[Application and advancement of digital teaching materials in teaching epidemiology].

This study aims at examining the application and development of digital teaching materials in the field of epidemiology, encompassing both China and international contexts. The research involved conducting search on websites and literature databases to assess the status of digital teaching materials in epidemiology, nationally and internationally. At present, in China, digital teaching materials used in epidemiology are primarily presented in the form of printed books with added QR codes, providing teaching resources such as videos and exercises. However, issues with the level of interactivity have been identified. In foreign countries, with stronger emphasis placed on personalization, interactivity, and the use of rich media technologies in the digital teaching materials, epidemiologically. Enhanced digitization regarding materials and learning outcomes is achieved through features such as real-time notes, interactive animations, and quizzes. These approaches are considered worth considering for adoption. This study provides valuable insights for the digital transformation of epidemiology education.

[Research progress of cell pyroptosis in the mechanism and treatment of glaucoma].

Pyroptosis, which is put forward after apoptosis, necrosis and autophagy, is a programmed pro-inflammatory cell death mode mainly mediated by inflammasomes. The main signaling pathways include the classic caspase-1-dependent pyroptosis pathway and the non-classical caspase-4, caspase-5 and caspase-11-dependent pyroptosis pathway. Pyroptosis is involved in the normal physiological function and plays an important role in the pathogenesis of many diseases, especially in neurodegenerative diseases such as Alzheimer's disease and glaucoma. Glaucoma, which is mainly related to pathological intraocular pressure elevation, is a kind of eye diseases that threaten and damage the optic nerve and its visual pathway and eventually lead to visual function impairment. With more attention paid to pyroptosis involved in the mechanism of eye diseases and the pathogenesis and treatment of glaucoma, studies have found that pyroptosis is tightly correlated with glaucoma. Pyroptosis of retinal ganglion cells and microglia-mediated pyroptosis possess a significant position in the development of glaucoma. Further understanding of the relationship between pyroptosis and glaucoma might bring a new dawn for glaucoma treatment. The mechanism of pyroptosis may provide a new strategy for the pathogenesis and treatment of glaucoma. (Chin J Ophthalmol, 2021, 57: 702-706).

[Abnormal expression of Wnt3a and inhibiting role of its molecular-targeted intervening in hepatocellular carcinoma].

Objective: To investigate the Wnt3a expression in tissues of HCC and its gene knockout on effects of HepG2 cell proliferation or xenograft tumor growth. Methods: Hepatic Wnt3a expressions in 87 HCC and their matched surrounding tissues were observed by tissue microarray and immunohistochemistry for analyzing its clinicopathological characteristics; Wnt3a-knockout HepG2 cell lines were established by Crispr/cas9-sgRNA system and genomic cleavage efficiency was verified at gene level by surveyor assay. The relative proteins were confirmed by Western blotting; Cell Counting Kit-8 assay was used to examine cell proliferation after knocking-out Wnt3a successfully, and the nude mice HepG2 cell xenograft tumors delete that the relationship between Wnt3a and HCC growth. Results: The positive Wnt3a with brown staining particles was mainly distributed in cytosol and membrane of hepatocytes. The incidence of hepatic Wnt3a expression in cancerous tissues (95.4%) was significantly higher (χ (2) = 47.754, P < 0.001) than that in their surrounding tissues (49.4%). The high Wnt3a expression was 70.1% in the HCC and only 14.9% in the surrounding tissues. High Wnt3a expression was associated with poorly-differentiated grade, liver cirrhosis, HBV infection, portal vein invasion, TNM stage and 5-year survival rate. After knocked-out by Crispr/cas9-sgRNA system successfully, Wnt3a expression was down-regulated significantly at gene or protein level. Key molecule ß-catenin in cytoplasma was obviously inhibited. HepG2 cell lines proliferation was suppressed in time-dependent manner. The nude mice HepG2 cell xenograft tumors confirmed that the knock-out of Wnt3a could significantly supressed HCC growth with slower speed (t = 6.418, P < 0.001), smaller volume(869.4 ± 222.5 mm(3) vs 355.0 ± 99.9 mm(3), t = 5.168, P < 0.001), and lighter weight (0.88 ± 0.20 g vs 0.35 ± 0.11 g, t = 5.628, P < 0.001)compared with the control group. Conclusion: Abnormal expression of Wnt3a could be expected as a promising target for HCC gene therapy.

[Abnormal expression of angiopoietin-2 associated with invasion, metastasis and prognosis of lung cancer].

Objective: To investigate the relationship between abnormal angiopoietin-2 (Ang-2) expression and invasion/metastasis of lung cancer. Methods: Totally 122 cases of postoperative primary lung cancer tissues and their paracancerous tissues from Jan. 2009 to Dec. 2010 were collected from Affiliated Hospital of Nantong University and Ang-2 expression was analyzed by immunohistochemistry. At the cellular level, the protein and mRNA levels of Ang-2 in lung epithelial cell line Beas-2B and four lung cancer cell lines (SPCA-1, NCI-1650, A549 and NCI-H1975) were observed. The most effective Ang-2-shRNA for Ang-2 transcription was screened and transfected into A 549 lung cancer cells. The Ang-2 expression, Ang-2 gene transcription, cell proliferation, invasion/metastasis, and epithelial-mesenchymal transition (EMT) abilities of lung cancer cells were analyzed by Western blotting, fluorescent quantitative reverse transcriptase PCR, Cell Counting Kit-8 assay, and Transwell cell models for exploring the relationship between Ang-2 expression and invasion/metastasis of lung cancer. Results: The higher Ang-2 expression levels in lung cancerous tissues were closely related to tumor diameter (P=0.008), differentiating degree (P=0.033), TNM stage (P=0.025) and 5-year survival rate (P<0.001). According to the Kaplan-Meier survival curves, the 5-year survival rate of patients with higher expression levels of Ang-2 (16.1%) was significantly poorer than that of patients with lower Ang-2 (80.0%, P<0.001). Significant difference of 5-year survival rate was found in patients with different Ang-2 levels at TNM stage Ⅰ(P<0.001), but not at stage Ⅱ, Ⅲ and Ⅳ. Among Beas-2B and four lung cancer cell lines, the protein and mRNA levels of Ang-2 in A549 cells were the highest. After Ang-2-shRNA-1 plasmid successfully transfected into A549 cells, cell proliferation rate was significantly lower than that in the shRNA-negative or blank group at a time-dependent manner. The significant decrease of the invasion, migration and EMT abilities were also found in A549 cells after transfection of Ang-2 shRNA. Conclusion: Abnormal expression of Ang-2 is closely related to invasion, migration and prognosis of lung cancer, and interfering the activation of Ang-2 would be a novel molecular-targeted therapy for lung cancer.

[Clinical application of high mobility group box nuclear protein 1 subcomponent in the progression of liver cancer].

The biological functions of high-mobility group (HMG) proteins include regulation of DNA replication, transcription, recombination and repair. According to molecular weight, sequence alignment and DNA structural characteristics, HMG proteins are subdivided into three superfamilies (HMGA, HMGB and HMGN). Recently, HMGB family members (HMGB1, HMGB2, HMGB3, and HMGB4) found to interact with hepatitis B or C virus. Therefore, activation of relevant signaling molecules to regulate transcription of genes related to hepatocellular carcinoma as a mediator of inflammation promoting HCC progression has attracted considerable attentions. This article focuses on the clinical application of the expression of HMGB family members in the process of HCC progression.

[Molecular markers for the monitoring of malignant transformation of hepatocytes and early diagnosis of liver cancer].

The monitoring of malignant transformation of hepatocytes or early diagnosis of primary hepatocellular carcinoma (PHC) remains a challenge in the medical world. Routine examinations including serum alpha-fetoprotein level and ultrasound examination have a limited value in the diagnosis of small hepatocellular carcinoma; however, the effective treatment of PHC depends on its early diagnosis. In recent years, molecular markers including important signaling molecules in PHC-related pathways, carcinoembryonic proteins, and non-coding RNA help with the monitoring of malignant transformation of hepatocytes or early diagnosis of liver cancer. This article reviews the valuable molecular markers in the monitoring of malignant transformation of hepatocytes or early diagnosis of liver cancer.

[Dynamic expression of carnitine palmitoyltransferase II in the mitochondrial inner membrane during hepatocyte malignant transformation induced by lipid accumulation].

Objective: To investigate the dynamic expression of hepatic carnitine palmitoyltransferase-II (CPT-II) in the mitochondrial inner membrane during hepatocyte malignant transformation induced by lipid accumulation. Methods: Male Sprague-Dawley rats were divided randomly into control, fatty liver, and induced cancer groups, which were fed with normal, high-fat (HF), and HF containing 2-fluorenylacetamide (0.05%, 2-FAA) diets, respectively, for 14 weeks. One rat from each group was sacrificed every two weeks and the blood and liver samples were collected. Liver morphological changes were evaluated with hematoxylin and eosin staining, and the liver tissue samples were divided into control, fatty liver, degeneration, precancerous, and cancerous groups accordingly. Hepatic lipids were dyed by the oil red O method. The CPT-II expression was measured by immunohistochemistry and compared with the specific CPT-II concentration (ng/mg liver protein, ng/mg P) among different groups. Serum levels of circulating total cholesterol (Tch), triglyceride (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were quantitatively analyzed. Results: Massive lipid accumulation hepatocytes was seen in rats on HF and HF containing 2-FAA diets. The lipid levels in the control group were significantly lower than those in the fatty liver (t = -11.556, P < 0.001), degeneration (t = -4.847, P = 0.04), precancerous (t = -13.652, P = 0.005), and cancerous groups (t = -10.896, P = 0.008). The serum TG and Tch levels in the degeneration, precancerous, and cancerous groups were 2-3 times higher than those in the control group (P < 0.05). After 2-FAA treatment, the morphological changes of rat hepatocytes showed the progression from degeneration and precancerosis to cancerosis, with hepatocyte injury. The serum AST and ALT levels in the degeneration, precancerous, and cancerous groups were significantly higher (4-8 times) than those in the control group (P < 0.05). The specific concentration of liver CPT-II expression was significantly reduced during hepatocyte malignant transformation, as confirmed by immunohistochemistry, with the CPT-II levels significantly lower in the cancerous group than in any of other groups (P < 0.05). Conclusion: Low hepatic CPT-II expression might lead to abnormal lipid accumulation in hepatocytes, which should promote the malignant transformation of hepatocytes.

[Advances in basic and clinical research on hepatocellular carcinoma in 2016].

Hepatocellular carcinoma (HCC) is still one of common malignant cancers worldwide, with increasing incidence and mortality rates. Early diagnosis and effective treatment for HCC remain to be explored. This article introduces the research advances in the early specific diagnosis and effective therapies for HCC in 2016, such as molecular markers in the specific diagnosis and targeted therapy for HCC, main therapeutic regimens, robot-assisted liver resection, and no-touch radiofrequency ablation.

[Clinical values of Wnt3a as a novel biomarker in diagnosis and prognosis of hepatocellular carcinoma].

Objective: To explore Wnt3a expression in HCC tissues and serum, and to discuss its clinical diagnostic and prognostic value. Methods: The Wnt3a expressions were detected in a total of 186 patients (HCC, liver cirrhosis and chronic Hepatitis) and 40 controls by Elisa, comparing with AFP to evaluate its clinical diagnosis value. Wnt3a expressions in 80 HCC and surrounding tissues were analyzed by IHC, to explore its prognostic value. Results: Wnt3a with brown staining was mainly distributed in cytosol and of hepatocyte membrane. The higher expression (3-6 scores) was 71.3% in HCC, 13.8% in surrounding tissues, associated with poorly-differentiated grade, liver cirrhosis, HBV infection, higher TNM stage (P<0.05) and 5-year survival rate (P<0.001), identified as independent predictive factors for poor HCC outcome and closely related with lower five-year survival rate. Serum average Wnt3a levels were significantly higher (P<0.001) in the HCC group than those in any other groups of benign liver diseases, with about 4.0, 9.2 and 26.7 times higher than that in the liver cirrhosis, chronic hepatitis and normal control group. Wnt3a expression in HCC were closely related to AFP concentration, liver cirrhosis HBV infection, poor differentiation, TNM stagingand extra- hepatic metastasis (P<0.05). The sensitivity, specificity, accuracy, positive predictive value and negative predictive values were 92.5, 94.3, 93.2, 96.1 and 89.3% at 800 ng/L as cutoff value for Wnt3a. Combining Wnt3a and AFP test, the total sensitivity could rise to 96.3%. The area under ROC curve in Wnt3a (0.994)was higher than in AFP (0.710). Conclusions: Wnt3a as a critical signal molecule in the Wnt pathway is a new specific marker for HCC diagnosis and prognosis.

Long-term treadmill exercise improves spatial memory of male APPswe/PS1dE9 mice by regulation of BDNF expression and microglia activation.

Increasing evidence suggests that physical activity could delay or attenuate the symptoms of Alzheimer's disease (AD). But the underlying mechanisms are still not fully understood. To investigate the effect of long-term treadmill exercise on the spatial memory of AD mice and the possible role of ß-amyloid, brain-derived neurotrophic factor (BDNF) and microglia in the effect, male APPswe/PS1dE9 AD mice aged 4 months were subjected to treadmill exercise for 5 months with 6 sessions per week and gradually increased load. A Morris water maze was used to evaluate the spatial memory. Expression levels of ß-amyloid, BDNF and Iba-1 (a microglia marker) in brain tissue were detected by immunohistochemistry. Sedentary AD mice and wildtype C57BL/6J mice served as controls. The results showed that 5-month treadmill exercise significantly decreased the escape latencies (P < 0.01 on the 4th day) and improved the spatial memory of the AD mice in the water maze test. Meanwhile, treadmill exercise significantly increased the number of BDNF-positive cells and decreased the ratios of activated microglia in both the cerebral cortex and the hippocampus. However, treadmill exercise did not significantly alleviate the accumulation of ß-amyloid in either the cerebral cortex or the hippocampus of the AD mice (P > 0.05). The study suggested that long-term treadmill exercise could improve the spatial memory of the male APPswe/PS1dE9 AD mice. The increase in BDNF-positive cells and decrease in activated microglia might underpin the beneficial effect.

Hypoxia-inducible factor-1alpha: molecular-targeted therapy for hepatocellular carcinoma.

Hypoxia-inducible factor (HIF)-1α is over-expressed in hepatocellular carcinoma (HCC) and degraded by ubiquitin-proteasome pathways under normoxic conditions. Hepatocyte hypoxia enhances proliferation, angiogenesis, metastasis, chemoresistance, and radioresistance of HCC. The importance role of HIF-1α expression in HCC may improve the prognostic and therapeutic technique. This article reviews the HIF-1α expression and its gene during the rat HCC development, the level of HIF-1α expression in HCC patients, and the effect of silencing HIF-1α gene by miRNA on inhibition of HepG2 cell proliferation.

[A chromosomal integration system for development of polyvalent vaccine strains].

We have developed a potential bivalent vaccine stain through the chromosomal integration system in which the gene that codes for the B subunit of cholera toxin was integrated into the chromosome of an attenuated strain of Salmonella typhimurium. This system involves two steps: construction of a hisOG deletion mutation into the chromosome of S. typhimurium strain SL3261: and replacement of the hisOG deletion by the complete hisOG region and the segment of heterologous DNA which codes for the B subunit of cholera toxin. The recombinant strain SL3261 (named TT201) was obtained. Southern hybrid confirmed that the CTB gene was integrated into the chromosome of TT201. Western blot analysis demonstrated that the CTB gene was expressed stably in TT201. When administered orally to mice, the recombinant strain elicited a serum antibody response to CTB. The strain TT201 is a potentially bivalent vaccine candidate.

[Study on zinc resistance to the immunosuppression caused by cyclophosphamidum in the mouse].

Swiss mice were used to study the effect of zinc in antagonizing the suppressive action of Cy on the immune function and thymus structure. Mice (weighing 18-22 g) receiving daily dose of 0.2 mg zinc for 25 days did not show any change in peripheral WBC count, production of IgM antibody and thymus weight, but did display marked resistance to the reduction of WBC count and number of mature T lymphocyte, suppressed production of IgM antibody and decreased thymus weight due to administration of Cy.

[Effect of zinc on immune function in normal mice].

Swiss mice were used for studying the effects of zinc on the immune function and on the structure of thymus. The experimental results showed that: 1) When mice were administered with a dose of zinc 0.2mg/d of mouse for 15 d, the number of mature T lymphocyte in peripheral blood increased significantly, the transformation of lymphocytes and DTH response were both markedly enhanced; phagocytosis of peritoneal macrophage was suppressed, but the production of IgM and the weight of thymus were not affected. 2) The shape of lymphocyte nucleus of the thymus cortex became irregular by administration of zinc. So, a certain immunity promotive dose of zinc may impose a latent injury to the lymphocyte of thymus.

[Immunoregulatory role of thyroid hormone].

The immunoregulatory role of thyroid hormone was investigated in mice weighing 20-30 g. The experimental results showed that the antibody production, transformation of lymphocytes, DTH response (delayed-type hypersensitivity response) and phagocytosis of the peritoneal macrophages were all markedly enhanced when the animals were fed with a daily dose of 2 mg thyroid hormone for 14-18 days. The number of mature T cells in peripheral blood was also increased significantly by the thyroid hormone. When mice were administered with antithyroid drug (methylthiouracil), the antibody production, the transformation of lymphocytes and DTH response were inhibited. Thyroidectomy could also suppress the antibody production. The results suggest that thyroid hormone is involved in the regulation of immune function.