Adolescent ; Child ; Child, Preschool ; Female ; Homeodomain Proteins ; genetics ; Humans ; Infant ; Male ; Oncogene Proteins, Fusion ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

Objective To detection the methylation of p16INK4A in primary hepatocellular carcinoma, a nested bisulfite sequencing and methylation-specific polymerase chain reaction (BS-MSP) protocol was designed and used.Methods Bisulfite-modified DNA were amplified to evaluate the quality of templates with a pair of bisulfite sequencing primers in the first round of PCR, then subjected to methylation assay with corresponding methylation or unmethylation specific PCR primers.Representative PCR products were sequenced to confirm its correctness.Results 3 of 40 cases (7.5%) were failed to assay due to poor quality of templates, and 29 of 37 cases (78%) were detected p16INK4A methylation.Sequencing results confirmed that templates were correctly amplified.Conclusion BS-MSP technique might be valuable for methylation study on carcinogenesis and clinical assay.

Objective To observe the clinical effect of Blade needle closed release and massage for the patients with cervical heart syndrome. Methods A total of 60 patientswith cervical-heart syndrome were randomly divided into the control group and the treatment groups, 30 patients in each group. Both groups were treated with rehabilitation exercises, and the treatment group were added with Blade needle closed release and massage, while the control group were added with traction. The treatments were operated once to 3 times based on the patients' condition with an interval of seven days. The symptoms, signs and effective rates were observed and compared. Results After treatment, the neck and shoulder pain (0.7 ± 0.6 vs. 0.9 ± 0.7, t=2.904), precordialgia (0.5 ± 0.5 vs. 1.0 ± 0.7, t=4.210), chest tightness (0.5 ± 0.6 vs. 0.8 ± 0.7, t=2.093), cardiopalmus (0.7 ± 0.5 vs. 1.0 ± 0.8, t=3.915), spontaneous (0.7 ± 0.5 vs. 0.8 ± 0.9, t=2.837), vertigo (1.0 ± 0.6 vs. 1.6 ± 0.8, t=2.760) scores in the treatment group were significantly lower than those in the control group (P<0.05 or P<0.01). The total effective rate of the treatment group was 93.89% (28/30), and the Control group was 66.67%(20/30). There are significantly different(χ2=4.547, P<0.05). Conclusions The method of Blade needle closed release and traction could improve the symptom of the shoulder neck and heart area of the cervical-heart syndrome.

Objective To compare the effect of proximal femoral intramedullary nail fixation and femoral retrograde intramedullary nail combined with hollow lag screw fixation in the treatment of ipsilateral femoral neck and shaft fractures.Methods Data of 42 patients with ipsilateral femoral neck and shaft fractures who were admitted in our hospital from January 2007 to January 2016 were retrospectively analyzed.According to different fixation methods,the patients were divided into two groups:single intramedullary nail group (SIN group) (20 cases,treated with single proximal femoral intramedullary nail,15 males and 5 females with an average age of 39.4);combined internal fixation group (CIF group) (22 cases,treated with femoral distal intramedullary nail combined with femoral neck hollow lag screw,17 males and 5 females with an average age of 42.2 years).In the SIN group,4 cases were type A,9 cases type B,7 cases type C according to AO classification.And 14 cases belonged to Ⅰ or Ⅱ (stable fracture),6 cases of type Ⅲ or Ⅳ (unstable fracture) according to Garden classification.In the CIF group,3 cases were type A,10 cases type B,9 cases type C according to AO classification.And 16 cases belonged to type Ⅰ or type Ⅱ,6 cases type Ⅲ or Ⅳ according to Garden classification.The incision length,operation time,intraoperative blood loss,fracture healing time,postoperative weight time,stability of internal fixation,hip function score,pain in the knee and postoperative complication rate were compared between the two groups.Results There was no significant difference in gender,age and fracture classification between the two groups.The operation time (75.0±10.2 min),intraoperative blood loss (150.6±80.4 ml),hip Harris score excellent rate (80％) of incidence of knee pain in postoperative 3 months (10.0％) of the SIN group were significantly lower than those of the CIF group (105.2±18.4 min,180.0±56.8 ml,86.4％ and 31.8％).The length of incision (8.3±1.4 cm) in the SIN group was significantly longer than that in the CIF group (4.0±0.6 cm).There was no significant difference in fracture healing time (SIN group 20.O±4.0 weeks,CIF group 19.6±4.2 weeks) and postoperative weight time (SIN group 8.2±4.0 weeks,CIF group 8.0±4.2 weeks) between the two groups.The main complications of the two groups were delayed union (3 cases of SIN group and 1 case of CIF group),nonunion of fracture (1 case of SIN group) and,coxa vara (1 case of SIN group).The complication of SIN group (30％) was significantly higher than that of CIF group (4.5％),which occurred mainly in unstable femoral neck fracture cases.Conclusion Both proximal femoral intramedullary nail fixation and femoral retrograde intramedullary nail combined with hollow lag screw can effectively treat femoral ipsilateral femoral neck and shaft fracture.Femoral retrograde intramedullary nail combined with hollow lga screw has more therapeutic advantages for unstable femoral neck fractures (Garden Ⅲ and Ⅳ).

Carcinoma, Medullary ; complications ; pathology ; Humans ; Male ; Middle Aged ; Thyroid Neoplasms ; complications ; pathology

Objective To investigate the relationship between raf kinase inhibitor protein (SKIP), a novel metastasis suppressor gene, and metastasis of ovarian carcinoma. Methods Immunohistochemistry, RT-PCR, and western blot analysis were performed to examine the expression of SKIP in clinical samples of ovarian tumors and five human ovarian carcinoma cell lines. Stable cell lines over-expressed or deleted of SKIP were cloned to investigate the function of SKIP in ovarian cancer cells. The recombinant plasmids expressing sense (ss) or antisense (as) SKIP cDNA or empty vector was transfected into ovarian cancer cell line SKOV3 by lipofectamine. The expression level of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK) and extracellular signal-regulated kinase (ERK) in ovarian cancer cells were detected by western blot analysis. Assays of cell proliferation, soft-agar colony formation, cell adhesion, and cell invasion in vitro were used to examine the malignant phenotypes of the transfected cells. Flow cytometric analysis was performed to observe the effect of SKIP on cell cycle distribution before and after transfection. Results (1 ) The expression levels of SKIP protein in ovarian carcinoma tissues from patients were found to be reduced than those in ovarian benign tumor and borderline tumor. SKOV3 clones stably expressing full-length recombinant ssRKIP, asRKIP, and their respective empty vector were obtained. (2)RKIP was able to block basal levels of MEK and ERK in ovarian cancer cells. The expression level of phosphorylation MEK in ssRKIP#1 and ssRKIP#4 cells were 0. 35, 0. 34; while the expression level of phosphorylation ERK in ssRKIP#1 and ssRKIP#4 cells were 0.48 and 0.46. (3) Abilities of cell proliferation in the ssRKIP vector-transfected cells were decreased compared with that in the non-transfected cells (P ＜0. 01 ). (4)Anchorage-independent growth in the ssRKIP#1 and ssRKIP#4 cells (83.7 ± 5.7, 106. 0±9. 2) were decreased compared with that in the empty vector-transfected cells (158.3 ± 14. 6, P＜ 0. 01). (5)Cell adhesion in the ssRKIP#1 and ssRKIP#4 cells [(68.3±0. 8)%, (64. 1±0. 9)%] were decreased compared with that in the non-transfected cells [(100. 0 ± 1.1 )%, P ＜ 0. 01]. (6) Cell invasion in the ssRKIP#1 and ssRKIP#4 cells (24 ± 5, 25±4) were decreased compared with that in the non-transfected cells (68 ± 5, P ＜ 0. 01 ). (7) ssRKIP cells had a significant increase in the G1 phase and decrease in the G2 + S phase. Conclusion RKIP could inhibits the metastasis, but also the growth of ovarian cancer cells.

Objective To study the expression of macrophage inflammatory protein 2(MIP-2) and the interfering effects of naloxone in the brain edema caused by lioposacchride (LPS)in rats.Methods Eithty-four SD rats were randomly divided into 3 groups:normal saline group(NS group,n=28) 0.2 mL normal saline was injected by carotid into each rat;LPS group(n=28) with 200 ?g LPS;naloxone interfering group(NAL group,n=28)1 mg/kg naloxone was intraperitoneally injected at 10 min,1,2,6,12 h and following LPS injected 2 h before decapitation.The content of MIP-2 and even blue(EB) in brain tissue were detected at different time point.The brain water content was measured by drying method.Results The content of water and EB in LPS group were significan higher than those in NS group(P

Object: To investigate the effects of Huaiqihuang Granule, a compound Chinese herbal medicine, on expressions of nephrin and podocin of slit diaphragm of glomerular podocytes in rats with adriamycin-induced nephrosis and to explore the mechanism in reducing the proteinuria. Methods: Twenty SD rats were randomly divided into five groups: control group, model group, glucocorticoid group, Huaiqihuang Granule group and Huaiqihuang Granule plus glucocorticoid group. The 24-hour urine was collected 7, 14, 21 and 28 days after adriamycin injection respectively to measure 24-hour urinary protein, and all rats were sacrificed after 28-day treatment. Pathological changes in renal tissues were observed under a light microscope and an electron microscope. Expressions of nephrin and podocin mRNAs in renal cortex were determined by real-time polymerase chain reaction, and protein levels of nephrin and podocin were detected by Western blotting. Results: (1) In the model group and the treatment groups, the level of urinary protein increased significantly from the 14th day. (2) Under the light microscope, inflammatory cells and slight fibroplasia were found in renal interstitium of the model group, but there were less inflammatory cells in renal interstitium in the intervention groups than in the model group. Under the electron microscope, 29 days after adriamycin injection, extensive fusion of foot processes was observed. (3) The expressions of nephrin and podocin were higher in treatment groups than in the model group. (4) Proteinuria level was negatively correlated with the expressions of nephrin mRNA and nephrin and podocin proteins. Conclusion: The above results indicate that Huaiqihuang Granule can maintain the integrity of the slid diaphragram in podocyte, alleviate the lesion of glomerular filtration membrane, and decrease the proteinuria by up-regulating the expressions of nephrin and podocin. Huaiqihuang Granule plus glucocorticoid maybe has better effects than glucocorticoid alone.

Objective To explore the prognostic value of B-type natriuretic peptide (BNP) for 28-day mortality of elderly patients with non-cardiac critical ill in emergency intensive care unit (EICU). Methods A total of 70 elderly non-cardiac critically ill patients (age≥60 years) in EICU were enrolled, and the blood samples were collected to detect BNP level after the patients' admission to EICU. After 28 days, the mortality was assessed. Results Twenty-two patients (31.4 %) died during 28 days observation, whose BNP levels were significantly higher than that of the survivors [ln BNP: (6.4 ± 1.2) ng/L vs. ( 5. 1 ± 1.5 ) ng/L, P＜ 0. 05] ; BNP level had an area under the receiver operating characteristic curve of 0. 759 (95% CI: 0. 636-0. 882, P＜0.05) for predicting mortality,and the optimal cut point of BNP was 342 ng/L (sensitivity 77.3%, specificity 68.7%).Conclusions BNP level could be a predictor for 28-days mortality for elderly non-cardiac critically ill patients.

Adult ; Embolism, Amniotic Fluid ; etiology ; therapy ; Female ; Humans ; Pregnancy