AIM To investigate the changes of vascular systiolic/relaxant function in sino-aortic denervated rats. METHODS Male Sprague-Dawley rats were underwent sino-aortic denervation (SAD). The sinoaortic denervated (SAD) rats were adopted as a model of arterial baroreflex deficit. SAD, isolated aortic-denervated (AD) and isolated sinus-denervated (SD) rats were instrumented chronically to record blood pressure (BP), heart rate (HR), BP variability (BPV), HR variability (HRV), arterial baroreflex function control of heart period (ABR-HP) and BP (ABR-BP). The vascular maximum contractile/relaxant function was determined through cumulative venous injection of phenylephrine (SBP max ) and nitroprusside(DBP min ) both after ganglionic blokade. RESULTS Acute SAD(1 week after operation) caused hypertension and tachycardia in rats. Eighteen weeks after operation, BP and HR values in SAD and SD rats were not different from those in sham-operated rats, but AD rats were hypertensive compared with control group. Though the 24 h mean BP values of chronic (18 weeks after operation) SAD rats was not different from those in the sham-operated rats, 24 h BPV of SAD rats was significantly higher when compared with sham-operated rats. ABR function in the acute SAD rats was significantly decreased when compared with sham-operated rats, whereas in chronic SAD rats, both ABR-HP and ABR-BP were higher than those in acute SAD rats, but were still significantly lower than those in control groups. 18 weeks after operation, ABR function in SAD and AD rats were significantly decreased when compared with those in SD and control groups. SBP max after phenylephrine and DBP min after nitroprusside were significantly higher in SAD, AD and SD rats than in control group. ABR function was negatively correlated to DBP min ( r =-0.677 for ABR-HP, and r =-0.681 for ABR-BP; P

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.

AIM:To investigate the influence of matrine (MA) on the phenotype switching of mouse mono-cytes and alveolar macrophages induced by bleomycin ( BLM) .METHODS:All mice were randomly divided into normal saline (NS) group, BLM group, BLM+NS group and BLM +MA group.The mice were administered with BLM at 2.5 mg/kg via oropharyngeal instillation .The mice in BLM+MA group were treated with MA (15 mg· kg-1 · d-1 ) by oral gavage following BLM administration .The mice were sacrificed on days 3, 7, 14, and 21.The lungs were removed for pathological analysis .The circulating monocyte subsets and polarization state of bronchoalveolar lavage fluid ( BALF)-de-rived alveolar macrophages were analyzed by flow cytometry .RESULTS:The results of HE and Masson trichrome staining in BLM and BLM+NS groups exhibited classical pathological stages of lung fibrosis , including acute inflammation phase and later fibrosis phase .Compared with BLM +NS group, MA treatment alleviated the inflammatory response and the de-gree of fibrosis induced by BLM (P<0.05).There was a rapid change of circulating Ly6Chi monocytes and its magnitude was positively associated with the pulmonary inflammatory response .An expansion of M2-like alveolar macrophages was positively correlated with the magnitude of lung fibrosis .Moreover , MA treatment partially normalized the phenotype switc-hing of monocytes and alveolar macrophages .CONCLUSION:Matrine treatment attenuates BLM-induced pulmonary injury partially via modulating the phenotype switching of monocytes and alveolar mocrophages .

Objective To investigate the clinical characteristics and methods of diagnosis and treatment of granular lymphocytic leukemia (LGLL).Methods Clinical data of 3 patients with LGLL were retrospectively analyzed and relevant literature was reviewed.Results 3 patients were all onset with lymphocytosis,whose conditions progressed slowly.The diagnosis of 2 patients was T-LGLL with immunological characteristics of CD3+ CD4 CD8+ CD56-CD57+.The other patient' s diagnosis was NK-LGLL,whose immunological characteristic was CD3-CD4-CD8-CD56+ CD57-.Two of them didn' t need any treatment.One of them was treated with cyclosporine because of agranulocytosis and recurrent infection.Conclusions LGLL is a group of heterogeneous diseases,which clinical characteristic and prognosis are different.Flow cytometric immunopheotype,TCR Vβ analysis and TCR gene rearrangement are helpful to diagnosis.

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.

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 evaluate the changes in the expression of transient receptor potential vanilloid 1 (TRPV1) in dorsal root ganglions (DRGs) during remifentanil-induced hyperalgesia in the rats with incisional pain.Methods Thirty-two SPF healthy male Sprague-Dawley rats,weighing 240-260 g,aged 2-3 months,in which caudal catheters were successfully implanted,were divided into 4 groups (n=8 each) using a random number table:control group (group C),incisional pain group (group Ⅰ),remifentanil group (group R),and incisional pain + remifentanil group (group I+R).A 1 cm longitudinal incision was made through skin,fascia and muscle of the plantar aspect of the left hindpaw to establish the model of incisional pain.In group R,remifentanil was intravenously infused for 60 min at a rate of 1.2 μg · kg-1 · min-1.In group Ⅰ,the model of incisional pain was established,and the equal volume of normal saline was intravenously infused for 60 min at the same time.In group I+R,the model of incisional pain was established,and remifentanil was intravenously infused for 60 min at a rate of 1.2 μg · kg-1 · min-1 at the same time.In group C,the equal volume of normal saline was intravenously infused for 60 min.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawl latency (TWL) were measured at 24 h before normal saline or remifentanil infusion (To) and 2,6,24 and 48 h after the end of infusion (T1-4).The rats were sacrificed after the last measurement of pain threshold,and the DRGs of the lumbar segment (L4-6) were removed for determination of the expression of TRPV1 protein and mRNA by Western blot and real-time polymerase chain reaction,respectively.Results Compared with group C,the MWT was significantly decreased,and the TWL was shortened at T1-4,and the expression of TRPV1 protein and mRNA was up-regulated in R,I and I+R groups (P＜0.05).Compared with group R or group I,the MWT was significantly decreased,and the TWL was shortened at T1-4,and the expression of TRPV1 protein and mRNA was up-regulated in group I+R (P＜0.05) Conclusion Up-regulated expression of TRPV1 in DRGs may be involved in the mechanism underlying remifentanil-induced hyperalgesia in the rats with incisional pain.

Objective To study the hematopoietic stem cell injury(HSC)induced by busulfan. Methods C57BL/6 mice were treated with i.p. injection of 20 mg/kg or 40 mg/kg busulfan. All mice were euthanized at 15 days and 30 days after busulfan treatment for analysis of the peripheral blood cell counts, bone marrow cellularity and HPC (LKS-, lineage-sca1 -c-kit+), HSC(LSK, lineage-sca1 +c-kit+)and long-term HSC(CD34 - LSK, CD34 - lineage-sca1 +c-kit+)frequency. The colony-forming unit-granulocyte and macrophage(CFU-GM)ability of HPC was measured by colony-forming cell(CFC)assay,and the HSC self-renewal capacity was analyzed by single-cell colony-forming assay. Results The busulfan administration decreased the WBC,RBC and PLT compared with control mice. The HPC function (CFU-GM)was impaired(P < 0.05), and the HSC colony forming ability was decreased at 15 days after busulfan treatment(P < 0.05), whereas the body weight of the mice didn't change significantly after busulfan treatment. Conclusions Our findings suggest that busulfan can induce hematopoietic stem cell injury,and provide a support for the study of hematopoietic stem cell injury mechanism.

Objective:To put forward possible improvement measures through analyzing characteristics of pre-hospital first aid in Ali region and the problems confronted.Methods:The relevant data of pre-hospital first aid in the Emergency Department of Ali People's Hospital of Tibet from September 2015 to August 2019 were collected, and the epidemiological statistics were made. In addition, the number and cycle rule of pre-hospital first aid and the proportion of traffic injuries in pre-hospital first aid in different time periods were analyzed. The isochronous maps were drawn, and the residential areas, tourist attractions and main roads covered by the ground emergency medical service (GEMS) and helicopter emergency medical services (HEMS) at different periods were compared.Results:The epidemiology of pre-hospital first aid was mainly in middle-aged and young patients, accounting for 80.35%. Trauma patients were the most common in both non-native population (45.72%) and local residents, of which traffic injuries accounted for 66.43% of total trauma. Neurological emergency was the second most common among local residents (24.65%), and high altitude reaction was the second most common among non-native population (19.14%). From April to December every year, the amont of emergency treatment in Ali region increased periodically, and reached the peak from July to September. After eliminating the periodic influencing factors, the amount of pre-hospital first aid in Ali region showed a gradual increase over time. The regression equation was Y=15.7+0.27 X, F=36.809, P<0.05, R2=0.444. From April to December every year, the amount of pre-hospital first aid caused by traffic injuries increased significantly ( χ2= 10.819, P< 0.05). Within 15 min, GEMS could cover all the towns in Ali region, as well as the area where the first aid point was located and some villages that were pretty close to the first aid point, with a total of 49 villages (residences); Compared with 15 min, the pre-hospital resources could cover 75 villages (residences) and 4 tourist attractions within 1 h ( χ2 = 10.813, P < 0.05), and the main roads could cover about 788 km. If combined GEMS and HEMS, compared with the coverage of only one hour of ground emergency, the pre-hospital resources could cover 116 villages (residences) and 5 tourist attractions ( χ2 = 19.447, P < 0.05), and the main roads could cover about 1 234 km ( χ2 = 349.532, P < 0.05). Conclusions:HEMS combined with GEMS might fundamentally solve problem of comparatively small coverage of the emergency network in Ali region at present.