Antigens, CD20 ; metabolism ; Humans ; Ki-67 Antigen ; metabolism ; Liver ; pathology ; Liver Neoplasms ; immunology ; pathology ; Lymphoma, B-Cell ; immunology ; pathology ; Lymphoma, Large B-Cell, Diffuse ; immunology ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness

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.

Consensus Development Conferences as Topic ; Endoscopy ; Europe ; Humans ; Nose Neoplasms ; diagnosis ; surgery ; Skull Base Neoplasms ; diagnosis ; surgery

Adolescent ; Ethmoid Sinus ; Humans ; Male ; Osteoma ; complications ; Skull Neoplasms ; complications ; Vision, Low ; etiology

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 determine the 95 ％ effective target plasma concentration (EC95) of remifentanil for tracheal tube tolerance during the recovery period from anesthesia following cervical spine surgery.Methods Thirty ASA Ⅰ or Ⅱ patients,aged 18-60 yr,weighing 50-80 kg,scheduled for elective cervical spine surgery under total intravenous anesthesia,were enrolled in this study.Anesthesia was induced with iv injection of propofol,sufentanil and rocuronium.The patients were tracheal intubated and mechanically ventilated.Anesthesia was maintained with iv infusion of propofol and target-controlled infusion of remifentanil.The target plasma concentration (Cp) of remifentanil was set at 4-6 μg/L.BIS value was maintained at 40-60.Infusion of propofol was stopped at the end of surgery.Participants were allocated to a dose of remifentanil by 3-patient cohorts.Six Cps were selected from 1.0-3.5 μg/L before beginning and they were 1.0,1.5,2.0,2.5,3.0,3.5 μg/L.The Cp of remifentanil was 3.0 μg/L in the first cohort.After completion of the trial in each cohort,the posterior probability of each concentration was calculated according to the condition of sedation/analgesia and anterior probability of each concentration.The concentration with the posterior probability closest to 95 ％ was chosen as Cp in the next cohort.The concentration-probability curve was made according to the posterior probability of each concentration,and then EC95 and 95 ％ confidence interval of remifentanil were calculated.Results The EC95 and 95 ％ confidence interval of remifentanil were 2.77μg/L (2.65-2.83 μg/L) for tracheal tube tolerance during the recovery period from anesthesia following cervical spine surgery.Conclusion The EC95 of remifentanil for tracheal tube tolerance during the recovery period from anesthesia is 2.77 μg/L in patients undergoing cervical spine surgery.

Objective To explore the influencing factors of degenerative lumbar spondylolisthesis (DLS).Methods Sixty-one DLS patients (DLS group) were enrolled in this study.The control group was formed by 61 patients with same gender and age matched.The lesion position was L4 in two groups.The correlation of intervertebral joint angle,degeneration degree,lumbosacral angle and lumbar spondylolisthesis were analyzed.Results The intervertebral joint and sagittal angle in DLS group was less than that in control group[(37.11 ± 7.52)° vs.(42.44 ± 7.61)°] (P ＜ 0.01).The lumbosacral angle between two groups had no significant difference (P ＞ 0.05).The degeneration degree between two groups had significant difference (P ＜ 0.01).Conclusion Intervertebral joint and sagittal angle and intervertebral joint degeneration degree have important roles in DLS.

AIM: Using optical coherence tomography angiography (OCTA) to observe the changes and clinical significance of macular blood flow density in patients with diabetic retinopathy (DR).METHODS: Totally 47 eyes (28 patients) with diabetic retinopathy (DR) were enrolled in the DR group.According to the international clinical grading criteria of diabetic retinopathy, 30 eyes (19 patients) with non-proliferative diabetic retinopathy were classified as the NPDR group, and 17 eyes (11 patients) with proliferative diabetic retinopathy were classified as PDR group.A total of 46 (27 subjects) healthy eyes with matched age were enrolled in the control group.All the subjects underwent the 3mm×3mm scanning of macular retina by optical coherence tomography angiography (OCTA), obtaining 4 levels of macular blood flow density map.The macular blood flow density at 3 levels, including superficial retinal layer, deep retinal layer and choroidal capillaries layer, were measured.RESULTS: The macular blood flow density of superfical retinal layer, deep retinal layer and choroidal capillaries layer in DR group were 0.4963±0.0840, 0.4798±0.0801 and 0.5290±0.0528, respectively.Among them, the blood flow density of each layer were 0.5064±0.0843,0.4983±0.0766,0.5345±0.0529, respectively, for the NPDR group, and were 0.4786±0.0830, 0.4473±0.0778,0.5192±0.0526, respectively, for the PDR group.For the control group, the density of each layers were 0.5919±0.0704, 0.6301±0.0527, 0.5691±0.0169, respectively.The macular blood flow density was significantly different in the superficial retinal layer, deep retinal layer and choroidal capillary layer between the control group and the NPDR group, as well as the PDR group and the DR group (total P<0.001).Statistically significant difference was found between the NPDR group and the PDR group in the deep retina layer (P=0.029), but not in the superficial retina layer and choroid capillary layer (P=0.236, 0.268).CONCLUSION: Compared with the control group, the macular blood flow density of superficial retinal layer, deep retinal layer and choroidal capillary layer in the patients with diabetic retinopathy decreased significantly.It indicated that the macular ischemia existed in both retina and choroid.By quantitatively measurement of the macular blood flow, OCTA may be used for monitoring the progression of diabetes, and early detection of diabetic retinopathy.

Background In vitro study showed that chemotaxis consist of chemotaxis factor 4(CXCR4)and stromal cells derived factor-1(SDF-1)and may play a role in the orientation and migration of retinal progenitor cells (RPCs)toward lesion.Overexpression of CXCR4 in RPCs can enhance the chemotaxis activity.Objective This work was to explore the feasibility and underlying mechanism of up-regulation of CXCR4 on RPCs induced by hypoxia.Methods RPCs were retained in an incubator with normal O2volume(16％)or hypoxia condition(10％ O2)for 12 hours and 24 hours respectively.Flow cytometer cell analysis screening(FACS)was conduced to measure the proportion of CXCR4-expressing cells,and CXCR4,HIF-1 mRNA were analyzed by reverse transcription-polymerse chain reaction(RT-PCR).The chemotical effect of 30 mg/L SDF-1 to RPCs cultured under the hypoxia condition was assessed using Boyden chamber.Results The expression level of CXCR4(CXCR4 mRNA/β-actin mRNA)inRPCs cultured by 10％ O2 for 12 and 24 hours were 0.28+0.07and 0.48+0.17 and increased by 1.75 and 3.00 fold more than that of 16％ O2 culture group(0.16+0.02)(P＜0.01).The expression level of HIF-1 mRNA(HIF-1 mRNA/β-actin mRNA)in RPCs cultured by 10％ O2 for 12 and 24 hours were 0.18 ±0.07and 0.38 ±0.13 and increased by 3.00 and 6.30 fold more than that of 16％ O2 culture group(0.06±0.01)(P＜0.01).The chemotical effect of 30 μg/L SDF-1 to RPCs increased from 13.00％ in 16％ O2 culture group to 36.00％ and 46.00％ in the cells cultured by 10％ O2for 12 and 24 hours.FACS revealed that the proportion of CXCR4+ cells in hypoxia-exposure for 12 and 24 hours were 26.90％ and 46.10％,respectively,but that in 16％ O2 culture group was 9.10％,showing a statistically significant difference(P ＜ 0.01).Conclusions RPCs induced by hypoxia can enhance the expression of CXCR4 in RPE cells and the chemotaxia to SDF-1.The overexpression of H1F-1 in RPCs may be involved in the up-regulation of CXCR4 expression.

OBJECTIVETo detect the expression of PSF1 (partner of Sld five 1) in colon cancer specimens, and to explore the effect of RNA interference targeting PSF1 on the proliferation of colon cancer cells and its mechanism.

METHODSExpression level of PSF1 protein in colon cancer specimens was detected by Western blot in 40 patients with colon cancer from May 2004 to December 2006. The short hairpin RNA (shRNA) plasmid targeting PSF1 was transfected into LOVO, HT-29 and HCT116 cells with liposome, then the expression level of PSF1 protein was measured by Western blot, the effect of PSF1 shRNA plasmid transfection on cell proliferation by MTT assay, anchorage-independent growth by soft agar colomy-formation assay, and PSF2, PSF3 and SLD5 mRNA expression by quantitative reverse transcription polymerase chain reaction.

RESULTSThe relative expression level of PSF1 protein in colon cancer tissues was 0.485±0.113, which was significantly higher than that in adjacent normal mucosa tissues (0.056±0.014, P<0.01). Western blot showed that the expression level of PSF1 protein was significantly decreased in colon cancer cells transfected with PSF1 shRNA plasmid. After PSF1 shRNA plasmid transfection, cell proliferation was significantly suppressed, the soft agar colony-forming rates of LOVO, HT-29 and HCT116 cells were significantly lower than those in control groups (P<0.05), meanwhile the expression levels of PSF2, PSF3 and SLD5 mRNA were significantly decreased (P<0.05).

CONCLUSIONSPSF1 is significantly up-regulated in colon cancer tissues compared with adjacent normal mucosa tissues. ShRNA plasmid targeting PSF1 can inhibit the expression of PSF1 gene, suppress the proliferation of colon cancer cells, suggesting that it may be a new therapeutic target for colon cancer.

ATP-Binding Cassette Sub-Family B Member 2 ; ATP-Binding Cassette Transporters ; genetics ; metabolism ; Adult ; Aged ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Middle Aged ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection