Objective To explore the relationship between the stress during pregnancy and the NMDA-induced spasm. MethodsThe pregnant rats were divided into two groups: pregnant control group(PC group)and pregnatal stress group(PS group). The rats in PS group were forced to swim in cold water of 4℃ for five minutes. Pups of the two groups were injected NMDA 7mg/kg intraperitoneally on postnatal day 13 to induced infantile spasm-like attacks. Then the clinical behavior of pups of the two groups were observed and recorded. The ultrastructure of mitochondria was detected with scanning electron and appearance of neurons was observed with HE staining. NMDA receptor and its subuniforms (NR2A, NR2B) in hippocampus, cerebral cortex, hypothalamus and raphe nucleus of brainstem were estimated by immunohistochemistry. Results The average score of pups in PS group is higher than in PC group(7.84 ±1.01 vs 5.90 ± 1.12). There were swelling neurons and no cytonecrosis in hippocampuss of both two groups. The uhrastructure of mitochondria in hippocampuss and raphe nucleus of brainstem showed great changes in pups of PS group as membrane in mitochondria of pups in PS group. NR1, NR2A and NR2B distributed mainly on the membrane and also in cytoplasm and cellular nucleus. The NR1 expressions in hippocampus, cerebral cortex, hypothalamus and raphe nucleus in pups of PS group were elevated than those of PC group and the P value is 0.001, 0.009, 0.015, 0.001 respectively. The NR2B expressions in hippocampus, cerebral cortex hypothalamus and raphe nucleus in pups of PS group were elevated than those of PC group and the P value is 0.004, 0.002, 0.003, 0.002 separately. NR2A expressed more in hippocampus in pups of PS group than that in PC group,but there was no difference in cerebral cortex, hypothalamus and raphe nucleus between two groups and the P value is separately 0.006,0.067, 0.7, 0.5. Conclusions The mitochondria of neurons in rat pup were hurt by maternal severe stress daring pregnancy, which made the energy metabolism of neurons changed. Maternal stress can also elevate the expression of NR1, NR2A and NR2B in cerebral cortex, brainstem and hippocampus and increase the morbidity of infantile spasm-like attacks.

Objective To study the clinicopathological and immunohistochemical features , histogenesis and prognosis of pleuropulmonary blastoma ( PPB) in children.Methods PPB specimens from 16 pediatric cases with an age ranging from 1 year and 7 months to 5 years and 3 months ( mean age of 3 years) were retrieved and analyzed by routine histological , immunohistochemical and electron methods.Results Among 16 patients, there were 2 type Ⅰ, 7 type Ⅱ and 7 type Ⅲ PPB cases.Type Ⅰ PPB as multilocular cystic structure , consisted of thin fibrous wall lining the respiratory epithelium , subepithelial primitive blastema or immature mesenchymal cells , with or without rhabdomyoblastic differentiation or cartilage;Type Ⅱ PPB as cystic-solid tumor,comparing with type Ⅰ, consisted of intracystic components with appearance of anaplastic tumor cells.TypeⅢPPB consisted of completely solid mass , the same as the solid region of type Ⅱ, had mixed pattern including blastema ,undifferentiated spindle-cell proliferations and sarcomas.In addition, anaplastic tumor cells and intra-and extra-cytoplasmic eosinophilic globules were also commonly present.Epithelial components in PPB were benign.Immunohistochemical study showed primitive mesenchymal differentiation of tumors.All cases were positive for vimentin , desmin, myogenin and SMA in tumors with skeletal muscle differentiation , S-100 was positive in tumors with cartilage differentiation.All tumors were negative for synaptophysin ,CD99,and CD117.Benign epithelial components were positive for AE1/AE3 and EMA.In 12 cases, electron microscopy revealed few organelles in the primitive mesenchymal cells and rich heterochromatin in mesenchymal cells , the latter also demonstrating cytoplasmic myofilament dysplasia.Nine cases had clinical follow-up ranging from 5 to 48 months, of which 4 patients died.Conclusions PPB is a rare lung neoplasm of children under the age of 6 years,with distinct pathological morphology .PPB may arise from lung or pleura mesenchymal cells and has a poor clinical outcome.

OBJECTIVETo study the clinicopathological and immunohistochemical features, histogenesis and prognosis of pleuropulmonary blastoma (PPB) in children.

METHODSPPB specimens from 16 pediatric cases with an age ranging from 1 year and 7 months to 5 years and 3 months (mean age of 3 years) were retrieved and analyzed by routine histological, immunohistochemical and electron methods.

RESULTSAmong 16 patients, there were 2 type I, 7 type II and 7 type III PPB cases. Type I PPB as multilocular cystic structure, consisted of thin fibrous wall lining the respiratory epithelium, subepithelial primitive blastema or immature mesenchymal cells, with or without rhabdomyoblastic differentiation or cartilage; Type II PPB as cystic-solid tumor, comparing with type I, consisted of intracystic components with appearance of anaplastic tumor cells. Type III PPB consisted of completely solid mass, the same as the solid region of type II, had mixed pattern including blastema, undifferentiated spindle-cell proliferations and sarcomas. In addition, anaplastic tumor cells and intra-and extra- cytoplasmic eosinophilic globules were also commonly present. Epithelial components in PPB were benign. Immunohistochemical study showed primitive mesenchymal differentiation of tumors. All cases were positive for vimentin, desmin, myogenin and SMA in tumors with skeletal muscle differentiation, S-100 was positive in tumors with cartilage differentiation. All tumors were negative for synaptophysin, CD99, and CD117. Benign epithelial components were positive for AE1/AE3 and EMA. In 12 cases, electron microscopy revealed few organelles in the primitive mesenchymal cells and rich heterochromatin in mesenchymal cells, the latter also demonstrating cytoplasmic myofilament dysplasia. Nine cases had clinical follow-up ranging from 5 to 48 months, of which 4 patients died.

CONCLUSIONSPPB is a rare lung neoplasm of children under the age of 6 years, with distinct pathological morphology. PPB may arise from lung or pleura mesenchymal cells and has a poor clinical outcome.

Child, Preschool ; Cysts ; pathology ; Desmin ; analysis ; Female ; Humans ; Infant ; Lung Neoplasms ; chemistry ; pathology ; Male ; Microscopy, Electron ; Myogenin ; analysis ; Prognosis ; Pulmonary Blastoma ; chemistry ; pathology ; Sarcoma ; pathology ; Vimentin ; analysis

Objective To construct programmed cell death protein-ligand 1(PD-LI)^hi tolerogenic dendritic cell (tol-DC) derived from bone marrow of DA rats and identify its immunological function. Methods DA rat bone marrow cells were extracted, combined with recombinant mouse granulocyte macrophage colony-stimulating factor and recombinant mouse interleukin (IL)-4, and cultured for 6 days in vitro to induce the differentiation of bone marrow cells into immature dendritic cells (imDC). Lipopolysaccharide was used to stimulate cell maturation and cultured for 2 days to collect mature dendritic cells (mDC). PD-L1 lentiviral vector virus stock solution or equivalent dose lentiviral stock solution was added, and PD-L1^hitol-DC and Lv-imDC were collected after culture for 2 days. The morphology of PD-L1^hitol-DC was observed by inverted phase contrast microscope and transmission electron microscope. Real-time fluorescence quantitative reverse transcription polymerase chain reaction, Western blotting and flow cytometry were used to detect the expression level of specific markers on cell surface. CD8⁺T cells derived from Lewis rat spleen were co-cultured with imDC, mDC, Lv-imDC and PD-L1^hitol-DC, respectively. The levels of inflammatory factors in the supernatant of each group were detected by enzyme-linked immunosorbent assay. The apoptosis of T cells and the differentiation of regulatory T cells (Treg) in each group were analyzed by flow cytometry. Results The morphology of PD-L1^hitol-DC modified by PD-L1 gene was consistent with tol-DC characteristics, and the expression levels of CD80, CD86 and major histocompatibility complex (MHC) on the surface were low. After mixed culture with CD8⁺ T cells, the levels of IL-10 and transforming growth factor (TGF) -β1 in the supernatant of PD-L1^hitol-DC group were higher, the levels of tumor necrosis factor (TNF) -α and IL-17A were lower, and the apoptosis of T cells and Treg differentiation were increased. Conclusions Overexpression of PD-L1 through lentiviral vectors may successfully induce the construction of bone-marrow derived PD-L1^hitol-DC in DA rats, promote the secretion of anti-inflammatory factors and T cell apoptosis, induce the differentiation of Treg, and inhibit the immune response of allogeneic CD8⁺T cells, which provides experimental basis for the next organ transplantation immune tolerance study.