Abstract: Objective To investigate the messenger ribonucleic acid (mRNA) expression level of tissue-type plasminogen activator (t-PA) in endothelial cells derived from adult mesenchymal stem cells (MSCs) after fluid shear stress loading which is within the physiological range. Methods After culturing in vitro, bone marrow MSCs of SD rats were seeded on slides.When it come to 80% confluence,26 slides were exposed to 5dyn/cm2 fluid shear stress for 3h in a flow chamber, and then induced to endothelial cells. Among them,13 slides constituted group Ⅰ, and the rest 13 slides set up group Ⅱ, which would be cultured for 3-4d further and passaged in 1∶3. At the same time, control group was set up, which including the cells never exposed to fluid shear stress before the endothelial differentiation. Fluid shear stress were exerting to cells in a specially made flow chamber. The expression level of t-PA mRNA of all groups were measured by real-time fluorescent quantitation reverse transcriptionpolymerase chain reaction (RTPCR). Results After endothelial differentiation for 7 days, the SD rats bone marrow MSCs acquired typical endothelial cell appearance. The t-PA mRNA expression level of group Ⅰ and group Ⅱ have an obviously enhance compared with control group(P＜0.05). The t-PA mRNA expression level of group Ⅱ step down a little (P＞0.05), but it is still significantly higher than that of control group (P＜0.05). Conclusion Fluid shear stress could provide a protective action on the endothelial cells induced from MSCs in vitro, and the effect maintains with the cells passages. This formulates a theoretical foundation to the therapeutics of atherosclerosis and selection of seed cells in vascular tissue engineering.
Objective The usefulness of measurement of nuclear DNA content elevation for diagnosis of early hepatocellular carcinoma was evaluated by a study of 186 patients with liver cirrhosis. Methods Nuclear DNA content was measured using an automatic image analysis system.Results ①Hepatocellular carcinoma was found in 37 patients during 10 years follow-up, the cumulative incidence of hepatocellular carcinoma was 19.89%. ②The incidence of hepatocellular carcinoma increased with the increase of the patterns of α-fetoprotein (AFP), 5c exceeding rate (5cER), FORM PE, but positive predictive value of 5cER was the highest of three parameters, the difference among all groups was significant by the χ2 test (Ｐ＜0.05). ③When 5cER joined AFP for monitoring development of hepatocellular carcinoma, the incidence of hepatocellular carcinoma was 72.00%, which was significantly higher than that of 5cER or AFP alone, the difference between groups was highly significant (Ｐ＜0.01). Conclusion Patients who had 5cER levels of 3%-5% or more, who had transient increases in 5cER or who had both, should be treated as being in a super-highrisk group for hepatocellular carcinoma. Frequent and careful examination by ultrasonography of such patients is recommended. It is important that measurement of 5cER join with AFP in cirrhotic patients monitored for early development of hepatocellular carcinoma.
It is the main method for amplifying the specific gene to use the nucleic acid amplification system to accomplish polymerase chain reaction (PCR). The temperature retard between heat source and sample exists in the heating and cooling progresses of most nucleic acid amplification system. The retard would result in the problem that the sample would take a long time to reach the set temperature and the problem would reduce the speed of integrate reaction. Non-specific products would be created in the process of amplification when the sample cannot reach the set temperature within a certainly time and the amplified efficiency would be reduced. A miniaturization nucleic acid amplification system heated by air was designed in this study according to the principle of air-heated nucleic acid amplification system and the characteristics of the PCR instrument Smart-cycler. The heat transfer process was analyzed and the heat transfer time was calculated. The actual temperature was measured in real time, and the temperature curves were fitted. The heating time was chosen by analysis results and data fitting and the air temperature was changed, while the sample temperature was recorded. The retard between sample and air was optimized by choosing the best curve of sample temperature. The temperature retard between sample and air was reduced sharply and the required time of integrate progress is shortened to 50%. We confirmed from the amplification experiment of Listeria monocytogenes that the improved system could complete 3 cycles within 4 minutes, and the amplification effect was good. The amplification speed and effect could be improved effectively by optimizing the delay between sample and air.
To study the significance of T-lymphocytes rDNA transcription activity in diagnosis, differential diagnosis, therapeutical effect and evaluation of treatment for colorectal carcinoma, 59 cases of colorectal carcinoma, 20 cases of colorectal inflammatory disease and 9 volunteers were choosen to detect the T-lymphocyte rDNA transcription activity of peripheral blood T-lymphocyte by cell culture and CMIAS008 image analysis system of Ag-NOR. Results: T-lymphocytes rDNA transcription activity was decreased obviously in colorectal inflammatory patients. Compared with control group, both group showed markedly statistical difference (Ｐ＜0.01). Tlymphocytes rDNA transcription activity increased gradually to normal groups after operation and chemical treatment for colorectal carcinoma patients; but it decreased for recurrent patients three years after operation. Conclusions: The detection of T-lymphocytes transcription activity can be used as a differential criterion for colorectal carcinoma and colorectal inflammatory disease, meanwhile it also can be used as a reference criterion for evaluation of treatment and supervision of tumor recurrence.
The oncogene ras p21 expression and DNA content in 46 cases of colorectal tumor were analysed quantitatively with flow cytometry and cyto-immunofluorescence staining technique. The results showed that the positive rate of ras p21 expression was 65.7% and the rate of DNA aneuploid was 74.3% in colorectal carcinomas. Ras p21 expression was higher in colorectal adenocarcinomas than that of the adenomas and normal mucosa. DNA ploid and proliferative index had some association with ras p21 expresssion. Detection of ras p21 expression and DNA content in tumors may be helpful in predicting the outcome of colorectal cancer patients.
Objective To introduce the characteristics of tetrahedral framework nucleic acids (tFNA), focusing on its application in the treatment of osteoarthritis (OA) and relationship with microRNA (miRNA), and prospect the application of tFNA in the treatment of OA and the new idea of constructing miR-tFNA functional complex to treat OA. Methods Recent studies were extensively reviewed to analyze the mechanism of tFNA and its relationship with OA and miRNA. Results tFNA, a new type of new carrier, can not only play an indirect role in the treatment of OA as a small molecular carrier with therapeutic effect, but also play a direct role through the regulation of chondrocytes. It can bind with the miRNA that can regulate OA. The therapeutic effect of constructing tFNA functional complex loaded with miRNA has been verified in various diseases, and tFNA has advantages compared with other vectors. Conclusion tFNA, a novel framework nucleic acid structure, plays an important role in the treatment of OA. Constructing miR-tFNA functional complex may be an innovative idea in the treatment of OA.
Objective To investigate the role of long chain non coding ribonucleic acid (LNcRNA) small nucleolar RNA host gene 14 (SNHG14) in regulating microribonucleic acid 223-3p (miR-223-3p) in alleviating sepsis associated lung injury in rats. Methods Sepsis rat models were established and the rats were randomly divided into SNHG14 upregulation group, SNHG14 upregulated control group, SNHG14 downregulation group, SNHG14 downregulation control group, miR-223-3p upregulation group, miR-223-3p upregulation control group, miR-223-3p downregulation group, miR-223-3p downregulation control group, and model group. Sham operation group was set up simutalously. All of them were administered through the tail vein. After 48 hours, the lung tissues was euthanized to detect the expressions of LncRNA SNHG14 and miR-223-3p. Tumor necrosis factor α (TNF- α), interleukin-1β (IL-1β), interleukin-18 (IL-18), lung wet weight/dry weight ratio (W/D) and the alveolar fluid clearance rate (AFC) were detected. Pathological changes in lung tissues were observed. Human NOD like receptor family protein 3 (NLPR3), cysteine-requiring aspartate protease 1 (caspase-1), and 1-aminocyclopropane-1-carboxylate synthase (ACS) genes and proteins expressions in lung tissues were detected. The dual luciferase reporter gene experiment was used to verify the targeted regulatory effect of LncRNA SNHG14 on miR-223-3p. Results Compared with the sham operation group, the model group showed increase in lung tissue LncRNA SNHG14 expression, serum inflammatory factor levels, W/D, pathological change quantification score, and lung tissue NLPR3, caspase-1 and ACS expressions (P<0.05), decrease in miR-223-3p expression and AFC (P<0.05). Compared with the model group and corresponding control groups, the SNHG14 upregulation group showed increase in LncRNA SNHG14 expression (P<0.05), and the SNHG14 upregulation group and the miR-223-3p downregulation group showed decrease in miR-223-3p expression and AFC (P<0.05), and increase in the levels of serum inflammatory factors, W/D, quantitative scores of pathological changes, and the expressions of NLPR3, caspase-1 and ACS in lung tissues (P<0.05). Compared with the model group and corresponding control groups, the expression of LncRNA SNHG14 in the SNHG14 downregulation group decreased (P<0.05), while the expression of miR-223-3p and AFC in the SNHG14 downregulation group and miR-223-3p upregulation group increased (P<0.05), which showed decrease in the levels of serum inflammatory factors, W/D, quantitative scores of pathological changes, and the expressions of NLPR3, caspase-1 and ACS in lung tissues (P<0.05). There were binding sites between LncRNA SNHG14 and miR-223-3p, and the former could negatively feedback targeted to regulate the latter. Conclusion Downregulation of LncRNA SNHG14 targets an increase in miR-223-3p expression and inhibit the NLRP3 pathway to alleviate sepsis related lung injury, which is related to the inhibition of inflammatory response, while upregulation of LncRNA SNHG14 negatively feedback targeting miR-223-3p and activated the NLRP3 pathway to exacerbate sepsis related lung injury.
ObjectiveTo study the expression and role of homeobox transcription antisense intergenic ribose nucleic acid (HOTAIR) in CD133-positive gastric cancer cells, which was classified to long non-coding RNA (LncRNA). MethodsImmune magnetic cell sorting (MACS) was performed to sort CD133-positive and CD133-negative cells of KATO-Ⅲgastric cancer cells, then reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the expressions of HOTAIR mRNA and CD133 mRNA. After the intervention of small interfering RNA (siRNA) for CD133-positive KATO-Ⅲcells, RT-PCR method was performed to detect the expression of HOTAIR mRNA to select siRNA who had the best silent effect. The selected-siHOTAIR was used to silent the expression of HOTAIR, then the expressions of CD133 mRNA, E-cadherin mRNA, and N-cadherin mRNA were detected by RT-PCR. At last, Transwell experiments were performed to detect the migration ability and invasion ability. Results①?RT-PCR test results showed that, the expression levels of CD133 mRNA and HOTAIR mRNA in CD133-positive group were significantly higher than those of CD133-negative group and no separation group (P < 0.05).②?After interference of siHOTAIR, the expression levels of HOTAIR mRNA in siHOTAIR1 group, siHOTAIR2 group, and siHOTAIR3 group were all significantly lower than those of blank control group and negative control group (P < 0.05), and the expression levels of HOTAIR mRNA in siHOTAIR2 group was lower than those of siHOTAIR1 group and siHOTAIR3 group (P < 0.05). The results indicated that siHOTAIR2 had the best interference efficiency.③?The expression levels of CD133 mRNA and N-cadherin mRNA in siHOTAIR2 group were lower than those corresponding indicators of blank control group and negative control group (P < 0.05), but the expression level of E-cadherin mRNA was higher than those of blank control group and negative control group (P < 0.05). Transwell experiment results showed that, number of cells which through the cell membrane in siHOTAIR2 group was lower than those of blank control group and negative control group (P < 0.05). ConclusionThe expression of HOTAIR mRNA in CD133-positive KATO-Ⅲgastric cancer cells was higher than that of CD133-negative cells, interfering the expression of HOTAIR mRNA can reduce the expression of CD133 mRNA in CD133-positive KATO-Ⅲgastric cancer cells, and can inhibit cell migration and invasion.
ObjectiveTo analyze the influencing factors for re-positive nucleic acid test in discharged coronavirus disease 2019 (COVID-19) patients in Chengdu, Sichuan Province, and to provide data support for the epidemics prevention and control. MethodsThe clinical data of 660 discharged COVID-19 patients from January 23, 2020 to February 28, 2021 in our center were retrospectively analyzed. The patients were divided into two groups according to the reexamination of virus nucleic acid, including a negative group [549 patients, including 428 males and 121 females with a median age of 33.0 (28.0, 48.0) years] and a positive group [111 patients, including 76 males and 35 females with a median age of 39.0 (28.0, 51.0) years]. The clinical data of the two groups were compared. Results The re-positive rate of the discharged patients was 16.82%. Univariate analysis showed that the re-positive rate of females was higher than that of males (χ2=4.608, P=0.032). The re-positive rate of confirmed patients was higher than that of asymptomatic infected patients (χ2=8.140, P=0.004). The re-positive rate of domestic patients was higher than that of imported patients (χ2=9.178, P=0.002). The counts of CD3+ (P=0.038), CD4+ (P=0.048) and CD8+ (P=0.040) T lymphocytes in the negative group were higher than those in the positive group. The binary logistic regression analysis showed that the clinical classification and CD8+ T lymphocyte count were independent risk factors affecting the recurrence of virility. ConclusionThe gender, origin, T lymphocyte subsets count and clinical type are the influencing factors for re-positive result, and clinical type and CD8+ T lymphocyte count are the independent influencing factors for re-positive result. Therefore, improving the immunity of infected patients, as well as early detection and timely treatment are effective means to reduce the re-positive occurrence.
As the COVID-19 pandemic is intensifying globally, more and more people are pinning their hopes on the development of vaccines. At present, there are many research teams who have adopted different vaccine technology routes to develop 2019-nCoV vaccines. This article reviews and analyzes the current development and research status of 2019-nCoV vaccines in different routes, and explores their possible development in the future.