With the continuous advent of new types of electron microscopes and the continuous improvement and development of electron microscope technique , biological electron microscope tech-nique has become one of the important experimental means in the fields of biology , molecular biology, basic medicine and clinical medicine , Department of Pathology of College of Basic Medical Sciences in Jilin University has reviewed the teaching feature and teaching status of biological electron microscope technique based on the training objectives of postgraduate education. By finding the problems and analyz-ing them, they have added the teaching link of review discussion in teaching process and elaborated on its implementation process and teaching results , aiming to provide practical basis for application of review discussion for postgraduate teaching.

Objective To investigate the effect of Salvia miltiorrhiza injection on proliferation , apoptosis and expression of hydroxyproline in human embryonic lung fibroblast cells. Methods TGF-β1 was administered to induce the proliferation in human embryonic lung fibroblast cells; the effect of Salvia miltiorrhiza injection on the number of human embryonic lung fibroblast cells was detected by MTT method; ki67 expression by immunocytochemical method;cell apoptosis by flow cytometry and the expression of hydroxyproline by colorimetry method. Results TGF-β1(0, 2.5, 5, 10, 20, 40 μg/L)could up-regulate cell number of human embryonic lung fibroblast cells in a dose-dependent manner , while the OD value of human embryo lung fibroblasts cells declined pretreated with Salvia miltiorrhiza injection in a dose-dependent manner and Salvia miltiorrhiza injection could induce the apoptosis and down regulated hydroxyproline expression in human embryo lung fibroblasts cells. The results of flow cytometry indicated that cell apoptosis increased after treated with Salvia miltiorrhiza injection when compared with TGF-β1 group (P < 0.05). Meanwhile, Salvia miltiorrhiza injection could down regulate the expression of hydroxyproline (P < 0.01). Conclusions Salvia miltiorrhiza injection can target human embryonic lung fibroblast cells , play a potent role in the airway remodeling through the promotion of its apoptosis and down regulate the expression of hydroxyproline.

BACKGROUND:As a new kind of adult stem cells, adipose-derived stem cells get more and more attention, because of rich source, drawing materials easily and powerful proliferation. OBJECTIVE:To isolate and culture adipose-derived stem cells from the epididymal adipose tissue in mice, and to identify their biological characteristics. METHODS:Adipose tissue was obtained from epididymis in mice by aseptical y cutting. The tissue was digested using col agenase. Adipose-derived stem cells were separated and purified by using one digestion, multiple col ection method and differential adhesion method. The morphology of adipose-derived stem cells was observed using inverted microscopy and transmission electron microscopy. Growth curve of adipose-derived stem cells was drawn. Immunophenotype of adipose-derived stem cells was identified by flow cytometry. Adipose-derived stem cells were induced to differentiate into adipocytes and osteocytes using cellinductors. Compatibility of adipose-derived stem cells and col agen scaffold material was observed using scanning electron microscope. RESULTS AND CONCLUSION:Adipose-derived stem cells exhibited long spindle-like or fibroblast-like appearance, grew intensively and arranged in scrol and fascicular shape. In vitro, adipose-derived stem cells could be passaged to passage 9 under the inverted microscope. Under the transmission electron microscope, adipose-derived stem cells showed abundant microvil i on the cellsurface. The nuclei were big in size. Some organel es were seen in cytoplasma, such as mitochondria and rough endoplasmic reticulum. Adipose-derived stem cells expressed CD44 and CD29, did not express CD34. After inducing by inductor, many smal lipid droplets were seen in the cytoplasm of adipose-derived stem cells. The smal lipid droplets were dyed red with oil red O. After induction of osteogenic inductor, the boundary line among adipose-derived stem cells was not clear and the structure of cells was fuzzy in the growth-intensive areas. There were many strong refractive granular material deposits at that field after dyeing with alizarin red. Scanning electron microscope revealed that adipose-derived stem cells were spread on the col agen scaffold. Results suggested that adipose-derived stem cells isolated by this method could amplify in vitro and stably subcultured. Under a certain inducing condition, adipose-derived stem cells could differentiate into osteoblasts and adipocytes, which showed a good compatibility with col agen scaffold.

In order to help students understand and master the ultrastructure of the tissue and cells and provide teachers with rich picture resources and teaching materials, Department of Pathology of Jilin University collected and organized pictures of cell and tissue ultrastructure, and then selected typical ultra-structure pictures to carry on the classified annotation in accordance with organ systems to build the ultra structure electronic picture library. The electronic picture library can be used in biological electron micro-scope technique course, online courses establishment and students' autonomous learning.

Objective To investigate the related mechanism of Slug inhibiting the proliferation of cervical cancer cell through CDH3/β-catenin/C-myc. Methods SiHa cells with stable Slug expression were screened. The expression of CDH3 in Slug-overexpressed SiHa cell was detected by RNA-sequence, Real-time PCR, Western blot and immunocytochemistry. The expression of CDH3 in SiHa and HeLa cells were detected by Western blot and immunocytochemistry. The protein level of CDH3 was up-regulated in HeLa cells or rescued in SiHa-Slug cells by transient transfection of CDH3 expression vector. The protein levels of β-catenin and C-myc were detected by Western blot, the cell growth was detected by cell counting and CCK-8 assays. Luciferase reporter assay and chromatin immunoprecipitation assay (ChIP) were performed to detect the effect of Slug on regulating the promoter region of CDH3. Results SiHa cell line with stable Slug expression was successfully constructed. Slug overexpression inhibited CDH3 expression in SiHa cells. CDH3 promoted cell proliferation and up-regulated the protein level of β-catenin and C-myc in HeLa and SiHa-Slug cells. Slug could recognize and bind to the E-boxes in the CDH3 promoter region and inhibited the transcription of CDH3 in SiHa cells. Conclusion Slug could inhibit the expression of β-catenin and C-myc by inhibiting CDH3 transcription in SiHa cells, and then attenuate the growth of SiHa cells.

Objective:To optimize the extraction technology and evaluate antioxidant activity of total flavonoids from Hedyotis dif-fusa. Methods:The content of kaempferol was detected by an HPLC method,and spectrophotometry was used to detect the content of total flavonoids. Results: The optimum extraction conditions were investigated by orthogonal design with the extraction quantities of kaempferol and total flavonoids as the evaluation indices. The antioxidant activities of the extracts were detected based on the clearance of hydroxyl radical and DPPH·model. The optimum extraction conditions were as follows:10-fold amount of 80% methanol and with ultrasonic extraction for 15 min. The clearance rate of 1ml methanol extract for scavenging DPPH· and ·OH was 73.89% and 91.27%,respectively.Conclusion:The optimum technology is stable and feasible for the extraction of Hedyotis diffuse. The extract shows good in vitro antioxidant properties, which provides powerful reference for the future development of relevant antioxidant prod-ucts.

ObjectiveTo systematically review the efficacy and safety of exosomes derived from bone marrow mesenchymal stem cells (BMSC) on animal models of spinal cord injury. MethodsAnimal studies about BMSC-derived exosomes for spinal cord injury were retrieved from databases of PubMed, Cochrane Library, Web of Science, CNKI and Wangfang Data, from establishment to October, 2021. Two researchers independently screened and extracted the data, and evaluated the risk of bias. The studies were qualitatively analyzed. ResultsA total of 21 studies were included, involving 1 342 animals. Male or female Sprague-Dawley rats were selected for 18 studies, and the body mass of the rats was (200±50) g in 19 studies. The injury nodes focused on T_9-11 spinal cord, with various methods. The types, medication time, frequency, concentration and dose of the exosomes were heterogeneous. ConclusionsThe BMSC-derived exosomes can improve the motor function after spinal cord injury, reduce the damage of spinal cord, resist apoptosis and inflammation, reduce the permeability of blood-spinal cord barrier, and promote the growth of axons and blood vessels. More high-quality studies are needed for further verification.

Objective: To investigate the expression of marker proteins in vocal cord leukoplakia, and to find markers for the early stage of diagnosis and prognosis of precancerous lesions. Methods: The study included 119 cases, 68 cases of vocal cord leukoplakia (22 cases with epithelial simple hyperplasia, 46 cases with epithelial dysplasia), and 51 cases of vocal cords benign lesions(31 cases of vocal cord polyps, 20 cases of Reinke′s edema). The expression of p53, Ki-67, p21, Survivin, p16, p27, PTEN, c-Myc and vascular endothelial growth factor (VEGF) in vocal cords leukoplakia were detected, vocal cord benign lesions (vocal cord polyps and Reinke′s edema) acted as controls, comparing the expression differences of different pathological tissue. Data was analyzed by SPSS 22.0 software. Results: The expression of p53, p16, Ki-67, VEGF in vocal cord benign lesions and vocal cords leukoplakia with epithelial simple hyperplasia did not show significant differences. There was a grading increase in the positive expression of p53, Ki-67 in the vocal cord leukoplakia with epithelial dysplasia contrasting to those in vocal cord benign lesions and vocal cords leukoplakia with epithelial simple hyperplasia (p53: χ²=13.340, P=0.002, Ki-67: χ²=53.386, P=0.000). The expression of p27, PTEN, c-Myc in vocal cord benign lesions and vocal cords leukoplakia with epithelial dysplasia did not show significant differences. There was a grading increase in the positive expression of p21 Survivin in vocal cords leukoplakia with epithelial dysplasia contrasting to those in vocal cord benign lesions (P<0.05). Expression of Survivin in vocal cords leukoplakia with mild-moderate epithelial dysplasia showed a significant increase than those in vocal cord benign lesions (P<0.05). The positive expression grade of p21 showed a rising trend (P=0.073) between the different grades of dysplasia. Conclusion The positive expression grade of p53, Ki-67, p21 Survivin showed an increase in vocal cords leukoplakia with epithelial dysplasia contrasting to those in vocal cord benign lesions, which might be an implication for evaluating the diagnosis and prognosis of precancerous lesions. Expression of p21 was correlated to the degrees of dysplasia and expression of Survivin showed a significant difference in early stage of epithelial dysplasia contrasting to benign lesions.

Objective: Juvenile on-set recurrent respiratory papillomatosis (JORRP) can be an aggressive and potentially life-threatening disease. To emphasize the importance of complication prevention in local excision surgery, the single-center experience of RRP treatment in a group of patients with relatively short recurrence-free-interval (>4 times/yrs) was reported. Methods: Data of 49 patients underwent papilloma resection during 2002-2013 were retrospectively analyzed. Combined techniques of laser microsurgery and microdebriders were used to remove papillomas, prevent the formation of scar tissue and improve respiratory distress. Forty-nine cases were followed up for 99.0[74.5; 121.0]months, ranging from 39 to 185 months. The age of disease onset was 2[1; 3] years old. Most of the patients had difficulty in breathing(42 of 49 cases). Seven patients had hoarseness.53.0% patients had subglottic or trachea papillomatosis spreading. The rate of complications, synechia formation, tracheal stenosis, needing for tracheostomy, mortality was recorded. Patients were followed up for at least three years. The characteristics of the patients, improvement in symptoms, recurrence-free interval, and the rate of tracheal extubation were evaluated. Wilcoxon signed ranks test and chi-square test were used for data comparation. Results: Long-term relieve in disease(≤2 surgeries were needed/year) were achieved in 29(59.1%) subjects, in which 8(16.3%) of the subjects did not need surgical treatment for at least 3 years. Long-term relieve had been achieved in 25 of subjects who did not accept tracheostomy. Seven subjects did not need surgical treatment for at least 3 years. The rate of successful tracheal extubation was 53.8% in 13 patients who had undergone tracheostomy. Three cases dead of intrapulmonary spread. Conclusions Combined techniques of laser microsurgery and microdebriders are effective in improvement in both respiratory distress and voice quality. The combined technique are helpful to avoid inevitable long-term stenotic complications, ultimately affecting the quality of life.