OBJECTIVE To standardize the main processes and related technical links of the clinical comprehensive evaluation of drugs， and provide guidance and reference for improving the quality of comprehensive evaluation evidence and its transformation and application value. METHODS The construction of Guideline for the Workflow of Clinical Comprehensive Evaluation of Drugs was based on the standard guideline formulation method of the World Health Organization （WHO）， strictly followed the latest definition of guidelines by the Institute of Medicine of the National Academy of Sciences of the United States， and conformed to the six major areas of the Guideline Research and Evaluation Tool Ⅱ. Delphi method was adopted to construct the research questions； research evidence was established by applying the research methods of evidence-based medicine. The evidence quality classification system of the Chinese Evidence-Based Medicine Center was adopted for evidence classification and evaluation. The recommendation strength was determined by the recommendation strength classification standard formulated by the Oxford University Evidence-Based Medicine Center， and the recommendation opinions were formed through the expert consensus method. RESULTS & CONCLUSIONS The Guideline for the Workflow of Clinical Comprehensive Evaluation of Drugs covers 4 major categories of research questions， including topic selection， evaluation implementation， evidence evaluation， and application and transformation of results. The formulation of this guideline has standardized the technical links of the entire process of clinical comprehensive evaluation of drugs， which can effectively guide the high-quality and high-efficient development of this work， enhance the standardized output and transformation application value of evaluation evidence， and provide high-quality evidence support for the scientific decision-making of health and the rationalization of clinical medication.

Objective To explore the role and mechanism of exosomes derived from bone marrow mesenchymal stem cells(BMSCs)overexpressing transglutaminase 2(TGM2)in lactate-mediated oxidative stress-induced apoptosis in nucleus pulposus cells.Methods Twenty 6-week-old male SD rats(350±50 g)were used to extract nucleus pulposus cells.Lactic acid was utilized to establish a model of oxidative stress-induced apoptosis in primary nucleus pulposus cells.Lentiviral vectors carrying TGM2 were employed to transfect BMSCs to overexpress the protein,and then exosomes were extracted from the BMSCs and identified.Subsequently,to investigate the potential mechanisms of TGM2-overexpressing exosomes against oxidative stress-induced apoptosis in nucleus pulposus cells,flow cytometry was used to detect the production of reactive oxygen species(ROS),and Western blotting and immunofluorescence assay were applied to measure the expression levels of Bax,Bcl-2,Cleaved-Caspase3,p-PI3K,p-Akt,n-Nrf2,c-Nrf2 and HO-1,and Nrf2 nuclear translocation.The rate of oxidative stress-induced apoptosis in nucleus pulposus cells and changes in the PI3K/Akt/Nrf2 pathway were assessed in 4 groups of cells(n=3):control group,lactate treatment group,lactate+control exosome group,and lactate+TGM2-overexpressing exosomes group.Results Lactic acid of 10 mmol/L was found to be more effective in inducing oxidative stress-induced apoptosis in nucleus pulposus cells(P＜0.05).Both control BMSCs and TGM2-overexpressing BMSCs could produce exosomes and stably overexpress TGM2(P＜0.05,P＜0.01).Compared with the lactate treatment group,the nucleus pulposus cells in the lactate+TGM2-overexpressing exosome group and the lactate+control exosome group showed reduced apoptotic rates(P＜0.05),as well as decreased ROS level(P＜0.05).The lactate+TGM2-overexpressing exosome group exhibited better anti-apoptosis and ROS accumulation effects(P＜0.05).Additionally,the expression of Bax and Cleaved-Caspase3 was decreased(P＜0.05),that of Bcl-2 protein was increased(P＜0.05),the phosphorylation of PI3K/Akt was enhanced(P＜0.05),and both Nrf2 nuclear translocation and the expression of the antioxidant stress factor HO-1 were increased in nucleus pulposus cells.The lactic acid+TGM2-overexpressing exosome group showed stronger anti-apoptotic effects and activation of the PI3K/Akt/Nrf2 pathway(P＜0.05).Conclusion Exosomes derived from BMSCs overexpressing TGM2 can inhibit lactate-mediated oxidative stress-induced apoptosis in nucleus pulposus cells,and the mechanism may be related to the activation of the PI3K/Akt/Nrf2 pathway.

Objective To investigate the effect and mechanism of Clara cell secretory protein (CC16 ) on lung injury of traumatic shock rats.Methods Thirty SD male rats were randomly assigned to three groups:sham-operation group,control group and CC16-treatment group.The rat model of traumatic shock was used in this study.Rats of sham-operation group were operated,but not treated by depletion and recovery.Rats of control group inhaled the physiological saline before fluid resuscitation.Rats of CC16-treatment group inhaled 0.1 μg/mL of recombinant human Clara cell secretory protein (rh-CCSP)before fluid resuscitation.Arterial blood gas analysis and wet/dry weight were detected in each group.The contents of malondialdehyde (MDA),myeloperoxidase (MPO)of lung tissues and lung tissue pathology changes were also studied. Results Compared with control group,pH and PaO2 value in CC16-treatment group increased significantly.Meanwhile,BE value and lung wet/dry weight ratio also decreased significantly (P<0.05).Compared with control group,contents of MDA and MPO in CC16-treatment group decreased significantly (P<0.05).Lung tissue pathology improved in the CC16 group compared with that of control group (P <0.05 ).Conclusion Inhaled Rh-CCSP may have potential protective effect on lung injury tissues of traumatic shock rats model through its antioxidative effect.

Objective To discuss the effective nursing method of severe erythema multiforma exudativum children patients complicated with bronchopneumonia.Methods Two cases of erythema multiforma exudativum were reviewed,and the nursing methods were summarized,including protective isolation,care of wound surface,care of intravenous infusions,psychological care,oral care,eyes care,perineal care,care of fever,and discharge instructions.Results Two children patients were both cured.Conclusions For severe erythema multiforma exudativum children patients complicated with bronchopneumonia,proper nursing method and careful observation can decrease the complications and help patients to cure quickly.

ObjectiveTo evaluate the role of Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway in attenuation of myocardial ischemia-reperfusion (I/R) injury by tetramethylpyrazine in rats.MethodsSixty-four healthy male Wistar rats weighing 250-300 g were randomly divided into 4 groups( n ＝ 16 each):sham operation group (group S),myocardial I/R group(group I/R),teramethylpyrazine group (group T) and AG,490( a JAK2 inhibitor) group (group AG).Myocardial I/R was induced by 30 min occlusion of left anterior desecending coronary artery (LAD) followed by 120 min reperfusion in groups I/R,T and A.In groups T and A teramethylpyrazine 20 mg/kg was injected iv 20 min before LAD occlusion.In group A AG490 3 tμg/g was injected iv at 5 min before reperfusion.Blood samples were then taken from inferior vena cava at 120 min of reperfusion for measurement of serum creatine phosphokinase (CK) and lactose dehydrogenase (LDH) activities.Myocardial infarct size was then measured and myocardial tissue was obtained for microscopic examination.ResultsSerum CK and LDH activities were significantly higher in group I/R than in group S.Pretreatment with tetramethylpyrazine significantly decreased myocardial infarct size and I/R-induced increase in serum CK and LDH activities and histologic damage.The protective effect of tetramethylpyrazine against myocardial 1/R injury was attenuated by postconditioning with AG490.ConclusionJAK2/STAT3 signaling pathway is involved in attenuation of myocardial I/R injury by tetramethyl pyrazine in rats.

Summary: The effect of rosiglitazone as the ligand of peroxisome proliferator-activated receptorγ (PPARγ) inhibiting the TLR4 expression in ischemic lobes in partial hepatic ischemia/reperfusion injury (IRI) in BABL/C mice and the action of rosiglitazone inhibiting the TLR4 receptor-mediated inherent immune response were investigated. The model of the mouse partial hepatic ischemia/reperfusion injury was established. All the animals were randomly divided into 3 groups: rosiglitazone group, vehicle (dimethylsulphoxide, DMSO) group and sham operation group. The hepatic samples were collected when mice were sacrificed 0, 2, 4 and 6 h after reperfusion following 1h ischemia to analyze the acute phase of hepatic IRJ. The dynamic expression of TIR4 mRNA was detected quantitatively by real-time-PCR, and the levels of TNF-α, IL-10 and ALT in portal vein were determined in all groups. After restoration of blood supply, the expression of TLR4 mRNA in ischemic lobes was detected in 0, 2, 4 and 6h after reperfusion following 1h ischemia in rosiglitazone group and vehicle group. The most intensive expression of TLR4 mRNA was present at 4 h after reperfusion in ischemic lobes in vehicle group. As compared with vehicle group, the expression of TLR4 mRNA in ischemic lobes in rosiglitazone group was significantly decreased at 4h after reperfusion. The level of IL-10 in portal vein was markedly up-regulated in rosiglitazone group as compared with vehicle group. Contrarily, the levels of TNF-α and ALT in portal vein were markedly down-regulated in rosiglitazone group as compared with vehicle group at every time point in mouse partial hepatic IRI model. Rosiglitazone could alleviate the hepatic IRI by inhibiting TLR4 receptor-mediated inherent immune response.