Objective:To study the ultrastructural changes of the junctional epithelial after dental preparation. Methods: Junctional epithelium (JE) of left premaxillary teeth was detached by diamond bur in 8 rabbits (group 1) and by surgical knife in another 8 rabbits (group 2). JE of the right teeth without treatment was used as the controls. 4 animals in each group were sacrificed 1 and 2 weeks after operation respectively.Samples were prepared for transmission electron microscope observation.Results: In the two tested groups reattachment was observed 2 weeks after operation,degeneration of mitochondria in JE cells and destruction of basement membrane were observed. No significant difference was observed between group 1 and group 2.Conclusion: Dental preparation may result in ultrastructural changes of JE cells.

Objective: To evaluate the therapeutic effect of Lishi-Jianpi-Quyu decoction combined with conventional western medicine for the oral lichen planus (OLP). Methods: A total of 80 OLP patients who met the inclusion criteria were divided into two groups by random number table method, 40 in each group. The control group was treated with conventional medicine, while the research group were treated with Lishi-Jianpi-Quyu decoction on the basis of control group. Two groups were treated for 4 weeks. The severity of oral mucosal lesions was assessed by REU scale, and oral discomfort was assessed by VAS scale. The serum tumor necrosis factor-α (TNF-α), IL-6 and IL-8 levels were detected by ELISA, peripheral blood CD3⁺, CD4⁺ levels were detected by flow cytometry, and the ratio of CD4⁺/CD8⁺ was calculatedand the clinical efficacy was evaluated. Results: The effective rate of research group was 82.5% (33/40), while the control group was 62.50% (25/40), and the difference between two groups was statistically significant (χ²=9.031, P=0.003). After treatment, TNF-α (1.19 ± 0.28 μg/L vs. 3.59 ± 0.72 μg/L, t=19.807), IL-6 (11.14 ± 2.04 ng/L vs. 16.34 ± 3.20 ng/L, t=8.666), IL-8 (1.15 ± 0.61 ng/L vs. 2.04 ± 0.76 ng/L, t=5.776) of research group were significantly lower than the control group (P<0.01). The serum CD3⁺ (64.02% ± 5.16% vs. 58.07% ± 4.85%, t=5.314), CD4⁺ (41.26% ± 3.61% vs. 37.15% ± 3.04%, t=5.508), CD4⁺/CD8⁺ (1.60 ± 0.43 vs. 1.31 ± 0.40, t=3.668) of research group were significantly higher than those of the control group (P<0.01). After treatment, the REU score and VAS score in the research group were significantly lower than that in the control group (t=10.980, 11.146, P<0.01). Conclusions The Lishi-Jianpi-Quyu decoction combined with conventional western medicine can improve the clinical symptoms of OLP, reduce the level of serum inflammatory cytokines, and enhance the body's immunity and improve the curative effect.

PURPOSE: Cancer stem cells have recently been thought to be closely related to tumor development and reoccurrence. It may be a promising way to cure malignant glioma by using glioma stem cell-targeted dendritic cells as a tumor vaccine. In this study, we explored whether pulsing dendritic cells with antigens of glioma stem cells was a potent way to induce specific cytotoxic T lymphocytes and anti-tumor immunity. MATERIALS AND METHODS: Cancer stem cells were cultured from glioma cell line U251. Lysate of glioma stem cells was obtained by the repeated freezing and thawing method. Dendritic cells (DCs) were induced and cultured from the murine bone marrow cells, the biological characteristics were detected by electron microscope and flow cytometry. The DC vaccine was obtained by mixing DCs with lysate of glioma stem cells. The DC vaccine was charactirizated through the mixed lymphocyte responses and cell killing experiment in vitro. Level of interferon-gamma (IFN-gamma) in the supernatant was checked by ELISA. RESULTS: After stimulation of lysate of glioma stem cell, expression of surface molecules of DC was up-regulated, including CD80, CD86, CD11C and MHC-II. DCs pulsed with lysate of glioma stem cells were more effective than the control group in stimulating original glioma cells-specific cytotoxic T lymphocytes responses, killing glioma cells and boosting the secretion of IFN-gamma in vitro. CONCLUSION: The results demonstrated DCs loaded with antigens derived from glioma stem cells can effectively stimulate naive T cells to form specific cytotoxic T cells, kill glioma cells cultured in vitro.
Animals ; Antigens, Neoplasm/immunology ; Apoptosis ; Brain Neoplasms/*therapy ; Cancer Vaccines/*therapeutic use ; Cell Line, Tumor ; Cell Proliferation ; Dendritic Cells/*cytology ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Glioma/*therapy ; Humans ; Interferon-gamma/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Neoplastic Stem Cells/*cytology ; T-Lymphocytes, Cytotoxic/immunology

PURPOSE: Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) are an inhibitor of receptor tyrosine kinases (RTKs) that was discovered in recent years, and many studies showed that LRIG1 is a tumor suppressor gene and may be related to tumor drug resistance. In this study, we explored whether LRIG1 protein expression can improve the chemosensitivity of glioma cells and what was its mechanism. MATERIALS AND METHODS: We collected 93 cases of glioma tissues and detected the expression of LRIG1 and BCL-2. We constructed a multidrug resistance cell line U251/multidrug resistance (MDR) and examined the change of LRIG1 and BCL-2 at mRNA and protein expression levels. LRIG1 expression was upregulated in U251/MDR cells and we detected the change of multidrug resistance. Meanwhile, we changed the expression of LRIG1 and BCL-2 and explored the relationship between LRIG1 and BCL-2. Finally, we also explored the relationship between LRIG1 and RTKs. RESULTS: LRIG1 was negatively correlated with BCL-2 expression in glioma tissue and U251/MDR cells, and upregulation of LRIG1 can enhance chemosensitivity and inhibit BCL-2 expression. Furthermore, LRIG1 was negatively correlated with RTKs in U251/MDR cells. CONCLUSION: These results demonstrated that LRIG1 can improve chemosensitivity by modulating BCL-2 expression and RTK signaling in glioma cells.
Astrocytoma/drug therapy/genetics/metabolism ; Cell Line, Tumor ; Drug Resistance, Neoplasm/genetics/*physiology ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Glioma/drug therapy/*metabolism ; Humans ; Membrane Glycoproteins/metabolism/*physiology ; Proto-Oncogene Proteins c-bcl-2/*metabolism ; RNA, Messenger/metabolism ; Receptor Protein-Tyrosine Kinases/metabolism

OBJECTIVE：To establish the rapid field identification method of Cryptotympana pustulata ecdysis. METHODS ： 3D depth of field synthesis technology was used to identify 50 batches of C. pustulata ecdysis and its adulterants from the length of beak ，size and protrusion degree of upper labial base ，the protrusion degree of the lower labial base ecdysis and the color of its upper transverse groove ，the number and shape of main and lateral spines on the foot ，significance of abdominal valves ，the number of webs ，the number and shape of side plates ，the number of tergum rings ，terminaliae，etc. RESULTS ：Among 50 batches of samples ，S1-S5，S26-S30，S36-S50 were C. pustulata ecdysis；S21-S25 was adulterants of C. pustulata ecdysis after weight gain ；S31-S35 was adulterants of C. pustulata ecdysis after extraction ；S6-S20 were ecdysis from Tibicen flammatus ，C. flammatta，Lyristes pekinensis ，all of which were adulterants. The main distinguishing feature of C. pustulata ecdysis and its adulterants was that abdomen and ventral surface of C. pustulata ecdysis were triangular ，and the abdomen and ventral surface of other species was nearly parallel ；the valve of C. flammatta ecdysis was obvious ，but those of other varieties were not obvious ；the lateral appearance of terminaliae of C. flammatta ecdysis was sharper than those of other species ；there was an acute angle between the front foot accessory thorns and the end thorns of the T. flammatus ecdysis，and an obtuse angle between the front foot accessory thorns and the end thorns of the L. pekinensis ecdysis. CONCLUSIONS ：The method is simple ，reliable and suitable for rapid field identification of C. pustulata ecdysis and its adulterants.

ObjectiveTo clarify the accumulation and distribution characteristics of dry matter and mineral elements in Artemisia argyi var. argyi cv. Qiai， and to provide technical support for the high yield of and efficient utilization of nutrients in this medicinal species. MethodTwo cultivars of this species， Qiqing 1 and Qihuang 1 were selected， and the composition of dry matter in different organs， the content， accumulation， and distribution of mineral elements in each organ of the two cultivars， and the dynamic changes of volatile oil content and index components eucalyptol and borneol in leaves of the two cultivars were monitored at different growth stages. ResultThe period from February to March marked the early growth stage of Qiai， and the dry matter was mainly distributed in the leaves. It accelerated the growth in April， and the period from April to mid-June witnessed the vigorous vegetative growth of Qiai， during which the dry matter was mainly found in the stems and leaves. It began the reproductive growth from late June and the dry matter was mainly distributed in the stems. In the flowering stage in August， no dry matter accumulation occurred. As for the volatile oil， the content was high （> 1.10%） at the vigorous vegetative growth stage and peaked on June 14 （1.33% in Qiqing 1， and 1.23% in Qihuang 1）. The relative mass fraction of eucalyptol was the maximum at the vegetative growth stage （8.67% in Qiqing 1， and 13.07% in Qihuang 1）. The relative mass fraction of borneol peaked at the early growth stage （2.63% in Qiqing 1， and 5.94% in Qihuang 1）. The content of nitrogen， phosphorus， potassium， and zinc in leaves was in significantly positive correlation with the content of volatile oil and the relative content of eucalyptol and borneol. The content of macroelements nitrogen， phosphorus， potassium， and calcium and trace elements iron and zinc peaked at the early growth stage， and the content was the highest in stem and leaf. The content of macroelement magnesium and trace elements manganese and copper was the highest at vegetative growth stage when the content of other elements decreased and the nutrients were gradually transferred to the buds， flowers and other organs. In the whole growth period， the distribution of potassium， calcium， and zinc was in the order of leaf > stem > root， and the distribution of nitrogen， phosphorus， copper， magnesium， and manganese followed the order of leaf > root > stem. The distribution of iron was in the order of root > leaf > stem. There was a significantly positive correlation between the total amount of dry matter and the absorption of nutrients in 'Qiai'. The absorption of macroelements by Qiai was in the order of potassium > nitrogen > calcium > phosphorus > magnesium， and the ratio of absorbed elements was about 2.66∶2.51∶0.6∶0.11∶0.04. The absorption of trace elements followed the order of manganese > iron > zinc > copper， and the ratio of absorbed elements was about 0.25∶0.17∶0.05∶0.04. In terms of the production of medicinal materials， 'Qiai' needed about 4.11 kg potassium， 3.58 kg nitrogen， 0.91 kg phosphorus， 0.18 kg calcium， 0.06 kg magnesium， about 6.64 g manganese， 2.56 g iron， 1.30 g zinc， and 0.92 g copper to produce 100 kg medicinal materials. ConclusionEnough organic fertilizer and phosphorus and potassium fertilizers should be applied as base fertilizers for Qiai. The vegetative growth stage （April-June） marks the high accumulation of dry matter and large demand of nutrients， during which topdressing should be conducted timely and early， especially nitrogen fertilizer， and appropriate amount of micro-element fertilizer should be added. Qiai needs a large amount of calcium and magnesium fertilizers from the mid-vegetative growth stage， and they should be applied in time in the late stage to ensure the vegetative growth of the plants for seeds and the quality of the medicinal material of Qiai.

Objective : To investigate the effect of hypoxic preconditioning (HPC) on mitochondrial energy metabolism in mouse hippocampal HT22 cells and its possible mechanism. Methods : In this paper, mouse hippocampal nerve cells HT22 were divided into control group, hypoxia group, HPC group, and the levels of adenosine triphosphate (ATP) and reactive oxygen species (ROS) in each group were measured for observing the effect of HPC on cell mitochondrial metabolism. Western blot was used to detect the expression of target of rapamycin ( mTOR), phosphorylated mTOR protein and autophagy substrate P62 protein; cellular immunofluorescence was used to detect phosphorylated mTOR, and LysoTrackerTM probe was used to detect lysosomes. Results : Compared with the control group, the ATP level was significantly decreased and the ROS level was increased in the hypoxia group. Exposed to HPC, the ATP level was increased and the ROS level was decreased. Compared with the control group, the expression of phosphorylated mTOR was down⁃regulated and the expression of autophagy substrate P62 was down⁃regulated in the HPC group. Conclusion HPC may affect the energy metabolism of HT22 cells through the mTOR/autophagy signaling pathway, thereby exerting a protective effect on the HT22 cells.