AIM:To explore the expression of anoctamin 1 (ANO1), one of calcium-activated chloride chan-nels ( CaCCs) , in mouse cardiomyocytes and its functional properties.METHODS:The cardiomyocytes from the myocar-dial tissues of C57BL/6 mice were isolated with enzyme and purified by the differential adherent method.The cells were stained with monoclonal anti-sarcomeric actin and Cy3 to evaluate the purity of the myocardial cells.RT-PCR was used to detect the mRNA expression of ANO1 in the mouse cardiomyocytes.The protein expression of ANO1 in the mouse cardio-myocytes was determined by Western blotting analysis.The fluorescence quenching kinetics experiment was used to identify the ion transport properties of ANO1 in the mouse cardiomyocytes.RESULTS: The results of RT-PCR confirmed that ANO1 was expressed in freshly isolated myocardial cells.The results of Western blotting clearly demonstrated the protein expression of ANO1 in primarily cultured myocardial cells.Fluorescence quenching kinetics experiment on freshly isolated single myocardial cell revealed a pronounced outward rectifying property of the ANO1.The functional properties were simi-lar to the classic CaCCs.CONCLUSION:ANO1 expression was identified in the mouse myocardial cells.The function of CaCCs was generated by ANO1, suggesting that ANO1 is the molecular basis of CaCCs.

Objective To detect the location of histamine in peripheral sympathetic nerves of guinea pig. Methods Histidine decarboxylase mRNA was detected using in situ hybridization histochemistry with specific oligonucleotide probe,while histamine and tyrosine hydroxylase were detected using double labeled immunohistochemistry with anti-histamine antibody and anti-tyrosine hydroxylase antibody in the superior cervical ganglia of guinea pig. Results The histidine decarboxylase mRNA hybridization signal were detected in both of large and small cells.The TH immunoreactive substance distributed in cytoplasm steadly,but lacked in the nuclei,while the histamine immunoreactive substance distributed in cytoplasm nearby the plasmalemma.After chemical destroy of the guinea pig SCG′s neuron with 6-OHDA,the immunoreactive materials were hardly detected.Conclusion Because the histidine decarboxylase is the only enzyme which catalyzes histidine into histamine,histamine may be synthesized and coexisted with monoaminergic neurotransmitters in the superior cervical ganglia of guinea pig.

Objective To test experimentally Fluoro-Jade B(FJB) stain method for detecting degeneration of neurons in the basal ganglia. Methods Kainic acid(KA)-lesion model by stereotaxical injection of KA into striatum of rats,MPTP-lesion model by injection of MPTP into intraperitoneal cavity of mice,as well as KA-lesion model of cultured striatal cells were firstly prepared.FJB stain dye was then used to visualize degeneration of neurons in above KA-or MPTP-lesion models. Results KA-or MPTP-induced degenerative neurons including cell bodies and processes could be clearly visualized by FJB stain dye.In the brain sections,FJB-positive stained degenerative neurons were numerously observed in the striatum of KA-lesion rats and the substantia nigra pars compacta of MPTP-treated mice,but not detected in the control animals.Moreover,degenerative neurons were also detected with FJB stain in cultured striatal neurons.Semi-quantitative analysis on percentage(?s) of FJB-positive neurons constituting total cultured striatal neurons in unit area showed that degenerative neurons of KA-lesion group (8.42?1.09)% was evidently more than that of controls (3.42?0.45)%,P

Objective To observe the effect and safety of the endovascular hypothermia through hypothermic intravenous infusion device in the treatment of patients with severe craniocerebral injury compared with conventional surface cooling. Methods A total of 66 cases of patients with severe cranio-cerebral injury were randomly divided into the observation group and the control group with 33 cases in each group according to envelop randomization. The control group received surface cooling, and the observation group was given surface cooling plus endovascular hypothermia through hypothermic intravenous infusion device. The target temperature was 35 ℃maintained for 3-5 days, and natural rewarming was applied at the speed of 0.1-0.5℃/h to 36.0-37.3℃. The time to reach target temperature, the constant stability, the incidence rate and severity of complication such as shiver, arrhythmia, skin injury and agitation were recorded and compared between two groups, as well as the heart rate, breathing rate, pulse rate, blood pressure and Glasgow Coma Scale (GCS) scores after 72 h of treatment. Glasgow Outcome Scale scores 30 days after treatment and nursing workload were also calculated and compared. Results The cooling speed, time to reach target temperature and the ability to maintain at 35℃were (1.3±0.2)℃/h, (2.3±0.2) h and (6.5± 1.8)%in the observation group, respectively, compared with (0.5±0.1)℃/h, (3.6±0.6) h and (11.3±2.2)%in the control group, which had significant differences (t=1.862, 2.112, 2.408,P < 0.05). The occurrence rates of shiver, arrhythmia, skin damage and dysphoria and restlessness in the observation group were 33.33%(11/33), 9.09%(3/33), 6.06%(2/33) and 27.27%(9/33), respectively, which were much lower than those in the control group 84.85%(28/33), 15.15%(5/33), 33.33%(11/33), 54.55%(18/33),χ2=1.764-2.733,P<0.05. The heart rate, breathing rate, pulse rate, systolic blood pressure and GCS score after 72 h of treatment were(68.31 ± 3.73)times/min,(16.60 ± 1.52)times/min,(136.35 ± 3.71)mmHg(1 mmHg=0.133 kPa),(34.61 ± 1.05)℃, (9.91±4.05)points in the observation group, while(58.31±3.62)times/min,(19.81±1.83)times/min,(150.66± 2.70)mmHg,(35.65 ± 1.36)℃,(7.63 ± 3.17)points in the control group, and there were significant differences between two groups(t=2.275-3.035, P < 0.05).Besides, the ice-changing ice and turning-over time in the observation group were both remarkably reduced compared with control group, (14.03±3.11) min/h vs (38.12± 2.70) min/h (t=3.356, P<0.05) , (15.08±3.07) min/h vs (26.16±2.54) min/h ( t=3.021, P<0.05). Patients with good recovery, mild disability, severe disability, death in the observation group were 16, 13, 3 and 1 case, while 6, 11, 9, 7 cases in the control group (χ2=2.351,P < 0.05). Conclusions The endovascular hypothermia through hypothermic intravenous infusion device can rapidly reduce and effectively maintain target temperature, reduce the incidence rate of complication, improve the vital signs and decrease the nursing workload in order to improve neurological outcome in the treatment of patients with severe craniocerebral injury.

Objective:To explore the association between chest high resolution CT (HRCT) scoring and prognostic factors of interstitial lung disease (ILD) in patients with anti-melanoma differentiation-associated gene 5 (MDA5) antibody-positive dermatomyositis (DM).Methods:The patients with DM admitted to Kailuan General Hospital between January 2017 and December 2019 were included into the study including 13 patients with positiveanti-MDA5 antibody (7 survivors, 6 deaths) and 18 patients with anti-synthase (ARS)-antibody positive. All patients underwent chest HRCT prior to treatment. The consolidation, ground-glass opacity (GGO) and fibrosis were scored to assess HRCT findings. The clinical manifestations were compared between the two groups. Cox regression analysis adjusted for age and sex was used to determine the prognostic factors for anti-MDA5 antibody-related ILD.Results:Compared with ARS patients, glutamyl transferase (GGT) and ferritin levels were significantly higher in MDA5-ILD patients [70.0(37.0, 122.5) vs 21.0(16.5, 33.5), Z=-3.37, P=0.001; 977.0(502.5, 1 366.0) vs 307.1(72.3, 546.9) , Z=-3.44, P=0.001]. The cumulative survival rate was significantly lower in patients with positive anti-MDA5 antibody than in those with positive anti-ARS antibody (100% vs 70%, P=0.001). The DM complicated with acute/subacute interstitial pneumonia (A/SIP) were found to significantly relate to death. There were no significant differences in chest HRCT scoringbetween the survivors and the deceased patients [ HR=1.08, 95% CI(0.95, 1.23), P=0.229; HR=0.97, 95% CI(0.72, 1.30), P=0.814]. Conclusion:Anti-MDA5 antibody is an important index for early diagnosis of DM complicated with acute/subacute interstitial pneumonia (A/SIP). The chest HRCT scoreis is not associated with the prognosis of anti-MDA5 antibody-related ILD patients.

Agmatine is an endogenous amine synthesized from the decarboxylation of arginine.It has a rich biological effects and presents in plants, bacteria and mammalian tissues.Agmatine is highly polar and has a low molecular weight.There are no UV and fluorescent absorption groups in agmatine structure, so it is difficult to gasify.In addition, the content of endogenous agmatine is low, and there are many interference components in biological samples, and the general method is difficult to detect.The pre-column derivatization of agmatine with fluorescence reagents not only increase the molecular weight of agmatine, but also make them with fluorescence, which greatly improve the detection sensitivity and enable the endogenous agmatine to be well separated.It is an important method to determine the content of agmatine.In this paper, several kinds of precolumn derivatives are summarized, and the advantages and disadvantages of various derivatives, the best derivation conditions and detection methods were also analyzed comprehensively.The aim is to determine the content of agmatine and to provide methods and ideas for its biological research.

Objective To research sorption of local anesthetic by fat emulsion with different content of lipid in vitro and then to discuss the mechanism of action of fat emulsion for treatment of local anesthetic toxicity. Methods 10%,20%,30% fat emulsion was added to the bupivacaine hydrochloride and ropivacaine hydrochloride.After the mixture was vortexed for 10 mins,it was vibrated in thermostatic water bath at 37 ℃ for 15 h,then centrifugated and got the aqueous phase to HPLC test.The chromatography was carried on a C18column(4.6 mm×250 mm,5 μm ),the mobile phase was composed of acetonitrile-natrium biphosphoricum(25:75),the flow rate was 1 mL·min-1,the column temperature was 30 ℃ and the detection wavelength was set at 210 nm. Results Under the condition of this test,the linear ranges and linear equations for bupivacaine hydrochloride and ropivacaine hydrochloride were 0.3-3.0 μg·mL-1,Y=0.093 6X-0.017 3(r=0.999 6),and 0.3-3.0 μg·mL-1,Y=0.086 6X-0.022 3(r=0.999 1) respectively.The lowest limit of quantitation were both 0.05 μg·mL-1(S/N>3),and the ingredients showed good relationships between the peak area and the concentration.Sample reproducibility was good because the test sample was stable in 5 h.The extraction rates of bupivacaine hydrochloride by 10%,20%,30% fat emulsion were 46.0%,70. 4%,89.2% respectively.The extraction rates of ropivacaine hydrochloride by 10%,20%,30% fat emulsion were 51.3%,71.6%, 90.7% respectively. Conclusion Electric potential and the pH value of the 10%,20%,30% fat emulsion are similar,while the particle size increase slightly.Free bupivacaine hydrochloride and ropivacaine hydrochloride can be absorbed and with the increase of fat content,the extraction of local anesthetic is increased.The mechanism of action of fat for the treatment of local anesthetic toxicity maybe related to that fat emulsions can provided lipid,so that the excessive local anesthetic or those which had already distributesd in tissue can re-dispersed in lipid,and then the plasma concentration of local anesthetic is reduced,the toxicity is also reduced.

Objective: To develop an assessment questionnaire for screen based behaviors among primary and secondary school students, so as to provide a basis for comprehensive assessment among the population. Methods: Nineteen experts were invited to take part in this study from August to September 2022. The initial framework and item pool of the assessment questionnaire were developed based on literature review and a series of guidelines and standards issued by the national education and health authorities, as well as relevant domestic and foreign guidelines, and were combined with existing assessment tools that were used to evaluate screen based behavior among these age groups. Experts in related fields were selected for two rounds of Delphi consultation to determine the dimensions, items and corresponding weights of the assessment questionnaire. Results: The response rates from two rounds of consultation were 95.0% and 100.0% respectively, the opinion submission rates were 89.5% and 63.2% respectively, and the authority coefficient ( Cr ) was 0.87. An assessment questionnaire was finally designed, consisting in 44 items and four dimensions, namely screen use type and time, screen use related health behaviors, safety and civilization in screen use, and reasonable arrangement of screen and non screen activities. The coefficient of variation( CV ) of each item ranged from 0.00 to 0.19, and the Kendall s W increased from 0.22 to 0.34( P <0.01). Expert opinions were in agreement and credible. Conclusion The designed assessment questionnaire can be used to comprehensively evaluate screen based behavior among primary and middle school students, and provides a basis for subsequent intervention measures to improve screen based behavior among this population.

Objective To investigate the relationship between serum uric acid ( UA) level and brachial?ankle pulse wave velocity ( baPWV) in patients with systemic lupus erythematosus ( SLE) and lupus nephritis (LN)??Methods A total of 110 hospitalized,out?patient and healthy examinees from January 2017 to September 2017 were selected from Kailuan General Hospital??They were divided into three groups:(1)Fifty?five healthy controls were examined at the same time,and those who had no history of hypertension, myocardial infarction and stroke were excluded by physical examination??(2)Thirty?four SLE patients without LN were diagnosed according to the SLE classification standard revised by the American Society of Rheumatology ( ACR) in 1997,excluding those with lupus nephritis??( 3) 21 SLE patients with LN were diagnosed according to the SLE classification standard revised by the American Society of Rheumatology (ACR) in 1997??Pearson correlation coefficient and multivariate linear regression model were used to analyze the related factors affecting baPWV??Results The level of baPWV and the proportion of baPWV (≥1400 cm／s) in SLE without LN group and SLE with LN group were higher than those in healthy control group (all P＜0??05)??In SLE without LN group, baPWV was positively correlated with age, systolic blood pressure (SBP) and total cholesterol ( CHOL) ( r＝ 0??623,0??528,0??402, P＜0??01 or P＜0??05), and negatively correlated with blood uric acid(UA) ( r＝－0??371,P＜0??05),but the correlation was not significant??The correlation between UA and baPWV disappeared after after correction of age,SBP,diastolic blood pressure (DBP) by partial correlation analysis??In SLE with LN group,baPWV was positively correlated with SBP, DBP and serum creatinine ( Cr) ( r＝0??815, 0??725, 0??464, P＜0??01 or P＜0??05)??Multivariate stepwise regression analysis showed that SBP was independently correlated with baPWV in SLE group ( t＝2??54,P＝0??026); UA in SLE group without LN was independently negatively correlated with baPWV(t＝－2??96,P＝0??042); UA(t＝4??24,P＝0??013) and SBP(t＝7??70,P＝0??002) were independently positively correlated with baPWV in SLE group with LN??Logistic regression analysis showed that SLE was a risk factor for baPWV (≥1 400 cm／s),and the OR (95% CI) was 4??31 ( 1??56－11??88),P＝0??005,and there was statistical significance after adjusting for age,SBP,DBP,body mass index ( BMI)??However,UA was not a risk factor for baPWV (≥1 400 cm／s) (P values were 0??163 and 0??519,respectively)??Conclusion The degree of arteriosclerosis in SLE patients is higher than that in normal subjects,and the level of UA in SLE patients may be related to baPWV??

The construction of healthy schools is one of the key measures to implement the Outline of the Healthy China 2030 Plan. The establishment and implementation of the Guidelines for Healthy Primary and Secondary School Evaluation （Guidelines） is helpful for the large-scale promotion and standardization of healthy school construction by government departments in Shanghai and other major cities in China， thereby more students benefited from the Healthy China Initiative. The Guidelines integrate the efforts of multiple departments related to school health， based on internationally recognized content and strategies for health-promoting schools. They aim to foster students’ health development by combining the basic requirements for school health with priority projects for health promotion. Healthy school evaluation indicators and standardized requirements suitable for the current socio-economic development level of Shanghai have been developed， aligning with international concepts of health-promoting schools. They also reflect a high starting point and high level of investment and development in school health in Shanghai over the past decade. The Guidelines provide recommendations on the basic requirements， organization guarantees and management， social and culture environment， health promotion education， health services， and connection between school and communities， and families， of healthy school evaluation for primary and secondary schools. This paper aims to interpret the compilation process and main content of the Guidelines.