0.05);the TAV of the drug plus radiation dose of 20,25Gy groups were much smaller than that of the radiation alone groups (P

Objective To explore the expression of methyl CpG binding protein 2 (MeCP2) in gastric cancer tissues and its role in multi-drug resistance (MDR) in gastric cancer cells.Methods The expression of MeCP2 in 90 gastric cancer tissues and the adjacent normal tissues was detected by immunohistochemistry,and the relationship between MeCP2 expression level and clinicopathological parameters was analyzed.The expression level of MeCP2 in gastric cancer cell line SGC7901,MDR cell variants SGC7901/doxorubicin hydrochloride(ADR) and SGC7901/vincristine(VCR)was determined by Western blot.After the expression of MeCP2 was silenced by short interference RNA (siRNA),half inhibitory concentration (IC50) of 5-fluorouraeil and cisplatin in gastric cancer cell was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT) assay.The t test or chi square test was performed for statistical analysis.Results The positive rate of MeCP2 expression in gastric cancer tissues was 72.2％ (65/90),which was significantly lower than that in adjacent normal tissues (93.3％,84/ 90),and the difference was statistically significant (x2 =14.068,P＜0.01).MeCP2 expression was not correlated with age,tumor maximum diameter and lymph node metastasis (all P＞0.05),however which was related to gender,clinical TNM stages and distant metastasis (x2=4.680,4.186 and 4.327;aH P＜ 0.05).The gray scale ratios of MeCP2/β-actin in SGC7901/ADR and SGC7901/VCR were 0.593 7 ± 0.030 5 and 0.651 2 ± 0.018 6,which were lower than that of parental SGC7901 cells (1),and the differences were statistically significant (t =23.080,17.360;both P ＜ 0.01).After the expression of MeCP2 was silenced by siRNA,the IC50 of 5-fluorouracil and cisplatin in SGC7901 transfected with MeCP2 specific siRNA were (11.540 0±0.469 3) μg/mL and (2.273 0±0.265 4) μg/mL,which were higher than the IC50 of 5-fluorouracil and cisplatin in SGC7901 transfected with non-related oligonucleotide sequence ((8.663 0±0.160 1) μg/mL and (0.884 0 ±0.038 6) μg/mL),respectively.The differences were statistically significant (t =15.380 and 8.153;both P ＜ 0.01).Conclusions The expression of MeCP2 in gastric cancer tissues significantly decreased,which was correlated with clinical stages,distant metastasis.MeCP2 can inhibit the MDR of gastric cancer cell,which indicated the dysregulated expression of MeCP2 might participate in the genesis and development of gastric cancer.

Objective To investigate the changes of learning and memory function ,vascular endothelial growth factor (VEGF) and heme oxygenase‐1(HO‐1) expression in cerebral cortex and hippocampus of chronic ischemic vascular dementia rats . Methods Thirty‐six healthy SD rats were divided into the control group ,sham operation group and model group ,12 cases in each group .The chronic ischemic vascular dementia rat model was established by the permanent bilateral carotid artery occlusion The sham operation group received the same treatment to the model group except without bilateral carotid artery occlusion .The learning and memory abilities were tested by the Morris water maze experiment and climbing rope strength experiment at 1 ,4 ,8 ,12 weeks respectively .The expressions of VEGF and HO‐1 in rat cerebral cortex and hippocampus was determined by immunohistochemical SP technique .Results The escape latency time at 8 ,12 weeks in the model group was longer than that in the sham operation group and control group ,and the number of crossing the platform was less than that in the sham operation group and control group ,the differences were statistically significant (P<0 .05);the time of climbing at 1 -12 weeks had no statistical difference between the model group and the sham operation group and between the model group and the control group (P>0 .05) .The positive expression of HO‐1 and VEGF protein contents in the control group and sham operation group was less than that in the model group with sta‐tistical difference(P<0 .05) .Conclusion Chronic cerebral hypoperfusion has a permanent damage to the learning and memory abil‐ities in rats ,while has no influence on the motor function .VEGF and HO‐1 may play a protective role in chronic cerebral ischemia .

ObjectiveTo investigate the effects of heterogeneous phosphatase and tensinhomologue deleted on chromosome ten (PTEN) on cell cycles,proliferation,invasion,tumorigenicity,metastasis and the expressions of vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR)proteins in human pancreas cancer cell line ( ASPC-1 ).MethodsASPC-1 cells was transfected with plasmid pE-PTEN containing PTEN,and empty plasmid pE-PTEN transfection was used as control,then ASPC-1-pE-PTEN (A-pE-P) cell and ASPC-1-pE (A-pE) cell was obtained.The expression of PTEN mRNA was determined by RT-PCR. PTEN,VEGF and EGFR proteins were measured by cell immunohistochemical method.Clone formation assay was used to observe the numbers of clone.Transwell was used to test the invasion ability of cells.The growths of tumor were detected by nude mice subcutaneous injection of cancer cells in vivo.Results Compared with ASPC-1,the expressions of PTEN mRNA of A-pE-P increased by 179.3％,and the expressions of PTEN protein were also significantly increased.The expressions of VEGF protein were significantly decreased.The expressions of EGFR protein were not significantly changed.Number of G2/M phase cells was significantly increased from (26.81 ± 1.03)％ to (31.5 ± 1.76)％ (P ＜0.05).The numbers of clone was decreased by 28％ (P ＜0.05).The number of penetrating cells was decreased[(46.3 ±6.6) vs (63.8 ±7.5) per high power field,P ＜0.05].The tumor volumes were significantly reduced [(142.4 ±30.9) vs (202.7 ±43.6) mm3,P ＜0.05].The tumor inhibitory rate was 42.4％.The distant metastases were significantly reduced [(2.0 ±0.7) vs (5.0 ± 1.3),P ＜0.01 ].Conclusions Heterogeneous PTEN can not only inhibit the proliferation,invasion and metantasis of ASPC-1 cells,arrest the cell growth at G2/M phase,but also decrease the expressions of VEGF.

Objective To investigate the mechanisms and effects of Sorafenib on cytotoxic sensitivity of allo-reactive natural killer(Allo-NK) cells against human multi-drug resistant nasopharyngeal carcinoma CNE2/DDP cells which expressing highly ATP-binding cassette superfamily G member 2(ABCG2)(abbr.as ABCG2HighCNE2/DDP cells).Methods ABCG2HighCNE2/DDP and Allo-NK cells were isolated by magnetic bead technique.The target cells were divided into 3 groups: a) treated group(ABCG2HighCNE2/DDP cells incubated with 10 ng/ml sorafenib for 4h);b) untreated group(conventionally cultured ABCG2HighCNE2/DDP cells);and c) control group(conventionally cultured K562 cells).Expression rates of ABCG2 in treated and untreated groups,and of five NKG2D-ligands(MICA,MICB,ULBP1,ULBP2,ULBP3) were evaluated by flow cytometry.The cytotoxic effects of NK cells against different groups of target cells were detected with LDH releasing assay.Results Expression rate of ABCG2 in isolated CNE2/DDP cells was 91.40%?2.32%.The purity of sorted CD3-CD16+CD56+ Allo-NK cells was 90% and higher.The expression rates of NKG2D-ligands(MICA,MICB,ULBP1,ULBP2 and ULBP3) in untreated group were 2.92%?0.33%,4.27%?0.33%,5.80%?0.62%,11.10%?3.15% and 7.75%?1.14%,respectively,which were remarkablely higher than that in treated group(10.38%?1.23%,10.68%?1.26%,11.62%?1.22%,43.24%?4.42% and 11.91%?0.88%,respectively,P

Objective:To identify differentially expressed saliva proteins of oral lichen planus(OLP)patients by two-dimensional fluo-rescence difference electrophoresis(2-D DIGE)and mass spectrometry(MS).Methods:3 pairs of saliva samples from OLP patients and matched healthy adults were collected.Saliva proteins were separated by 2-D DIGE and identified by liquid chromatography-mass spectrometry(LC-MS).Results:SDS-PAGE examination showed that the electrophoresis bands were clear and protein loss was rare. Protein dots were highly reproducible by 2-D DIGE.In average,the abundance of (31 7 ±71 )saliva protein spots were found in OLP pa-tients.4 highly reproducible spots were identified to be secretory IgA1 ,zincα-2-glycoprotein,salivary amylase and serum albumin by LC-MS and they were at higher level in OLP patients than those in the healthy controls.Conclusion:Secretory IgA1 ,zincα-2-glyco-protein,salivary amylase and serum albumin are highly expressed in the saliva of OLP patients,and may be related to the occurrence and development of oral lichen planus.

Objective To investigate the effects of PTEN and KAI1 double gene transfection on proliferation,metastasis in AsPC1 pancreatic cancer cells under hypoxic condition.MethodsRecombinant vectors that over expressing PTEN and KAI1 protein which was established previously were double transfected into hypoxic AsPC1 cells.Western blot was performed to measure the expression level of PTEN and KAI1.Then,cell proliferation was detected by MTT,and colony forming assay were used to test the ability of tumor cells forming colonies,and transwell assay was used to evaluate metastatic function.ResultsAfter double gene trnsfection,both PTEN and KAI1 protein expression was significantly up-regulated,which was 2.05 and 1.5 1 folds higher than that of empty vector group; and cell proliferation of hypoxic AsPC1 cells was suppressed significantly (0.5 vs 0.8,P =0.00031 ),number of colony formation was significantly decreased [ (41.67 ± 5.03) vs (86.00 ± 7.81 ),P =0.017) ] ; the capacity of metastasis was significantly decreased (0.68 ± 0.05 vs 1.23 ± 0.03,P =0.0025 ).ConclusionsDouble gene transfection of PTEN and KAI1 could inhibit proliferation and metastatic activity of hypoxic AsPC1 cells,which might indicate that combined gene therapy may play a role in the treatment of pancreatic cancer.

Construction of the network course is one of the hot spots of teaching reform. With medical preparations for military struggle and the continuous improvement of dealing with unexpected natural disasters,the department of ophthalmology analyzed the characteristics,status and the advantages of network teaching of the"eye injury"carefully,to proceed with network cur-riculum reform. Teaching is designed through specific goals,processes,content,organization and development environment,and the other aspects.

Objective To observe the influence of renal sympathetic denervation (RSD) on renal interstitial fibrosis and transforming growth factor beta 1(TGF-β1) and microRNA-21 (miR-21) in rats with unilateral ureteral obstruction(UUO).Methods 40 male Wistar rats were randomly divided into UUO group (A group,n=10),sham UUO group (B group,n=10),RSD+UUO group (C group,n=1O) and RSD + sham UUO group (D group,n=10).Rats in A group and C group underwent unilateral ureteral ligation,while those in B group and D group underwent sham operation.Rats in C group and D group were followed by RSD.Rats were sacrificed at 21 days after the operation to evaluate the fibrosis by Masson staining.Immunohistochemical staining and Western blotting were used to detect the expressions of collagen I (COL-Ⅰ),collagen Ⅲ (COL-Ⅲ) and TGF-β1 in four groups.The expression of miR-21 was detected by fluorescence in situ hybridization (FISH) and quantitative real-time PCR (RT-qPCR).Results A large amount of collagen deposition was observed in the renal interstitial area in A and C group compared to either B or D group (P ＜ 0.05),but the change in C group was decreased significantly than that in A group (P ＜ 0.05).Similarly,the expressions of COL-Ⅰ,COL-Ⅲ,TGF-β1and miR-21 were obviously higher in A and C group compared to either B or D group (P ＜ 0.05),but those change in C group were decreased significantly than those in A group (P ＜ 0.05).The above indexes were not significantly different between B group and D group (P ＞ 0.05).Conclusion RSD may relieve the renal interstitial fibrosis in UUO rats,and down-regulate the expression of TGF-β1 and miR-21.

Objective: To observe the clinical effect of pestle needle combined with Chinese herbal fumigation on cervical spondylosis and provide a safe effective therapy for this condition. Methods: A total of 54 cases were randomly allocated into two groups (27 cases in each group) according to their sequence of consultation. Patients in both groups were treated with the same Chinese herbal fumigation. Patients in the treatment group were additionally treated with pestle needle therapy on a unique set ofBa Zhen points around Dazhui (GV 14), Fengfu (GV 16) and the distance between Naohu (GV 17) and Dazhui (GV 14) along theHe Chepathway, whereas patients in the control group were additionally treated with routine acupuncture therapy. Then the short-term and long-term efficacies were observed and compared after treatment using the visual analog scale (VAS) and pain rating index (PRI). Results: At the end of treatment, VAS scores were significantly decreased in both groups, and the VAS score in the treatment group was lower than that in the control group (P<0.05). The intra-group differences were statistically significant in VAS scores 1 month, 3 and 6 months after treatment (allP<0.05). At the end of treatment, the sensory and total PRI scores in the treatment group were significantly lower than those in the control group, showing statistically significant differences (both P<0.01); and there was no significant between-group difference (P>0.05) in the affective PRI score. At the end of treatment, the total effective rate was 85.2% in the treatment group, versus 65.4% in the control group, showing a statistical significance (P<0.05). The follow-up six months later showed that the total effective rate was 92.6% in the treatment group, versus 76.9% in the control group, showing a statistical significance (P<0.05). Conclusion: Pestle needle therapy is a stable and positive therapy for cervical spondylosis.