ObjectiveTo explore the tumor igenic property of side population cells (SP) from human gallbladder carcinoma cell line GBC-SD. Methods SP and non-SP cells were isolated from GBC-SD staining with Hoechst33342 dye by fluorescence-activated cell sorting (FACS). The soft agar clonal assay and xenograft assay were performed to characterize tumorigenic property of side population cells in vitro and in vivo, respectively. The percentage of SP cells was analyzed by FACS in 5 hu man gallbladder carcinoma specimens. ResultsThe percentage of SP cells accounted for approximately 0.87 ％ of GBC-SD cells. The clone-formed rates of SP was more frequent than that of non-SP cells (14.74％ ± 3.53％ vs 5.17％ ± 1.05％), there was statistically significant difference (t =2.75,P＜0. 05). SP cells could generate tumors with as few as 5 × 103 cells (four of seven animals), whereas at least 1 × 105 non-SP cells were needed to form a tumor (one of seven animals). Re-analysis of SPderived tumors by FACS showed that SP cells under in vivo conditions also have the capacity to regenerate the SP and non-SP fractions. Besides, analysis of Hoechst33342 revealed s small fraction of SP cells, ranging from 0. 27％ to 2.3％ in gallbladder carcinoma specimens. ConclusionSP cells from GBC-SD are highly tumorigenic similar as the cancer stem cells.

Objective To investigate the drug resistance of side population cells in human gallbladder cancer cell line GBC-SD and explore its mechanism. Methods Drug sensitivity assays of 5chemotherapeutic agents were performed on side population cells (SP) and non-SP cells of GBC-SD.GBC-SD was cultured and then treated with the chemotherapeutic agent gemcitabine. The frequency of SP by FACS was measured. RT-PCR and Western blotting were used to detect the expression of AB-CG2 in both the SP and the corresponding non-SP subsets. Results After 1 d treatment with 4 chemotherapeutic agents (gemcitabine, cisplatin, 5-fluorouracil and mitoxantrone) in IC50 concentration to GBC-SD cell line, the reproductive ability of SP was higher than that of non-SP (P<0.05). However, statistical significance was not achieved when compared with epirubicin (P>0.05). The percentage of SP in GBC-SD treated with chemotherapeutic agent gemcitabine after 3 weeks was sharply elevated by FACS (8.02% ±0.13% vs 0.62% ±0.08%, P<0.05), and the expression of ABCG2mRNA and protein were increased in SP as compared with non-SP. Conclusion SP from human gallbladder cancer cell line GBC-SD, like stem cell, showed a heighten resistance to drugs. Increased expression of ABCG2 was largely responsible for the multi-drug resistance.

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 clinical efficacy of open reduction and internal fixation with dynamic anterior plate-screw system for quadrilateral area ( DAPSQ ) in the treatment of elderly patients with acetabular fracture involving medial displacement of the quadrilateral surface. Methods Between January 2007 and December 2014, a series of 18 senile patients with acetabular fractures involving medial displace-ment of the quadrilateral surface were treated at our department. They were 13 men 5 women, with a mean age 67. 8 years ( range, from 61 to 78 years ) . By the Judet-Letournel classification, there were 4 anterior column fractures, 5 anterior column plus posterior hemitransverse fractures, 8 double column fractures and one T-shaped fracture. The delay from injury to surgery averaged 6. 5 days. All were treated with the technique of DAPSQ. Screw internal fixation of the quadrilateral area followed plate moulding via a single ilioinguinal ap-proach. Fixation with lag screws for the posterior column was added if necessary. Results The 18 pa-tients were followed up for 12 to 35 months ( mean, 26 months ) . By the Matta criteria for fracture reduction, 9 cases were rated as excellent, 6 as good and 3 as poor. Bone healing was achieved within 2 to 4 months( mean, 3. 5 months ) . By the modified Merle d'Aubigné & Postel criteria, the affected hips scored 9 to 18 points ( mean, 16. 3 points ) at the final follow-ups, giving 7 excellent, 7 good, 2 fair and 2 poor cases. By the Harris evaluation, the affected hips scored 58 to 98 points ( mean, 87. 6 points ) , giving 7 excellent, 8 good, one fair and 2 poor cases. The postoperative complications included urinary tract infection in one, lesion of lateral femoral cutaneous nerve in one, heterotopic ossification ( Brooker Grade Ⅰ) in one, and traumatic osteoarthritis in 5 cases 2 of whom had to receive total hip arthroplasty. Conclusion The technique of DAPSQ is suitable for the treatment of acetabular fractures involving medial displacement of the quadrilateral surface in elderly patients, because it can provide and maintain stable fixation of the fracture in the quadri-lateral area and preserve hip joint function, leading to fine outcomes.

Objective To optimize the extraction technology of total falvonoids from Hypericum sampsonii .Methods On the basis of single factor test ,solid‐liquid ratio ,ethanol concentration ,extraction time ,extraction temperature and extrac‐tion times on the extraction rate were studied by taking the content of total falvonoids as indexes .Orthogonal test was conduc‐ted to establish the extraction technology of total falvonoids from Hypericum sampsonii .Results The influence degree of fac‐tors on the extraction rate of total falvonoids was in the order of ethanol concentration ,solid‐liquid ratio ,extraction times ,and temperature .The optimum extraction condition was confirmed as follows:ethanol concentration of 70% ,extraction tempera‐ture of 60 ℃ ,and extraction times of 3 × 1 .5 h ,solid to liquid ratio of 1:20 .Under this condition ,the average extraction rate of total falvonoids reached 2 .38% .Conclusion The test was reasonable in design and credible in findings ,and was easily oper‐ated ,which could provide scientific basis for the batch extraction of total flavonoids from Hypericum sampsonii .

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.

Objectives To isolate cancer stem cells (CSCs) in pancreatic cancer cell lines PANC1 and ASPC-1 with serum-free medium( SFM ), and to detect the expression of miR-590-3p in CSCs. Methods PANC1 and ASPC-1 cells was cultured in serum-free medium. The monoclonal formation, differentiation and cell cycle, half inhibitory concentration ( IC50 ), and the expression of the surface markers CD24 + , CD44 + were detected. qRT-PCR was used to detect the expression of miR-590-3p. Results After SFM culture, (0.94 ±0.53 ) ％ of ASPC-1 and (0.57 + 0. 12 ) ％ PANC1 survived, and they formed spheres, and could continuously passage in vitro. Cell spheres differentiation recurred when serum was supplemented in SFM. The G0/G1 stage proportion, CD24+ , CD44 + , CD24+ CD44+ cells proportion, IC50 in ASPC-1 cell were (75.3 ± 5.4)％,0.96％ ～ 2.01％, 27.52％ ～ 34.47％, 0.35％ ～ 0.44％ and (224.37 ± 5.71 ) μg/ml, which were significantly higher than that those in parent cell [ (43.7 ± 3.8 ) ％, 0. 38％ ～ 0.42％, 17.65％ ～ 18.25％,0.05％ ～0.08％, (11.43 ±2.10)μg/ml, P＜0.05]. The G0/G1 stage proportion, CD24+ ,CD44+ ,CD24 +CD44 + cells proportion, IC50 in PANC 1 cell were ( 80. 1 ± 4.7) ％, 5.31％ ～ 9.84％, 72.05％ ～ 93.06％,4.91％ ～5.21％, (296.58±4.27) μg/ml, which were significantly higher than that those in parent cell [ (46.1 ±5.3)％, 4.09％ ～4.97％, 47.71％ ～55.66％, 1.48％ ～2.63％, (26.17 ±3.81)μg/ml, P＜0.05]. The expression of miR-590-3p in ASPC-1, PANC1 spheres was 4.67 and 4.52 times higher than the expression in parent cell lines. Conclusions Pancreatic cancer cell spheres can be isolated from ASPC-1, PANC1 by culture with SFM. miR-590-3p is up-regulated and may play an important role in regulating biological characteristics of pancreatic cancer stem cells.

Objective To study the influence of eleutheroside E (ELE)on the learning and memory abilities of mice in-duced by para-chlorophenylalanine (PCPA) and on hippocampus′monoamine neurotransmitters of serotonin(5-HT) ,5-hydrox-indole acetic acid (5-HIAA) ,dopamine (DA) and norepinephrine (NA) .Methods Balb/c mice were randomized into control group ,model group ,positive group and three treatment groups (high ,moderate and low dose group) .The mouse injury model was established by PCPA abdominal injection .The mice′s abilities of learning and memory were detected in the Morris water maze test after 14 days of prevention medicine .The contents of 5-HT ,5-HIAA ,DA and NA in mice′s hippocampus were de-tected by Elisa kit .Results The behavior results showed that ,compared with control group ,the times across the platform by model group decreased significantly (P<0 .01) .The times across the platform by high and moderate dose groups increased sig-nificantly (P<0 .01) after the administration of ELE .The contents of 5-HT and 5-HIAA of the model group were lower than those of the control group (P<0 .01) .The contents of NA was lower too (P<0 .05) .The contents of 5-HT and 5-HIAA of different dose groups were much higher than those of the model group .Conclusion The enhancement of learning and memory ability in the mouse injury model established by PCPA might be related with the improvement of the contents of 5-HT ,5-HIAA ,DA ,NA in mice′s hippocampus by ELE .

OBJECTIVETo explore the role of ZNF217 in regulating cell proliferation, migration and invasion in glioma cells.

METHDOSA lentivirus-mediated shRNA-ZNF217 vector was infected into glioma U251 cells, and the interference efficiency was examined by Western blotting. MTT assay, flow cytometry, Transwell assay, and Boyden chamber assay were used to analyze the changes in cell proliferation, migration and invasion. Western blotting was used to detect the changes in ZNF217-related genes in the cells.

RESULTSshRNA-ZNF217 transfection significantly inhibited the expression of ZNF217 in U251 cells and suppressed the cell migration, invasion, growth, and cell cycle transition. ZNF217 knockdown downregulated the expression of pPI3, pAKT, C-Myc, and the mesenchyme biomarker N-cadherin, and stimulated the expression of the epithelium biomarker E-cadherin.

CONCLUSIONZNF217 promotes cell migration, invasion, and growth by activating PI3K/AKT signal to upregulate C-Myc and by modulating the genes associated with epithelial-mesenchymal transition in glioma cells.

Cadherins ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Epithelial-Mesenchymal Transition ; Genetic Vectors ; Glioma ; pathology ; Humans ; Lentivirus ; Neoplasm Invasiveness ; RNA, Messenger ; RNA, Small Interfering ; genetics ; Trans-Activators ; genetics ; Transfection