Objective:To investigate the application effect of research-assisted teaching mode in histology teaching by taking the construction of rat spinal cord injury model as an example.Methods:A convenient sampling method was used to select 52 freshmen majoring in clinical medicine of grade 2020 in our school as the research subjects.They were randomly divided into the conventional teaching group and the research-assisted teaching group,with 26 students in each group.The conventional teaching group received traditional classroom teaching combined with experimental teaching.On this basis,research elements were added to the experimental teaching in the research-assisted teaching group to provide more in-depth research methods and experimental design,so as to cultivate students'research abilities.The theoretical assessment and practical skills assessment of histology course were compared between the two groups.A subjective evaluation questionnaire on experimental animal models teaching was used to evaluate the advantages of teaching programs in three dimensions:psychological quality training,technical skill enhancement,and theoretical knowledge expansion.The research abilities of the two groups were compared.Pearson correlation analysis was used to analyze the correlation between research ability scores and comprehensive scores.Results:The usual score and comprehensive score in the research-assisted teaching group were significantly higher than those in the conventional teaching group(P＜0.05).The scores of adaptability to challenging tasks,determination and perseverance in problem-solving,communication skills in teamwork,literature reading,integration of practical experience with theoretical knowledge,practical application value of the course,and innovation of teaching resources in the research-assisted teaching group were higher than those in the conventional teaching group(P＜0.05).The scores in experimental design ability,background knowledge in literature review,communication and reporting skills,awareness of evidence-based medicine,and overall research ability in the research-assisted teaching group were higher than those in the conventional teaching group(P＜0.05).The total score of students'research abilities was positively correlated with comprehensive scores(r=0.716,P＜0.01).Conclusions:The application of research-assisted teaching significantly improved students'research abilities in histology teaching.The enhancement of research abilities can promote the mastery of theoretical knowledge and the improvement of experimental skills.

OBJECTIVE To study the inhibitory effect and mechanism of curcumin and its derivatives A and B on TGF-β induced LX-2 cell fibrosis. METHODS Established the liver fibrosis model of LX-2 cells induced by TGF-β(10 ng·mL−1).The effects on cell proliferation were detected by CCK-8. The effects on cell apoptosis was detected by flow cytometry. The effects on fibrosis related factors(Collagen I, Collagen Ⅳ, Fibronectin, Vimentin, α-SMA, PDGFRβ, TGFβR1, TGFβR2, MMP2, MMP9, TIMP1 and TIMP2) protein expression and gene transcription levels were detected by Western blotting and q-PCR. RESULTS The curcumin and its derivative A and B had the inhibition effects on normal LX-2 cells, and the IC25 values were 15.7, 2.6, 10.2 μmol·L−1, respectively. Compared to the model group, the curcumin(15.7 μmol·L−1) and its derivative A(2.6 μmol·L−1) and B(10.2 μmol·L−1) had the significant inhibition effects on cell proliferation of the TGF-β induced LX-2 cells(P<0.05). The cell apoptosis rate of curcumin derivative B group was higher than the model group(P<0.05). Collagen I, Fibronectin, Vimentin, α-SMA, TGFβR1 and TIMP-1 protein expression levels in curcumin group were lower, while the protein expression level of MMP-9 was higher(P<0.05). The protein expression levels of Collagen I, Collagen IV, Fibronectin, Vimentin, α-SMA, TIMP-1 and TIMP-2 in curcumin derivative A group were lower, while the protein expression level of MMP-2 was higher(P<0.05). The protein expression levels of Collagen I, Collagen IV, Fibronectin, Vimentin, α-SMA, PDGFRβ, TGFβR1, TGFβR2, TIMP-1 and TIMP-2 in curcumin derivative B group were lower, while the protein expression level of MMP-2 was higher(P<0.05). The gene transcription levels of Collagen I, Fibronectin, α-SMA and TIMP-1 in curcumin group were lower(P<0.05). The gene transcription levels of Collagen I, Fibronectin and α-SMA in curcumin derivative A and B groups were lower(P<0.05). CONCLUSION Curcumin and its derivatives A and B inhibit the abnormal activation and proliferation of TGF-β-induced LX-2 cells, inhibit the excessive secretion and accumulation of its extracellular matrix components, and promote its degradation, thus playing an anti-fibrotic effect in vitro, especially the curcumin derivative B.

OBJECTIVE To obtain stronger cytotoxic activity of panaxadiol derivatives. METHODS The 3-amino panaxadiol was prepared by the bioelectronic isosteric principle, and then 18 derivatives of cinnamic acid, NO donor and other types of panaxadiol derivatives were synthesized, among them, 12 compounds had not been reported in the literature, and their structures had been confirmed by 1H-NMR, 13C-NMR and mass spectrometry. These compounds were evaluated for their cytotoxic activity by MTS assay against human leukemia cell line HL-60, liver cancer cell line SMMC-7721, lung cancer cell line A-549, breast cancer cell line MCF-7, and colon cancer cell line SW480. RESULTS These results showed that compounds 6c, 7 as well as 7j exhibited potent inhibitory activities against all five tumor cells, especially the IC50 values of compound 7 against HL-60 and SMMC-7721cells were 3.41 and 4.51 μmol·L−1, respectively. It was significantly superior to panaxadiol in cytotoxicity. CONCLUSION These results show that 7 and 7j can be used as promising lead compounds for further research.

Pulmonary blast injury has become the main type of trauma in modern warfare, characterized by externally mild injuries but internally severe injuries, rapid disease progression, and a high rate of early death. The injury is complicated in clinical practice, often with multiple and compound injuries. Currently, there is a lack of effective protective materials, accurate injury detection instrument and portable monitoring and transportation equipment, standardized clinical treatment guidelines in various medical centers, and evidence-based guidelines at home and abroad, resulting in a high mortality in clinlcal practice. Therefore, the Trauma Branch of Chinese Medical Association and the Editorial Committee of Chinese Journal of Trauma organized military and civilian experts in related fields such as thoracic surgery and traumatic surgery to jointly develop the Clinical treatment guideline for pulmonary blast injury ( version 2023) by combining evidence for effectiveness and clinical first-line treatment experience. This guideline provided 16 recommended opinions surrounding definition, characteristics, pre-hospital diagnosis and treatment, and in-hospital treatment of pulmonary blast injury, hoping to provide a basis for the clinical treatment in hospitals at different levels.

Abstract OBJECTIVE To study the mechanism of NLRP3 gene’s regulation on inflammatory response in non-alcoholic fatty liver disease(NAFLD) mice induced by high-fat and high-fructose diet using NLRP3 gene knockout mice. METHODS Use male homozygous(NLRP3-/-) mice, and the high-fat and high-fructose diet was used to establish NAFLD model in NLRP3 knockout(KO) mice and wild-type(WT) mice, divided into KO high-fat and high-fructose diet(KO-HFD) group and WT high-fat and high-fructose diet(WT-HFD) group, while the WT and KO groups were also established. The body weight of mice in each group were observed. The changes of ALT, AST, TG, TC, MDA, SOD, lipidosis, apoptosis rate, IL-1β, IL-18, TNF-α, NF-κB, NLRP3, Caspase-1 and ASC in serum and tissue samples were tested to study the mechanism of the NLRP3 gene regulating the inflammatory response in NAFLD mice. RESULTS As time goes on, the mice weight of each group increased gradually, but the KO-HFD group increased less than the WT-HFD group. Each group’s level of ALT, AST, TG, TC in serum increased gradually, but the KO-HFD group increased less than the WT-HFD group. The level of MDA in liver tissues of each group was gradually increased and the level of SOD was gradually decreased, but the change range of KO-HFD group was smaller than that of WT-HFD group. The results of oil red O staining and Tunel section showed that the degree of lipid deposition and apoptosis in hepatocytes increased gradually in all groups, but the changes in KO-HFD group were less than that in WT-HFD group. The levels of serum and liver inflammatory factors IL-1β, IL-18, TNF-α and NF-κB increased in all groups, but the changes of KO-HFD group 20 weeks were less than those of WT-HFD group 20 weeks, and the difference was statistically significant. The expression levels of NLRP3 inflammasomes and related inflammatory cytokines NLRP3, Caspase-1, ASC, IL-1β and IL-18 in liver tissues of WT-HFD group were higher than those of KO-HFD group. The mRNA transcription levels of NLRP3, ASC, Caspase-1, IL-1β and IL-18 in WT-HFD group were gradually increased, while there was no change in KO-HFD group. CONCLUSION The NLRP3 gene may be activated in NAFLD mice model, resulting in increased expression of NLRP3 inflammasome-associated protein, promoting the synthesis and secretion of downstream inflammatory factors, resulting in significant inflammatory response and liver damage, and promoting the progression of NAFLD disease.

Objective: To create a model for early prediction of essential hypertension (EH) based on the TreeNet algorithm, so as to provide a tool for early monitoring of EH. Methods: The health examination data were collected from individuals receiving health examinations in Hangzhou Haiqin Health Examination Center or Shanghai Yibao Health Management Co., Ltd from 2014 to 2016, and a predictive model for EH was created based on the TreeNet algorithm. The effectiveness of the model for early prediction of EH was evaluated using root mean square error (RMSE), mean absolute deviation (MAD), coefficient of determination (R2) and receiver operating characteristic (ROC) curve. Results: A total of 12 variables were included in the model, and the highest contributing variable was body mass index (BMI), followed by BMI difference, two-year BMI difference, two-year triglyceride (TG) difference, two-year total cholesterol (TC) difference, high-density lipoprotein cholesterol (HDL-C) in 2014, TG in 2014, low-density lipoprotein cholesterol (LDL-C) in 2014, body weight in 2015, fasting blood glucose in 2015, TG in 2015, urea nitrogen difference and platelet in 2015. The highest predictive accuracy was 100.00%, and the lowest was 56.89%. The risk of EH significantly increased among individuals with BMI in 2015 of >25 kg/m2, two-year BMI difference of >0.5 kg/m2, two-year TG difference ranging from 1.3 to 3.3 mmol/L, TC in 2015 of 2.0 to 2.4 mmol/L and HDL-C in 2014 of <0.52 mmol/L. The model presented RMSE of 0.082, MAD of 0.064, R2 of 0.811, area under the ROC curve of 0.788 (95%CI: 0.741-0.815), sensitivity of 69.05% and specificity of 66.21% for prediction of EH Conclusion The TreeNet algorithm-based model is effective for early monitoring of high-risk individuals for EH.

Objective:To analyze the risk factors of bronchopulmonary dysplasia(BPD)in very preterm infants(VPI), and to provide scientific basis for the prevention and treatment of BPD in VPI.Methods:A prospective multicenter study was designed to collect the clinical data of VPI in department of neonatology of 28 hospitals in 7 regions from September 2019 to December 2020.According to the continuous oxygen dependence at 28 days after birth, VPI were divided into non BPD group and BPD group, and the risk factors of BPD in VPI were analyzed.Results:A total of 2 514 cases of VPI including 1 364 cases without BPD and 1 150 cases with BPD were enrolled.The incidence of BPD was 45.7%.The smaller the gestational age and weight, the higher the incidence of BPD( P<0.001). Compared with non BPD group, the average birth age, weight and cesarean section rate in BPD group were lower, and the incidence of male infants, small for gestational age and 5-minute apgar score≤7 were higher( P<0.01). In BPD group, the incidences of neonatal respiratory distress syndrome(NRDS), hemodynamically significant patent ductus arteriosus, retinopathy of prematurity, feeding intolerance, extrauterine growth restriction, grade Ⅲ~Ⅳ intracranial hemorrhage, anemia, early-onset and late-onset sepsis, nosocomial infection, parenteral nutrition-associated cholestasis were higher( P<0.05), the use of pulmonary surfactant(PS), postnatal hormone exposure, anemia and blood transfusion were also higher, and the time of invasive and non-invasive mechanical ventilation, oxygen use and total hospital stay were longer( P<0.001). The time of starting enteral nutrition, cumulative fasting days, days of reaching total enteral nutrition, days of continuous parenteral nutrition, days of reaching 110 kcal/(kg·d) total calorie, days of reaching 110 kcal/(kg·d) oral calorie were longer and the breastfeeding rate was lower in BPD group than those in non BPD group( P<0.001). The cumulative doses of amino acid and fat emulsion during the first week of hospitalization were higher in BPD group( P<0.001). Multivariate Logistic regression analysis showed that NRDS, invasive mechanical ventilation, age of reaching total enteral nutrition, anemia and blood transfusion were the independent risk factors for BPD in VPI, and older gestational age was the protective factor for BPD. Conclusion:Strengthening perinatal management, avoiding premature delivery and severe NRDS, shortening the time of invasive mechanical ventilation, paying attention to enteral nutrition management, reaching whole intestinal feeding as soon as possible, and strictly mastering the indications of blood transfusion are very important to reduce the incidence of BPD in VPI.

Boao Lecheng International Medical Tourism Pilot Area is the only administrative area in China where the pilot application of real-world data is performed. Based on the special healthcare policy in Boao Lecheng, conducting real-world data study to provide real-world evidence for the clinical evaluations and approvals of innovated medical products has become an important field that supports the reform of China's evaluation and approval system of medical products. Trustworthy real-world evidence needs to rely on both high-quality real-world data and reasonable and rigorous study designs. Based on the real-world data research guidelines and specifications issued by relevant academic research and regulatory authorities both at home and abroad, combined with the special policy environment of Boao Lecheng and previous practice experience, this paper summarizes the study designs of real world data in Boao Lecheng and discusses the key considerations of different study design in the context of special healthcare policies in Boao Lecheng in order to provide reference for the further study of real-world data.

Objective To investigate the functions of microRNA-1 43 (miR-1 43)in esophageal cancer cell line ECA1 09.Methods ECA1 09 cells were transfected with negative control (NC),miR-1 43 mimics or miR-1 43 inhibitors.3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT)assay was per-formed to evaluate the growth of ECA1 09 cells after transfection.Annexin V-FITC /PI apoptosis test kit was used to detect early apoptosis rate in ECA1 09 cells.Transwell migration and invasion assays were conducted to compare the migration and invasion capacity of ECA1 09 among different groups.Real-time PCR and Western blotting were used to analyze the mRNA and protein alteration after transfection.Results Three and four days after transfection,compared with NC (absorbance value:0.90 ±0.02 and 1 .09 ±0.07),miR-1 43 mimics inhibited ECA1 09 cell proliferation (absorbance value:0.66 ±0.05 and 0.80 ±0.04),while miR-1 43 inhibi-tors promoted cell proliferation (absorbance value:1 .1 3 ±0.09 and 1 .51 ±0.08),with statistical signifi-cances (F =49.1 6,P =0.000;F =1 00.34,P =0.000).Early-stage apoptosis rates of ECA1 09 transfected with NC,miR-1 43 mimics and miR-1 43 inhibitors were 3.42% ±0.72%,1 1 .63% ±1 .1 5% and 0.94% ± 0.1 0%,respectively,with statistical significance (F =1 51 .61 ,P =0.000).Meanwhile,compared with NC (migration cell number:336 ±1 3,invasion cell number:1 47 ±1 6),miR-1 43 mimics inhibited cell migration (1 48 ±1 6)and invasion (75 ±1 0),while miR-1 43 inhibitors promoted cell migration (51 0 ±1 4)and inva-sion (238 ±1 6),with statistical significances (F =470.99,P =0.000;F =90.04,P =0.000).Compared with NC (1 .00 ±0.00),miR-1 43 mimics down-regulated mRNA (relative expression level 0.22 ±0.08)and protein expression (relative expression level 0.46 ±0.08)of K-ras,whereas miR-1 43 inhibitors up-regulated mRNA (1 .55 ±0.1 2)and protein expression (1 .33 ±0.05)of K-ras (F =1 31 .36,P =0.000;F =88.1 7, P =0.000).Conclusion miR-1 43 functions as a tumor suppressor in esophageal cancer cell line ECA1 09, probably by down-regulating K-ras expression.

OBJECTIVETo investigate the relationship between the expression of hepatitis B virus (HBV) core antigen and viral replication and liver tissue inflammation damage in chronic hepatitis B (CHB) patients, and to analyze the relationship of core antigen expression differences with clinical and pathological features in e antigen-negative and e antigen-positive CHB patients.

METHODSSixty-three treatment-naive patients diagnosed with CHB who underwent liver biopsy were included in this retrospective analysis. Liver pathology was assessed, and the karyotype, pulp type, and pulp karyotype were determined. Core and e antigen expression was quantitatively determined by automated immunoassay. Blood samples were used to determine the amount of peripheral lymphocytes or monocytes and HBV DNA load. Results The median titer of HBV DNA was significantly higher in the CHB patients with e antigen positivity (n = 48) than those with e antigen negativity (n = 15) (5.4 * 106 copies/ml vs. 5.4 * 104 copies/ml, P = 0.003). The core antigen positive expression rate was significantly higher in the e antigen-positive CHB patients than in the e antigen-negative CHB patients (80.33% vs. 53.33%, P = 0.042). For the e antigen-positive CHB patients, the HBV DNA titer in karyotype core antigen cases was higher than that in the pulp karyotype mixed-type cases (P = 0.008) and in the negative cases (P = 0.013); in addition, the karyotype patients showed higher titer than the plasma patients (P = 0.019). Also for the e antigen-positive CHB patients, the HBV DNA titer was positively correlated with the rank level of pulp karyotype in core antigen expression (r = 0.589, P = 0.003) but negatively correlated with lobular inflammation, interface inflammation, and fibrosis level (r = -0.552, P = 0.000; r = -0.381, P = 0.008; r = -0.555, P = 0.000); in addition, the level of peripheral blood lymphocytes was negatively correlated with lobular inflammation and fibrosis level (r = -0.361, P = 0.012; r = -0.356, P = 0.013). For the e antigen-negative CHB patients, the level of peripheral blood lymphocytes was negatively correlated with lobular inflammation and interface inflammation (r = -0.702, P = 0.004; r = -0.578, P = 0.024), while the level of peripheral blood mononuclear cells was negatively correlated with lobular inflammation, interface inflammation, and fibrosis level (r = -0.682, P = 0.005; r = -0.620, P = 0.014; r = -0.527, P = 0.044); in addition, age positively correlated with interface inflammation (r = 0.690, P = 0.004).

CONCLUSIONThe pulp karyotype mixed-type of core antigen expression may reflect the level of HBV replication. Negative expression of core antigen may be associated with variation in pre-C or C zone. The monocyte-macrophage system may be involved in the pathogenesis of e antigen-negative CHB, while the mechanism of immune escape may play an important role in increasing HBV DNA titer in an e-antigen-negative CHB condition.

Adult ; Aged ; DNA, Viral ; blood ; Female ; Hepatitis B Core Antigens ; blood ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; blood ; pathology ; virology ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Viral Load ; Virus Replication ; Young Adult