Background and purpose:Multidrug resistance(MDR)is the main obstacle of chemotherapy in the treatment of cancer,and it has been reported that the muted p53 gene is related to MDR.In this study,we explored whether adenovirus mediated p53 gene(Ad-p53)could reverse MDR of human breast cancer and its impacts on the expressions of P-gp,TOPOⅡ and GST-?.Methods:In this study,adriamycin-resistant human breast carcinoma cells(MCF-7/Adr)and its parental cells(MCF-7)were used to determine the effect of Ad-p53.Cck-8 assay was adopted to evaluate the cytotoxicity of adriamycin.Western blot were performed to observe the expression of P-glycoprotein(P-gp),TopoismeraseⅡ(TOPOⅡ)and GST-?.Results:After transfection with 50 MOI Ad-p53,the 50% inhibitory concentration(IC50)of adriamycin to MCF-7/Adr cells was decreased from(4.54?0.91)?g/ml to(0.26?0.11)?g/ml,and the chemosensitivity increased 18.1 times(P

Objective To investigate the influence of difference ileocecal junction length on the efficacy of laparoscopic subtotal colectomy combined with antiperistaltic cecorectal anastomosis (LSCACRA) in treatment of slow transit constipation (STC). Methods Ninety- two STC patients undergoing LSCACRA were divided into 2 groups by random digits table method:10-15 cm group (10-15 cm ascending colon preserved above ileocecal junction, 46 cases) and 2-3 cm group (2-3 cm ascending colon preserved above ileocecal junction ,46 cases). All the patients were followed up for 24 months, and the Wexner incontinence score (WIS), ileocecal junction emptying time of barium enema, Wexner constipation score (WCS), abdominal pain intensity score (NRS), gastrointestinal quality of life index (GIQLI), abdominal pain frequency score and abdominal bloating frequency score were compared between 2 groups. Results All the patients successfully completed LSCACRA, with no conversion to open surgery and death occurred. There were no statistical differences in amount of bleeding, operative time, hospitalization time, exhaust time, incidences of postoperative intestinal obstruction and pulmonary infection, defecation time, WIS and abdominal bloating frequency score between 2 groups (P>0.05). The WCS, GIQLI, NRS and abdominal pain frequency scores 6, 12 and 24 months after operation in 2-3 cm group were significantly better than those in 10 - 15 cm group, WCS: (1.7 ± 1.3) scores vs. (4.2 ± 2.3) scores, (1.7 ± 1.1) scores vs. (4.1 ± 1.9) scores, (1.2 ± 0.5) scores vs. (3.9 ± 2.5) scores;GIQLI:(116.8 ± 6.2) scores vs. (98.5 ± 14.7) scores, (122.9 ± 7.5) scores vs. (104.7 ± 16.5) scores, (124.3 ± 5.9) scores vs. (108.3 ± 15.1) scores; NRS: (1.3 ± 0.5) scores vs. (2.5 ± 1.7) scores, (0.9 ± 0.3) scores vs. (2.3 ± 1.4) scores, (0.8 ± 0.3) scores vs. (2.2 ± 1.5) scores;abdominal pain frequency score:(0.9 ± 0.3) scores vs. (1.6 ± 1.2) scores, (0.7 ± 0.3) scores vs. (1.4 ± 1.1) scores, (0.7 ± 0.2) scores vs. (1.2 ± 1.0) scores, and there were statistical differences (P<0.05). The ileocecal junction emptying time of barium enema 24 months after operation in 2-3 cm group was significantly shorter than that in 10-15 cm group: (17.6 ± 8.4) h vs. (21.3 ± 10.8) h, and there was statistical difference (P<0.05). Conclusions STC patients with LSCACRA is safe and effective. The shorter length of ascending colon preserved above ileocecal junction can improve the efficacy of LSCACRA in the treatment of STC and the prognosis of patients. The 2-3 cm length of ascending colon preserved above the ileocecal junction should be recommended.

Pancreatic adenosquamous carcinoma is a rare exocrine pancreatic cancer,and surgical treatment is the only promising method in survival time extension and life quality enhancement.In July 2013,1 patient with pancreatic adenosquamous carcinoma and sarcomatoid change was admitted to the Shaoxing People's Hospital.A cauda pancreatis cystic nidus was detected by computed tomography 1 year ago,and no any other clinical symptoms were detected.The left upper quadrant of the patient was glutted and unwell,and the levels of CA19-9 and CA125 were 3 630 kU/L and 645 kU/L,respectively.The results of computed tomography and magnetic resonance imaging showed that a huge cystic and solid tumor was located between the cauda pancreatis,the stomach and the spleen;the tumor was mainly cystic and rooted in the tail of the pancreas;the spleen,stomach and surrounding tissues were infiltrated by the tumor.The results of postoperative pathological examination showed that the tumor was pancreatic adenocarcinoma with squamous epithelial differentiation and sarcomatoid change;the invasive squamous carcinoma dominated stomach wall,and the sarcomatoid change dominated spleen and lymph nodes with partial squamous epithelial differentiation and surrounding lymph nodes metastases.Although the surgical resection was complete and the postoperative thymosin injection and traditional Chinese medicine were implemented,tumor recurred at postoperative month 5,and the patient was dead due to cachexia and pernicious anemia at postoperative month 8.The prognosis of patients with pancreatic adenosquamous carcinoma is poor,early detection and surgery are key points for curative effect enhancement.

BACKGROUND:Both osteopontin and leptin are closely linked to bone metabolism, therefore, which may be related to the attack of femoral head necrosis. OBJECTIVE:To study the correlation of serum osteopontin and leptin in the development of femoral head necrosis. METHODS:Thirty-one patients with femoral head necrosis (case group, including 11 cases of ARCO II, 10 of ARCO III, 10 of ARCO IV) and 10 healthy adults (control group) were selected as the research objects. Enzyme-linked immunosorbent assay was used to determine the levels of osteopontin and leptin in serum folowed by statistical analysis. RESULTS AND CONCLUSION:In the case group, the serum levels of osteopontin and leptin were significantly higher than those in the control group (P < 0.05). But the serum levels of osteopontin and leptin showed no correlation between the two groups. These findings indicate that the serum levels of osteopontin and leptin are both elevated in patients with femoral head necrosis, which maybe play a role in the pathogenesis of femoral head necrosis, but there is no obvious correlation.

Objective To explore the effect of pathogenesis,diagnosis,therapy and prevention of portal vein thrombosis(PVT) after devascularization.Methods Data of 86 patients who underwent devascularization because of cirrhotic portal hypertension between January 2009 and December 2014 were retrospectively analyzed.Patients were divided into preventive anticoagulant treatment group (46 cases) and nonpreventive anticoagulant treatment group (40 cases) according to whether or not to receive prophylactic anticoagulant therapy.Compared the causes of PVT and clinical characters.Results The occurrence of PVT was 10.9％ (5/ 46) in preventive anticoagulant treatment group,PVT Ⅰ , Ⅱ, Ⅲ, Ⅳdegree were 2,3,0,0 cases,and that was 32.5％ (13/40) in nonpreventive anticoagulant treatment group, PVT Ⅰ , Ⅱ , Ⅲ, Ⅳ degree were 4,7,2,0 cases, the incidence rate of treatment group was lower than control group (x2 =9.735, P ＜ 0.05).There was no relationship in PVT with sex, age, and intraoperative ligation of splenic artery beforehand (P＞0.05).There were apparent correlation in PVT with centrifugal-flow of portal vein and peak value of platelet count(P＜0.05).All patients discharged uneventfully after anticoagulant treatment, including 2 cases of thrombolytic and 4 cases of blood supply recanalization, no case died from PVT.Conclusion The definite cause of PVT after devascularization is centrifugal-flow of portal vein and the peak value of platelet count (300-499) × 109/L is a pivotal risk factor.The key point of therapy is early diagnosis and early anticoagulant treatment.

Objective To investigate the effects of BMI-1 expression inhibition by RNA interference on the radiosensitivity of esophageal cancer TE-13 cells and its mechanism.Methods The siRNA based on the sequence of BMI-1 mRNA was synthesized to transfect cultured TE-13 cells as BMI-1 siRNA group,a negative one was synthesized to transfect cultured TE-13 cells as negative control group (NC group),and untransfected TE-13 cells were named as control group.The expression of the BMI-1 mRNA and protein in TE-13 cells was measured by quantitative real-time PCR and Western blot,respectively.The cell proliferation and the radiosensitivity of TE-13 cells were measured by MTS and colony-forming assay,respectively.Flow cytometry was used to analyze cell cycle and apoptosis.The expression of BCL-2 and BAX in TE-13 cells was measured by Western blot.Comparison between groups was made by analysis of variance.Results The BMI-1 siRNA group had significantly lower expression of BMI-1 mRNA and protein than the control group and the NC group (P=0.000,0.000).The proliferation of TE-13 cells in the BMI-1 siRNA group decreased significantly after irradiation (P=0.031).The colony-forming assay showed that the BMI-1 siRNA group had a significantly higher radiosensitivity than the control group and the NC group (P=0.000).After irradiation,the BMI-1 siRNA group had a significantly lower percentage of cells in G2/M phase than the control group and the NC group (P=0.000,0.000).The BMI-1 siRNA group had a significantly increased apoptosis rate (P=0.000,0.000),significantly reduced expression of BCL-2(P=0.000,0.000),and significantly increased expression of BAX after irradiation (P=0.000,0.000).Conclusions BMI-1 siRNA can inhibit the expression of BMI-1 gene in esophageal cancer TE-13 cells,eliminate the cell cycle arrest in G2/M phase,induce cell apoptosis after ionizing irradiation in vitro,and increase the radiosensitivity,which may be related to the regulation of the expression of BCL-2 and BAX.

BACKGROUND:There is evidence that internal fixation through an anterior or posterior approach for treatment of severe kyphotic deformity in spinal tuberculosis exhibits good curative effects. However, few prospective, long-term fol ow-up case control studies are reported. OBJECTIVE:To investigate the efficacy and safety of cobalt al oy pedicle screw implantation for treatment of severe kyphotic deformity in spinal tuberculosis. METHODS/DESIGN:This is a prospective, single-center, self-control, open-label trial, which wil be performed at the Affiliated Hospital of Hebei University, China. Eighty-four patients with severe kyphotic deformity in spinal tuberculosis wil be included according to the diagnosis criteria. Among 52 patients with tuberculosis of the thoracic spine, 28 wil undergo surgery through a posterior approach, and 24 through an anterior approach. According to the Frankel Grade classification, grade C, D and E spinal cord function wil be assessed in 8, 31 and 13 patients, respectively. Surgery through a posterior and anterior approach wil be respectively performed in half of 32 patients with tuberculosis of the lumbar spine. Grade C, D and E spinal cord function wil be assessed in 7, 14 and 11 patients, respectively. The primary outcome measure of this study wil be the Cobb angle at the thoracic spine segments before and 2 years after surgery, which wil be used to evaluate the angle of the spine curvature at the thoracic segments. The secondary outcome measures wil be X-ray scan or MRI findings before and 2 years after surgery, which wil be used to evaluate vertebral fusion after internal fixation;and Frankel Grade before and 2 years after surgery, which wil be used to evaluate recovery of spinal cord function after injury. Other outcome measures wil include multiple logistic regression analysis results of the factors that influence patient's curative effects and the incidence of adverse events 2 years after surgery. The trial protocol has been approved by the Ethics Committee, Affiliated Hospital of Hebei University, China and wil be performed in strict accordance with the Declaration of Helsinki, formulated by the World Medical Association. Signed informed consent regarding the trial protocol wil be obtained from each participant. DISCUSSION:This study is to validate that cobalt al oy pedicle screw implantation shows precise curative effects in the treatment of severe kyphotic deformity in spinal tuberculosis and to analyze through what approach, posterior or anterior, internal fixation wil be more beneficial to surgery performance. The outcomes of this study wil provide objective long-term fol ow-up evidence for internal fixation treatment of severe kyphotic deformity in spinal tuberculosis in the clinic.

Background and purpose:B cell-specific MLV integration site 1 (BMI-1) gene plays an important role in DNA damage after exposure to irradiation. The present study aimed to investigate the effect ofBMI-1 on radio-sensitivity of esophageal carcinoma cell after down-regulation of BMI-1 expression by silencing siRNA.Methods:Three pairs of siRNA based on the sequences of the BMI-1 mRNA were synthesized (siRNA1, siRNA2 and siRNA3) by compa-ny, and transfected into cultured TE13 cells as the BMI-1 siRNA groups, and a negative one was synthesized to be used as the negative control (NC) group. The untransfected group was named as the control group. BMI-1 mRNA and protein expression in esophageal cancer TE13 cells were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot in different groups. This study used flow cytometry assay to analyze cell cycle of transfected cells, and examined cellular growth and radiosensitivityin vitro by MTT and clone formation assay. mRNA and protein expression of p16 and CDK4 in esophageal cancer TE13 cells were detected by RT-PCR and Western blot.Results:The results of RT-PCR and Western blot showed that the expressions of BMI-1 at gene and protein levels were inhibited after silencing the BMI-1 gene. The mRNA and protein expression of BMI-1 in BMI-1 siRNA3 group were both significantly lower than that in BMI-1 siRNA1 and 2 groups. There was no significant difference in the cell proliferation among control, NC and BMI-1 siRNA3 groups. The values ofD0,Dq, and SF2 in BMI-1 siRNA3 group were 1.761, 2.122 and 0.6255, respectively, obvi-ously lower than those in control group (2.514, 2.694 and 0.8268) and those in NC group (2.506, 2.664 and 0.8231), while the value of N in BMI-1 siRNA3 group (3.336) was higher than that in control group (2.92) and that in NC group (2.895), which showed higher radiosensitivity in BMI-1 siRNA3 group. In addition, the cell cycle was arrested at G2/M phase after irradiation in control and NC groups. The percentage of G0/G1 phase in BMI-1 siRNA3 group was higher than that of control group and NC group, while the percentage of G2/M phase was lower than those in the latter. The up-regulation of p16 and down-regulation of CDK4 at gene and protein levels were detected after knockdown of BMI-1 expression by siRNA (P<0.01).Conclusion:siRNA could inhibitBMI-1 gene expression in esophageal cancer TE13 cells and enhance radiosensitivity, followed by eliminating the cell cycle arrest at G2/M stage after irradiationin vitro, which is related to the regulation of the protein expression ofp16 andCDK4.

Objective To develop a method of studying fiber projecting in the spinal cord duiring chicken embryo development.Methods At embryonic incubation 3 day (E3), pCAGGS-green fluorescent protein (GFP) plasmid was injected into the spinal cord using in vivo electroporation.Three days after transfection (E6), GFP-positive embryos were collected under a stereo fluorescence microscope .Subsequently , the spinal cord was separated from the embryos and cut from the roof plate as an open book .After fixed with 4%paraformaldehyde ( PFA) for one hour , the opened spinal cords were used for immunohistochemistry with N-cadherin antibody and with DAPI for nuclei .Finally, the nerve fiber projecting was photographed and analyzed under a fluorescence microscope . Results Based on the opened spinal cord and immunostaining in the cryosection , we observed that the nerve fibers projected across the midline of the floor plate and reached to the sulcus terminalis along the white matter of the contra side .The immunoreaction against N-cadherin indicated that overexpression of GFP has no significant effect on chicken embryonic development .Conclusion A new method to study fiber projecting in the developing chicken spinal cord is established successfully in this study .

A high-speed counter-current chromatography (HSCCC) method was successfully developed for the preparative separation and purification of deoxyschizandrin from Schisandrae Sphenantherae Fructus in one step. The purity of deoxyschizandrin was 98.5%, and the structure was identified by MS, UV and NMR. This method was simple, fast, convenient and appropriate to prepare pure compound as reference substances for related research on Schisandrae Sphenantherae Fructus.