A Gene Transfer-Positive Cell Sorting System Utilizing Membrane-Anchoring Affinity Tag.

Gene delivery efficiency is an essential limit factor in gene study and gene therapy, especially for cells that are hard for gene transfer. Here we develop an affinity cell sorting system that allows efficient enrichment of gene transfer-positive cells. The system expresses an enhanced green fluorescent protein (EGFP) fused with an N-terminal high-affinity Twin-Strep-Tag (TST) that will be anchored to the cell membrane at the out-surface through a glycosylphosphatidylinositol (GPI) membrane-anchoring structure. The EGFP permits microscopy and flow cytometry analysis of the gene transfer-positive cells, and the TST tag at the N terminal of EGFP allows efficient affinity sorting of the positive cells using Strep-Tactin magnetic beads. The cell sorting system enables efficient isolation of gene transfer-positive cells in a simple, convenient, and fast manner. Cell sorting on transfected K-562 cells resulted in a final positive cell percentage of up to 95.0% with a positive cell enrichment fold of 5.8 times. The applications in gene overexpression experiments could dramatically increase the gene overexpression fold from 10 times to 58 times, and in shRNA gene knockdown experiments, cell sorting increased the gene knockdown efficiency from 12% to 53%. In addition, cell sorting in CRISPR/Cas9 genome editing experiments allowed more significant gene modification, with an editing percentage increasing from 20% to 79%. The gene transfer-positive cell sorting system holds great potential for all gene transfer studies, especially on those hard-to-transfect cells.

Function of Atg11 in non-selective autophagy and selective autophagy of Candida albicans.

Candida albicans is an important opportunistic pathogenic fungus in the human body. It is a common microbe inhabiting on the mucosa surfaces of healthy individuals, but may cause infections when the host immune system is weak. Autophagy is a "self-eating" process in eukaryotes, which can recover and utilize damaged organelles and misfolded proteins. Here we investigated the role of the autophagy-related protein Atg11 in C. albicans. Deletion of ATG11 led to the defect in growth under the nitrogen starvation condition. Western blotting and GFP localization further revealed that the transport and degradation of Atg8 was blocked in the atg11Δ/Δ mutant under both the nitrogen starvation and hypha-inducing conditions. Moreover, degradation of both Lap41 (the indicator of the cytoplasm-to-vacuole pathway) and Csp37 (the indicator of mitophagy) was also thoroughly suppressed in this mutant under nitrogen starvation. These results indicated that Atg11 plays an essential role in both non-selective and selective autophagy in C. albicans.

Association study of cell cycle proteins and human papillomavirus in laryngeal cancer in Chinese population.

OBJECTIVE: To investigate the role of human papillomavirus (HPV) in laryngeal squamous cell carcinoma (LSCC) and analyse the relationship between HPV and the expression of cell cycle-related proteins. DESIGN: The study consisted of LSCC between 2005 and 2011 in Tongren Hospital. Clinical data such as age, sex, smoking/alcohol consumption, and TNM stage were collected. HPV DNA and cell cycle-related proteins were assessed in terms of clinical features. SETTING: Single-centre study. PARTICIPANTS: A total of 332 LSCC patients were included in the study. MAIN OUTCOME MEASURES: The presence of genotype-specific HPV DNA was evaluated using PCR-RDB in formalin-fixed paraffin-embedded tissues. All samples were also evaluated for p16INK4A , p21WAF1/CIP1 , P53, Cyclin D1, and Ki67 immunohistochemical staining by tissue microarray. RESULTS: HPV DNA was detected in 45 of 332 (13.55%) patients with LSCC, with HPV-16 being the predominant genotype. The presence of HPV-16 DNA was significantly associated with basaloid squamous cell carcinoma and cystic lymph node metastasis (P < 0.05). Of the 332 patients, 36 (10.84%) were scored as p16INK4A positivity and they were more likely to be female (P < 0.05). Cyclin D1-positivity and p21WAF1/CIP1 -positivity were observed in 60.24% (200/332) and 40.66% (135/332), respectively. In 114 cases (34.33%), LSCCs had moderate- to -strong p53 accumulation, which was correlated with TNM stage (P < 0.05). HPV-16 DNA was correlated with p16INK4A and manifested a higher Ki-67 labelling index and p21WAF1/CIP1 expression than HPV-16-negative tumours (P < 0.05). No relationship was observed between Cyclin D1or P53 expression and HPV-16 infection (P > 0.05). CONCLUSION: HPV DNA was detected in 13.55% patients with LSCC, with HPV-16 being the predominant genotype and it was correlated with p16INK4A and manifested a higher Ki-67 labelling index and p21WAF1/CIP1 expression than HPV-16-negative tumours.

Erbin plays a critical role in human umbilical vein endothelial cell migration and tubular structure formation via the Smad1/5 pathway.

Angiogenesis is an important process in atherosclerosis. ErbB2 was proved to have an important role in vascular development, but it is still unclear whether Erbin expresses in vessels as well as its location and function in the vessels. In the current study, we investigated the location and function of Erbin in human umbilical veins. The human umbilical veins were prepared, and immunofluorescent analysis was performed to determine the expression of Erbin. Human umbilical vein endothelial cells (HUVECs) were cultured and the lentivirus (LV) containing Erbin RNAi was also prepared. After transfection with the lentivirus, CCK-8 assay and Annexin V-PI assay were used for cell proliferation and apoptosis, respectively. Cell migration was studied using the scratch wound healing assay and the transwell assay. The capillary-like tube formation assay was performed to illustrate the effect of Erbin on HUVEC tube formation. Expression of signaling pathway molecules was assessed with Western blot. The immunofluorescent analysis suggested that Erbin expressed in human umbilical veins and the majority of the Erbin is strongly colocalized in endothelial cells. Although knockdown of Erbin did not affect HUVEC proliferation and apoptosis, it significantly suppressed HUVEC migration and tubular structure formation. Erbin knockdown showed no effect on the ERK1/2 and Smad2/3 signaling pathways but significantly promoted Smad1/5 phosphorylation and nuclear translocation. Ablation of the Smad1/5 pathway decreased the effects of Erbin on endothelial cells. Erbin is mainly localized in endothelial cells in human umbilical veins and plays a critical role in endothelial cell migration and tubular formation via the Smad1/5 pathway.

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.

Mgm1 is required for maintenance of mitochondrial function and virulence in Candida albicans.

Mitochondria are dynamic organelles, and their shapes and sizes are regulated by mitochondrial fusion and fission. The proteins essential for mitochondrial fusion in Candida albicans have not been clearly characterized. In this study, Mgm1 was explored for its roles in mitochondrial function, cell cycle, hyphal growth and virulence in this pathogen. The deletion of MGM1 led to mitochondrial fragmentation and mtDNA loss and activated the checkpoint pathway to arrest the cell cycle in G1 phase. Moreover, loss of MGM1 led to defects in hyphal development and attenuation of virulence in a macrophage cell line and a mouse model of disseminated infection. These results reveal that Mgm1 plays an important role in mitochondrial dynamics and function, cell cycle progression, hyphal development and virulence in C. albicans.

Association analysis between spatiotemporal variation of vegetation greenness and precipitation/temperature in the Yangtze River Basin (China).

The variation in vegetation greenness provides good understanding of the sustainable management and monitoring of land surface ecosystems. The present paper discusses the spatial-temporal changes in vegetation and controlling factors in the Yangtze River Basin (YRB) using Global Inventory Modeling and Mapping Studies (GIMMS) Normalized Difference Vegetation Index (NDVI) for the period 2001-2013. Theil-Sen Median trend analysis, Pearson correlation coefficients, and residual analysis have been used, which shows decreasing trend of the annual mean NDVI over the whole YRB. Spatially, the regions with significant decreasing trends were mainly located in parts of central YRB, and pronounced increasing trends were observed in parts of the eastern and western YRB. The mean NDVI during spring and summer seasons increased, while it decreased during autumn and winter seasons. The seasonal mean NDVI shows spatial heterogeneity due to the vegetation types. The correlation analysis shows a positive relation between NDVI and temperature over most of the YRB, whereas NDVI and precipitation show a negative correlation. The residual analysis shows an increase in NDVI in parts of eastern and western YRB and the decrease in NDVI in the small part of Yangtze River Delta (YRD) and the mid-western YRB due to human activities. In general, climate factors were the principal drivers of NDVI variation in YRB in recent years.

Human papillomavirus infection and its biomarkers' expressions in laryngeal basaloid squamous cell carcinoma.

AIMS: To investigate the frequency and transcriptional activity of HPV and its correlation to p16 and p21 expression in basaloid squamous cell carcinoma (BSCC) of the larynx. METHODS: We evaluated tissues from 29 patients with BSCC of the larynx for the expressions of p16 and p21 proteins by immunohistochemistry (IHC) and for HPV E6 and E7 mRNA by RNA in situ hybridization (ISH). The presence of genotype-specific HPV DNA was evaluated using PCR-RDB in formalin-fixed paraffin-embedded tissues. P16 and p21 expression and HPV DNA status were correlated with clinicopathological features. RESULTS: HPV DNA was detected in 8 of 29 (27.59%) patients, with HPV-16 being the predominant genotype. P16 and p21-positivity were observed in 7/29 (24.14%) and 8/29 (27.59%) patients, respectively. HPV was not correlated with p16 expression (P > 0.05). However, p21 expression was significantly higher in HPV-positive tumors than in HPV-negative tumors (P < 0.05). No cases exhibited transcriptionally active HPV in our series. CONCLUSION: Our findings suggest that a small fraction of BSCC of the larynx is HPV DNA-positive in this Chinese population, p21 expression was significantly higher in HPV-positive tumors, and no cases were HPV transcriptionally active in this small cohort. Further research of HPV and its role in BSCC of the larynx are warranted.

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.

Angiosarcoma (Stewart-Treves syndrome) in postmastectomy patients: report of 10 cases and review of literature.

AIMS: To study the clinicopathologic features of Stewart-Treves syndrome (STS) in postmastectomy patients including the epidemiology, presentation, morphology, differentiation, pathogenesis and therapeutic options. METHODS AND RESULTS: Ten cases of STS in postmastectomy patients were retrospectively identified in our archives, and immunohistochemistry for CD34, CD31, D2-40, HHV-8, CK, EMA and Ki-67 was performed. All ten patients presented with lymphedema after mastectomy as the first sign. Physical examination revealed multiple raised, pinkish-red papulo-vesicular lesions or ulceration as the early evidence of tumor in the field where radiation therapy was introduced. Microscopic examination revealed infiltrative proliferation of vessels and the heteromorphic tumor cells expressed CD34, CD31 and D2-40. Despite the various treatment modalities, 5 patients died in an average of 19 months, 4 patients survived to the last follow-up (9-31 months), and 1 patient got lost. CONCLUSIONS: STS is a fatal complication of postmastectomy lymphedema. Patients with STS have very poor prognosis. The key to improve patient's survival is the early diagnosis through a high alert of this disease by primary care physicians and comprehensive physical examination of patients with pertinent history and suspicious clinical presentations followed by prompt biopsy for definitive diagnosis.

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.

Polypoid colonic metastases from gastric stump carcinoma: A case report.

The present study aimed to investigate polypoid colonic metastases from gastric stump carcinoma by performing a retrospective analysis of the clinical data of a patient with such a diagnosis, and by discussing other previous case studies from the literature. The patient of the present study was an 80-year-old male who had undergone a gastrectomy 48 years previously for a benign perforated gastric ulcer. A colonoscopy revealed >10 multiple polypoid lesions of 6-10 mm in diameter distributed throughout the entire colon, except in the rectum. Each lesion had either erosion or a depression at the top and several were covered with a white fur-like substance. Biopsy specimens excised from the stomach showed a poorly-differentiated adenocarcinoma with diffuse signet ring cells, and a colonoscopy-guided biopsy revealed a signet ring cell adenocarcinoma. The patient was referred to the Oncology unit (Beijing Shijitan Hospital, Beijing, China) for assessment and chemotherapy treatment, which was initiated with 1,000 mg Xeloda orally administered twice a day for two-week courses every three weeks. The patient succumbed to upper gastrointestinal hemorrhage and pneumonia after three months. Gastric or gastric stump carcinoma may metastasize to the colon presenting as solitary or multiple colonic polyps. Thus, it is important to consider this diagnosis as such colon metastases may mimic solitary or multiple colonic polyps, which are commonly observed. A differential diagnosis is required in this complicated situation.

Rabbit antiserum to mouse embryonic stem cells delays compaction of mouse preimplantation embryos.

BACKGROUND: Mouse embryonic stem (ES) cells are derived from the inner cell mass (ICM) of the preimplantation blastocysts. So it is suggested that ES and ICM cells should have similar cellular surface molecules and antiserum to ES cells can inhibit ICM development. OBJECTIVE: The objective of this study was to evaluate the effect of rabbit antiserum to ES cells on mouse preimplantation embryo development and chimera production. MATERIALS AND METHODS: Mouse 4-cell embryos were matured in vitro at 37.5(o)C, in humidified 5% CO2 atmosphere for 12-36 h. The embryos were cultured in KSOM medium with or without antiserum for 12-36 h. The ratios of in vitro embryo development of the blastocysts, cell division, attachment potential, alkaline phosphatase activity, post-implantation development, and chimera production were assessed and compared with the control group. P<0.05 was considered as significant. RESULTS: The rabbit antiserum to mouse ES cells showed delay in embryo compaction and induced decompaction at 8-cell stage. The development of 4-cell embryos in the presence of the antiserum for 36h did not lead to a reduced or absent ICM. These embryos still displayed positive alkaline phosphatase activity, normal cell division, embryo attachment, outgrowth formation, implantation and post-implantation development. In addition, decompaction induced by antiserum did not increase production and germline transmission of chimeric mice. CONCLUSION: The results showed that antiserum to ES cells delayed embryo compaction and did not affect post-implantation development and chimera production.

[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.

Transcervical embryo transfer in mice.

Although efficient transcervical transfer of embryos in mice would provide many advantages over a surgical method, the low success rate of transcervical transfer has hampered its acceptance and use. Here, we describe a novel device and protocol for transcervical embryo transfer in mice. Blastocysts from CD1 female mice were transferred into the uteri of 2.5-d pseudopregnant CD1 mice by using this method resulted in the successful development of 66.7% to 73.5% of the transferred blastocysts into live-born fetuses. Our method is as efficient as surgical embryo transfer yet is much simpler, easier, and markedly less traumatic to the recipient. In addition, our method provides hygienic and economic advantages and conforms to the principles of humane experimental technique. More importantly, our method provides a model for studying transcervical embryo transfer in cattle, other large animals, and humans.

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.

Tumor-to-tumor metastasis from pituitary carcinoma to radiation-induced meningioma.

Radiation-induced meningioma and pituitary carcinoma are both uncommon. Tumor-to-tumor metastasis (TTM) from pituitary carcinoma to meningioma, to our knowledge, has not been previously reported. A 67-year old man presented with a previous history of transcranial subtotal resection of pituitary adenoma, at the age of 36, followed by radiotherapy. The follow-up was uneventful for the following 31 years. The patient presented with worsening sight and numbness of the right arm. Three separate lesions were found on MRI. Histological examinations revealed pituitary carcinomas and TTM from pituitary carcinoma to meningioma. A constant surveillance is necessary for patients with pituitary tumor, especially those followed by radiotherapy.

The clinicopathological significance of CD44+/CD24-/low and CD24+ tumor cells in invasive micropapillary carcinoma of the breast.

Breast cancer cells with a CD44(+)/CD24(-/low) phenotype have been suggested to have tumor-initiating properties. It is unclear whether their presence correlates with clinicopathological features of invasive micropapillary carcinoma (IMPC) of the breast, an unusual subtype of breast cancer with a high incidence of lymph node metastasis and poor prognosis. CD44 and CD24 expression was determined by double-staining immunohistochemistry in 103 cases of IMPC and in 94 cases of invasive ductal carcinoma (IDC). The prevalence of CD44(+)/CD24(-/low) tumor cells was higher in IMPC than in invasive ductal carcinoma IDC (P=0.018). The CD44(+)/CD24(-/low) tumor cells were also detected in adjacent stroma surrounding the micropapillary structure in 53.4% (55/103) of IMPC, but only in 7.4% (7/94) of stroma of IDC. These tumor cells in stroma of IMPC were positive for vimentin and α-smooth muscle actin, and negative for E-cadherin. The CD44(+)/CD24(-/low) tumor cells in the micropapillary structure of IMPC were associated with those in stroma (P=0.000). Moreover, they were both associated with lymphovascular invasion and extranodal extension, respectively (P<0.05). The proportion of CD24(+) tumor cells was also higher in IMPC than in IDC (P=0.035), and the CD24(+) tumor cells were associated with lymph node metastasis in IMPC (P=0.010). The results suggest that the increased proportion of CD44(+)/CD24(-/low) tumor cells and CD24(+) tumor cells and the epithelial mesenchymal transition may play an important role in aggressiveness and high metastatic risk of breast IMPC.

Expression of HAb18G is associated with tumor progression and prognosis of breast carcinoma.

HAb18G is a recently identified hepatoma-associated antigen and its association with tumor growth, invasion, and angiogenesis has been studied in a variety of tumors. However, its role in the tumor progression of breast cancer has not been explored. HAb18G expression was examined by immunohistochemistry in pathological sections of 1,637 breast tissue samples and by in situ hybridization in 41 cases of invasive breast carcinomas (BC). While not detected in any cases of tumor-like conditions or benign tumors of breast, and only rarely in normal tissue (4.4%), HAb18G expression was gradually up-regulated from atypical ductal hyperplasia (27.3%), to ductal carcinoma-in situ (59.8%), and to BC (61.4%) (P < 0.01). Its expression in BC was correlated positively with C-erbB-2 expression and histologic grade (P < 0.001), and negatively with the expression of estrogen and progesterone receptors (P < 0.001). Significant differences of expression were also identified among the subgroups of BC examined: in decreasing order from invasive micropapillary carcinoma, ductal carcinoma, lobular carcinoma, papillary carcinoma, medullary carcinoma, to mucinous adenocarcinoma (P = 0.001), corresponding to their known clinical aggressiveness. In an expanded group of 186 BC patients with proper follow up, our previous findings were confirmed: HAb18G expression was significantly associated with local recurrence, distant metastasis and tumor mortality (P < 0.01). We also demonstrated that up-regulated tumor expression of HAb18G was a significant predictor of reduced disease progression-free survival rate and a shorter overall survival, independent of systemic therapies. In conclusion, this study suggests that HAb18G expression is associated with BC progression and prognosis. Further evaluation of this new marker in breast cancer is indicated.