Objective To investigate the clinical significance and biological role of G-protein coupled receptor 49(GPR49) expression in pancreatic carcinoma.Methods GPR49 expression in tumor and adjacent normal tissues of 77 patients with pancreatic cancer was compared by tissue microarray and immunohistochemistry.And then, the GPR49 expression levels in the tumor tissues of patients with different pathological grades and clinical stages were analyzed.GPR49 positive pancreatic cancer cell line CFPAC-1 was taken as cellular model.CFPAC-1 cells were stimulated with roof plate-specific spondin(RSPO)1, the ligand of GPR49, in vitro.The effect of RSPO1 on CFPAC-1 cells proliferation was evaluated with cell counting.The effect of RSPO1 on the expression of membrane molecular CD44 in CFPAC-1 cells was detected by flow cytometry.CFPAC-1 cells incubated with RSPO1 were subcutaneously implanted into nude mice.And then, the time of tumor formation and tumor size were observed.T test and single factor analysis of variance were performed for statistical analysis.Results GPR49 was widely expressed in all 77 pancreatic cancer tissues.By immunohistochemistry, the score of GPR49 expression in pancreatic cancer tissues was 9.0±2.4, which was higher than that of adjacent normal tissues (5.7±2.4), and the difference was statistically significant (t=8.995, P<0.01).There was no correlation between GPR49 expression and tumor sizes, pathological grades, lymph node metastasis, and clinical stages (all P>0.05).The results of experiments in vitro indicated that RSPO1 could promote CFPAC-1 cells proliferation and up-regulate CD44 expression in CFPAC-1 cells.Experiments in vivo demonstrated that after 30 days the tumor volume of mice implanted with RSPO1-pretreated CFPAC-1 cells was (606.0±188.0) mm3, which was larger than that of PBS-pretreated group ((364.2±83.7) mm3), and the difference was statistically significant (t=2.616, P=0.031).Conclusion GPR49 is widely expressed in pancreatic cancer cells and RSPO1/GPR49 pathway has play a role in promoting the proliferation of pancreatic cancer cells, which might be a potential target for interfering pancreatic cancer.

OBJECTIVETo observe the effect of Hydroxy Safflower Yellow A (HSYA) on the expression of osteogenic markers, such as alkaline phosphatase, Cbf(alpha)l and type I collagen, and explore the mechanism of HSYA in the prevention and treatment of glucocorticoid-induced ischemic necrosis of femoral head.

METHODSFifteen healthy and adult New Zealand white rabbits were collected and weighted 0.9 to 1.3 kg. The rabbits were injected abdominally with anesthetic drugs, then received marrow cavity puncture of tibia and anterior superior iliac spine to get bone marrow blood. Rabbits bone marrow mesenchymal stem cells (BMSCs) were separated from the bone marrow blood, cultured in vitro and passaged. The 3rd generation of BMSCs which had good growth condition were randomly divided into blank group, model group and HSYA groups with different doses. The BMSCs in model group were treated with high dose of dexamethasone to induce adipogenic differentiation of cells cultured in vitro, and inhibit osteogenic differentiation. The BMSCs in HSYA groups received high dose of dexamethasone and different concentrations of HSYA simultaneously. The blank group received not any special handling. After a week,the expressions of alkaline phosphatase, Cbf(alpha)l and type I collagen mRNA were detected.

RESULTSThe alkaline phosphatase activity was significantly decreased in BMSCs of the model group as compared with the blank group (P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also decreased significantly (P<0.01). The alkaline phosphatase activity was significantly increased in BMSCs of each HSYA group as compared with the model group (P < 0.05 or P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also increased significantly (P < 0.05 or P < 0.01).

CONCLUSIONThe mechanism of HSYA may be related to the effect of antagonism to the reduced osteogenic differentiation induced by glucocorticoid.

Alkaline Phosphatase ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chalcone ; analogs & derivatives ; chemistry ; pharmacology ; Collagen Type I ; genetics ; metabolism ; Core Binding Factor alpha Subunits ; genetics ; metabolism ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Glucocorticoids ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Osteogenesis ; drug effects ; Rabbits

Objective To study the effects of hSSTR2 gene in vitro transfection on differential proteins expression in pancreatic cancer cell line Panc-1 and search new sensitive therapeutic targets of pancreatic cancer. Methods The full length hSSTR2 cDNA was introduced into pancreatic cancer cell line Panc-1 by adenovirus vector ( Ad. CMV. hSSTR2. GFP) mediated transfection. The differential expressed proteins between the hSSTR2 transfection group, vector control and mock control were isolated and screened by 2D-DIGE analysis. Protein identification was performed by peptide mass finger printing with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF). Results The hSSTR2 gene was transfected into Panc-1 pancreatic cancer cells in vitro successfully, and fluorescence difference protein expression patterns were established between hSSTR2 negative and positive expression of Panc-1 cell. Analysis by DeCyder v6.5 software showed a total of 18 protein spots ( > 1.3-fold) and these protein spots were identified by mass spectrometry as 13 proteins. Proteins with lower abundance levels included GMP synthase, stress induced phosphoprotein 1, glutamate dehydrogenase 1, Septin-11, vimentin, Isocitrate dehydrogenase [NAD] subunit alpha, Import inner membrane translocase subunit TIM50. Proteins with high abundance levels included Elongation factor 1-alpha-1, Isoform M2 of Pyruvate kinase isozymes M1/M2, Enoyl-CoA hydratase,tripartite motif-containing 28 protein, Mitofilin, HSP105. Conclusions The proteins expression changed after hSSTR2 gene in vitro transfection in Panc-1 cells, and the function of difference proteins involved the process of metabolism of sugar, fat and nucleic acid, and the regulation of cell growth. The present study paved the way for searching new sensitive therapeutic targets of pancreatic cancer.

Mounting evidence has shown that side population (SP) cells are enriched for cancer stem cells (CSCs) responsible for cancer malignancy. In this study, SP technology was used to isolate a small subpopulation of SP cells in human gallbladder cancer cell line GBC-SD, and SP cells which had superior potential for proliferation in vitro and tumorigenesis in vivo were identified. Importantly, the abundance of GBC-SD SP cells was increased by a transforming growth factor-β (TGF-β)-induced epithelial-mesenchymal transition (EMT), and this effect was accompanied with a strong up-regulation of ABCG2 mRNA expression, and a decreased sensitivity to mitoxantrone. SP cells were restored upon the removal of TGF-β and the reversion of the cells to an epithelial phenotype, and smad3-specific siRNA reduced SP abundance in response to TGF-β. In conclusion, TGF-β-induced EMT by smad-dependent signaling pathway promotes cancer development and anti-cancer drug resistant phenotype by augmenting the abundance of GBC-SD SP cells, and a better understanding of mechanisms involved in TGF-β-induced EMT may provide a novel strategy for preventing cancer progression.

Postoperative pancreatic leakage remains a persistent problem after pancreaticoduodenectomy.For patients with a soft and nonfibrotic pancreas,double binding continuous hemstitch suture is an optimal method for anastomosis.From January 2011 to June 2012,92 cases of periampullary carcinoma with a soft pancreas underwent pancreaticoduodenectomy,and then a modified technique of pancreaticogastrostomy was performed with 2 continuous hemstitch sutures placed in the mucosal and seromuscular layers of the posterior gastric wall,respectively.The median time for pancreaticogastrostomy was 12 minutes,and only 15 patients had postoperative complications.Two patients developed pancreatic leakage(1 grade A and 1 grade B)postoperatively.Pancreaticogastrostomy with double binding continuous hemstitch sutures is a simple and safe reconstruction procedure for patients with a soft and fragile pancreas who received pancreaticoduodenectomy.

Objective To investigate the expression of Bcl-2,survivin and pancreatic cancer stem cells markers Oct-4 and ABCG2 in pancreatic cancer cells resistance to chemoradiotherapy.and explore its mechanism.Methods Concurrent ehemoradiotherapy was used to obtain pancreatic cancer cells resistant to chemoradiotherapy,the pancreatic cancer cells without chemoradiotherapy treatment were used as control.Western-blot was applied to detect the expression of Bcl-2,survivin,Oct-4,ABCG2.Results The expression of Bcl-2 was 0.7955±0.0326,0.5718±0.0212,0.6137±0.0382 and 0.8733±0.0461,respectively;the expression of survivin protein was 0.8207±0.0490,0.6973±0.0211,0.7967±0.0346 and 0.8013±0.0398,respectively;the expression of Oct-4 protein was 0.8728±0.0177,0.7861±0.0139,0.4794±0.0932 and 0.4216±0.1043,respectively;the expression of ABCG2 protein was 0.7810±0.1370,0.4957±0.1126,0.6102±0.1358 and 0.4670±0.1274,respectively.in resistant pancreatic cancer cells of SW1990,BxPC3,pc3,jf305 cell line.The corresponding values in the control group were 0.4723±0.018,0.2954±0.0103.0.3587±0.0201 and 0.2718±0.0136;0.4717±0.0274,0.3587±0.0113,0.3891±0.0147 and 0.3326±0.0124;0.6053±0.0142,0.4236±0.0086.0.2385±0.0671 and 0.1985±0.0582;0.3156±0.0582.0.2360±0.0423,0.2813±0.0512 and 0.1808±0.a0370.The expression of all the four proteins significantly increased after ehemoradiotherapy(P＜0.05).Conclusions Pancreatic cancer cells resistant to chemoradiotherapy may contain cancer stem cells.

Objective To detect the role of differentially expressed microRNA (miRNA) in human cholangiocarcinoma and explore their effects on apoptosis and invasiveness of cholangiocarcinoma.Methods The differential expression of miRNA in 3 cholangiocarcinoma patients was detected by miRNA array.The expressions of miR-200a,miR-200b,miR-200c,and miR-141 in human cholangiocarcinoma tissues and normal bile duct tissues were detected by real-time PCR.After transfection with miR-200b mimic,apoptosis and invasiveness of human cholangiocarcinoma cell line QBC939 was evaluated by Annexin-V-FITC dyeing and Transwell assay.Results Comparad with normal bile duct tissues,the number of differential miRNAs in cholangiocarcinoma was 21,including 15 up regulated and 6 down regulated.The expressions of miR200a,miR-200b,miR-200c,and miR-141 in human cholangiocarcinoma tissues were significantly lower than levels in normal bile duct tissues.The invasive ability of QBC939 was decreased after miR-200b mimic transfection.The apoptosis cell number of QBC939 was increased after miR-200b mimic transfection.Conclusion These results indicate that the expression of miRNA is different between cholangiocarcinoma and normal bile duct tissues.Moreover,miR-200a,miR-200b,miR-200c,and miR-141 are likely involved in the invasion and metastasis of cholangiocarcinoma and have potential as a diagnostic and prognostic marker.

Objective To evaluate a modified technique for digestive tract reconstruction after pancreaticoduodenectomy(PD).Methods 171 admitted patients were enrolled from January 2012 to January 2014 at our department.According to the preoperative CT scan and intraoperative exploration,pancreaticogastrostomy was performed in cases of soft pancreas texture,while pancreaticojejunostomy was performed in fibrotic pancreas after PD.Bypassed biliary-pancreatic reconstruction were applied on all cases.Results For the digestive tract reconstruction after PD,92 patients underwent pancreaticogastrostomy,79 patients underwent pancreaticojejunostomy.The median time for the surgery was 240.0 minutes (ranging from 186 to 414 min).Operative mortality was zero,and morbidity was 18.1％ (n =31),including hemorrhage (n =4),biliary fistula (n =3),pulmonary infection (n =2),adipose liquefaction and operative incision infection (n =0),delayed gastric emptying (DGE) (n =6),abdominal abscess (n =4).Fout patients developed a pancreatic fistula (type A in 2,type B in 2).Conclusions Modified pancreaticogastrostomy,pancreaticojejunostomy and biliary-pancreatic bypass is safe for digestive tract reconstruction after pancreaticoduodenectomy.

Objective: To investigate the delay in identification of pulmonary tuberculosis and influencing factors among children and adolescents in Jiaxing City, Zhejiang Province from 2013 to 2022, so as to provide the reference for targeted prevention and control measures. Methods: The information of pulmonary tuberculosis patients in Jiaxing City from 2013 to 2022 were captured from the Tuberculosis Information Management System of Chinese Disease Control and Prevention Information System, including demographics, diagnosis, treatment and etiological results. The delay in identification of pulmonary tuberculosis was analyzed among children and adolescents, and the factors affecting the delay in identification of pulmonary tuberculosis were identified using a multivariable logistic regression model. Results: A total of 2 407 pulmonary tuberculosis cases were reported among children and adolescents in Jiaxing City from 2013 to 2022, including 1 522 males (63.23%). The median age was 21.00 (interquartile range, 4.00) years. There were 410 students (17.03%), and 1 856 cases with non-local household registration (77.11%). There were 596 cases with delay in identification of tuberculosis (24.76%), 895 cases with delay in healthcare-seeking (37.18%) and 128 cases with delay in definitive diagnosis (5.32%). Multivariable logistic regression analysis that children and adolescents who occurred symptoms in the first quarter (OR=1.684, 95%CI: 1.261-2.249), were diagnosed first in county-level medical institutions (OR=3.800, 95%CI: 2.898-4.983) and had positive results of etiological testing (OR=1.534, 95%CI: 1.255-1.874) were more likely to delay in identification of pulmonary tuberculosis. Conclusions The delay in identification of pulmonary tuberculosis is associated with the time of symptom onset, the level of medical institution making first diagnosis, and the results of etiological testing. It is suggested to reinforce the publicity of pulmonary tuberculosis prevention and control, expand the coverage of screening programs and improve the diagnosis capability of medical institutions.