1.Inhibitory effect of IL-37 on inflammation-related diseases
Mengyuan LI ; Xingjiu YANG ; Ran GAO ; Damo XU
Chinese Journal of Comparative Medicine 2015;(12):75-80
Inteleukin-37 is a member of IL-1 family and originally named as IL-1F7.It has five different subtypes (IL-37a-e).It has been reported that IL-37 suppresses pro-inflammatory cytokines production of a variety of immune cells, and further regulate innate immune response. In addition, IL-37 has protective effects on colitis, arthritis, pancreatitis and other inflammatory diseases that induced by exogenous stimuli. In a word, IL-37 is a novel anti-inflammatory cytokine and plays an important role in a variety of inflammation-related diseases.
2.Expression, purification and activity assay of human IL-37b in E.coli
Mengyuan LI ; Rongfei WEI ; Damo XU ; Xingjiu YANG ; Ran GAO
Chinese Journal of Comparative Medicine 2017;27(3):20-24
Objective To investigate the expression of recombinant IL-37b protein and removal of the endotoxin, and identify its biological activity.Methods The prokaryotic expression vector pET28/IL-37b was constructed and to transform Escherichia coli (E.coli) Rosetta.After induction with IPTG, the recombinant protein was purified through Ni2+-NTA gel column and identified by SDS-PAGE and Coomassie brilliant blue staining.Then, the endotoxin protein was removed and was treated with LPS-stimulated RAW 264.7 cells.The culture supernatant was collected.The expression of IL-6 was detected by ELISA and the biological activity of the protein was identified.Results The recombinant IL-37b with high purity was expressed and the endotoxin produced by prokaryotic expression was reduced, and it was identified to have good biological activity.Conclusions In this study a recombinant IL-37b protein with high biological activity is successfully obtained.
3.Etablishment and phenotypic analysis of a C57BL/6 mouse model of S.Typhimurium infection
Rongfei WEI ; Mengyuan LI ; Damo XU ; Ran GAO
Chinese Journal of Comparative Medicine 2017;27(6):33-36,45
Objective To establish a C57BL/6 mouse model of intestinal infection induced by S.Typhimurium.Methods In order to improve the infectious sensitivity of S.Typhimurium, C57BL/6 mice were intragastrically given 5% (w/v) NaHCO3.Then mice were challenged with S.Typhimurium.The health condition, survival and body weight of mice were observed from day 0 to day 7 after the bacterial infection.The pathological changes were also examined.Results the mice challenged with S.Typhimurium showed decreased body weight and typical clinical signs, including in appetence, piloerection and low survival rate.Macroscopic dissection revealed that intestinal hyperemia and swelling were founded in the mice challenged with S.Typhimurium.Histopathology showed intestinal epithelial and mucosal damages.Conclusions We have successfully established a C57BL/6 mouse model of S.Typhimurium infection.This model may be of crucial significance for studying the biological functions of associated immunological molecules or cytokines in the process of inflammatory bowel disease induced by S.Typhimurium.
4.Effect of lung protective ventilation strategy on inflammatory responses in brain tissues of elderly patients requiring one-lung ventilation during radical resection for esophagus cancer
Wekhen GUO ; Mengyuan CHEN ; Tianfeng HUANG ; Yali GE ; Hong GAO ; Ju GAO
Chinese Journal of Anesthesiology 2017;37(2):139-142
Objective To evaluate the effect of lung protective ventilation strategy on inflammatory responses in brain tissues of elderly patients requiring one-lung ventilation (OLV) during radical resection for esophagus cancer.Methods Sixty patients of both sexes,aged 65-80 yr,weighing 45-75 kg,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,scheduled for elective radical resection for esophageal cancer,were divided into volume-controlled ventilation (VCV) group (n =30) and VCV plus protective ventilation strategy group (PV group,n =30) using a random number table.In group VCV,the tidal volume was set at 10 ml/kg during two-lung ventilation (TLV) and at 7 ml/kg during OLV with inspiratory/expiratory ratio 1:2.In group PV,the tidal volume was set at 7 ml/kg during TLV and at 5 ml/kg during OLV with inspiratory/expiratory ratio 1:2 and positive end-expiratory pressure 5 cmH2O,and lung recruitment maneuver was performed every 45 min.End-tidal pressure of carbon dioxide was maintained at 35-45 mmHg,and bispectral index value at 40-60 in both groups.Before induction of anesthesia (T1),at 10 min of TLV (T2),at 30 min of OLV (T3),at 15 min after restoration of TLV (T4) and at 24 h after operation (T5),jugular bulb venous blood samples were taken for determination of serum glial fibrillary acid protein,tumor necrosis factor-α and interleukin-6 concentrations by enzyme-linked immunosorbent assay.The cognitive function was assessed using Mini-Mental State Examination before operation (T0),at T5 and at 3 and 7 days after operation (T6,7).The occurrence of postoperative delirium was recorded.Results Compared with group VCV,the serum concentrations of tumor necrosis factor-α,interleukin-6 and glial fibrillary acid protein were significantly decreased at T3-5,Mini-Mental State Examination scores were increased at T6,7,and the incidence of postoperative delirium was decreased in group PV (P<0.05).Conclusion The mechanism by which lung protective ventilation strategy decreases the development of postoperative cerebral dysfunction is related to reduction of inflammatory responses in brain tissues of elderly patients requiring OLV during radical resection for esophagus cancer.
5.Establishment of two human tumor cell lines with lentivirus-mediated stably Smurf1 silencing and their effect on cell migration
Rongfei WEI ; Jing GUO ; Mengyuan LI ; Ruimin ZHU ; Xingjiu YANG ; Ran GAO
Chinese Journal of Comparative Medicine 2017;27(4):46-51
Objective To establish lentiviral expression vectors for Smurf1 silencing and assess the effects of Smurf1 silencing on cell migration.Methods HeLa and A549 cells were infected with lentiviral expression vectors for Smurf1 silencing respectively.After 7 days,the stable cell lines with Smurf1 silencing were obtained after puromycin-resistance screening,enrichment and expansion.The intracellular gene and protein levels of Smurf1 were detected by qPCR and western blot.Transwell assay was used to assess the effect of Smurf1 silencing on cell migration.Results The stable cell lines with Smurf1 silencing are constructed successfully.Silencing of Smurf1 down-regulated cell migration rate detected by Transwell assay.Conclusion Smurf1 promotes cell migration.
6.Effects of lung-protective ventilation on cerebral oxygen metabolism and postoperative cognitive function in elderly patients requiring one-lung ventilation
Weizhen GUO ; Mengyuan CHEN ; Ju GAO ; Yali GE ; Tianfeng HUANG ; Luojing ZHOU
Chinese Journal of Anesthesiology 2017;37(4):396-399
Objective To evaluate the effects of lung-protective ventilation on the cerebral oxygen metabolism and postoperative cognitive function in elderly patients requiring one-lung ventilation (OLV).Methods Sixty patients of both sexes,aged 65-80 yr,weighing 45-75 kg,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,scheduled for elective radical resection for esophageal cancer performed via video-assisted thoracoscope with general aneshesia,were divided into 2 groups (n =30 each)using a random number table:volume-controlled ventilation group (group VCV) and protective ventilation group (group PV).In group VCV,the tidal volume (VT) was set at 10 ml/kg during two-lung ventilation (TLV) and at 7 ml/kg during OLV.In group PV,the VT was set at 7 ml/kg during TLV and at 5 ml/kg during OLV with positive end-expiratory pressure of 5 cmH2O,and lung recruitment maneuver was performed every 45 min with inspiratory pressure at 15,20 and 25 cmH2O,PEEP 5 cmH2O,3 breaths per pressure,5 s/breath.Before induction of anesthesia (T1),at 10 min of TLV (T2),at 30 min of OLV (T3) and at 15 min after restoration of TLV (T4),blood samples were taken from the radial artery and jugular bulb for blood gas analysis,and pH value,arterial oxygen partial pressure (PaO2),arterial carbon dioxide partial pressure (PaCO2),arterial oxygen saturation (SaO2) and jugular venous oxygen saturation (SjvO2) were recorded.Oxygenation index (OI),intrapulmonary shunt (Qs/Qt),arteriovenous blood O2 content difference (Da-jvO2) and cerebral O2 extraction rate (CERO2) were calculated at the same time.Cognitive function was assessed using Mini-Mental State Examination at 7 days and 1 month after operation,and the development of postoperative cognitive dysfunction was recorded.Results PaO2,DajvO2,CERO2 and Qs/Qt were significantly higher and SjvO2 and OI were lower at T2-4 than at T1 in two groups (P<0.05).PaO2,SjvO2 and OI were significantly lower and Qs/Qt and CERO2 were higher at T3 than at T2,and Da-jvO2 was higher at T3-4 than at T2 in two groups (P<0.05).Compared with group VCV,PaO2,PaCO2,SjvO2 and OI were significantly increased and Qs/Qt,Da-jvO2 and CERO2 were decreased at T3,the Mini-Mental State Examination scores were increased on postoperative day 7,and the incidence of postoperative cognitive dysfunction was decreased in group PV (P<O.05).Conclusion Lungprotective ventilation is helpful in improving postoperative brain function of elderly patients requiring OLV.
7.Evaluation of ketamine-induced cerebral protection in mice with traumatic brain injury by magnetic resonance imaging
Xuan GAO ; Fang FANG ; Xiaomin LING ; Ruixue SONG ; Mengyuan PENG ; Zhanggang XUE ; Jing CANG
Chinese Journal of Anesthesiology 2017;37(4):501-503
Objective To evaluate ketamine-induced cerebral protection in mice with traumatic brain injury (TBI) by magnetic resonance imaging (MRI).Methods Thirty-two pathogen-free healthy male C57BL/6 mice,aged 8 weeks,weighing 26-30 g,were divided into 4 groups using a random number table:control group (group C,n=7),ketanine group (group K,n=7),TBI group (n=9) and TBI plus ketamine group (group TBI+K,n =9).TBI was produced with a pneumatically driven controlled cortical impact device.Ketamine 150 mg/kg was intraperitoneally injected at l h after operation in TBI+K and K groups,while the equal volume of normal saline was given instead in TBI and C groups.Open field test was conducted at 24 h,72 h and 7 days after operation.The animals in TBI and TBI+K groups were scanned by T1-weighted MRI at 6,24 and 72 h after operation,the animals in C and K groups were scanned by MRI at 24 h after operation,and the development of cerebral edema was observed.Results MRI scan showed no cerebral edema in C and K groups,and different degrees of cerebral edema were found in TBI and TBI+K groups.Compared with group C,the locomotor distance was significantly shortened at 24 and 72 h after operation in group TBI (P<0.05).Compared with group TBI,the size of cerebral edema was significantly decreased,and the locomotor distance was prolonged at 24 and 72 h after operation in group TBI+K (P<0.05 or 0.01).Conclusion MRI method further clarifies that ketamine can produce cerebral protection to some extent in mice with TBI.
8.Effect of Smad3 on cell migration of A549 and HeLa cells
Rongfei WEI ; Mengyuan LI ; Xingjiu YANG ; Ruimin ZHU ; Damo XU ; Ran GAO
Chinese Journal of Comparative Medicine 2017;27(1):11-15,21
Objective To investigate the effect of Smad3 on cell migration of A549 and HeLa cells.Methods Primers for pCMV-Myc-Smad3 plasmid construction and siRNA targeting Smad 3 were designed and synthesized .pCMV-Myc-Smad3 plasmid was constructed with molecular cloning techniques .Overexpression of Smad 3 with Myc-tag or silencing of endogenous Smad3, and then scratch assay was used to detect the migration ability of A 549 and HeLa cells in vitro. Results pCMV-Myc-Smad3 plasmid was successfully constructed .Overexpression of Smad 3 significantly up-regulated the migration rate of A549 and HeLa cells.Conversely, in the same cells, silencing of endogenous Smad3 or treatment with Smad3 inhibitor, SIS3, down-regulated the migration rate .Conclusions Smad3 promotes cell migration of A549 and HeLa cells.
9.Establishment and application of a patient derived xenograft mouse model of liver cancer
Hao HUANG ; Baoliang LI ; Xingjiu YANG ; Mengyuan LI ; Ruimin ZHU ; Juanjuan HU ; Rongfei WEI ; Kejuan LI ; Li JIANG ; Ran GAO
Chinese Journal of Comparative Medicine 2017;27(8):60-65
Objective To establish a patient-derived xenografts (PDX) mouse model of liver cancer (LC) and to explore its role in precision medicine.Methods PDX model was established by subcutaneous implantation of tumor tissues in NCG mice.The morphological structure of tumor tissue was exaimed using HE staining.Fifteen BALB/c nude mice were subcutaneously inoculated with tumor cell suspension from the PDX models.The xenograft mice were randomly divided into 5-fluorouracil (5-FU) group, sorafenib group and negative control group.The tumor volume and body weight of the tumor-bearing mice were measured regularly, the tumor inhibition rate was calculated and the curative effect was evaluated.Results The success rate was 33.3% (6/18) in the establishment of liver cancer PDX mouse model, and the model well retained the characteristics of the primary tumor.In one case of PDX mouse model, the tumor inhibition rates of 5-FU and sorafenib group were 63.7% and 29.6%, with a statistically significant differece between them (P< 0.05), and there was no significant difference between the sorafenib group and negative control group, consistent with clinical observation.Conclusions The PDX mouse model of liver cancer can maintain the histological structure of primary tumor, and can be applied to precision medicine for patients with liver cancer.
10.Comparison of hierarchical management core competence training of ICU nurses in China and England
Shouzhen CHENG ; Mengyuan WANG ; Yingxue GAO ; Mingrong GAO ; Ying ZHONG ; Jiahui. LUO
Chinese Journal of Practical Nursing 2018;34(17):1317-1321
ICU nursing is a vitalpartof the development of specialized nursing in China, and the standardized training of the corecompetence of ICU nurses is the primary goal of the development of ICU nursing. This paper took the First Affiliated Hospital of Sun Yat-sen University in China and the Royal Free Hospital in London, the United Kingdom as example, and compared the hierarchical management and core competencies training status of ICU nurses in China and England to find out the similarities and differences, and then to put forward valuable suggestions for hierarchical management and the core competence training of ICU nurses in China.