Circulating MicroRNAs in gynecological malignancies: from detection to prediction.

MicroRNAs (miRNAs) have been demonstrated to play critical roles in many physiological and pathophysiological processes. The presence of altered miRNA profiles in human body fluids has been reported for a number of diseases including gynecological malignancies. In this review, we summarized the current progresses of circulating miRNAs associated with malignancies in gynecology, with an emphasizing on the circulating miRNAs as diagnostic and prognostic biomarkers in ovarian cancer, endometrial carcinoma and cervical cancer.

Differentially expressed microRNAs in the serum of cervical squamous cell carcinoma patients before and after surgery.

BACKGROUND: The purpose of this study was to detect the serum microRNAs (miRNAs) that are differentially expressed in cervical squamous cell carcinoma (SCC) patients and negative controls, with a focus on the miRNA profiles of the patients before and after surgery. The aim of the study is to evaluate the potential of these miRNAs as novel markers for the post-therapeutic monitoring of cervical SCC patients. RESULTS: A total of 765 serum miRNAs from 10 cervical SCC patients before surgery, 10 cervical SCC patients after surgery, and 10 negative controls were profiled using a TaqMan MicroRNA Array. A set of selected differentially expressed miRNAs were further analyzed in the patients at different perioperative periods, including preoperative, 1 week postoperative, and one month postoperative. The results showed that several serum miRNAs were differentially expressed in the cervical SCC patients compared with the negative controls, including miR-646, miR-141* and miR-542-3p. More importantly, we found that levels of specific serum miRNAs were deregulated in the pre- and postoperative stages, and these miRNAs could be useful for post-therapeutic monitoring of disease progression. Finally, we depicted a regulatory network of differentially expressed serum miRNAs, and many possible target genes were predicted in the estrogen-mediated signal pathways, supporting the hypothesis that cervical SCC is a hormone-associated gynecological disease. CONCLUSIONS: Our study demonstrated that the circulating miRNAs miR-646, miR-141* and miR-542-3p could potentially serve as non-invasive biomarkers for cervical SCC. The levels of these specific miRNAs might be useful for the post-therapeutic monitoring of disease progression. This is the first report showing that circulating miRNAs could serve as biomarkers for the therapeutic intervention of cervical SCC.

Circulating microRNAs identified in a genome-wide serum microRNA expression analysis as noninvasive biomarkers for endometriosis.

CONTEXT: There is currently no reliable noninvasive biomarker for the clinical diagnosis of endometriosis. Previous analyses have reported that circulating microRNAs (miRNAs) can serve as biomarkers for a number of diseases. OBJECTIVE: The study aims to detect the serum miRNAs that are differentially expressed between endometriosis patients and negative controls to evaluate the potential of these miRNAs as diagnostic markers for endometriosis. DESIGN: A total of 765 serum miRNAs were profiled using a TaqMan microRNA array in a pool of 10 endometriosis patients and a pool of 10 negative controls, and a set of selected miRNAs were further analyzed in a validation cohort consisting of sera from 60 patients and 25 controls including 10 samples used in array profiling. RESULTS: The relative expression levels of miR-199a and miR-122 were found to be up-regulated in endometriosis patient samples compared with control samples, whereas miR-145*, miR-141*, miR-542-3p, and miR-9* were down-regulated in endometriosis patients. Importantly, the relative expression of miR-199a (P < 0.05) and miR-122 can be used to discriminate between severe and mild endometriosis. We also found that miR-199a is well correlated with pelvic adhesion and lesion distribution (P < 0.05) and associated with hormone-mediated signaling pathways. Furthermore, we investigated the diagnostic value of these molecules and confirmed the optimal combination of miR-199a, miR-122, miR-145*, and miR-542-3p with area under the curve of 0.994 (95% confidence interval = 0.984-1.000, P < 0.001) and a cutoff point (0.4950) of 93.22% sensitivity and 96.00% specificity. CONCLUSIONS: Our study demonstrated that the circulating miRNAs miR-199a, miR-122, miR-145*, and miR-542-3p could potentially serve as noninvasive biomarkers for endometriosis. miR-199a may also play an important role in the progression of the disease. This is the first report that circulating miRNAs serve as biomarkers of endometriosis.

Expression of Bmi-1, P16, and CD44v6 in Uterine Cervical Carcinoma and Its Clinical Significance.

OBJECTIVE: Bmi-1, a putative proto-oncogene, is a core member of the polycomb gene family, which is expressed in many human tumors. The p16 protein negatively regulated cell proliferation, whereas CD44v6 is associated with proliferation as an important protein. Additionally, CD44v6 is an important nuclear antigen closely correlated to tumor metastasis. The present study aims to investigate the expression and significance of Bmi-1, p16, and CD44v6 in uterine cervical carcinoma (UCC). METHODS: A total of 62 UCC, 30 cervical neoplasic, and 20 normal cervical mucosal tissues were used in the current study. The expression of Bmi-1, p16, and CD44v6 in these tissues was determined using immunohistochemical assay. The relationships among the expression of these indices, the clinicopathologic features of UCC, and the survival rate of UCC patients were also discussed. The correlation between Bmi-1 protein expression and p16 or CD44v6 protein in UCC was analyzed. RESULTS: The expression of Bmi-1, p16, and CD44v6 was significantly high in cervical carcinoma compared with that in the cervical neoplasia and normal colorectal mucosa (P<0.05). The over-expression of Bmi-1 protein in UCC was apparently related to the distant metastasis (P<0.01) and the tumor, nodes and metastasis-classification, i.e. the TNM staging, World Health Organization (P<0.05). Nevertheless, the positive expression of p16 protein in UCC was not significantly associated with the clinicopathologic features (P>0.05). The Kaplan-Meier survival analysis showed that the over-expression of Bmi-1 significantly decreased the survival rate of UCC patients (P<0.05). A strong correlation indicated that there was statistical significance between the expression of Bmi-1 and CD44V6 proteins in UCC (r=0.419, P=0.001). CONCLUSIONS: The over-expression of Bmi-1 and CD44v6 protein closely correlate to the tumorigenesis, metastasis, and prognosis of UCC. Bmi-1 and CD44v6 may be used to predict the prognosis of cervical carcinoma. Bmi-1 may indirectly regulate the expression of CD44v6 in UCC patients. The positive expression of p16 protein is possibly associated with the tumorigenesis, but not with the metastasis or prognosis of UCC.

Clinical Significance of CENP-H Expression in Uterine Cervical Cancer.

OBJECTIVE: This work aims to investigate the expression pattern and clinicopathologic significance of centromere protein H (CENP-H) in uterine cervical cancer (UCC). METHODS: The level of CENP-H expression in the paraffin sections of 62 UCC cases was determined by the SP immunohistochemical method, with complete clinicopathologic data in all cases. Statistical analysis was conducted to evaluate the prognostic and diagnostic significance of CENP-H using SPSS13.0 software package. RESULTS: Immunohistochemical assay showed strong CENP-H expression in 61.29% (38/62) of the paraffin-embedded cervical cancer tissues. Statistical analysis revealed a strong correlation between the CENP-H expression and the clinical classification (P=0.038) of the cervical carcinoma. The expression increased with rise of the stages. The analysis of Cox proportional hazards regression model suggested that CENP-H expression (P=0.002) and tumor stage (P=0.001) were independent prognostic markers for the survival of UCC patients. The survival analysis showed that the survival rate was significantly lower in patients with high expression of CENP-H than in those with low expression of CENP-H (P=0.001). CONCLUSIONS: CENP-H is likely to be a valuable marker for carcinogenesis and progression of UCC. It might be used as the important diagnostic and prognostic marker for cervical carcinoma patients, especially for those at early stage.

MRI appearance of dysembryoplastic neuroepithelial tumor / 中华神经医学杂志

Objective To study the MRI appearance and clinical features of dysembryoplastic neuroepithelial tumor (DNT) to improve accurate diagnosis of DNT. Methods The clinical data and MRI appearance of 12 patients with DNT confirmed by surgery and pathology were analyzed retrospectively. Routine MRI was performed in all of the 12 patients, and also dynamic contrast-enhanced MR imaging in 10 of them. Results Eight lesions resided in the temporal lobes, 2in the frontal lobe, and the other 2 in the parietal lobe. All of the 12 DNTs located in supratentorial hemisphere cortex, and 6 of them encroached the adjacent white matter. The lesions appeared as hypointense or iso-hypointense signal on T1WI of MRI, and hyperintense on T2WI and FLAIR of MRI.Cystic structure was shown in all the lesions or as its main part, and mesh-separated structure was also noted in some of the tumors. Five lesions appeared as a triangle in shape, 5 in gyrus-like shape and the remaining 2 in round shape. All the tumors had no significant mass effect with 1 having mild edema around the tumor. Enhanced MR imaging showed only 1 lesion having slight and heterogeneous enhancement. Conclusion DNT is accorded with general tumor's biological characteristics. MRI has great value on DNT's diagnosis. If a cystic lesion locates in the hemisphere cortex with a triangle in shape or gyrus-like shape, having no significant mass effect and peripheral edema, it has a great possibility of being DNT.

MRI diagnosis of central neurocytoma / 中华神经医学杂志

Objective To evaluate the MRI features of central neurocytoma (CNC). Methods Thirteen patients with CNC (7 males and 6 females, aged 18-28), admitted to our hospital fiom January 2007 to January 2010, were chosen; preoperative plain and contrast enhanced MRI scans were performed on these 13 patients and 1 also received CT scan. The clinical and imaging data and pathological features of these patients were retrospectively analyzed. Results The largest tumor enjoyed its maximum diameter of 3.2-8.5 cm; the CNC in 12 patients located at the lateral ventricle and that in 1 patient at the parenchyma of the left frontal and temporal lobes. Lobulated solid tumors were noted in these lesions,showing T1 isointense and T2 hyperintense in MRI; some multiple, scattered cystic lesions showed stronger T1 and T2 signal than the former ones; vascular "flow void" signal was shown in 6 patients in T2WI, and slightly weak T1 signal was shown in 8. All the tumors demonstrated inhomogeneous enhancement and the tumor vessels in 6 patients were noted under contrast MRI. Focal calcification was found. Conclusion CNC in the lateral ventricle enjoys typical location; considering the age of patients, non-contrast and contrast MRI findings, correct diagnosis of CNC can be made; however, CNC outside the lateral ventricle is hard to diagnose.

Cumulus cells and their extracellular matrix affect the quality of the spermatozoa penetrating the cumulus mass.

OBJECTIVE: To investigate the role of the cumulus cells and the cumulus matrix in affecting the penetrability, morphology, acrosome reaction, and motility of human spermatozoa penetrating the cumulus oophorus. DESIGN: Controlled experimental laboratory study. SETTING: University gynecology unit. PATIENT(S): Women undergoing assisted reproduction treatment and men visiting the subfertility clinics. INTERVENTION(S): Human spermatozoa were allowed to penetrate through the cumulus oophorus and cell-depleted cumulus matrix in a capillary, and were treated with cumulus cell extract or hyaluronic acid. MAIN OUTCOME MEASURE(S): The morphology, acrosomal status, and motility of human spermatozoa were determined. RESULT(S): Fewer spermatozoa could penetrate the fresh cell-depleted matrix compared with intact cumulus oophorus. Spermatozoa that penetrated through the cumulus oophorus had higher percentages of normal morphology and acrosome reaction and had specific motility pattern. These effects were lost or reduced in the cell-depleted matrix that had been stored overnight. Hyaluronic acid, a main component of the cumulus matrix at concentration found in the cumulus oophorus, modulated sperm motility but did not affect spontaneous acrosome reaction. Cumulus cell extract did not affect sperm motility, but induced acrosome reaction. CONCLUSION(S): Both the cumulus matrix and the cumulus cells contribute to the effect of cumulus oophorus on spermatozoa penetrating through it.

Application of virtual imaging technique in diagnosis of intracranial aneurysms / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the value of virtual imaging technique in diagnosis of intracranial aneurysms.</p><p><b>METHODS</b>Fifty-four cases of 54 intracranial aneurysm diagnosed by three-dimensional CT angiography (3D-CTA) examinations were enrolled in this study. Three-dimensional virtual images of the skull and cerebral vessels were acquired by three-dimensional reconstruction of the original CT images using the surgical planning system, and the location, size and shape of the aneurysms and their anatomical relationship with the adjacent tissues were observed and measured from several angles. All the patients underwent surgical planning and simulated surgical operations using the virtual surgical instruments available in the system.</p><p><b>RESULTS</b>All the 54 cases had successful three-dimensional virtual image reconstruction and the surgical planning operations. The virtual imaging system generated clear and vivid three-dimensional virtual images which clearly visualized the location and size of the aneurysms and their precise anatomical relations to the parent arteries and skull. This virtual reality imaging system also allowed simulation of simple surgical procedures.</p><p><b>CONCLUSION</b>The surgical planning system based on the virtual reality imaging can serve as a useful means to assist the diagnosis and provide precise imaging details of intracranial aneurysms.</p>

Combined application of virtual imaging techniques and three-dimensional computed tomographic angiography in diagnosing intracranial aneurysms / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The diagnostic value of virtual imaging combined with three-dimensional computed tomographic angiography (3D-CTA) for intracranial aneurysms has not been fully elucidated yet. This study aimed to evaluate the value of combined application of virtual imaging techniques and 3D-CTA in diagnosing patients with aneurismal subarachnoid hemorrhage (SAH) at the acute stage.</p><p><b>METHODS</b>Eighty patients with non-traumatic SAH received 3D-CTA examinations. The raw CT data of these patients were reconstructed and transferred into the 3D mode through the surgical plan system based on virtual reality (VR) image, and the 3D virtual images of skulls and brain blood vessels were acquired. The location, size and shape of aneurysms and their anatomic relationship with adjacent tissues were measured from many points of view.</p><p><b>RESULTS</b>Seventy-three aneurysms were detected in 68 of the 80 patients, but 2 aneurysms were detected in 2 of the 5 patients who had been found free of aneurysms previously and had received 3D-CTA examinations for a second time one month later. The 3D virtual images produced by the virtual imaging system were clear and vivid, and they could reveal the location and size of the aneurysm and its relations to the parent artery and skull directly.</p><p><b>CONCLUSIONS</b>The imaging of 3D-CTA is convenient, reliable and fast in diagnosing intracranial aneurysms and can be regarded as the first choice for the diagnosis and treatment of ruptured intracranial aneurysms. Combined with the surgical plan system based on the VR image, 3D-CTA may obtain more imaging information about aneurysms.</p>

Drug sensitivity test and drug-resistance gene assessment for chemotherapy planning for tumors derived from high-passage mouse gastric cancer cells: therapeutic effect observation in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of chemotherapy with drugs selected according to drug sensitivity test and drug- resistance gene assessment in mice bearing tumors derived from high-passage gastric cancer cells.</p><p><b>METHODS</b>Chemosensitivity of the mouse gastric cancer cells to 6 chemotherapeutic drugs was investigated using MTT assay before the 6 drugs were applied in mice with implanted tumors. The implanted tumor volume and tumor inhibition rate were observed, and the expression of the multidrug resistance-1 (MDR1) gene was detected.</p><p><b>RESULTS</b>5-FU, the most sensitive among the 6 drugs, resulted in significantly higher tumor inhibition rates in comparison with the control group, but did not significant affect MDR1 expression. Adriamycin (ADM) had the lowest sensitivity with low tumor inhibition rates and no significant effect on MDR1 expression. Cisplatin (DDP) was identified as the most sensitive drug for mouse gastric cancer with previous 5-FU exposure, and Mitomycin (MMC) was the most insensitive drug for mouse gastric cancer with previous ADM treatment. In mice bearing tumors of the second-passage cells, DDP showed high tumor inhibition rate but scarcely affected MDR1 expression; MMC resulted in low tumor inhibition rate and high MDR1 expression. Application of 5-FU and ADM in these tumor-bearing mice of the second-passage cancer cells resulted in rather low tumor inhibition rates without significant alterations in MDR1 expression as compared with the control group.</p><p><b>CONCLUSION</b>Appropriate individualized chemotherapy should be implemented in full consideration of the results of drug sensitivity test and drug-resistance gene assessment.</p>

Cumulus cells reduce the spermatozoa-zona binding inhibitory activity of human follicular fluid.

OBJECTIVE: To investigate the effects of human follicular fluid cultured with cumulus cells to inhibit the binding of spermatozoa to the zona pellucida of oocytes. DESIGN: Controlled experimental laboratory study. SETTING: University gynecology unit. PATIENT(S): Women undergoing assisted reproduction program and men visiting the subfertility clinics. INTERVENTION(S): Culture medium and human follicular fluid were used to culture cumulus cells in vitro for specified time periods. MAIN OUTCOME MEASURE(S): Zona binding capacity and motility of spermatozoa after incubation with cumulus cells treated culture medium or human follicular fluid. RESULT(S): Compared with the control medium, spent culture media after culturing cumulus cells for 3, 5, and 7 hours did not affect the motility and zona binding capacity of the treated spermatozoa. Significantly more spermatozoa treated with human follicular fluid that had been preincubated with cumulus cells for 5 and 7 hours bound onto hemizona in hemizona binding assay when compared with those preincubated in human follicular fluid without cumulus treatment. The hemizona index increased with the increase in the duration of cumulus cell treatment. Human follicular fluid with or without cumulus cells maintained sperm motility to similar extent for 3 hours. CONCLUSION(S): Cumulus cells reduced the inhibitory effect of human follicular fluid on spermatozoa-zona binding in vitro in a time-dependent manner.