Generation of fully pluripotent female murine-induced pluripotent stem cells.

The high quality of induced pluripotent stem cells (iPSCs) has been determined to be high-grade chimeras that are competent for germline transmission, and viable mice can be generated through tetraploid complementation. Most of the high-quality iPSCs described to date have been male. Female iPSCs, especially fully pluripotent female iPSCs, are also essential for clinical applications and scientific research. Here, we show, for the first time, that a gender-mixed induction strategy could lead to a skewed sex ratio of iPSCs. After reprogramming, 50%, 70%, and 90% female initiating mouse embryonic fibroblasts at different male ratios resulted in 14.1 ± 6.8% (P < 0.05), 31.8 ± 5.4% (P < 0.05), and 80.1 ± 2.8% (P < 0.05) female iPSCs, respectively. Furthermore, these female iPSCs had pluripotent properties typical of embryonic stem cells. Importantly, these fully pluripotent female iPSCs could generate viable mice by tetraploid complementation. These findings indicate that high-quality female iPSCs could be derived effectively, and suggest that clinical application of female iPSCs is feasible.

Feasibility and Diagnostic Accuracy for Assessment of Coronary Artery Stenosis of Prospectively Electrocardiogram-gated High-pitch Spiral Acquisition Mode Dual-source CT Coronary Angiography in Patients with Relatively Higher Heart Rates: in Comparison with Catheter Coronary Angiography.

Objective To prospectively investigate the diagnostic accuracy for coronary artery stenosis of prospectively electrocardiogram-triggered spiral acquisition mode (high pitch mode) dual-source computed tomography coronary angiography (CTCA) in patients with relatively higher heart rates (HR) compared with catheter coronary angiography (CCA). Methods Forty-seven consecutive patients with relatively higher HR (>65 and <100 bpm) (20 male, 27 female; age 55±10 years) who both underwent dual-source CTCA and CCA were prospectively included in this study. All patients were performed CTCA using high pitch mode setting at 20%-30% of the R-R interval for the image acquisition. All coronary segments were evaluated by two blinded and independent observers with regard to image quality on a three-point scale (1: excellent to 3: non-diagnostic) and for the presence of significant coronary stenoses (defined as diameter narrowing exceeding 50%). Considered CCA as the standard of reference, the sensitivity, specificity, positive predictive value and negative predictive value were calculated. Radiation dose values were calculated using the dose-length product. Results Image quality was rated as being score 1 in 92.4% of segments, score 2 in 6.1% of segmentsand score 3 in 1.5% of segments. The average image quality score per segment was 1.064±0.306. The HR variability of patients with image score 1, 2 and 3 were 2.29±1.06 bpm, 5.17±1.37 bpm, 8.88±1.53 bpm, respectively. The average HR variability of patients with different image scores were significantly different (F=170.402, P=0.001). The sensitivity, specificity, positive and negative predictive values were 92.6%, 97.0%, 87.6%, 98.3%, respectively, per segment and 90.0%, 95.2%, 85.3%, 96.9%, respectively, per vessel and 100%, 63.6%, 90.0%, 100%, respectively, per patient. The effective radiation dose was on average 0.86±0.16 mSv. Conclusion In patients with HR more than 65 bpm and below 100 bpm without cardiac arrhythmia, the prospectively electrocardiogram-gated high-pitch spiral acquisition mode with image acquired timing set at 20%-30% of the R-R interval provides a high diagnostic accuracy for the assessment of coronary stenoses combined with a 1.5% of non-diagnostic coronary segments and a radiation dose below 1 mSv.

Intravenous Contrast Material Administration at High-pitch Dual-source CT Coronary Angiography: Bolus-tracking Technique with Shortened Time of Respiratory Instruction Versus Test Bolus Technique.

Objective To investigate the feasibility of acquiring the similar homogeneous enhancement using bolus-tracking techniques with shortened respiratory time in prospectively electrocardiogram-gated high-pitch spiral acquisition mode (Flash mode) coronary computed tomography angiography (CCTA) compared with test bolus technique. Methods One hundred and eighty-four consecutive patients with mean heart rate ≤65 beats per minute undergoing CCTA were prospectively included in this study. The patients were randomly divided into two groups. Patients in the group A (n=92) instructed to shorten respiratory time received CCTA using bolus-tracking technique with high-pitch spiral acquisition mode (Flash mode), while those in the group B (n=92) underwent CCTA with test bolus technique. The attenuation in the ascending aorta, image noise, contrast-to-noise ratio and radiation doses of the two groups were assessed. Results There were no significant differences in the mean attenuation values in the ascending aorta (483.18±59.07 HU vs. 498.7±83.51 HU, P=0.183), image noise (21.4±4.5 HU vs. 20.9±4.3 HU, P=0.414), contrast-to-noise ratio (12.1±4.2 vs. 13.8±5.1, P=0.31) between the groups A and B. There were no significant differences in the radiation dose of dynamic monitoring scans (0.056±0.026 mSv vs. 0.062±0.018 mSv, P=0.068) and radiation dose of angiography (0.94±0.07 mSv vs. 0.96±0.15 mSv, P=0.926) between the two groups, while 15 mL less contrast material volume was administered in the group A than the group B. Conclusion Bolus-tracking technique with shortened time of respiratory in Flash mode of dual-source CT yields the similar homogeneous enhancement with less contrast material in comparison to the test bolus technique.

Clinicopathologic features of Stewart-Treves syndrome.

AIMS: To demonstrate clinicopathologic features of Stewart-Treves syndrome (STS) including clinical manifestations, morphology, immunophenotype (especially c-MYC amplification), differential diagnosis, pathogenesis, treatment and prognosis. METHODS AND RESULTS: 17 cases of STS were retrospectively archived, involving 6 cases of postmastectomy, 3 cases of postoperative cervical cancer and 8 cases of chronic lymphatic obstruction without history of malignancy. Seven of 9 cancer patients had undergone radiotherapy. All the patients presented with lymphedema as the first sign. The lesions appeared as multiple reddish blue macules or nodules with polypoid and coalesce. Microscopic examination revealed infiltrative proliferation of irregular vessels in dermis and subcutaneous tissue. The tumorous endothelial cells displayed pleomorphism in morphology. The heteromorphic tumor cells expressed CD34, CD31, ERG, D2-40, c-MYC and factor VIII. Despite various treatment modalities, all cases died in an average of 13.6 months, with 1 case of loss to follow-up. CONCLUSIONS: STS is an extremely rare malignancy that arises from congenital or secondary chronic lymphedema. STS uniquely overexpressed c-MYC. In spite of poor prognosis, early detection is important to facilitate a full range of available therapies, even an opportunity for curative treatment. A low threshold for biopsy and early referral to an experienced multidisciplinary team are highly recommended for optimum management.

Breast carcinoma with micropapillary features: clinicopathologic study and long-term follow-up of 100 cases.

To study the clinicopathologic characteristics and prognosis of invasive micropapillary carcinoma of breast (IMPC), 100 cases of invasive breast carcinoma with an IMPC component were reviewed. Compared with invasive ductal carcinoma, not otherwise specified, with similar histologic grades, carcinomas with IMPC were larger sized, had a higher lymph node metastasis rate with more nodes involved per case, and exhibited increased lymphovascular invasion. The presence of IMPC strongly correlated with the more aggressive behavior. No significant association was established between the proportion of the IMPC component and overall tumor size, histologic grade, lymph node metastasis rate, and distant metastasis, but a trend was noted. Long-term follow-up demonstrated a poorer 5-year and 10-year survival rate for patients with breast carcinoma containing an IMPC component. Breast carcinomas with micropapillary features are more aggressive tumors with a poorer prognosis. This specific structure should be carefully evaluated in the surgical pathology examination of breast carcinoma specimens.

[Significance of expression of stromal cell derived factor 1 and CXCR4 in invasive breast cancer].

OBJECTIVE: To study the expression of stromal cell derived factor 1(SDF-1)/CXCR4 and their association with clinicopathologic features and lymph node metastasis in invasive breast carcinoma. METHODS: The expression of SDF-1 was studied by immunohistochemistry and in-situ hybridization. Immunohistochemical study for CXCR4 was also performed. The correlation with various clinicopathologic parameters was analyzed. RESULTS: (1) SDF-1 was mainly expressed in tumor cells and the level of its expression (both membranous and cytoplasmic) in lymph node-positive group was higher than that in lymph node-negative group (P = 0.033). Only cytoplasmic expression correlated with the number of positive lymph node involved by metastasis, TNM tumor stage, histologic grade, tumor dimension and estrogen receptor status (P < 0.05). (2) SDF-1 protein was also detected in the endothelial cells, although its mRNA was rarely detected. SDF-1 staining in lymphatics was associated with positive lymph node (P = 0.005) and SDF-1 staining in blood vessels correlated with stromal lymphocytic reaction (P = 0.001). The extent of nodal involvement was higher in the group with positive SDF-1 staining in blood vessels and with prominent lymphocytic reaction than that in other groups with one or neither of the two features (P < 0.05). (3) On the other hand, CXCR4 was mainly expressed in tumor cells (both nuclear and cytoplasmic); and the level of its expression in lymph node-positive group was higher than that in lymph node-negative group (P = 0.005). Only cytoplasmic expression correlated with the number of positive lymph node involved by metastasis, TNM tumor stage, histologic grade, tumor dimension and HER2 status (P < 0.05). The nuclear expression of CXCR4 was only correlated with progesterone receptor status (P < 0.01). The cytoplasmic expression CXCR4 also positively correlated with SDF-1 expression (P = 0.001). CONCLUSIONS: SDF-1 and CXCR4 can serve as biomarkers for diagnosis and prediction of lymph node metastasis, as well as potential therapeutic targets in invasive breast carcinoma. The difference in localization and staining patterns may also carry different significance.

[Significance of interleukin-1beta expression and microvascular density in invasive micropapillary carcinoma of breast].

OBJECTIVE: To study the significance of interleukin-1beta (IL-1beta) expression and microvascular density (MVD) in invasive micropapillary carcinoma (IMPC) of breast. METHODS: Immunohistochemical study for IL-1beta and CD34 was performed on 100 cases of IMPC and 97 cases of invasive ductal carcinoma (IDC). The relationship between IL-1beta expression, MVD and various pathologic parameters (estrogen and progesterone receptor status, Ki-67 proliferative index, histologic grade and lymph node metastasis) in IMPC was analyzed. RESULTS: There was no significant difference in expression of IL-1beta between IMPC and IDC (P = 0.924). The expression of IL-1beta positively correlated with proliferative index (P = 0.023), histologic grade (P = 0.038) and lymph node metastasis (P = 0.008), and negatively correlated with estrogen receptor expression (P = 0.035). The MVD in IMPC was significantly higher than that in IDC (66.4 versus 60.0, P = 0.003). The mean MVD in node-positive IMPC was higher than that in node-negative IMPC (68.8 versus 54.4, P = 0.001). In IMPC, the MVD in histologic grade II and III tumors was much higher than that in histologic grade I tumors (68.3 versus 59.9, P = 0.025). It had no relationship with hormonal receptor status and proliferative index. CONCLUSION: Overexpression of IL-1beta and high microvessel density may have important roles in tumor cell proliferation and lymph node metastasis in IMPC.

Responses of eastern Chinese coastal salt marshes to sea-level rise combined with vegetative and sedimentary processes.

The impacts of sea-level rise (SLR) on coastal ecosystems have attracted worldwide attention in relation to global change. In this study, the salt marsh model for the Yangtze Estuary (SMM-YE, developed in China) and the Sea Level Affecting Marshes Model (SLAMM, developed in the U.S.) were used to simulate the effects of SLR on the coastal salt marshes in eastern China. The changes in the dominant species in the plant community were also considered. Predictions based on the SLAMM indicated a trend of habitat degradation up to 2100; total salt marsh habitat area continued to decline (4-16%) based on the low-level scenario, with greater losses (6-25%) predicted under the high-level scenario. The SMM-YE showed that the salt marshes could be resilient to threats of SLR through the processes of accretion of mudflats, vegetation expansion and sediment trapping by plants. This model predicted that salt marsh areas increased (3-6%) under the low-level scenario. The decrease in the total habitat area with the SMM-YE under the high-level scenario was much lower than the SLAMM prediction. Nevertheless, SLR might negatively affect the salt marsh species that are not adapted to prolonged inundation. An adaptive strategy for responding to changes in sediment resources is necessary in the Yangtze Estuary.

[Vulnerability assessment on the coastal wetlands in the Yangtze Estuary under sea-level rise].

To study the response of coastal wetlands to climate change, assess the impacts of climate change on the coastal wetlands and formulate feasible and practical mitigation strategies are the important prerequisite for securing coastal ecosystems. In this paper, the possible impacts of sea level rise caused by climate change on the coastal wetlands in the Yangtze Estuary were analyzed by the Source-Pathway-Receptor-Consequence (SPRC) model and IPCC definition on the vulnerability. An indicator system for vulnerability assessment was established, in which sea-level rise rate, subsidence rate, habitat elevation, inundation threshold of habitat and sedimentation rate were selected as the key indicators. A quantitatively spatial assessment method based on the GIS platform was established by quantifying each indicator, calculating the vulnerability index and grading the vulnerability index for the assessment of coastal wetlands in the Yangtze Estuary under the scenarios of sea-level rise. The vulnerability assessments on the coastal wetlands in the Yangtze Estuary in 2030 and 2050 were performed under two sea-level rise scenarios (the present sea-level rise trend over recent 30 years and IPCC A1F1 scenario). The results showed that with the projection in 2030 under the present trend of sea-level rise (0.26 cm x a(-1)), 6.6% and 0.1% of the coastal wetlands were in the low and moderate vulnerabilities, respectively; and in 2050, 9.8% and 0.2% of the coastal wetlands were in low and moderate vulnerabilities, respectively. With the projection in 2030 under the A1F1 scenario (0.59 cm x a(-1)), 9.0% and 0.1% of the coastal wetlands were in the low and moderate vulnerabilities, respectively; and in 2050, 9.5%, 1.0% and 0.3% of the coastal wetlands were in the low, moderate and high vulnerabilities, respectively.

Gene cloning, expression, and substrate specificity of an imidase from the strain Pseudomonas putida YZ-26.

A gene-encoding imidase was isolated from Pseudomonas putdia YZ-26 genomic DNA using a combination of polymerase chain reaction and activity screening the recombinant. Analysis of the nucleotide sequence revealed that an open reading frame (ORF) of 879 bp encoded a protein of 293 amino acids with a calculated molecular weight of 33712.6 kDa. The deduced amino-acid sequence showed 78% identity with the imidase from Alcaligenes eutrophus 112R4 and 80% identity with N-terminal 20 amino-acid imidase from Blastobacter sp. A17p-4. Next, the ORF was subcloned into vector pET32a to form recombinant plasmid pEI. The enzyme was overexpressed in Escherichia coli and purified to homogeneity by Ni(2+)-NTA column, with 75% activity recovery. The subunit molecular mass of the recombinant imidase as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was approximately 36 kDa, whereas its functional unit was approximately 141 kDa with four identical subunits determined by size-exclusion chromatography. The purified enzyme showed the highest activity and affinity toward succinimide, and some other substrates, such as dihydrouracil, hydantoin, succinimide, and maleimde, were investigated.