Objective To study the relationship between long-term high-salt water and the morbidity mechanism of chronic atrophic gastritis (CAG), The expression of HSP60 and ras protein were detected in the gastric mucosa of CAG rates induced by high-salt water.Methods The atrophic gastritis rat model was made by high-salt-hot water perfusion and the ultrastructure of gastric mucosa was observed by Laser Scanning ConfocaI Microscopy. Results There was no expression in gastric mucosa in normal group. The expression of HSP60 and ras was observed in the cell plasm of rats at 12 weeks. The higher expression was observed in the rats at 24, 32 and 65 weeks. Using laser scanning confocaI microscopy and immunofluorescence technique had observed coexistence of HSP60 and ras.Conclusion Long time high-salt-hot water can induce CAG and increasing the expression of HSP60 and ras. HSP60 and ras play an important role in the formation of atrophic gastritis.

Objective To explore the curative effect of tirofiban treatment on high-risk acute coronary syndromes (ACS) in elderly patients receiving an early percutaneous coronary intervention (PCI) treatment. Methods The 162 elderly cases including unstable angina pectoris and non-ST -segment elevation myocardial infarction (NSTEMI) undergoing early PCI were enrolled in this study.And they were assigned to early treatment group (n=82) and deferred selective group (n=80)according to the time of using tirofiban (Gp Ⅱ b/Ⅲ a inhibitor) treatment. The effectiveness of either strategic option on tissue-level perfusion was evaluated using the TIMI myocardial perfusion grade (TMPG) before and immediately after PCI. The corrected TIMI frame count (cTFC) was also used to assess coronary artery flow and myocardial perfusion. Bleeding complications and the composite end point events at 30 days were also evaluated. Results Of all the 162 patients, the TMPG 0-1 perfusion was observed in 65 patients (40.1%). The TMPG 0-1 perfusion was significantly less frequent in early treatment group (32.9%) than in deferred selective group (47.5%) before PCI (x2=3.58, P＜0.05); while the results of TIMI grade 0-1 flow (26.8% vs. 25.0%) and cTFC levels (34.2±11.8 vs. 34. 9±12. 7) before PCI were similar between the two groups (x2 =0. 07, P=0.47; t= 0.13, P=0.71, respectively). No differences were seen both in composite end point events at 30 days and bleeding complications (x2 = 0.31, P＞0.05; x2=0.004, P＞0. 05). Conclusions High -risk ACS patients treated with an early invasive strategy, routine upstream use of tirofiban are associated with improved tissue-level perfusion before PCI and does not increase bleeding complications when bleeding risks are carefully evaluated before enrollment.

Obej ctive Knockout mice are widely used in the studies of joint diseases .This article investigated the effects of joint processing methods , collagenase types ,and collagenase digestion time on the number of primary fibroblast -like synoviocytes (FLSs) obtained from mice. Methods The hind legs of 6 of the 12 male mice were cut open from the hip joints , but not those of the other 6.FLSs were isolated using the type-Ⅳcollagenase digestion method and purified by differential digestion .Cell morphology was observed under the inverted microscope .The type, viability, and purity of the cells were determined by flow cytometry . Rse ults Significantly fewer FLSs were obtained from the mice with the hind legs cut open ( 19 133 ±115 ) than from those without (24 933 ± 503) (P<0.05).The numbers of FLSs collected from the cell suspension at 1, 2, 3, 4, 5,6 , and 7 hours after digestion were 700 ±300 , 600 ±100 , 15 200 ±900 , 5100 ±800 ,2700 ±300 , 900 ±200, and 300 ±100, respectively, the highest at 3 hours. There were statistically significant differences in the total number of FLSs obtained by type-Ⅳ and type-Ⅱ collagenase digestions (24900 ±500v s 18 100 ±400, P<0.05). Conclusion For in virt o culture of primary mouse FLSs, it is recommended that the hip joints be not cut open, and type-Ⅳcollagenase be used with cell sus-pension at 2-6hours after digestion .

Objective To investigate the sleep disorders status of patients with peritoneal dialysis (PD),and analyze the related factors of sleep disorders.Methods The KDQOL sleep sub-scale was used to conducted the sleep investigation in 84 PD patients.Meanwhile,the fatigue,physical activity level and depression were collected.Results The mean score of sleep sub - scale of patients was (53.12 ± 19.33 ).The prevalence of sleep disorders was 48.8％.And the logistic regression analysis showed that low physical activity level( OR =2.172,P ＜0.05 ),medical expenses ( OR =1.793,P ＜0.05 ) and depression ( OR =1.367,P ＜0.05) were predicted factors of sleep disorders.Conclusions More attention should be paid to sleep disorders in PD patients,promote them to take part in physical activity,take medicine or non-medicine measures to improve their quality of sleep and mental health so as to decrease the incidence of sleep disorders in PD patients.

BACKGROUNDMutations in fumarylacetoacetate hydrolase (FAH) gene can lead to tyrosinemia type 1 (HT1), a relatively rare autosomal recessive disorder. To date, no molecular genetic defects of HT1 in China have been described. We investigated a Chinese family with a HT1 child to identify mutations in FAH.

METHODSDNA sequencing was used for mutations screening in FAH gene. Real-time polymerase chain reaction (PCR) was performed to determine the FAH gene expression level. To confirm the presence of degradation by the nonsense-mediated mRNA decay pathway (NMD), the fragments containing R237X mutations were analyzed by primer introduced restriction analysis-polymerase chain reaction (PIRA-PCR) and cDNA sequencing. Finally, the effects of the mutations reported in this study were predicted by online softwares.

RESULTSA boy aged 3 years and 8 months was diagnosed clinically with HT1 based on his manifestations and biochemical abnormalities. Screening of FAH gene revealed two heterozygous mutations R237X and L375P transmitted from his mother and father respectively. In this pedigree, the amount of FAH mRNA relative to a healthy control was 0.44 for the patient, 0.77 for his mother and 1.07 for his father. Moreover, both PIRA-PCR and cDNA sequencing showed significant reduction of the FAH mRNA with R237X nonsense mutation. The missense mutation of L375P was not reported previously and prediction software showed that this mutation decreased the stability of protein structure and affected protein function.

CONCLUSIONSThis is the first case of HT1 analyzed by molecular genetics in China. The R237X mutation in FAH down- regulates the FAH gene expression, and the L375P mutation perhaps interrupts the secondary structure of FAH protein.

Child, Preschool ; China ; Humans ; Hydrolases ; genetics ; Male ; Molecular Sequence Data ; Mutation ; Mutation, Missense ; genetics ; Nonsense Mediated mRNA Decay ; genetics ; Real-Time Polymerase Chain Reaction ; Tyrosinemias ; genetics

The gene (eg1) encoding for novel endoglucanase 1 was cloned previously from Chinese straw mushroom Volvariella volvacea. EG1 has high thermal stability and optimal pH at neutral and shows great potential in textile and paper industry applications. To improve the expression level of EG1 in Pichia pastoris, the increasing copy number of clone, and its high cell density fermentation in 3.2L fermenter for its high-level expression were investigated in this work. By electro-transformation of pPICZalphaB-egl into GS115EG11 integrated with single copy of eg1 gene, A resistant transformant with 3.8 times higher level expression than GS115EG11 was screened from YPDSZ plate containing 2000 microg/mL of Zeocin. The effect of initial cell density, pH and methanol on its expression and biomass accumulation was evaluated in shaking culture. Optimal EG1 production was observed when initial cell density OD600 was 5.0. EG1 production and biomass accumulation did not seem to vary when cells were induced at different pH values. Both of EG1 and cell density were found to increase with higher methanol concentrations, reaching 62.48 IU/mL and 31.7 (OD600) respectively after 120 h induction with 2.0% (V/V) methanol compared to 30.24 IU/mL and 17.79 (OD60) with 0.25% methanol induction. EG1 expression was further increased by 6.4 times higher than shaking culture after 95.5 hours induction with methanol in fed-batch fermentation, so totally 34 times higher than that for GS115EG11 was achieved by screening of high Zeocin resistant clone and high cell density fermentation. The production of EG1 with 543.36IU/mL CMC activity and 8.80mg/mL protein expression was obtained in Pichia pastoris.
Cellulase ; biosynthesis ; genetics ; Cloning, Molecular ; Culture Media ; Electroporation ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Fungal Proteins ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Volvariella ; enzymology ; genetics

AIMTo investigate whether mitochondrial calcium uniporter participates in the cardioprotection of tumor necrosis factor alpha (TNFalpha) pretreatment in isolated rat hearts subjected to ischemia/reperfusion.

METHODSIsolated perfused rat hearts were subjected to 30 min regional ischemia (occlusion of left anterior descending artery) and 120 min reperfusion. The infarct size, coronary flow (CF) and lactate dehydrogenase (LDH) release during reperfusion were measured. The mitochondria of the heart were isolated and suspended in the swelling buffer for measurement of absorbance at 520 nm.

RESULTSPretreatment with TNFa at 10 U/ml for 7 min followed by 10 min washout reduced the infarct size and LDH release, and improved the recovery of CF during reperfusion. Administration of spermine (20 micromol/L), an opener of mitochondrial calcium uniporter, for 10 min during early reperfusion attenuated the reduction of infarct size and LDH release, and improvement of CF induced by TNFalpha. In isolated mitochondria of the heart pretreated with TNFalpha, the absorbance at 520 nm decreased less than that of mitochondria without TNFalpha pretreatment. Administration of spermine (50 micromol/L) attenuated the change of the absorbance induced by TNFalpha.

CONCLUSIONThe findings indicate that TNFalpha protects myocardium against ischemia/reperfusion injury via inhibiting mitochondrial calcium uniporter opening as well as mitochondrial permeability transition pore opening.

Animals ; Calcium Channels ; drug effects ; metabolism ; Cardiotonic Agents ; pharmacology ; In Vitro Techniques ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Mitochondrial Membrane Transport Proteins ; drug effects ; Myocardial Reperfusion Injury ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Spermine ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology

The aim of this study is to investigate the anti-inflammatory effect of the adenosine derivative N6-(3-hydroxylaniline) adenosine (WS070117M1) on cigarette smoke plus LPS (lipopolysaccharide)-induced chronic obstructive pulmonary disease (COPD) in mice and its mechanism. COPD model was established by exposing male BALB/c mice to cigarette smoke and challenged with LPS inhalation. Supernatants of bronchoalveolar lavage fluid (BALF) were harvested and IL-1β, IL-6, IL-8 and TGF-β1 levels were measured by ELISA (enzyme-linked immunesorbent assay). The number of total white blood cells and neutrophils in bronchoalveolar lavage fluid was counted separately. Lung tissue was stained with Mayer 's hematoxylin and eosin for histopathologic examination. pAMPKa protein expression and distribution of lung tissue were analyzed by immunohistochemistry method. In vitro, levels of AMPKα phosphorylation in phorbol-12- myristate-13-acetate (PMA) differentiated THP-1 cells was detected by immunohistochemistry, IL-8 level in supernatants of cigarette smoke condensate stimulating PMA differentiated THP-1 cells was measured by ELISA. The results showed that WS070117M1 treatment significantly activated AMPKa in the lung tissue. It also resulted in down regulation of IL-1β, IL-6, IL-8 and TGF-β1 levels in bronchoalveolar lavage fluid and IL-8 level in cigarette smoke condensate stimulating PMA differentiated THP-1 cells. In addition, WS070117M1 could inhibit the recruitment of total white blood cells and neutrophils. These results suggest that WS070117M1 may alleviate the airway inflammation by activating AMPK in the lung tissue.
AMP-Activated Protein Kinases ; metabolism ; Adenosine ; analogs & derivatives ; Animals ; Bronchoalveolar Lavage Fluid ; Cell Line, Tumor ; Disease Models, Animal ; Humans ; Inflammation ; drug therapy ; Interleukin-1beta ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Leukocyte Count ; Lipopolysaccharides ; Male ; Mice ; Mice, Inbred BALB C ; Neutrophils ; cytology ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; Smoke ; adverse effects ; Tobacco ; Transforming Growth Factor beta1 ; metabolism

Objective To explore the application of standardized patients (SP) in practice course of diagnostics.Methods The two classes of 135 students majoring in clinical medicine were selected as research.After the practical course in inquiry,the questionnaires were used to compare the teaching effect between the traditional teaching and SP teaching.Meanwhile,the students evaluated and gave feedback on SP work.Results The questionnaires on teaching effect showed that 127 (94.8％) students believed that the SP teaching method could stimulate interest in inquiry,125 students (93.3％) thought it could increase the understanding and memory of the inquiry content.127 students (94.8％) thought it could improve the application ability of the inquiry method,128 students (95.5％) thought it could improve inquiry techniques,129 students (96.3％) thought it could enhance expression ability,89 students (66.4％) thought it could enhance cooperation with their classmates,117 students (87.3％) thought it could enhance the confidence in communication,115 students (85.8％) thought it could improve their ability in independent thinking and self-study,126 students (94.0％) thought it could improve the learning effect,117 (87.3％) students thought it could develop their clinical thinking ability,116 students (86.6％) thought it could improve the satisfaction to teaching form,111 students (82.8％) thought it could develop the ability to answer patients' problems in a manner which is easy for patients to understand,and 126 (94.0％) students thought it could improve the ability to interact with patients.All issues were significantly better than the traditional teaching method Compared with the traditional teaching method,the students' choices on various items have statistically significant differences (P＜0.05).Conclusion Applying SP teaching to the practice courses of diagnostics is helpful to improve the teaching effect and cultivate students' clinical thinking ability.Thus,it is a promising teaching method in medical undergraduate education and worthy of further discussion and popularization.

Objective The aim of this study was to introduce the multi-slate Markov model for the prediction of mild cognitive impairment (MCI) to Alzheimer' s disease (AD) and to find out the related factors for AD prevention and early intervention among the elderly.Methods MCI,moderate to severe cognitive impairment,and AD were defined as state 1,2 and 3,respectively.A three-state homogeneous model with discrete states and discrete times from data on six follow-up visits was constructed to explore factors for various progressive stages from MCI to AD.Transition probability and survival curve were made after the model fit assessment.Results At the level of 0.05,data from the multivariate analysis showed that gender (HR=I.23,95％CI:1.12-1.38),age (HR=I.37,95％ CI:1.07-1.72),hypertension (HR=l.54,95％ CI:1.31-2.19) were statistically significant for the transition from state 1 to state 2,while age (HR=0.78,95％ CI:0.69-0.98),education level (HR=1.35,95％ CI:1.09-1.86) and reading (HR=1.20,95％ CI:1.01-1.41 ) were statistically significant for transition from state 2 to state 1,and gender (HR=1.59,95％ CI:1.33-1.89),age (HR=1.33,95％ CI:1.02-1.64),hypertension (HR=l.22,95％ CI:1.11-1.43),diabetes (HR=1.52,95％CI:1.12-2.00),ApoEε4 (HR=1.44,95％CI:1.09-1.68) were statistically significant for transition from state 2 to state 3.Based on the fired model,the three-year transition probabilities during each state at average covariate level were estimated.Conclusion To delay the disease progression of MCI,phase by phase prevention measures could be adopted based on the main factors of each stage.Multi-state Markov model could imitate the natural history of disease and showed great advantage in dynamically evaluating the development of chronic diseases with multi-states and multi-faetors.