[Relationship between microsatellite instability and hepatocyte growth factor expression and their prognostic significance in colorectal cancer].

OBJECTIVE: To investigate the expression of hepatocyte growth factor (HGF) and its relationship with microsatellite instability (MSI) and their influence on survival in patients with colorectal cancer. METHODS: Immunohistochemistry (IHC) was used to detect the expression of HGF and MSI in 98 specimens of colorectal cancer. Tumors lacking protein expression of any of the four mismatch repair genes (MLH1, PMS2, MSH2 or MSH6) were labelled as MSI, and the rest were considered as microsatellite stable (MSS). The associations between expression and clinicopathological factors were assessed using Chi-square tests. Kaplan-Meier curves, log-rank test, and Cox regression were used to analyze the association between biomarker expressions and overall survival. RESULTS: The incidence rate of MSI in 98 colorectal specimens was 32.7%, and was statistically significantly correlated with the location of tumor and differentiation degree (P<0.05). The HGF-expression rate was 71.4%. The patients with an MSI tumor had a significantly higher HGF expression, compared with the patients with an MSS tumor (P=0.048). The 5-year survival rate of MSI group and MSS group were 39.8% and 58.7%, respectively (P=0.009). The 5-year survival rate of HGF-positive group and HGF-negative group were 46.2% and 67.9% (P=0.035). The multivariate analysis showed that lymphocytic infiltration, TMN stage, MSI and HGF are independent prognostic factors in colorectal cancer (P<0.05 for all). CONCLUSIONS: HGF is highly expressed in colorectal cancer patients with microsatellite instability. Both microsatellite instability and HGF are independent factors affecting the prognosis in patient with colorectal cancer.

Molecular cloning and expression profile of an ATP-binding cassette (ABC) transporter gene from the hemipteran insect Nilaparvata lugens.

The ATP-binding cassette (ABC) transporters belong to a large superfamily of proteins that have important physiological functions in all living organisms. In insects, ABC transporters have important functions in the transport of molecules, and are also involved in insecticide resistance, metabolism, and development. In this study, the Nilaparvata lugens Stal (Hemiptera: Delphacidae) ABCG (NlABCG) gene was identified and characterized. The complete mRNA sequence of NlABCG was 2608-bp long, with an open reading frame of 2064 bp encoding a protein comprised of 687 amino acids. The conserved regions include three N-glycosylation and 34 phosphorylation sites, as well as seven transmembrane domains. The amino acid identity with the closely related species Acyrthosiphon pisum was 42.8%. Developmental expression analysis using quantitative real-time reverse transcriptase PCR suggested that the NlABCG transcript was expressed at all developmental stages of N. lugens. The lowest expression of NlABCG was in the 1st instar, and levels increased with larval growth. The transcript profiles of NlABCG were analyzed in various tissues from a 5th instar nymph, and the highest expression was observed in the midgut. These results suggest that the sequence, characteristics, and expression of NlABCG are highly conserved, and basic information is provided for its functional analysis.

Can abundant colloid exclude oncocytic (Hürthle cell) carcinoma in thyroid fine needle aspiration? Cytohistological correlation of 127 oncocytic (Hürthle cell) lesions.

OBJECTIVE: The objective of the present study was to find out whether the presence or absence of certain cytological features can exclude oncocytic (Hürthle cell) carcinoma in thyroid fine needle aspiration (FNA) to minimize unnecessary surgery. METHODS: Over a 17-year period, 127 hypercellular, oncocyte-exclusive, lymphocyte-absent aspirates obtained via ultrasound-guided FNA with on-site assessment had histology slides for review. The presence or absence of six cytological features (microfollicular arrangement, discohesive single cells, small cell dysplasia, large cell dysplasia, transgressing blood vessels and colloid) and one histological feature (macrofollicular component) were determined for each case independently by two cytopathologists. RESULTS: Histology showed 12 (9.4%) cases of Hashimoto thyroiditis, 23 (18.1%) oncocytic adenomatoid nodules in nodular goitre, 66 (52.0%) oncocytic adenomas and 26 (20.5%) oncocytic carcinomas (13 minimally invasive without angioinvasion, six minimally invasive with angioinvasion, seven widely invasive). Histologically, a macrofollicular component was present in seven of 26 (26.9%) oncocytic carcinomas, including one case with abundant thin colloid. A microfollicular arrangement, discohesive single cells, small cell dysplasia, large cell dysplasia and transgressing vessels were present in oncocytic carcinoma, oncocytic adenoma and oncocytic adenomatoid nodules in nodular goitre. CONCLUSIONS: A macrofollicular component is frequently present in oncocytic carcinoma, oncocytic adenoma and oncocytic adenomatoid nodules in nodular goitre. None of the cytological features studied, including abundant colloid, can exclude oncocytic carcinoma. Oncocytic carcinoma can only be excluded by thorough histological examination of thyroidectomy specimens. A molecular marker is needed to triage oncocytic lesions in thyroid FNA.

Detection of medullary thyroid microcarcinoma using ultrasound-guided fine needle aspiration cytology.

OBJECTIVE: Compared with incidental papillary thyroid microcarcinoma (microPTC), incidental medullary thyroid microcarcinoma (microMTC) is clinically more significant. The objective of the present study was to summarize our experience in detecting microMTCs. METHODS: From 1995 to 2011, there were 10825 thyroid fine needle aspirates (FNAs) guided using high-resolution ultrasound with on-site preparation and evaluation by a cytopathologist. Of the 140 microcarcinomas detected, 132 were microPTCs and eight were microMTCs, which are the subject of the present study. RESULTS: All eight cases were incidentalomas and none of the five women and three men, age 37-70 years, had a family history of MTC. One patient had two FNAs at an interval of 10 months, two had a single lymph node metastasis and one had a 0.1-cm tumour nodule near the main tumour. Four of five plasmacytoid cell microMCTs had irregular borders; two round cell and one rectangular cell tumours had smooth borders. In contrast, 17 larger MTCs diagnosed in the same period included seven plasmacytoid, four giant cell and six spindle cell types. All five plasmacytoid microMTCs were correctly diagnosed on FNA, but the round cell and rectangular cell tumours were undercalled as follicular lesions. Sampling of colloid from adjacent follicles was noted in microMTCs. Two were diagnosed on histology following recommended surgery and one was diagnosed on recommended repeat FNA. CONCLUSIONS: US-guided FNA of thyroid lesions is a powerful tool in the detection of microMTCs, provided that cytopathologists are alerted to the pitfalls described in the present study.

Mapping and fine mapping of quantitative trait loci for the number of vertebrae in a White Duroc × Chinese Erhualian intercross resource population.

The number of vertebrae is associated with body size and meat production in pigs. To identify quantitative trait loci (QTL) for the number of vertebrae, phenotypic values were measured in 1029 individuals from a White Duroc × Chinese Erhualian intercross F(2) population. A whole genome scan was performed with 194 microsatellite markers in the F(2) population. Four genome-wide significant QTL and eight chromosome-wide significant QTL for the number of vertebrae were identified on pig chromosomes (SSC) 1, 2, 6, 7, 10 and 12. The most significant QTL was detected on SSC7 with a confidence interval of 1 cM, explaining 42.32% of the phenotypic variance in the thoracic vertebral number. The significant QTL on SSC1, 2 and 7 confirmed previous reports. A panel of 276 animals representing seven Western and Chinese breeds was genotyped with 34 microsatellite markers in the SSC7 QTL region. No obvious selective sweep effect was observed in the tested breeds, indicating that intensive selection for enlarged body size in Western commercial breeds did not wipe out the genetic variability in the QTL region. The Q alleles for increased vertebral number originated from both Chinese Erhualian and White Duroc founder animals. A haplotype block of approximately 900 kb was found to be shared by all Q-bearing chromosomes of F(1) sires except for one distinct Q chromosome. The critical region harbours the newly reported VRTN gene associated with vertebral number. Further investigations are required to confirm whether VRTN or two other positional candidate genes, PROX2 and FOS, cause the QTL effect.

The modulation of electronic and optical properties of OXD-X through introduction of the electron-withdrawing groups: a DFT study.

A comparative study using density functional theory (DFT) on the molecular structure, electronic structure and relative properties of 1,3,4-oxadiazole dimers 1,3-bis[2-(4-methylphenyl)-1,3,4-oxadiazol-5-yl]benzene (OXD-X) and its derivatives with alkoxy substituents -O(CH(2))(n-1)CH(3) (OXD-An, n=1, 2) and electron-withdrawing substituents -CN (OXD-C), -CF(3) (OXD-TFM), -NO(2) (OXD-N) added at meta-substitution in the phenyl ring are presented. The ground state structures of the title complexes are optimized at B3LYP/6-31G level of theory. In addition, the time dependent density functional theory (TDDFT) method is applied to calculate the absorption and emission spectra of the derivatives based on the ground state geometries. Comparing with the alkoxy substituents, the results show that the electron-withdrawing substituents have remarkable influences on the energy levels of the frontier molecular orbitals, the spectra and the transition compositions. Especially, -NO(2) group plays the prominent role and the fluorine improves the energy level of LUMO further. The experimental absorption wavelengths for OXD-X, OXD-A3 and OXD-A7 are well reproduced by the TDDFT technique. Moreover the absorption wavelengths and the transition compositions can be effectively adjusted through introducing electro-withdrawing groups in the phenyl ring. The reorganization energies for hole and electron are smaller than that of typical hole and electron transport materials.

A genome scan for quantitative trait loci affecting male reproductive traits in a White Duroc x Chinese Erhualian resource population.

Chinese Erhualian boars have dramatically smaller testes, greater concentrations of circulating androgens, and fewer Sertoli cells than Western commercial breeds. To identify QTL for boar reproductive traits, testicular weight, epididymal weight, seminiferous tubular diameter at 90 and 300 d, and serum testosterone concentration at 300 d were measured in 347 F(2) boars from a White Duroc x Chinese Erhualian cross. A whole genome scan was performed with 183 microsatellites covering 19 porcine chromosomes. A total of 16 QTL were identified on 9 chromosomes, including 1% genome-wide significant QTL for testicular weight at 90 and 300 d and seminiferous tubular diameter at 90 d on SSCX, and for epididymal weight and testosterone concentration at 300 d on SSC7. Two 5% genome-wide significant QTL were detected for testicular weight at 300 d on SSC1 and seminiferous tubular diameter at 300 d on SSC16. Nine suggestive QTL were found on SSC1, 2, 3, 5, 7, 13, and 14. Chinese Erhualian alleles were not systematically favorable for greater reproductive performance. This study confirmed the previous significant QTL for testicular weight on SSCX and for epididymal weight on SSC7, and reported QTL for seminiferous tubular diameter and testosterone concentration at the first time. The observed different QTL for the same trait at different ages reflect the involvement of distinct genes in the development of male reproductive traits.

Treatment of nanosized TiO2-containing wastewater by simultaneous electrocoagulation/electrofiltration.

In this work, a simultaneous electrocoagulation/electrofiltration (EC/EF) treatment module was employed to treat nanosized TiO2-containing wastewater. Nanosized TiO2-containing wastewater was obtained and treated by a self-designed EC/EF treatment module. To evaluate the performance of this novel treatment module, the effects of electric field strength (EFS), transmembrane pressure (TMP), and crossflow velocity (CV) on permeate qualities were investigated. Permeate qualities of concern included pH, turbidity, conductivity, chemical oxygen demand (COD), and total organic carbon (TOC). A full factorial design of experiments was adopted in this work. First, by keeping TMP and CV constant the effects of EFS on permeate qualities were studied. In this set of testing, it was noticed that an application of electric field greatly increased the filtration rate, which was further influenced by the magnitude of EFS. In all cases, the filtration rate decreased as the treatment time elapsed due mainly to fouling of the membrane. Further tests were conducted to study the effects of TMP on permeate qualities by keeping EFS and CV constant. Finally, the effects of CV on permeate qualities were studied by keeping EFS and TMP constant. It was found that the optimal operating conditions would be electric field strength of 166.7 V/cm, transmembrane pressure of 1 kgf/cm2, and crossflow velocity of 0.22cm/s. Under such conditions, permeate would have the following qualities: (1) pH, 6.32; (2) turbidity, 2.41 NTU; (3) conductivity, 15.11 microS/cm; (4) COD, 100.0 mg/L; and (5) TOC, 512.6 mg/L.

A preliminary study on electrically enhanced crossflow microfiltration of CMP (chemical-mechanical polishing) wastewater.

Chemical-mechanical polishing (CMP) is currently one of the main technologies used by wafer plants in the semiconductor industry. Normally, a large amount of de-ionized water should be used to wash out the abrasives adhered to the surface of wafers during the grinding process. Therefore, CMP wastewater not only has a great quantity but also contains very small size of suspended solids. Generally, these suspended solids would not settle. This phenomenon results in a low visibility of CMP wastewater. To solve this problem, a study on electrically enhanced crossflow microfiltration of CMP wastewater was conducted. Normally, the membrane of traditional dead-end filtration is easily blocked by the filter cake resulting in a small flux of filtrate. Therefore, the form of crossflow filtration (CFF) is used to reduce the blocking of the membrane. Furthermore, if CFF is accompanied by an external electric field, the negatively charged suspended solids in wastewater would move toward the positive electrode. Meanwhile, the flux of filtrate would increase. In this investigation, CMP wastewater was obtained from a wafer plant and characterized by various standard methods. Before testing, the CMP wastewater was pre-filtered using a 1.2 microm pore size filter. Then it was operated by a crossflow microfiltration (0.1 microm pore size) system under an appropriate crossflow velocity, filtration pressure, and electric potential. The filtrate was also characterized by various standard methods. Experimental results show that the filtrate has a turbidity of zero value. Thus, the filtrate could be reused for other purposes. However, the suspended solids of the CMP wastewater would be concentrated by this method. Therefore, the recovery of the suspended solids would be worth considering.

Remediation of TCE contaminated soils by in situ EK-Fenton process.

The treatment performance and cost analysis of in situ electrokinetic (EK)-Fenton process for oxidation of trichloroethylene (TCE) in soils were evaluated in this work. In all experiments, an electric gradient of 1V/cm, de-ionized water as the cathode reservoir fluid and a treatment time of 10 days were employed. Treatment efficiencies of TCE were evaluated in terms of the electrode material, soil type, catalyst type, and catalyst dosage and granular size if applicable. Test results show that graphite electrodes are superior to stainless steel electrodes. It was found that the soil with a higher content of organic matter would result in a lower treatment efficiency (e.g. a sandy loam is less efficient than a loamy sand). Experimental results show that the type of catalyst and its dosage would markedly affect the reaction mechanisms (i.e. "destruction" and "removal") and the treatment efficiency. Aside from FeSO4, scrap iron powder (SIP) in the form of a permeable reactive wall was also found to be an effective catalyst for Fenton reaction to oxidize TCE. In general, the smaller the granular size of SIP, the lower the overall treatment efficiency and the greater the destruction efficiency. When a greater quantity of SIP was used, a decrease of the overall treatment efficiency and an increase of percent destruction of TCE were found. Experimental results have shown that the quantity of electro-osmotic (EO) flow decreased as the quantity of SIP increased. It has been verified that the treatment performances are closely related to the corresponding EO permeability. Results of the cost analysis have indicated that the EK-Fenton process employed in this work is very cost-effective with respect to TCE destruction.

Ultrasound-guided fine-needle aspiration of the thyroid assessed by Ultrafast Papanicolaou stain: data from 1135 biopsies with a two- to six-year follow-up.

One of the limitations of fine-needle aspiration (FNA) of the thyroid is difficulty in distinguishing the follicular variant (FV) of papillary thyroid carcinomas (PTC) from follicular neoplasms. By highlighting the "Orphan Annie-eyed" clear nuclei of the former, the Ultrafast Papanicolaou stain (UFP) easily separates these two entities. One thousand one hundred thirty-five ultrasound-guided FNAs of the thyroid were assessed by UFP with immediate biopsy results reported to the patients in a busy radiology office in Manhattan from November 1994 to December 1998. Of the 77 thyroid cancers resected, 22 were FVPTC and 17 were microcarcinomas (1 medullary carcinoma, 16 PTC). The rates of "unsatisfactory," "cancer," "suspicious for cancer," "follicular neoplasm," and "benign" cytology were 0.7%, 4.4%, 2.6%, 10.2%, and 82.1%, respectively and the cancer yields at surgery were 98%, 81.8%, 15.8%, and 0% respectively. Of the 1127 satisfactory FNAs in the series with a 2- to -6 years of clinical follow-up, a false-negative rate of 0% and a false-positive rate of 1.5% were obtained. Of the 169 surgical follow-ups with satisfactory FNAs, a sensitivity of 100%, specificity of 66.7%, positive predictive value of 87.4%, negative predictive value of 100%, and global accuracy of 89.9% were achieved. The paradoxical combination of low unsatisfactory rate and low false-negative rate is attributed to (1) the use of needle puncture without syringe to obtain enough microfollicles from the exceedingly bloody aspirates from follicular neoplasms for a diagnosis, (2) eliciting history of neck trauma to confirm hematomas, (3) using UFP to highlight the grape-like watery clear nuclei of FVPTC evident with a 4x objective, and (4) the precise guidance by ultrasound in sampling microcarcinomas.

Endometrial brush biopsy for the diagnosis of endometrial cancer.

OBJECTIVE: To evaluate a new technique for processing endometrial cytology for the diagnosis and exclusion of endometrial cancer. STUDY DESIGN: All women at risk for endometrial cancer with clinical indications for endometrial biopsy were evaluated by endometrial brush biopsy (Tao Brush, Cook OB-GYN, Bloomington, Indiana) and Pipelle (Cooper Surgical, Shelton, Connecticut) endometrial biopsies during one office visit. Patients were followed longitudinally for the development of endometrial cancer or until undergoing dilatation and curettage or hysterectomy. All comparisons were analyzed using the chi 2 or t test. RESULTS: One hundred one women (mean age, 58; range, 35-86) had endometrial biopsies performed. Median follow-up was > 21 months (range, 3-29). Twenty-two had cancer or atypia, while the remaining had benign diagnoses. When correlated with the final diagnosis, the Tao Brush had 95.5% sensitivity and the Pipelle, 86% sensitivity. Both devices had 100% specificity, positive predictive value of 100% and negative predictive value of 98%. When the results of the two biopsy devices are considered together, the positive and negative predictive value for detecting or excluding endometrial cancer was 100%. Based on 1998 Medicare reimbursements, a simultaneous second office biopsy using the Tao brush could save approximately $67 per case as compared to a sonohistogram and much more when compared to dilatation and curettage. CONCLUSION: Endometrial cancer can be reliably detected and excluded using these two distinct office biopsy devices simultaneously during one office visit. In patients with an indication for endometrial biopsy, no further diagnostic test may be necessary to exclude or diagnose endometrial cancer or atypia.

Application of Ultrafast Papanicolaou stain to body fluid cytology.

OBJECTIVE: To investigate the applicability of the Ultrafast Papanicolaou stain to the cytology of fluids and to compare it with other methods. STUDY DESIGN: Over a 30-month period, 528 unfixed fluids (462 serous effusions, 48 pelvic washings, 16 cyst fluids and 2 bile duct drain fluids) were mixed thoroughly and centrifuged. Two Swedish-style air-dried smears were made and stained with Diff-Quik (Mercedes Medical, Inc., Sarasota, Florida, U.S.A.) and Ultrafast Papanicolaou stain (Richard Allan Scientific, Kalamazoo, Michigan, U.S.A.), and the remaining sediment was fixed in CytoRich Red (TriPath Imaging, Inc., Burlington, North Carolina, U.S.A.), centrifuged onto a 17.5-mm circle with a Hettich cytocentrifuge and stained by the Papanicolaou method. RESULTS: For the 115 malignant fluids, Ultrafast Papanicolaou stain was the preferred method in the 94 non-hematopoietic malignant fluids, Diff-Quik was the preferred method in the 9 hematopoietic malignancies, and CytoRich Red was the preferred preparation in 8 bloody effusions containing rare cancer cells and 4 malignant pelvic washings. The diagnostic turnaround time of smears stained by Ultrafast Papanicolaou stain was < 15 minutes, fast enough for intraoperative consultations. CONCLUSION: It seems that Ultrafast Papanicolaou stain improves the resolution of cytoplasmic and nuclear details of nonhematopoietic cells in body fluids. However, to detect cancer in all types of fluids, Diff-Quik and CytoRich preparations are also required. We now examine three slides per fluid sample, one slide by each of the three techniques.

PNET-like features of synovial sarcoma of the lung: a pitfall in the cytologic diagnosis of soft-tissue tumors.

Fine-needle aspiration (FNA) cytology of soft-tissue tumors is evolving. As more experience is gained, we are becoming aware of potential pitfalls. We describe 2 cases of synovial sarcoma of the lung, primary and metastatic, in patients who had FNA biopsy performed on a lung mass. The cytologic smears showed extremely cellular groups of malignant small round cells, intersected by small blood vessels, with numerous loose single cells, in a background of macrophages and mature lymphocytes. The tumors displayed monomorphic cells forming rosettes and displaying occasional mitoses. A diagnosis of neuroendocrine tumor/primitive neuroepithelial tumor (PNET) was suspected. Furthermore, this suspicion was supported by immunohistochemical stains, which showed positivity for a neuroendocrine marker, Leu 7 (case 1), and for a neural marker, CD 99 (O 13 or HBA 71) (both cases); and negativity for cytokeratins (case 1). The resection specimen of case 1 had mostly tightly packed small round cells, with occasional rosettes, similar to the FNA biopsy, and focal areas composed of spindle cells, organized in a focal fibrosarcoma-like and hemangiopericytoma-like pattern. A balanced translocation between chromosomes X and 18, demonstrated by both karyotyping and fluorescent in situ hybridization (FISH), enabled us to make a diagnosis of synovial sarcoma, which was histologically classified as poorly differentiated. Case 2 was a metastatic biphasic synovial sarcoma of the arm, with a prominent epithelial component. Synovial sarcoma, when composed mainly of small round cells on cytologic smears, is a great mimicker of neuroendocrine/PNET tumors, with light microscopic and immunohistochemical overlap. Awareness of this potential pitfall may aid in preventing a misdiagnosis. Its recognition is of major concern, especially for the poorly differentiated variant, because it is associated with a worse prognosis.

Fine-needle aspiration cytology of Schiller-Duval bodies of yolk-sac tumor.

The objective of this study was to find the cytological equivalent of Schiller-Duval bodies seen in histology in order to employ the findings as a diagnostic clue for yolk-sac tumor (YST) in aspiration biopsy. A YST, aspirated from the retroperitoneal lymph node of a 29-yr-old Asian man with testicular complex mature teratoma, contained numerous Schiller-Duval bodies on cell block sections. In the corresponding smears, a network of tumor cell-coated, anastomosing, thin-walled, rigid hematopoietic channels with dramatic changes in diameter was observed, in agreement with the diagram based on the reconstruction of 48 serial plastic sections done by Kazancigil et al. (Am J Cancer 1940;40:199-212). In addition, some of the hematopoietic channels folded over several times like a long, narrow balloon pinched at its joints, i.e., "balloon animal"-like. Many segments of the allantoic viteline vascular network, fractured by smearing, exhibited three- or four-way branchings, bulbous protrusions, and cauliflower-like branchings of widely variable diameter.