Objective:To explore the reliability and validity of the Chinese version of Repeatable Battery for the Assessment of Neuropsychological Status(RBANS),which is specially for assessing cognitive functions in non-dementia mental disorders.Methods: Altogether 451 healthy adults who were recruited from both rural and urban areas in Beijing were evaluated with the RBANS,and 97 adults were tested with the Brief Wechsler Adult Intelligence Scale(WAIS) and Wechsler Memory Scale simultaneously(WMS).Forty-one adults were reevaluated with RBANS 12 weeks later.The data was analyzed by using correlation analysis and factor analysis.Results: The Cronbachs'? coefficient of RBANS total scale was 0.90;the Cronbachs'? coefficients of immediate memory,visuospatial,language,attention and delayed memory were 0.86,0.68,0.67,0.85 and 0.80 respectively.The test-retest reliability of total scale was 0.90 and that of subscales were 0.65,0.68,0.53,0.80 and 0.79 respectively(P

Adult ; Antineoplastic Agents ; toxicity ; CD4-Positive T-Lymphocytes ; drug effects ; Female ; Humans ; Killer Cells, Natural ; drug effects ; Middle Aged ; Nurses ; statistics & numerical data ; Occupational Exposure ; T-Lymphocyte Subsets ; drug effects

Adolescent ; Adult ; Aged ; Endoscopy ; Female ; Humans ; Male ; Maxillary Sinus ; surgery ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; methods ; Young Adult

Aim:To construct the short hairpin small interfering RNA(shRNA) eukaryotic expression vector specific to mouse lectin like oxidized low density lipoprotein receptor 1(LOX-1) gene and to observe its silencing effect on LOX-1 in RAW264.7 cells.Methods:(1)The pLOX-1-shRNA expression vector was constructed by gene recombination,Then transfected into the cultured RAW264.7 cells.At 48 h after Transfection,the expression of LOX-1 mRNA in RAW264.7 cells were determined by semi-quantitative RT-PCR,the expression of LOX-1 proteins examined by Western blot.(2) Oil Red O Dyeing experiment was used to show the cellular lipid droplets in lipid-loaded cells.The method of cholesterol oxidase analysis was performed to determine the content of cellular cholesterol.The ability of uptake Dil-ox-LDL in RAW264.7 cells was assayed by fluorescence microscopy.Results: pLOX-1-shRNA expression vector was successfully constructed.Transfection of pLOX-1-shRNA expression vector into RAW264.7 cells down regulaled the expression level of LOX-1 gene,as compared with the control group,transfection of the RAW264.7 cells with LOX 1-shRNA led to a remarkable reduction of the number macrophages transformed into foam cell,and could suppress the uptake of ox-LDL.Conclusion:The pLOX-1-shRNA expression vector can inhibit the expression of LOX 1 in RAW264.7 cells and the transformation of the macrophages into foam,which may he beneficial in searching new gene therapy of atherosclerosis.

The uracil in DNA comes from either the misincorporation of dUTP in place of dTTP or deamination of cytosine. In the latter case, it can result in a GC to AT transition mutation if the uracil is not removed before DNA replication. Base excision repair (BER) is a major pathway for removing DNA lesions arising from endogenous processes as well as those induced by exposure to exogenous chemicals or irradiation. BER is initiated by DNA glycosylases that excise aberrant bases from DNA by cleavage of the N-glycosidic bond linking to the base of its deoxyribose sugar. Uracil N-glycosylase (UNG) is the enzyme responsible for the first step in the BER pathway that specifically removes uracil from DNA. The UNG gene undergoes both temporal and spatial regulation mainly at the level of transcription. Normally cancer cells undergo over-proliferation and up-regulate their UNG during tumorigenesis. In this study we examine the correlation between UNG level and carcinogenesis, and explore the possibility of using UNG as a marker for cancer diagnosis. Human UNG gene was amplified from the total RNA of the human choriocarcinoma cell line, JEG-3, by RT-PCR. After purification, the 942bp full-length UNG cDNA coding sequence was digested with EcoR I and Sal I, and cloned into the digested pET-21 to construct a recombinant vector, pUNG. The UNG protein was expressed under the control of T7 promoter in E. coli BL21 (DE3) cells induced with IPTG. After ultrasonic treatment, the cell lysate and precipitate were analyzed by SDS-PAGE and a 39kD band was detected. The plasmid was serially diluted at appropriate concentrations and employed as standards in the subsequent quantification. Total RNAs were extracted from 18 pairs of clinical samples, each pair contains a sample of esophageal squamous cell carcinoma (ESCC) tissue and its surrounding normal esophageal epithelia. The copy numbers of UNG mRNA in these RNA samples were determined by real-time quantitative RT-PCR using a Lightcycler (Roche). UNG was present in 13 cases of ESCC (13/18, n = 18) but absent in all of the normal tissues. The results indicated that there was a correlation between high level of UNG expression and the carcinogenesis of ESCC.
Carcinoma, Squamous Cell ; genetics ; metabolism ; Cell Line, Tumor ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Esophageal Neoplasms ; genetics ; metabolism ; Humans ; In Vitro Techniques ; Reverse Transcriptase Polymerase Chain Reaction ; Uracil-DNA Glycosidase ; genetics ; metabolism

Chaetomium thermophilum CT2 can produce extracellular cellulase with industrial value. We designed two degenerate primers to amplify catalytic domain sequence of cellobiohydrolase II ( CBH II). Full length of cDNA was obtained by rapid amplification of cDNA ends technologies. DNA sequencing revealed that cbh2 has an open reading frame of 1428bp, which encodes a putative polypeptide of 476 amino acids. The deduced amino acid sequence shows that the predicted molecular mass is 53 kD and the cbh2 consists of a fungal-type carbohydrate binding domain (CBD) separated from a catalytic domain by a linker region rich in proline/serine/threonine. PCR product consisting of the entire CBH II coding region without its signal sequences was cloned into the yeast secretive plasmid pPIC9K, which was then transformed into Pichia pastoris GS115. Highly efficient production of the cellobiohydrolase II was achieved in P. pastoris under the control of the AOX1 promoter, and the expressing level was 1.2 mg/mL by small-scale culturing. The recombinant cellobiohydrolase II was purified by using ammonium sulfate fraction, DEAE-Sepharose Fast flow chromatography. A molecular mass of the purified enzyme is 67 kD determined by SDS-PAGE and this is similar to the native cellobiohydrolase II purified from C. thermophilum CT2. The recombinant enzyme exhibited optimum catalytic activity at pH 4.0 and 50 degrees C respectively. It was thermostable at 50 degrees C and retained 50% of its original activity after 30 min at 70 d degrees C . The high level of fully active recombinant cellobiohydrolase II got from P. pastoris makes this expression system attractive for fermentor and industrial applications.
Amino Acid Sequence ; Base Sequence ; Cellulose 1,4-beta-Cellobiosidase ; biosynthesis ; genetics ; Chaetomium ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Fungal Proteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo investigate the effects of butyl alcohol extract of Baitouweng decoction ( BAEB) on yeast-to-hyphae transition of Candida albicans isolates from vulvovaginal candidiasis (VVC) in alkaline pH.

METHODSerial 2-fold dilution assay was used to determine the minimal inhibitory concentrations (MICs) of Baitouweng decoction extracts against C. albicans isolates from VVC, XTT assay was applied to determine the metabolic activity of C. albicans hypha treated by BAEB for 6 h. The morphological change of C. albicans treated by BAEB was inspected at different pH by inverted microscope, fluorescence microscope, scanning electron microscopy (SEM). Solid agar plate and semi-solid agar were utilized to evaluate colony morphology and invasive growth of C. albicans, respectively. Quantitative Real-time PCR (qRT-PCR) was adopted to observe the expressions of hyphae-specific genes including HWP1, ALS3, CSH1, SUN41 and CaPDE2.

RESULTThe MIC of BAEB against C. albicans is less than that of other extracts; hyphae grow best at pH 8. 0; 512 mg · L(-1) and 1,024 mg · L(-1) BAEB could inhibit formation of hyphae and influence colony morphology. When treated by 512 mg · L(-1) and 1,024 mg · L(-1) BAEB, the colonies became smooth; while by 0 and 256 mg · L(-1) BAEB, the colonies became wrinkled. In semi-solid agar, the length of hyphae decreased steadily as the concentration of BAEB lowered. The expression of HWP1, ALS3, CSHl, SUN41 were downregulated by 5.12, 4.26, 3.2 and 2.74 folds, and CaPDE2 was upregulated by 2.38 fold.

CONCLUSIONBAEB could inhibit yeast-to-hyphae transition of C. albicans isolates from VVC in alkaline pH.

Antifungal Agents ; isolation & purification ; pharmacology ; Candida albicans ; drug effects ; genetics ; growth & development ; Candidiasis, Vulvovaginal ; drug therapy ; microbiology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Humans ; Hydrogen-Ion Concentration ; Hyphae ; drug effects ; growth & development

OBJECTIVETo investigate the effects of retroviral vector containing shRNA targeting rat angiotensin II type 1 receptor (AT1R) gene (Ad5-AT1R-shRNA) on blood pressure and AT1R mRNA expression in spontaneously hypertensive rats (SHR).

METHODSRetroviral vector containing shRNA targeting rat AT1R gene was constructed and propagated further in 293 cells. SHR rats were randomly divided into SHR + Ad5-AT1R-shRNA (1.7 x 10(9) TCID(50)/ml) group and SHR (Ad5-EGFP, 7.9 x 10(9) TCID(50)/ml, n = 11 each) and 11 male Wistar-Kyoto rats (WKY) serve as normal controls (Ad5-EGFP, 7.9 x 10(9) TCID(50)/ml). Systolic blood pressure was measured before and after single intravenous injection of Ad5-AT1R-shRNA or Ad5-EGFP. Heart, liver, kidney, aorta and adrenal gland were removed after blood pressure measurement. Tissue Ad5-AT1R-shRNA expression was detected with fluorescence microscope and AT1R mRNA in liver, kidney and aorta was measured by fluorescence quantitative PCR.

RESULTSAd5-AT1R-shRNA significantly reduced blood pressure compared with controls (-29 mm Hg, 1 mm Hg = 0.133 kPa, P < 0.05) 24 hours after single injection and this antihypertensive effect could last for 5 to 7 days. Ad5-AT1R-shRNA expression detected with fluorescence microscope was significantly increased in heart, liver, kidney, aorta and adrenal gland post Ad5-AT1R-shRNA injection. AT1R mRNA in kidney and aorta (0.086 +/- 0.014, 0.051 +/- 0.023) were significantly decreased in Ad5-AT1R-shRNA treated rats compared with SHR control rats (0.362 +/- 0.042, 0.463 +/- 0.045, P < 0.01).

CONCLUSIONThe results indicate that Ad5-AT1R-shRNA could inhibit the tissue AT1R mRNA expression and produce prolonged antihypertensive effects in SHR rats.

Adenoviridae ; Animals ; Blood Pressure ; Genetic Vectors ; Heart Rate ; Hypertension ; genetics ; metabolism ; physiopathology ; Male ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism

In order to study the microstructure characteristics of normal lunate bones,eight fresh cadaver normal lunates were scanned with micro-computed tomography.High-resolution images of the micro-structure of normal lunates were obtained and we analyzed the nutrient foramina.Then nine regions of interest (ROI) were chosen in the central sagittal plane so that we could obtain the parameters of trabecular bones of ROIs.The distal lamellar-like compact structure had statistically significant differences when it was compared with the ROIs in the volar and dorsal ends of the distal cortex.The difference of diameter between the volar and dorsal foramina was significant (P＜0.05).However,there was no significant difference regarding the number.The trabecular bones of the volar and dorsal distal ends had lower intensity than those of the distal central subchondral bone plate.The diameters of the nutrient foramina on the volar cortex were larger than those on the dorsal.This research provided more detailed information about microstructure of normal lunate and the nutrient foramina on cortex,and a reference for further study about diseased lunate.

Objective To investigate the clinical significance of external obturation with cotton ball in treatment of digestive tract lip shape fistula. Methods A prospevtive randomixed controllde study was conducted in 30 patients with digestive tract lip shape fistula admitted from January 2001 to November 2007,which were divided into experimental group ( 15 patienets) and control griup ( 15 patjients). The control group received traditional fistula mouth continuous drainage,with fastomg or part enteral nutrition and parenteral nutrition. The experimernal group received external obturatiion with cotton ball,with controling spills of oineteatinal fluid and resuming normal eating and activetes, The patients general information, fistuala mouth sixe, volume, weight, albumin, determinstic operation time, postoperative complications, hospital, signficantly reduced fistula, flow, postoperative compared between the two groups. Results Compared with the control group , significantly redced fistula, flow, postoperative complivations and cost, increased wdighe and albumin,as well as shortened operation time and hospital stay were found oin the experimetal grou. Conclusin External obturation with cotton ball is safe effective methde in treatment of digestive tract lip shape fistula, obviusly shortening the course of disease, redcing patients'cost and pain, and rapidly improving patients nutritional status.