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 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.
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.
Objective To scientifically evaluate the nosocomial infection prevention and control ability of respiratory infectious diseases in general hospitals, and to construct a set of quantitative assessment system for the prevention and control ability of respiratory infectious diseases in general hospitals. Methods Papers, standards and guidelines online related to respiratory hospital infections published between January 1, 2010 and December 31, 2023 were selected, and infection control experience was summarized to build a pool of evaluation pionts. Then, this study used experts consultation to select the evaluation pionts, to calculate the weight coefficient and reached a consensus on the quantitative evaluation methods of each evaluation point. Results A total of 27 articles were included. The evaluation system included 17 evaluation points in 4 categories: “Basic management capacity” “Basic conditions of facilities and equipments” “The prevention and control capacity of nosocomial infection” and “The emergency response capacity”. Each evaluation point contained 3 quantitative evaluation indicators. The Cronbach’s α coefficients of the expert questionnaire consultation were 0.914 and 0.883, respectively. The scale-level content validity index was 0.932, and the item-level content validity index ranged from 0.823 to 1. Conclusions The evaluation system constructed in this study can be used for quantitative evaluation and quality self-examination of the prevention and control ability of respiratory infectious diseases in general hospitals. It also contributes to the continuous improvement of the quality of nosocomial infection prevention and control.
Intravascular optical coherence tomography (IVOCT) has emerged as a high-resolution and minimal-invasive imaging technique that provides high-speed visualization of coronary arterial vessel walls and clearly displays the vessel lumen and lesions under the intima. However, morphological gray-scale images cannot provide enough information about the tissue components to accurately characterize the plaque tissues including calcified, fibrous, lipidic and mixed plaques. Quantitative IVOCT (qIVOCT) is necessary to provide the physiological contrast mechanisms and obtain the characteristic parameters of tissues with clinical diagnostic value. In this paper, the progress of qIVOCT is reviewed. The current methods for quantitatively measuring optical, elastic and hemodynamic parameters of vessel wall and plaque tissues using IVOCT gray-scale images and raw backscattered signals are introduced and potential development is forecast.
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.
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.
At present the parkinsonian rigidity assessment depends on subjective judgment of neurologists according to their experience. This study presents a parkinsonian rigidity quantification system based on the electromechanical driving device and mechanical impedance measurement method. The quantification system applies the electromechanical driving device to perform the rigidity clinical assessment tasks (flexion-extension movements) in Parkinson’s disease (PD) patients, which captures their motion and biomechanical information synchronously. Qualified rigidity features were obtained through statistical analysis method such as least-squares parameter estimation. By comparing the judgments from both the parkinsonian rigidity quantification system and neurologists, correlation analysis was performed to find the optimal quantitative feature. Clinical experiments showed that the mechanical impedance has the best correlation (Pearson correlation coefficient r = 0.872, P < 0.001) with the clinical unified Parkinson’s disease rating scale (UPDRS) rigidity score. Results confirmed that this measurement system is capable of quantifying parkinsonian rigidity with advantages of simple operation and effective assessment. In addition, the mechanical impedance can be adopted to help doctors to diagnose and monitor parkinsonian rigidity objectively and accurately.
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.
【 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.