Expression of circ_0006692 in Non-small Cell Lung Cancer and Its Regulatory Mechanism on Proliferation and Metastasis of Lung Cancer Cells / 肿瘤防治研究

Objective To explore the expression of circ_0006692 in NSCLC and its relation with clinicopathological characteristics and related mechanism. Methods We collected 50 pairs of NSCLC tissues and adjacent tissues. The expression of circ_0006692 was detected by qRT-PCR and its relation with clinicopathological features was analyzed. We constructed lung cancer A549 cell line with circ_0006692 overexpression and knockdown. Cell proliferation, migration and invasion were detected by MTS, colony forming, wound healing and Transwell invasion assays. qRT-PCR and Western blot were used to detect the effect of circ_0006692 expression change on EMT-related gene expression. Results The expression of circ_0006692 in NSCLC was significantly higher than that in adjacent tissues (P < 0.05). The expression level of circ_0006692 was closely related to tumor size, TNM stage and pulmonary membrane invasion (P < 0.05). circ_0006692 knockdown inhibited cell proliferation, invasion and metastasis, while its overexpression promoted cell proliferation, invasion and metastasis. circ_0006692 knockdown inhibited the expression of EMT-related genes CDH2 and MMP7 in A549 cells and promoted the expression of CDH1. Conclusion The upregulated expression of circ_0006692 in NSCLC is closely related to tumor size, TNM stage and pulmonary membrane invasion. circ_0006692 expression could regulate the proliferation, invasion, metastasis and EMT progression of lung cancer A549 cells.

A new pyrazolo4,3-i/pyriinidine derivative-. Synthesis and anti-inflammatory activity / 中国药理学通报

Aim To design and synthesizea new pyrazolo[4,3-d] pyrimidine derivative for the purpose of developing novel anti-inflammatory agents, and to revealthe possible anti-inflammatory mechanisms of thenew compound using preliminary studies. Methods The changes of the cell morphology were investigated via the microscope. The influence of title compound on the NO production of the L P S - stimulated RAW264. 7 cells was measured by Griess assay. The gene relative transcription levels of TNF-a, IL - 6 and IL-lß were evaluated by qRT-PCR. COX-2 expression was measured by qRT-PCR and western blot. NF-KB/NLRP3 inflammasome signaling pathway were investigated by Western blot. The changes of lung tissue were observed by HE staining in vivo experiments of mice. Results Preliminary mechanism researches revealed that the ti - tle compound could inhibit nitric oxide (NO) generation as well as the expression of COX-2, TNF-a, IL - 6, and IL - lβ by NF-KB/NLRP3 inflammasome signaling pathway in RAW264. 7 cells. Furthermore, it could simultaneously improve the morphology of the cells and relieve acute lung injury in mice. Conclusions The new pyrazolo [4, 3-α] pyrimidine derivative has anti-inflammatory activity through NF-KB/NLRP3 inflammasome signaling pathway.

Isolation and identification of impurities from raw material of clindamycin phosphate / 药学学报

Four impurities were isolated from raw material of clindamycin phosphate (CP), and their structures have been determined. LC-MS was used to determine the molecular weights of the impurities in the raw material of CP. Reversed-phase preparative HPLC was used to prepare them, and their chemical structures were identified by HR-MS and NMR. The four unknown impurities were determined as clindamycin-B-phosphate (1), clindamycin-2,4-diphosphate (2), 3',6'-dehydro clindamycin phosphate (3), epi-clindamycin phosphate (4). Impurity 1 has been included in BP and EP, while 2, 3 and 4 have not. The impurities 2, 3, 4 are first separated from raw material of CP.

Analysis of the clinicopathological features of hepatocellular carcinoma in elderly patients.

The peak age of onset of hepatocellular carcinoma (HCC) is continually increasing worldwide. This study aims to evaluate whether there exists any significant difference in the clinicopathological features between younger- and elderly-HCC.1082 Consecutive patients with HCC who underwent liver resection at Liver Cancer Institute, Zhongshan Hospital, Fudan University from 1995 to 1998 were studied. The patients were divided into elderly-HCC (>or=65 years of age) and younger-HCC (< 65 years of age). Important clinicopathological features of the patients and postoperative survival rates were compared between these two groups. Among 1082 patients studied, 108 were elderly-HCC and 974 were younger-HCC. The resection rate of the elderly-HCC was significantly higher than that of the younger-HCC. The 1, 3 and 5-year survival rates of the elderly-HCC were not significantly different from those of the younger-HCC. Compared with the younger-HCC, the elderly-HCC had (1) less HBsAg-positive rate; (2) more frequent anti-HCV positivity ; (3) lower proportion with AFP value >or=400 microg/dl; (4) a relatively small tumor diameter; (5) higher proportion of stage I-II patients; (6) a relatively low metastasis rate. However, there were no statistically significant differences in other clinicopathological features (including gender, symptoms, tumor number, tumor venous invasion, tumor differentiation, capsular formation, type of cirrhosis) between the two groups. There is a certain extent difference in clinicopathological features between elderly and younger-HCC patients, but the postoperative survival rate is comparable between the two groups.

Infectious bovine rhinotracheitis viral gG expression and gG-ELISA development / 生物工程学报

Taking the genome DNA of Infectious Bovine Rhinotracheitis Virus (IBRV) as the template, the gG gene was amplified with PCR and cloned into the T cloning vector pMD18-T. After being identified by restriction digestion and DNA sequencing, the insert was subcloned into the expression vector pGEX-KG. Sodium docecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot assay showed that this gene was expressed as both soluble form and inclusion body by the transformed E. coli BL21 strain (DE3). The fusion protein was purified and used as the coating antigen to develop the indirect Enzyme-Linked Immunosorbent Assay (ELISA). Comparison between this gG-ELISA and commercial IBRV gB-ELISA Kit (IDEXX) was made in the detection of 380 cow serum samples. The results demonstrated an agreement of 92%. By using this novel gG-ELISA, 1248 cow serum samples were tested and the average positive rate of IBRV antibodies for imported cows is 21.7%, while the positive rate ranged greatly from 0.0%-41.5% for Hubei local Chinese Black and White Dairy Cows.

Deletion of OPCML gene and promoter methylation in ovarian epithelial carcinoma / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the expression of OPCML gene in ovarian epithelial carcinoma and determine the relationship between mRNA expression and methylation of their promoters.</p><p><b>METHOD</b>Twenty normal ovarian tissues and 89 ovarian epithelial tumor specimens (72 malignant, 17 benign), as well as 3 ovarian carcinoma cell lines (SKOV-3, CAOV3, and 3AO), were collected for detection of OPCML gene expression by reverse transcription-polymerase chain reaction and for detection of promoter methylation by restriction enzyme cut analysis from 7. 1999 to 7. 2003.</p><p><b>RESULTS</b>Among ovarian epithelial carcinoma 19.4% expressed OPCML mRNA, while 85% of normal ovarian tissue and 76.5% of benign ovarian tumor. The ratio of expression of OPCML mRNA in ovarian epithelial carcinoma was significantly lower than those of normal (chi2 = 30.108, P = 0.0000) and benign tumors (chi2 = 21.162, P = 0.000). No OPCML mRNA expression was found in SKOV-3 and CAOV3, but was found in 3AO. Methylations were detected in 44.4% of cancer cells promoter, while 0% in normal ovarian tissue and benign ovarian tumors. The ratio of methylation of ovarian epithelial carcinoma was significantly higher than those of normal (chi2 = 13.630, P = 0.0000) and benign tumors (chi2 = 11.797, P = 0.000). Methylation was found in SKOV-3 and CAOV3, but not in 3AO. The relationship between gene expression and promoter methylation was correlated (r = 11.589, P = 0.002), especially at Hap I1 site (r = 11.640, P = 0.004). Methylation was also found in SKOV-3 and CAOV3 cell lines, but not in 3AO cell line.</p><p><b>CONCLUSION</b>Deletion of OPCML gene exists in ovarian epithelial carcinoma cell. The gene promoter methylations, especially Hap II motif, may be one of pathways that contribute the inhibition of OPCML expression.</p>

The effects of portal vein microscopic and macroscopic tumor thrombi on post-operation patients with hepatocellular carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the effects of portal vein microscopic and macroscopic tumor thrombi on post-operation patients with hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Three thousand three hundred and forty eight HCC patients were retrospectively reviewed, which were divided into no portal vein tumor thrombi (PVTT), microscopic PVTT and macroscopic PVTT groups according to the pathology, effects of portal vein microscopic and macroscopic tumor thrombi on post-operation patients's survival were studied by univariate analysis and overall survival was evaluated in each group.</p><p><b>RESULTS</b>Hazard ratio (HR) of portal vein microscopic tumor thrombi and macroscopic tumor thrombi was 1.421 and 3.136 respectively; The overall 1-, 3-, 5- and 10-year cumulative survival rate was 85.97%, 62.78%, 49.88% and 35.42% respectively, and mean time for survival was 59.7 months in group without PVTT, while 74.42%, 51.66%, 39.25% and 27.28% respectively and mean time for survival 39.1 months in group with microscopic PVTT, 52.59%, 25.97%, 20.42% and 11.33% respectively and mean time for survival 13.5 months in group with macroscopic PVTT.</p><p><b>CONCLUSIONS</b>PVTT was an important prognostic factor for survival in post-operation patients with HCC while macroscopic PVTT was more danger than microscopic PVTT. The period of microscopic PVTT was the landmark affecting post-operation survival.</p>

Cryohepatectomy for liver cancer: preliminary evaluation of reducing postoperative recurrence / 中华外科杂志

<p><b>OBJECTIVE</b>To determine whether cryohepatectomy is potentially beneficial in reducing the recurrence and prolonging survival for hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>The study included 84 patients who underwent cryohepatectomy, cryosurgery with liquid nitrogen (-196 degrees C) followed by the resection of the frozen tumor by conventional technique, for HCC and were closely follow-up after surgery. Recurrence and survival rates were calculated by the life-table method.</p><p><b>RESULTS</b>The postoperative course of cryohepatectomy in all of the 84 patients was uneventful, there being no operative mortality or severe complications. The 1-, 3-, and 5-year survival rates after cryohepatectomy were 98.7%, 83.9% and 64.0%, respectively. The 1-, 3-, and 5-year recurrence rates after cryohepatectomy were 15.1%, 30.1% and 39.0%, respectively.</p><p><b>CONCLUSIONS</b>Cryohepatectomy for HCC is a safe procedure and may be potentially beneficial in reducing recurrence and prolonging survival. More time is needed to further define whether this procedure will improve long-term survival as compared with conventional resection.</p>

Expressions of VEGF/VEGFRs and activation of STATs in ovarian carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the expressions of VEGF/VEGFRs and activation of STATs in ovarian epithelial carcinoma, and to elucidate direct effect of VEGF on ovarian carcinoma cells.</p><p><b>METHODS</b>Tissue samples from 42 women with primary ovarian epithelial carcinoma (OVCA), 29 with begnin ovarian tumor (OVBT) and 11 with normal ovarian tissue (NOV) were collected. LSAB immunohistochemical staining was used to determine the expression of VEGF, VEGFR1, VEGFR2 and activated STATS (P-STAT1, P-STAT3, P-STAT5, P-STAT6) proteins.</p><p><b>RESULTS</b>(1) Semi-quantitative scoring showed that VEGF expression in OVCA was significantly higher than that in OVBT and NOV (P < 0.01). Expressions of VEGFR1 and VEGFR2 were significantly elevated in OVCA, including tumor cells and stromal vascular endothelial cells (P < 0.01, compared with OVBT and NOV). There was no difference in VEGFRs expressions between OVBT and NOV. (2) In OVCA, tumor cells and endothelial cells expressed P-STAT3 and P-STAT5 at significantly higher levels than those in OVBT and NOV (P = 0.000). The staining of P-STAT1 and P-STAT6 was weak with no significant differences among OVCA, OVBT and NOV. (3) Expressions of VEGFR1 and VEGFR2 in endothelial cells were significantly correlated with P-STAT5 and P-STAT3, respectively (P = 0.006 and 0.001). In cancer cells, VEGF, VEGFR1 and VEGFR2 were all significantly correlated with P-STAT3 and P-STAT5 (P = 0.000), but not with P-STAT1 or P-STAT6.</p><p><b>CONCLUSION</b>VEGF affects ovarian carcinoma cells via VEGFRs, and STATs probably participate in intracellular signaling of VEGF.</p>

Effects of vascular endothelial growth factor on differentiation and function of dendritic cells generated from CD34+ hematopoietic progenitor cells in vitro / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of vascular endothelial growth factor (VEGF) on differentiation and function of dendritic cells derived from CD34+ hematopoietic progenitor cells.</p><p><b>METHODS</b>After isolation from umbilical cord blood with a high-gradient magnetic cell sorting system (MACS), the CD34+ cells were cultured with a cocktail cytokines for differentiating into dendritic cells (DC). The cells were stimulated by VEGF (25 ng/ml) either at the beginning or at day 9 of culture. Kinetics analysis of cell proliferation was performed during the process of cell culture, and the expression of DC differentiation antigens including CD1alpha, CD83, CD80, CD54 and HLA-DR was examined by flow cytometry. DC function was evaluated by the ability to induce proliferation of allogeneic T cells in mixed lymphocyte reaction (MLR) assay, and the production of IL-12 by ELISA.</p><p><b>RESULTS</b>VEGF added at day 1 of culture induced an increase of total cell numbers by (1.51 +/- 0.23)-folds (P = 0.001). VEGF added at the initial but not the late stage of culture could dramatically down-regulate the expression of CD1a [(33.00 +/- 2.12)% vs (81.20 +/- 6.93)%], CD83 [(42.23 +/- 1.15)% vs (87.98 +/- 7.97)%], CD80 (42.93 +/- 1.32)% vs (94.53 +/- 0.87)%], and HLA-DR [(37.93 +/- 5.30)% vs (74.15 +/- 3.74)%], while obviously up-regulate the expression of CD14. Moreover, the inhibitory effect of VEGF on DC function was confirmed by a reduced ability to induce proliferation of allogeneic T cells and production of IL-12 (P < 0.01).</p><p><b>CONCLUSIONS</b>VEGF could induce the expansion of hematopoietic progenitor cells and inhibit at the early stage their differentiation into mature DC.</p>

Role of adiponectin and its receptors in anti-atherosclerotic effects of pioglitazone on ApoE knocked out mice / 北京大学学报(医学版)

Objective:To study the effects of pioglitazone on atherosclerosis on ApoE-/-mice,and to investigate the roles of adiponectin and its receptors.Methods:ApoE-/-mice were fed with high-fat chow for the induction of atherosclerosis and were divided into three subgroups:placebo(n=10),low-dose[10 mg/(kg?d),n=10] pioglitazone therapy,and high-dose[20 mg/(kg?d),n=10] pioglitazone therapy.C57BL/6J wild type mice(n=9) were used as control.Aortic atherosclerosis and intima-media thickness(intima-media thickness,IMT) of abdominal aorta were monitored,and plasma adiponectin was also measured.Expression levels of the adiponectin receptor 1(AdipoR1)and adiponectin receptor 2(AdipoR2) in vessels were analyzed(RT-PCR).Results:(1) Aortic atherosclerotic lesions were observed in ApoE-/-mice but not in wild type mice.Interestingly,these lesions were significantly prevented by high-dose pioglitazone therapy.Compared with wild type mice,ApoE-/-mice had increased IMT of abdominal aorta [(0.290?0.063 vs 0.178?0.012) cm,P

Establishment of methotrexate-resistant human choriocarcinoma cell line and its biological characteristics / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics.</p><p><b>METHODS</b>MTX-resistant cell line (JAR/MTX) was derived from human choriocarcinoma cell line JAR by exposed to intermittently and progressively increasing concentration of MTX. Drug sensitivity was detected by MTT; P-gp GST-Pi and PCNA expressions were detected by immunohistochemistry. Cell apoptosis was detected by flow cytometry (FCM) with PI/Annexin V stain. Growth rates and human chorionic gonadotropin (HCG) production were also measured.</p><p><b>RESULTS</b>JAR/MTX cell line was established with stable MTX-resistance (resistance index to MTX was 7.3) and cross-resistant to TAX and VCR. Growthrate of JAR/MTX was lower than that of parent cell line JAR. Expression level of PCNA in JAR/MTX was lower than that in JAR (3.09+/-0.42 compared with 3.72+/-0.35, P<0.05), while GST-pi expression was higher. No statistical difference of P-gp expression existed between two cell lines. JAR/MTX secreted more HCG than JAR every 10(5) cells secreted (95.7+/-5.4 compared with 41.3+/-2.8)mIU after 48 h(P<0.01). The flow cytometry showed that the spontaneous and MTX induced apoptosis in JAR/MTX was significantly lower than that in JAR P<0.05.</p><p><b>CONCLUSION</b>JAR/MTX cell line presented stable resistant to MTX and cross-resistant to TAX and VCR, which might sever as a model in study of drug resistance in choriocarcinoma.</p>

Activation of signal transducers and activators of transcription induced by vascular endothelial growth factor in CD34+ hematopoietic progenitor cells in vitro / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the activation pattern of signal transducers and activators of transcription (STAT) induced by vascular endothelial growth factor (VEGF) in CD34+ hematopoietic progenitor cells, and gain an insight into the molecular mechanism and signal transduction pathway of VEGF that has an effect on CD34+ hematopoietic progenitor cells.</p><p><b>METHODS</b>After isolated from umbilical cord blood by using a high-gradient magnetically activated cell sorting system (MACS), CD34+ cells were stimulated by VEGF (50 ng/ml) for different time (0, 15, 30, 45, 60, 90 min) to detect the tyrosine phosphorylation and nuclear translocation of STAT-3 and STAT-5 with Western blot and immunocytochemistry methods. The expression of VEGF receptor-2 (VEGFR2) on the membrane of CD34+ progenitor cells was examined by immunocytochemistry. ATWLPPR, an effective peptide screened from phage epitope library by affinity for membrane-expressed VEGFR2 and blocking the binding of VEGF to VEGFR2, was used to determine whether the activation of STAT pathway induced by VEGF was blocked.</p><p><b>RESULTS</b>Tyrosine phosphorylation of STAT-3 and STAT-5 was undetectable in unstimulated CD34+ cells, but was evident at 15 min in response to VEGF stimulation. VEGF resulted in a rapid and transient tyrosine phosphorylation of STAT-3 and STAT-5. The maximal tyrosine phosphorylation was catched at 30 and 45 min, respectively (P = 0.0001), and returned to basal levels at 90 min. Immunocytochemistry confirmed that increased phosphorylated STAT-3 was translocated into the nuclei at 30 min (P = 0.0001), and mainly in cytoplasms again at 90 min after stimulation with VEGF. However, compared with unstimulated CD34+ cells, there was only increased phosphorylation of STAT-5 appeared mainly in cytoplasms, but no significant nuclear translocation was found after stimulation with VEGF (P > 0.05). The presence of VEGFR2 was confirmed using anti-VEGFR2 antibody staining by immunocytochemistry, moreover, the phosphorylation of STAT-3 and STAT-5 failed to be activated by the co-culture with ATWLPPR and VEGF, suggesting that activation of the STAT pathway be specifically mediated by VEGFR2 in CD34+ progenitor cells.</p><p><b>CONCLUSIONS</b>STAT signaling pathway participates in the signal transduction of VEGF via VEGFR2 in CD34+ hemopoietic progenitor cells.</p>

Establishment of two-dimensional electrophoresis of uterine leiomyoma for the proteome analysis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish and optimize the two-dimensional electrophoresis (2-DE) of uterine leiomyoma for the proteome analysis.</p><p><b>METHODS</b>Run immobilized pH gradient (IPG)-isoelectric focusing electrophoresis as the first dimension, then vertical SDS-PAGE electrophoresis as the second dimension. A series of important steps,such as sample solubility, volume of loading, electrophoresis parameters and protocol for staining were optimized.</p><p><b>RESULTS</b>The 2-DE patterns of uterine leiomyoma and myometrium with good quality were obtained.</p><p><b>CONCLUSION</b>With optimal condition the two-dimensional electrophoresis of uterine leiomyoma can be obtained.</p>

Effect of postoperative transcatheter arterial chemoembolization on hepatocellular carcinoma patients with residual tumor / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the effect of postoperative adjuvant transcatheter arterial chemoembolization (TACE) on hepatocellular carcinoma (HCC) patients with residual tumor.</p><p><b>METHODS</b>The patients were classified into intervention group (with adjuvant TACE) and control group (without adjuvant TACE) who were further stratified to those with high risk (patients with single tumor > 5 cm in diameter, or with multiple tumors, invasion to blood vessels), and low risk factors. Univariate analysis and Cox model were used to analyse prognostic factors.</p><p><b>RESULTS</b>In low risk patients with residual tumor, the 1-, 2-, 3-, 4-year survival rate was 97.2%, 78.0%, 66.5% and 66.5% in the intervention group, and 91.2%, 81.4%, 70.3% and 54.4% in the control group, respectively. There was no statistical difference between the two groups in survival (log-rank P = 0.7667). Comparing with the control group, the 1-, 2-, 3-, 4-year survival rate was 89.5%, 73.4%, 59.2% and 53.8% in the intervention group, and 70.5%, 61.9%, 46.8% and 46.8% in the control group, respectively. Postoperative adjuvant TACE significantly prolonged the survival in high risk patients with residual tumor (P = 0.0029). Cox model revealed that the benefit of adjuvant TACE was significantly increased by the high risk factors in HCC patients with residual tumor.</p><p><b>CONCLUSION</b>The beneficial effect of postoperative TACE was only observed in high risk patients with residual tumor but not in the low risk patients with residual tumor.</p>

Three-grade criteria of curative resection for primary liver cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To clarify three-grade criteria of curative resection for primary liver cancer (PLC) and evaluate their clinical significance.</p><p><b>METHODS</b>Criteria of curative resection of PLC were summed up to three grades. Grade I: complete removal of all gross tumors with no residual tumor at the excision margin. Grade II: on the basis of Grade I, there was no extrahepatic metastasis, no hilar lymph node metastasis, no tumor thrombus in the main trunks and their primary tributaries of the portal vein, common hepatic duct, hepatic vein and vena cava inferior, and the tumor was not more than two in number. Grade III: in addition to the above criteria, AFP dropped to normal level (in patients with elevated AFP before surgery) within 2 months after operation, and no residual tumor upon diagnostic imaging. A total of 354 cases with PLC who had their liver resected was reviewed. Patients in each grade were divided into two portions depending on whether the treatment was curative or palliative.</p><p><b>RESULTS</b>The survival of patients receiving curative treatment was better than those receiving palliative treatment (P < 0.01). This was true for patients whose treatment belonged to anyone of the three-grade criteria. The survival was improved along with the promotion of curative criteria used. The 5-year survival rate of Grade I, II and III patients undergone curative resection was 43.2%, 51.2% and 64.4%, respectively (P < 0.01).</p><p><b>CONCLUSION</b>1. The three-grade criteria may be used for judging the radicality of tumor resection for PLC. 2. The more stringent the criteria used, the better the survival would be. 3. Adopting high-grade criteria to select cases, to guide operation and postoperative follow-up would improve the results of liver resection for PLC.</p>

Single methotrexate chemotherapy for low-risk gestational trophoblastic tumor / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the efficacy and toxicity of methotrexate (MTX) give intravenously in the primary treatment of gestational trophoblastic tumor (GTT).</p><p><b>METHODS</b>A total of 37 patients with low-risk GTT was primarily treated by single MTX in Women's Hospital, School of Medicine, Zhejiang University. Data on the patients' age, clinical stage, WHO classification criteria, antecedent pregnancy, presenting level of human chorionic gonadotropin, courses of chemotherapy required to achieve complete remission, and toxicity related to chemotherapy treatments were collected.</p><p><b>RESULTS</b>Thirty-seven patients with low-risk GTT totally received 137 cycles of MTX between Oct. 1999 and Sep. 2002, 34 patients (91.9%) achieved complete remission. Twenty-nine patients received multiple courses of MTX, complete remission was induced in 26 patients (89.7%). The complete response rates of I stage and III stage were 100.0% and 70.0% (P = 0.03) respectively in patients who were received multiple courses of MTX. However, eight patients received single course of chemotherapy, 7 patients achieved complete remission, and 1 achieved complete remission after another additional course of MTX was conducted. Grade III side effects (WHO criteria) only appeared in 7 courses (5.1%) during MTX treatment. Follow-up data showed that only one patient with single course of chemotherapy relapsed after 6 months.</p><p><b>CONCLUSION</b>Single MTX chemotherapy may be effective and well tolerated for low-risk GTT.</p>

Mismatch repair gene promoter methylation and expression in hydatidiform moles and the malignant transformation / 中国医学科学院学报

<p><b>OBJECTIVE</b>In this study, we assayed promoter hypermethylation and protein expression of the mismatch repair gene (MMR) hMLH1 and hMSH2 in gestational trophoblastic diseases to understand the significance of MMR promoter methylation and expression in the pathogenesis and malignant transformation of hydatidiform mole.</p><p><b>METHODS</b>DNA was extracted from chorion of early pregnancies, partial hydatidiform moles, complete hydatidiform moles, and invasive moles were over digested by methylation sensitive endonuclease Hpa II. Then the promoters were amplificated by polymerase chain reaction. The protein was detected by immunohistochemistry.</p><p><b>RESULTS</b>In the normal placenta, neither hMLH1 nor hMSH2 promoter methylation was detected. Expression of hMLH1 and hMSH2 in cytotrophoblasts was strongly positive, and that was negative or weakly positive in syncytiotrophobasts. In all normal chorion, expression of hMLH1 and hMSH2 in cytotrophoblasts was strongly positive. In partial hydatidiform mole and complete hydatidiform mole, the methylation of hMLH1 and hMSH2 promoters was significantly higher than that of early placenta (P < 0.05), and the protein expression in cytotrophoblasts was significantly lower (P < 0.05). In the invasive mole, hMLH1 and hMSH2 promoter methylation were not significantly different as compared with the partial hydatidiform mole and complete hydatidiform mole (P > 0.05). Expression of hMLH1 in the invasive mole (54.5%, 6/11) was not significantly different as compared with the partial hydatidiform mole and complete hydatidiform mole (P > 0.05). But expression of hMSH2 in the invasive mole (36.4%, 4/11) was weaker than that in complete hydatidiform mole (P = 0.044). Promoter methylation and less expression of hMSH2 had correlations in complete hydatidiform mole or invasive mole.</p><p><b>CONCLUSIONS</b>Strong expressions of hMLH1 and hMSH2 in the cytotrophoblasts of normal placenta may keep the genome stability. Promoter methylation and down-regulation of hMLH1 and hMSH2 are probably involved in the pathogenesis of hydatidiform mole.</p>

Hypermethylation of hMLH1 and microsatellite instability in ovarian mucinous tumors / 中国医学科学院学报

<p><b>OBJECTIVES</b>To investigate the role of hMLH1 promoter hypermethylation and microsatellite instability (MSI) in the development of ovarian mucinous tumors.</p><p><b>METHODS</b>One hundred and seven of paraffin-embedded specimens of ovarian mucinous tumors (malignant 49, borderline 35, and benign 23) were collected from Women's Hospital, School of Medicine, Zhejiang University from 1995 to 2001. The assessment of MSI was based on the use of a panel of six microsatellite markers (BAT-25, BAT-26, BAT-40, D5S346, D17S250, and D2S123) by polymerase chain reaction (PCR). Hypermethylation of hMLH1 promoter region was detected using restriction cut analysis.</p><p><b>RESULTS</b>4.3% (1/23), 14.3% (5/35), and 36.7% (18/49) of benign tumors, borderline tumors, and malignant tumors respectively displayed hypermethylation of the hMLH1 promoter. The hMLH1 promoter hypermethylation rate of malignant group was significantly higher than that of borderline and benign group (P = 0.023, 0.004), but no significant difference between the borderline group and the benign group (P = 0.438); 4.3% (1/23), 8.6% (3/35), and 16.3% (8.49) of benign tumors, borderline tumors, and malignant tumors showed MSI positive phenotype. But there were no significant differences each other in the MSI positive phenotype rate; 75% (9/12) MSI positive phenotype ovarian mucinous tumors were hypermethylated at hMLH1 promoter, while the MSI-phenotype tumors were unmethylated in 84.2% (80.95) of cases. There was significant correlation between MSI positive phenotype and hMLH1 promoter hypermethylation (P = 0.000).</p><p><b>CONCLUSIONS</b>In ovarian mucinous tumors, malignant, borderline, and benign tumors exist hMLH1 promoter hypermethylation. Hypermethylation of hMLH1 promoter results MSI in ovarian mucinous tumors. Methylation of hMLH1 promoter and MSI may be involved in the carcinogenesis of ovarian mucinous cancer.</p>

Biologic characteristics of intraperitoneal transplantation model of human ovarian carcinoma in severe combined immunodeficiency mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To develop a human ovarian carcinoma SKOV3 model in severe combined immunodeficiency (SCID) mouse and to study its biologic characteristics.</p><p><b>METHODS</b>Human ovarian carcinoma SKOV3 cells were injected intraperitoneally into female SCID mouse to establish a transplantation model of human ovarian carcinoma. The biological characteristics, metastasis and morphology of transplanted tumors were studied.</p><p><b>RESULT</b>All tumors grew progressively with no sign of regression. The tumor cells spread around the peritoneal cavity and mainly on the diaphragm, mesentery, peritoneum and around the liver, which was confirmed by histopathology. The morphology, growth pattern and CA125 secretion of primary culture of transplanted cells remained as same as those of ovarian carcinoma cell line SKOV3.</p><p><b>CONCLUSION</b>An intraperitoneal transplantation model of human ovarian carcinoma SKOV3 in SCID mice has been developed successfully, which can simulate the biological behavior of peritoneal metastasis of human ovarian carcinoma.</p>