Objective To investigate cell proliferation and invasiveness of cervical cancer HeLa229 cell after knockdown of NF-?B signaling pathway by P65 siRNA.Methods RNA interference was employed for specific inhibition of the expression of P65.HeLa229 cell was divided into transfected group and untransfected group.Cell viability was detected by MTT after the HeLa229 cells were transfected with or without P65 siRNA for 24,48,72 h.The sensitivity to 5-Fu of the HeLa229 cell,transfected with or without P65 siRNA,was evaluated also by MTT.Boyden chamber experiment in vitro was used to detect the invasion of HeLa229 cell.Results P65 siRNA inhibited the cell proliferation as compared with the untransfected cells.Proliferations of both cells transfected with and without P65 siRNA were inhibited in a concentration-dependent manner,while at the same concentration of 5-Fu the viability of transfected HeLa229 cells was significantly suppressed(P

To study the cartilage differentiation of mouse mesenchymal stem cells (MSCs) induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2 (0, 10, 20, 50 and 100 ng/mL). After 14 days of induction, morphology of cells was observed under phase-contrast microscope. Collagen II mRNA and protein were examined with RT-PCR, Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining. RT-PCR showed that CDMP-2 could promote expression of collagen II mRNA in an dose-dependant manner, especially at the concentration of 50 ng/mL and 100 ng/mL. Immunocytochemistry and Western blotting revealed a similar change. Alcian blue staining exhibited deposition of typical cartilage extracellular matrix. Our results suggest that mouse bone marrow mesenchymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the role of CDMP-2 in chondrogenesis.
Bone Marrow Cells/*cytology ; Bone Morphogenetic Proteins/*pharmacology ; Cell Differentiation/*drug effects ; Cells, Cultured ; Chondrocytes/*cytology ; Chondrogenesis/drug effects ; Chondrogenesis/physiology ; Mesenchymal Stem Cells/*cytology ; Recombinant Proteins/pharmacology

Objective Oral and Maxillofacial trauma Diagnosis and Treatment Expert system (OMDTES)was used in maxillofacial trauma PBL teaching to improve the quality of PBL teaching,and perfeet the assessment criteria, Methods OMDTES was applied in two procedures of maxillofacial injury PBL teaching activity:the preparation of lesson plan and assessment criteria.Then a questionnaire was designed to assess the effect of this new mode of PBL teaching.Results The new teaching model is welcomed by students and can improve students'interest in learning,the satisfaction of teaching and ameliorate method of assessment of PBL teaching.Conclusion Applying OMDTES in oral and maxinofacial injury PBL teaching has special advantages in raising the leavnevs'activity of learning,training their ability of chnical thinking and analysis.And it is worthy of further research and amelioration.

Objective To investigate the relationship between portal vein pressure and liver regeneration after 90% portal branch ligation in rats.Methods Forty-five male SD rats underwent 90% portal branch ligation (including 5 rats underwent sham operation),and then the changes of portal vein pressure and weight of unligated hepatic lobes were detected.The morphological changes of hepatocytes of the unligated hepatic lobes were observed under a light microscope.Proliferative cell nuclear antigen(PCNA)index was detected by immunohistochemistry,and hepatocyte apoptosis of the unligated hepatic lobes by TUNEL method.All data were analyzed by Pearson rank correlation analysis and t test.Results Thirty-eight out of 40 rats survived(95%).The ligated hepatic lobes diminished progressively,whereas the unligated hepatic lobes regenerated.Preoperative portal vein pressure was(9.1±1.8)cm H_2O(1 cm H_2O=0.098 kPa),and it was increased significantly shortly after the ligation and reached (15.8±2.7)cm H_2O 12 hours later(t=6.847,P＜0.05).The portal vein pressure decreased from(13.6±2.3)cm H_2O at day 1 to(9.3±2.0)cm H_2O at day 28.Preoperative positive PCNA index was 7%±3%,which was significantly lower than 14%±5%at postoperative 12 hours,21%±6%at day 3 and 26%±7%at day 5(t=9.129,P＜0.05),and it began to return to normal at day 5.Few apoptotic hepatoeytes were observed in preoperative liver tissue and unligated hepatic lobes.The expression of PCNA in unligated hepatic lobes and portal vein pressure had apositive correlation at postoperative day 1,3,5(r=0.913,0.896,0.908,P<0.05)and a negative correlation at postoperative day 14(r=-0.926,P<0.05).Conclusions The regeneration of hepatocytes in unligated hepatic lobes is activated after 90% portal branch ligation,and the regenerated liver compensates the weight loss of the ligated hepatic lobes.Liver is regenerated mainly by speeding hepatocyte proliferation rather than reducing hepatocyte apoptosis.Changes of portal vein pressure may play an important role in liver regeneration.

Objective To investigate the clinical value of emergent perspective stent implantation for leftside obstructive colonic cancer.Methods The clinical data of 26 patients with obstructive colonic cancer who received emergent perspective stent implantation at the Affiliated Xuzhou Hospital of Southeast University from October 2011 to February 2014 were retrospectively analyzed.A soft and a hard guidewire and a catheter were applied in the operation.The guidewires were put through the hole of the intestinal tumor,and then the metal stent and the conveyor were guided by the guidewires and were pulled through the hole of the intestinal tumor,finally the stent was released and the conveyor was adjusted to ensure that the stent was at the right position.Patients were followed-up through outpatient examination till 4 weeks after tumor resection.Results Of the 26 patients,9 were with rectal cancer,10 with sigmoid colonic cancer,6 with descending colonic cancer,1 with splenic flexure cancer.The median length of stenosis was 4.6 cm (range,2.0-8.0 cm).The surgery of the 26 patients was successful.The mean operation time was 35.2 minutes (range,15.2-72.0 minutes),and the mean time of stent implantation was 5.6 minutes (range,2.6-26.9 minutes).Patients had watery or loose stool for 4-8 times after stent implantation.Ten hours after the operation,all the patients were given liquid diet.The remission rate of clinical symptoms was 100.0％ (26/26).No colonic perforation was detected during the operation.Two patients were complicated with slight bleeding,and was alleviated by medication.Twenty-six patients received stage Ⅰ tumor resection procedure within 7-10 days after the symptoms of intestinal obstruction were remised.The success rate of surgery was 100.0％ (26/26).No infection and other drainage were detected after tumor resection through follow-up.Conclusion Emergent perspective stent implantation for left-side obstructive colonic cancer is safe and effective.

Background:CpG island methylator phenotype(CIMP)involving tumor suppressor gene( TSG)on short arm of chromosome 3(chromosome 3p)has been found in various types of cancers. However,its correlation with gastric cancer has not been clarified. Aims:To study the clinical significance of CIMP involving TSG on chromosome 3p in gastric cancer. Methods:Methylation specific PCR(MSP)was used to examine methylation profiles for hOGG1,VHL,RAR-B, hMLH1,SEMA3B,RASSF1A,BLU and FHIT harbored in chromosome 3p in 100 gastric cancer and paired paracancerous tissues. High CIMP( CIMP-H)was referred for those samples having four or more synchronously methylated genes. Relationship between CIMP-H and clinicopathological characteristics in gastric cancer was analyzed. Results:Positive methylation rates of VHL(P = 0. 030),hMLH1(P < 0. 001),SEMA3B(P = 0. 003),RASSF1A(P < 0. 001)and FHIT(P < 0. 001)were significantly higher in gastric cancer tissue than in paracancerous tissue. Incidence of CIMP-H rate in gastric cancer tissue was significantly higher than that in paracancerous tissue(44. 0% vs. 4. 0% ,P < 0. 001). CIMP-H was negatively correlated with degree of tumor differentiation(P = 0. 004),and positively correlated with lymph node metastasis(P = 0. 005),but not correlated with gender,age,tumor location,tumor size,depth of infiltration and TNM staging(P > 0. 05). Conclusions:CIMP on chromosome 3p may occur in early stage of oncogenesis of gastric cancer,and influencing tumor differentiation and lymph node metastasis.

Objective To investigate the expression of cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) in astrecytomas, as well as the correlation between them. Methods The expression of COX-2, EGFR and PCNA were respectively detected by immunohistochmical (S-P) method in 68 astrocytomas and 5 cases normal brain tissue. Proliferation index (PI) was calculated and the correlation of COX-2, EGFR and PI was analyzed. Results COX-2 and EGFR were negative expression in normal brain tissue. The positive expression rate of COX-2 and EGFR in high grade astrocytomas was significantly higher than that in low grade astrocytomas(73.53% vs 44. 18% ,67.65% vs 38.24%, P <0. 01 ), and the PI was significantly higher than that in low grade astrocytumas as well as normal brain tissue(46.11 ± 10. 68vs 23. 04±6. 25,4. 52±0. 95, P <0. 01 ). The PI in COX-2 positive group was higher than that in negative group( P <0. 01 ). The positive expression rate of COX-2 in the group with EGFR positive expression was higher than that in the negative group. Conclusions The expression of COX-2 and EGFR was related to pathological feature of astrocytomas. COX-2 may promote the proliferation of tumor cells. There was a static correlation between the expression of EGFR and COX-2 in astrocytomas. EGFR signal transduction probably modulated the expression of COX-2 in astrocytomas cells.

By preventing mesenchymal stem cells (MSCs) from adhering to precoated agarose to create a model of MSC suspension in vitro, we investigated anoikis in MSCs and the role of caspase-3 in the anoikis. The cultured MSCs were randomly divided into 3 groups: the anoikis group, caspase-3 inhibitor group and control group. Before experiment, we coated dishes with 1.5 % agarose; in the anoikis group, MSCs were put into the precoated dishes; and in the inhibitor group, caspase-3 inhibitor and MSCs were also put into the precoated dishes; but there were not intervention in the control group. MSCs were collected at 2 h, 6 h, 12 h and 24 h. The alteration of caspase-3 activity was evaluated by caspase-3 fluorometric assay and western blot analysis. The apoptosis rates were detected by flow cytometry. MSCs were round and suspended sufficiently in the anoikis and inhibitor groups. Caspase-3 fluorometric assay showed that there were significant differences in statistics between the anoikis group and the others (P<0.05). Western blot analysis discovered that caspase-3 expression in the anoikis group was more than that in the control and inhibitor groups (P<0.05). Flow cytometry showed that the apoptosis peak appeared in all the three groups, but it increased dramatically in the anoikis group. The apoptosis rates in the inhibitor and control groups were low and stable. And there were significant differences in statistics between the anoikis group and the others (P<0.05). MSCs will undergo anoikis in suspended condition if they are separated from the extracellular matrix. Caspase-3 inhibitors can suppress caspase-3 activity and reduce the apoptosis rate significantly. Caspase-3 plays a vital part in induced MSC anoikis in vitro. MSCs suspension culture system might be set up with argorose and caspase-3 inhibitor.

OBJECTIVE:To study the purification technology of 4 active components from Tripterygium wilfordii by macropo-rous resin. METHODS:The purification abilities of nine macroporous resins(ADS-5,ADS-8,HPD100,HPD300,HPD400, HPD450,HPD700,HPD722 and HPD750) were studied with the adsorption and desorption rates of triptolide,wilforlide,trip-tonide and tripterine as the index by static adsorption and desorption experiments for 4 active components from T. wilfordii,so as to screen optimal macroporous resins. Using transfer rate of active component as index,single factor test was used to investigate the effects of different sampling method,ratio of mixing sample to total resin quantity,ratio of resin to medicinal material,cleaning so-lution(type,amount and cleaning flow rate),diameter- height ratio of resins,eluant(volume,flow rate)on adsorption,so as to determine the optimal elution technology;validation test was also conducted. RESULTS:HPD722 macroporous resin was chosen as adsorption resin;ratio of diameter to height was 1∶10,resin-medicinal material ratio was 1∶2,and ratio of mixing sample to to-tal resin quantity was 1∶10;wet column installing and mixing resin for sample loading were adopted. The macroporous resin was washed with 12 BV 20%ethanol at the rate of 12 BV/h,and then eluted with 12 BV 95%ethanol at the rate of 6 BV/h. The verifi-cation test results showed that the total transfer rate of 4 active components from T. wilfordii was more than 90%(RSD=0.99%, n=3). CONCLUSIONS:The optimized technology is stable and feasible,and suitable for the purification of 4 active components from T. wilfordii.

Objective To evaluate the efficacy and safety of using alteplase for intravenous thrombolysis combined with vascular intervention for treatment of patients with acute intracranial large arterial occlusion.Methods Clinical data of 7 patients with acute intracranial large arterial occlusion treated by using alteplase for intravenous thrombolysis combined with intravascular intervention admitted to Department of Neurosurgery in Affiliated Hospital of the Logistics University of Cinese People's Armed Police Force from July 2015 to August 2016 were retrospectively analyzed.All the 7 patients were treated by alteplase dry powder (50 mg or 20 mg each ampule),solvent dose was 0.9 mg/kg,the maximum dose being ＜ 90 mg,firstly 10％ dose was intravenously injected,and the remaining dose was continuously infused into a vein in 60 minutes,during or after intravenous thrombolysis,digital subtraction angiography (DSA) was performed immediately,and according to the results of the angiography,at least one of the 3 kinds of intravascular mechanical intervention therapy,thrombectomy,balloon dilatation or stent placement,were chosen,and 24 hours after surgery,the anti-platelet aggregation drug and calcium channel antagonists were given,The effect of interventional therapy was analyzed,and the clinical outcome of 90-day treatment was evaluated.Results The mean age of the patients was (60.0 ± 12.6) years.Seven patients all successfully completed the treatment,and satisfactory re-canalization was achieved [they all obtained grade 3 or 2b in accord with the gradation of Thrombolysis in Cerebral Ischemia Scale (TICI)] in all the 7 cases (100％),after treatment,the National Institutes of Health Stroke Scale (NIHSS) score was significantly lower than that before treatment (5.86 ±4.10 vs.19.71 ± 5.56,P ＜0.01).Clinical outcome of 90-day follow up was excellent [Modified Rankin Scale (mRS) score 0-2] in 3 cases (42.8％).Conclusion Using alteplase for intravenous thrombolysis combined with endovascular intervention for treatment of patients with acute intracranial large arterial occlusion can achieve good re-canalization rate.