[Detection of MDM2 gene amplification by fluorescence in situ hybridization and its diagnostic value in low-grade osteosarcoma].

Objective: To investigate the diagnostic value of detecting MDM2 gene amplification by fluorescence in situ hybridization (FISH) in low-grade osteosarcoma (LGOS). Methods: Thirty cases of parosteal osteosarcoma (POS) and 14 cases of low-grade central osteosarcoma (LGCOS) from April 2009 to August 2022 at Beijing Jishuitan Hospital, Capital Medical University were analyzed for the presence of MDM2 gene amplification by FISH. Fifty-eight additional cases were used as negative controls (including 28 cases of fibrous dysplasia, 5 cases of giant cell tumor, 4 cases of conventional osteosarcoma, 2 cases each of periosteal osteosarcoma, reparative changes after fracture, pleomorphic undifferentiated sarcoma, low grade myofibroblastic sarcoma, fibrous dysplasia with malignant transformation, one case each of leiomyosarcoma, sclerosing epithelioid fibrosarcoma, malignant peripheral nerve sheath tumor, desmoplastic fibroma of bone, solitary fibrous tumor, aneurysmal bone cyst, clear cell chondrosarcoma, osteofibrous dysplasia, and 3 cases of unclassified spindle cell tumor). Results: Among the 30 patients with POS, 15 were male and 15 were female, ranging in age from 10 to 59 years (mean 35 years, median 30.5 years). Among the 14 patients with LGCOS, four were male and 10 were female, ranging in age from 15 to 56 years (mean 37 years, median 36 years). All except one case were successfully detected by FISH. MDM2 gene amplification was detected in 27 cases of POS (27/29,91.3%) and 8 cases of LGCOS (8/14). All the negative controls were negative for MDM2 gene amplification. The positive rate of MDM2 gene amplification was significantly different between the case group and the control group (P<0.05). The sensitivity and specificity of MDM2 gene amplification in diagnosing POS and LGCOS were 91.3% and 100.0%; and 57.1% and 100.0%, respectively. The sensitivity and specificity of MDM2 gene amplification in diagnosing LGOS (including POS and LGCOS) were 81.3% and 100.0%, respectively. In cases where MDM2 gene was amplified, the MDM2 amplified signal was clustered. Nine cases showed increased CEP12 signal different from polyploidy which was displayed as small and weak signal points or cloud flocculent and cluster signals. Conclusions: Detection of MDM2 gene amplification by FISH is a highly sensitive and specific marker for LGOS. The interpretation criteria for FISH detection of MDM2 amplification are currently not unified. The signal characteristics need more attention when interpreting.

[Pediatric myofibroma/myofibromatosis of the soft tissue and bone: a clinicopathological analysis of 28 cases].

Objective: To investigate the clinicopathological features, diagnosis and differential diagnosis of pediatric myofibroma/myofibromatosis of the soft tissue and bone. Methods: All cases of pediatric myofibroma/myofibromatosis of the soft tissue and bone diagnosed between January 2011 and December 2018 were retrieved from the surgical pathology records in the Department of Pathology, Beijing Jishuitan Hospital, Beijing, China. Clinical and radiological data were collected. H&E and immunohistochemistry were used to examine histological and immunophenotypic features and to make the diagnosis and differential diagnosis. The relevant literature was also reviewed. Results: Twenty-eight cases of pediatric myofibroma/myofibromatosis of the soft tissue and bone were respectively collected. The patients' ages ranged from 2 months to 14 years, with a mean age of 7 years. There were 7 females and 21 males. There were 12 cases located in soft tissue, including the finger (n=9), upper arm (n=1) and foot (n=2). There were 14 cases located in the bone of limb, including the femur (n=8), tibia (n=4), clavicle (n=2), fibula (n=2) and radius (n=1). There were 2 cases of myofibromatosis involving multiple bones. Radiology showed lytic lesions in the bone. The proliferation of spindle-shaped myofibroblasts arranged in fascicles with indistinct eosinophilic cytoplasm and bland nuclei, with no pleomorphism and cytological atypia. The characteristic histologic structure was the biphasic nodular growth pattern with cellular and paucicellular regions. The tumors might arrange in a hemangiopericytoma-like pattern. The stroma varied between dense fibrosis and myxoid changes. The reactive new bone formation and inflammatory cell infiltration also existed. Immunohistochemical study showed that the SMA was positive. The surgical resections were performed. One of the patients had tumor recurrence as a result of 11-month follow-up. Conclusions: The pediatric myofibroma/myofibromatosis of the soft tissue and bone is a very rare benign tumor and has a good prognosis. It has a characteristic morphology and its differential diagnosis from other spindle cell tumors could be made with the immunohistochemical analysis.

Determination of the energy value of corn distillers dried grains with solubles containing different oil levels when fed to growing pigs.

This experiment used indirect calorimetry to determine the net energy (NE) content of five corn distillers dried grains with solubles (corn DDGS) containing different oil levels and to compare the NE obtained using indirect calorimetry with that calculated using previously published prediction equations. There were two samples of high-oil DDGS, one sample of medium-oil DDGS and two samples of low-oil DDGS. Twelve barrows (initial BW of 32.8 ± 2.0 kg) were used in a repeated 3 × 6 Youden square design with three periods and six diets. The diets were comprised of a corn-soybean meal basal diet and five diets containing 29.25% of one of the corn DDGS added at the expense of corn and soybean meal. During each period, the pigs were individually housed in metabolism crates for 16 days which included 7 days for adaption to feed and environmental conditions. On day 8, the pigs were transferred to respiration chambers and fed one of the six diets at 2300 kJ ME/kg BW0.6 /day. Faeces and urine were collected from day 9 to 13 and heat production (HP) was also measured. From day 14 to 15, the pigs were fed 893 kJ ME/kg BW0.6 /day to allow them to adapt from the fed to the fasted state. On the last day of each period (day 16), the pigs were fasted and fasting HP was measured. The digestible energy value was 16.0, 17.1 and 15.3 MJ/kg DM, the metabolizable energy value was 14.6, 15.5 and 13.7 MJ/kg DM and the NE value was 10.7, 11.0 and 9.4 MJ/kg DM, for the high-oil, medium-oil and low-oil corn DDGS, respectively. The NE obtained with indirect calorimetry in the present study did not differ from values calculated using previously published prediction equations.

Coimmunization of Agaricus blazei Murill extract with hepatitis B virus core protein through DNA vaccine enhances cellular and humoral immune responses.

DNA vaccines induce protective humoral and cell-mediated immune responses in several animal models. Agaricus blazei Murill (ABM) is particularly rich in polysaccharides, and has shown particularly strong results in treating and preventing cancers. The goal of this study was to investigate whether co-immunization of the fungus ABM with hepatitis B virus (HBV) core DNA vaccine could increase the immune responses. Compared with the control mice which received hepatitis B virus core antigen (HBcAg) alone, significant increase in not only the HBcAg-specific antibody response but also T cell proliferation was observed in mice which received HBcAg DNA vaccine plus ABM extract. These results suggest that ABM extract might represent an adjuvant to improve the efficacy of DNA vaccines in vivo.

Selective CD4+ lymphopenia in melanoma patients treated with temozolomide: a toxicity with therapeutic implications.

PURPOSE: Standard schedule temozolomide (TMZ; daily for 5 days every 4 weeks) is often used in melanoma patients, but phase III data show that it is no more effective than standard dacarbazine. Extended TMZ dosing regimens may be superior by delivering the drug continuously at a higher dose over time. Using an extended dosing schedule, we noted a high incidence of lymphopenia and occasional opportunistic infections (OIs). Here we report our retrospective experience in the first 97 patients. MATERIALS AND METHODS: TMZ was administered at 75 mg/m(2)/d orally for 6 weeks every 8 weeks, although nine patients were treated continuously without a break. Seventeen patients were treated with TMZ alone; 73 patients received TMZ with thalidomide; seven patients received TMZ with low-dose interferon alfa. RESULTS: Median duration of TMZ treatment was 113 days; 29% received > or = 24 weeks of therapy. Lymphopenia was seen in 60% of patients (absolute lymphocyte count < 800/microL) with a median of 101 days to lymphopenia. TMZ did not cause significant neutropenia or thrombocytopenia. Lymphopenia was not more common in patients treated concomitantly with thalidomide. In all patients analyzed for lymphocyte subsets, lymphopenia induced by TMZ affected the CD4(+) compartment preferentially. There were two documented OIs (Pneumocystis and Aspergillus pneumonia) as well as other infections indicative of T-cell dysfunction in another 21 patients. CONCLUSION: TMZ at this dose and schedule results in CD4(+) lymphopenia in a majority of patients that can result in OIs. Pneumocystis pneumonia prophylaxis should be considered for patients who develop sustained lymphopenia on TMZ.

Effects of vitamin E on the blastogenic response of splenocytes and lipofuscin contents in the hearts and brains of aged mice.

Semipurified diets containing 30 or 500 ppm of dl-alpha-tocopherol (VE) were fed for 12 weeks to young (3-month-old) and old (20-month-old) Swiss mice. We measured the blastogenic response of splenocytes, the serum VE, and the lipofuscin levels in brains and hearts. We found that old mice fed with 500 ppm VE diet had a significantly higher serum VE level and blastogenic response of splenocytes to concanavalin-A (ConA) and lipopolysaccharide (LPS) than those fed with 30 ppm VE diet (p < 0.01). However, the lipofuscin level in the brains and hearts of aged mice declined substantially with the VE supplementation (heart: p < 0.001, brain: p < 0.05). Furthermore, the effects of dietary VE on the serum VE and tissue lipofuscin content in aged mice were much more obvious than in the young animals.

Effects of vitamins and methionine on blastogenic response of splenocytes and lipid peroxidation in lung and liver of mice exposed to SiO2 dust.

We studied the effects of nutrients such as vitamin C, E, B6, niacin, and methionine on the blastogenic response of splenocytes and lipid peroxidation (LPO) in lung and liver of mice exposed to SiO2 dust. The results showed that after treatment with SiO2, the blastogenic response of splenocytes decreased significantly (p < 0.001), whereas the LPO level in lung and liver increased remarkably (lung p < 0.05; liver p < 0.001). Vitamin E, C, and niacin strongly enhanced the blastogenic response of T and B splenic lymphocytes to concanavalin-A and to lipopolysaccharide (p < 0.001). The effects of methionine and B6 were much less intense. Other supplied nutrients have been shown to inhibit LPO in lung and liver of mice exposed to SiO2 (p < 0.001).

Effects of dietary supplementation on the function of alveolar macrophages of silicosis rats, on the blastogenic response of lymphocytes and on the peroxidase activity in blood of silicosis patients.

We studied the effects of a nutrient supplemented diet on the function of alveolar macrophages of silicosis rats and on the blastogenic response of lymphocytes, glutathione peroxidase (GSH-PX) activity, and lipid peroxidase (LPO) activity in the blood of silicosis patients. The results showed that a nutrient supplemented diet increased the phagocytosis rate (p < 0.01) and index (p < 0.05) of alveolar macrophages of silicosis rats. A nutrient supplemented diet also enhanced significantly the GSH-PX activity (p < 0.001) and the blastogenic response of lymphocytes (p < 0.01), and decreased substantially the LPO content (p < 0.05) in the blood of silicosis patients. We conclude that a nutrient supplemented diet may play an important role in antilipid peroxidation, decreased free radical reaction, stabilizing cell membrane, delaying lung fibrosis, and enhancing immune functions of the body.