Objective To investigate the detection of peritoneal free cancer cells and its clinical significance. Methods The peritoneal free cancer cells, the positive rates of CK20 protein and CK20 mRNA expressions of peritoneal lavage fluid were detected by peritoneal lavage cytology (PLC), flow cytometry (FCM) and real-time fluorescent quantitative RT-PCR in 50 cases of gastric cancer patients, respectively. The sensitivity of three kinds of detection method to peritoneal free cancer cells was compared. Results The positive rates of peritoneal free cancer cells, CK20 protein and mRNA expression of peritoneal lavage fluid were 20.0% (10/50), 36.0% (18/50) and 58.0% (29/50), respectively. The positive rate of CK20 mRNA expression detected by real-time fluorescencequantitative RT-PCR in peritoneal lavage fluid was significantly higher than those of the CK20 protein expression detected by FCM and peritoneal free cancer cells detected by PLC (Plt;0.05 or Plt;0.001). The difference of positive rate of CK20 protein expression and peritoneal free cancer cells was not significant (Pgt;0.05). The positive rate of CK20 mRNA expression of peritoneal lavage fluid was related to the tumor invasion depth, differentiation degree, TNM stage, and lymph node metastasis (Plt;0.05). Conclusion Real-time fluorescence quantitative RT-PCR is an effective method for the detection of peritoneal free cancer cells.
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.
【 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.
As the incidence of osteoporosis (OP) is increasing year by year, high morbidity and mortality caused by osteoporotic fractures have become major problems of health in China and over the world. Quantitative measurement of bone using 99mTc-methylene diphosphonate (99mTc-MDP) provides global or local information of skeletal metabolism or transformation. In this paper, we make a brief review on the quantitative measurement of bone using 99mTc-MDP and expect to provide guidance for clinical diagnosis and treatment.
Immuno-fluorescence technique can qualitatively determine certain nuclear translocation, of which NF-κB/p65 implicates the activation of NF-κB signal pathways. Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-κB/p65 by computing relative fluorescence units in nuclei and cytoplasm. We verified the quantitative analysis by Western Blot. When we applied the software to analysis of nuclear translocation in lipopolysaccharide (LPS) induced (0.5 h, 1 h, 2 h, 4 h) primary human umbilical vein endothelial cells (HUVECs), we found that nuclear translocation peak showed up at 2h as with calculated Western blot verification results, indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence.
Quantitative assessment of the symptoms of Parkinson's disease is the key for precise diagnosis and treatment and essential for long term management over years. The challenges of quantitative assessment on Parkinson's disease are rich information, ultra-low load, long term and large range monitoring in free-moving condition. In this paper, we developed wearable devices with multiple sensors to monitor and quantify the movement symptoms of Parkinson's disease. Five wearable sensors were used to record motion signals from bilateral forearms, legs and waist. A local area network based on low power Wi-Fi technology was built for long distance wireless data transmission. A software was developed for signal recording and analyzing. The size of each sensor was 39 mm×33 mm×16 mm and the weight was 18g. The sensors were rechargeable and able to run 12 hours. The wireless transmission radius is about 45 m. The wearable devices were tested in patients and normal subjects. The devices were reliable and accurate for movement monitoring in hospital.
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 β-secretase is one of prospective targets against Alzheimer's disease (AD). A three-dimensional quantitative structure-activity relationship (3D-QSAR) model of Hydroethylamines (HEAs) as β-secretase inhibitors was established using Topomer CoMFA. The multiple correlation coefficient of fitting, cross validation and external validation were r2=0.928, qloo2=0.605 and rpred2=0.626, respectively. The 3D-QSAR model was used to search R groups from ZINC database as the source of structural fragments. As a result, a series of R groups with relatively high activity contribution was obtained to design a total of 15 new compounds, with higher activity than that of the template molecule. The molecular docking was employed to study the interaction mode between the new compounds as ligands and β-secretase as receptors, displaying that hydrogen bond and hydrophobicity played important roles in the binding affinity between the new compounds and β-secretase. The results showed that Topomer CoMFA and Topomer Search could be effectively used to screen and design new molecules of HEAs as β-secretase inhibitors, and the designed compounds could provide new candidates for drug design targeting AD.
Objective To investigate the gene expression of beta-defensin-4 (mBD-4) and mBD-6 in acute lung injury (ALI) mouse.Methods Sixty adult mice were randomly divided into a control group and a ALI group.ALI was induced by intraperitoneal injection of lipopolysaccharide (LPS) in the ALI group.The control group was treated with same dose of normal saline.The lung tissues were harvested at different time point after stimulation.The expression of mBD-4 and mBD-6 mRNA was measured by real-time quantitative reverse transcription polymerase chain reaction.DNA sequencing was used to confirm the specificity of mBD-4 and mBD-6 cDNA fragment.Results There were no obvious mBD-4 and mBD-6 mRNA expression in mouse lung in the control group at all time points and ALI 6 h group.In the ALI group a marked increasing expression was found on 12 h，1 d and 3 d after LPS stimulation.The mBD-4 mRNA expression was significant higher in the ALI groups of 1 d and 3 d points than that of ALI 12 h group with no obvious difference between each other.There were no significant differences of mBD-6 mRNA expression between ALI groups of 12 h，1 d and 3 d points Conclusion mBD-4 and mBD-6 mRNA is not constitutive expressed in mouse lung and show a up-regulative expression pattern after ALI.
ObjectiveTo investigate the value of proton magnetic resonance spectroscopy (1H-MRS), gradient dual-echo, and triple-echo sequences in the quantitative evaluation of treatment effect of fatty liver at 3.0T MR.MethodsThirty patients with fatty liver diagnosed by CT or ultrasound who admitted in Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital between August 2017 and May 2018, were enrolled and undergone gradient dual-echo, triple-echo, and 1H-MRS examination before and 3 months after treatment. The fat index (FI) and relative lipid content (RLC) were measured. Fatty liver index (FLI) was calculated from blood biochemical indicators, waist circumference, and BMI at the same time. With the reference standard of FLI, the results before and after treatment measured from MRI were analyzed.ResultsThere were significantly differences of FLI, FIdual, FItriple, and RLC before and after treatment (t=5.281, P<0.001; Z=–3.651, P<0.001; Z=–3.630, P<0.001; Z=–4.762, P<0.001), all indexes decreased after treatment. FIdual and FItriple were positively correlated with FLI before (rs=0.413, P=0.023; rs=0.396, P=0.030) and after treatment (rs=0.395, P=0.031; rs=0.519, P=0.003), the highest correlation factor was FItriple to FLI after treatment. There were no significant correlation between RLC and FLI before and after treatment (P>0.05).ConclusionsIt is feasible to quantitatively evaluate the treatment effect of fatty liver by using 1H-MRS, gradient dual-echo, and triple-echo sequences. Gradient triple-echo sequences has better accuracy, which is technically easy to implement and more suitable for clinical development.