The DNA fragment sized 2 139bp, the same Sequence with AtKup1 gene from Arabidopsis thalianna was used as the templates for DNA family shuffling. The shuffeld AtKup1 gene library was expressed in the mutant of 5. cerevisae in which potassium transporter gene TRK1 and TRK2 were knocked out by homologous recombination. Then the screening was carried out in the low potassium media containing 5. 0 mmol/L KC1 and no histidine in it. it was found that both of diverse and wild AtKup1 gene can rescues the trk1△trk2△yeast mutant strain in low [ K + ] medium. The growth of 2 clones yeast containing diverse AtKup1 were beter than that of AtKup1 wild gene transformant. The sequencig results of the shuffeld AtKup1 showed that there were 2 nucleotide changed, which resulted in 2 amino acid variations in it compared with the original AtKup1. The potassium uptaking capacity of shuffled AtKup1 gene increased significantly when it was transformed into tobacco.

To study the effect of Yinghua Pinggan granule (YHPG) against influenza A/H1N1 virus in vivo and on the immunologic function of infected mice. The intranasal influenza virus infection was adopted in ICR mouse to establish the influenza virus pneumonia model. At the 3rd and 7th day after the infection, the lung index and pathologic changes in lung tissues of mice were detected. Realtime PCR and flow cytometry were employed to observe the virus load in lung tissues and the levels of CD4+, CD8+, and CD4+/CD8+ in peripheral blood. The result showed that at the 3rd and 7th day after the infection, YHPG (15, 30 g x kg(-1)) can significant decrease in the lung index and virus load in lung tissues of mice infected with influenza virus, alleviate the pathologic changes in lung tissues, significantly increase the levels of CD4+ and CD4+/CD8+ ratio and reduce the levels of CD8+ in whole blood. This indicated that YHPG can inhibit the influenza virus replication, alleviate pulmonary damage and adjust the weak immunologic function of infected mice, with a certain therapeutic effect on mice infected by H1N1 virus in vivo.
Animals ; Antiviral Agents ; administration & dosage ; Humans ; Influenza A Virus, H1N1 Subtype ; drug effects ; genetics ; physiology ; Influenza, Human ; drug therapy ; pathology ; virology ; Lung ; pathology ; virology ; Male ; Mice ; Mice, Inbred ICR ; Virus Replication ; drug effects

OBJECTIVETo discuss the diagnostic value of an ultrasonic assessing system for detecting the severity of hepatic fibrosis in patients with chronic hepatitis B (CHB).

METHODSUltrasonographic variables were analyzed in 110 CHB patients. An ultrasonic semi-quantitative scoring system using seven ultrasonic morphologic parameters, a Fisher discriminating function and three quantitative ultrasonic parameters was developed. The performance of these methods was also studied and compared.

RESULTSThe areas under the curve of the scoring system for different liver fibrosis stages were >or= S2: 0.946, >or= S3: 0.914, and S4: 0.915. The total score was well correlated with the histological stage of fibrosis (r=0.824, P < 0.001). There was a significant difference between the stages of fibrosis. The accuracy of the Fisher discriminating function for identifying three study endpoints was 76.5%, 78.2% and 67.3%. Combining the ultrasonic scoring system and the discriminating function, the specificity was 85%-90% and the accuracy was 77%-84%.

CONCLUSIONOur ultrasonic semi-quantitative scoring system is a noninvasive method for quantitating liver fibrosis. If it is used together with a discriminating function, the accuracy of diagnosing liver fibrosis can be significantly increased.

Adolescent ; Adult ; Aged ; Female ; Hepatitis B, Chronic ; diagnostic imaging ; Humans ; Liver Cirrhosis ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Ultrasonography, Doppler, Color ; Young Adult

OBJECTIVETo establish of acute respiratory distress syndrome (ARDS) model in canine after inhalation of perfluoroisobutylene (PFIB), and to observe the progressing of lung injury, and to study the mechanisms of injury.

METHODSA device of inhalation of PFIB for canine was made. The concentration of PFIB was 0.30 - 0.32 mg/L. Serum IL-6 and IL-8 were dynamically measured. Clinical manifestations, pathology of organs in canine were observed.

RESULTS(1) During inhalation, the concentration of PFIB remained stable; (2) After inhalation, blood arterial oxygen partial pressure fell gradually, and eventually met the criteria for diagnosing ARDS; (3) The level of IL-8 in serum rises significantly after inhalation (P < 0.05), whereas that of IL-6 was not obviously altered (P > 0.05); (4) Within 6 hours after inhalation, no abnormality in canine was observed, but afterwards symptoms gradually appeared, and typical breath of ARDS, such as high frequency and lower level could be seen in later phase; (5) Pathological examination showed severe congestion, edema and atelectasis in most part of both lungs, and signs of anoxia in other organs.

CONCLUSIONS(1) The device designed is capable of ensuring control of inhalation of PFIB; (2) Exposure to PFIB for 30 mins, canines all met the criteria for diagnosing ARDS 22 hours after inhalation, therefore the modeling is successful; (3) PFIB specifically damages the lung by causing excessive inflammation.

Administration, Inhalation ; Animals ; Disease Models, Animal ; Dogs ; Female ; Fluorocarbons ; toxicity ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Lung ; drug effects ; pathology ; Male ; Random Allocation ; Respiratory Distress Syndrome, Adult ; blood ; chemically induced

AIMTo screen swimming-fatigue related genes in mice and lay theoretic basis for researching the molecular mechanism of fatigue.

METHODS30 male BALB/c mice (20 +/- 2g) were divided into control group, dipping in water group and swimming-fatigue group respectively. After fatigue for swimming in swimming-fatigue group, with control group and dipping in water group, liver tissues in mice were collected. With improved silver staining mRNA differential display method, the differentially expressed genes in mice livers were screened and evaluated by reversed Northern blot. The positive segments were analyzed homology by BLAST.

RESULTS7 of DD-ESTs were gained. Two of them only expressed in swimming-fatigue group, two down-regulated expressed, and three up-regulated. One of them was a novel gene and was accepted by GenBank, AY615302.

CONCLUSIONSeven DD-ESTs in swimming-fatigue mice were gained by silver staining mRNA differential display method.

Animals ; Fatigue ; metabolism ; Gene Expression Profiling ; methods ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; RNA, Messenger ; genetics ; Silver Staining ; Swimming

Objective: To establish an echocardiography parameter scoring system for assessing the risk of 1 year re-admission in patients with left ventricular systolic dysfunction (LVSD). Methods: A total of 412 chronic LVSD patients treated in our hospital from 2007-01 to 2016-01 were studied and the end point event was 1 year re-admission. The data included in 280 patients from 2007-01 to 2014-12 for establishing the scoring system and 132 patients from 2015-01 to 2016-01 for verifying the system. Based on 7 echocardiography parameters, the patients were divided into 7 sets of groups: ① Left ventricular diameter (LVD): Group0, n=290 and Group1, n=122;② Mitrial regurgitation (MR): Group0, n=203, Group1, n=138 and Group2, n=71; ③ Tricuspid regurgitation (TR): Group0, n=302, Group1, n=90 and Group2, n=20; ④ LVEF: Group0, n=272 and Group1, n=140; ⑤ Pulmonary artery systolic pressure: Group0, n=282 and Group1, n=130; ⑥ Hydropericardium: Group0, n=347 and Group1, n=65; ⑦ Hydrothorax:Group 0, n=261, Group1, n=86 and Group2, n=65. The parameters were identified by COX regression analysis, weighted value of scoring system was calculate by hazard ratio (HR), predictive value for1 year re-admission was assess by ROC curve and finally, scoring integration was verified by validation data group. Results: The integration score was calculated as follows: LVD>60mm=1 point; TR: Group1=1 point and Group2=3 points; MR: Group1=2 points and Group2=4 points; Hydrothorax: Group1=2 points and Group2=3 points;Hydropericardium=1 point. COX regression analysis indicated that for 1 year re-admission: HR=1.552 in Group1 vs Group0, HR=3.374 in Group2 vs Group0 and HR=4.562 in Group3 vs Group0, all P<0.05. The AUC of ROC for establishing the data was 70.0% (95% CI 0.640-0.761) and for verifying the data was 70.4% (95% CI 0.616-0.792); the best integration score was 4 points. Conclusion: Echocardiography parameter scoring system may better predict the risk of 1 year re-admission in LVSD patients which is superior to single echocardiography parameter.

ObjectiveTo investigate the clinical features of systemic lupus erythematosus (SLE) patients with fever and find out the related factors.MethodsData was collected by the same methods in the past ten years in fifteen hospitals in Jiangsu province and then the data wereretrospectively analyzed.The potentially possible risk factors of fever in SLE were selected and then analyzed by chi-square test,Wilcoxon rank sum test and Logistic regression analysis.ResultsAll 1762 patients were investigated.Seven hundred and twenty-nine had active fever.Age at hospitalization,initially treated patients,photosensitivity,serositis,nervous system involvement,generalized lymphadenopathy/hepatosplenomegaly,white blood cell count (WBC),haemoglobin (HB),erythrocyte sedimentation rate (ESR),C-reaction protein (CRP),alanine aminotransferase(ALT),albumin(ALB),serum creatinine (Scr),complement C3,anti-dsDNA antibodies positive rate,anti-Sm antibodies positive rate,SLEDAI score and past therapies were factors associatedwith SLE fever.Logistic regression analysis showed that abnormal WBC count (OR=1.396,95％CI 1.114-1.711,P=0.004),CRP(OR=1.005,95％CI 1.002-1.009,P=0.002),ALT(OR=1.003,95％CI 1.001-1.005,P=0.005),Scr (OR=0.997,95％CI0.995-0.999,P=0.007),HB (OR=0.986,95％CI 0.981-0.992,P=0.000),age (OR =0.984,95％ CI 0.974-0.993,P=0.001 ) and past usage of cyclophosphamide (CTX) (OR =0.557,95％CI 0.382-0.813,P=0.002) were correlated with SLE fever.ConclusionFever is one of the most common clinical manifestations of SLE patients.Leucopenia,elevated CRP levels,liver function abnormalities,anemia,younger age are risk factors for SLE fever,while renal impairment and past usage of CTX are protective factors.

OBJECTIVETo analyse the clinical feature and natural course of essential thrombocythemia (ET).

METHODSA retrospective analysis was conducted in ET patients treated in our hospital during May 1980 to December 2006.

RESULTSFour hundred and thirty eight patients (201 males and 237 females with a median age of 48 years) were diagnosed. Hemorrhage occurred in 101 cases (23.1%), thrombosis in 86 cases (19.6%), and both hemorrhage and thrombosis in 13 cases (3.0%). Splenomegaly occurred in 150 cases and hepatomegaly occurred in 60 cases. One hundred and forty-nine cases (34%) had no symptoms at diagnosis and 145 cases (33.1%) confirmed by routine blood tests due to other diseases. The median platelet count at diagnosis was 1000 x 10(9)/L [(533 -3740) x 10(9)/L]. Bone marrow biopsy was performed in 255 cases which showed mainly increase of enlarged mature megakaryocytes with hyper-lobulated nuclei and local proliferation of reticular fiber was revealed in 51 cases. JAK2V617F mutation was detected in 90(78.9%) of 114 patients studied. Karyotype analysis was performed in 180 cases and 6 (3.3%) had clonal chromosomal aberrations. Two hundred and sixty-one patients were followed up over 12 months with a median of 60 months (range from 12 to 300 months). Seventeen cases (6.5%) evolved into marrow fibrosis (MF) and one case into polycythemia vera (PV). One case evolved into PV 6 years and then MF 20 years after diagnosis of ET. Three cases developed acute monocyte leukemia (M5), myelodysplastic syndrome (MDS) and multiple myeloma (MM), respectively.

CONCLUSIONSET is a chronic myeloproliferative disorder characterized predominantly by thrombocytosis and hemorrhage. The percentage of asymptomatic cases is high. The prognoses for most cases were good with a few cases may evolve into MF.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Thrombocythemia, Essential ; genetics ; pathology ; Young Adult

OBJECTIVETo detect absorbed bioactive compounds of the water extract whose pharmacodynamic effect was craniocerebral protection for quality control assessment.

METHODSAnthraquinones in water extract of rhubarb (WER), in cerebrospinal fluid (CSF) of patients with traumatic brain injury (TBI) and in ipsilateral cortex of TBI rats following oral WER were respectively explored by ultra performance liquid chromatography with photodiode array detector (UPLC-PDA) method developed in the present study. The effects of anthraquinones absorbed into injured cortex on superoxidase dismutase (SOD) activity in TBI rats were detected. The antioxidative anthraquinones absorbed into target organ were evaluated for quality control of WER.

RESULTSAnthraquinones in WER were aloe-emodin, rhein, emodin, chrysophanol, and physcion. Only the last anthraquinone was found in CSF and in ipsilateral cortex under this chromatographic condition. Physcion increased SOD activity in TBI rats significantly.

CONCLUSIONSPhyscion was the main active compound of rhubarb against craniocerebral injury via antioxidant pathway. According to our strategy, the exploration of physcion suggested the possibility of a novel quality control of WER in treating TBI injury.

Absorption ; drug effects ; Animals ; Anthraquinones ; cerebrospinal fluid ; chemistry ; Biological Products ; analysis ; cerebrospinal fluid ; chemistry ; Brain Injuries ; drug therapy ; pathology ; Chromatography, Liquid ; instrumentation ; methods ; Emodin ; administration & dosage ; analogs & derivatives ; pharmacology ; therapeutic use ; Humans ; Limit of Detection ; Linear Models ; Male ; Plant Extracts ; chemistry ; Quality Control ; Rats ; Rats, Sprague-Dawley ; Reference Standards ; Reproducibility of Results ; Rheum ; chemistry ; Water ; chemistry

OBJECTIVETo investigate the effect of chronic enhanced external counterpulastion (EECP) on gene expression profiles of arterial endothelial cells (ECs) of pigs fed with high-cholesterol diet.

METHODSEight male pigs were fed with high-cholesterol diet for 12 weeks to induce arteriosclerosis and subjected to EECP for accumulative 36 h (2 h every other day for 18 sessions). Another 8 pigs on cholesterol-enriched diet and 6 normally fed pigs served as the arteriosclerosis model group and normal control group, respectively, and the high-cholesterol diet was maintained until the end of EECP treatment. The coronary artery was then isolated for transmission electro microscopy, and the abdominal aorta was observed using Sudan III staining. The gene expression profiles in ECs from the thoracic aorta using cDNA microarrays.

RESULTSMacrophages and foam cells were detected beneath the ECs in the coronary artery of pigs in the model group, but not in the other two groups. The ratios of Sudan III-positive area in the celiac aorta were significantly lower in normal control and EECP groups than in the model control group (P<0.05). Compared with the normal control group, the gene expressions of integrins-beta1 and CTGF were up-regulated in the model group. Compared with the model group, the expressions of integrins-beta1, CTGF and VCAM-1 were down-regulated and eNOS up-regulated in EECP group.

CONCLUSIONChronic EECP may reduce endothelial injury, down-regulate the gene expression level of integrin-beta1, CTGF and VCAM-1, lower cholesterol uptake and attenuate arterial endothelial inflammation to protect the pigs fed with high-cholesterol diet from arteriosclerosis.

Animals ; Aorta, Abdominal ; metabolism ; pathology ; Arteriosclerosis ; etiology ; genetics ; pathology ; Coronary Vessels ; metabolism ; pathology ; Counterpulsation ; methods ; Diet, Atherogenic ; Endothelial Cells ; metabolism ; Gene Expression Profiling ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Swine