AIM: To observe the expression of HIF-1? mRNA, HIF-1?, VEGF and iNOS proteins and to investigate their relationship in h ypoxia-treated SW480 cells. METHODS: HIF-1?, VEGF and iNOS proteins were measured by immuno cytochemistry. Western-blot was used to detect HIF-1? protein. HIF-1? mRNA wa s measured by in situ hybridization. RESULTS: Under hypoxic condition, SW480 cells expressed proteins of HIF-1?, VEGF and iNOS more strongly than that under normoxia condition. How e ver, under hypoxia condition, these three proteins expressed weakly or negativel y when the cells treated with genistein, the inhibitor of HIF-1?. Expressions o f HIF-1? and VEGF proteins in cultured SW480 cells under hypoxic condition were completely or partially inhibited by the addition of SNP but the expression of iNOS was unaffected. Another NO donor NOC5, however, induced the expression of t hese three proteins. L-NAME, a non-specific inhibitor of NOS, inhibited the expr ession of HIF-1?, VEGF and iNOS. The levels of HIF-1? mRNA changed slightly i n different oxygen condition or addition of genistein, NO donor or iNOS inhibitor . CONCLUSIONS: Hypoxia induces the expression of HIF-1?, therefor e upregulates the production of VEGF and iNOS. During hypoxia, SNP inhibits but N OC5 promotes HIF-1? expression, indicating that different NO donor acts on the cells through different mechanisms.

Objective To evaluate of impact of thalidomide on CD4 +CD25 +T lymphocytes in patients with acute myeloid leukemia and its clinical curative effect.Methods 71 cases of patients with AML were randomly divided into the control group and the experiment group, and 31 healthy people were selected to be the normal group.The experiment group and the control group patients were treated with the same chemotherapy, the experiment group was treated with thalidomide.The levels of CD4 +CD25 +T lymphocyte, CD3 +T lymphocyte, CD4 +T lymphocyte, CD8 +T lymphocyte, ratio of CD4 +/CD8 +and NK cells were detected at before treatment, 10 to 14d after treatment, complete remission 6 months follow-up.In normal group, the same index was detected before and after chemotherapy, and 10-14 d.The clinical curative effect of the experiment group and the control group were observed.Results The effective rate of the experiment group was higher than of the control group(P<0.05); The levels of CD4 +CD25 +T lymphocyte in the experiment group and control group were significantly higher than in the normal group(P<0.05), the two groups decreased compared with the control group, the level of CD4 +CD25 +T lymphocyte in the experiment group was significantly decreased(P<0.05).And with thalidomide treatment, the experiment group in the CD3 +T lymphocytes, CD4 +T lymphocytes and ratio of CD4 +/CD8 +, NK cells were significantly higher than the control group ( P<0.05 ) .Conclusion Thalidomide can improve both the immunity cell function and the clinical efficiency in patients with AML.The mechanism is related to reduce the level of CD4 +CD25 +T lymphocyte.

ObjectiveTo investigate the changes of serum concentration of parathyroid hormone (PTH) and calcium after thyroid surgery and compare the changes among different modes of operation. MethodsFrom Aug. 2006 to Dec. 2009, 470 patients accepted thyroid surgery. The serum concentration of PTH and calcium in different groups was measured and compared before and 1 day after surgery. According to the extent and similarity of the surgery, patients were classified into 7 groups and they were compared in terms of postoperative changes of PTH and serum calcium. Statistical analysis was performed. ResultsThe serum concentration of PTH and calciurn decreased significantly after surgery in all patients except for those receiving unilateral and bilateral partial thyroidectomy. Compared with unilateral lobectomy, surgeries such as bilateral subtotal thyroidectomy, unilateral thyroidectomy with contralateral subtotal thyroidectomy, bilateral near-total thyroidectomy and total thyroidectomy resulted in more dramatic decreases of serum concentration of PTH and calcium and higher incidence of hypocalcemia ( P ＜ 0.05 ). The comparison between patients receiving CLND or not had the same result. Conclusions Almost all kinds of thyroid surgery affect the parathyroid function. The wider the surgery, the higher the possibility of postoperative hypoparathyroidism. The indications and criteria of different types of thyroid surgery are essential for hypoparathyroidism prevention. In some cases, vitamin D and calcium are recommended for preventive purpose.

OBJECTIVETo explore the genetic causes for a child with multiple congenital malformations and epilepsy through analysis of copy number variations, and to correlate the genotype with the phenotype.

METHODSG-banding karyotyping was performed on the child and her parents. Single nucleotide polymorphisms array (SNP-array) was used to map the exact chromosomal breakpoints in the proband. The result was validated with fluorescence in situ hybridization (FISH).

RESULTSG banding analysis suggested that the proband had a karyotype of 46,XX,del(4)(p15), while both of his parents had a normal karyotype. SNP-array has identified a hemizygous deletion of 13.3 Mb on chromosome 4p16.3p15.33, which has been implicated in Wolf-Hirschhorn syndrome. FISH assay has confirmed the de novo origin of the deletion, with the karyotype and clinical phenotype of both parents taken into consideration.

CONCLUSIONA case of Wolf-Hirschhorn syndrome has been diagnosed by clinical manifestation and karyotyping analysis. Compared with conventional karyotyping analysis, SNP-array has greater resolution and accuracy, and can provide useful information for genetic counseling.

Chromosome Banding ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Infant, Newborn ; Karyotyping ; Oligonucleotide Array Sequence Analysis ; Polymorphism, Single Nucleotide ; Wolf-Hirschhorn Syndrome ; genetics

Objective: To investigate the effect of 2, 2', 4, 4'-tetrabromodiphenyl ether (PBDE-47) on the mitochondrial mass in rat adrenal pheochromocytoma (PC12) cells and the potential mechanisms. Methods: Highly differentiated PC12 cells were divided into control, 1, 10 or 20 μmol/L PBDE-47-treated groups and cultured for 24 h. Transmission electron microscopy was employed to observe the changes in mitochondrial morphology and quantity in PC12 cells. Flow cytometry was used to measure the fluorescence intensity of Nonyl Acridine Orange (NAO) , a fluorescent indicator of mitochondrial membrane cardiolipin, to reflect mitochondria mass. Western blotting was used to determine the expression levels of Mitofusion 1 (Mfn1) and Fission 1 (Fis1) proteins. To further explore the role of abnormal mitochondrial fusion and fission in PBDE-47-induced mitochondrial mass changes, PC12 cells were divided into control group, 5 μmol/L M1 treatment group, 20 μmol/L PBDE-47 treatment group and 5 μmol/L M1+20 μmol/L PBDE-47 combined treatment group and cultured for 24 h, then the fluorescence intensity of NAO and expression levels of Mfn1 and Fis1 proteins were detected. Results: The control group showed numerous mitochondria with normal morphology, while the number of mitochondria decreased after PBDE-47 treatment. Especially, the disappeared cristae, swelling and vacuoles of mitochondria and decreased fluorescence intensity of NAO (P<0.05) were observed in 10 and 20 μmol/L PBDE-47-treated groups. Meanwhile, the expression levels of Mfn1 and Fis1 proteins in the 10 and 20 μmol/L PBDE-47-treated groups were significantly decreased compared with control group (P<0.05) . However, 5 μmol/L M1 co-treatment with 20 μmol/L PBDE-47 significantly increased the levels of Mfn1 and Fis1 proteins and fluorescence intensity of NAO compared with the 20 μmol/L PBDE-47 group (P<0.05) . Conclusion PBDE-47 can inhibit the mitochondrial fusion and fission process, thus leading to damage of mitochondria mass in PC12 cells.

Objective To investigate the shedding of CAV2-ΔE3-CGS after immunization and the background of canine adenovirus (CAV) infection, and to establish a dual fluorescent quantitative PCR detection method for rabies virus (RV) and canine adenovirus type 2 (CAV2). Methods A dual fluorescent quantitative PCR detection method was established by designing specific primers and probes for E1 gene of CAV and G gene of RV for the detection of CAV2-ΔE3-CGS. Oral swabs, anal swabs and environmental samples of stray dogs from experimental animal farm and dog detention center were tested. Results The standard curves generated by this method were Y=-3.351 × logX + 44.895, R² = .999 and Y=-3.413 × logX + 45.192, R²=0.996, respectively. The linear relationships were good, and the minimum detection limits were both 102 copies/μL. CAV2-ΔE3-CGS was not detected in experimental animal farm. CAV was detected in dog detention center, and the positive rates were 5.88% (5/85) in oral swabs, 8.24% (7/85) in anal swabs, and 4% (1/25) in environmental samples. Conclusion The dual fluorescent quantitative PCR method can be used for the detection of CAV2-ΔE3-CGS after immunization and the investigation of CAV infection. The present study has shown that no CAV2-ΔE3-CGS has been detected after immunization and CAV infection rate of stay dogs is low in Shanghai. CAV2-ΔE3-CGS oral immunization meets requirement and is applicable.