ObjectiveTo investigate the molecular mechanism by which metastasis-associated protein 3 (MTA3) participates in glioma resistance through reactive oxygen species. Methods Protein expression in glioma stem cells (GSCs) and non-GSCs was detected using Western blotting. GSCs included U87 and SHG44 cells, while non-GSCs included U87s and SU-2 cells. After overexpressing MTA3, U87 and SHG44 cells were divided into Lv-scr and Lv-MTA3 groups. The self-renewal capacity of glioma cells was assessed through a neurosphere formation assay. Cell survival fractions were examined following exposure to 0, 2, 4, 6, 8, and 10 Gy X-ray irradiation under normoxic or hypoxic conditions. Apoptosis and reactive oxygen species expression were analyzed using flow cytometry. Immunofluorescence staining was performed to detect the stem cell markers CD133 and nestin, as well as the differentiation markers glial fibrillary acidic protein (GFAP, for astrocytes) and neuronal class Ⅲ β-tubulin. Results In GSCs, MTA3 expression was lower in the U87s and SU-2 groups. After MTA3 overexpression, Lv-MTA3 expression was higher in U87s and SU-2 compared to the Lv-scr group. Under normoxic or hypoxic conditions, U87 and SU-2 showed greater radioresistance compared to glioma cell lines U87 and SHG44. Compared to non-GSCs, basal reactive oxygen species formation was reduced in GSCs, while reactive oxygen species generation was increased in non-GSCs. Following exposure to different doses of X-rays under normoxic or hypoxic conditions, GSCs with MTA3 overexpression exhibited greater radiosensitivity than those with stable integration. Additionally, MTA3 overexpression slightly increased the oxygen enhancement ratio (OER) in GSCs. MTA3 overexpression reduced the immunoreactivity of CD133 and nestin in both stem cell lines, and increased immunofluorescence staining of GFAP and neuronal class Ⅲ β-tubulin, with statistically significant differences (P<0.05). Conclusions MTA3 is downregulated in GSCs. Overexpression of MTA3 reduces the radioresistance and stemness of GSCs both in vitro and in vivo. MTA3 plays a crucial role in regulating the radiosensitivity and stemness of GSCs through reactive oxygen species.
Objective To investigate the value of thyroid stimulating hormone (TSH) inhibition therapy in postoperative patients with differentiated thyroid carcinoma. Methods One hundred and six patients with differentiated thyroid carcinoma after total or near-total thyroidectomy were divided into two groups: TSH inhibition therapy group and thyroid hormone replacement therapy group. The non-recurrence and (or) non-metastasis rate in two groups was analyzed retrospectively by Wilcoxon statistical analysis. Results The non-recurrence and (or) non-metastasis rate of 3-， 5- and 10-year in TSH inhibition therapy group were 98.31%， 92.41% and 75.45% respectively， and were higher than those (93.57%， 84.18% and 52.06% respectively) in thyroid hormone replacement therapy group （P=0.046 5）. Conclusion TSH inhibition therapy after total or near-total thyroidectomy may be an essential treatment for differentiated thyroid carcinoma patients.
Objective To investigate the effect of the 8-bromum-cyclic adenosine monophosphate (8-Br-cAMP) on the telomerase activity and changes of cell cycle in retinoblastoma （RB） cells. Methods The cultured RB cells were divided into the experimental group (8-Br-cAMP) and control group. After cultured for 24, 48 and 72 hours in vitro, the telomerase activity of RB cells was detected by polymerase chain reaction enzyme-linked immunosorbent assay (PCR-ELISA) and the changes of cell cycle were detected by flow-cytometry. Results The difference of telomerase activity was significant between the experimental groups and control group (Plt;0.01). There was a negative correlation between the A value of absorbance and the time in the experimental groups (r=-0.778 9, F=33.936, Plt;0.01). The changes of the cell cycle were that the percentages increased in G1 phase and decreased in S phases. Conclusion 8-Br-cAMP may weaken telomerase activity, affect the cell cycle, and inhibit the proliferation of RB cells. (Chin J Ocul Fundus Dis，2004，20：358-360)
From 1984 to 1993, 49 cases with varioussoft tissue defects around the knee were treated with pedicled calf myocutaneous flap, lateral sural cutaneous artery island skin flap, saphenous neurovascularskin flap and fasciocutaneous flap. The postoperation results were sucessful in 47 cases, and failure 2 cases, in one case with flap infection and theother with scar formation surrounding the knee. Both the failure cases were cured with split skin graft. The patient were followed up for an average of three and a halfyears, the knee function was almost completely regained, and the blood supply of the flaps, the elasticity and colour of the flaps were similiar to that of the normal skin, without being cumbersome. The sensation of the saphenous neurovascular flaps and the lateral suralcutaneous artery island flaps was preserved, except partial numbness was presented at the distal part of the flaps. Operative indications and selection of cases were discussed.
Objective To evaluate the potential of specific mRNA marker keratin 19(K19) to detect micrometastasis by reverse transcriptase polymerase chain reaction (RT-PCR) .Methods One hundred and ninty four regional lymph nodes harvested from 6 cases of benign diseases, 4 cases of breast carcinoma, 5 cases of gastric carcinoma and 12 cases of colorectal carcinoma patients were examined by conventional pathology and amplifying tissue specific K19 mRNA by RT-PCR separately, then the two methods were compared with each other. Results None of the 34 lymph nodes which were pathological metastasis-negative from benign diseases expressed K19 mRNA by RT-PCR, all of the 28 regional lymph nodes which were pathological metastasis-positive from malignant cases showed trains of K19 mRNA by RT-PCR. Of the 132 lymph nodes which were pathological metastasis-negative from malignant cases, 11 lymph nodes were detected with micrometastasis by genetic diagnosis.Conclusion Genetic diagnosis of lymph node micrometastasis is more sensitive than conventional pathology and has diagnostic value and merits further study.
Objective To investigate the inhibitory effects of fms-like typrosine kinase receptor sFlt-1 on retinal neovascularization (RNV).Methods Recombinant lentivirus sFlt-1（2-3）and sFlt-1（2-4）expressing the sFlt-1 (2-3) and (2-4) immunoglobulinlike regions of sFlt-1 were constructed. 96 seven-day-old C57/6J mice were randomly divided into 4 groups with 24 mice in each group. Group 1: normal control; group 2: experimental control; group 3: sFlt-1（2-3）; group 4: sFlt-1（2-4）.The mice in group 2-4 were exposed to hyperoxia with （75plusmn;2）% O2 for 5 days and then returned to normoxia with 21% O2;the mice received an intravitreal injection with 1 mu;l virus of empty vector, sFlt-1（2-3）,or sFlt-1（2-4）,respectively. Five days later, all mice underwent perfusion fluorecein angiography and retinal wholemont was made to observe the changes of retinal vessels; retinal sections were stained by hematoxylin and eosin and RNV endothelium cell nucleus were counted; vascular endothelial growth factor(VEGF) and VEGF receptor-2 (KDR/Flk-1) protein were measured by Western blot.Results Seventeen days after birth, the retinal area of fluorescein leakage and RNV, RNV nucleus which breaking through inner limiting membrane in group 3 and 4 were smaller or less than that in group 2（P＜0.01）; while VEGF protein didnprime;t changed much （P＞0.05）the expression of KDR/Flk-1 decreased significantly (P＜0.01). Conclusion sFlt-1（2-3）and sFlt-1（2-4）can inhibit the formation of oxygen-induced RNV,the former virus has a better effect.
To investigate the relationship between clinicopathological features and lymph node metastasis in the primary gastric cancer and affer the basis for deciding appropriate extent of lymph node dissection, a total of 192 patients who underwent curative gastrectomy and lymph node dissection for gastric cancer were analyzed retrospectively. Result: The total rate of lymph node metastasis was 60.4%, with 28.9% of the resected lymph nodes involved. The lymph node metastasis of C, M, A region and the whole stomach were 64.6%, 57.7%, 59.1% and 90.9% respectively. The rates of the lymph node metastasis increased successively in carcinoma of early, middle and late stages (Ｐ＜0.05), the rate of the infiltrative tumor (Borr Ⅲ，Ⅳ) being 76.5% which was significantly higher than that of the circumscribed tumor (Borr Ⅰ，Ⅱ) (43.2%)。 Relating with the tumor size ＜4cm in diameter showed lesser rate, while 4-8 cm and ＞8cm in diameter showed increasingly higher metastaticrate (Ｐ＜0.01). As a result, we should decide the appropriate extent of lymph node dissection during the operation on the basis of clinicopathological stages, type of Borrmann’s, site and maximum diameter of gastric cancer along with the state of lymph node metastasis in carcinoma of different region of the stomach.
ObjectiveTo study and analyse the correlation between biologic behavior and clinical factors in gastric cancer.MethodsClinical and pathological study of carcinoma of stomach were retrospectively made in 1034 patients. ResultsIn this series,148 of 1034 patients (14.3%) were early gastric cancer.The frequency of lymph nodes metastasis was higher in proximal gastric cancer than distal (P＜0.0001).Similar frequency can also be seen in the tumor of larger diameter (P＜0.01),deeper invasion (P＜0.0001) and poor differentiation (P=0.004).Some difference in ages and sex of patients may be found on the invasion (P=0.003),differentiation (P＜0.0001),site (P＜0.001) and frequency of lymph nodes metastasis of the tumor (P=0.01).In multifactorial multivariate linear regression analysis,the site of tumor (P=0.003),diameter of tumor (P＜0.0001),depth of tumor infiltration (P＜0.0001) and the cell differentiation showed significant association with lymph node metastasis,in which the female patient had more lymph node metastasis than male (P＜0.001).Depth of tumor infiltration was the most important factor in lymph node metastasis.Numbers of lymph nodes resected were much more in total and distal gastrectomies than that in proximal gastrectomy (P＜0.0001). ConclusionThe results of this study suggest that radical gastrectomy with lymphadenectomy is necessary even in all stages of gastric cancer.