Objective To investigate whether and what staining techniques are applied to the ultrathin sheet plastination slice and whether the stained specimen is of autofluorescences .Methods A cadaveric hand block was plastinated and then sectioned as a series of 300-400μm thick transverse sections .A total of 56 slices in total .Alternative sections were stained with hematoxylin -eosin staining ( HE) , Verhoeff -Van Gieson staining ( VVG) or methylene blue and azureⅡstaining(MA).The stained slices were examined under a light microscope and a confocal microscope .Results The plastinated slices were stained with the three staining methods .HE staining revealed the muscle and connective tissues were red or violet , bone was violet or blue;VVG staining showed the elastic fibers was black , the collagen was red , and other tissues were yellow .MA staining showed the tendon was violet , the bone was pink , cartilage was violet , and other tissues were purple.However, the intracellular structures appeared not very well stained .The collagen, elastin and muscular structures in the stained slices were observed under a confocal microscope .Conclusion The commonly used histology staining methods can be used to stain the ultrathin sheet plastination slices .The staining provides a better observation of various tissues in the slice than the unstained slice .After staining, those autofluorescent structures in the plastinated slice are detectable under a confocal microscope .

Objective To observe the protective effect of Zhilong Huoxue Tongyu Capsule(Zhilong Capsule)on myocardial ischemia reperfusion injury(MIRI)in mice,and explore its regulatory mechanism using metabolomics.Methods Using a random number table method,30 C57BL/6J mice were randomly divided into the following three groups:sham operation group,model group,and Zhilong Capsule group(6.24 g/kg),with 10 mice in each group.In mice in the model group and the Zhilong Capsule group,a mouse MIRI model was established by ligating the left anterior descending branch,while mice in the sham operation group underwent threading without ligation.The Zhilong Capsule group began modeling one week after pre-administration and continued to receive intragastric administration for two weeks after modeling once daily.The cardiac function,including the left ventricular ejection fraction(LVEF)and left ventricular fraction shortening(LVFS),was assessed by color echocardiography.The myocardial fibrosis and apoptosis were observed by Masson staining and TUNEL staining,respectively.Enzyme-linked immunosorbent assay was used to measure the serum contents of lactate dehydrogenase(LDH)and brain natriuretic peptide(BNP).Liquid chromatography-mass spectrometry combined with multivariate statistical method was performed for serum metabolite detection and identification analysis.Results Compared with the model group,the mice in the Zhilong Capsule group exhibited an increase in LVEF and LVFS,a reduction in cardiac tissue structure disorder,a decrease in myocardial fibrosis,a decrease in cell apoptosis rate,and a decrease in serum LDH and BNP contents(P<0.05).Metabolomics result showed that intervention with Zhilong Capsule significantly regulated 30 differential metabolites related to MIRI.Important metabolic pathways involved 20 pathways related to tyrosine metabolism,arginine and proline metabolism,and vitamin digestion and absorption.Conclusion Zhilong Capsule has a protective effect on MIRI,and it may achieve this effect by regulating pathways related to tyrosine metabolism,arginine and proline metabolism,and vitamin digestion and absorption.

Objective:To investigate the influencing factors of the survival in infants with congenital diaphragmatic hernia (CDH) and to develop a prediction model for CDH.Methods:Clinical data of 252 infants with a prenatal diagnosis of CDH in Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from January 2010 to December 2018 were retrospectively analyzed. Adverse outcomes were defined as neonatal death on discharge. Chi-square and t test were used for univariate analysis of 16 perinatal and five postnatal risk factors. Regression analysis was used to determine the independent predictors of survival. The receiver operating characteristics (ROC) curves of the risk factors for predicting the survival of CDH were drawn. A prediction model based on the combination of risk factors for predicting adverse outcomes of CDH was established using the cut-off value. ROC curves of the model were drawn and the area under curve (AUC), sensitivity, and specificity were calculated. Results:Out of the 252 patients, 173(68.7%) survived on discharge. Univariate analysis showed that lung-to-head ratio (LHR), polyhydramnios, right diaphragmatic hernia, liver herniation, intrathoracic stomach, premature birth, low birth weight, 5 min Apgar score <7, and amniotic fluid index (AFI) were significantly associated with the survival of CDH. Regression analysis showed that polyhydramnios ( OR=11.19,95% CI:2.83-45.33), liver herniation ( OR=2.81,95% CI:1.32-11.92), intrathoracic stomach ( OR=5.02, 95% CI:1.29-17.13), low birth weight ( OR=8.58,95% CI:1.59-45.01) and AFI ( OR=3.68, 95% CI:1.37-14.72) were independent risk factors for survival at discharge in children with CDH, while LHR ( OR=0.36, 95% CI:0.01-0.69) were protective factors. The predictive cut-off values of LHR, polyhydramnios, liver herniation, intrathoracic stomach, low birth weight, and AFI were 1.6, 1.0, 1.0, 1.0, 1.0, and 232.5 mm, respectively. The model based on the combination of the above indicators for predicting CDH adverse outcomes was shown with an AUC value of 0.904, predictive sensitivity of 0.747, and specificity of 0.896. Conclusions:In this study, LHR, liver herniation, intrathoracic stomach, polyhydramnios, low birth weight, and AFI are independent risk factors for CDH survival. The combination of prenatal and postnatal indicators is noted for a higher accuracy for predicting CDH survival.

OBJECTIVE To explore the effect of iron sulfur cluster binding protein 1(ISCA1)mag-netic field on calcium inflow.METHODS ① A plasmid containing the ISCA1、Magneto 2.0 gene sequence,and an empty plasmid was prepared for lentivirus packaging,with all the three plasmids carrying the mCherry gene.The infection efficiency of lentiviruses was assessed using fluorescence microscopy.② The immunoprecipitation technique was employed to ascertain the interactions between ISCA1 and cryptochrome(CRY1)as well as CRY2 proteins.③ The technique of live cell calcium imaging was used to detect the intracellular calcium influx in cells exhibiting high levels of ISCA1 or Magneto 2.0 expression under the influence of a magnetic field(40 mT,0.1Hz,90%duty cycle).RESULTS ① Red fluorescence was observed by fluorescence microscopy,indicating successful transfection of lentivirus.② The exogenous ISCA1 proteins exhibited no affinity towards the endogenous CRY1 or CRY2 proteins.③ The green fluorescence intensity of the Magneto 2.0 group increased by(1.8±0.5)times compared to the pre-magnetic addition level,indicating a significant influx of calcium ions.However,there was no significant difference in the green fluorescence intensity between the ISCA1 group and control group before and after magnetic addition.CONCLUSION Cells with exogenously high expres-sion of ISCA1 do not elicit calcium influx and exhibit no discernible magnetic induction following stimula-tion by this magnetic field.

Objective: To establish the norm of the Physical Activity afterschool Questionnaire for Preschooler(P-PAQ) in urban areas of China, so as to provide a basis for graded guidance from the family perspective and to improve children s physical activity levels. Methods: From October 2020 to January 2021, 6 267 children aged 3-6 years old were recruited from 40 kindergartens in eight cities across six major administrative regions by stratified cluster sampling, and the P-PAQ initially developed by the researchers of this study were completed by the primary caregivers. The questionnaire was administered to collect data relating to the amount of physical activity undertaken by the preschoolers, and the norm was determined by quartiles. Data relating to parental concepts of sports and parental behavior were assessed by calculating mean scores in order to establish the norm. Results: Among preschoolers in urban areas, the M(P 25 ，P 75 ) of total physical activity time (min/day), moderate-to-vigorous physical activity time (min/day), outdoor time (min/day) and screen time (min/day) on school days outside kindergarten and on weekends were 84 (54,120), 22 (8,40), 12 (0,24) and 18 (6,30), and 170 (115,240), 60 (30,95), 90 (35,120) and 30 (20,60), respectively. When the score of parents sports concept and behavior (total score of 40) were≥34, 29-<34, 24-<29, <24, it was defined as four levels about above medium, medium, lower medium and lower, respectively. And for two dimensions，when the score of parental sports concept were ≥19, 17-<19, 15-<17, <15，and the score of parental behaviors were ≥16, 12-<16, 8-<12, <8, it was defined as four levels about upper medium, medium, lower medium and lower, respectively. Conclusion The norm of extracurricular activities among preschool children in Chinese cities has good representativeness and appropriate threshold values, which could provide a valuable reference for early assessment, as well as guidance in relation to out-of-school physical activity behaviors among children aged 3-6 years old.

Drugs and physical stimulation,including light,electricity,and magnetic fields,can be used to influence how neurons operate,among which chemogenetic and optogenetic technologies are most widely used.In recent years,magnetogenetic technology has also been developed that can acti-vate neurons in magnetic fields through magnetic sensitive actuators,leading to non-invasive and instanta-neous activation of specific brain regions.This article reviews the evolution of and problems with chemoge-netic and optogenetic techniques commonly used in brain science research.It also outlines the latest progress in magnetogenetic technologies,which are not full-fledged yet,as well as the role of transcra-nial electrical stimulation,transcranial magnetic stimulation,deep brain stimulation and transcranial ultra-sound stimulation technology in the functional regulation of brain diseases.Constant adjustment and improvement can make it possible for these technologies to be used more widely for the study of brain sciences and the treatment of brain diseases.

Pathogenic microorganism is an important national strategic resource, whose value is embodied in the physical object and its corresponding information and data resources. At present, China has basically completed the designation of national and provincial preservation centers and professional laboratories, and various preservation institutions have been put into operation and begun to play its role of preservation institutions. In order to standardize the data management of pathogenic microorganism preservation and improve the quality of pathogenic microorganism resources, China CDC took the lead in formulating and issuing the community standard of Chinese Preventive Medicine Association, the General principles of description for pathogenic microorganism collection (T/CPMA 011-2020). The standard puts forward the data fields and description principles of microorganism strain, including general data such as number, name, separation, hazard classification, transmission route, pathogenicity, and characteristic data such as virus, bacteria and fungi. The core of pathogenic microorganism resources lies in quality and the foundation lies in standards. Taking data standards as the starting point, it will play an important supporting role in promoting the transformation of pathogenic microorganism preservation work to quality improvement, improving resource sharing and utilization, and leading the sustainable development of preservation work.

Objective To analyze the expression of CREM in gastric cancer(GC)and its correlation with prognosis of the patients.Methods TCGA and GEO databases were used to analyze the expression levels of CREM mRNA in GC and adjacent tissues.Immunohistochemistry was used to examine the expression of CREM protein in 43 pairs of GC and adjacent tissues,and the correlation of CREM expression with clinicopathological features of the patients was analyzed.Kaplan-Meier survival analysis was used to explore the relationship between CREM expression and survival of GC patients.LinkedOmics database was used to annotate the GO function and KEGG pathway enrichment of CREM-related genes.Results Database analysis showed that CREM was highly expressed in GC tissues(P<0.05)and positively correlated with poor prognosis in GC patients(P=0.01).Immunohistochemistry results showed significantly higher CREM expression in GC tissues than in paired adjacent tissues(P<0.0001),and its expression level was correlated with T-stage and N-stage of the tumor(P<0.05).The overall survival of GC patients with high expression of CREM was shorter(RR=4.02,P=0.0046).Gene enrichment analysis showed that high CREM expression promotes occurrence and progression of GC very likely through the cell adhesion signaling pathway.Conclusion CREM is highly expressed in GC,and its high expression is associated with a poor prognosis of GC patients,suggesting the potential of CREM to serve as a prognostic indicator for GC.

Objective To analyze the expression of CREM in gastric cancer(GC)and its correlation with prognosis of the patients.Methods TCGA and GEO databases were used to analyze the expression levels of CREM mRNA in GC and adjacent tissues.Immunohistochemistry was used to examine the expression of CREM protein in 43 pairs of GC and adjacent tissues,and the correlation of CREM expression with clinicopathological features of the patients was analyzed.Kaplan-Meier survival analysis was used to explore the relationship between CREM expression and survival of GC patients.LinkedOmics database was used to annotate the GO function and KEGG pathway enrichment of CREM-related genes.Results Database analysis showed that CREM was highly expressed in GC tissues(P<0.05)and positively correlated with poor prognosis in GC patients(P=0.01).Immunohistochemistry results showed significantly higher CREM expression in GC tissues than in paired adjacent tissues(P<0.0001),and its expression level was correlated with T-stage and N-stage of the tumor(P<0.05).The overall survival of GC patients with high expression of CREM was shorter(RR=4.02,P=0.0046).Gene enrichment analysis showed that high CREM expression promotes occurrence and progression of GC very likely through the cell adhesion signaling pathway.Conclusion CREM is highly expressed in GC,and its high expression is associated with a poor prognosis of GC patients,suggesting the potential of CREM to serve as a prognostic indicator for GC.

ObjectiveTo investigate the dietary preference and nutritional knowledge needs of the elderly people who dined at meal service sites. MethodsUsing the form of stratified and convenience sampling method with self-designed questionnaire was used， in November 2021， to select 700 elderly people who dine at meal service sites in 7 jurisdictions in Shanghai were selected， and a self-designed questionnaire was used to investigate the basic information. Results91.64% of the elderly surveyed would eat at relatively fixed meal service sites， and the total Dietary Diversity Score （DDS9） was 3.56±1.46. 41.45% of the elderly with diseases preferred unhealthy cooking methods. Only 8.03% of the surveyed seniors said they were unwilling to accept targeted and personalized nutrition tips and reminders. Multivariate logistic regression analysis showed that the probability reaching the “understanding” level of “Food Guide Pagoda for Chinese Residents” and “Four Principles Recommended by the Core Dietary Guidelines for the Elderly” was different in the elderly with different education levels. The willingness of the elderly to expect to receive different nutrition tips and reminders was related to whether they cared about the corresponding contents. There was a statistically significant difference （P<0.05） among the elderly who were concerned about different health problems in terms of the willingness to receive different nutritional tips. There were significant differences in the proportion of elderly people with different health status for intervention （χ²=5.402， P<0.05）. ConclusionThe elderly who dine at meal service sites are highly dependent on the sites， have a low level of dietary diversification， and do not have a high degree of understanding of nutrition-related knowledge， and have a high demand for targeted nutritional interventions. Nutritional interventions for the sick elderly should be piloted through multiple channels.