In this study, we developed an efficient electroporation-mediated transformation system featuring Flammulina velutipes. The flammutoxin (ftx) gene of F. velutipes was isolated by reverse transcription-PCR. pFTXHg plasmid was constructed using the partial ftx gene (410 bp) along with the hygromycin B phosphotransferase gene (hygB) downstream of the glyceraldehydes-3-phosphate dehydrogenase (gpd) promoter. The plasmid was transformed into protoplasts of monokaryotic strain 4019-20 of F. velutipes by electroporation. High transformation efficiency was obtained with an electric-pulse of 1.25 kV/cm by using 177 transformants/microg of DNA in 1 x 107 protoplasts. PCR and Southern blot hybridization indicated that a single copy of the plasmid DNA was inserted at different locations in the F. velutipes genome by non-homologous recombination. Therefore, this transformation system could be used as a useful tool for gene function analysis of F. velutipes.
Agaricales ; Blotting, Southern ; Chimera ; Cinnamates ; Coat Protein Complex I ; DNA ; Electroporation ; Flammulina ; Fungal Proteins ; Genome ; Hygromycin B ; Mycotoxins ; Oxidoreductases ; Plasmids ; Polymerase Chain Reaction ; Protoplasts ; Recombination, Genetic ; Sprains and Strains

PURPOSE: To examine the effects of change in the weight bearing on the growth plate metabolism, a simulated animal model of weightlessness was introduced and the chondrocytes' cellular kinetics were evaluated. MATERIALS AND METHODS: Unloading condition on the hind-limb of Sprague-Dawley rats was created by fixing a tail and lifting the hind-limb. Six rats aged 6 weeks old were assigned to each group of unloading, reloading, and control groups of unloading or reloading. Unloading was maintained for three weeks, and then reloading was applied for another one week thereafter. Histomorphometry for the assessment of vertical length of the growth plate, 5-bromo-2'-deoxyuridin (BrdU) immunohistochemistry for cellular kinetics, and biotin nick end labeling TUNEL assay for chondrocytes in the growth plate were performed. RESULTS: The vertical length of the growth plate and the proliferative potential of chondrocytes were decreased in the unloading group than those of control groups. Inter-group differences were more significant in the proliferative and hypertrophic zones. Reloading increased the length of growth plate and proliferative potential of chondrocytes as evidenced by increase of the ratio of positive BrdU stained cells. However, apoptotic changes in the growth plate were not affected by the alterations of weight bearing. CONCLUSION: Alterations in the weight bearing induced changes in the chondrocytic proliferative potential of the growth plate and have no effects on the apoptosis occurred. This may suggest that deprived weight bearing due to various clinical situations hamper normal longitudinal bone growth, and further studies regarding the factors for reversibility of chontrocytic proliferation upon variable mechanical stresses are needed.
Animals ; Apoptosis ; Biotin ; Bone Development ; Bromodeoxyuridine ; Chondrocytes ; Growth Plate* ; Immunohistochemistry ; In Situ Nick-End Labeling ; Kinetics* ; Lifting ; Metabolism ; Models, Animal ; Rats ; Rats, Sprague-Dawley ; Stress, Mechanical ; Tail ; Weight-Bearing* ; Weightlessness

BACKGROUND: To examine the effects of changes in mechanical loading on endochondral bone formation, a simulated rat model of weightlessness was introduced and the changes in the growth plate were evaluated. METHODS: Unloading condition on the hindlimb of Sprague-Dawley rats was created by fixing a tail and lifting the hindlimb. Six rats aged 6 weeks old were assigned to each group of unloading and reloading with their control group. Unloading was maintained for three weeks, and then reloading was applied for another one week afterwards. Histomorphometry for the assessment of vertical length of the growth plate, 5-bromo-2'-deoxyuridin (BrdU) immunohistochemistry for cellular kinetics, and biotin nick end labeling TUNEL assay for chondrocytes in the growth plate were performed. RESULTS: The vertical length of the growth plate and the proliferative potential of chondrocytes were decreased in the unloading group than those of the control group. Inter-group differences were more significant in the proliferative and hypertrophic zones. Reloading increased the length of growth plate and proliferative potential of chondrocytes as evidenced by the increase of the ratio of positive BrdU stained cells. However, the apoptotic changes in the growth plate were not affected by the alterations of the weight bearing. CONCLUSION: Alterations in the weight bearing induced changes in the chondrocytic proliferative potential of the growth plates and had no effect on the apoptosis occurrence. This may suggest that deprived weight bearing due to various clinical situations hamper normal longitudinal bone growth. Further studies regarding the factors for reversibility of chondrocytic proliferation upon variable mechanical stresses are needed.
Animals ; Apoptosis ; Biotin ; Bone Development ; Bromodeoxyuridine ; Chondrocytes ; Growth Plate ; Hindlimb ; Immunohistochemistry ; In Situ Nick-End Labeling ; Kinetics ; Lifting ; Models, Animal ; Osteogenesis* ; Rats* ; Rats, Sprague-Dawley ; Stress, Mechanical ; Tail ; Weight-Bearing ; Weightlessness

OBJECTIVE: To provide clinical information for the best diagnosis and treatment of primary malignant cervical lymphoma based on the information obtained from these cases. METHODS: Between 1989 and 2006, six women with primary malignant cervical lymphoma were diagnosed and treated at our institution. Data were obtained from their medical records and were retrospectively analyzed. RESULTS: The mean patient age at the time of diagnosis was 63 (range 19-74). The chief complaint was vaginal bleeding in five women and the voiding difficulty for one woman. five of six patient had the cervical lesions (erosion and tumoral mass), while the other was non specific cervical findings. The Papanicolaou test was performed on three women, one of whom was HSIL. All six patients were confirmed with cervical lymphoma through the pathologic diagnosis. Surgical treatment (radical hysterectomy with bilateral salpingo- oophorectomy, pelvic lymph node dissection, and para-aortic lymph node dissection) was performed in one case, simple hysterectomy with complementary chemotherapy in two, and chemotherapy in one. Two patients refused treatment. Among four patients treated, one experienced recurrence in an ovary and the others have remained in remission. CONCLUSION: Primary malignant cervical lymphoma is a rare malignancy. Physicians can miss early detection of this disease because of its 'silent' symptoms and very low incidence. The accumulated data regarding this tumor can make it easy to detect at an early stage, thereby allowing it to be successfully treated. Further studies should be conducted to obtain further information regarding the cervical lymphoma.
Cervix Uteri ; Diagnosis ; Drug Therapy ; Female ; Humans ; Hysterectomy ; Incidence ; Lymph Node Excision ; Lymph Nodes ; Lymphoma* ; Medical Records ; Ovariectomy ; Ovary ; Papanicolaou Test ; Recurrence ; Retrospective Studies ; Uterine Hemorrhage

PURPOSE: X-ray film over responds to low-energy photons in relative photon beam dosimetry because its sensor is based on silver bromide crystals, which are high-Z molecules. This over-response becomes a significant problem in clinical photon beam dosimetry particularly in regions outside the penumbra. In intensity modulated radiation therapy (IMRT), the radiation field is characterized by multiple small fields and their outside-penumbra regions. Therefore, in order to use film dosimetry for IMRT, the nature the source of the over-response in its radiation field need to be known. This study is aimed to verify and possibly improve film dosimetry for IMRT. MATERIALS AND METHODS: Modulated beams were constructed by a combination of five or seven different static radiation fields using 6 MeV X-rays. In order to verify film dosimetry, we used X-ray film and an ion chamber were used to measure the dose profiles at various depths in a phantom. In addition, in order to reduce the over-response, 0.01 inch thick lead filters were placed on both sides of the film. RESULTS: The measured dose profiles showed a film over-response at the outside-penumbra and low dose regions. The error increased with depths and approached 15% at a maximum for the field size of 15X15 cm(2) at 10 cm depth. The use of filters reduced the error to 3%, but caused an under-response of the dose in a perpendicular set-up. CONCLUSIONS: This study demonstrated that film dosimetry for IMRT involves sources of error due to its over-response to low-energy photons. The use of filers can enhance the accuracy in film dosimetry for IMRT. In this regard, the use of optimal filter conditions is recommended.
Film Dosimetry* ; Photons ; Silver ; X-Ray Film

Malignant mixed mullerian tumor (MMMT) is rare and has pathologically carcinoma and sarcoma components. Among them, primary peritoneal MMMTs are extremely rare, and their proper treatment and prognosis are not well-known. We experienced a case of primary peritoneal origin MMMT, so we report it with a brief review of the literatures.
Peritoneum ; Prognosis ; Sarcoma

The goal of the immunologic maneuvering for organ transplantation may be the donor specific immune tolerance rather than non-specific immunosuppression. Although DST is one of the most extensively studied methods for donor specific immune hyporesponsiveness, it is not widely used in recent years because of possible sensitization and overall improvement of graft survival without DST. Several human and animal studies showed that -24h DST with concomitant cyclosporine administration improved graft survival. -24 DST may not induce adverse sensitization that preclude subsequent transplantation and the procedure is simple and does not delay the operation in living donor transplantation. Between Feb. 1994 and Jan. 1997, 33 patients received 100-200ml of fresh whole blood from the kidney donor 1 day before transplantation. Twenty donors were living related and 13 donors were non- related. Mean age was 40(22-52). Two patients was diabetic. All but one received primary allograft. Cyclosporine and prednisolone were the primary immunosuppressants that started 2-3 days before transplantation. Acute rejection occurred in 11 recipients(33.3%). Acute rejection tended to occur earlier. Eight of 11 first episodes were within 3 days post- transplant, which were all recovered by either steroid pulse or OKT3. Mean follow up was 35 months. Two patients died with functioning graft. Three-year graft survival rate was 93.9%. There was no immunologic graft loss. We conclude that -24h DST may be a valuable option of immune modulation for renal transplantation with no demonstrable adverse reaction. It's beneficial effect needs to be confirmed by a larger controlled study.
Allografts ; Animals ; Cyclosporine ; Follow-Up Studies ; Graft Survival ; Humans ; Immune Tolerance ; Immunosuppression ; Immunosuppressive Agents ; Kidney ; Kidney Transplantation* ; Living Donors* ; Muromonab-CD3 ; Organ Transplantation ; Prednisolone ; Tissue Donors* ; Transplants