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Objective:To investigate the effect and regulation of umbilical cord-derived mesenchymal stem cells (UC-MSCs) on islets function and NOD-like receptor family, pyrin domain containing 3 (NLRP3) and autophagy in type 2 diabetic mellitus (T2DM) mice.Methods:Experimental study. Twenty, 8-week-old, male C57BL/6J mice were selected and divided into a normal control group ( n=5) and a high-fat feeding modeling group ( n=15). The model of T2DM was established by high-fat feeding combined with intraperitoneal injection of low-dose streptozotocin. After successful modeling, those mice were divided into a diabetes group ( n=7) and a UC-MSCs treatment group ( n=7). The UC-MSCs treatment group was given UC-MSCs (1×10 6/0.2 ml phosphate buffer solution) by tail vein infusion once a week for a total of 4 weeks; the diabetes group was injected with the same amount of normal saline, and the normal control group was not treated. One week after the treatment, mice underwent intraperitoneal glucose tolerance tests and intraperitoneal insulin tolerance tests, and then the mice were sacrificed to obtain pancreatic tissue to detect the expressions of interleukin-1β (IL-1β) and pancreatic and duodenal homeobox 1 (PDX-1) by immunofluorescence. The bone marrow-derived macrophages were stimulated with lipopolysaccharide and adenosine triphosphate (experimental group) in vitro, then co-cultured with UC-MSCs for 24 h (treatment group). After the culture, enzyme-linked immunosorbent assay was used to detect the secretion level of IL-1β in the supernatant, and immunofluorescence staining was used to detect the expression of NLRP3 inflammasome, and related autophagy proteins. Statistical analysis was performed using unpaired one-way analysis of variance, repeated measure analysis of variance. Results:In vivo experiments showed that compared with the diabetes group, the UC-MSCs treatment group partially repaired islet structure, improved glucose tolerance and insulin sensitivity (all P<0.05), and the expression of PDX-1 increased and IL-1β decreased in islets under confocal microscopy. In vitro experiments showed that compared with the experimental group, the level of IL-1β secreted by macrophages in the treatment group was decreased [(85.9±74.6) pg/ml vs. (883.4±446.2) pg/ml, P=0.001], the expression of NLRP3 inflammasome and autophagy-related protein P62 was decreased, and the expressions of microtubule-associated protein 1 light chain 3β (LC3) and autophagy effector Beclin-1 were increased under confocal microscopy. Conclusions:UC-MSCs can reduce the level of pancreatic inflammation in T2DM mice, preserving pancreatic function. This might be associated with the ability of UC-MSCs to inhibit the activity of NLRP3 inflammasomes in macrophages and enhance autophagy levels.
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Objective:To investigate the correlation between histologic chorioamnionitis(HCA) and periventricular leukomalacia(PVL) in preterm infants less than 34 weeks old.Methods:A total of 287 preterm infants born in Qingdao Women′s and Children′s Hospital from January 2018 to December 2018, whose mothers underwent placental pathological examination and preterm infants transferred to the neonatal intensive care unit for treatment, and whose gestational age was less than 34 weeks old, were selected as the study objects. According to the results of placental pathological examination, the infants were divided into two groups: the positive HCA group(167 cases)and the negative HCA group(120 cases). The incidence of PVL was compared between the two groups. According to the results of placental pathological examination and the stage standard of HCA, the preterm infants who had been diagnosed with PVL(41 cases) were divided into three groups: the non-HCA group, the early HCA group and the middle/late HCA group.The severity of PVL, clinical data, complications were compared in each groups, and the conditions that following up to 6 months were adjusted.Results:PVL was 19.16%(32/167) in the positive HCA group and was 7.50%(9/120) in the negative HCA group.There was significant difference in the incidence of PVL between the two groups( P<0.05). Among the preterm infants with PVL, 21.95%(9/41) was in non-HCA group, 31.71%(13/41) was in the early HCA group, and 46.34%(19/41) was in the middle/late HCA group.The severity of PVL, 1 min Apgar score, white blood cell count at 24 h after birth, the incidence of bronchopulmonary dysplasia, the number of hospital stay, the use of antibiotics, the mental development index(MDI) and psychomotor development index(PDI) at the adjusted gestational age to 6 months were significant differences among the three groups( P<0.05). Moreover, the degree of HCA inflammation was positively correlated with the severity of PVL( r s=0.374, P=0.016). Conclusion:There is a correlation between HCA and PVL in premature less than 34 weeks old.With the increasing of HCA inflammation, the incidence and severity of PVL increase. With the progression of the severity of inflammation, the white blood cell count at 24 h after birth, the incidence of bronchopulmonary dysplasia, the use of antibiotics and the time of hospital stay increase, the MDI and PDI scores at the adjusted gestational age to 6 months decrease.
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Here we reported a research project based on Black-board to integrate medical curriculum .The key points of this research is application of clinical cases as teaching data and facilitate learning of knowledge following the principle of learning by doing and , input the concept of precision medicine and informatics in learning process with an individually designed framework of learning .The learning outcome is evaluated with big data tech-nology and thus creates a student-centered pathway of medical education .
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Aim To investigate the effect of artesunate on the invasion of human colon cancer Lovo cells and the possible mechanisms. Methods After Lovo cells were treated with different doses of artesunate(20,80, 160 μmol·L - 1 ), the soft agar colony formation test was adopted to observe the anchorage-independent pro-liferation of Lovo cells. Transwell assay was used to determine the effect of artesunate on the invasion abili-ty of Lovo cells. And the protein expressions of HMGB1 and MMP-2 were investigated by western blot. Results Artesunate could significantly inhibit both proliferation and invasion ability of Lovo cells in a dose-dependent manner(P < 0. 01). The experimental group treated with artesunate significantly down-regula-ted the protein expressions of HMGB1 and MMP-2 compared with control group(P < 0. 05). Conclusion Artesunate could inhibit the invasion of human colon cancer Lovo cells by down-regulating HMGB1 and MMP-2 expressions.
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Curcumin can suppress the proliferation of esophageal cancer cells by inducing cell cycle arrest and blocking Notch signaling pathway,and can inhibit the invasiveness of esophageal cancer cells by inhibiting the nuclear factor-κB (NF-κB) signaling pathway and suppressing the formation of tumor blood vessels and inhibiting matrix metalloproteinase (MMP).Curcumin can also regulate apoptosis of esophageal cancer cells by changing the expression and localization of nucleophosmin.
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Aim To investigate the promoting apoptosis effect of artesunate( ART) on human colon cancer Lovo cells and its mechanisms. Methods MTT assay was performed to determine the anti-proliferative effect of artesunate. Flow cytometry assay and electron micros-copy( EM) were used to evaluate the apoptotic effect of artesunate. Luciferase reporter assay was introduced to measure the activation of Wnt/β-catenin pathway. Western blot was used to detect the pathway-related protein levels of β-catenin, GSK-3β,c-Myc and apop-tosis-related protein level of casepase-3 . Results Compared with the control group, the inhibitory rate of cell proliferation at 72 h and 320 μmol·L-1 ART was (78. 99 ± 1. 95 )% ( F =898. 301, P =0. 000 ); the cell apoptotic rate at 24 h and 160 μmol · L-1 ART was(19. 00 ± 0. 05)% and morphological signs of cell apoptosis were found by EM;the transcriptional activi-ty of TCF4/LEF at 24 h and 160 μmol·L-1 ART was (0. 36 ± 0. 30)%(F =470. 954,P <0. 01); the ex-pressions of caspase-3 and GSK-3β were significantly increased, whileβ-catenin and c-Myc were significant-ly decreased when treated with different concentrations of ART for 48 h ( P <0. 01 ) . Conclusion ART may significantly inhibit proliferation and promote apoptosis of Lovo cells probably by inactivating Wnt/β-catenin pathway.
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Curcumin can induce cell apoptosis in human chondrosarcoma cells through extrinsic death receptor pathway,inhibit proliferation of lung cancer cells by inducing cell cycle arrest,and can suppress proliferation and induce apoptosis in cholangiocarcinoma cells through blocking multiple signaling pathways,suppress the formation of tumor blood vessels by reducing the expression of vascular endothelial growth factor in Ehrlich ascites cancer cells,inhibit breast cancer cell motility and invasiveness by regulating the expression of adhesion molecules and increase the sensitivity of chemotherapy drugs to cancer cells by regulating the expression of multidrug resistance related genes.Curcumin has explored new approaches for the treatment of tumor.
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Purpose To study the expression and interaction of iNOS and COX-2 in colorectal adenocarcinoma, as well as their relationship with the biological behaviors of colorectal adenocarcinoma.Methods Intestinal biopsy specimens of colorectal adenocarcinoma were collected in the 78 cases and 33 normal intestinal mucosal tissues.The expression of iNOS and COX-2 proteins was detected by immunohistochemical staining (SP method).Results The positive rates of iNOS and COX-2 protein was significantly higher in normal intestinal mucosa than that in colorectal adenocarcinoma (P<0.05).The expression of iNOS and COX-2 protien had significant relation with lymph node metastasis (P<0.05).The positive expression of iNOS and COX-2 in intestinal adenocarcinoma was related with TNM stage:the positive expression in patients with Ⅲ+Ⅳ stage was higher than that with Ⅰ+Ⅱstage (P<0.05). The expression of iNOS was closely correlated with COX-2 (P<0.05).Conclusions The overexpression of COX-2 and iNOS participates in the pathogenesis of colorectal carcinoma and is associated with lymph node metastasis and TNM stage of colorectal adenocarcinoma.The expression of iNOS is correlated with COX-2 in the cancer.
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<p><b>OBJECTIVE</b>To explore the molecular events and mechanism in the carcinogenesis of esophageal epithelium in the high incidence area of esophageal carcinoma.</p><p><b>METHODS</b>Epithelial cells collected from the high incidence area of esophageal carcinoma were used to detect DNA content and ploidy by propidium iodide(PI) stain. The expressions of p53, p16 and cyclin D1 were stained by indirect immunofluorescence of fluorescein isothiocyanate(FTTC), which were detected by flow cytometry (FCM).</p><p><b>RESULTS</b>During the process of carcinogenesis, DNA content increased significantly. The diploid cells decreased while heteroploid cells increased sharply, with a heteroploidy rate of 84.2%. At the same time, the p53 protein accumulated and p16 was deleted. The positive rates of p53 and oncogene cyclin D1 were both 100%(5/5, 6/6) in the cancer group.</p><p><b>CONCLUSION</b>In the early carcinogenesis of esophageal epithelium, DNA content and heteroploidy rates increase with tumor suppressor gene p16 deletion and p53 protein accumulation while oncogene cyclin D1 is overexpressed. Multiple molecular events have already occurred when esophageal carcinoma develops.</p>
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Humans , Cyclin D1 , Metabolism , Cyclin-Dependent Kinase Inhibitor p16 , Metabolism , DNA, Neoplasm , Metabolism , Esophageal Neoplasms , Genetics , Metabolism , Pathology , Flow Cytometry , Gene Expression , Precancerous Conditions , Genetics , Tumor Suppressor Protein p53 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of Matrine on apoptosis of fibroblasts and the expression of apoptotic modulation related protein in the hypertrophic scar.</p><p><b>METHODS</b>Hypertrophic scar was produced on the ear of 24 New Zealand white rabbits, which were employed as the model, and were randomly and equally divided into control (CC) and Matrine (M) groups (12 in each group). Matrine (50 g/L) was injected into the ear scar in M group and with normal saline in C group once every four days. At 2, 4, 6, 8 and 12 weeks after the injection, the apoptotic fibroblast count in the scar was determined by TUNEL method, and the expressions of apoptosis related modulation proteins p53, bcl-2, bax were detected by immunohistochemistry method.</p><p><b>RESULTS</b>The apoptotic fibroblast count was much larger in M group than that in C group at all test time points (P < 0.05). Furthermore, the bax expression was increased and that of p53 and bcl-2 was decreased significantly in M group. In adding, the scar became flat in M group.</p><p><b>CONCLUSION</b>Matrine might obviously enhance the fibroblast apoptosis in rabbit ear hypertrophic scar, and up-regulate the expression of apoptosis related modulation protein bax and down-regulate the expression of p53 and Bcl-2.</p>
Subject(s)
Animals , Female , Male , Rabbits , Alkaloids , Pharmacology , Apoptosis , Cicatrix, Hypertrophic , Metabolism , Pathology , Fibroblasts , Immunohistochemistry , Proto-Oncogene Proteins , Proto-Oncogene Proteins c-bcl-2 , Quinolizines , Tumor Suppressor Protein p53 , bcl-2-Associated X ProteinABSTRACT
<p><b>OBJECTIVE</b>To investigate the influence of the endothelial cells on the scar-derived fibroblasts in the hypertropic scar tissue.</p><p><b>METHODS</b>The endothelial cell from the human umbilical vein was cultured and its supernatant was then collected. Thereafter, it was mixed with the pre-cultured scar-derived fibroblasts from the hypertropic scar tissue. The fibroblasts were analyzed with a flow cytometer in a MTT-assay method to evaluate the cell proliferation and its division cycle.</p><p><b>RESULTS</b>The endothelial cells were significantly increasing the scar-derived fibroblast proliferation (P < 0.05), compared with the control. The proportion of the fibroblasts in the S-stage of cell cycle was also quite high.</p><p><b>CONCLUSION</b>The endothelial cells may secrete some active products, which may enhance the proliferation of the scar-derived fibroblasts and stimulate the fibroblasts into the S-stage of cell generation cycle.</p>
Subject(s)
Humans , Cell Cycle , Cell Division , Cell Line , Cells, Cultured , Cicatrix, Hypertrophic , Pathology , Culture Media, Conditioned , Chemistry , Pharmacology , Dose-Response Relationship, Drug , Endothelium, Vascular , Chemistry , Cell Biology , Fibroblasts , PathologyABSTRACT
<p><b>OBJECTIVE</b>To summarize and present the experiences and results of the expanded distant skin flaps used in plastic and reconstructive operations.</p><p><b>METHODS</b>102 patients who had undergone expanded distant skin flap transfer were reviewed and followed up.</p><p><b>RESULTS</b>Good results were obtained. Especially when there was no sufficient normal skin close to the lesion or defect, the distant skin flaps were employed. The expanded distant skin flap can be directly transferred as a pedicle skin flap or an island skin flap. It can be indirectly transferred with an intermediary carrier or as a free skin flap.</p><p><b>CONCLUSION</b>The expanded distant skin flaps have many advantages, indications, worthy of recommendation.</p>
Subject(s)
Humans , Skin Transplantation , Methods , Surgery, Plastic , Methods , Surgical FlapsABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of supernatants of normal skin keratinocytes(NK) and scar keratinocytes(SK) on proliferation and collagen synthesis of hypertrophic scar fibroblasts(HSFB).</p><p><b>METHODS</b>The supernatant, collected from cultured NK and SK, was added to the cultivated HSFB. The MTT-method, 3H-proline incorporation and radioimmunoassay were employed to measure the cell proliferation, collagen synthesis and secretion.</p><p><b>RESULTS</b>NK supernatant could inhibit HSFB proliferation and increase the collagen synthesis, but inhibit collagen secretion, as compared with the control group. On the contrary, SK supernatant could increase collagen synthesis and secretion, which had little effects on HSFB proliferation.</p><p><b>CONCLUSION</b>Keratinocytes derived from normal skin and hypertrophic scar show different effects on hypertrophic scar fibroblasts.</p>
Subject(s)
Humans , Cell Division , Cells, Cultured , Cicatrix, Hypertrophic , Metabolism , Pathology , Collagen , Culture Media, Conditioned , Fibroblasts , Physiology , Keratinocytes , PhysiologyABSTRACT
Objective To study the influnence of estrogen and progestin on scar formation. Methods By culturing hyperplastic scar fibroblast(HSFB), we investigated TGF ? 1 synthesis by immunohistochemical staining and image analysis. Results The detectable level of TGF ? 1 in HSFB treated with 17 ? E 2 was higher than that of the control significantly( P 0.05). Conclusion In vitro, 17 ? E 2 can stimulate TGF ? 1 synthesis in HSFB significantly.
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Objective To investigate the mechanism and effect of TMP on melanocytes.Methods MTT method, NaOH-assay and Takahashi method were employed to measure the proliferation, melanin synthesis, tyrosinase activity of melanocytes. Results TMP induced a mild effect on melanocytic proliferation ( p