Tetrachlorodibenzo-p-dioxin-induced cleft palate because of partial loss of cell polarity to interfere with apoptosis during early developmental stage / 中华口腔医学杂志

<p><b>OBJECTIVE</b>In this study, folic acid (FA) was tested for antiteratogenic effects on Tetrachlorodibenzo-p-dioxin (TCDD)-induced cleft palate in fetal mice.</p><p><b>METHODS</b>In the present study, pregnant mice were dosed with TCDD 24 µg/kg and with or without FA 5 mg/kg body weight on gestation day 10. Control group mice received sesame oil 50 ml/kg body weight on gestation day(GD)10. The mice were sacrificed on GD12.5, GD13.5, GD14.5, GD15.5 and GD16.5. From each pregnant mouse on GD16.5, embryos were obtained to examine under a dissecting microscope, and routine histology was performed for detection and classification of palatal clefts. The fetuses were prepared for histologic examination, scanning electron microscope and TdT-mediated X-dUTP nick and labeling (TUNEL). On GD12.5, GD13.5, GD14.5 and GD15.5. Meanwhile, real-time (RT)-PCR was employed to detect the mRNA expression levels about arylhydrocarbon receptor (AHR) and transforming growth factor (TGF)β3 in this animal model.</p><p><b>RESULTS</b>Total frequencies of clefts were 70.2% in TCDD group(group B) and 66.3% in TCDD+FA group(group C) in relation to control fetuses(group A). Filopodia disappeared completely at the medial edge epithelia surface on GD15.5 (group A), GD12.5 (group B) and GD14.5 (group C). the RT-PCR results showed that TGF -β3 expression was down-regulated on GD13.5 and GD14.5 compared to the control.</p><p><b>CONCLUSIONS</b>It is found that folic acid has no protects agaist 2.3.7.8-TCDD-indued cleft palate in the experiment. Meanwhile, TCDD repressed the TGF-β3 expression during the palatal development. Anormal apoptosis was induced by 2, 3, 7, 8-TCDD at the medial edge epithelia (MEE) during the early development stage.</p>

"Transplant human PBMC to establish a ""human-mouse""xenogeneic graft-versus-host disease model" / 中国免疫学杂志

Objective:To explore the suitable mouse strains,and establish stable human-miceX-GVHD model.Methods:This study selected the Nude Mice and NOD/SCID, and gave them sublethal dose of γ-ray irradiation whole body , and then intraperitoneal transplanted human peripheral blood mononuclear cells ( PBMC) to establish xenogeneic acute graft-versus-host disease model.By detection of human T cells in the mice′s tail venous blood,tissues,organs of the infiltration and other indicators (By flow cy-tometry and immunohistochemistry ) ,we compared the human immune cell infiltration rates in the two mouse model and recorded the survival time.Finally,we determined the appropriate strains of mice to establish X-GVHD.Optimization means were transfer ways and the appropriate amount of human PBMC ,and the best time to observe the changes of T cell phenotype and function.Results:The NOD/SCID mouse was more suitable for inducing human-mouseX-GVHD model,and there were no significant differences between intrap-eritoneal injection and intravenous injection.Transfer human PBMC more than 5 ×107 can establish human-mouseX-GVHD model.Using the optimized experimental conditions to establish the human-mouseX-GVHD model,we found the 7-11 days was the best time to observe the changes of T cell phenotype and function ,and the average survival time was(14.16±1.77)days.Conclusion:Human-miceX-GVHD model can be successfully established by intraperitoneal injection of 5×107 human PBMC into NOD/SCID, and the best time to observe the changes of T cell phenotype and function is between 7-11 days.

Establishment of palatal organ culture in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The purpose of this study was to establish a palatal organ culture method and to investigate the palatogenesis in vitro.</p><p><b>METHODS</b>20 pregnant 14-day mice were killed, embryos were separated ascetically, and palatal shelves were dissected and placed on a modified Trowell's system. All explants were cultured 24 h and 48 h respectively. Finally, all explants were embedded and stained by Hematoxylin and Eosin.</p><p><b>RESULTS</b>All explants grew healthy. After incubation for 24 h, medial edge epithelium maintained, whereas after 48 h, medial edge epithelium disappeared, bilateral mesenchymal cells contacted, palates fused.</p><p><b>CONCLUSION</b>This method provides an effective way for investigating the etiology of cleft palate in vitro.</p>

Investigation on Dexamethasone induced cleft palate in embryos of C57BL/6J mice by 1H-NMR pattern recognition of metabolites group / 生物医学工程学杂志

Metabonomics approach is a science that systematically studies the regularity of changes of metabolites and reveals the nature of metabolic activities of the lives in the dynamic process of metabolism. In this study, pregnant C57BL/6J mice were randomly divided into two groups with 15 mice in each. The ones in the experimental group were intraperitoneally injected with Dexamethasone and the others in the control group with isotonic Na chloride from 10th to 12th day of gestational period. On 17.5th day, all the mice were executed. The livers were extracted and prepared into aqueous soluble liver tissue extracts. The technology of nuclear magnetic resonance (NMR) was used to detect the endogenous small molecule metabolites. Finally, through the method of principal component analysis (PCA), changes of metabolites ingredients could be determined. The experimental results showed that there was significant difference in PCA scores plot between the two groups. Furthermore, the difference was in line with that of incidence of cleft palate in the embryos between the two groups. Therefore, metabonomics results can be used to reflect the changes of metabolites group interfered by Dexamethasone in the course of pregnancy of C57BL/6J mice and this method opens up a new way for further study of pathogenic mechanism of cleft lip with or without palate.

Comparative study on cancer cell apoptosis between gastric and intestinal-type human gastric carcinoma / 华中科技大学学报（医学）（英德文版）

Apoptosis of cancer cells between the gastric and intestinal-type human gastric carcinoma were compared in terms of the expression of oncogene MDM2 and CD68, the histological types, the infiltration depth, and lymph node metastasis. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) assay was employed to stain apoptotic cells. Histochemical method(AB-PAS) was applied to stain mucus that is neutral or acidic in nature. Immunohistochemical method (SABC) was used to detect expression of MDM2 and CD6. The results showed that the mean apoptosis index (AI) of total 48 cases was 8.60+/-2.60. AI in the 30 intestinal type cases was significantly higher than that in the 18 gastric type cases (t=4.67, P0.05). But in the gastric type cases, a significant difference existed (t=5.87, P<0.01). A significant difference in lymph node metastasis ratio was found between the two gastric carcinoma types (chi2=4.48, P<0.05). The CD68 expression ratio in the 30 intestinal type cases was much lower than that in the 18 gastric type cases (t=4.29, P<0.01). AI of 25 MDM2-positive cases was much lower than that of the 23 MDM2-negative cases (t=7.80, P<0.01). CD68 positive ratio in the 25 MDM2-negative cases was much lower than that in the 23 negative cases. The difference was statistically significant (t=10.90, P<0.01). Except for few cells scattering within the cancer nest, most CD68 positive cells infiltrated in the interstitium around the cancer tissue. In the high-AI cases, CD68-positive cells increased. And the CD68-positive cells decreased in low-AI cases (r=0.96, P<0.01). Logistic regression analysis suggested that among the control variables, only AI was a statistically significant factor in the regression model (chi2=9.64, P<0.01). We concluded that (1) the spontaneous apoptosis index in gastric-type cases of gastric carcinoma was significantly lower than that in intestinal type cases; (2) AI in the two types was influenced by the expression of MDM2 and lymph node metastasis, but no visible connection was found between AI and the infiltration depth or histological types; (3) in the intestinal type cases, AI and the CD68-positive cells increased in MDM2-negative cases.

Comparative Study on Cancer Cell Apoptosis between Gastric and Intestinal-type Human Gastric Carcinoma / 华中科技大学学报（医学）（英德文版）

Apoptosis of cancer cells between the gastric and intestinal-type human gastric carcinoma were compared in terms of the expression of oncogene MDM2 and CD68, the histological types, the infiltration depth, and lymph node metastasis. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) assay was employed to stain apoptotic cells.Histochemical method(AB-PAS) was applied to stain mucus that is neutral or acidic in nature. Immunohistochemical method (SABC) was used to detect expression of MDM2 and CD6. The results showed that the mean apoptosis index (AI) of total 48 cases was 8.60±2.60. AI in the 30 intestinal type cases was significantly higher than that in the 18 gastric type cases (t=4.67, P＜0.01). In the 30intestinal type cases, the spontaneous apoptosis index of MDM2 negative cases was significantly higher than that of the positive cases (t=7.16, P＜0.01). And in the 18 gastric type cases, the same result was found. (t=11.39, P＜0.01). The MDM2 positive ratio in gastric type cases was higher than that in intestinal type cases (x2=4.68, P＜0.05). There is no significant difference in AI between cases of lymph node metastasis and non-metastasis cases in intestinal type cases (t=0.26, P＞0.05). But in the gastric type cases, a significant difference existed (t=5.87, P＜0.01). A significant difference in lymph node metastasis ratio was found between the two gastric carcinoma types (x2=4.48, P＜0.05).The CD68 expression ratio in the 30 intestinal type cases was much lower than that in the 18 gastric type cases (t=4.29, P＜0.01). AI of 25 MDM2-positive cases was much lower than that of the 23MDM2-negative cases (t=7.80, P＜0.01). CD68 positive ratio in the 25 MDM2-negative cases was much lower than that in the 23 negative cases. The difference was statistically significant (t=10.90,P＜0.01). Except for few cells scattering within the cancer nest, most CD68 positive cells infiltrated in the interstitium around the cancer tissue. In the high-AI cases, CD68-positive cells increased. And the CD68-positive cells decreased in low-AI cases (r=0.96, P＜0.01). Logistic regression analysis suggested that among the control variables, only AI was a statistically significant factor in the regression model (x2=9.64, P＜0.01). We concluded that (1) the spontaneous apoptosis index in gastric-type cases of gastric carcinoma was significantly lower than that in intestinal type cases; (2) AI in the two types was influenced by the expression of MDM2 and lymph node metastasis, but no visible connection was found between AI and the infiltration depth or histological types; (3) in the intestinal type cases,AI and the CD68-positive cells increased in MDM2-negative cases.