A PCR assay used for detection and quantification of Neotyphodium lolii in tissues of the grass Lolium perenne L. was developed. Compared with the Microscopic analysis of stained tissues, this method was more accurate. For quantitative applications, real time PCR was used to quantify Neotyphodium lolii in Lolium perenne L.. The results showed that there were some variations between different individuals, but no variance existed between different tillers from the same plant. Thus it can be concluded that fungal content in the plant is not only associated with species/variety specific, but also with the genotype of the host plant as well.

Adult ; Female ; Hemofiltration ; Humans ; Hydrocarbons, Chlorinated ; poisoning ; Male ; Middle Aged ; Poisoning ; therapy ; Treatment Outcome

Objective To analyze if there are membrane estrogen receptors(ER)in endometrial carcinoma and if there is some relationship between membrane ER and nuclear ER.Methods The cell membrane and total cell ER?and ER?expressions of high and moderate differentiation endometrial carcinoma cells(Ishikawa and HEC-1A cells)were analyzed.Intermittent immunofluorescence dyeing and fluorescent microscopy were carried out with the ceils treated with polyformaldehyde and Triton X-100. Intermittent immunofluorescence dyeing and flow cytometry were carried out with the live cells and the cells treated with Triton X-100 respectively.Results There were fluorescences on the membrane of the Ishikawa and HEC-1A cells which were treated with polyformaldehyde.When the cells were treated with Triton X- 100,the fluorescences were also seen inside the cells.The fluorescence intensity of ER?and ER?in Ishikawa cell membrane(1.09?0.21,1.27?0.33)was stronger than the control,but there were no significant differences(P＞0.05).When treated with Triton X-100,the total cell fluorescence intensity of ER?and ER?in Ishikawa cell(4.21?0.34,4.69?1.96)was stronger than the membrane(P＜0.05). The ER?and ER?fluorescence intensity of HEC-1A cell membrane(1.58?0.13,1.49?0.04)were stronger than the control(P＜0.05).The fluorescence intensity of ER?and ER?of the HEC-1A cell (2.34?0.33,2.52?0.15)was stronger than the membrane also(P＜0.05).The membrane ER?fluorescence intensity of Ishikawa was lower than HEC-1A(P=0.028).But the total cell ER?fluorescence intensity of Ishikawa was higher than HEC-1A(P=0.002).Conclusions There are membrane ER on endometrial carcinoma cells Ishikawa and HEC-1A.The membrane ER must have some similarity to the nuclear receptor.There is no direct correlation between the quantity of the membrane ER and nuclear ER.

Objective To examine the effect of rapid infusion test guided by extravascular lung water index (EVLWI) on hemodynamics in critically ill patients at different states in order to guide volume resuscitation.Methods A prospective observation was conducted.Forty critically ill patients admitted to Department of Critical Care Medicine of Tianjin Third Central Hospital from June 2012 to April 2014 were enrolled.Based on the levels of EVLWI and pulmonary vascular permeability index (PVPI) and the cardiac function,the patients were divided into four groups:septic patients with normal EVLWI and PVPI (n =17),septic patients with increased EVLWI and PVPI (n =3),septic patients with increased EVLWI and normal PVPI (n =4),and coronary heart disease and heart failure patients with normal EVLWI and PVPI (n =16).The rapid infusion test was conducted in all patients using lactated Ringer solution 250 mL,followed by infusion of crystalloid with rate of 150 mL/h.The conditions of mechanical ventilation and vasoactive drugs were not changed during study.The changes in EVLWI,intrathoracic blood volume index (ITBVI),and cardiac index (CI) before capacity load,at immediate capacity load,and 15,45,105 minutes after load were determined by pulse indicator continuous cardiac output (PiCCO).On the base of volume status before and after the liquid infusion,the standard for the changes were:stroke volume (SV) increased by 12％-15％,central venous pressure (CVP) greater ≥ 2 mmHg (1 mmHg =0.133 kPa),CI ＞ 15％,and ITBVI change greater than 10％.Results There were no statistically significant differences in the observed indicators at the each time point before and after rapid infusion test among the four groups (all P ＞ 0.05).In septic patients with normal EVLWI and PVPI group,ITBVI was slightly increased by 5.4％-9.7％ from 15 minutes to 45 minutes after rapid infusion test.In coronary heart disease and heart failure patients with normal EVLWI and PVPI group,the EVLWI was increased by 11.9％,5.9％,and 14.7％ respectirely at 15,45,and 105 minutes,ITBVI was slightly increased by 6.4％ at 45 minutes,CI was increased by 29.5％ immediately after rapid infusion.In septic patients with increased EVLWI and PVPI group,CVP was increased by 8 mmHg immediately,EVLWI was increased significantly by 15.8％ at 45 minutes,ITBVI was slightly decreased by 10.0％ at 45 minutes,CI was increased by 24.7％ immediately,and increased by 17.0％ at 105 minutes,and PVPI was increased by 15.6％-28.1％ at 15-105 minutes after rapid infusion.In septic patients with increased EVLWI and normal PVPI group,CVP was increased by 1.5 mmHg at 15 minutes,EVLWI was increased immediately,which was increased by 17.4％,24.0％,and 31.4％ respectively at 15,45,and 105 minutes,ITBVI was increased by 13.9％ at 15 minutes,CI was increased by 16.1％ at 15 minutes after rapid fluid infusion.Conclusions Rapid fluid replacement in critically ill patients with crystalloid,regardless of whether the EVLWI was normal or increased,the short-term response was affected by the volume and cardiac function of patients.Different status of patients showed different volume effect curve:no significant changes in hemodynamic parameters were found in patients with normal EVLWI and volume parameters.In patients with potential cardiac dysfunction,CI and EVLWI increased significantly; regardless of PVPI increased or normal,EVLWI and CI were increased in patients with elevated EVLWI; two different changes could be found in the two types of pulmonary edema while ITBVI was increased.

In order to investigate the biological effects of the VP22?-mE6?/mE7 built-in adjuvant fusion protein vaccine on the tumor associated with HPV-16 infection.HSV-1 VP22? and HPV-16 mE6?/mE7 genes were cloned,and the pET28a-VP22?-mE6?/mE7 recombinat prokaryotic expression vector was constructed.Vector was transformed into Rosetta(DE3)E.coli string and expressed under the induction of IPTG.The re-naturalized protein was then purified via Ni2+ affinity adsorbent column and identified by SDS-PAGE and Western blot.Purified protein was immunized BalB/C and C57BL/6 mice to evaluate the immunogenicity and anti-tumor activity.The expressed recombinant protein formed as inclusion body with a prediction MW about 34kDa and contained approximately 45% of total somatic protein.The VP22?-mE6?/mE7 immunization induced higher titer of specific IgG against HPV,higher level of lymphocyte proliferation and better effect on suppressing HPV16 positive TC-1 tumor growth than the mice immunized with mE6?/mE7 alone.The results showed that the recombined built-in adjuvant vaccine could induce specific cellular immune response in vitro and inhibit the TC-1 tumor proliferation in vivo,that would be a foundation for further studies and developments of inner adjuvant vaccines.

Objective To investigate the effect of bundle of care in the prevention of lower extremities deep venous thrombosis ( DVT) among patients with severe ischemic stroke .Methods Two hundred and twenty-eight patients with severe ischemic stroke in the Neurological Intensive Care Unit (NICU) in 2012 were selected as experimental group , and 206 patients with severe acute ischemic stroke in NICU in 2011 as control group.In the experimental group, the patients were treated with bundle of care , which included preventive screening, body position nursing care, nutrition support and liquid management , vascular protection, physical prevention, medicine prevention, passive movement and other measures .And in control group, conventional nursing was applied.The incidence of DVT was observed and compared in two groups .Results There were only 8 patients (3.51%) with lower extremities DVT in the experimental group , and 49 patients (23.79%) with lower extremities DVT in the control group , the difference was statistically significant between the two groups (χ2 =39.004 1,P <0.01).However, there was no statistically significant difference in the incidence of unilateral and bilateral limbs paralysis (χ2 =0.152 9,P=0.696).Conclusions The application of bundle of care can effectively reduce the incidence of deep venous thrombus of lower extremity in patients with severe ischemia stroke.It is worth for clinical use.

Abstract: Objective To understand the mental health literacy level of residents in Hainan Province, and to provide evidence for promoting mental health promotion and improving the mental health literacy level of residents. Methods Multi-stage random sampling method was used to investigate The National Mental Health Literacy Questionnaire among 6 895 residents in 12 districts including Changjiang, Chengmai, Dongfang, Ledong, Lingshui, Haikou Longhua District, Haikou Meilan District, Qionghai, Sanya, Tunchang, Wenchang and Wuzhishan. Results　In the survey 6 895 residents in 12 counties, cities, districts of Hainan Province, 365 of them reached the standard of mental health with the standard rate of 5.3%. The mental health knowledge score was (54.00±17.02) with the standard rate of 7.7%; the self-assessment score was (26.11±3.99) with the standard rate of 75.9%; the score of mental health skill was (26.22±7.25) with the standard rate of 44.8%. The mental health literacy level of medical workers was the highest, with the standard rate of 13.5% (112/830), and the mental health literacy level of farmers was the lowest, with the standard rate of 0.8% (13/1 647). The results of unconditional Logistic regression showed that the factors entering the regression model included educational background (OR=2.268), personal monthly income (OR=1.129), gender (OR=1.302), household registration (OR=0.776), and whether they had participated in mental health related courses OR training (OR=0.511). The higher the educational background and personal monthly income, the higher the psychological quality. The mental health of women was higher than that of men, and that of urban was higher than that of rural, and those who had participated in mental health related courses were higher than those who had not. Conclusions The mental health literacy level of Hainan residents is at a low level, and the influencing factors are multifaceted. It is suggested to strengthen the mental health monitoring comprehensively and carry out rich health education service mode for different groups.

Animals ; Blotting, Western ; Cell Differentiation ; drug effects ; Embryonic Stem Cells ; cytology ; enzymology ; Isoenzymes ; analysis ; Mice ; Neurons ; enzymology ; Protein Kinase C ; analysis ; Tretinoin ; pharmacology

OBJECTIVETo investigate the effect of Suanzao nacute hepatic failure in mice.

METHODAcute liver failure was induced in male Kunming strain mice by enterocoelia injecting the animals with D-Gal-N and LPS. The mice in treatment groups were given corresponding drug 2 h before administration of D-Ga1-N and LPS, and the mice in control group were given the same dose of distilled water. The 24 h survival rate, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) levels were compared. Serum the levels of TNF-alpha and IL-1 and the levels of SOD, MDA, GR, GSH, NO and NOS in the liver were determined.

RESULTTreatment with suanzaoren decoction could increase the survival rate and improve the liver histological feather. Suanzaoren decoction inhibited the serum the levels of ALT, AST, TNF-alpha and IL-1, and reduced the levels of MDA, NO and NOS and increased the levels of GR and SOD in the liver.

CONCLUSIONTreatment with Suanzaoren decoction can suppress the D-Gal-N/LPS-induced acute hepatic failure. It may be the mechanism that Suanzaoren decocotion regulate the production of inflammatory cytokines and free radicals.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Cytokines ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Free Radicals ; metabolism ; Galactosamine ; Glutathione ; metabolism ; Lipopolysaccharides ; Liver ; drug effects ; metabolism ; pathology ; Liver Failure, Acute ; blood ; chemically induced ; prevention & control ; Male ; Malondialdehyde ; metabolism ; Mice ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Random Allocation ; Superoxide Dismutase ; metabolism

OBJECTIVEThis study examined the effects of PI3K inhibitor LY294002 on the differentiation of mouse preadipocytes and the expression of CCAAT enhancer binding protein α (C/EBPα) and peroxisome proliferation activated receptor γ (PPARγ), in order to study the possible roles of insulin receptor substrate (IRSs)/PI3K signal pathway in the differentiation of preadipocytes.

METHODSThe mouse 3T3-L1 cells were cultured normally and divided into experimental and control groups. 3T3-L1 cells in the experimental group were treated with PI3K inhibitor LY294002 (25 μmol/L) and those in the control group were treated with DMSO culture medium. 3-isobutyl-1-methylxanthine (IBMX) (0.5 mmol/L), dexamethasone (10-6 mol/L) and insulin (5 μg/mL) were used to induce the differentiation of 3T3-L1 preadipocytes in both groups. Before culture, and 2, 4 and 8 days after culture, the cells were collected to detect the expression of C/EBPα and PPARγ by real-time PCR and Western blot assays. The lipid droplets of 3T3-L1 preadipocytes were observed by oil-red O staining.

RESULTSPI3K inhibitor LY294002 did not affect the expression of C/EBPα and PPARγ in un-induced 3T3-L1 preadipocytes (P>0.05), but decreased the expression of C/EBPα and PPARγ during the in vitro induced differentiation of 3T3-L1 preadipocytes compared with the control group (P<0.05 or 0.01). The lipid droplets count was greatly reduced by LY294002.

CONCLUSIONSPI3K inhibitor LY294002 can inhibit the differentiation of mouse 3T3-LI preadipocytes and the expression of C/EBPα and PPARγ in the differentiation of 3T3-LI preadipoeytes, suggesting that IRSs/PI3K signal pathway may play an important role in the differentiation of 3T3-L1 preadipocytes by regulating the expression of C/EBPα and PPARγ.

3T3-L1 Cells ; Adipocytes ; cytology ; drug effects ; Animals ; CCAAT-Enhancer-Binding Protein-alpha ; analysis ; genetics ; Cell Differentiation ; drug effects ; Chromones ; pharmacology ; Gene Expression Regulation ; drug effects ; Mice ; Morpholines ; pharmacology ; PPAR gamma ; analysis ; genetics ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; RNA, Messenger ; analysis