECs were observed under an inverted microscope.RESULTS: Four kinds of LECs of rat, rabbit, dog and human could be cultured primarily by tissue adherent method. With the evolution of tissue source, the adherent capacity of LECs gradually strengthened, cells form were changed from irregular polygon to oval, nucleus rounded and cytoplasm enriched gradually. Four kinds of LECs had fibrotic changes after several passages.CONCLUSlON: LECs of rat, rabbit, dog and human can be primarily cultured. This method lays the foundation for the mechanism research of caratact and related fields on the cellular and molecular levels.

Objective To construct a new conditionally replicative adenovirus (CRAds) targeting carcinoembryonic antigen (CEA) positive colorectal cancer cells. Methods The DNA fragment of the CEA gene promoter was amplified through PCR and cloned into the vector carrying fusion reporter gene EGFP-Luc to construct expression plasmid pCEA-EGFPLuc. The constructed plasmid pCEA-EGFPLuc was transfected into CEA positive and negative cells by liposome. The activity of CEA gene promoter was evaluated by detecting the expression of EGFP and luciferase activity. The conditionally replicative adenovirus Ad.CEA-E1A/CMV-TK carrying suicide gene HSVtk was constructed, in which the E1A gene was controlled under CEA promoter. CEA positive(Lovo and SW620)and negative tumor cells(HeLa) were infected with Ad.CEA-E1A/ CMV-TK. The selective cytotoxicity of Ad.CEA-E1A/CMV-TK and the synergistic effect of the virus with GCV in CEA positive tumor cells were evaluated by the expression of E1A, cytopathic effect and cell survival rate. Results CEA promoter possesses a good specificity as well as high activity. The expression of E1A only presented in CEA positive tumor cells. After infection with Ad. CEA-E1A/CMV-TK, the cell survival rates of Lovo and SW620 were (36.72±2.49)% and (39.82±4.76)%, respectively, significantly lower than that of Hela[(87.44±2.76)%1 (P<0.01). When combined with GCV, Ad.CEA-E1A/CMV-TK had better oncolytic effect on Lovo and SW620 cells, with cell survival rates of (17.26±3.65) % and (23.93±5.40) %, respectively, significantly lower than those without GCV[(36.72±2.49) % and (39.82±4.76) %, respectively] (P<0.01). Conclusion Ad. CEA-E1A/CMV-TK under the control of CEA promoter has selective cytotoxic effect on CEA positive colorectal cancer cells, and the effect can be enhanced when combined with GCV.

Background Adeno-associated virus-based vector is one of most efficient vehicles.It presents with a long-term and efficient transfer and expression of therapeutic genes with minimal toxicity.But its delayed-and low-efficient transgene expression limits the application of AAV vector.To explore an improving method of AAV infecting RPE cells is the hot spot. Objeetive Present study was to investigate whether adeno-associated virus (AAV)combined with low dose non-replieable adenovirus(Ad-null)can enhance its infection efficieney on RPE ceils in vitro. Methods Human RPE cells were isolated from the donate eyeballs under the approval of the Ethic Committee of this hospital.The cells were cultured in DMEM containing 10%fetal bovine serum.AAV particles with enhanced green fluorescence protein(EGFP)were added into the medium alone or in combination with different amount of adenovirus for 30 days.The cells were detected under the fluorescence microscope,and the protein expression levels of report gene EGFP in RPE cells were analyzed with Western blotting assay. Results Melanin granules could be found in cultured RPE cells.EGFP was expressed in RPE cells at 2 days after AAV-EGFP infection and peaked at 12 days and remained for about 3-week duration,showing the green influorescence under the influorescence mwroscope.After the cells were infected by AAV2-EGFP with 0.01 to 1000 MOI Ad-null respectively,the number of cells with green influorescence was obviously increased with the enhanced infiuorescence intensity.The enhance of the infection efficiency began in the 0.1 MOI Ad-null group and peaked in 10 MOI Ad-null group.Dead cells were exhibited in the 100 or more MOI Ad-nulor group.Western blotting assay demonstrated that the protein expression level of EGFP in RPE cells enhanced significantly in 1 and 10 MOI Ad-null groups compared with only AAV infection group. Conclusion These finding suggested that the infection efficiency of AAV can be improved significantly when it is used with low dose Ad-null in vitro.This offers a basis for further study of gene therapy.

OBJECTIVETo investigate the awareness and use of banned pesticides among vegetable greenhouse farmers in a town of Ledu County, Qinghai Province, China and to provide a basis for the management and control of banned pesticides.

METHODSLocal residents who lived in a town of Ledu County for more than 5 years were selected as subjects in August 2012. A questionnaire survey was conducted in 267 vegetable greenhouse farmers who were selected from five villages of the town by cluster random sampling.

RESULTSOf the 267 subjects, 249 (93.26%) completed and returned the questionnaires. Eighty-seven (34.9%) of the 249 subjects were aware of banned pesticides, and the banned pesticide awareness rate varied significantly among the subjects with different education levels (χ(2) = 11.061, P = 0.011). Only education level entered the regression equation in non-conditional logistic regression analysis. Only five (2.0%) of the 249 subjects knew the banned pesticides as well as the details. All the 249 subjects used banned pesticides to varying degrees in the past 5 years. Only 9.0% (17/189) of the vegetable greenhouse farmers knew the time pesticides were initially banned in China.

CONCLUSIONThe banned pesticide awareness rate is relatively low in the town of Ledu County, and this rate is influenced by education level. It is needed to spread the knowledge and hazards of banned pesticides.

Agriculture ; China ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Occupational Exposure ; prevention & control ; Pesticides ; Surveys and Questionnaires

OBJECTIVETo investigate the early prognostic values of arterial lactate and base excess (BE) in patients with paraquat poisoning.

METHODSSeventy-five patients with paraquat poisoning were divided into sudden death group (n = 10) who died within 24 h after admission, recent death group (n = 31) who died more than 24 h after admission, and survival group (n = 34). Arterial lactate and BE were measured on admission and at 24 h after admission. The prognostic values of arterial lactate and BE were analyzed.

RESULTSThe arterial lactate measured on admission was significantly higher in the sudden death group than in the recent death group and survival group (P < 0.01), but there was no significant difference in arterial lactate between the recent death group and survival group (P = 0.309). The BE measured on admission was significantly lower in the sudden death group than in the recent death group and survival group, and it was significantly lower in the recent death group than in the survival group (P < 0.01 or P < 0.05). At 24 h after admission, the recent death group had a significantly higher arterial lactate (P < 0.01) and a significantly lower BE (P < 0.01), as compared with the survival group. The logistic regression analysis showed that the two indices were significantly associated with prognosis (P < 0.01). On admission, the areas under the receiver operating characteristic (ROC) curve (AUCs) of arterial lactate and BE for predicting death were 0.692 and 0.787, respectively, and the cut-off values were 3.25 mmol/L and -1.75 mmol/L, respectively; the AUCs of arterial lactate and BE for predicting sudden death were 0.995 and 1, respectively, and the cut-off values were 7.1 mmol/L and -12.8 mmol/L, respectively. At 24 h after admission, the AUCs of arterial lactate and BE for predicting death were 0.743 and 0.822, respectively, and the cut-off values were 2.15 mmol/L and -5.55 mmol/L, respectively.

CONCLUSIONArterial lactate and BE have certain values in predicting the death, especially the sudden death, in patients with acute paraquat poisoning.

Adult ; Aged ; Arteries ; chemistry ; Female ; Humans ; Lactic Acid ; blood ; Male ; Middle Aged ; Paraquat ; poisoning ; Poisoning ; diagnosis ; Prognosis

OBJECTIVETo explore the mechanism of reactive oxygen species (ROS) in manganese chloride (MnCl(2))-induced apoptosis in PC12 cells.

METHODSThe model that MnCl(2) induced apoptosis in PC12 cells was established. The apoptotic effect of MnCl(2) on PC12 cells was analyzed with the MTT, the flow cytometry and the DNA fragmentation. The production of ROS and ATP in MnCl(2)-induced apoptosis of PC12 cells was examined. The influence of MnCl(2) on the expression of bcl-xl, bax and the activity of Caspase 3 was also analyzed.

RESULTSMnCl(2) triggered PC12 cells apoptosis in a dose-and time-dependent manner (P < 0.01). The rate of apoptosis was significantly increased (P < 0.01) when MnCl(2) of 2 mmol/L induced PC12 cells for 36 hours. The production of ROS was increased (P < 0.001) and the quantity of ATP was decreased (P < 0.01) in PC12 cells with the same inducement of MnCl(2). The expression of bcl-xl was inhibited and the bax was activated in this process (P < 0.01). Caspase 3 was also activated (P < 0.01).

CONCLUSIONMnCl(2) induces apoptosis of PC12 cells, which is related to the increase of ROS, the inhibition of the mitochondria and the activation of Caspase 3.

Adenosine Triphosphate ; biosynthesis ; Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Chlorides ; toxicity ; DNA Fragmentation ; drug effects ; Manganese Compounds ; PC12 Cells ; Rats ; Reactive Oxygen Species ; metabolism ; bcl-2-Associated X Protein ; biosynthesis ; bcl-X Protein ; biosynthesis