Objective:To analyze the clinical manifestations and esophageal motility characteristics of patients with gastroesophageal reflux disease (GERD) and extra-esophageal symptoms.Methods:From January 1 to October 30, 2018, at PLA Rocket Force Characteristic Medical Center, 180 hospitalized patients diagnosed with GERD and extra-esophageal symptoms were retrospectively analyzed. The patients were divided into laryngopharyngeal symptom group (65 cases), airway symptom group (58 cases) and mixed symptom group (57 cases). General data, clinical symptoms, gastroscopic manitestations, the results of high-resolution esophageal manometry and 24-hour multichannel intraluminal impedance and pH monitoring of each group were analyzed and compared. Chi-square test and one-way analysis of variance were used for statistical analysis.Results:The patients aged <40, 40 to 60 and >60 years accounted for 12.8% (23/180), 53.3% (96/180) and 33.9% (61/180), respectively, and the difference was statistically significant ( χ2=12.030, P=0.017). There were 18.9%(34/180) of patients without typical reflux symptoms. There were statistically significant differences in the incidence of ectopic esophagogastric mucosa or Barrett esophagus under gastroscopy between laryngopharyngeal symptom group, airway symptom group and mixed symptom group (21.5%, 14/65; 5.2%, 3/58 and 8.8%, 5/57, respectively) ( χ2=8.578, P=0.014). There were no statistically significant differences in the lower esophageal sphincter pressure (LESP), upper esophageal sphincter pressure or distal contractile integral between laryngopharyngeal symptom group, airway symptom group and mixed symptom group ((8.57±0.76), (8.87±0.79), and (10.51±0.97) mmHg (1 mmHg=0.133 kPa); (44.75±2.86), (42.81±4.06), and (39.14±3.20) mmHg; (506.13±64.30), (432.59±78.10), and (682.99±82.28) mmHg·s·cm)(all P>0.05). The DeMeester score of laryngopharyngeal symptom group , mixed symptom group and airway symptom group was (14.33±2.09), (21.94±5.30) and (30.47±5.85) points, respectively, and the difference was statistically significant ( F=3.226, P=0.043). The results of multi-channel impedance monitoring showed that acid reflux and weak acid reflux were the main reflux in the patients, which accounted for 55.5% (76/137) and 34.3% (47/137), respectively. Among 87.6% (120/137) of the patients, reflux mainly occurred in the upright position while 12.4% (17/137) of the patients had reflux in the supine position. Conclusions:The extra-esophageal symptoms of GERD is associated with age. Ectopic esophagogastric mucosa or Barrett esophagus are more common in GERD patients with laryngopharyngeal symptoms. There are more acid exposure and pathologic acid reflux in GERD mainly with airway symptoms. Weak acid reflux at upright position plays an important role in the reflux mechanism of GERD with extra-esophageal symptoms.

Objective To explore the relationship between the expression of thrombospondin-2 (TSP-2), the changes of MVD and angiogenesis in non-small cell lung cancer (NSCLC). Methods By using SP immunohistochemical staining, the expression of TSP-2 and microvessel counting were evaluated in surgically resected specimens from 55 patients with NSCLC and 30 patients' pancancerous tissues, using the anti-TSP-2 and anti-CD34 monoclonal antibodies respectively. Results Active angiogenesis took place in NSCLC tumor tissues. Microvessel density (MVD) was associated with metastasis and clinical stage of NSCLC. Expression level of TSP-2 in the tumor tissues of 55 NSCLC patients was significantly higher than that in the pancancerous tissues from other 30 patients (149.10?2.94 vs. 145.70?4.74, P

Objective To observe the effect of RNAi that silences MTA1 gene on invasion and migration of esophageal carcinoma 9706 cells. Methods The siRNA expression vector that silences MTA1 gene was transfected into EC9706 cells by liposome. MTA1 mRNA and protein expressions were detected through quantitative RT-PCR and Western blot, respectively. The invasion and migration of EC9706 cells were evaluated by scrape wound healing assay and cell invasion assay in vitro. Results MTA1 gene expression significantly decreased. The scrape wound of EC9706 cells healed more slowly and the cell population that cut through Matrigel were less in the EC9706 cells transfected with siRNA expression vector than non-transfected EC9706 cells and the EC9706 cells transfected with blank vector (P

ObjectiveTo evaluate the anticancer activity of 5'-dexoxy-fluorouridine on colon cancer experimental models in BALB/C mice,compared with 5'-fluorouracil,an anticancer agent widely used in clinic,meanwhile,examined the conversion of 5'-Dexoxy-fluorouridine to 5' -fluorouracil in cancer tissues and serum of mouse models.MethodsThe xenografts of mouse colon cancer cell line CT 26 were transplantated to cecum in 60 male BALB/C mice.Three days lated,these mice were divided into 3 groups and intro- peritoneally injected:( 1 ) 5' - dexoxy- fluorouridine 0.1 mg/g,(2) 5' - Fluorouracil 0.02 mg/g,(3)0.9％ sodium chloride 0.4 mL (as a control),respectively.Two and three weeks later,6 mice were sacririced in every group respectively to measure the weight of tumors and bodies,to examine the Hb,RBC,WBC,PLT,AST,ALT,UREA,and CREA in blood.The rest 8 mice in each group were fed generally,and the survival time from operation to natural death was recorded.In addition,14 mice with xenografts of CT 26 about 2 weeks,were divided into 2 groups averagely,5' -dexoxy-fluorouridine 0.1 mg/g and 5' -fluorouracil 0.02 mg/g were intro-peritoneally injected respectively.Fifteen min later,the converted 5' -fluorouracil was detected from the blood and tumor tissues in sacrificed mice.ResultsThe lest tumor average weight was found in the mice injected 5 '-dexoxy-fluorouridine,being (0.07 ± 0.12) g and (0.24g ±0.29) g for the mice sacrificed at 2 and 3 weeks later,respectively.The average survival time for rest mice was ( 32.6 ± 8.9) d.The average tumor weight in 5' - fluorouracil group was (0.74 ± 0.43 ) g and ( 1.13 ±0.75) g at 2 and 3 weeks later,and the average survival time for the rest was (22.8 ±5.9)d,respectively.The average tumor weight in the control group was (0.70 ±0.47) g and ( 1.93 ±0.83) g at 2 and 3 weeks,and the average survival time for the rest was ( 17.5 ± 2.8 ) d.Either the average tumor weight or average survival time in the mice of 5 ' -dexoxy-fluorouridine group was significantly differen from either 5' -fluorouracil group or control (P ＜ 0.05 ).However,there was no significant difference for the numbers of WBC,PLC,Hb,and some function examination of liver and kidney among 3 group mice,besides the loss of weights in 5'-fluorouracil group mice after operation and medicine therapy which was significantly obvious than that in 5' -deoxy-fluorouridine and control groups ( P ＜ 0.05 ).In addition,( 54.71 ± 12.82) μg/g 5' -fluorouracil was detected in xenografts of mice injected 5' -dexoxy-fluorouridine 15 min later,which was the 6.20 folds of 5' -fluorouracil detected in serum from sthe ame group,P ＜0.05.However,( 133.35 ±20.69) μg/m 5'-fluorouracil were detected in serum of mice after 5' -fluorouracil were injected 15 min later,which was the 1.55 folds of 5' -fluorouracil detected in the xenografts from same group ( P ＜ 0.05 ).ConclusionsIn colon cancer tissues of mouse experimental models,5' - dexoxy- fluorouridine could be converted effectively to 5'-fluorouracil,an obvious high concentration being detected in serum of mice than in cancer tissues.The anticancer effect of 5'-dexoxy-fluorouridine on mouse colon cancer models was more effective than 5'-fluorouracil,resulting in a longer survival duration,less side effect and no significant injury on liver and kidney functions.However,the mechanism of 5' -dexoxy-fluorouridine converted to 5' -fluorouracil in cancer tissue is needed further investigation.

Objective To study the effects of Alternariol(AOH) on DNA polymerase ?(DNA POL?)expression in NIH3T3 cells.Methods RT-PCR,Immunocytochemistry and Western blot were used to detected mRNA and the protein levels of DNA POL? in NIH3T3 cell line induced by AOH.Results The expression of DNA POL? in NIH3T3 cells contaminated by AOH was significantly higher than that in the control group(P

Objective To discuss the feasibility on building lewis lung carcinoma mouse models through different methods and improve the methods. Methods The method of culture LLC cells in vitro, trypsin digestion method, Ⅳ collagenase method and homogenate method were compared to make the different dose of cell suspension injected into C57BL/6 mice. The feasibility of the improved method was determined through observing the cell count, the tumor formation ratio, the tumor formation time, tumor volume, weight and life habit. Results The method of culture LLC cells in vitro could get needed cells and its tumor formation ratio was 100 %. Trypsin digestion method and homogenate method could get less cells and its tumor formation ratio was about 80 %～90 % and 60 %～75 %. Whereas 1V collagenase method could get most cell count and its tumor formation ratio was 100 %. Conclusion IV collagenase method is a preferred method which is simple,high efficiency and make a strong base on the cancer experimental study.

Objective To investigate the differentially expressed genes of primary esophageal squamous cell carci-noma and of normal esophageal mucosa. Methods LCM-GMA-cDNA microarray was used to detect the mRNA from both the primary carcinoma and the corresponding esophageal epithelium in 15 cases of human esophageal squamous cell carcinoma (ESCC). After high-stringent washing, the cDNA microarray was scanned for the fluores-cent signals. Results Among the 886 target genes, 34 genes had significant difference in Ⅰ / Ⅱ and Ⅲ/Ⅳ group. Cell cycle regulators possibly promoting the growth of tumor cells were highly expressed in the early stages of ESCC, whereas adhesion molecules and extracellular matrix-related molecules possibly promoting invasiveness increased in the later stages. Conclusion More than one gene contributed to esophageal cancer. The profiles of gene expression will bring us chance to understanding the molecular mechanism of tumor progression and to support clinical treat-ment.

AIM:To study the effect of p21-activated protein kinase 2 (PAK2) knockdown by RNA interfer-ence on the proliferation and apoptosis of human breast cancer cells .METHODS:The short hairpin RNA ( shRNA) targe-ting PAK2 gene was designed and used for packing lentivirus in 293T cells.Human breast cancer MCF-7 cells were infected by the virus particles and PAK2 knockdown stable cell line was established by puromycin selection .The knockdown effi-ciency was assessed by Western blotting .The proliferation ability of MCF-7 cells was evaluated by CellTiter 96 AQueous and anchorage-independent growth assays .The cell apoptosis induced by staurosporine was detected by flow cytometry . RESULTS:The protein level of PAK2 was significantly suppressed after silencing of PAK2 gene in MCF-7 cells ( P<0.01).Furthermore, knockdown of PAK2 caused remarkable inhibition of the cell proliferation and colony formation (P<0.01).Staurosporine induced more apoptosis in the PAK2 knockdown cells compared with the control cells (P<0.01). CONCLUSION:Knockdown of PAK2 inhibits the proliferation of MCF-7 cells and increases the sensitivity of chemothera-peutic drug-induced cell apoptosis , suggesting that PAK2 might be a new therapeutic target in breast cancer treatment .

Objective To study the effect of gene engineering adenovirus H101 for treating esophagus carcinoma EC9706 induced by CEA gene specific silencing and to explore internal influential factor of H101 sensitivity. Methods To construct the siRNA expression vector of CEA and to inhibit the expression of CEA through RNA interference and gene transfection in EC9706 cell,stable CEA gene silencing system was set up,compared with empty vector group and non-transfected EC9706 cell,the model of athymic mouse subcutaneous transplantation tumor of human esophagus carcinoma EC9706 cell was established followed by injection with H101. The mRNA and protein expressions of CEA were detected by real time PCR and immunohistochemistry,the tumor size was measured. Results Silencing CEA gene by applying RNAi can inhibit CEA mRNA and protein expression in nude mice model with transplanted human esophageal cancer cells,there was no evident influence on tumor growth and mass oftumor. After using H101,the tumor size of interfering group was much smaller than that of empty vector group and normal control group(P

Objective:To analyze the reflux parameters of patients with gastroesophageal reflux disease (GERD) in upright position, supine position and at 2 h after meals, and to explore the cut-off value, sensitivity and specificity of the reflux parameters in different positions and at 2 h after meals in GERD diagnosis.Methods:From January 2016 to July 2020, 200 GERD patients (GERD group) and 61 non-GERD patients (control group) who visited Huazhong University of Science and Technology Union Shenzhen Hospital (Former Nanshan District People′s Hospital), were selected. All the patients of the two groups received gastroesophageal reflux disease questionnaire (GERDQ), upper gastrointestinal endoscopy, esophageal high resolution manometry and 24 h esophageal pH combined impedance monitoring. T test, non-parametric test and chi-square test were used to compare the related parameters in upright position, supine position and at 2 h after meals between two groups and within each group. Receiver oparative characteristic (ROC) curves of reflux parameters in upright position, supine position and 2 h after meals were drawn to determine the cut-off value, sensitivity and specificity in GERD diagnosis. Results:The proportion of patients with acid reflux in supine position of the control group was higher than that of the GERD group (41.0%, 25/61 vs. 8.50%, 17/200), and the difference was statistically significant ( χ2=36.53, P<0.01). In the control group, the acid reflux time in upright position, number of acid reflux, acid exposure time (AET), longest reflux time and number of weak acid reflux were more than those of in supine position in the same group (6.00 min(2.00 min, 13.50 min) vs. 0.00 min(0.00 min, 1.50 min), 16.00(8.00, 27.00) vs. 1.00(0.00, 3.00), 0.90%(0.33%, 1.88%) vs. 0.00%(0.00%, 0.30%), 2.00 min(1.00 min, 4.00 min) vs. 0.00 min(0.00 min, 1.00 min), 7.00(3.00, 11.00) vs. 1.00(0.00, 2.00), respectively) and the differences were statistically significant ( Z=5.43, 6.61, 5.06, 3.58 and 6.24, all P<0.01). In the GERD group, the acid reflux time, number of acid reflux, AET, longest reflux time and number of weak acid reflux in upright position were higher than those in supine position (51.00 min, (31.00 min, 86.75 min) vs. 8.00 min(1.00 min, 42.00 min), 60.00(48.00, 83.75) vs.6.00(2.00, 19.50), 7.30%(3.90%, 12.10%) vs. 1.50%(0.20%, 6.50%), 7.00 min(4.00, 12.00 min) vs. 4.00 min(1.00 min, 17.00 min), 1.00(0.00, 3.00) vs. 0.00(0.00, 2.00), 7.00(3.00, 12.00) vs. 0.00(0.00, 1.00), respectively) and the differences were statistically significant ( Z=7.92, 11.22, 6.90, 2.56, 5.11 and 11.76, all P<0.05). The acid reflux time, number of acid reflux, AET, longest reflux time and number of weak acid reflux at 2 h postprandial were 3.00 min(2.00 min, 9.00 min), 10.00(5.00, 18.00), 0.90%(0.40%, 1.98%), 1.00 min(0.00 min, 3.00 min), 4.00(1.50, 8.50)and 28.50 min(15.00 min, 54.75 min), 35.00(24.00, 52.00), 8.30%(4.32%, 15.83%), 6.00 min(3.00 min, 11.00 min), 4.00(2.00, 7.25), in the control and GERD groups, respectively, which were significantly higher than those in supine position in the same group ( Z=4.30, 6.33, 5.50, 3.40, 5.71 and 3.76, 9.21, 5.76, 1.97, 10.46, all P<0.05). Among 200 GERD patients, 125 patients had symptoms recorded during the 24 h esophageal pH combined impedance monitoring, the incidence of reflux symptoms in upright position was higher than that in supine position (89.6%, 112/125 vs. 65.6%, 82/125), and the difference was statistically significant ( χ2=20.71, P<0.01). The results of ROC curve analysis showed that the accuracy of acid reflux time in upright position in GERD prediction was the highest, with AUC value of 0.94 and cut-off value of 24.5 min, and the sensitivity and specificity in GERD diagnosis were 81.50% and 95.08%, respectively. The prediction accuracy of acid reflux times in upright position and AET in upright position for GERD was secondary, AUC value both were 0.93 and the cut-off value of the acid reflux number in upright position was 39.5, and the sensitivity and specificity in GERD diagnosis were 84.00% and 95.08%, respectively. The cut-off value of AET in upright position was 2.75%, the sensitivity and specificity in GERD diagnosis were 85.00% and 93.33%, respectively. The AUC value, cut-off value, sensitivity and specificity of AET at 2 h postprandial were 0.91, 4.60%, and 73.49% and 95.00%, respectively. Conclusions:Both GERD patients and non-GERD patients have more reflux in upright position, especially within 2 h after meals. The diagnostic values of acid reflux time in upright position, number of acid reflux, AET and AET 2 h after meals for GERD is high, and the AUC values are all >0.90, which can be used as a more comprehensive basis for the analysis and diagnosis of GERD.