【 Abstract 】 Objective To study the mRNA expression of BC047440 gene in multiplicate malignant tumor tissues and the corresponding adjacent tissues, and to investigate its roles in the carcinogenesis and development of malignant tumors. Methods Forty-eight cases of malignant tumor tissues and their adjacent non-cancerous tissues were examined. The mRNA expression of BC047440 gene in those tissues of liver cancer, cholangiocarcinoma, gastric cancer, carcinoma of large intestine, glioma, and breast cancer were measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results ① The mRNA expressions of BC047440 gene in liver cancer, gastric cancer, cholangiocarcinoma and carcinoma of large intestine were significantly higher than those in their adjacent non-cancerous tissues (Plt;0.05 or 0.01). BC047440 gene were highly expressed in both glioma and its adjacent tissues (Pgt;0.05), and poorly expressed in both breast cancer and its adjacent tissues (Pgt;0.05). ② There were close relationships between BC047440 gene expression and clinicopathologic findings of liver cancer, including tumor size and portal vein invasion (Plt;0.05). ③ There were also close relationships between BC047440 gene expression and different clinical stages in alimentary canal cancers (Plt;0.05). Conclusion The over expression of BC047440 gene may be related with the growth, infiltration and metastasis of some malignant tumors, including liver cancer, cholangiocarcinoma, gastric cancer, carcinoma of large intestine and glioma.
Histological studies and morphometry quantitative analysis have been performed on trial rabbit’s dilated common bile duct(CBD),which does not dilate simultaneously.The results shows:①Epithelia of rabbit’s CBD have a ber reparable function,which is fairly significant to the prevention of bile duct’s further injure under the pathogenic situation.②The smooth muscle cell(SMC)of the CBD is the histological basis of contraction,some SMC can be seen in contracting state under light microscope.This indicates that the SMC in rabbit’s CBD possess contracting function.③The collagenous and elastic fibers have the normal histological morphometric characteristics and quantity in it’s dilatation process,and no breekdown and degeneration of the fibers can be detected.Because of the morphological structure of these sections is quite similiar with normal ones,theoretically,we suspect that when pathological change of bile duct’s distal portion is relieved and the bile pressure is normal again.It is possible for this dilating bile duct to return to its formal shape and size.
The new mechanisms of automaticity controlled by the calcium and membrane clocks in sinoatrial node are helpful to revealing the sinus arrhythmia, but the present calcium dynamic model is only on the single cell level. In the present study, a central and peripheral single cell model was developed, and by exponentially changing the cell membrane capacitance, size, conductance and gap junction from the center to the periphery, a two-dimensional inhomogeneous sinus and atrial model was created on the basis of the anatomical structure. Five-point difference and finite element methods were used to process the internal grids and the borders. Irregular borders were defined by creating segment trial functions. Quantitative experiments suggested the consistency of the central and peripheral action potentials with related reports in amplitude, cycle length, maximum diastolic potential and upstroke velocity. Functions of the calcium and membrane clocks on the leading pacemaker site and upstroke velocity as well as the effects of the atrial premature beat on the sinus automaticity were also in good agreement with those in other studies. The developed model is helpful for deeply studying relative roles of the calcium and membrane clocks in automaticity and the relations with electrical activities in atrium. At the same time it will lay the foundation for building three-dimensional sinus and atrial organic models.
Objective To analyze the variation of intestinal microflora in patients with colorectal cancer by SYBR GreenⅠreal-time fluorescence quantitative PCR and reveal the role and significance of intestinal microflora in the colorectal cancer-associated molecular pathogenesis. Methods A set of 16S rRNA gene group of species-specific primers for Bifidobacterium spp., Lactobacillus group, Escherichia coli, and ddl gene-targeted species-specific primers for Enterococcus faecalis and feces Enterococcus were designed. Patients with colorectal cancer (colorectal cancer group, n=30) and healthy volunteers (normal control group, n=30) were included and whose feces were collected to extract bacterial genome DNA. SYBR GreenⅠ real-time fluorescence quantitative PCR was used to analyze the five mentioned bacterial amounts. Results Level of Bifidobacterium spp. (4.52±0.49) and Lactobacillus group (5.46±0.12) in colorectal cancer group were significantly lower than those （9.25±0.83 and 7.45±0.37） of normal control group (Plt;0.05), whereas levels of Escherichia coli (5.82±0.47), Enterococcus faecalis (10.6±0.30) and feces Enterococcus (5.74±0.16) in colorectal cancer group were significantly higher than those （4.68±0.32, 4.95±0.24, and 5.03±0.43） of normal control group (Plt;0.05). Conclusions The fecal microflora composition of patients with colorectal cancer is significantly decreased in Bifidobacterium spp. and Lactobacillus group, whereas increased in Escherichia coli, Enterococcus faecalis, and feces Enterococcus. These data underline that the occurrence and progress of colorectal cancer may be related to intestinal microflora.
ObjectiveTo establish a method that can eliminate the pollution of endogenous nucleic acid in the real-time quantitative polymerase chain reaction (PCR) reaction system, which can be used to reduce or eliminate the false positive rate of real-time PCR assay in detection of postoperative intracranial bacteria infection.MethodsAt first, eliminated the pollution of endogenous nucleic acid in the real-time PCR reaction system. Then, with mixed bacteria DNA as a template, multiple PCR was used to specifically identify the gram-negative bacteria. Meanwhile, evaluated the text line and sensitivity of the multiple PCR after eliminating pollution in detecting the DNA of the mixed bacteria.ResultsThe method established could quickly eliminate the pollution of endogenous nucleic acid in the real-time PCR reaction system, and it didn’t affect the Taq enzyme activity and the amplification efficiency in PCR system, with the minimum detection limit of 102 CFU/mL (Staphylococcus aureus and Pseudomonas aeruginosa), which was the same to the culture method. The enzyme cutting method had no significant effect on the activity and amplification efficiency of the enzyme in PCR system, It had no effect on PCR reaction system and primer specificity (Ct=32, ΔRn=200). However, the filtration method significantly reduced the PCR amplification efficiency (Ct=32, ΔRn=150).ConclusionsThis method can easily and rapidly eliminate the pollution of endogenous nucleic acid in the real-time PCR reaction system, and greatly reduce the false positive of PCR detection. It is able to timely and accurately diagnose the intracranial bacteria infection, which is significant for clinical testing.
More and more medical devices can capture different features of human body and form three dimensional (3D) images. In clinical applications, usually it is required to fuse multiple source images containing different and crucial information into one for the purpose of assisting medical treatment. However, traditional image fusion methods are normally designed for two dimensional (2D) images and will lead to loss of the third dimensional information if directly applied to 3D data. Therefore, a novel 3D magnetic image fusion method was proposed based on the combination of newly invented beyond wavelet transform, called 3D band limited shearlet transformand (BLST), and four groups of traditional fusion rules. The proposed method was then compared with the 2D and 3D wavelet and dual-tree complex wavelet transform fusion methods through 4 groups of human brain T2* and quantitative susceptibility mapping (QSM) images. The experiments indicated that the performance of the method based on 3D transform was generally superior to the existing methods based on 2D transform. Taking advantage of direction representation, shearlet transform could effectively improve the performance of conventional fusion method based on 3D transform. It is well concluded, therefore, that the proposed method is the best among the methods based on 2D and 3D transforms.
Trabecular microstructure is an important factor in determining bone strength and physiological function. Normal X-ray and computed tomography (CT) cannot accurately reflect the microstructure of trabecular bone. High-resolution peripheral quantitative computed tomography (HR-pQCT) is a new imaging technique in recent years. It can qualitatively and quantitatively measure the three-dimensional microstructure and volume bone mineral density of trabecular bone in vivo. It has high precision and relative low dose of radiation. This new imaging tool is helpful for us to understand the trabecular microstructure more deeply. The finite element analysis of HR-pQCT data can be used to predict the bone strength accurately. We can assess the risk of osteoporosis and fracture with three-dimensional reconstructed images and trabecular microstructure parameters. In this review, we summarize the technical flow, data parameters and clinical application of HR-pQCT in order to provide some reference for the popularization and extensive application of HR-pQCT.
Quantitative susceptibility mapping (QSM) can provide tissue susceptibility information and has been adapted for clinical research and diagnosis. QSM of monkey brain in vivo at 9.4 T has not been demonstrated so far. In this study 9.4 T in vivo monkey brain QSM was performed with 200 μm isotropic high-resolution. It was found that the inherent singularity problem for QSM diverged significantly at ultra-high image resolution during regularization process and resulted in severe image artifacts. The K-space division (TKD) was applied to eliminate the artifacts, with an optimal threshold level between 0.2 and 0.3. High resolution QSM of monkey brain in vivo can thus provide a novel tool for brain research.
Objective To observe whether Cyclo-RGDfK (Arg-Gly-Asp-D-Phe-Lys) could enhance the adhesion of myofibroblast to decellularized scaffolds and upregulate the expression of Integrin αVβ3 gene. Methods Myofibroblast from the rat thoracic aorta was acquired by primary cell culture. The expression of Vimentin and α-smooth muscle actin(α-SMA) has been detected by immunoflurescent labeling. Decellularized valves have been randomly divided into three groups (each n=7). Group A (blank control): valves do not receive any pretreatment; Group B: valves reacted with linking agent NEthylN(3dimethylaminopropyl)carbodiimide hydrochloride (EDC) for 36 hours before being seeded; Experimental group: Cyclo-RGD peptide has been covalently immobilized onto the surface of scaffolds by linking agent EDC. The fifth generation of myofibroblast has been planted on the scaffolds of each group. The adhesion of myofibroblast to the scaffolds was evaluated by HE staining and electron scanning microscope. The expression of Integrin αVβ3 was quantified by halfquantitative reverse transcriptionpolymerase china reaction (RT-PCR). Results We can see that myofibroblast has exhibited b positive staining for Vimentin and α-SMA. Besides, it has been shown that the expression of Integrin αVβ3 was much higher in the experimental group than that of the group A and group B(Plt;0.05). There was no statistically difference in group A and group B (P=0.900). Conclusion RGD pretreatment does enhance the adhesive efficiency of seeding cells to the scaffolds and this effect may be related to the upregulation of Integrin αVβ3.
Objective To detect traces of microRNAs (miRNAs) in plasma and assess the expression stability of two common reference genes by stem-loop and poly A polymerase (PAP) real-time quantitative polymerase chain reaction (PCR) method, as miRNAs are the new bio-markers of tumor diagnosis and molecular targeted therapy, and its quantitative research is very important. Methods We extracted miRNAs from plasma of adult Sprague Dawley (SD) rats’ plasma, and detected the expressions of rno-miR-200b-3p and rno-miR-126-3p with stem-loop and PAP real-time PCR quantitative method, with rno-miR-103a-3p and U6 as internal controls. All the results were evaluated by 2△Ct method. Results Compared with PAP method, the stem-loop method reduced Ct value by 2-4 cycles and improved sensitivity by 10 times. In PAP method, the melting curve showed two peaks, a main peak and a small non-specific peak. Yet the melting curve of stem-loop method demonstrated a single specific peak. Furthermore, we validated the stability of internal references in the two real time PCR methods. U6 presented a more stable Ct value than rno-miR-103 in adult SD rats’ plasma samples. Conclusions Stem-loop real-time PCR is recommended as a major way to detect some samples with a low concentration of miRNAs, owing to its high accuracy and sensitivity. However, if a large number of tissue samples is going to be detected, PAP real-time PCR is more suitable and convenient than stem-loop method. U6 is more stable and repeatable than rno-miR-103a-3p as the reference gene to evaluate the semi-quantitative consequence of miRNAs.
