Objective To evaluate the role of microglial activation in dorsal root ganglia in a rat model of persistent postoperative pain evoked by skin/muscle incision and retraction (SMIR).Methods Seventy male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 2 groups (n =35 each):group sham operation (group S) and group SMIR.The rat model of persistent postoperative pain evoked by SMIR was established according to the method described by Flatters.Pain behavior was assessed by mechanical paw withdrawal threshold to yon Frey filament stimulation at 1 day before and 1,3,7,12,22 and 32 days after operation.Five animals were sacrificed at each time point in each group for microglia count in dorsal root ganglia.Results Compared with group S,mechanical paw withdrawal threshold was significantly decreased at 3-22 days after operation,and microglia count was significantly increased at 3-12 days after operation in group SMIR (P ＜ 0.05).Conclusion Microglial activation in dorsal root ganglia may be involved in the development of SMIR-evoked persistent postoperative pain in rats.

Objective To evaluate the different effects of laryngeal mask and mask ventilation during bronchoscopy in elder patients. Methods 120 old patients (ASA I-III) were divided into three group according to the table of random number as following:surface anesthesia with autonomous respiration (group I);endoscope mask ventilation (group II);laryngeal mask airway ventilation (group III). Oxygen inhalation through nasal tube in groupⅠ,group II and group III was ventilated with endoscope mask and LMA respectively ,thenwe observed whether there were adverse effects or not during the procedure. Results The adverse reactions of group I were more than other groups. Compared with the group I, variance of MAP,HR, RR, SpO2 in the groupⅡand groupⅢ had less changes, the cases with severe bucking decreased significantly, and satisfaction degree increased markedly, (P <0.01or P<0.05 ) . The incidence rate of resp iratory depression and airway obstruction in groupⅡwas higher than that in groupⅢ (P<0.05). Conclusion Endoscope mask ventilation and LMA can both be adopted in analgesia bronchoscopy for old patient. LMA under general anesthesia to transbronchial lung biopsy would control respiration according to the demand at any time.LMA has more advantage in the operation for the stable respiration and hemodynamics and less complications.

A biosensor was constructed to detect the endotoxin.The amine-terminated aptamer was immobilized on the gold electrode surface covered in advance with a self-assembled monolayer (SAM) of 3-mercaptopropionic acid (MPA).The modification of the gold electrode was confirmed by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS).It was found that MPA assembly time at 6 h in the gold electrode surface to form a stable self-assembled monolayer.The biosensor has a good linear relationship with concentration of endotoxin in the range of 0.001 ～0.1 EU/ml,it is possible to be applied for quality control in the actual biological sample.

Objective To investigate the effects of skin/muscle incision and retraction(SMIR)on mechanical paw withdrawal threshold and the ability of spatial learning and memory in immature rats after adulthood. Methods 27 male SD rats aged 3 weeks and weighing 60 ~ 80 g were randomly divided into 3 groups(n = 9):control group(group C),sham operation group(group Sham)and skin/muscle incision and retraction group (group SMIR). Group SMIR received operation for skin/muscle incision and retraction. Sham group received skin/muscle incision but no retraction.No surgery was operated on C group. Pain behavior was assessed by mechanical paw withdrawal threshold(MWT)to von Frey filament stimulation before and 1,3,7,12,22 and 32 days after operation.The effects of spatial learning and memory function were assessed by Morris water-maze test at 33 days after operation. Results Mechanical paw withdrawal threshold of group SMIR decreased 1 day after operation (P<0.05)and showed no significant difference before and 3,7,12,22,32 days after operation in 3 groups(P >0.05). In Morris water-maze test,compared with Sham and C group,the average escape latency in SMIR was sig-nificantly longer in the water maze navigation experiment(P < 0.01);the ratios of time and path in the quadrant of the platform were obviously lower in SMIR(P < 0.01). There was no statistical difference between sham and C group(P>0.05).Conclusion SMIR did not cause chronic pain but may cause a decrease in the ability of spatial learning and memory in immature rats.

Due to the complexity of bioactive ingredients in biological samples,the screening of target proteins is a complex process.Herein,a feasible strategy for directing protein immobilization on silica magnetic beads for ligand fishing based on SpyTag/SpyCatcher(ST/SC)-mediated anchoring is presented.Carboxyl functional groups on the surface of silica-coated magnetic beads(SMBs)were coupled with SC using the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysulfosuccinimide method,named SC-SMBs.The green fluorescent protein(GFP),as the capturing protein model,was ST-labeled and anchored at a specific orientation onto the surface of SC-SMBs directly from relevant cell lysates via ST/SC self-ligation.The characteristics of the SC-SMBs were studied via electron microscopy,energy dispersive spectroscopy,and Fourier transform infrared spectroscopy.The spontaneity and site-specificity of this unique reaction were confirmed via electrophoresis and fluorescence analyses.Although the alkaline stability of ST-GFP-ligated SC-SMBs was not ideal,the formed isopeptide bond was unbreakable under acidic conditions(0.05 M glycine-HCl buffer,pH 1-6)for 2 h,under 20％ethanol solution within 7 days,and at most temperatures.We,therefore,present a simple and universal strategy for the preparation of diverse protein-functionalized SMBs for ligand fishing,prompting its usage on drug screening and target finding.

Objective:To investigate the efficacy and mechanism of canagliflozin (Cana) in the treatment of high glucose-induced human podocyte (HPC) injury.Methods:The HPCs were divided into 5 groups: normal glucose group (NG group), mannitol group (MA group), high glucose group (HG group), Cana low dose (0.3 μmol/L) group and Cana high dose (1.0 μmol/L) group. Western blotting was used to examine the protein expressions of membrane-associated guanylate kinase inverted-2 (MAGI2), podocyte-associated protein nephrin, sodium-glucose transporter 2 (SGLT2), NOD-like receptor thermal protein domain associated protein 3 (NLRP3), apoptosis- associated speck-like protein containing a CARD (ASC), and cleaved-caspase1 in podocytes. Phalloidin staining of F-actin in podocytes was used to observe cytoskeletal injury. Intracellular reactive oxygen species (ROS) level of HPC was detected by the 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) probe. Levels of interleukin (IL)-18 and IL-1β in culture medium of podocytes were detected by enzyme-linked immunosorbent assay (ELISA).Results:(1) Compared with the NG group, the protein expressions of MAGI2 and nephrin decreased (both P<0.01), the protein expression of SGLT2 increased ( P<0.01), the changes of cell morphology and cytoskeleton remodeling were obvious, intracellular ROS level increased ( P<0.01), while NLRP3, ASC and cleaved-caspase1 protein expressions decreased in the HG group (all P<0.01). The results of ELISA showed that IL-18 and IL-1β concentrations were higher in the HG group (both P<0.05). (2) Compared with the HG group, in the Cana groups, MAGI2 and nephrin expressions up-regulated (both P<0.01), the changes of cell morphology and cytoskeleton remodeling were alleviated. Meanwhile the Cana groups showed decreased SGLT2 expression ( P<0.05), lower ROS level, down- regulated NLRP3, ASC, cleaved-caspase1 expressions (all P<0.01), and decreased concentrations of IL-18 and IL-1β in culture medium of podocytes (both P<0.05). Conclusion:Cana can improve high glucose-induced injury and inflammation in human podocyte, possibly due to the repression of the ROS/NLRP3 signaling pathway.