This paper summarize the practice in the teaching reform of microbiology experiment in recent years. We identify the main contents of experimental teaching systems and pay much more attention to peo-ple-oriented. Through the reform of teaching and assessment methods,students are trained to cultivate their practical ability and spirit of innovation.

In this study, plate transparent circle by Davis method was introduce firstly screening ?-Rhamnosidase high-yield strain. The spore-sprouted Aspergillus niger 8-hour were mutagenized by ethyl methane sulphonate and pre-screened via transparent circle. 11% mutants yield 40% higher of ?-rhamnosidase than the original strain. A high-yield strain, T-226 with the highest ?-rhamnosidase activity of 373.4 U/mL was finally selected from these potential high-yield mutants after rescreened by shake flask fermentation twice. When the T-226 strain was fermented in 5 L bioreactor, the enzyme activity could reach to 631.9 U/mL after 84 h. Thus, the established screening method is highly efficient to isolate ?-rhamnosidase high-yield mutant of A. niger.

OBJECTIVETo emphasis the importance of recognizing and treating the early colorectal carcinoma with the invasion limited in the layer of muscularis mucosae, and discuss several questions in using the WHO's new diagnostic standards of the prestage of tumors of digestive system.

METHODSA retrospective review was made of 30 patients with invasive carcinoma limited in the layer of muscularis mucosae. All of the patients were treated between July 1986 and July 1999 in Shanghai Cancer Hospital.

RESULTSAmong the 30 cases, there were 19 men and 11 women from 20 to 80 years dd (mean, 57.0). The tumor diameter was ranged from 0.8 cm to 8.0 cm (mean, 2.9 cm). 12 patients were given polypectomy while 18 patients were given radical resection. Among the patients given polypectomy, one got recurrence 3 years later, one had the metastasis of lungs 50 months later and had been dead already. Among the patients given radical resection, one had metastasis of lymph nodes peripheral to the tumor, one also had the metastasis of lungs 68 months later but is still alive now, and one had metastasis of lymph nodes in the right upper part of neck 15 months later and then was lost to follow-up.

CONCLUSIONSThe patients with early colorectal carcinoma with the invasion limited in the layer of muscularis mucosae could have metastasis of lymph nodes, get local recurrence after polypectomy and even have blood metastasis. Therefore great attention must be paid. Clinicians should be careful to chosen polypectomy. Meanwhile, in order to provide more information of the correct treatment, it will be better if the pathologists could give the original diagnosis at the same time when they use the WHO's new diagnostic standards of the prestage of tumors of digestive system in diagnosing the early invasive carcinoma limited in the layer of muscularis mucosae, for the concept of high-grade intraepithelial neoplasia will be used instead of it, and whether or not it is a really benign tumor, much more clinical research must be done later.

Adult ; Aged ; Aged, 80 and over ; Colectomy ; methods ; Colorectal Neoplasms ; pathology ; surgery ; Female ; Humans ; Intestinal Mucosa ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Rectum ; surgery ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo emphasize the importance of correct and standardized surgical treatment on dermatofibrosarcoma protuberans (DFSP), and discuss the suitable synthesized therapy on it.

METHODS163 cases of DFSP, which were treated between January 1985 and September 2002,were submitted to a retrospective study.

RESULTSAmong the 163 cases, 150 (92.0%) were treated with local excision as benign tumors before accepted to Cancer Hospital, Fudan University. 69 cases (46.0%) were approved by pathological examination to have tumor remnants after they were treated with wide excision, and 49 (71.0%) of them couldn't be found to have any tumor remnants by physical examination or B-ultrasonic examination before that operation. It was easy for the tumor to recur after excision, especially the local excision. The recurrent rate after it was 45.1%, which was much higher than the one after wide excision (5.6%). Among the 142 cases which wide excision were performed, 99 ones had excision margins >/= 3 cm and 5 of them (5.1%) developed local recurrence while 36 ones had excision margins 1 approximately 2 cm and 3 of them (8.3%) developed local failure. 46 cases (32.4%) were given skin graft, 11 cases were given flap, and 1 case had dacron mending in skin defection area. The main complications after these operations were necrosis of the skin flap (20 cases) and infection of the wound (6 cases). They could all be cured in 2 months. 17 cases were given complimentary radiotherapy with the dose range from 3275 cGy to 7000 cGy because of their recurrences for times or positive resection margins after wide excision. Only one case had wet molting after radiotherapy and 2 developed local recurrence. Among all the 163 cases, only 2 (1.2%) were dead, and 1 of them was died of metastasis of lung and liver. 2 cases got lymph node metastasis, then were given surgical treatment and still alive now. 13 cases (8.0%) were DFSP-FS with their malignancies increased. 11 of them were the recurrent ones after local excision or wide excision.

CONCLUSIONSIn order to avoid misdiagnosis, it is necessary for the clinician to know much about DFSP. Once the tumor was diagnosed of DFSP after local excision, it is necessary to take wide excision. Because DFSP is a malignancy of a high recurrent rate after local excision, standardized wide excision is the key in reducing local failure. Adjuvant radiotherapy is an effective treatment for the patients with positive resection margin or the patients don't suit for surgical treatment. The DFSP-FS need to use more energetic treatment in curing it.

Adolescent ; Adult ; Aged ; Child ; Combined Modality Therapy ; Dermatofibrosarcoma ; diagnosis ; radiotherapy ; surgery ; therapy ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Skin Neoplasms ; diagnosis ; radiotherapy ; surgery ; therapy

In order to develop a rapid method which can check Campylobacter jejuni in animal and poultry foods nicely, an immunomagnetic capture-fluorescent PCR (IMC-FPCR) method was established in this paper. The reported method involves isolation of the target pathogen by immunocapture prior to the fluorescent PCR step, therefore the immunomagnetic-beads for Campylobacter were developed, and two groups of primer/probe, which targeted for the species special sequence of flaA gene and hipO gene for Campylobacter jejuni were designed. The immunomagnetic capture-fluorescent PCR assay amplification of the hipO gene and flaA gene for detection of Campylobacter jejuni was firstly reported in this paper. Result indicated that IMC-FPCR method permits direct detection of the pathogen without an enrichment step and can be performed in approximately 24 h. The assay results are positive for all of the isolates of Campylobacter jejuni (3 isolates, including type strain ATCC 33560 and ATCC8341) with a detection limit of approximately 10 cfu/mL, are negative for Campylobacter coli and several other bacteria. IMC-FPCR assay provide not only a rapid, sensitive method for quantitative detection of Campylobacter jejuni, but also an important method for detecting of Campylobacter jejuni of viable but non-culturerable (VNC) state.
Campylobacter jejuni ; genetics ; isolation & purification ; Fluorescence ; Immunomagnetic Separation ; methods ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

AIMTo explore the effects of tRNA on the growth of mammalian cells.

METHODSL929, NIH3T3, MCF-7 and PC12 cells were seeded in 96 well culture plate individually, and incubated at 37 degrees C in 5% CO2 for 4 h, the tRNAs from different species were added to the culture media individually. After certain time of incubation, the viability of the cells was evaluated by the MTT methods. Sub-confluent L929 cells were incubated with 200 microg/ml ytRNA for different times, then the cells were pooled and analyzed with flow cytometry assay.

RESULTStRNA specifically inhibited the growth of L929 cells in a dose-dependent manner. The sizes of tRNA-treated cells showed larger sizes and longer processes than those of untreated cells. Flow cytometric analysis further showed that most of tRNA-treated cells were arrested in S phase of the cell cycle.

CONCLUSIONThe cell growth inhibitory effects of tRNAs were caused mainly by their degraded fragments. The results suggested that tRNA or its degraded fragments might play important roles in regulation of cell proliferation.

Animals ; Cell Cycle Checkpoints ; physiology ; Cell Line ; Cell Proliferation ; Fibroblasts ; cytology ; Flow Cytometry ; Mice ; RNA, Transfer ; physiology

OBJECTIVETo study the incidence rate of multiple primary colorectal carcinomas (MPCC) in colorectal carcinoma and to evaluate its clinical and pathological characteristics.

METHODSOne hundred and sixty-eight (4.6%) patients from 3663 cases with colorectal carcinoma were diagnosed with MPCC from January 1985 to December 2003. The clinical data of the patients were collected retrospectively to investigate the diagnosis and treatment of MPCC.

RESULTSOf the 168 patients, 81 were diagnosed as synchronous colorectal carcinoma (SC), 72 with metachronous colorectal carcinoma (MC), 15 with both SC and MC. The median age at time of diagnosis of colorectal carcinoma was 58 years old (range from 20 to 82 years old). Three hundred and ninety-three cancer lesions were detected in these 168 cases (mean, 2.3 lesions/case). The rectum and sigmoid colon were the most involved sites (61.6%). Eighteen cases (10.7%) were verified with hereditary non-polyposis colorectal cancer (HNPCC) while another 9 cases were highly suspected. Fourteen patients (8.3%) were found with other malignancies out of large intestine, 41 patients (24.4%) with colorectal adenomas, 72 (42.9%) with adenoma carcinogenesis. Among the 96 SC patients, 91 were given preoperative colonoscopy and 65 (71.4%) got the diagnosis. All the MC patients were diagnosed by postoperative colonoscopy. The overall 5-year survival rate of the 168 patients was 69.8%.

CONCLUSIONSMPCC should be paid more attention in colorectal cancer management. Colonoscopic surveillance is much more important in diagnosis and follow-up of MPCC for reducing the misdiagnosis of SC and detecting more MC in time. Prompt treatment of adenoma can reduce the occurrence of MPCC, and active and standard surgical treatment should be done for MPCC.

Adult ; Aged ; Aged, 80 and over ; Colonoscopy ; Colorectal Neoplasms ; diagnosis ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasms, Multiple Primary ; diagnosis ; pathology ; surgery ; Retrospective Studies

TSO18 gene was subcloned into the Pichia pastoris expression vector pPIC9K. The recombinant plasmid pPIC9K-TSO18 was transformed into P. pastoris GS115 by electroporation so that the plasmid will be integrated with chromosome of P. pastoris. The P. pastoris strains containing multi-copy recombinant were screened by G418 and induced by methanol. The expression product was analyzed by SDS-PAGE, Western blot, deglycosylation, and purified by Sephadex column, and was used to immunize mice. The results indicated that the target protein was efficiently expressed in P. pastoris, and glycosylated moderately, and had immunological activity. In a 5 liter fermentor, the expression level of the target protein was up to 2.54 mg/mL. These results will benefit for the development of genetically engineering vaccine.
Animals ; Antigens, Helminth ; biosynthesis ; genetics ; immunology ; Cloning, Molecular ; Electroporation ; Gene Expression ; Genetic Vectors ; genetics ; Mice ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Swine ; Taenia solium ; genetics ; immunology

OBJECTIVETo summarize the clinical features of invasive pulmonary fungal infection (IPFI) secondary to malignant blood diseases (MBD).

METHODSWe retrospectively analyzed the clinical data of 52 patients with IPFI secondary to MBD admitted to Peking Union Medical College Hospital from January 1995 to December 2008.

RESULTSThe incidences of IPFI secondary to acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), non-Hodgkin's lymphoma (NHL), and aplastic anemia (AA) were 4.6%, 3.2%, 2.8%, and 2.5%, respectively. In patients with IPFI secondary to AML, 88.5% (23/26) of the patients suffered from the infections during the non-remission (NR) period (including relapse), and 11.5% (3/26) in the complete-remission (CR) period. In all the patients with IPFI secondary to malignant blood diseases, 86.5% (45/52) of MBD were neutropenic or agranulocytic, and 67.3% (35/52) had been treated with broad-spectrum antibiotics for more than 96 hours before anti-fungal therapy. The total mortality after anti-fungal therapy was 13.7% (7/51). More than half of patients with fluconazole or itraconazole as the first-line therapy had to switch to other medicines because of poor infection control.

CONCLUSIONSIPFI secondary to MBD is most common in AML patients. Patients with NR of AML, neutropenia or agranulocytosis, and long-term broad-spectrum antibiotics usage are susceptible to IPFI. Fluconazole and itraconazole have low efficacy, and other more potent anti-fungal medicines should be considered.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Hematologic Neoplasms ; complications ; Humans ; Lung Diseases, Fungal ; diagnosis ; drug therapy ; etiology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

BACKGROUNDAtopic dermatitis (AD) is an inflammatory skin disease characterized by chronic recurrent dermatitis with profound itching. Most patients have personal and/or family history of atopic diseases. Several criteria have been proposed for the diagnosis of AD. Although the clinical features of childhood AD have been widely studied, there has been less large-scale study on adult/adolescent AD. The aim of this study was to investigate the clinical features of adult/adolescent patients with chronic symmetrical eczema/AD and to propose Chinese diagnostic criteria for adult/adolescent AD.

METHODSA hospital-based study was performed. Forty-two dermatological centers participated in this study. Adult and adolescent patients (12 years and over) with chronic symmetrical eczema or AD were included in this study. Questionnaires were completed by both patients and investigators. The valid questionnaires were analyzed using EpiData 3.1 and SPSS 17.0 software.

RESULTSA total of 2662 valid questionnaires were collected (1369 male and 1293 female). Of all 2662 patients, 2062 (77.5%) patients had the disease after 12 years old, while only 600 (22.5%) patients had the disease before 12 years old, suggesting late-onset eczema/AD is common. Two thousand one hundred and thirty-nine (80.4%) patients had the disease for more than 6 months. One thousand one hundred and forty-four (43.0%) patients had a personal and/or family history of atopic diseases. One thousand five hundred and forty-eight (58.2%) patients had an elevated total serum IgE and/or eosinophilia and/or positive allergen-specific IgE. Based on these clinical and laboratory features, we proposed Chinese criteria for adult/adolescent AD. Of all 2662 patients, 60.3% were satisfied with our criteria, while only 48.2% satisfied with Hanifin Rajka criteria and 32.7% satisfied with Williams criteria, suggesting a good sensitivity of our criteria in adult/adolescent AD patients.

CONCLUSIONLate-onset of eczema or AD is common. The clinical manifestations of AD are heterogeneous. We have proposed Chinese diagnostic criteria for adolescent and adult AD, which are simple and sensitive for diagnosis of adult/adolescent AD.

Adolescent ; Adult ; Dermatitis, Atopic ; diagnosis ; immunology ; Eczema ; diagnosis ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Middle Aged ; Retrospective Studies ; Surveys and Questionnaires