乾隆断案

乾隆断案孙雨亭，一也人物表（以出场先后为序）甘凤池──一等御前带刀侍卫。常升──南书房总管太监。乾隆──清高宗弘历。小贩──卖鸭血汤的。周青──乾隆后认的干儿子。张大宝──江宁府百姓，弱智青年。王升──王翠莲家仆人。王禄──王翠莲家仆人。徐氏──张大...

Wireless capsule video endoscopy:Three years of experience

AIM:To review and summerize the current literatue regarding M2A wireless capsule endoscopy. METHODS:Peer reviewed publications regarding the use of capsule endoscopy as well as our personal experience were reviewed. RESULTS:Review of the literature dearly showed that capsule endoscopy was superior to enteroscopy,small bowel follow through and computerized tomography in patients with obscure gastrointestinal bleeding,iron deficiency anemia, or suspected Crohn's disease.It was very sensitive for the diagnosis of small bowel tumors and for survailance of small bowel pathology in patients with Gardner syndrome or familial adenomatous polyposis syndrome.Its role in celiac disease and in patients with known Crohn's disease was currently being investigated. CONCLUSION:Capsule video endoscopy is a superior and more sensitive diagnostic tool than barium follow through, enteroscopy and entero-CT in establishing the diagnosis of many small bowel pathologies.

Diagnosis of intestinal tuberculosis using a monoclonal antibody to Mycobacterium tuberculosis

AIM:To investigate the utility of immunohistochemical(IHC) staining with an antibody to Mycobacterium tuberculosis(M.tuberculosis) for the diagnosis of intestinal tuberculosis(TB).METHODS:We retrospectively identified 10 patients(4 males and 6 females;mean age = 65.1 ± 13.6 years) with intestinal TB.Clinical characteristics,including age,gender,underlying disease,and symptoms were obtained.Chest radiograph and laboratory tests,including sputum Ziehl-Neelsen(ZN) staining,M.tuberculosis culture,and sputum polymerase chain reaction(PCR) for tubercle bacilli DNA,as well as Tuberculin skin test(TST) and QuantiFERON-TB gold test(QFT),were examined.Colonoscopic records recorded on the basis of Sato's classification were also reviewed,in addition to data from intestinal biopsies examined for histopathological findings,including hematoxylin and eosin staining,and ZN staining,as well as M.tuberculosis culture,and PCR for tubercle bacilli DNA.For the present study,archived formalin-fixed paraffin-embedded(FFPE) intestinal tissue samples were immunohistochemically stained using a commercially available species-specific monoclonal antibody to the 38-kDa antigen of the M.tuberculosis complex.These sections were also stained with the pan-macrophage marker CD68 antibody.RESULTS:From the clinical data,we found that no patients were immunocompromised,and that the main symptoms were diarrhea and weight loss.Three patients displayed active pulmonary TB,six patients(60%) had a positive TST,and 4 patients(40%) had a positive QFT.Colonoscopic findings revealed that all patients had type 1 findings(linear ulcers in a circumferential arrangement or linear ulcers arranged circumferentially with mucosa showing multiple nodules),all of which were located in the right hemicolon and/or terminal ileum.Seven patients(70%) had concomitant healed lesions in the ileocecal area.No acid-fast bacilli were detected with ZN staining of the intestinal tissue samples,and both M.tuberculosis culture and PCR for tubercle bacilli DNA were negative in all samples.The histopathological data revealed that tuberculous granulomas were present in 4 cases(40%).IHC staining in archived FFPE samples with anti-M.tuberculosis monoclonal antibody revealed positive findings in 4 patients(40%);the same patients in which granulomas were detected by hematoxylin and eosin staining.M.tuberculosis antigens were found to be mostly intracellular,granular in pattern,and primarily located in the CD68 + macrophages of the granulomas.CONCLUSION:IHC staining with a monoclonal antibody to M.tuberculosis may be an efficient and simple diagnostic tool in addition to classic examination methods for the diagnosis of intestinal TB.

^(99)Tc^m direct labeling of angiostatin

Objective: To explore the method of 99 Tc m direct labeling of angiostatin (AS) and investigate the stability and bioactivity of the 99 Tc m labeled AS in vitro . Methods: AS was extracted, validated, and then labeled with 99 Tc m after having been reduced by 2 ME or SnCl 2. The best labeling condition was screened by cross design. The labeling efficiency was measured by TLC and column chromatography. The stability of 99 Tc m AS was observed and compared when BSA, saline and different molar ratios of Cys∶AS were separately added. The bioactivity of 99 Tc m AS was observed in human umbilical vein endothelial cell (CEV304). Results: The labeling efficiency can reach (97±1 5)% for the 2 ME reducing approach. Its best experimental condition was as follows: AS 100 μg,PB(0 5 mol/L, pH 7 3)1 ml, 2 ME 100 μg, MDP (dissolved in 1 ml saline) 10 μl, and 99 Tc mO 4 - 185 MBq. The labeling efficiency using SnCl 2 reducing method can reach (90±3 0)%. The best experimental procedure was as follows: AS 100 μg,boric acid buffer(0 1 mol/L, pH 9 0)1 ml, 2%SnCl 2 (dissolved in 1 mol/L hydrochloric acid) 20 μl, was added into MDP, which was diluted with 1 ml deoxygenized water, and then 20 μl, 99 Tc mO 4 - 185 MBq was added. The product of 99 Tc m labeled AS was stable in vitro and had the same bioactivity as AS. Conclusion: 99 Tc m direct labeling of AS is simple and efficient. And the bioactivity of 99 Tc m AS has no significant change compared with AS.

Serial Expression of the Truncated Fragments of the Nucleocapsid Protein of CCHFV and Identification of the Epitope Region

The Crimean-congo hemorrhagic fever virus(CCHFV)is a geographically widespread fatal pathogen. Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections.In this study,expression vectors carrying series truncated fragments of the NP(nucleocapsid protein)gene from the S fragment of CCHFV strain YL04057 were constructed.The recombinant proteins were expressed in E.coli and purified for detection.The antigenic of the truncated fragments of NP was detected with a polyclonal serum(rabbit)and 2 monoclonal(mAbs)(14B7 and 43E5)against CCHFV by Western-blot analyses. The results showed that the three expressed constructs,which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum.The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein.

Characteristics of Fault Activity in the Taiyangshan Uplift Area in Hunan,China

Lying on the west edge of Dongting Lake,the Taiyangshan area in Hunan Province is part of a hilly region which has uplifted since the Late Cenozoic.According to field investigation of the six existing faults in the Taiyangshan area,we found that four of them are not active in the Quaternary,and that the Gangshi-Hefu fault is likely to have been active in the early Mid-Pleistocene.The geological evidence derived suggests that the Xiaowupu fault was active from the late Mid-Pleistocene to the early late-Pleistocene.It cut the stratum with a TL age of 123±10ka BP and has the property of thrusting.The research results are of great significance for understanding the seismogenic structure of the Changde earthquake with M6 3/4 in 1631.

Cloning of NHE-1 gene fragment from human lung cancer cells and construction of its antisense expression vector

Objective: To clone the partial sequence of N+/H+ exchanger-1 (NHE-1) gene of human lung cancer cells and insert it reversely into the multiclone site of pLXSN in order to construct an antisense expression vector for tumor gene therapy in vivo. Methods: With use of the upstream and downstream primers containing Barn H I and EcoR I in their5’ ends respectively, a partial sequence of the first exon of NHE-1 gene was cloned in a length of 454 bp from genomic DNA of human lung cancer cell A549 with PCR method. The product was then directionally and reversely insert into the multiclone site of pLXSN. Finally, the constructed recombinant was identified with agarose gel electrophoresis and DNA sequencing. Results: The cloned fragment was 461 bp in length and successfully ligated to pLXSN with the identification by agarose gel electrophoresis. DNA sequencing confirmed that the fragment cloned and inserted into the vector was identical with the targeted one. Conclusion: The targeted fragment is successfully cloned and reversely inserted into pLXSN in our experiment. The antisense expression vector of NHE-1, pNHE-1. was consfructed successfully.

Predominance of Plasmodium malariae-falciparum Co-Infection by Molecular Speciation in Bangolan, North West Region of Cameroon

Although microscopy still remains the gold standard for the diagnosis of malaria, rapid diagnostic tests （RDTs） and PCR assays have been shown to be sensitive and specific. Very few comparative studies have been reported of the three diagnostic methods on the same samples in vulnerable groups. Microscopy, RDTs and PCR assays were used for detection and speciation of Plasmodium falciparum （P）＇）, Plasmodium malariae （Pm） and Plasmodium ovale （Po） in patients in a rice culture savanna ecotype. Fifty four children and 16 pregnant women presenting with a fever were recruited. Bloods collected was used for thin and thick smears, perform RDTs and spotted blood on filter paper for DNA extraction and performance of a PCR. Mean parasitaemia was 37,619.06 （＋ 33,599.04） p/pL and 7,512.5 （＋ 12,446.11） p/μL for children and pregnant women, respectively. A total of 87.14% were positive by microscopy, 85.71% by RDTs and 90% by PCR. Distribution of Plasmodium species as identified by PCR was 72.86% Pf/Pm, 11.43% Pf/Pm/Po and 5.43% Pm while 10% were negative. Cohen＇s Kappa value for PCR and RDTs was K = 0.75 （CI = 0.28-1.22） while PCR and microscopy was K = 0.64 （CI = 0.18-1.10）. Malaria infection in Bangolan was mostly due to mix infection predominantly P. falciparum/P, malariae.

Terraces and Tectonic Uplifts at Yandantu and Changcaogou on the Northern Margin Fault of Altun

The formation of strath and strath terrace is closely related to tectonic uplift in the drainage basin. Based on the investigation of straths at Yandantu and Changcaogou on the eastern segment of the northern margin fault of Altun, and in combination with the paleoclimatic data, the tectonic uplift since late Epipleistocene as revealed by stream terraces at the two places is discussed. At Yandantu, three levels of stream terraces(T 1, T 2 and T 3)have developed since 16ka BP, where T 1, T 3 and T 2 are fill terraces and the buried major straths are exposed. The ages of three treads are dated to be about 16.1ka BP, 12.8ka BP and 6.2ka BP, respectively. The three terraces reflect three tectonic uplift events, while the ages of the treads represent the occurrence time of these events. The stream is still beveling the bedrock and widening the channel at present, and the modern strath is being generated. The uplift rate is 4.8～4.5mm/a since 16.1 ka BP in this area. From 12.8ka B.P to 6.2ka BP, The uplift rate was 6.4mm/a. The uplift rate is 3.1mm/a since 6.2ka BP. At Changcaogou, four levels of stream terraces(T 1, T 2, T 3 and T 1′)have developed since 7ka BP. All of them are fill terraces. There are buried straths under the deposits. The buried major strath is exposed on T 3 and T 2 and the minor strath on T 1′and T 1. The ages of treads of the three terraces (T 3, T 2 and T 1′) are 7 ka BP, 3 ka BP and 2.5 ka BP, respectively. The four terraces reflect two uplift events induced by tectonic activities. One occurred in about 7 ka BP, and the other in 3ka BP. The uplift rate is 5.9mm/a since 7.0 ka BP at Changcaogou. From 7ka BP to 3ka BP, the uplift rate was 7.0mm/a, and since 3ka BP till now, the uplift rate is 4.7 mm/a.

Development of a Polyclonal Antibody-based AC-ELISA and Its Comparison with PCR for Diagnosis of Canine Parvovirus Infection

A polyclonal antibody-based antigen-capture ELISA （AC-ELISA） has been developed for detection of Canine parvovirus （CPV） antigens in faecal samples of dogs. The assay uses rabbit anti-CPV polyclonal antibody as the capture antibody, guinea pig anti-CPV polyclonal antibody as tracing antibody and anti-guinea pig HRPO conjugate as the detection system. The optimum dilution of the capture antibody and the tracing antibody capable of detecting the CPV-2 antigens was found to be 1：1 600 and 1：400, respectively, in the check-board titration. In this study, a total of 152 samples （129 faecal samples and 23 cell culture supernatant） were tested both by AC-ELISA and by polymerase chain reaction （PCR）. Of the samples tested, 69 and 78 samples were found positive by AC-ELISA and PCR, respectively. The AC-ELISA had relative sensitivity, relative specificity and accuracy of 88.4%, 100.0% and 91.4% respectively. The analytical sensitivity of AC-ELISA was estimated to be 102.8 TCID50/mL whereas PCR sensitivity was 100.8 TCIDs0/mL. The AC-ELISA is a simple, quick and reliable method for screening large numbers of faecal samples of dogs suspected of CPV infection.

Cloning and characterization of a novel gene encoding a putative seven-span transmembrane protein localized in endoplasmic reticulum

Objective: To clone and analyze the structure of a novel gene, named EST 1 (endoplasmic reticulum localized seven span transmembrane protein 1) and to analyze the expression pattern and intracellular location of EST 1. Methods: The cDNA library was screened to isolate novel cDNA fragment. The structure of novel gene was analysed by computer software. Expression of EST 1 was analyzed by dot blot and Northern blotting. Intracellular localization was observed after EST 1 enhanced green fluorescence protein (EGFP) fusion gene was transfected into mammalian cells. Results: The full length cDNA of mouse EST 1 was 1 802 bp, with a 1 293 bp open reading frame encoding 431 amino acids. It was predicated that protein encoded by EST 1 contained a signal peptide sequence at the N terminus, seven putative transmembrane domains, and an ER retaining signal at the C terminus. EST 1 EGFP fusion protein showed an ER like intracellular distribution in mammalian cells. Expression pattern analysis showed that EST 1 is expressed in all tissues examined. Conclusion: EST 1 is encoding a putative seven span transmembrane protein localized in endoplasmic reticulum. EST 1 was expressed in all tissues examined, suggesting an essential function of EST 1 in cells.

Establishment of a Two-site ELISA for Detection of Chlamydia Trachomatis

Objective:To establish a rapid and simple assay for the diagnosis of Chlamydia trachomatis (CT) infection. Methods BALB/c mice were immunized with SDS-PAGE purified major outer membrane protein (MOMP) from CT and the monoclonal antibodies were obtained subsequently. Two-site ELISA was developed to detect CT infection. Results: The established assay was able to detect as low as 1.248ug/ml MOMP with interrun and inrun CV 6.9% and 3.1% respectively. 94% (34/36) of culture-positive samples were found to be positive in the current examination, indicating the high sensitivity of this assay. Conclusion: The assay is applicable for clinical diagnosis of CT infection.

ISO9002走进旅游酒店业

中国旅游酒店业自改革开放以来,经历了从无到有、从追求数量到追求质量、从卖方市场到买方市场转变的过程。短短二十年,走过了西方国家几十年甚至上百年的历程。据权威部门统计,到1997年春,我国的旅游酒店已增加到近5000家,共有65万多个房间,平均每年的增长幅度高达18.96％。二十年间,我国旅游酒店不仅在数量上翻了五番之多,而且管理水平、服务质量也大为提高,初步实现了与国际惯例相接轨。

A stress-rifting origin of Grand Canyon

The Grand Canyon is a massive rift in the Colorado Plateau. How and when it developed has been debated for nearly 150 years. Most geologists believe the unusual landscape was primarily shaped by water erosion.Here we propose a stress-rifting model to provide an alternative explanation for the origin of Grand Canyon.This paper adopts a brittle–ductile double layer model to simulate the deformation and rifting of the plateau due to the mantle-melting-induced expansion. Our results show that the uplift induced by thermal expansion and its associated horizontal extension can cause open fractures that extend from the brittle surface to the underlying ductile layer in a top-down way. In addition, we find that episodic uplift can deepen and connect multiple fractures together to form a larger fracture network. Our findings suggest that the formation of the Grand Canyon might have been driven by plateau uplift and its associated rifting under crustal extension, wherein water erosion played only a minor role in shaping the course of the Colorado River. The new paradigm provides simpler explanations to some of the long-standing geological mysteries surrounding the canyon.