AIM: To explore whether free radicals participate in cerebral ischemic tolerance and the up-regula-tion of p38 MAPK and ERK signaling pathways in rats induced by limb ischemic preconditioning (LIP). METHODS: A total of 128 Wistar rats with permanent occlusion of bilateral vertebral arteries were randomly divided into sham group (n=16), cerebral ischemia (CI) group (n=16), LIP+CI group (n=16), DMTU (a free radical scavenger)+LIP+CI group (n=64) and DMTU+sham group (n=16). Six rats in each group were used to observe the delayed neuronal death (DND) in hippocampal CA1 region by thionin staining at 7 d after the end of operation. Other 10 rats in each group were used to de-tect the expression of p38 MAPK and ERK in hippocampal CA1 region by immunohistochemistry and Western blot. RE-SULTS: Lethal CI resulted in obvious DND in hippocampal CA1 region. However, LIP reversed the above injurious changes, represented by the decrease in histological grade and the increase in neuronal density compared with CI group (P<0. 01). Moreover, LIP significantly up-regulated the expression of p38 MAPK and ERK in hippocampal CA1 region com-pared with CI group (P<0. 01). Administration of free radical scavenger DMTU via femoral vein before LIP partially re-versed the neuroprotective effect of LIP, and blocked the up-regulation of p38 MAPK and ERK expression in hippocampal CA1 region in rats compared with LIP+CI group (P<0. 01). CONCLUSION: Free radicals are involved in the neuropro-tection and up-regulation of p38 MAPK and ERK expression induced by LIP in rats.

Macrolide and corticosteroid resistance has been reported in patients with Mycoplasma pneumoniae (MP) pneumonia (MPP). MP clearance is difficult to achieve through antibiotic treatment in sensitive patients with severe MPP (SMPP). SMPP in children might progress to airway remodeling and even bronchiolitis/bronchitis obliterans. Therefore, identifying serum biomarkers that indicate MPP progression and exploring new targeted drugs for SMPP treatment require urgency. In this study, serum samples were collected from patients with general MPP (GMPP) and SMPP to conduct proteomics profiling. The Fc fragment of the IgG-binding protein (FCGBP) was identified as the most promising indicator of SMPP. Biological enrichment analysis indicated uncontrolled inflammation in SMPP. ELISA results proved that the FCGBP level in patients with SMPP was substantially higher than that in patients with GMPP. Furthermore, the FCGBP levels showed a decreasing trend in patients with GMPP but the opposite trend in patients with SMPP during disease progression. Connectivity map analyses identified 25 possible targeted drugs for SMPP treatment. Among them, a mechanistic target of rapamycin kinase (mTOR) inhibitor, which is a macrolide compound and a cell proliferation inhibitor, was the most promising candidate for targeting SMPP. To our knowledge, this study was the first proteomics-based characterization of patients with SMPP and GMPP.
Biomarkers ; Carrier Proteins ; Child ; Humans ; Immunoglobulin Fc Fragments ; Immunoglobulin G ; Macrolides ; Mycoplasma pneumoniae ; Pneumonia, Mycoplasma/drug therapy* ; Proteomics

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

Objective To observe the effects and regulatory mechanism of rotavirus infection on the expression and bioactivity of Na+/H+exchanger 3 (NHE3) on Caco-2 cells. Methods A cell model of Caco-2 cells expressing NHE3 was constructed. Four groups were set up,which were control(CTL) group, rotavirus(RV) infection group, Cdc42 inhibitor (Pirl-1) group and Pirl-1+RV group. Bioactivity and ex-pression of NHE3 on the surface of Caco-2 cells were determined by BCECF-AM and biotinylation method, respectively. Expression of Cdc42 protein was measured by Western blot. Co-immunoprecipitation was per-formed to detect the interaction between NHE3 and Cdc42. Results Compared with the CTL group,RV in-fection significantly inhibited the bioactivity and expression of NHE3 on Caco-2 cells. These inhibitory effects were antagonized by Pirl-1. Moreover,RV infection enhanced the expression of Cdc42 protein and promoted the interaction between NHE3 and Cdc42, which were also antagonized by Pirl-1. Conclusion RV infec-tion might regulate the expression and bioactivity of NHE3 through Cdc42-dependent endocytosis pathway.

Objective To explore learning burnout psychology and professional identity of nursing students studying at the traditional Chinese medicine college.Methods 200 students were surveyed by learning burnout and professional commitment scale.Results Learning burnout level of nursing students was higher,depression scores were the highest in four dimensions.Professional commitment kept within medium level.The professional identity was low.The inheritance commitment score was the lowest in each dimension analysis.Conclusions Schools and teachers should take corresponding measures to improve professional commitment level,so as to reduce learning burnout level and ensure long-term stable development of nursing profession.

OBJECTIVETo evaluate the effect of infiltration resin on masking white spot lesions by assessing the change in the white spot area.

METHODSSeventy-four maxillary anterior teeth with post-orthodontic decalcification teeth were investigated in this study. All teeth were treated with infiltration resin according to manufacturer recommendation. Standardized digital images were taken before, immediately after, and one week after treatment. The results were classified into three groups: com-pletely masked, partially masked and unchanged. The images of partially masked teeth were analyzed using an image analysis software. The size of the white spot lesion (W) and the whole-tooth facial surface (T) were measured, and the W:Tratio (%) was calculated. Statistical evaluation of the lesions was performed using Kruskal-Wallis and Mann-Whitney tests.

RESULTSAmong the 74 teeth, 20 (27%) teeth were classified as completely masked and 54 (73%) teeth were classified as partially masked; no tooth was unchanged. The W:T ratio significantly decreased from 39.28% before treatment to 9.46% after treatment (P < 0.05).

CONCLUSIONResin infiltration is an effective treatment for masking white spot lesion. However, the masking effect depends on the lesion depth and activity.

Objective To evaluate the effect of infiltration resin on masking white spot lesions by assessing the change in the white spot area. Methods Seventy-four maxillary anterior teeth with post-orthodontic decalcification teeth were investi-gated in this study. All teeth were treated with infiltration resin according to manufacturer recommendation. Standardized digital images were taken before, immediately after, and one week after treatment. The results were classified into three groups: com-pletely masked, partially masked and unchanged. The images of partially masked teeth were analyzed using an image analysis software. The size of the white spot lesion (W) and the whole-tooth facial surface (T) were measured, and the W︰T ratio(%) was calculated. Statistical evaluation of the lesions was performed using Kruskal-Wallis and Mann-Whitney tests. Results Among the 74 teeth, 20 (27%) teeth were classified as completely masked and 54 (73%) teeth were classified as partially masked; no tooth was unchanged. The W︰T ratio significantly decreased from 39.28% before treatment to 9.46% after treat-ment (P<0.05). Conclusion Resin infiltration is an effective treatment for masking white spot lesion. However, the masking effect depends on the lesion depth and activity.

Objective To investigate the pattern of the cerebral white matter lesions in patients with mild and moderate Alzheimer's disease(AD)and healthy controls using proton magnetic resonance spectroscopy(~1H-MRS)and diffusion tensor imaging(DTI).Methods Twenty AD patients and Twenty healthy controls were recrnited.All subjects underwent clinical examination,neuropsychological assessment.The quantitative analysis of N-acetylaspartate(NAA),myoinositol(mI),Chotine(Cho)and Creatine(Cr)resonance signals in region of interests(ROIs)located in the paraventricular white matter region bilaterally were measured.Ratios of NAA/Cr,mI/Cr and Cho/Cr were calculated in two groups.In addition,conventional MRI and DTI scanning were received,fractional anisotropy(FA)and mean diffusivity(MD)values of white matter in the same regions were measured respectively.Results No significant difference between two groups were observed in NAA/Cr ratio(P＞0.05).A significantly increased mI/Cr and Cho/Cr were found in AD patients than in controls(P＜0.05).FA and MD values in AD patients were 0.470±0.082 and 0.771±0.099,and in controls were 0.539±0.068 and 0.691±0.064,respectively.FA value decreased significantly in AD patients(P＜0.05),M D value increased significantly in AD patients(P＜0.05).After controlling for age-related,partial correlation analysis revealed a negtive correlation between mI/Cr and FA value in the patients with AD(P＜0.05).No correlation between mI/Cr and MD was found(P＞0.05).Conclusion The results suggest that not only the gray matter is injured,but also the white matter is abnormal in AD patients.Combining ~1H-MRS with DTI alterations could provide the valuable informations about white matter lesions in AD patients.

BACKGROUND: A new method, i.e., heparin slow-release stent implantation combined with myocardium drilling, is discovered for myocardial revasculadzation, which remarkably improves myocardial perfusion. OBJECTIVE: To investigate the effect of heparin slow-release stent implantation combined with myocardium drilling on myocardial regeneration of pigs with acute myocardial infarction.METHODS: Anterior descending coronary of pig was ligated to induce myocardial infarction model, which was randomly divided into control and implantation groups, with 6 pigs for each group. Self-made borer was used in the implantation group to drill two transmural channels (3.5 mm diameter) on epicardium. A heparin slow-release stent was fixed in the transmural channel. Following intravenous injection, BrdU was used to label DNA duplication so as to observe stromal cell-derived factor-1 (SDF-1) mRNA expression, myocardial perfusion, newborn yocardium, and heart function prior to and following implantation. RESULTS AND CONCLUSION: As compared with control group, SDF-1 expression was enhanced in the implantation group at 6 weeks after stent implantation (P < 0.001), perfusion mass defect percentage was significantly decreased (P < 0.001), ejection fraction of left ventricle was increased (P < 0.05), newborn myocardium was increased (P< 0.001), and survival myocardium in the ischemic region was increased (P < 0.001). The results demonstrated that heparin slow-release stent implantation combined with myocardium drilling could repair damaged myocardial cells and improve heart function through increasing SDF-1 expression and myocardial perfusion.

AIM: To explore the role of NO/ inducible nitric oxide synthase (iNOS) in the metabotromi glutamate receptor 2/3C (mGluR2/3) mediated-brain ischemic tolerance induced by cerebral ischemic preconditioning (CIP), and to observe the influences of α-methyl- (4-tetrazolyl- phenyl) glycine (MTPG), an antagonist of mGluR2/3, on the expression of iNOS during the induction of brain ischemic tolerance. METHODS: Thirty-six Sprague-Dawley rats were subjected to four vessel occluding global brain ischemic model. Thionin staining and immunohistochemistry were used for neuropathological evaluation and assay of iNOS expression in the hippocampal CA1 subregion of the rats. RESULTS: In the sham group, weak expression of iNOS was detected. The expression of iNOS in the CIP and CIP+ischemic insult groups were increased significantly compared with that in the sham group. Administration of MTPG via lateral cerebral ventricle 20 min before CIP blocked the up-regulation of iNOS induced by CIP, but had no influence on the pyramidal neuron survival. However, in the MTPG+CIP+ischemic insult group, the expression of iNOS was extremely intensive compared to that in CIP and MTPG+CIP groups. Importantly, this up-regulation was accompanied with obvious delayed neuronal death. CONCLUSION: NO/iNOS pathway plays an important role in the process of mGluR2/3 mediated-brain ischemic tolerance induced by CIP.