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STATEMENT OF PROBLEM Digital light processing (DLP), continuous liquid interface printing (CLIP), and stereolithography (SLA) technologies enable 3-dimensional (3D) printing of surgical guides. However, how their accuracy compares and how accuracy may affect subsequent steps in guided surgery is unclear. PURPOSE The purpose of this in vitro study was to investigate the fabrication and seating accuracy of surgical guides printed by using DLP, SLA, and CLIP technologies and evaluate the positional deviation of the osteotomy site and placed implant compared with the digital implant plan. MATERIAL AND METHODS Twenty-one polyurethane models were divided into 3 groups and used to plan implants and design surgical guides. The guides were fabricated by using DLP, SLA, or CLIP 3D printers (n=7) and scanned, and the scan file was compared with the digital design file to analyze the fabrication accuracy at the intaglio and overall external surfaces using root mean square (RMS) values. The triple scan protocol was used to evaluate the seating accuracy of the guides on their respective models. Osteotomies were prepared on models by using the guides followed by a microcomputed tomography image of each osteotomy. The implants were placed through the guides, the scan bodies were tightened to implants, and the models were scanned to obtain the images of placed implant position. Osteotomy and placed implant images were used to calculate the entry point, apex, and long axis deviations from the planned implant position with a software program. A 2-way repeated-measures ANOVA of the RMS data was used to analyze printing and seating trueness, and homogeneity of variance analyses were used at each surface for precision. A 3-way repeated-measures ANOVA was used to analyze distance deviations over the stages (osteotomy and final implant) and locations studied, and a 2-way repeated-measures ANOVA was used for angular deviations. Homogeneity of variance analyses were performed for precision (α=.05). RESULTS The 3D printer type significantly affected the trueness of the guide at the intaglio surface (P

Alteration of DNA methylation is an important event in pathogenesis and progression of hepatocellular carcinoma (HCC). DNA methyltransferase (DNMT) 1, the foremost contributor in DNA methylation machinery, was revealed elevated in HCC and significantly correlates with poor prognosis. However, the transcriptional regulation of DNMT1 in HCC remains unknown.Real-time PCR and immunohistochemistry were performed to detect DNMT1 and zinc finger transcription factor 191 (ZNF191) expressions in HCCs. Transcription activity of DNMT1promoter was analyzed with Luciferase reporter activity assay. The binding capacity of ZNF191 protein to DNMT1 promoter was examined with chromatin immunoprecipitation-qPCR (ChIP-qPCR) and electrophoretic mobility shift assay (EMSA). DNA methylation level of hepatoma cells was detected with Methylation array.ZNF191 can regulate DNMT1 mRNA and protein expression positively, and increase the transcription activity of the DNMT1 promoter. ChIP-qPCR and EMSA revealed that ZNF191 protein directly binds to the DNMT1 promoter at nt-240 AT(TCAT)ZNF191 is a novel transcription regulator for DNMT1, and the pro-proliferation effect of ZNF191/DNMT1/p-AKT axis in hepatoma cells implies that ZNF191 status in HCCs may affect the therapeutic effect of DNMTs inhibitors and PI3K inhibitors for precise treatment of the disease.

Fula Sub-Basin, located in NE of Muglad Basin, is the most important production base for block 6, which is one of the important overseas project of CNPC. After more than 20 years’ exploration and development, big and shallower traps such as Bentiu/Aradeiba reservoir have been discovered and drilled for most area, remained traps are limited with small area, deep targets, and complex structure which are difficult to identify. How to exploring deeper targets as Abu Gabra has become more and more urgent. Many challenges are faced for detailed exploration of Abu Gabra such as poor data quality in deep parts and complex fault system identification etc. Some specific techniques and methods have been tried and tested to identify new traps, including target filter processing for deep parts, well-controlled fault point identification integrated with detailed well correlation, fault polygon generation with multiple attributes etc., and finally a combined work flow for detailed exploration of complex faulted trap has been generated. These techniques have been applied in Fula and other Sub-basin in block 6 and a series of new faulted traps have been discovered. Drilling candidates are selected from them through detailed trap appraisal and good results come out, with 11 successful wells from 12 candidates, which indicate that integrated fault identification techniques are effective for detailed exploration and development of rifting sub-basins in Muglad basin and similar basin.

N gas field is one of the main gas suppliers with an oil-rim at the slope in the south Sumatra basin. Complicated by high CO2 in the faulted condensated reservoir, the distribution of gas, oil and water, and residual fluid were not easy to explore, which is essential for drilling future wells. This paper builds a four-step workflow to determine the fluid distribution to obtain maximum production. The first step in our workflow was to finely delineate the faults by the techniques of coherence and dip detection, which built basis for oil–gas compartment division. Second, petrophysical modeling was carried in geology software platform to compute the properties. The reservoir and fluid cutoffs can be cross-plotted between measured properties for the non-core wells. Then the fluid systems were delineated in vertical and areal directions according to DST and fluid cutoffs. The vertical and areal CO2 samples were finely compared to determine the compartment boundaries in consideration of fault combinations. Finally, the residual oil and gas potential were simulated after history match in well and field level. Five proposed wells have been drilled and gotten the 5% increase compared with the planned rate production. The result showed this workflow is useful for improving production and field performances.

Fula sub-basin is located in the northeastern part of the Muglad Basin in Sudan. Seismic attribute analysis and seismic inversion are employed to perform reservoir prediction in Fula3D area. On the basis of reservoir prediction results, the stratigraphic traps are identified in this area. According to sequence stratigraphic study of Fula sub-basin, AG2 in SW of Fula3D area was selected to carry out reservoir prediction and identification of stratigraphic traps. AG2 can be divided into five sub-layers(AG2-A – AG2-E). Based on the detailed interpretation of AG2-D and AG2-E, seismic attribute analysis and seismic inversion were carried out in Fula3D area. Among seismic attributes analyze, the results of arc length indicated the interbedded sandstones and shale in AG2-D and AG2-E. By applying CSSI (Constrained Sparse Spike Inversion), the P-impedance volume is obtained from seismic data of Fula3D area. According to the seismic inversion section across the well F-1, F-4 and F-5, the thickness changes and sandstone pinch-out exist along the structural belts. Stratal slices of seismic inversion results show that sandstones mainly developed in the lower part of AG2 in the southern part of Fula3D area. The structural analysis show that the SW of Fula3D area is a complex faulted terrace controlled by four faults. F-1 faulted block is considered to be a favorable target through comprehensive analysis of structural, seismic attribute, inversion volume, sedimentary facies and hydrocarbon accumulation conditions. Therefore, we proposed the stratigraphic pinch-out area in the F-1 faulted block is a favorable area for stratigraphic trap.

Treacher Collins syndrome is an autosomal dominant craniofacial malformation mainly caused by mutations in the TCOF1 gene. Few cases have been observed in the Chinese population. Herein, the authors report the mutational analysis of TCOF1, GSC, and HOXA2 to determine the mutational features of the 3 genes in Chinese patients with Treacher Collins syndrome. Genomic DNA of the patients and their parents was extracted from peripheral blood following a standard protocol. DNA sequencing analysis was performed on all exons and the exon-intron borders of TCOF1, GSC, and HOXA2 in addition to the 1200-bp upstream of TCOF1. Four novel single nucleotide polymorphisms were detected in TCOF1, one of which was in the promoter region. Mutations in GSC and HOXA2 were not found in the 3 patients. Our results suggest the possibility of genetic heterogeneity or different mechanisms leading to the disease. Further functional study of the alteration is necessary to obtain more definitive information.

Total internal reflection (TIR) is useful for interrogating physical and chemical processes that occur at the interface between two transparent media. Yet prism-coupled TIR imaging microscopes suffer from limited sensing areas due to the fact that the interface (the object plane) is not perpendicular to the optical axis of the microscope. In this paper, we show that an electrically tunable lens can be used to rapidly and reproducibly correct the focal length of an oblique-incidence scanning microscope (OI-RD) in a prism-coupled TIR geometry. We demonstrate the performance of such a correction by acquiring an image of a protein microarray over a scan area of 4 cm² with an effective resolution of less than 20 microns. The electronic focal length tuning eliminates the mechanical movement of the illumination lens in the scanning microscope and in turn the noise and background drift associated with the motion.

Human tissue factor pathway inhibitor-2 (Tfpi-2) is an extracellular matrix-associated Kunitz-type serine proteinase inhibitor and plays an important role in various cellular processes. We have previously shown that zebrafish Tfpi-2 (zTfpi-2) mainly expressed in the brain and heart of zebrafish, and it is involved in the development of central nervous system. Here, we identified zTfpi-2 as an evolutionarily conserved protein essential for zebrafish heart development, as embryos depleted of zTfpi-2 failed to undergo cardiogenesis. Changes of cardiogenic markers, vmhc, amhc and bmp4, confirmed zTfpi-2 knockdown caused cardiac defects, including retrenched ventricle, enlarged atrium and malformation of atrioventricular boundary. The sarcomeric organization was also disrupted by embryonic depletion of zTfpi-2, thus establishing the functional role of zTfpi-2 in cardiac contractility. In addition, hematopoietic defects were detected in the zTfpi-2-deficiency embryos. Importantly, injection of ztfpi-2 mRNA attenuated those cardiac and hematopoietic defects. Taken together, this study demonstrated a critical role of zTfpi-2 during embryonic cardiac development, as well as an important regulator of hematopoiesis.

Microtia is a complicated congenital anomaly with a genetic and environmental predisposition, and the molecular events underlying this disease are not fully understood. MicroRNAs (miRNAs) are a class of 20-22 nucleotide non-coding RNAs that function to control post-transcriptional gene expression. We want to find the miRNA expression profiling of microtia by using Affymetrix GeneChip(®) miRNA 2.0 Arrays.We selected 9 microtia cartilages and 3 normal controls for GeneChip(®) miRNA 2.0 Arrays analysis. The altered miRNA were analyzed by poly (A) RT-PCR from 58 microtia samples and 16 normal controls. We predicted the target genes of miRNAs by bioinformatics and RT-PCT was used to confirm the target genes.We found 11 miRNAs with significantly altered expression in the microtic group compared to the normal controls, which included 6 up-regulated miRNAs and 5 down-regulated miRNAs. These miRNAs were further examined using poly (A) RT-PCR analysis, we found that miR-451 and miR-486-5p were significantly up-regulated and miR-200c was significantly down-regulated in the microtic group compared to the normal controls (p0.05). Several complementary target messenger RNAs (mRNAs) had been predicted. OSR1, the target gene of miR-451 and miR-200c, was significantly up-regulated (p0.01); TRPS1, the target gene of miR-200c, was significantly down-regulated in the microtic group compared to the controls (p0.0001).The reduction in miR-200c expression and the accretion of miR-451 and miR-486-5p expression in microtic samples could be possible causes of the abnormal development of the external ear. OSR1 and TRPS1, as the complementary target mRNAs, may play important roles during the development of the external ear. Further studies are still needed to identify the miRNA target genes and to determine their function in the pathogenesis of microtia. This is the first report of a relationship between miRNAs and microtia.

Objective Microtia is defined as a developmental malformation characterized by a small, abnormal shaped auricle, with atresia or stenosis of the auditory canal. Genes responsible for nonsyndromic microtia have remained elusive. We therefore report a mutational analysis of GSC, HOXA2 and PRKRA in 106 congenital microtia patients without any combined malformation to explore the relationship between GSC, HOXA2, PRKRA and nonsyndromic microtia. Methods A total of 106 patients with a clinical diagnosis of congenital microtia and a control group (100 unaffected controls) were recruited through the Eye and ENT Hospital of Fudan University in China. Genomic DNA was extracted following a standard protocol. DNA sequencing analysis was performed in all exons and the exon-intron borders of GSC, HOXA2 and PRKRA. Results We identified 5 genomic variants in GSC, HOXA2 and PRKRA. As to the GSC, we obtained a reported variant g.994C > T in exon 2, which resulted in no change of protein. Our results revealed that g.994C > T was also detected in 10 control cases. We also detected 2 novel variants, g.90G > A and g.114A > C, in the 5′UTR of HOXA2. No class 5 or 4 genomic variant of PRKRA was identified in our microtia patients. Additionally, two previously reported SNVs in GSC and PRKRA were also presented. Conclusions We suggest that g.994C > T is a new SNV, which is different from the previous report. Further study is needed to prove the function of 2 novel variants in the 5′UTR of HOXA2, and to explore the possible mechanism of these variants in the occurrence of microtia.

In reverse-phase protein arrays (RPPA), one immobilizes complex samples (e.g., cellular lysate, tissue lysate or serum etc.) on solid supports and performs parallel reactions of antibodies with immobilized protein targets from the complex samples. In this work, we describe a label-free detection of RPPA that enables quantification of RPPA data and thus facilitates comparison of studies performed on different samples and on different solid supports. We applied this detection platform to characterization of phosphoserine aminotransferase (PSAT) expression levels in Acanthamoeba lysates treated with artemether and the results were confirmed by Western blot studies.

Small-molecule microarray (SMM) is an effective platform for identifying lead compounds from large collections of small molecules in drug discovery, and efficient immobilization of molecular compounds is a pre-requisite for the success of such a platform. On an isocyanate functionalized surface, we studied the dependence of immobilization efficiency on chemical residues on molecular compounds, terminal residues on isocyanate functionalized surface, lengths of spacer molecules, and post-printing treatment conditions, and we identified a set of optimized conditions that enable us to immobilize small molecules with significantly improved efficiencies, particularly for those molecules with carboxylic acid residues that are known to have low isocyanate reactivity. We fabricated microarrays of 3375 bioactive compounds on isocyanate functionalized glass slides under these optimized conditions and confirmed that immobilization percentage is over 73%.

Background MicroRNAs (miRNAs) are noncoding RNAs involved in posttranscriptional regulation of gene expression in cancer and provide new perspectives on the development of laryngeal squamous cell carcinoma (SCC). Methods miRNA expression of 6 pairs of laryngeal SCC and adjacent normal tissues was screened using miRNA array. Laser capture microdissection was applied to isolate a homogeneous group of cells from laryngeal SCC samples. The results of miRNA array analysis were validated in 48 pairs of laryngeal SCC and adjacent normal tissues using quantitative RT-PCR. Results Twenty-nine differentially expressed miRNAs were detected in the 6 pairs of laryngeal SCC, of which 6 were confirmed, including upregulation of miR-21, miR-93, miR-205, and miR-708 and downregulation of miR-125b and miR-145. Their putative target genes were predicted using 3 online software programs. Conclusion These differentially expressed miRNAs may play a role in tumorigenesis and progression in laryngeal SCC and offer new angles for further investigations into the function of miRNAs. © 2012 Wiley Periodicals, Inc. Head Neck, 2013

Velocardiofacial syndrome (VCFS) is a disease in human with an expansive phenotypic spectrum and diverse genetic mechanisms mainly associated with copy number variations (CNVs) on 22q11.2 or other chromosomes. However, the correlations between CNVs and phenotypes remain ambiguous. This study aims to analyze the types and sizes of CNVs in VCFS patients, to define whether correlations exist between CNVs and clinical manifestations in Chinese VCFS patients. In total, 55 clinically suspected Chinese VCFS patients and 100 normal controls were detected by multiplex ligation-dependent probe amplification (MLPA). The data from MLPA and all the detailed clinical features of the objects were documented and analyzed. A total of 44 patients (80.0%) were diagnosed with CNVs on 22q11.2. Among them, 43 (78.2%) presented with 22q11.2 heterozygous deletions, of whom 40 (93.0%) had typical 3-Mb deletion, and 3 (7.0%) exhibited proximal 1.5-Mb deletion; no patient was found with atypical deletion on 22q11.2. One patient (1.8%) presented with a 3-Mb duplication mapping to the typical 3-Mb region on 22q11.2, while none of the chromosomal abnormalities in the MLPA kit were found in the other 11 patients and 100 normal controls. All the 43 patients with 22q11.2 deletions displayed characteristic face and palatal anomalies; 37 of them (86.0%) had cognitive or behavioral disorders, and 23 (53.5%) suffered from immune deficiencies; 10 patients (23.3%) manifested congenital heart diseases. Interestingly, all patients with the characteristic face had 22q11.2 heterozygous deletions, but no difference in phenotypic spectrum was observed between 3-Mb and 1.5-Mb deletions. Our data suggest that the characteristic face can be used as a key indicator for direct diagnosis of 22q11.2 deletions in Chinese VCFS patients.

Oblique-incidence reflectivity difference (OI-RD) is a form of polarization-modulation ellipsometry that measures properties of thin films on a solid surface through the change in polarization state of light upon reflection from the surface. The measurement accuracy depends on the precision of the phase modulation amplitude and azimuthal alignments of key polarizing optical elements and, thus, requires careful calibration. In the present work, we describe robust methods of such calibrations that enable precise determination of the modulation amplitude and static retardation of a phase modulator and azimuths of key polarizing optics in an OI-RD system.

Background miRNAs are a group of small RNA molecules regulating target genes by inducing mRNA degradation or translational repression. Aberrant expression of miRNAs correlates with various cancers. Although miR-135a has been implicated in several other cancers, its role in breast cancer is unknown. HOXA10 however, is associated with multiple cancer types and was recently shown to induce p53 expression in breast cancer cells and reduce their invasive ability. Because HOXA10 is a confirmed miR-135a target in more than one tissue, we examined miR-135a levels in relation to breast cancer phenotypes to determine if miR-135a plays role in this cancer type. Methods Expression levels of miR-135a in tissues and cells were determined by poly (A)-RT PCR. The effect of miR-135a on proliferation was evaluated by CCK8 assay, cell migration and invasion were evaluated by transwell migration and invasion assays, and target protein expression was determined by western blotting. GFP and luciferase reporter plasmids were constructed to confirm the action of miR-135a on downstream target genes including HOXA10. Results are reported as means ± S.D. and differences were tested for significance using 2-sided Student"s t-test. Results Here we report that miR-135a was highly expressed in metastatic breast tumors. We found that the expression of miR-135a was required for the migration and invasion of breast cancer cells, but not their proliferation. HOXA10, which encodes a transcription factor required for embryonic development and is a metastasis suppressor in breast cancer, was shown to be a direct target of miR-135a in breast cancer cells. Our analysis showed that miR-135a suppressed the expression of HOXA10 both at the mRNA and protein level, and its ability to promote cellular migration and invasion was partially reversed by overexpression of HOXA10. Conclusions In summary, our results indicate that miR-135a is an onco-miRNA that can promote breast cancer cell migration and invasion. HOXA10 is a target gene for miR-135a in breast cancer cells and overexpression of HOXA10 can partially reverse the miR-135a invasive phenotype.

Background The purpose of this study is to evaluate the prognostic value of TFPI-2 expression in breast cancer patients through examining the correlation between TFPI-2 expression and breast cancer clinicopathologic features. Methods Immunohistochemical staining combined with digital image analysis was used to quantify the expression of TFPI-2 protein in breast tumor tissues. For evaluation of the prognostic value of TFPI-2 expression to each clinicopathologic factor, Kaplan-Meier method and COX’s Proportional Hazard Model were employed. Results TFPI-2 expression was significantly correlated with tumor size, lymph node metastasis, histologic grade, clinical stage, and vessel invasion. More importantly, TFPI-2 expression was also associated with disease-free survival (DFS) of breast cancer patients. We found that patients with high TFPI-2 expression had longer DFS compared with those with low or negative expression of TFPI-2 (P P Conclusions Low or negative expression of TFPI-2 is associated with breast cancer progression, recurrence and poor survival outcome after breast cancer surgery. TFPI-2 expression in breast tumors is a potential prognostic tool for breast cancer patients.