Objective To find the expression level and the role of ENST00000460164 in luminal A breast cancer.Methods The expression level of ENST00000460164 in breast cancer tissues was detected by RT-qPCR.pll3.7-ENST00000460164-shRNA and empty vector,pll3.7,were transfected into MCF-7 cells.Cell cycle was measured by flow cytometry.Western blot was used to detect the expression of P16INK4A and cyclinD1.Results ENST00000460164 was highly expressed in luminal A breast cancer tissues as compared to the adjacent non-cancer tissues.The knockdown of ENST00000460164 resulted in the G1 cell-cycle arrested,cyclin D1 downregulated and P16INK4A upregulated in MCF-7 cells.Conclusions ENST00000460164 is overexpressed in luminal A breast cancer.ENST00000460164 may control G1/S transition by regulating P16INK4A or cyclin D1 expression.

Objective To evaluate the relationship between cold hyperalgesia and trafficking of transient receptor potential melastatin 8 (TRPM8) to cell membrane in the dorsal root ganglion (DRG) of rats with neuropathic pain (NP).Methods Ninety-six healthy male Sprague-Dawley rats,aged 10-12 weeks,weighing 250-280 g,were divided into sham operation group (S group,n=48) and NP group (n =48) using a random number table.NP was produced by chronic constriction injury to the sciatic nerve.The number of paw lifts on the cold plate and mechanical paw withdrawal threshold (MWT) were measured on 1 day before operation and 1,4,7,10 and 14 days after operation.Rats were sacrificed after behavioral testing,and ipsilateral DRGs of the lumbar segment (L46) were dissected tor detection of the expression of TRPM8 in total and membrane proteins by Western blot,and the ratio of TRPM8 expression in the membrane protein to that in the total protein (m/t ratio) was calculated.Results Compared with group S,the number of paw lifts on the cold plate was significantly increased,the MWT was decreased,the expression of TRPM8 in total and membrane proteins was up-regulated,and m/t ratio was increased on postoperative days 4,7,10 and 14 in group NP (P＜0.05 or 0.01).In group NP,the number of paw lifts on the cold plate was gradually increased with the prolongation of time after operation and reached the peak on postoperative day 10,maintaining at the peak until postoperative day 14;the MWT was gradually decreased and reached the lowest level on postoperative day 10,maintaining at the lowest level until postoperative day 14;the expression of TRPM8 in total and membrane proteins and m/t ratio were gradually increased with the prolongation of time after operation and reached the peak on postoperative day 10,maintaining at the peak level until postoperative day 14 (P＜0.01).Conclusion The mechanism underlying the development of cold hyperalgesia is related to enhanced trafficking of TRPM8 to cell membrane in DRGs of rats with NP.

Objective To understand the present situation of safety organizing and quality of nurse work life in pediatric intensive care unit(PICU),and to discuss the relationship between safety organizing and quality of nurse work life.Methods A total of 279 PICU nurses in eight hospitals in Zhejiang were selected by the safety organizing scale (SOS)and quality of nursing work life(QNWL).Results The score of SOS was (45.23 ±10.88)points,and the score of QNWL was (159.46 ±33.60)points.There was a positive correlation between safety organizing and quality of nurse work life(r =0.256,P <0.05).Conclusion The nurse managers should improve the level of quality of nurse work life by safety organizing.

patients received thoracic surgery. Methods Fifty patients underwent radical resection of pulmonary carcinoma were divided into DP group ( dexmedetomidine infusion by micro pumo and parecoxib 40 mg through intravenous injection) and DN group ( dexmedetomidine infusion by micro pumo and nomal saline 5mL through intravenous injection). The time for spontaneous breath,eye opening and extubation after the oper-ation were recorded. The mimi-mental state examination ( MMSE) was used to assess the cognitive function. The concentration of protein S100β and IL-6,TNF-α were determined at 1 d before operation and 1 h,24 h after the operation. Results The difference of time for spon-taneous breath,eye opening,extubation and preoperative values of MMSE between two groups were not statistically significant(P>0. 05). MMSE scores at 1 d,3 d and 5 d after operation in group DN were lower than those in group DP (P<0. 05). The values of S100β、IL-6、TNF-α at 1 h and 24 h after operation in group DN were higher than that in group DP (P< 0. 05). Conclusion Dexmedetomidine com-bined with parecoxib can decrease the incidence of postoperative cognitive dysfunction in elderly patients underwent thoracic surgery,and the mechanism of which may be related to the downregulation of serum inflammatory factors.

Objective To investigate the expression of monocyte-macrophage-related factors and interstitial fibrosis in kidney tissues of rats with ureter obstruction and recanalization .Methods Forty-eight male Spragur-Dawley rats were divided randomly into the obstructive group:sham (n=6), unilateral ureteral obstruction(UUO)3 days (n=6), UUO 7 days (n=6), and UUO 14 days (n=6) and recanalization group:bilateral ureteral obstruction(RBUO)0 day (n=6), 3 days after RBUO (n=6), 7 days after RBUO (n=6), and 14 days after RBUO (n=6).The kidneys were excised on day 3, 7, and 14, and the deposition of collagen fibers in kidney was detected with HE and Masson staining . Immunohistochemical analysis was performed to evaluate the protein expressions of monocyte chemoattractant protein -1 (MCP-1), macrophage colony-stimulating factor (M-CSF) and activated-macrophage marker CD68.Real-time PCR was used to detect the mRNA expressions of MCP-1 and M-CSF.TGF-β1 levels were determined by ELISA .Results Fibrosis observed with HE and Masson staining was obviously increased in kidney tissue of UUO rats , and aggravated as time prolonged, but alleviated in rats with recanalization .TGF-β1 levels were increased obviously in the UUO group , but decreased in rats with recanalization compared with those in BUO rats .In UUO rats, mRNA and protein expression levels of MCP-1 and M-CSF were increased .MCP-1 and M-CSF expression was gradually decreased in rats with recanalization compared with those in BUO rats .The dynamic change in expression of MCP-1 and M-CSF in both UUO rats and recanalization rats was consistent with the change in expression of CD 68. Conclusion Dynamic change in expression of MCP-1 and M-CSF in kidney tissues reflects change of activated and accumulated monocyte -macrophages , which may be one of the major mechanisms contributing to fibrosis induced by ureter obstruction .Renal fibrosis is alleviated by down-regulated expression of monocyte-macrophages factors with recanalization operation .

Objective: To observe the effect of electroacupuncture (EA) at Zusanli (ST 36) on the gastrointestinal motility and the ultrastructures of pacemaker cells [the interstitial cells of Cajal (ICC)] in diabetic gastroparesis (DGP) rats and explore the mechanism of EA for DGP.Methods: A total of 50 Sprague-Dawley (SD) rats were randomly divided into group A, group B, group C, group D and group E, with 10 rats in each group. Group A was the blank control; a single intraperitoneal injection of 2% streptozotocin (STZ) was performed in rats of group B, group C, group D and group E, with high glucose and high fat diet for 8 weeks to establish the DGP rat models. Group B was the model group and the rats did not receive any treatment; group C was EA at acupoint group and the rats received EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6); group D was EA at non-acupoint group and the rats received EA at the control points of Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6); group E was metoclopramide group and the rats were treated by intragastric administration of metoclopramide. Blood glucose was detected using ONE TOUCH blood glucose meter; gastric emptying rate and small intestine migration rate were measured using intragastric phenol red; ultrastructures of gastric antrum ICC were detected by transmission electron microscopy.Results: The differences of blood glucose between group B, group C, group D, group E versus that of group A were statistically significant after modeling (P<0.01); after treatment, the differences of blood glucose between group D, group E versus that of group C were statistically significant (P<0.05,P<0.01); the gastric emptying rate of rats in group B was statistically significant different from that in group A (P<0.01); the gastric emptying rate of rats in group C was statistically significant different from that in group B (P<0.01). The migration rates of rats' small intestines in group B, group C, group D and group E were all statistically significant different from that in group A (P<0.01); the migration rate of rats' small intestines in group C was statistically significant different from that in group B (P<0.01). The ultrastructure of rat's ICC in group B showed apoptosis compared with that in group A; rat's ICC in group C had complete basement membrane, more cytoplasm mitochondria, Golgi and rough endoplasmic reticulum, showing clear structure, occasional mitochondria swelling and gap junctions with adjacent smooth muscle cells; there were no significant differences between group D, group E versus group B.Conclusion: EA at Zusanli (ST 36) plus other acupoints can regulate the blood glucose and promote gastrointestinal motility in DGP rats, and the mechanism may be related to repairing the damaged ICC structure.

AIM:To investigate the effect of cyclopamine on Hedgehog (HH) signaling, phenotypic transfor-mation and matrix accumulation induced by aristolochic acid (AA) in renal tubular epithelial cell NRK-52E.METHODS:NRK-52E cells were randomly divided into control group (treated with solvent only), AA group (treated with AA at con-centrations of 1, 5, 10 mg/L) and cyclopamine group (treated with AA at concentration of 10 mg/L plus cyclopamine at concentrations of 1, 5, 10μmol/L).After cultured for 24 h, the mRNA expression of Ptch1, Smo,α-SMA, E-cadherin, ZO-1, BMP-7, type I collagen and type III collagen was quantified by real-time PCR.The protein levels of Shh and TGF-β1 were detected by ELISA .Immunofluorescence staining was used to evaluate the expression of Ptch 1, Smo,α-SMA, E-cadherin and type III collagen in the NRK-52E cells.RESULTS: AA increased the expression of TGF-β1, α-SMA and type III collagen, decreased the expression of E-cadherin and ZO-1 protein, and down-regulated the expression of Ptch1, Shh and Smo mRNA in the NRK-52E cells, indicating that AA activated HH signaling , and phenotypic transformation and matrix accumulation occurred in AA-treated NRK-52E cells.Treatment with cyclopamine inhibited HH signaling by decrea-sing Smo expression and increasing Ptch 1 expression.Moreover, cyclopamine also down-regulated the expression of TGF-β1,α-SMA, type I collagen and III collagen , and up-regulated the expression of BMP-7, ZO-1 and E-cadherin.CON-CLUSION:AA induces phenotypic transformation and matrix accumulation in renal tubular epithelial cells , which can be inhibited by cyclopamine treatment .The possible mechanism is that cyclopamine suppresses the activation of HH signaling , resulting in the reduction of epithelial-to-mesenchymal transition and matrix deposition .

Objective Pneumosilicosis is characterized by pulmonary fibrosis and cannot be effectively treated at present. This study was to explore the changes of monocyte subsets in the mouse model of silicon dioxide-induced experimental pneumosilicosis and the correlation of the changes with lung inflammatory injury and pulmonary fibrosis. Methods A total of 100 male C57BL/6J mice weighing 18-22 g were equally randomized into a normal saline (NS) group and a silicon dioxide (quartz) group.The model of experimental pneumosilicosis was established by oropharyngeal aspiration of quartz suspension.At 1, 3, 7, 14, and 28 days after treat-ment, the mice were sacrificed and the proportions of different circulating monocyte subpopulations determined by flow cytometry.Dif-ferent types of inflammatory cells in the bronchoalveolar lavage fluid ( BALF) were routinely counted.The inflammation score and col-lagen volume fraction ( CVF) of the lung tissue were obtained by HE and picrosirius red staining. Results At 7 days after quartz treatment, silicotic nodules were observed in the lung tissue.Compared with the NS controls, the model mice showed significantly in-creased inflammation score and CVF at 7 days (0.920 ±0.049 vs 1.400 ±0.089, P<0.01;0.525 ±0.048 vs 1.950 ±0.065, P<0.01) and 28 days (0.800 ±0.089 vs 1.520 ±0.136, P<0.01;0.850 ±0.050 sv 5.300 ±0.776, P<0.01).In comparison with the NS group, the quartz group also exhibited significant increases in the number of total cells at days 1-28 (P<0.01) and the count of neutrophils at days 1-14 (P<0.01) in the bronchoalveolar lavage fluid (BALF) of the model mice, as well as in the number of macrophages in the BALF at 3 days (0.980 ±0.663 vs 6.821 ±2.627, P<0.01), 7 days (1.225 ±0.601 vs 6.697 ±1.864, P<0.01), 14 days (1.492 ±0.438 vs 2.574 ±0.396, P<0.01), and 28 days (2.035 ±0.456 vs 3.249 ±0.492, P<0.01).The count of neutrophilic granulocytes in the BALF was remarkably higher in the quartz than in the NS group at 1, 3, 7, and 14 days (P<0.01) but not at 28 days (P>0.05).Compared with the NS controls, the quartz-treated mice showed markedly increased proportion of Ly6Chimonocytes at all time points, which peaked at 7 days (58.750 ±2.386 vs 78.300 ±2.517, P<0.01), with a positive corre-lation with the inflammation score (P<0.01) and CVF of the lung tissue (P<0.01) at 7 and 28 day. Conclusion The propor-tions of circulating Ly6Chi and Ly6Clo monocytes changed dynamically in the murine model of quartz-induced experimental pneumosilico-sis.The increased proportion of the Ly6Chi monocyte subpopulation might be closely related with lung inflammatory injury and pulmona-ry fibrosis in pneumosilicosis.

Objective: To observe the effect of electroacupuncture (EA) on the electrogastrogram and gastric antrum ghrelin in rats with diabetic gastroparesis (DGP). Methods: Fifty Sprague-Dawley (SD) rats were randomly divided into group A, group B, group C, group D and group E, 10 rats in each group. Group A was the blank control group without intervention. Group B, Group C, Group D and Group E were treated with single dose intraperitoneal injection of 2% streptozotocin (STZ), combined with 8-week high glucose and high fat diet to establish DGP rat models. Group B was the model group without treatment. Group C was the EA at acupoint group, was treated with EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6). Group D was the EA at non-acupoint group, was treated by EA at the control points of Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6). Rats in the metoclopramide control group received 1.7% metoclopramide solution [10 mL/(kg·bw)] by gavage. Rat's blood glucose level was measured by blood glucose meter; gastric emptying rate was detected using phenol red as a marker; the electrogastrogram was detected by BL-420F biological function system; the protein level of ghrelin was detected by enzyme-linked immunosorbent assay (ELISA); the expression of ghrelin mRNA was detected by real-time polymerase chain reaction (RT-PCR). Results: Compared with group A, the blood glucose of group B, C, D and E were significantly increased before and after the treatment (all P<0.01); after treatment, the gastric emptying rate of group B was significantly decreased (P<0.01), the migration rates of small intestine in group B, C, D and E were significantly lower (all P<0.01), and the protein content of ghrelin in group C was significantly decreased (P<0.01); the expressions of ghrelin mRNA were significantly increased in group B, C, D and E (all P<0.01), the mean amplitudes of electrogastrogram in group B and D were significantly decreased (both P<0.01). After treatment, compared with group B, the blood glucose of group C was significantly decreased (P<0.05), the gastric emptying rate and small intestine migration rate were significantly increased in group C and E (P<0.05, P<0.01), the small intestinal migration rate was significantly increased in group D (P<0.05), the expression of ghrelin in protein and mRNA in group C was significantly lower (P<0.01), the expression of ghrelin mRNA in group E was significantly lower (P<0.05), and the mean amplitude of electrogastrogram in group C was significantly increased (P<0.05). After treatment, compared with group D, the protein and mRNA expressions of ghrelin in group C were significantly decreased (P<0.01). After treatment, compared with group E, the protein expression of ghrelin in group C was significantly decreased (P<0.01). Conclusion: EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6) could regulate the blood glucose level of DGP model rats, enhance electrogastrogram activity, promote gastric emptying, and regulate ghrelin expression in protein and mRNA.

AIM: To establish a cell line of stable silencing of P2X7 receptor (P2X7R) expression through short hairpin RNA ( shRNA)-mediated interference in murine RAW264.7 macrophages, and to investigate the proliferation and apoptosis in the cell line.METHODS:Stable silencing of P2X7 R gene in the RAW264.7 cells was achieved by re-combinant shRNA plasmid targeting murine P2X7 R gene via liposome mediated transfection, followed by G418 selection. The efficacy of plasmid transfection and P2X7 R silencing in G418 resistant cells was verified by immunofluorescent micros-copy and real-time PCR, respectively.The proliferative activity was analyzed by CCK-8 assay and EdU cell proliferation as-say.The cell cycle distribution and apoptosis were evaluated by flow cytometry.RESULTS:The expression of P2X7 R at mRNA and protein levels was down-regulated by 80% in shP2X7 R group compared with negative control ( NC) plasmid transfection.In addition, P2X7 R-silencing cells exhibited higher proliferative activity compared with NC and wild-type RAW264.7 cells (P<0.05).Compared with NC cells, P2X7R silencing resulted in an increase in the phagocytosis of the cells ( P<0.05) .CONCLUSION:A cell line RAW264.7 of stable silencing of P2X7 R expression was successfully es-tablished.P2X7 R gene silencing stimulates the proliferation, and changes phagocytic function in murine RAW264.7 macro-phages.