Histological evaluation of cervical carcinomas in FIGO stage Ib2/IIa after neoadjuvant chemotherapy / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the histological changes of cervical cancer after neoadjuvant chemotherapy (NACT) and to establish histological criteria for interpretation of chemotherapeutical effects.</p><p><b>METHODS</b>Fifty-six patients with FIGO stage Ib2-IIa cervical cancers treated by NACT and subsequent radical surgery were retrospectively analyzed, in which the pre- and post-chemotherapeutic histopathological changes were assessed.</p><p><b>RESULTS</b>The post-chemotherapeutic histopathological changes of 56 cases included grade 3 effects in 11 cases (19.6%), grade 2 in 24 cases (42.9%), grade 1 in 13 cases (23.2%) and no response in only 8 cases (14.3%). The histologic response rate was 62.5% (35/56) and the overall clinical response rate was 67.9% (38/56). The overall coincidence by both criteria was 78.6% (44/56). Four cases (7.1%, 4/56) had only histological response and 8 cases (14.3%, 8/56) had response by imaging. In comparison with the pre-chemotherapy specimens, the chemotherapy-associated histological changes included shrinkage and scattering of tumor nests,decrease of tumor cellularity,tumor cell degeneration and necrosis.</p><p><b>CONCLUSIONS</b>The histological changes in locally advanced cervical cancers induced by NACT are significant, which may challenge the diagnosis in the final specimens. There are some discreqancies between the histological criteria and imaging/gynecological ones for the therapeutic evaluation of cervical cancers,and it is thus recommended to use the pathological criteria for clinic practice.</p>

Value of P16 expression in the triage of liquid-based cervical cytology with atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesions / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The management of atypical squamous cells of undetermined significance/low-grade squamous intraepithelial lesions (ASCUS/LSIL) is still controversial and it is advisable to make a triage for these two cytological abnormalities. P16(INK4) (P16) has been shown to be a potential biomarker for predicting high-grade cervical intraepithelial neoplasia (CIN) and cervical cancer. The aim of the study was to determine the value of P16 expression by immunostaining method compared with high-risk human papillomavirus (HR-HPV) DNA test in the triage of ASCUS/LSIL women.</p><p><b>METHODS</b>Totally 86 eligible residual liquid-based cytological specimens with ASCUS and 45 with LSIL were obtained. All specimens were submitted to HR-HPV DNA test (HC2) and P16 immunocytochemical staining simultaneously. And all women underwent colposcopy and biopsy after cytology.</p><p><b>RESULTS</b>The positive rate of P16 staining was 32.6% in ASCUS and 42.2% in LSIL, which was significantly lower than that of HR-HPV test in both ASCUS (P < 0.05) and LSIL (P < 0.05). Moreover, the positive rate of P16 staining was 12.7% in normal histology, 61.5% in CIN 1, 87.0% in CIN 2-3, and 100.0% in cancer, in which P16 positive rate was significantly lower than HR-HPV positive rate in normal group. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of P16 staining for predicting CIN 2 or more were 87.5%, 68.6%, 38.9%, 96.0%, and 72.1%, respectively in the ASCUS; while 90.0%, 71.4%, 47.4%, 96.2% and 54.7%, respectively in the LSIL, in which the specificity and accuracy of P16 staining were significantly higher than those of HR-HPV test in both ASCUS and LSIL (P < 0.05).</p><p><b>CONCLUSION</b>P16 immunostaining had significantly higher specificity and accuracy than HR-HPV DNA test for predicting for high-grade CIN and cervical cancer in ASCUS and LSIL and can be used for the triage of women with ASCUS/LSIL cytological abnormality.</p>

Anti-fertility effect of Tongbi composition and its reversibility in male rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the anti-fertility effect of maximum-dose Tongbi Composition and its reversibility in male rats.</p><p><b>METHODS</b>Thirty-six male SD rats were equally randomized into a control group and a medication group, the former given normal saline at 10 ml/(kg x d), while the latter treated with Tongbi Composition at 10 g/(kg x d), both for 60 days. Half the rats of each group were sacrificed randomly at the cessation of treatment, and the rest killed at 72 days after it. The relative testis weight, testis volume, sperm concentration and sperm motility were measured, and the pathological changes in the testicular tissue observed under the optical microscope.</p><p><b>RESULTS</b>After 60 days of treatment, no statistically significant differences were found between the two groups in the relative testis weight, testis volume and sperm concentration (P > 0.05) , and the sperm motility of the medication group dropped to zero, but it was restored to normal at 72 days after drug withdrawal. Almost no lesions were observed in the testis tissue of the medication group.</p><p><b>CONCLUSION</b>The short-term use of Tongbi Composition at the maximum clinical dose has an obvious anti-fertility effect, but it is reversible.</p>

Accuracy of tumor grade by preoperative curettage and associated clinicopathologic factors in clinical stage I endometriod adenocarcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Preoperative tumor grading becomes one of the most important predictors for lymphadenectomy at primary surgery for clinical stage I endometriod adenocarcinoma. However, there is an inconsistency of tumor grade between preoperative curettage and final hysterectomy specimens, and its associated factors are poorly understood. This study aimed to evaluate the accuracy of tumor grade by preoperative curettage so as to achieve a better stratified management for clinical stage I endometriod adenocarcinoma.</p><p><b>METHODS</b>Clinical data of totally 687 patients with clinical stage I endometriod adenocarcinoma who underwent preoperative curettage and primary surgery were retrospectively collected. Compared with final hysterectomy specimens, the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of tumor grade by preoperative curettage were calculated and their associations with clinicopathologic parameters, including age, status of menopause, position of uterus, location and size of lesion, histological grade, depth of myometrial invasion, cervical invasion, extrauterine spread, peritoneal cytology, metastasis to retroperitoneal lymph node, serum CA125 level, and hormone receptor status, were analyzed.</p><p><b>RESULTS</b>In final hysterectomy specimens, 139 of 259 grade 1 patients by curettage were upgraded to grade 1 or 2; 31 of 296 grade 2 were upgraded to grade 3, with a significantly discrepant rate of 40.9% (281/687) and an upgraded rate of 24.7% (170/687). The specificity and negative predictive value for grade 3 were 90.7% and 89.9%, while the sensitivity and positive predictive value for grade 1 were 67.1% and 40.9%, respectively.</p><p><b>CONCLUSIONS</b>Preoperative tumor grade by curettage does not accurately predict final histological results, especially in those classified as grade 1. Complete surgical staging seems to be necessary for clinical stage I endometriod adenocarcinoma.</p>

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>

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>

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>

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>

Molecular cloning of human IL-7cDNA and construction of eukaryotic vector expressing hIL-7 / 浙江大学学报·医学版

OBJECTIVE: To construct a vector expressing eukaryotic human interluken-7(hIL-7). METHODS: hIL-7 DNA was identified and cloned (cDNA) from human spleen tissue using reverse transcription polymerase chain reaction (RT-PCR). We incorporated the cDNA into the pMD18-T plasmid. The pMD18-T plasmid was then inserted into a dual expression vector (prokaryotic and eukaryotic) pBK-CMV and called pBK-CMV-hIL-7. We used pBK-CMV-hIL-7 vector to infect E.coli DH5alpha. The expression of the recombinant hIL-7 protein (rhIL-7) by E.coli DH5alpha was analyzed using SDS-PAGE and western blot testing. RESULTS: The genetically engineered E.coli DH5alpha did express rhIL-7 confirmed by western blot. CONCLUSION: The successful construction of genetically engineered eukaryotic gene for hIL-7 was done, This will enable further research into therapeutic uses for hIL-7.

Expression of mRNA isoforms of vascular endothelial growth factor in ovarian carcinoma / 浙江大学学报·医学版

OBJECTIVE: To study the expression of vascular endothelial growth factor (VEGF) mRNA isoforms in ovarian carcinoma and to explore their role in tumorigenesis and development of ovarian carcinoma. METHODS: The types and levels of VEGF mRNA isoforms of surgical samples from 30 patients with ovarian carcinoma were determined by relatively quantative RT-PCR, nest PCR and sequence analysis. RESULTS: VEGF(121), VEGF(145), VEGF(165) and VEGF(189)mRNA were detected in normal ovaries and ovarian carcinoma tissues. The expression level of VEGF(121) was significantly higher than that of VEGF(145), VEGF(165) and VEGF(189) (P<0.001, respectively). The expression of all 4 isoforms in carcinoma tissues was increased significantly compared with that in normal ovaries (P<0.05). CONCLUSION: Overexpression of VEGF(121), VEGF(145), VEGF (165) and VEGF(189) mRNA, especially VEGF(121), was found in varian carcinoma tissues. This findings suggest that all 4 VEGF isoforms may be involved in the tumorigenesis and development of ovarian carcinoma and VEGF(121) may play a key role.