Objective To analyze the differentially expressed genes in esophageal squamons cell carcinoma (ESCC), para-cancerous tissue (PCT) and normal esophagus tissue (NET) using oligomicroarray and to identify the target genes related to the development and progression of esophageal carcinoma. Methods The total RNAs isolated from ESCC, PCT or NET using one step Trizol method were purified and reversely transcribed into cRNAs. The cRNAs were then fluorescence labeled and hybridized with Agilent oligomicroarray (21 074 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. The selected candidate genes were confirmed by real time real time fluorescent quantitative RT-PCR immunohistochemistry andWestern blotting.Results ① The oligomicroarray demonstrated that there were 38 up-regulated genes and 61 down-regulated genes. ② The real time fluorescent quantitative RT-PCR revealed that five genes (CTHRC1, INHBA, SPP1 ,LUM, HRK)were more differentially expressed in up-regulated genes. Of which, CTHRC1 displayed more disparity.③ Immunohistochemistry examination showed that the higher expression of CTHRC1 (56.5 %, 26/46) was observed in ESCC. There was significantly difference in expression of CTHRC1 between patients with or without lymph node metastasis (P<0.05). ④ CTHRC1 protein was expressed in both TE-13 and Eca-109 cell lines. Conclusion CTHRC1 is probably one of the most significant biomolecules in ESCC.

Objective To investigate the expression of Ezrin in gastric cancer tissues and its clinical significance.Methods Gastric cancer tissues and adjacent normal gastric tissues from 60 patients with gastric cancer were collected from June 2008 to May 2009 at the Southwest Hospital.The mRNA and protein expressions of Ezrin were detected by using the reverse transcription polymerase chain reaction and western blot.The relationship between Ezrin and the gender and age of patients,and tumor differentiation,pathological staging,depth of invasion and lymph node metastasis was analyzed.All data were analyzed using the t test,chi-square test and Spearman rank correlation.Results The Ezrin mRNA expression level was increased in 33 (55％) cases of adjacent normal gastric tissues and 21 (35％) cases of gastric cancer tissues; the Ezrin protein expression level was increased in 45 (75％) cases of adjacent normal gastric tissues and 22 (37％) cases of gastric cancer tissues.The mRNA and protein expressions of Ezrin in the normal adjacent gastric tissues were 1.30 ± 0.04 and 3.57 ± 0.45,respectively,which were significantly higher than 0.53 ± 0.36 and 0.96 ± 0.18 in the gastric cancer tissues ( t =5.309,22.617,P ＜ 0.05 ).The mRNA expression of Ezrin was positively correlated with the protein expression of Ezrin (r =0.602,P ＜ 0.05 ).The mRNA and protein expressions of Ezrin were related to the pathological stages,depth of invasion and state of lymph node metastasis (x2 =6.41,6.49,4.62; 5.40,8.87,4.12,P ＜ 0.05),but not to the gender,age and tumor differentiation (x2 =0.50,0.07,1.07 ; 0.01,1.16,1.96,P ＞ 0.05).Conclusion The mRNA and protein expressions of Ezrin are significantly decreased in the gastric cancer tissue,which might be responsible for genesis,development and metastasis of gastric cancer.

Objective To investigate the influence of gene therapy using survivin as a gene target on biological behavior of pancreatic carcinoma cell line. Methods Chemically synthesized siRNA and shRNA in pGCSi vector were used to silence survivin expression of pancreatic carcinoma cell line PaTu8988. The therapeutical effects of survivin as a gene target were evaluated through determination of the down-regulation of survivin gene expression, cellular shape, cell apoptosis, cell viability and apoptosis signal pathway changes. Results After transfection of different arrays of siRNA and shRNA vectors to silence the survivin expression, survivin mRNA and protein levels were significantly decreased (P < 0.05) ; PI staining revealed the presence of karyopyknosis, the cell apoptosis index was more than 20%; hypodiploid DNA content before G0/G1 detected by flow cytometry ; cell viability measured by MTT assay was significantly decreased (P <0.05) ; the activity of caspase-3 remarkably increased (P < 0. 05). Conclusions The pancreatic carcinoma cell line PaTu8988 be induced to promote spontaneous apoptosis procedure through silencing survivin expression by RNAi, which could accelerate carcinoma cell apoptosis and improve therapeutic effect on pancreatic carcinoma.

Objective To investigate the effects of hypoxia-inducible factor (HIF)-1α on glycolysis of human esophageal squamous carcinoma cells and the possible mechanism.Methods TE13 and Eca109 cells were cultured under normal oxygen (20％O2) and hypoxia (1％O2) conditions.The hypoxia was duration 6 hours,12 hours,24 hours and 48 hours.HIF-1α gene was stable silented by RNA interference method and TE13/small interfering HIF cells and Eca109/siHIF cells were obtained.The cell culture condition and time was same as TE13 and Eca109 cells.The changes of HIF-1α expression were detected by Western-blot.The changes of lactic acid concentration in cell culture supernatant were determined by Spectrophotometry.The changes of glucose transporter-1 (GLUT-1) and lactic dehydrogenase A (LDHA) expression at mRNA level were examined by realtime polymerase chain reaction.The changes of GLUT-1 and LDHA expression at protein level were tested by Western blot.Using t or t' test to analyze the effects of hypoxia duration on HIF-1αexpression at protein level.One-way ANOVA was applied for the difference analysis between the groups.Results In TE13 and Eca109 cells,the HIF-1α expression significantly increased under hypoxia condition and reached the peak at 12 hour (t=6.11,8.31; both P＜0.05).The lactic acid secretion of TE13/siHIF cells and Eca109/siHIF cells was (1.24±0.33) and (1.28±0.37) mmol/L,which significantly decreased when compared with TE13 and Eca109 cells [(3.25±1.34) and (4.91±1.69) mmol/L,t=2.53,3.59,both P＜0.05].The lactic acid secretion of TE13 and Eca109 cells significantly increased after hypoxia [(6.48±1.73) and (8.02± 1.95) mmol/L,t=2.715,2.050,both P＜0.05].There was no significant lactic acid secretion in TE13/siHIF cells and Eca109/siHIF cells after hypoxia (P ＞ 0.05).The expressions of GLUT-1 and LDHA at mRNA level were significantly suppressed in TE13/siHIF cells and Eca109/siHIF cells (normal oxygen:t=6.98,3.92,7.25,3.67,all P＜0.05).The expression of GLUT-1 at protein level remarkably weaked (normal oxygen:t=4.57、16.56,hypoxia:t=6.19、6.09,all P＜0.05),while the expression of LDHA at protein level slightly decreased (P＞0.05).Conclusions The level of glycolysis can be lowered by suppression HIF-1α expression in human esophageal squamous carcinoma cells.The pathway may be involved in the suppression of GLUT-1 and LDHA expression.Except for HIF-1α,there may be other regulating factors in LDHA protein expression at same time.

Adult ; Female ; Humans ; Male ; Mercury Poisoning ; diagnosis ; therapy ; Middle Aged

Humans ; Male ; Middle Aged ; Silicosis ; surgery

Objective To investigate the effect of intra-articular carboxymethylated ehitosan(CM- CTS)injection on inducible nitric oxide synthase(iNOS)expression in cartilage at the early stage of os- teoarthfitis(OA).Methods Thirty-two white rabbits were underwent unilateral anterior cruciate ligament transection(ACLT)and were randomly divided into 4 groups 5 weeks after transection.Rabbits of group A re- ceived 0.3 ml of 2% high molecular weight CMCTS(H-CMCTS)once every two weeks.Rabbits in group B were treated using 2% low molecular weight(L-CMCTS)CMCTS at:the same intervals.Group C rabbits were injected intra-articularly with 0.3 ml of 1% sodium hyaluronate(Na-HA)once a week.Animals of group D were not injected.At sacrifice,11 weeks following surgery,the expression of iNOS in cartilages was analyzed by immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR)methods.Results Both immunohistochemistry and RT-PCR showed that the level of iNOS expression of cartilage in CMCTS in- jection groups was lower than that in Na-HA injection group and the untreated group.There was no significant difference in iNOS expression between the two different molecular weight CMCTS injection groups. No signifi- cant difference of iNOS expression in cartilage was found between Na-HA injection group and the untreated group.Conclusion CMCTS suppresses iNOS expression in cartilage during the early stage of OA.Na-HA treatment has no effect on iNOS expression in cartilage.

Objective To observe the primary clinical results of Gore （R） Viabahn endoprosthesis for atherosclerotic stenosis or occlusion in superficial femoral artery (SFA).Methods From March 2013 to November 2013,45 consecutive patients (54 limbs) who had ischemic symptom due to stenosis or occlusion of superficial femoral artery were treated by endovascular deployment of Gore （R） Viabahn endoprosthesis.We observed patency rate,improvement of Rutherford classification and ankle-brachial index,limb salvage rate and survival rate after 1,3 and 6 months.Results 43 patients (52 limbs)were followed-up,among which 32 patients(38 limbs)belonged to TASC Ⅱ type C or type D lesion.The mean (± SD) length of treated segment in 52 limbs was(19 ±4)cm.The rate of postoperative patency on duplex ultrasonography:one month,3 months and 6 months were 96.15％ (50 limbs),92.31％ (48 limbs)and 90.38％ (47 limbs) ; Ankle-brachial index increased from(0.32 ±0.20)to(0.68 ±0.18) (t =3.180,P ＜ 0.005) after 6 months ; at 6 months limb salvage rate was 96.08 percent (49 limbs),and survival rate was 97.67％ (42 patients).Conclusions Implantation of Gore （R） Viabahn endoprosthesis in atherosclerotic stenotic or occluded superficial femoral artery safely achieves a definite primary clinical effectiveness.

Objective To summarize the experience in application of da Vinci robotic surgical system in radical resection of rectal cancer,and investigate the proper position of trocars and operative techniques.Methods The clinical data of 13 patients who received radical resection of rectal cancer accomplished by the da Vinci robotic surgical system at the Southwest Hospital from February 2010 to February 2011 were retrospectively analyzed.The patients were in lithotomy position and received combined intravenous anesthesia.Five or 4 trocars were used.Miles procedures were performed on patients with lower tumor position,and the other patients received Dixon procedure.Results The operation was successfully performed on all patients.Five trocars were selected for the first 3 patients,and 4 trocars for the other 10 patients.Nine Dixon procedures and 4 Miles procedures were selected.The mean operation time was 217.3 minutes (range,160-260 minutes).The mean operative blood loss was 53.3 ml (range,40-70 ml) in Dixon procedure and 120.0 ml (range,90-130 ml) in the Miles procedure,and no blood transfusion was needed.The mean number of lymph nodes dissected was 13.9 (range,8-21 ),and the time to bowel movement was 3.2 days (range,2-5 days).Two patients were complicated with pulmonary infection,1 with urinary tract infection,and they were cured by antimicrobial therapy.No other morbidity or mortality was found.The results of postoperative pathological examination showed that there were no residual cancer cells at the resection margin,and the distance between the resection margin and the tumor was 6.3 cm (range,3-10 cm).There were 1 patient in stage Ⅰ,5 in stage Ⅱ and 7 in stage Ⅲ.The mean time of follow-up was 5.9 months (range,3-12 months),and no recurrence or metastasis was found during follow-up.ConclusionsRadical resection of rectal cancer with da Vinci robotic surgical system utilizing 4 trocars has the advantages of minimally invasive surgery with fast recovery as well as the ease of dissection afforded by the surgical robot.

ObjectiveTo evaluate the feasibility, safety and the long-termoutcomes of laparoscopy-assisted distal gastrectomy (LADG) for advanced gastric cancer (AGC).MethodsWe retrospectively analyzed the clinical and follow-up data of 346 cases after LADG from January 2004 to June 2009, compared with 313 cases after conventional open distal gastrectomy (ODG) for advanced gastric cancer at the same period at our hospital. The surgical safety, postoperative complications, survival rate, and the recurrence and metastasis of cancer were compared.ResultsThere was no significant difference at the average time of LADG and ODG procedures (211 ± 56) min vs.(204 ±41 ) min, but blood loss during operation and length of incision in LADG group were significantly less than in the ODG group. The proximal and distal length were, respectively, (6. 3 ± 2. 0) cm and (5. 7 ± 1.7 ) cm in LADG group and (6. 3 ±2. 1 ) cm and (5.6 ± 1.6) cm in ODG group, the difference was not significant. The number of lymph node dissections was also similar: (33 ± 13) in LADG group and (33 ± 16) in ODG group. The incidence of postoperative complications in LADG group was significantly lower than that in ODG group ( 6. 7％ vs.13. 1％, P ＜ 0. 05). During the follow-up period of 6-72 months (average 37 months), the 1-, 3-and 5-year survival rates were, respectively, 87. 2％, 57. 2％ and 50. 3％ in LADG group and 87. 1％, 54. 1％and 49. 2％ in ODG group, the difference was not significant. The differences in recurrence and metastasis between the two groups were not statistically significant.ConclsionLaparoscopy-assisted gastrectomy for advanced gastric cancer is not significantly different with open surgery in postoperative survival rate or recurrence. It is less traumatic and of fewer complications.