Objective To explore the influence and clinical significance of chemotherapy on the expressions of micro-vessel density (MVD) and vascular endothelial growth factor (VEGF) in pediatric difficult removal solid tumor.Methods From Jan.2002 to Jan.2012,totally 80 cases of the hospitalized pediatric patients with solid tumor (Ⅲ b-Ⅳb stage) were enrolled in this study in Department of Pediatric Surgery of the Second Hospital of Hebei Medical University.All of the tumors were identified as difficult removal malignant tumor by imageology and physical examinations.After biopsy of the tumors,these patients were divided into neuroblastoma group (25 cases),hepatoblastoma group (25cases) and Wilms' tumor group (30 cases).All patients in these 3 groups underwent chemotherapy respectively according to their different pathologic findings,thereafter,the tumor resections were performed.The resection rates of tumor in each groups were recorded and the expressions of MVD and VEGF between preoperative biopsy and postoperative specimens were detected and compared by immunohistochemistry in different groups.Results With chemotherapy,the resection rate of neuroblastoma was 80.00％ (20/25 cases),hepatoblastoma was 92.00％ (23/25 cases) and Wilms' tumor was 96.67％ (29/30 cases).There was no significant difference among them (x2 =4.286,P =0.117).Before chemotherapy,the positive expressions of VEGF in neuroblastoma,hepatoblastoma and Wilms' tumor were 80.00％ (20/25 cases),90.00％ (18/20 cases) and 83.33％ (25/30 cases) respectively,at the same time,the values of MVD were 48.39 ± 15.87,52.16 ± 16.12 and 46.82 ± 19.07.After chemotherapy,the expressions of VEGF and MVD in neuroblastoma,hepatoblastoma and Wilms' tumor were 52.00％ (13/25 cases),45.00％ (9/20 cases),30.00％ (9/30 cases) and 31.31 ± 14.21,25.19 ± 15.31 and 29.06 ± 13.05.Compared with the expressions of them in each group before chemotherapy,significant differences were found (all P ＜ 0.05).Conclusions Chemotherapy can efficiently promote the resection rate for pediatric difficult removal solid tumor.MVD and VEGF can be considered as the markers in evaluating the outcomes of chemotherapy and promoting the resection rate of tumors.

Objective:To investigate the differential expression of cyclooxygenase (COX) gene between familial aggregated atherosclerotic large-artery atherosclerosis(LAA) cerebral infarction patients and sporadic LAA cerebral infarction patients.Methods:Twenty-five patients with familial aggregation LAA cerebral infarction (familial aggregation LAA group) and 25 sporadic LAA cerebral infarction patients (sporadic LAA group) were collected.Another 25 patients with non-cardiovascular and cerebrovascular diseases were selected as control group.Real-time quantitative PCR and ELISA were used to detect COX-1 and COX-2 protein in the supernatant of samples.Results:The expression level of COX-1 gene was 0.5436±0.2737, 0.2400±0.1656 and 0.8340±0.3799 in the familial aggregation, sporadic LAA cerebral infarction and control groups.Compared with control group, COX-1 gene expression in sporadic LAA cerebral infarction group and familial aggregation LAA cerebral infarction group was significantly down-regulated, the differences were significant( P<0.05). The expression level of COX-2 gene was 1.3672±0.8249, 1.3932±0.7158 and 0.7212±0.9623 in the familial aggregation, sporadic LAA cerebral infarction and control groups.Compared with the normal control group, the expression of COX-2 gene in familial aggregation LAA cerebral infarction group and sporadic LAA cerebral infarction group increased significantly( P<0.05). The expression of COX-1 protein was 2.9956±0.5672, 2.5572±1.0289 and 2.6721±0.7944 in the familial aggregation, sporadic cerebral infarction and control groups.The expression of COX-2 protein was 16.63±4.06, 16.86±7.93 and 15.94±5.94 in the familial aggregation, sporadic cerebral infarction and control groups.There was no significant difference in the relative expression of COX-1 and COX-2 proteins among familial aggregation LAA cerebral infarction group, sporadic LAA cerebral infarction group and control group(all P>0.05). Conclusion:There is significant difference in COX-1 gene expression between patients with familial LAA and sporadic LAA cerebral infarction.

Objective:To investigate the effect and quality of life after anterior chest wall reconstruction with titanium plate and mesh due to manubrium tumor resection.Methods:A retrospective analysis was performed from January 2012 to December 2016, a total of 13 patients with primary(11cases) or secondary(2 cases) manubrium tumor, aged 41-72 years(medium aged as 56), underwent oncological resection, following by immediate chest wall reconstruction with titanium plate and titanium mesh. The quality of life was evaluated by EuroQol five dimensions questionnaire (EQ-5D) and EQ-5D visual analogue scale (ED-VAS) before and 3 months after operation. Computed tomographic scan was taken 3 months after operation.Results:All the patients received successful operation. During the perioperative period, 12 cases recovered well without any complications, 1 patient experienced mild paradoxical movement within 9 days after operation and then recovered. The median follow-up period after surgery was 25 months (range from 7 to 41). Computed tomographic scans for the 13 patients showed neither dislocation nor deformation of the titanium plate and mesh 3 months after operation. One patient with chondrosarcoma died as a result of relapse 11 months after surgery. In pain/discomfort dimension, the pre-operation score was 1.85±0.80 and the post-operation score was 1.15±0.38, showing a significant difference( P=0.013). There was no significant difference in scores of other four dimensions between pre- and post-operation( P>0.05). The total score of EQ-5D decreased significantly after the operation (7.08±2.02 vs 5.45±0.52, P=0.010). The pre- and post-operation ED-VAS score was 85.69±7.58 and 92.54±2.53, with a significant difference( P=0.008). Conclusions:The immediate reconstruction of chest wall with titanium plate and mesh after oncological manubrium resection is effective and safe with improvement of the quality of life.

Objective To establish breast cancer organoids for a long time and to characterize their molecular ex-pression and test their sensitivity to chemotherapy drugs.Methods Breast cancer specimens were digested with col-lagenase to release cancer cells for organoid culture.The organoids were characterized by morphology and ER,PR,HER-2,CK expression by technology of histoimmunofluorescence.Among them,three were tested for paclitaxel,doxorubicin and cisplatin sensitivity.Results A total of 44 cases of breast cancer organoids were established,all of which showed dense spherical-like growth and ER,PR,HER-2,CK,matching their clinical counterpart.Breast cancer organoids were sensitive to chemotherapy drugs paclitaxel,doxorubicin and cisplatin.Conclusions Organoid model,as a new in vitro may reproduce the pathophysiology features with heterogeneity.

Objective:To investigate the effect and quality of life after anterior chest wall reconstruction with titanium plate and mesh due to manubrium tumor resection.Methods:A retrospective analysis was performed from January 2012 to December 2016, a total of 13 patients with primary(11cases) or secondary(2 cases) manubrium tumor, aged 41-72 years(medium aged as 56), underwent oncological resection, following by immediate chest wall reconstruction with titanium plate and titanium mesh. The quality of life was evaluated by EuroQol five dimensions questionnaire (EQ-5D) and EQ-5D visual analogue scale (ED-VAS) before and 3 months after operation. Computed tomographic scan was taken 3 months after operation.Results:All the patients received successful operation. During the perioperative period, 12 cases recovered well without any complications, 1 patient experienced mild paradoxical movement within 9 days after operation and then recovered. The median follow-up period after surgery was 25 months (range from 7 to 41). Computed tomographic scans for the 13 patients showed neither dislocation nor deformation of the titanium plate and mesh 3 months after operation. One patient with chondrosarcoma died as a result of relapse 11 months after surgery. In pain/discomfort dimension, the pre-operation score was 1.85±0.80 and the post-operation score was 1.15±0.38, showing a significant difference( P=0.013). There was no significant difference in scores of other four dimensions between pre- and post-operation( P>0.05). The total score of EQ-5D decreased significantly after the operation (7.08±2.02 vs 5.45±0.52, P=0.010). The pre- and post-operation ED-VAS score was 85.69±7.58 and 92.54±2.53, with a significant difference( P=0.008). Conclusions:The immediate reconstruction of chest wall with titanium plate and mesh after oncological manubrium resection is effective and safe with improvement of the quality of life.

Irritable bowel syndrome （IBS） is a functional gastrointestinal disease， but it often causes extreme gastrointestinal discomfort and prolonged illness， which seriously affects the quality of life of patients. The global incidence rate is increasing year by year. Clinically， western medicine mainly uses oral antispasmodics， secretagogues， and antidepressants， but there are many disadvantages such as adverse reactions and poor long-term efficacy. Therefore， finding an efficient and safe treatment method is an urgent problem to be solved. A large number of studies have shown that traditional Chinese medicine has definite curative and long-lasting effects on the treatment of IBS， which has become a hot research direction in recent years. By searching Chinese and foreign literature， it is found that electroacupuncture， moxibustion， Chinese medicine monomers， and compound decoctions are the main methods in the mechanism research of traditional Chinese medicine in the treatment of IBS-related pathways， and their signaling pathways involve nuclear transcription factor kappa B （NF-κB）， transient receptor potential vanillin subfamily 1 （TRPV1）， 5-hydroxytryptamine （5-HT）， mitogen-activated protein kinase （MAPK）， and so on. Traditional Chinese medicine can repair intestinal inflammation， reduce visceral sensitivity， enhance intestinal mucosal barrier， and regulate intestinal motility by regulating this series of signaling pathways， thereby playing an important role in the treatment of IBS with multi-level， multi-link and multi-target characteristics. Based on the cell signaling pathways， this paper reviewed the research progress on the mechanism of traditional Chinese medicine in the treatment of IBS， hoping to provide theoretical support and diagnosis and treatment ideas for the clinical treatment of IBS with traditional Chinese medicine.

Objective To establish human and mouse pancreatic cancer cell strains stably expressing Cas9 protein,green fluorescent protein,red fluorescent proteins,luciferase-tdTomato,and to validate the activity of luciferase and gene editing of Cas9 function for pancreatic cancer research using luciferase and CRISPR/Cas9 system.Methods In human pancreatic cancer cells(AsPC-1,CFPAC-1,HPAC,BxPC-3,HS 766T,MIA PaCa-2,PANC-1,and SW 1990),and mouse pancreatic cancer cell(Pan02),the cells were infected with Cas9-expressing plas-mid pLv-EF1α-Cas9m1.1-Puro,and single-cell clones were selected for culture and expansion.After extracting the total protein,Western blot verified the expression level of Cas9;Infected with fluorescent protein expression plasmids pLv-EF1α-EGFP,pLv-EF1α-mCherry,pLv-EF1α-tdTomato,pLv-EF1α-Luc2-tdT,and selected single cell clones stably expressing fluorescent proteins were cultured and amplified under fluorescence microscope.Cas9 stable expression cell line was selected to be infected with pLv-EF1α-Luc2-tdT,and the mono-clonal culture of stable expression of fluorescent proteins was selected for expansion under fluorescence micro-scope.One of the cell lines were selected to be infected with Lv-EF1a-mCherry,and the mCherry-positive cells were sorted out by flow cytometry,and then the guide RNA of mCherry gene was then infected by lentivirus to tar-get the mCherry gene,and after cell expansion,mCherry knockdown was detected by fluorescence microscope observation and flow cytometry;5 BALB/c Nude mice were subcutaneously inoculated with MIA PaCa-2-Luc2-tdT cells(1.0×107/cells each),and imaged in vivo after 36 days.Results 48 human pancreatic cancer cell strains with stable Cas9 expression were screened(including 23 cells expressing Cas9m1.1,25 cells expressing Cas9m1.1-Luc2-tdT),33 pancreatic cancer cell strains with stable expression of fluorescent proteins were screened(8 cells expressing EGFP,7 expressing mCherry,and 9 each expressing Luc2-tdT and tdTomato).Cells expressing mCherry and Cas9 were infected with mCherry gRNA and mCherry was knocked down.In vivo imaging showed that both bioluminescence and fluorescence luminescence were present in MIA PaCa-2 cells ex-pressing Luc2-tdT.Conclusions 33 pancreatic cancer cell strains with stable expression of fluorescent proteins are successfully established,in which the Luc2-tdT-expressing cell strains have luciferase activity;48 pancreatic cancer cell strains with stable expression of Cas9 are successfully established,and the Cas9 protein has gene edi-ting activity,gene editing activity varies depending on the original cell strains.

Objective: To explore the characteristics of cognitive processing over time in recognition of different emotional faces in patients with depression. Methods: Totally 29 patients with depression(patient group) and 30 normal control (control group)were selected.Subjects freely viewed positive, sad, threatening and neutral facial stimuli.Eye movement data were collected during the process of watching, and the differences of attention bias in time when the two groups of subjects recognized the four emotional faces were compared.The temporal variation trend of attention between the two groups was analyzed. Results: There was no significant difference in the fixation duration between the patient group and the control group in 0-2 s((10.39±1.83)ms, (9.63±1.28)ms), 2-4 s((12.61±2.34)ms, (11.21±3.00)ms), 4-6 s ((12.26±3.11)ms, (10.47±4.06)ms)(all P>0.05). Within 6-8 s((12.64±3.42)ms, (8.90±4.02)ms), 8-10 s((12.55±4.19)ms, (9.36±4.02)ms), the fixation duration in the patient group was significantly longer than that in the control group (P<0.01). The fixation time of the positive faces in the patient group in 0-2 s ((12.07±2.65)ms, (14.50±3.54)ms), 2-4 s((10.69±3.26)ms, (14.66±5.25)ms), 4-6 s((10.11±4.24)ms, (15.43±8.02)ms), 6-8 s((9.39±5.12)ms, (16.88±9.60)ms), 8-10 s ((9.45±4.94)ms, (16.63±10.15)ms) was significantly lower than that in the control group (all P<0.01). Conclusion Patients with depression tend to pay attention to negative stimuli in the later stage of cognitive processing, while the attention to positive stimuli decreases in the early stage of cognitive processing.Cognitive processing of emotional stimuli in patients with depression is manifested as increased attention duration of negative stimuli and decreased attention duration of positive stimuli.

Objective: To evaluate the diagnostic efficiency and accuracy of endoscopic ultrasonography (EUS) on vascular involvement of pancreatic cancer. Methods: Patients, suspected pancreatic cancer with vascular involvement by CT scan in Huashan Hospital, Fudan University from January 2014 to March 2019, were enrolled prospectively in the study. EUS was performed to evaluate the vascular involvement compared with surgical pathological results. Results: A total of 132 patients with pancreatic cancer were enrolled in the study, and they all underwent EUS observation and radical resection with vessels resection. There were 103 cases of cancer in pancreatic head, 19 cancers in pancreatic neck and 10 cancers in distal pancreas. The diagnostic sensitivity, specificity and accuracy of EUS was 97.4% (113/116), 81.2% (13/16), and 95.5% (126/132), respectively for pancreatic cancers with vein involvement; while was 33.3% (2/6), 90.0% (81/90), and 86.5% (83/96), respectively, for pancreatic cancers with superior mesentery artery involvement. Conclusion EUS may play a key role in diagnosis of vascular involvement of pancreatic cancer, and be helpful for the surgical decision marking.