Human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) might be developed as a novel anti-tumor drug due to its selective cytotoxicity in tumor cells. The predicted Macaca mulatta TRAIL (mmTRAIL) is highly homologous to hTRAIL in nucleotide acid as well as amino acid sequence, suggesting that mmTRAIL might induce apoptosis of human cancer cells. However, the cytotoxicity of mmTRAIL in human cancer cells has not been investigated. In this paper, it is reported that the gene encoding mmTRAIL has been cloned by using reverse-transcriptase polymerase chain reaction (RT-PCR) from monkey peripheral blood mononuclear cells (PBMCs) in our laboratory. Subsequently, an expression plasmid was constructed by inserting mmTRAIL gene into pQE30 plasmid. After induction by addition of Isopropyl β-D-1-Thiogalactopyranoside (IPTG), mmTRAIL was expressed. MmTRAIL was recovered from supernatant of sonicated bacteria by Ni-NTA agarose affinity chromatography. SDS-PAGE and gel filtration chromatography demonstrated that mmTRAIL forms trimer in solution. In vitro assays indicated that mmTRAIL was cytotoxic to human COLO205 tumor cells but not to normal cells at low concentration of nanomole. In addition, antitumor effect of mmTRAIL was evaluated in mice bearing human COLO205 tumor xenografts. Intratumorally injected mmTRAIL significantly inhibited growth of tumor grafts. These results suggested that mmTRAIL was valuable as candidate drug for cancer-targeted therapy.
Animals ; Antineoplastic Agents ; Apoptosis ; Cell Line, Tumor ; Cloning, Molecular ; Humans ; Leukocytes, Mononuclear ; Macaca mulatta ; Mice ; Plasmids ; TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; Xenograft Model Antitumor Assays

Transcriptional regulation is one of the major regulations in plant adventious shoot regeneration, but the exact mechanism remains unclear. In our study, the RNA-seq technology based on the IlluminaHiSeq 2000 sequencing platform was used to identify differentially expressed transcription factor (TF) encoding genes during callus formation stage and adventious shoot regeneration stage between wild type and adventious shoot formation defective mutant be1-3 and during the transition from dedifferentiation to redifferentiation stage in wildtype WS. Results show that 155 TFs were differentially expressed between be1-3 mutant and wild type during callus formation, of which 97 genes were up-regulated, and 58 genes were down-regulated; and that 68 genes were differentially expressed during redifferentiation stage, with 40 genes up-regulated and 28 genes down-regulated; whereas at the transition stage from dedifferentiation to redifferention in WS wild type explants, a total of 231 differentially expressed TF genes were identified, including 160 up-regualted genes and 71 down-regulated genes. Among these TF genes, the adventious shoot related transcription factor 1 (ART1) gene encoding a MYB-related (v-myb avian myeloblastosis viral oncogene homolog) TF, was up-regulated 3 217 folds, and was the highest up-regulated gene during be1-3 callus formation. Over expression of the ART1 gene caused defects in callus formation and shoot regeneration and inhibited seedling growth, indicating that the ART1 gene is a negative regulator of callus formation and shoot regeneration. This work not only enriches our knowledge about the transcriptional regulation mechanism of adventious shoot regeneration, but also provides valuable information on candidate TF genes associated with adventious shoot regeneration for future research.
Arabidopsis ; growth & development ; Arabidopsis Proteins ; physiology ; Gene Expression Regulation, Plant ; Genes, Plant ; Plant Shoots ; growth & development ; RNA ; Regeneration ; Seedlings ; growth & development ; Transcription Factors ; physiology ; Up-Regulation

Objective To study the role of ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil in neovascularization of human high malignant breast cancer .Methods Human high malignant breast cancer cell MDA-MB-231 and tumor-bearing nude mice established with MDA-MB-231 were treated with different doses of ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil .Epidermal growth factor receptor ( EGFR) expression level was examined by Western blotting and the RNA expression levels of neovascularization related molecules such as EGFR , vascular endothelial growth factor (VEGF), metalloproteinases(MMPs), thrombospondin(TSP-1), platelet derived growth factor( PDGF) , fibroblast growth factor ( FGF) were detected by Real-time PCR.Results Both ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil inhibited the expression of EGFR in vitro and in vivo in a dose-dependent way.Both compounds induced down-regulation of VEGF and up-regulation of TSP-1 at RNA level.The effect of Ganoderma lucidum extraction spore oil was more significant than that of ganoderma lucidum spore oil .Conclusion Both ganoderma lucidum spore oil and ganoderma lucidum extraction spore oil inhibite the expression of neovascularization related molecules and increase the expression of molecules inhibiting neovascularization , whereas the effect of ganoderma lucidum extraction spore oil is more obvious .

Objective To systematically evaluate the diagnostic value of magnetic resonance cholangiopancreatography (MRCP) in the investigation of bile duct anatomy of liver transplantation living donors.Methods A search in Cochrane library,MEDLINE,EMBASE,CBMdisc (China Biology Medicine disc) was performed to identify relevant English and Chinese-language abstracts,supplemented by Springer,OVID,Sciencedirect full text database,etc.Criteria for inclusion were based on validity criteria for diagnostic research published by the Cochrane collaboration.With Meta analysis package for Stata10.1,heterogeneity of the included articles was tested,which was used to select proper effect model to calculate pooled weighted sensitivity and specificity,positive likelihood ratio,negative likelihood ratio. Summary receiver operating characteristic (SROC) curve was performed and the area under the curve (AUC) was calculated. Finally,sensitivity analysis was performed.Results Seventeen articles with 34 studies were included.Heterogeneity analysis revealed heterogeneity between studies and the source was MRCP imaging methods spotted by meta-regression analysis. Subgroup analysis according to MRCP imaging methods showed homogeneity within subgroups.The pooled sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,diagnostic odd ratio of breath-holding thick slice MRCP,3D MRCP,the combination of the prior two methods,contrast enhance MRCP were 0.89,0.78,4.1,0.14,29; 0.92,0.80,4.5,0.10,45;0.95,0.82,5.2,0.06,85; and 1.00,0.76,4.1,0,1228,respectively with fixed effect model analysis.The area under the SROC curve was 0.83,0.92,0.96 and 0.99 respectively.Conclusion The combination of thick slice and 3D MRCP is a practical and effective method with good sensitivity and specificity to investigate bile duct anatomy of living liver transplantation donors,which fully meets the requirements of the preoperative assessment of bile duct structure.

Objective To investigate the effect of mild hypothermia combined with mitochondrial divison inhibitor 1 in mitochondrial after cerebral ischemia-reperfusion (IR).Methods Fourty male healthy Sprague-Dawley (SD) rats, weighing 280-320 g, were randomly divided into 5 groups (n=8 each): group Sham, group IR, hypothermia group (group H), Mdivi-1 group (group M) and hypothermia+Mdivi-1 group (group HM).Animal models of global cerebral IR were established by transoesophageal cardiac pacing inducing cardiac arrest followed by cardiopulmonary resuscitation (ischemia 4 min and reperfusion 6 h).The group Sham was similarly treated to group IR except the cardiac arrest and cardiopulmonary resuscitation.In groups H and HM, the core temperature was cooled down to 32-34℃ within 15 min starting from the beginning of reperfusion, and maintained for 6 h.In the other groups, the core temperature was maintained at the normal temperature.In groups M and HM, the animals were given Mdivi-1 (1.2 mg/kg) intravenously at the beginning of the reperfusion and the other groups were given the same Volume of dimethylsnlfone (DMSO).After 6 h of reperfusion, the rats were sacrificed, and bilateral hippocampi were immediately removed for determination the protein level of dynamin-related proten 1 (Drp1) and cytochrome C (Cyt-C) expression by Western blot and obsevation of the mitochondrial structure of pyramidal cell in hippocampal CA1 under electronic microscope.Results Compared with group Sham, the expression of Drp1 and Cyt-C was up-regulated in groups IR, H, M and HM (P＜0.05).Compared with group IR, the expression of Drp1 and Cyt-C was down-regulated in groups H, M and HM (P＜0.05).Compared with groups H and M, the expression of Drp1 and Cyt-C was down-regulated in group HM (P＜0.05).There was no significant difference in the expression of Drp1 and Cyt-C between groups H and M.The mitochondria were rod-shaped with clear and sound structure in group Sham, while mitochondria showed various degree of fission, swollen structures, matrix deposit, vacuoles formation and cristae collapse in other groups.The changes of group HM were relatively slight.Conclusion Mild hypothermia combined with mitochondrial divison inhibitor 1 alleviate mitochondrial damage after global cerebral IR of rats.The combined effect is better than that of any individual application.

Objective To explore the curative effect of Uvb radiation treatment on inflammation of radioactive oral mucosa and summarize nursing points. Methods Seventy patients with oral mucositis after radiotherapy for head and neck cancer were randomly divided into experiment group and control group, 35 in each group: The experiment group was treated by shortwave ultraviolet intracavitary irradiation and the control group, mouthwash by silver, 3 times a day, observing two groups of therapeutic effect after 3 d. Results The total effective rate of the observation group was statistically better higher than that of control group ( P < 0 . 05 ) . Conclusion Short-wave ultraviolet radiation is effective in the treatment of inflammation of radioactive oral cavity mucous membrane.

Objective To evaluate the effect of dexmedetomidine and tramadol on perioperative insulin resistance in patients undergoing radical resection of rectal carcinoma.Methods Sixty ASA I or II patients undergo-ing radical resection of rectal carcinoma were randomly divided into 3 groups(n =20 each):dexmedetomidine group (group D),tramadol group(group T),control group(group C).Group D was given dexmedetomidine intravenously at 1μg/kg 15min before induction of anesthesia followed by a continuous infusion of 0.5μg·kg -1 ·h -1 until the abdo-men was closed,and group T was given tramadol intravenously at 1.5mg/kg 15min before induction of anesthesia fol-lowed by a continuous infusion of 0.5mg·kg -1 ·h -1 until the abdomen was closed,whereas group C received the same volume of normal saline.Venous blood samples were taken at 30min before anesthesia induction(T1 ),1 h after the beginning of the operation(T2 ),1h after operation(T3 ),24h after operation(T4 )for determination of blood con-centrations of glucose(BG),insulin(INS),interleukin -6 (IL -6),tumor necrosis factor -α(TNF -α).Insulin resistance(HOMA -IR)and insulin sensitivity index(QUICKI)were calculated.The numbers of patients with PONV were studied respectively.Results The serum IL -6,TNF -α,BG,INS concentrations and HOMA -IR were signifi-cantly lower while ISI was significantly higher in both group D[t =7.71,3.37,8.78,8.73,11.45,2.82(T2 ),3.04, 2.95,12.75,10.73,16.09,2.92(T3 ),11.26,2.45,11.40,5.10,14.5,2.51(T4 ),all P <0.05]and group T[t =3.02,2.59,2.93,7.76,6.32,2.03(T2 ),8.78,2.27,4.14,8.83,7.68,2.12(T3 ),6.10,2.05,3.71,2.35,7.12, 2.09(T4 ),all P <0.05]at T2 ,T3 and T4 than those in group C.The serum TNF -αconcentration and HOMA -IR were significantly lower while ISI was significantly higher in group D[t =6.68,4.58,2.05 (T2 ),9.01,6.66,2.23 (T3 ),7.54,5.5,2.02(T4 ),all P <0.05]at T2 ,T3 and T4 than those in group T.The numbers of patients with PONV were significantly higher in group T than those in group D and group C (χ2 =26.13,18.75,all P <0.05 ). Conclusion Both dexmedetomidine and tramadol can attenuate perioperative insulin resistance in patients undergo-ing Radical Resection of Rectal Carcinoma,and the decrease the consentrations of IL -6 and TNF -αmay be involved in the mechanism.The roles of prevention of perioperative insulin resistance in dexmedetomidine group are superior to tramadol group.The incidence of PONV is less in a dexmedetomidine group than that in a tramadol group.

Objective To evaluate the effect of hypothermia on the expression of dynamin?related protein 1 ( Drp1) in brain tissues during global cerebral ischemia?reperfusion ( I∕R) in rats. Methods Thirty?six healthy male Sprague?Dawley rats, weighing 280-320 g, were divided into 3 groups ( n=12 each) using a random number table: sham operation group ( group Sham ) , global cerebral I∕R group ( group I∕R) and hypothermia group ( group H) . Cardiac arrest was induced by transoesophageal cardiac pacing followed by cardiopulmonary resuscitation to establish the global cerebral I∕R model in anesthetized rats in I∕R and H groups. In group H, the body temperature ( rectal temperature) was cooled down to 32-34 ℃ within 15 min starting from the beginning of reperfusion, and maintained at this level for 6 h. At 72 h of reperfusion, neurological deficit was scored, and the rats were sacrificed, and the whole brain was removed for examination of the pathological changes in hippocampal CA1 region and for determination of nor?mal pyramidal cell count and neuronal apoptosis in hippocampal CA1 region and expression of Drp1 and cy?tochrome c (Cyt c) in hippocampal tissues (by Western blot). The apoptosis rate was calculated. Re?sults Compared with group S, the neurological deficit score and apoptosis rate were significantly in?creased, and the number of normal pyramidal cells was decreased in I∕R and H groups, the expression of Drp1 and Cyt c in hippocampal tissues was significantly up?regulated in group I∕R ( P<0.05) , and no sig?nificant change was found in the expression of Drp1 and Cyt c in hippocampal tissues in group H ( P>0.05) . Compared with group I∕R, the neurological deficit score and apoptosis rate were significantly de?creased, the number of normal pyramidal cells was increased, and the expression of Drp1 and Cyt c in hip?pocampal tissues was down?regulated in group H ( P<0.05) . Conclusion The mechanism by which hypo?thermia inhibits cell apoptosis during global cerebral I∕R may be related to down?regulation of Drp1 expres?sion in rats.

Objective To investigate the feasibility of magnetic resonance diffusion tensor imaging (DTI) as a noninvasive way for assessment of kidney transplant function in the early posttransplantation period.Method Fifty-one kidney transplant recipients less than 1 month after kidney transplantation and 26 age-matched healthy volunteers were included and examined using a fatsaturated echo-planar DTI sequence in oblique-coronal orientation at 3.0 Tesla magnetic resonance (MR) imager (diffusion directions=6,b =0,300 s/mm2).According to the estimated glomerular filtration rate (eGFR) recorded and calculated on the examined day,all subjects were divided into four groups:group 1,healthy volunteers (n =26) ; group 2,eGFR≥60 mL/min/1.73 m2 (n =24) ; group 3,30≤eGFR＜60 mL/min/1.73 m2 (n =19) ; group 4,eGFR＜30 mL/min/1.73 m2 (n =8).Mean apparent diffusion coefficient (ADC) and mean fractional anisotropy (FA) were determined separately for the cortex and the medulla.The paired Students t test was used to compare ADC and FA between cortex and medulla within each group.ADC and FA between groups were compared by using the one-way analysis of variance test.Relationship between ADC and FA with eGFR of the transplants was assessed by using Pearson correlation analysis.Result Mean cortical FA was significantly higher in group 1 than in other three groups (P＜0.01 for all),while differences among groups of allograft recipients were not significant (P＞0.05 for all).There was a gradually decreasing trend of medullary FA and ADC,and cortical ADC from group 2 to group 4,and the differences among groups were all pronounced (P＜0.05 for all).In renal allografts,there was a significant positive correlation between eGFR and medullary FA,medullary ADC as well as cortical ADC (r =0.812,0.756,0.757,respectively,P＜0.01).The cortical-medullary discrimination of FA-map and ADC-map in group 3 and group 4 apparently decreased.Meanwhile,DTI revealed that the radial diffusion tracts in the medulla of group 3 and group 4 obviously broke off and reduced,especially in group 4.Conclusion DTI is a promising way to evaluate kidney transplant function early after transplantion,and can quantitatively and visually distinguish transplants with different functions.

Objective To explore the inhibitory effect of tetramethylpyrazine (TMP) on ultraviolet A-induced senescence as well as matrix metalloproteinase-1 (MMP-1) and-3 (MMP-3) mRNA expressions in human dermal fibroblasts (HDFs).Methods HDFs were isolated from the prepuce by enzymatic digestion, and subjected to primary culture.Cultured HDFs were randomly divided into several groups: control group cultured in high-glucose DMEM medium and receiving no treatment, three TMP groups treated with 20, 50 and 100 mg/L TMP respectively, UVA group receiving UVA radiation alone, UVA + TMP groups pretreated with 20, 50 and 100 mg/L TMP respectively for different durations followed by UVA radiation.UVA radiation was given once daily for 5 consecutive days.The 55th passage HDFs served as the P55 group (senescence control group).Subsequently, CCK-8 assay was performed to evaluate the proliferative activity of HDFs in vitro, optical microscopy to observe the morphologic changes of HDFs after UVA radiation, β-galactosidase staining to estimate the senescence in HDFs, and real-time fluorescence-based quantitative PCR to quantify the mRNA expressions of MMP-1 and MMP-3 in HDFs.Statistical analysis was carried out by one-way analysis of variance (ANOVA) followed by least significant difference (LSD)-t test or Dunnett's T3 test.Results Compared with the control group, the proliferation of HDFs was significantly but transiently inhibited in vitro after the treatment with 100 mg/L TMP for 48 hours (P ＜ 0.05), but showed no significant changes after the treatment with 20 or 50 mg/L TMP for 24, 48 or 72 hours or after the treatment with 100 mg/L TMP for 24 or 72 hours (all P ＜ 0.05).The pretreatments with TMP of 20, 50 and 100 mg/L for 24, 48 and 72 hours all promoted the proliferation of HDFs to a certain degree in the UVA + TMP groups compared with the UVA group, with significant differences in cellular proliferative activity among the UVA group, UVA + TMP groups and control group at 24, 48 and 72 hours (F =17.451,15.231, 23.535, all P ＜ 0.01).Compared with the UVA group, the proliferative activity of HDFs was significantly increased in UVA + 100-mg/L TMP group at 24, 48, 72 hours, UVA + 50-mg/L TMP group at 24 and 72 hours and UVA + 20-mg/L TMP group at 72 hours.After repetitive UVA radiation, HDFs in the UVA group experienced an increase in cell volume, granule acount, and β-galactosidase expression, which was similar to the changes in the P55 group, while the pretreatments with 20, 50 and 100 mg/L TMP for 24 hours suppressed these UVA-induced changes in HDFs.The percentage of β-galactosidase-positive HDFs was 68.417％ ± 1.181％ in the UVA group, 58.167％ ± 5.620％ in the UVA + 20-mg/L TMP group, 45.167％ ± 5.502％ in the UVA + 50-mg/L TMP group, 43.000％ ± 2.000％ in the UVA + 100-mg/L TMP group, 33.667％ ± 5.865％ in the control group, and 76.000％ ± 6.557％ in the P55 group, with significant differences among these groups (F =45.918, P ＜ 0.01).Furthermore, the UVA group significantly differed from the UVA + TMP groups and control group in the percentage of β-galactosidase-positive HDFs and mRNA expressions of MMP-1 and MMP-3 (all P ＜ 0.05).Conclusion TMP can protect HDFs against senescence induced by repetitive UVA radiation, and down-regulate the mRNA expressions of MMP-1 and MMP-3 during senescence.