Objective To investigate the effect of ketamine on the mitochondrial function of rat neurons subjected to anoxia. Methods Primarily cultured rat hippocampal neurons were seeded in culture dishes (35 mm in diameter) at the density of 5×105-1×106 cells∕ml, and divided into 3 groups (n=11 each) using a random number table: control group, anoxia group and ketamine group. The neurons were exposed to 90% N2 plus 10% CO2 50 ml∕min for 5 min in anoxia group. In ketamine group, ketamine was added to the culture medium with the final concentration of 20 μmol∕L at 1 h before anoxia, and then the neurons were exposed to 90% N2 plus 10% CO2 50 ml∕min for 5 min. After the end of treatment in each group, the dead neurons were detected using trypan blue staining, the ATP content was determined by ATP bioluminescence assay, and mitochondrial membrane potential was measured by rhodamine 123 staining. Results Compared with control group, the mortality rate of hippocampal neurons was significantly in?creased, and the ATP content and mitochondrial membrane potential were significantly decreased in anoxia group and ketamine group ( P<0.05) . Compared with anoxia group, the mortality rate of hippocampal neu?rons was significantly decreased, and the ATP content and mitochondrial membrane potential were signifi?cantly increased in ketamine group (P<0.05). Conclusion The mechanism by which ketamine amelio?rates anoxia?induced damage to rat neurons is related to improved mitochondrial function.

BACKGROUNDCalcaneal fractures are relatively common but complex, while the treatment of these fractures is still controversial. The objective of this study was to compare the biomechanical stability of minimally invasive, locking, and non-locking plating systems used for fixing Sanders type II and type III calcaneal fractures. Finite element analysis was used for comparing the three types of plating systems. This study helped in guiding the clinical management of calcaneal fractures.

METHODSThe CT data acquired from a volunteer was used to construct a finite element model of calcaneus. The fracture lines were made according to Sanders type II (A, B, C) and type III (AB, AC, BC) and six calcaneal fractures models were constructed accordingly. Three fixation methods used in clinical treatment were simulated by design tools (minimally invasive plating with percutaneous screws, Y-size non-locking plating, and locking plating) and then fixed to these fracture models with the help of clinical principles. The single minimally invasive plating fixation was also tested to compare the function of percutaneous screws. Then, the neutral force was applied on established methods. The displacement and presses distribution condition in various plating models were analyzed and compared.

RESULTSThe levels of displacement and compressive press on the bone around fixation or strains on plating were within the physiological limit, indicating that the three methods were suitable for application. Compared with the locked and unlocked plates, the minimally invasive plating caused a relatively lower displacement without the high stress in bone or fixation. The minimum stress in fixation was observed in locking plate as the distribution of stress was concentrated in the anterior part of locking plate.

CONCLUSIONSThe minimally invasive plating method could be a better alternative method in treating Sanders type II and type III calcaneal fractures. The observed stiffness of locking plate was not obviously stronger than unlocked plate.

Adult ; Ankle Injuries ; surgery ; Bone Plates ; Calcaneus ; injuries ; surgery ; Finite Element Analysis ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Young Adult

Objective To screen and analyze Dengue virus-derived vsRNAs and piRNAs in Aedes albopictus.Methods Female adults Aedes albopictus were collected 2-4 days post-emergence and were injected with Dengue type 2 virus NGC strain,while control group were injected with equal volume of physiological saline.Total RNA was isolated,and small RNAs ranging up to 30 nt in length were excised and analyzed using Illumina HiSeq 2000.After removing adaptor sequences and contaminated reads,Clean reads were aligned to Dengue virus genome and its complementary sequence by SOAP2.The length,strand ratio nucleotide bias and genome distribution of Dengue virus-derived vsRNAs and vpiRNAs were further analyzed.Results Total,3835 vsRNAs and 395 vpiRNAs unique tags were obtained,among them 94.99％ vpiRNAs were derived from virus genome strand,and distributed across the viral genome,but with an enrichment at several "hot-spots".Except weak bias for adenine at position 10 (10A)in the sense molecules as the feature of secondary piRNA,other signature piRNA characteristics were not observed,including a preference for a uridine at their 5'-end,which were main characteristics of primary piRNAs,and a significant 10 nt overlap (Ping-Pong)between sense and antisense piRNA.Conclusion Dengue virusderived piRNAs were indentified in Dengue virus infected Ae.albopictus.

ObjectiveTo investigated the effects of Rhodiola Compound on improving the intellective function in mice and provide the basis for clinical application.MethodsMice were divided to different groups of three doses of rhodiola compound (0.3 g/kg,0.6 g/kg,1.2 g/kg) and swimming abilities were tested.Other mice were administrated single dose of compound rhodiola( 1.2 g/kg) and training by Morris water maze.Drug's improving intelligence function was assessed using memory acquisition impaired models made by scopolamine or alcohol.When the Morris water maze test was finished,mice were killed and brains were removed immediately to measure SOD and NO levels.ResultsGroups of three doses of compound rhodiola could significantly prolong the swimming time(P ＜ 0.05,P ＜ 0.01 ).Compound Rhodiola group can significantly reduce the swimming distance than the untreated group( ethanol model group:(26 906.6 ± 2769.7 ) mm,RCE treated group:( 19 586.1 ± 6826.7 ) mm ; P ＜0.05 ).Swimming distance and time of cross-platform quadrant was significantly increased,comparing with model group (P ＜ 0.05 ).Compound Rhodiola significantly enhanced the activity of mouse brain's SOD ( Scopolamine model group:( 150.3 ± 17.7 ) U/ml,RCE treated group:( 197.9 ± 16.8 ) U/ml ; P ＜ 0.05 ) and NO levels ( Scopolamine model group:( 44.7 ± 16.7 ) μmol/gprot,RCE treated group:( 65.4 ± 14.5 ) μmol/gprot ; P ＜ 0.05 ) significantly.ConclusionCompound Rhodiola could promote mice learning and memory function,SOD and NO in brain maybe play a important role in this effect.

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.

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.

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.

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.

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.

Objective To explore the clinical features of Shenzhen service worker with upper gastrointestinal bleeding and association with medical insurance. Methods 169 patients(service workers in Shenzhen city)with upper gastrointestinal bleeding were retospectiveng analyzed, the treatment time, the severity of bleeding, the cause of the bleeding and bleeding incentive, diagnosis and treatment compliance, and other characteristics were analyzed; the differences between the case of group health insurance and the case of group no-health insurance, were observedResults For the case of non-insurance group the treatment time was delayed( P＜0. 01 ) ;the non-standard pre-hospital diagnosis and treatment was higher( P＜0. 01 ). The rate of moderate and severe cases was higher( P＜0. 05 ). The rate of the patients do not comply with gastroscopy and refused hospitalization, the non-insurance group was higher than the insurance group(P＜0. 01 and P＜0. 05 respectively). In compliance with the treatment of the cases,ly the efficiency and the total effective rate were sigmficantly higher in the insurance group than non-insurance. Conclusion Public participation in health insurance services enable the upper gastrointestinal bleeding in patients with timely medical treatment fixed medical institutions in order to get more early diagnosis and treatment,access to better efficacy and prognosis.