Objective To evaluate the effects of sevoflurane preconditioning on zonula occludens-1 (ZO-1) during myocardial ischemia-reperfusion (I/R) in rats in vitro.Methods Adult male Wistar rats,weighing 250-300 g,were anesthetized with intraperitoneal pentobarbital 30 mg/kg and heparinized.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95％ O2-5％ CO2 at 37 ℃.Eighteen isolated rat hearts were randomly assigned into 3 groups (n =6 each) using a random number table:control group (group C),group I/R and sevoflurane preconditioning group (group S).At 10 min of equilibration,the hearts were perfused with K-H solution for 110 min in group C,the hearts were perfused with K-H solution for 20 min,and then subjected to 30 min of ischemia followed by 60 min of reperfusion in group I/R,and the hearts were perfused with K-H solution saturated with 3％ sevoflurane for 15 min followed by 5 min washout,and then subjected to 30 min of ischemia followed by 60 min of reperfusion in group S.At the end of equilibration,immediately before ischemia,and at 15 and 60 min of reperfusion (T1,2),HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP),+ dp/dtmax and-dp/dtmax were recorded.The development of arrhythmias was recorded during reperfusion and scored.At 60 min of reperfusion,myocardial specimens were obtained from the apex of heart for determination of the expression of ZO-1 in myocardial tissues (by Western blot) and for observation of distribution of ZO-1 and connexin43 (Cx43) (by immunofluorescence).Results Compared with group C,HR,LVDP,+ dp/dtmax and-dp/dtmax were significantly decreased and LVEDP was increased at 15 and 60 min of reperfusion,scores of arrhythmia was increased,and ZO-1 expression was down-regulated in I/R group.Compared with group I/R,HR,LVDP,+ dp/dtmax and-dp/dtmax were significantly increased and LVEDP was decreased at 15 and 60 min of reperfusion,arrhythmia was decreased,and ZO-1 expression was up-regulated in group S.ZO-1 and Cx43 were co-localized at the intercalated disk.ZO-1 was redistributed in the lateralization of the membrane and co-localized with Cx43 in group I/R.The incidence of ZO-1 lateralization was significantly decreased in group S.Conclusion The mechanism by which sevoflurane preconditioning decreases reperfusion arrhythmia is related to inhibition of down-regulation of expression and redistribution of ZO-1 and inhibition of redistribution of Cx43 in rats.

Objective To evaluate the effects of sevoflurane preconditioning on wnt/glycogen synthase kinase-3 beta (GSK-3β)/β-catenin signaling pathway during myocardial ischemia-reperfusion (I/R) injury in rats in vitro.Methods Ault male Wistar rats,weighing 220-280 g,were heparinized and anesthetized with intraperitoneal 3％ pentobarbital 30 mg/kg.Their hearts were rapidly excised and perfused in a langendorff apparatus with oxygenated (95％ O2-5％ CO2) K-H solution at 37 ℃.After 15 min of equilibration,36 isolated hearts were randomly divided into 3 groups (n=12 each) using a random number table:sham operation group (group S),group I/R and sevoflurane preconditioning group (group SP).After 30 min of equilibration,the hearts were continuously perfused for 150 min in group S.The isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion.In SP group,the hearts were perfused for 15 min with K-H solution containing 2.4％ sevoflurane,followed by 5 min washout before reperfusion.At the end of equilibration and 30 min of reperfusion,HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and ± dp/dtmax were recorded.The severity of arrhythmias was assessed during reperfusion.At 60 min of reperfusion,3 hearts in each group were chosen for measurement of expression of wnt3a,phosphor-GSK-3β (p-GSK-3β) and β-catenin (by Western blot).At 120 min of reperfusion,6 hearts in each group were chosen for determination of myocardial infarct size by TTC staining.Results Compared with group S,HR,LVDP,+dp/dtmax and -dp/dtmax were significantly decreased,and LVEDP was increased at 30 min of reperfusion,arrhythmia scores and the percentage of myocardial infarct size were increased,and the expression of wnt3a,p-GSK-3β and β-catenin was down-regulated in I/R group.Compared with group I/R,HR,LVDP,+dp/dtmax and-dp/dtmax were significantly increased,and LVEDP was decreased at 30 min of reperfusion,arrhythmia scores and the percentage of myocardial infarct size were decreased,and the expression of wnt3a,p-GSK-3β and β-catenin was up-regulated in group SP.Conclusion Sevoflurane preconditioning attenuates myocardial I/R injury by activating wnt/GSK-3β/β-catenin signaling pathway in isolated rat hearts.

Objective:To investigate the effects and underlying mechanisms of XRCC3 on esophageal squamous-cell carcinoma (ESCC) radiotherapy response. Methods:Expression levels of XRCC3 were detected by reverse transcription PCR, Western blot, and immunohistochemistry. We knocked down XRCC3 with lentiviral infection in ESCC cells. Cell apoptosis was examined by flow cytom-etry. DNA damage and telomere dysfunction-induced foci were determined by immunofluorescence. Results:The expression levels of XRCC3 in ESCC cells and tissues were higher than those in normal esophageal epithelial cells and corresponding adjacent noncancer-ous esophageal tissues. Knockdown of XRCC3 in ESCC cells substantially increased the therapeutic efficacy of radiation. We demon-strated that the radiation resistance of XRCC3 was attributed to the XRCC3-maintaining telomere stability, which reduced ESCC cell death through radiation-induced apoptosis. Conclusion: Our data suggested that XRCC3 protects ESCC cells from ionizing radia-tion-induced DNA damage and death by enhancing telomere stability. Thus, XRCC3 can be used as a promising therapeutic target for ESCCs.

Objective To investigate the impacts of pyruvate kinase M2 isoform (PKM2) silencing on the radiosensitivity of lung adenocarcinoma cell line (A549 cells) and the radiation synergy of xenografts, and to explore their mechanisms. Methods Plasmid pshRNA?PKM2 for interference with PKM2 expression was transfected into A549 cells, and empty vector?transfected cells and untransfected cells were set as con?trols. The silencing efficiency of pshRNA?PKM2 and the expression level of microtubule?associated protein 1 light chain 3(LC3) were measured by Western blot assay. The radiosensitizing effects in A549 cells and xen?ografts after PKM2 silencing were determined by colony?forming assay and xenografts growth curves. Autoph?agy formation in A549 cells and xenografts was analyzed by transmission electron microscopy, and the ex?pression level of PKM2 in xenografts was measured by immunohistochemistry. Comparison between groups was made by Student′s t?test, and the body weights of nude mice and xenograft volumes were subjected to a?nalysis of variance for continuous variables. Results Stable A549 cell lines transfected with pshRNA?PKM2 were successfully produced. Transfection with pshRNA?PKM2 significantly down?regulated PKM2 expression in A549 cells and xenografts (P= 0?? 001;P= 0?? 000). The sensitizer enhancement ratios for A549 cells and xenografts were 1?? 47 and 2?? 00, respectively. Interference with PKM2 expression enhanced radiation?in duced autophagy formation and significantly increased the ratio of LC 3 ? II / I ( P= 0.000 1 ) . Conclusions Silencing of PKM2 expression may enhance the radiosensitivity of A549 cells and xenografts by regulation of autophagy, which holds promise for becoming an effective radiosensitizing target for non?small cell lung canc?er, but still needs to be confirmed by further studies.

A derivatization method for the determination of oligosaccharides with high sensitivity by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry(MALDI-TOF-MS) based on a labeling regent 1-(4-Cyanophenyl)-4-piperidinecarbohydrazide(CPH) was proposed. The hydrazones were generated by the reaction of the reducing ends of oligosaccharides and the hydrazide of CPH. The derivatives were directly analyzed by MALDI-TOF-MS. Reaction conditions such as reaction temperature, concentration of acetic acid and excess ratio of CPH regent were optimized. When the oligosaccharides reacted with 100 time excess of CPH at 100 ℃ and in 0.125% acetic acid (V/V), the yield was highest. The results showed the derivatization with CPH increased the sensitivity of MS detection and this method was suitable for the analysis of oligosaccharides′ compositions.

Objective: To study the clinical effect of high pressure oxygen and Butylphthalide in the recovery of cerebral metabolism after carbon monoxide poisoning. Methods: 84 patients treated from May 2014 to May 2016 in our hospital were selected. The subjects were randomly and equally divided into two groups. The control group adopted the conventional therapy and high pressure oxygen; on the basis, the observation group also took Butylphthalide. The clinical effect, duration of coma, recovery of consciousness, incidence rate of delayed encephalopathy was observed. After 1m of treatment, the HDS point was evaluated. Results: The total effective rate of control group (76.19%, 32/42) was lower than that of observation group (95.24%, 40/42) (P<0.05) . The duration of coma for observation group was shorter than that of control group. The percentage for patients with recovery of consciousness and incidence rate of delayed encephalopathy for observation group was better than that of control group (P<0.05) . The HDS point for observation group was even higher than that of control group (P<0.05) . Conclusion The high pressure oxygen and butylphthalide can improve the clinical effective rate, shorten the duration of coma and promote the patient’s recovery of consciousness. It is worthy of clinical promotion.

Objective To implement and evaluate evidence-based guidelines for enteral nutrition support in acute stroke patients with dysphagia.Methods This study is a prospective before and after comparison study.Collected 200 acute stroke patients with dysphagia and divided them into test group (trained medical staffs) and control group(untrained medical staffs) equally according to the time order.Two groups of 100 patients were surveyed using a checklist before and after implementation of 10 guidelines about nutrition support.Before the implementation of guidelines,the staffs were enforced training,and summarized regularly.Compliances with guidelines by doctors and nurses were compared,and outcomes of patients were assessed.Results Compared with the control group,the correct implementation of the project significantly improved in the experimental group on nutritional risk screening (92.0％,64.0％; x2 =22.840),nutritional supplements selection (80.0％,48.0％; x2 =22.220),nutrition infusion methods (90％,18％ ; x2 =1.040) and nutrition infusion adjustment (abdominal distension/adjusted:21/10,6/4;x2 =9.634,constipation/adjusted:41/40,57/53 ; x2 =5.122,all P ＜ 0.05).The mortality rate,poor prognosis and length of stay in department of neurology intensive care unit and in hospital were not significant different between the experimental group and the control group.The incidence of hospital-acquired pneumonia was significantly lower in the experimental group (44.3％) than that in the control group (67.5％,x2 =7.281,P =0.007),but other patient outcomes were unaffected significantly.Conclusion Implementation of evidence-based guidelines for enteral nutrition support in acute stroke patients with dysphagia is associated with improvements in clinical quality and selected patient outcomes.

Objective To investigate the colonization of Gram-negative bacilli carrying mcr-1 gene in intestinal tracts of inpatients and people having physical examination for further elucidating the molecular and epidemiological features of mcr-1 gene. Methods A total of 1263 and 750 fecal specimens were col-lected from inpatients in the First Affiliated Hospital of Guangzhou Medical University and people having physical examination in the Kingmed Physical Examination Centre, respectively. Drug-resistant bacteria were isolated using Maconkey agar supplemented with colistin. PCR was performed to detect the bacteria carrying mcr-1 gene. Multilocus sequence typing ( MLST) and enterobacterial repetitive intergenic consensus-PCR ( ERIC-PCR) were used for homology analysis. The transferability of mcr-1 gene was verified by plasmid transfer assays. Plasmids of mcr-1-carrying strains were typed by PCR-based replicon typing techniques. Twelve virulence-related genes were also detected by PCR. Results Ninety-two colistin-resistant strains were isolated from the 1263 samples from inpatients(7. 3％, 92/1263) and two of them were positive for mcr-1 gene ( one strain also carried the blaNDM-5 gene) . Thirty-six colistin-resistant strains were isolated from the 750 samples of physical examination group (4. 8％, 36/750) and one of them carried the mcr-1 gene. MLST analysis showed that three mcr-1-carrying Escherichia coli strains ( minimum inhibitory concentration of colistin:8 μg/ml) belonged to three different sequence types. Moreover, they exhibited different banding patterns in ERIC-PCR analysis. All of the mcr-1-carrying isolates could transfer mcr-1 gene to the recipient strains successfully. Six types of incompatibility plasmids were detected in the mcr-1-carrying isolates ( IncFⅡ, IncX2, IncHI2, IncFIB, IncX4 and IncX1). Virulence-related genes fimH, iutA and fyuA were detec-ted in all mcr-1-carrying Escherichia coli strains. Conclusions Colistin-resistant strains and mcr-1 gene are prevalent in inpatients and people having physical examination, which brings potential risk for the control of clinical infections.

Objective:To explore and analyze the value of CT texture combined with baseline data in lymph node metastasis of cN0 papillary thyroid microcarcinoma (PTMC), in order to provide evidence for the evaluation of lymph node metastasis of PTMC.Methods:106 patients with single stage cN0 papillary PTMC confirmed by operation and pathology in Department of Radiology and Hernia Department of Huzhou First People’s Hospital from Aug. 2020 to Aug. 2022 were selected for retrospective analysis. Metastatic lymph nodes and non-metastatic lymph nodes were delineated in CT images, and they were divided into a lymph node metastasis group (22 cases) and a group without lymph node metastasis (84 cases). At the same time, lymph node CT texture feature images were obtained in both groups. Logistic regression was used to analyze the relationship between CT texture and lymph node metastasis, and ROC curve was drawn to evaluate the predictive value of CT texture for lymph node metastasis in cN0 stage PTMC patients. The patients were divided into<1/4 group (51 cases), 1/4-<1/2 group (36 cases), and ≥1/2 group (19 cases) according to the different contact range between tumor and thyroid margin in CT signs. The baseline data and lymph metastasis distribution of each group were analyzed.Results:The results of single factor analysis showed that there were no differences in tumor location, tumor calcification or thyroglobulin in CT texture between LNM group and non-LNM group ( P>0.05). The incidence of tumor contact with thyroid margin, tumor diameter ≥7 mm, age<45 and male ratio of sex in LNM group were higher than those in non-LNM group ( P<0.05). Multivariate Logistic regression analysis showed that tumor was in contact with thyroid margin ( OR=6.080, 95% CI: 1.738-21.273), tumor diameter ≥7 mm ( OR=2.779, 95% CI: 1.135-6.805), age<45 years old ( OR=6.074, 95%CI: 1.980-18.636) and gender male ( OR=12.642, 95% CI: 3.125-51.136) were independent risk factors for lymph node metastasis in cN0 stage PTMC patients ( P<0.05). There were no significant differences in gender, age, tumor diameter, tumor location, tumor calcification or thyroglobulin among<1/4 group, 1/4-1/2 group and ≥1/2 group ( P>0.05). The incidence of LNM in ≥1/2 group was higher than that in<1/4 group and 1/4-1/2 group, and pairwise comparison between groups had statistical significance ( P<0.05) . Conclusions:It is an independent risk factor for lymph node metastasis in cN0 stage PTMC patients with tumor and thyroid boundary contact, tumor diameter ≥7 mm, age<45 years old, male, and the wider the tumor and thyroid boundary contact, the higher the risk of lymph node metastasis. CT texture analysis combined with baseline data has important reference value in predicting lymph node metastasis of cN0 TMPC.

Objective:To investigate the value of magnetization transfer imaging (MTI) and fat suppression T 2WI (FS-T 2WI) in predicting the clinical activity of Graves ophthalmopathy (GO). Methods:From October 2020 to July 2021, 64 GO patients were prospectively enrolled in the First Affiliated Hospital of Nanjing Medical University. According to the clinical activity score (CAS), the patients were divided into active group (CAS≥3, 39 patients and 78 eyes) and inactive group (CAS<3, 25 patients and 50 eyes). The coronal MTI and FS-T 2WI were scanned for pre-treatment assessment. Magnetization transfer ratio (MTR) of extraocular muscles, and signal intensity ratio (SIR) between extraocular muscles and temporalis were measured, respectively. The independent-sample t-test was used to compare the MTR and SIR between two groups. The correlations between MRI parameters and CAS were analyzed using Spearman correlation analysis. The receiver operating characteristic (ROC) curve was performed to evaluate the value of each and combined parameters for predicting the clinical activity of GO. The DeLong test was used to compare the area under the curve (AUC). Results:The MTR of active group and inactive group were 0.45±0.04 and 0.51±0.04, respectively, the difference was statistically significant ( t=7.62, P<0.001). The SIR were 3.4±0.6 and 2.6±0.5, respectively, and the difference was also statistically significant ( t=-8.20, P<0.001). MTR was negatively correlated with CAS ( r=-0.46, P<0.001), while SIR was positively correlated with CAS ( r=0.63, P<0.001). The AUC of MTR, SIR and the combination of MTR and SIR for predicting the clinical activity of GO were 0.840, 0.845 and 0.905, respectively. The combination of MTR and SIR showed higher performance than MTR or SIR alone, and the differences were statistically significant ( Z=2.61, P=0.009; Z=2.15, P=0.032). Conclusions:The quantitative parameters of MTI and FS-T 2WI, namely MTR and SIR, can be used to evaluate the clinical activity of GO. Integrating MTI and FS-T 2WI can improve the diagnostic efficiency.