BACKGROUND: Apelin/APJ system has a wide range of physiological functions,but its intracellular signal transduction,in particular,apelin receptor desensitization,internalization,resensitization degradation,have still no consistent opinion.OBJECTIVE: To construct eukaryotic expression vector expressing human apelin receptor(APJ)tagged to red fluorescent protein(pDsRED-express-C1),and to determine the expression in human embryo kidney 293 cells.METHODS: The plasmid pcDNA3.1-hAPJ was used as a template for PCR amplification of human APJ.Following PCR amplification the PCR product were removed and enzymatic digestion with EcoR I and BamH I.Same enzymes were used to cut vector pDsRED-express-C1.The digestive product was ligated by conventional methods of connection,then transfected into Competent E.coli TOP10.Single clones were picked plasmid extraction,followed by restriction enzyme digestion and finally DNA sequencing.The recombinant plasmid with correct sequencing was transfected into human embryonic kidney cells,PI staining,followed by the observation under a confocal microscope.RESULTS AND CONCLUSION: PCR amplified a 1.2-kb fragment,which was consistent with the expected size of the human APJ.The pDsRed-hAPJ recombinant plasmid was cut into two fragments,one corresponded to the pDsRED-express-C1 vector size,and the other fragment corresponded to APJ target fragment.Confocal microscopy analysis showed that,APJ was expressed mainly in the membrane of human embryo kidney 293 cells.The pDeRed-hAPJ eukaryotic plasmid expression vector was successfully constructed and effective expression of this fusion protein is achieved,which might be instrumental in the study of displacement and intracellular localization of human APJ.

Purpose:To study the response and toxicity of the regimen of oxaliplatin combined with fluorouracil and calcium folinate in the treatment of consisting of advanced colorectal cancer.Methods:Thirty-seven patients with advanced colorectal cancer received chemotherapy of regimen:oxaliplatin 130 mg/m~2 2 hours iv on day 1,calcium folinate 200 mg/ m~2 iv 2 hours on days 1 to 5,followed by fluorouracil 300 mg/m~2(≤500 mg/d) iv 4 h on days 1 to 5,three or four weeks as one cycle.Results:The total response rate was 29.7%,the main toxicity was bone marrow suppression and neuro-sensory toxicity,leukopenia was observed in 45.9% of the patients,but grade Ⅲ and Ⅳ in only 8.1%,neuro-sensory toxicity was observed in 81.9.%,but grade Ⅲ and Ⅳ in only 5.4%.Conclusions:This study shows that the regimen of oxaliplatin combined fluorouracil and calcium folinate is effective and tolerable in advanced colorectal cancer therapy.

AIM:To observe the effect of fresh amniotic membrane transplantation ( FAMT) in acute ocular chemical burns.
●METHODS:A prospective study of 25 consecutive cases (36 eyes) with acute ocular chemical burns were treated with FAMT. The clinical efficacy was observed such as the time of amniotic membrane absorbed, corneal epithelialization & transparency, visual acuities and complications.
●RESULTS: With follow-up ranged from 3 to 6mo, 31 eyes′ amniotic membrane were dissolved in 2wk (86%). A total of 33 eyes showed corneal epithelialization in 4wk ( 92%) , 3 eyes showed persistent corneal epithelial defects and need secondary limbal stem cell transplantation or corneal transplantation ( 8%) . A total of 10 eyes showed superficial corneal vascularization (28%), 6 eyes′ cornea were opacity in part (17%), and one eye was symblepharon (3%).
●CONCLUSION:Early FAMT is an effective treatment in the management of acute ocular chemical burns to support epithelial healing, restore ocular surface integrity with potential to improve vision and reduce the incidence of complications. Furthermore, FAMT has advantages of easily obtain and convenient usage, which is suitable in local hospital of our country.

OBJECTIVETo observe the effect of Bushen Wenyang Huayu Recipe (BWHR) on hypoxia inducible factor-1α (HIF-1α), proline hydroxylase2 (PHD2), von Hippel Lindau disease (VHL) suppressor gene expressions in endometriosis (EM) rats with Shen yang deficiency blood stasis syndrome (SYDBSS), and to explore the pathogenesis of EM and the mechanism of BWHR for treating EM.

METHODSTotally 50 SD rats were randomly divided into five groups, i.e., the blank control group, the sham-operation group, the model group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 10 in each group. Rats in the blank control group and the sham-operation group were fed routinely. Rats in the rest 3 groups received 30-day "extended refrigerator freezing and ice water immersion" and combined with " autotransplantation" to establish EM rat model with SYDBSS. One Milliliter BWHR at 3.33 g/mL was administered to rats in the CM group by gastrogavage. Gestrinone at the daily dose of 0. 5 mg/kg was administered to rats in the WM group by gastrogavage. Equal volume of normal saline was administered to rats in the model group, the blank control group, and the sham-operation group. The size and morphology of ectopic foci in rats were observed after 4 weeks of medication. Expressions of serum CA125, plasma cyclic adenosine monophosphate (cAMP), and plasma cyclic guanosine monophosphate (cGMP) were detected by radioimmunoassay. Morphological changes of eutopic endometrium and ectopic tissue were observed under the optical microscope by HE staining. Protein expressions and contents of HIF-lα, PHD2, and VHL were detected by immunohistochemical SABC method and Western blot. mRNA expressions of HIF-1α, PHD2, and VHL were detected by RT-PCR.

RESULTSThe ectopic foci grew significantly in the model group. Their volumes were obviously contracted after treated by CM and WM. Compared with the blank control group and the sham-operation group, serum CA125 and plasma cGMP obviously increased, cAMP obviously decreased (P < 0.05); expressions and contents of HIF-1α mRNA and protein all decreased (P < 0.05); mRNA and protein expressions and contents of PHD2 and VHL all decreased in the model group (P < 0.05). Compared with model group, levels of CA125 and cGMP obviously decreased; cAMP levels obviously increased, expressions and contents of HIF-1α mRNA and protein all increased, mRNA and protein expressions and contents of PHD2 and VHL all increased in the WM group and the CM group (P < 0.05). Compared with the CM group, PHD2 protein contents were higher in the WM group (P < 0.05). HIF-1α was negatively correlated with PHD2 (r = -0.799, P = 0.00). HIF-1α was negatively correlated with VHL (r = -0. 625, P = 0.003).

CONCLUSIONSBWHR could effectively treat EM. Its mechanism might be associated with reducing contents of HIF-1α, serum CA125, and plasma cGMP, and up-regulating expressions of PHD2, VHL, and cAMP.

Animals ; Cyclic AMP ; Drugs, Chinese Herbal ; therapeutic use ; Endometriosis ; drug therapy ; metabolism ; Female ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Proline ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Up-Regulation ; Yang Deficiency ; drug therapy ; metabolism

OBJECTIVETo explore the role of some apoptosis regulators during the development in rat cochlea.

METHODSThe morphological development process of cochlea was observed in Wistar rat aged between embryo day 13 to postnatal day 14 in this experiment. Survivin and caspase-3 were respectively detected at protein and mRNA levels by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe expression of survivin and caspase-3 located in the bottom wall of the cochlear duct. Not only they widespread in the cell proliferation, but also they gradually enhanced in the cell differentiation. Both of them had a crest-time, and survivin was prior to caspase-3. In organ of corti during adult time, caspase-3 was not present and survivin was only expressed.

CONCLUSIONSDuring the development of the rat cochlea, both of them had similar location and trend. But they were derangement. This showed that both of them participated in the cochlear morphological development. It was suggested that the interaction between survivin and caspase-3 regulated the apoptosis, promoted the cochlear morphological development.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Cochlea ; embryology ; growth & development ; metabolism ; Female ; Male ; Microtubule-Associated Proteins ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo investigate the correlation between ER-a in the liver and cytokines of T lymphocytes subsets and serum signatures in PBC patients.

METHODSThe research is performed with cross-sectional study. 80 PBC women patients without treatment were enrolled in PBC group, 10 healthy women as baseline-matched in healthy-control group, and 20 patients with non-autoimmune liver disease in non-PBC control group. The expression of IL-6, IL-8, IL-22, TNFa, IFNgamma, AMA-M2, Sp100 and gp210 were analyzed in Peripheral Blood using ELISA in all groups, and ER-a of patients were performed on tissues from liver biopsies in PBC group and non-PBC control group with immunohistochemistry. Spearman correlation test were performed on the indices to identified the association of all Parameters. numerical data were compared with Wilcoxon rank-sum test.

RESULTSCompared with healthy-control group, expression of serum cytokines are significantly higher in PBC and non-PBC groups (P less than 0.01), while no significant difference were observed between PBC and non-PBC groups. The positive rate of ER-a in PBC patients liver tissues in PBC group is higher than that in non-PBC group (Z=4.82, P less than 0.01). Expression of ER-a is positively correlated with positive rates of AMA-M2 antibody, Sp100 and gP210 of tissues of PBC patients ( r=0.898, 0.819, 0.814, P less than 0.01). ER-a is positive correlated with the expression of cytokines, among which the coefficient of correlation of IL-22, TNFa, IFNgamma is more than 0.7 (r=0.71, 0.89, 0.82, P less than 0.01), AMA-M2, Sp100, gp210 is negative in serum of non-PBC control group. No obviously correlations were indicated between the expression of ER-a and cytokines.

CONCLUSIONA high level of expression of cytokines in the serum might be one of the factors of etiopathogenesis of PBC.

Autoantibodies ; blood ; Biomarkers ; blood ; Case-Control Studies ; Estrogen Receptor alpha ; metabolism ; Female ; Humans ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Interleukins ; blood ; Liver ; metabolism ; Liver Cirrhosis, Biliary ; blood ; immunology ; Middle Aged

BACKGROUNDCentral venous pressure (CVP) and intrathoracic blood volume index (ITBVI) were used to assess the fluid status. It has previously been shown that CVP is not as accurate as ITBVI for all the shock patients. We therefore hypothesized that the change of CVP has the ability to predict fluid responsiveness in some clinical cases of shock.

METHODSFrom September 1st 2009 to September 1st 2011, sixty-three patients with shock from different Intensive Care Unit (ICU) were collected into this retrospective study. All the patients received fluid challenge strategy (infusing 300 ml hydroxyethyl starch in 20 minutes), were monitored with CVP and pulse-indicated continuous cardiac output (PICCO). The correlation between changes in cardiac index (ΔCI), CVP (ΔCVP) and ITBVI (ΔITBVI) were analyzed. Fluid responsiveness was defined as an increase in CI ≥ 10%. Receiver operating characteristic (ROC) curves were generated for ΔCVP and ΔITBVI.

RESULTSFor all the patients, there was no correlation between ΔCI and ΔCVP (P = 0.073), but in the subgroup analysis, the correlation between ΔCI and ΔCVP was significant in those younger than 60 years old (P = 0.018) and those with hypovolemic shock (P = 0.001). The difference of areas under the ROC curves of ΔCVP and ΔITBVI were not statistically significant in the group younger than 60 years old or hypovolemic shock group (P > 0.05, respectively). However, no similar results can be found in the group older than 60 years old and the other two shock type groups from ROC curves of ΔCVP and ΔITBVI.

CONCLUSIONSΔCVP is not suitable for evaluating the volume status of the shock patients with fluid resuscitation regardless of their condition. However, in some ways, ΔCVP have the ability to predict fluid responsiveness in the younger shock patients or in the hypovolemic shock patients.

Adult ; Aged ; Aged, 80 and over ; Central Venous Pressure ; physiology ; Female ; Fluid Therapy ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Shock ; physiopathology ; therapy

OBJECTIVETobacco smoking results in increased platelet aggregability, which suggests that low-dose aspirin used in common clinical practice may not effectively inhibit platelet activity in smokers with coronary heart disease (CHD). This review was performed to assess the effect of aspirin on platelet aggregation in patients with CHD.

DATA SOURCESWe performed an electronic literature search of MEDLINE (starting from the beginning to March 15, 2009) using the term "smoking" or "tobacco" paired with the following: "platelet", "aspirin" or "coronary heart disease".

STUDY SELECTIONWe looked for review articles regarding the effect of tobacco smoking on platelet activity and on the anti-platelet efficacy of aspirin in healthy people and patients with CHD. The search was limited in "core clinical journal". In total, 1321 relevant articles were retrieved, and 36 articles were ultimately cited.

RESULTSTobacco smoking results in increased platelet aggregability, which can be inhibited by low-dose aspirin in the healthy population. However, in patients with CHD, the increased platelet aggregability can not be effectively inhibited by the same low-dose of aspirin. A recent study indicated that clopidogrel or an increased dose of aspirin can effectively inhibit the increased platelet aggregability induced by tobacco smoking in patients with CHD.

CONCLUSIONSIt is important for patients with CHD to quit smoking. For the current smoker, it may be necessary to take larger doses of aspirin than normal or take an adenosine diphosphate receptor inhibitor along with aspirin to effectively inhibit the increased platelet activity.

Aspirin ; therapeutic use ; Coronary Disease ; drug therapy ; Drug Interactions ; Humans ; Platelet Aggregation Inhibitors ; therapeutic use ; Smoking ; adverse effects

The selective removal of low density lipoprotein (LDL) and fibrinogen (Fib) by degraded carrageenan was studied by the present authors. Degraded carrageenan was prepared by acid with carrageenan as the main material. The effects of acid conditions on the molecular weight were investigated, and the proper reaction conditions were ascertained. The results of infrared spectrometry indicated that the degraded carrageenan is a heparin-like polysaccharide. Then the selective removal of LDL/Fibrinogen by degraded carrageenan was studied. When molecular weight was about 10,000, pH was 5.10 and the concentration of degraded carrageenan was 800 mg/L, the average reduction percentages were 60.0% for total cholesterol(TC), 79.4% for LDL and very low-density lipoprotein (VLDL), and 93.8% for fibrinogen. There were no significant changes with relation to the level of high-density lipoprotein (HDL) and total protein (TP). So, degraded carrageenan was shown to be of good selectivity on plasma LDL/Fibrinogen apheresis.
Carrageenan ; chemistry ; Fibrinogen ; analysis ; isolation & purification ; Humans ; Hyperlipidemias ; blood ; Lipoproteins, LDL ; blood ; isolation & purification

Objective To study the variation in numbers of patients attended in the Emergency Department (ER) of a large - scale teaching hospital during weekends or holidays and workdays in order to find out an objective criterion for the assessment of ER overcrowding and the regularity of ER overcrowding.Methods It was a prospective observational study of variation in number of.patient attended in ER during different periods of time round the clock observed from May 1 through October 31 in 2008 -2010 with 110000 emergency patients annually.The roles of diurnal rhythm,holiday phenomenon and medical coverage in the variation in numbers of patients were observed.The multiple logistic regression analysis was used to define the criterion of ED overcrowding.Results During workdays,the regularity of variation in number of critically ill patients seen to in ER was distinctive,the number of patients peaked in the period of 20:00 -22:00 and bottomed out in the period of 4:00 -6:00,while overcrowding scores of both peak and bottom were carried out 2 hours later.The number of emergency patients significantly increased at weekends and long holidays in a form of double peaks,from 10 am to 12 pm and 8 pm to 10 pm.The number of emergency patients was obviously determined by the provisions of medical coverage,but it was only true to non - critical patients,while the number of critical patients did not noticeably change during weekends or holydays.Multivariate regression analysis showed that the number of emergency patient attended in ER ( B =0.027,P ＜0.01 ) and the rate of emergency bed occupancy ( B =5.25,P ＜0.01 ) in the period of two hours significantly correlated with the ER overcrowding in the coming period of two hours (B =0.027,P ＜0.01,B =5.25,P ＜ 0.01,respectively).Conclusions The demand for critical care resources varies up and down all the time.The variation in volume of critical patients is quite regular during workdays and weekdays or holydays.It is important to separate critical patients from non - critical patients in order to divert non - critical patients quickly.Prediction of overcrowding in ER can be made with knowledge of the number of patient attended and the rate of bed occupancy,if the provisions of medical coverage unchanged.This regularity of variation in number of patients can be used as a practical guidance to rational allocation of critical care resources and improvement of patient throughput.