【Abstract】ObjectiveTo measure the expressions of Fas/FasL mRNA in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma, and to explore the relationship between the expressions of Fas/FasL mRNA in those tissues and the hepatocellular carcinogenesis. MethodsSemi-quantity reverse transcript-ploymerase chain reaction(QRTPCR) were performed to measure the relative quantity of the Fas and FasL mRNA expressions in normal liver (n=25), adjacent noncancerous liver parenchyma(n=40) and hepatocarcinoma(n=40). ResultsThe relative quantity of Fas and FasL mRNA expressed in normal liver, adjacent non-cancerous liver parenchyma and hepatocarcinoma were 0.792±0.039 vs 0.245±0.043，0.857±0.031 vs 0.429±0.035 and 0.473±0.047 vs 0.185±0.041, respectively. The relative quantity of Fas mRNA expression in hepatocarcinoma was lower than that of normal liver tissue and adjacent non-cancerous liver parenchyrna (P＜0.05). The relative quantity of FasL mRNA expression in hepatocarcinoma was also lower than that of normal liver tissue (P＜0.05) and adjacent non-cancerous liver parenchyma (P＜0.01), but its expression in adjacent non-cancerous liver parenchyma was higher than that of normal liver tissue (P＜0.05).ConclusionHepatorcarcinoma may escape the immune surveillance of the host, not only by means of reducing Fas expression, but also through adjacent non-cancerous liver parenchyma’s increasing expression of FasL to induce apoptosis of contact lymphocyte which highly expresses Fas.
Objective To investigate the expression and clinical value of long chain non-coding RNA nicotinamide nucleotide hydrogenase antisense RNA1 (LncRNA NNT-AS1), motor neuron and pancreas homeobox protein 1 antisense RNA1 (MNX1-AS1) in lung cancer patients. Methods This study selected 128 patients diagnosed with lung cancer admitted to The Third Medical Center of the General Hospital of the People’s Liberation Army from April 2020 to April 2021 as a cancer group. During the same period, 128 patients with benign pulmonary nodules were regarded as a benign group, and 128 healthy individuals who underwent physical examination were selected as a control group. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied to detect the levels of LncRNA NNT-AS1 and MNX1-AS1 in serum. A three-year follow-up was conducted on all lung cancer patients, with 52 patients in the death group and 76 patients in the survival group. Receiver operator characteristic (ROC) curve was applied to analyze the diagnostic value of serum LncRNA NNT-AS1 and MNX1-AS1 for the occurrence of lung cancer and their predictive value for prognosis. Results Compared with the control group, the serum levels of LncRNA NNT-AS1 and MNX1-AS1 were obviously increased in the benign group and the cancer group (P<0.05). Compared with the benign group, the levels of LncRNA NNT-AS1 and MNX1-AS1 in serum of the cancer patients were obviously increased (P<0.05). The area under ROC curve (AUC) of serum LncRNA NNT-AS1 combined with MNX1-AS1 for the diagnosis of lung cancer was higher than that of LncRNA NNT-AS1 and MNX1-AS1 alone (ZLncRNA NNT-AS1~LncRNA NNT-AS1+MNX1-AS1=2.496, P=0.013; ZMNX1-AS1~LncRNA NNT-AS1+MNX1-AS1=2.831, P=0.007). The levels of LncRNA NNT-AS1 and MNX1-AS1 were related to tumor differentiation, clinical stage, and lymph node metastasis (P<0.05). Compared with the survival group, the serum levels of LncRNA NNT-AS1 and MNX1-AS1 in the death group were obviously increased (P<0.05). The AUC of combined prediction for lung cancer prognosis by serum LncRNA NNT-AS1 and MNX1-AS1 was higher than that predicted by LncRNA NNT-AS1 and MNX1-AS1 alone (ZLncRNA NNT-AS1~LncRNA NNT-AS1+MNX1-AS1=2.539, P=0.011; ZMNX1-AS1~LncRNA NNT-AS1+MNX1-AS1=3.377, P=0.001). Conclusion LncRNA NNT-AS1 and MNX1-AS1 are highly expressed in serum of lung cancer patients, and both have certain value in diagnosis and prognosis evaluation of lung cancer.
ObjectiveTo investigate the synergistic effect of cold stress plus particulate matter 2.5 (PM2.5) co-exposure on the occurrence of respiratory inflammation and the possible post-transcriptional regulation mechanism of cold inducible RNA-binding protein (CIRP).MethodsIn vivo and in vitro experiments were carried out, and the lung tissue specimens from human surgical resection were observed. The rat model and cultured airway epithelial cells 16HBE were respectively divided into four groups (n=8), namely blank control group, 5 °C/18 °C group, PM2.5 group and 5 °C/18 °C+PM2.5 group. The expression of mRNA and protein of representative inflammatory cytokines and CIRP of cultured airway epithelial cells and rat bronchial/pulmonary tissues were respectively detected by ELISA, qPCR, and Western blot. Furthermore, the temporal dynamics of CIRP distribution were observed by cellular immunofluorescence. Finally, immunohistochemical method was used to observe the localization and expression of CIRP in rat and human bronchial/pulmonary tissues at the same time.ResultsIn vivo experiments, the mRNA and protein expression levels of CIRP, interleukin-6, and tumor necrosis factor-α in 5 °C group and PM2.5 group were significantly higher than those in the control group (all P<0.05), while the expression level of mRNA and protein in 5 °C+PM2.5 group were increased most obviously (all P<0.01). The same rule also appeared in the experimental results of each group in the vitro experiment. In addition, CIRP was mainly located in the cell nucleus; compared with the control group, the intracellular shift of CIRP appeared in 18 °C group and PM2.5 group, while the migration phenomenon was most obvious in the 18 °C+PM2.5 group. In the immunohistochemistry of rat bronchus/pulmonary tissue, the expressions of CIRP in the 5 °C group and in the PM2.5 group were significantly higher than those in the control group, and the CIRP expression in 5 °C+PM2.5 group was increased most evidently. Moreover, CIRP was expressed in the bronchial epithelial mucosa of normal people and patients with chronic obstructive respiratory disease (COPD), and it is mainly located in the nucleus of airway mucosal epithelial cells. The CIRP expression of COPD patients was significantly higher than that in the normal population.ConclusionCold stress has a sensitizing effect on airway epithelial inflammatory response induced by PM2.5, and post-transcriptional regulation of CIRP translocation from nucleus to cytoplasm may be an important mechanism.
ObjectiveTo summarize the research progress of microRNA (miRNA) in the osteoarthritis (OA) cartilage matrix degradation. MethodsThe domestic and foreign related literature about the miRNA in the OA cartilage matrix degradation was reviewed, summarized, and analyzed. ResultsOA is a common chronic joint disease characterized by cartilage degeneration, its etiology and pathogenesis are still not completely clear. miRNA, a kind of small single stranded non-coding RNA molecule, is closely correlated with inflammatory mediators and various cytokines during the cartilage matrix degradation, suggesting that miRNAs have important regulatory functions at the molecule and cellular levels. ConclusionmiRNA can serve as potential biomarkers and will give new insight into diagnosis and therapeutic strategies in OA.
The occurrence and development of myopia is closely related to scleral remodeling. Therefore, in order to effectively prevent and cure myopia, it is very important to clarify the mechanism of scleral remodeling. In recent years, Chinese scholars have found that endoplasmic reticulum stress can regulate the expression of apoptotic proteins through the inositol demand protein-1/X box binding protein-1 pathway in the unfolded protein response, thus it is involved in regulating the state of scleral fibroblasts under hypoxia and regulating the occurrence and development of scleral remodeling. At the same time, some studies have found that inhibiting and knocking out protein kinase RNA-like endoplasmic reticulum kinase and activated transcription factor 6 in endoplasmic reticulum stress can effectively inhibit the growth of ocular axis. This proves that endoplasmic reticulum stress plays an important role in the occurrence and development of scleral remodeling. However, the comprehensive analysis of endoplasmic reticulum stress and scleral remodeling has not been reported at home and abroad. In-depth analysis of the relationship between endoplasmic reticulum and scleral remodeling is of great significance for the follow-up analysis and study of the mechanism of scleral remodeling.
Diabetic retinopathy (DR) is a serious complication of diabetes mellitus that not only impairs vision and quality of life but has also emerged as a leading cause of blindness in working-age individuals. Long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (LncMALAT1) is a non-coding RNA molecule that regulates gene expression and has been implicated in the pathogenesis and progression of DR. It exerts its effects through the modulation of various pathological processes, including inflammation, oxidative stress, angiogenesis, and apoptosis. Notably, alterations in the expression levels of LncMALAT1 may serve as potential biomarkers for the early diagnosis of DR. Furthermore, interventions targeting LncMALAT1, employing antioxidants, anti-angiogenic agents, traditional Chinese medicine, and gene therapy, present promising avenues for its potential development as an effective therapeutic target for DR.
The mechanisms behind diabetic retinopathy (DR) can be ascribed primarily to retinal microvascular abnormalities, excessive inflammatory response and neurodegeneration. Circular RNA (circRNA) is a type of endogenous non-coding RNA with a special circular structure, which is mainly composed of precursor RNA after shearing and processing. It is widely present in the retina and participates in the occurrence and development of various fundus diseases. CircRNAs express in an abnormal way in retina, serving as “the sponge” for miRNA so as to play roles in dysfunction of retinal vascular, inflammatory response and neurodegeneration in the development of DR. Further studies for circRNAs in DR will illustrate pathophysiology of DR more deeply, shedding light on circRNAs becoming novel biomarkers and molecular targets for diagnosis and treatment, thus achieving the goal of early diagnosis and precise therapy of DR.
Objective To construct vectors that express phosphatidylinositol-3-kinase, catalytic, beta polypeptide （PIK3cb） shRNA in eukaryon plasmid catalyzed by PI3K in rat, then test their effects on intimal hyperplasia in transplanted vein graft. Methods One hundred and fifty SD rats were randomly divided into six groups （n=25, in each group）: blank (25% Pluronic F-127), shRNA-1, shRNA-2, 1/2 （shRNA-1+shRNA-2）, negative control (pGenesil-1 scramble shRNA) and positive control （wortmannin） group. The jugular vein in rats were interpositioned autologously into the common carotid artery. shRNA and 25% Pluronic F-127 were mixed and coated around the transplanted vein in three PIK3cb shRNA groups. Every 5 samples were removed according to the time point (1, 3, 7, 14 and 28 days after operation), respectively. The thickness of intima and neointima area were calculated and analyzed by computer system. The PCNA expression was detected by Western blot and SP immunohistochemistry. Results The intimal thickness of three PIK3cb shRNA groups were lower than those in the blank group and negative control group on day 3, 7, 14, 28 after operation (P＜0.05); The neointima area in three PIK3cb shRNA groups (except shRNA-2 group on day 3, 7) began to decrease significantly from day one (P＜0.05). The protein expression of PCNA in three PIK3cb shRNA groups on day 3 after operation were decreased compared with blank group and negative group (P＜0.05). The percentage of the PCNA positive cells area in three PIK3cb shRNA groups were significantly lower than those in blank group and negative control group in each time point (Plt;0.05). There were no significant differences between blank and negative control group in different time points (Pgt;0.05). Conclusion The PIK3cb shRNA can effectively inhibit the proliferation of vascular smooth muscle cell, which may provide a new gene therapy for the prevention of vein graft restenosis after bypass grafting.
Objective To analyze the characteristics of intestinal flora in patients with allergic asthma, so as to provide a theoretical basis for the development of new clinical treatment methods. Methods Fecal samples were collected from 14 patients with allergic asthma and 15 healthy people between January 2021 and December 2021, and 16S rRNA was used to analyze the composition and diversity of intestinal flora of the participants. Results There was no statistically significant difference in age, gender, BMI, or smoking history between the allergic asthma group and the control group (all P>0.05). Alpha diversity results showed that there was significant difference in the abundance of intestinal flora between the two groups, but there was no significant difference in the diversity of intestinal flora between the two groups. The results of β diversity analysis indicated that there were significant differences in the composition of bacterial flora between the allergic asthma group and the control group. The difference bacteria between the two groups at the genus level are Faecalibacterium, Roseburia, Alistipes, Sphingomonas, Dorea, Ruminococcaceae_UCG-002, Streptomyces, [Eubacterium]_venturiosum_group, Butyriococcus and Agathobacter. Conclusion Compared with healthy individuals, patients with allergic asthma have undergone significant changes in the composition of their gut microbiota, with various differential bacteria present. Among them, Roseburia and Eubacterium may be involved in the pathogenesis of allergic asthma through changes in short chain fatty acids.
Objective To investigate the effects of knocking down Rac1 gene (ras-related C3 botulinum toxin substrate 1) by small hairpin RNA (shRNA) on retinal neovascularization in a mouse model of oxygen-induced retinopathy (OIR). Methods One hundred and eight 7-day-old C57BL/6J mice were divided into three groups randomly.The OIR was induced by Smith protocol in 2 groups. OIR mice received an intravitreal injection of Rac1-shRNA plasmid or the nonsense plasmid in the geneintervention group and control group respectively at the age of postnatal day 11 (P11). Non-OIR mice also received an intravitreal injection of Rac1-shRNA plasmid at P11 as the blankintervention group which lived in the normoxic environment.Retinal neovascularization was investigated on flat-mounts after fluorescence angiography at P15 and P17. Endothelial cell nuclei breaking through the internal limiting membrane were counted on pathological section at P17.The expression of Rac1 and NF-kappa;B p65 subunit was measured by immuohistochemistry, Western blot, real-time polymerase chain reaction (RT-PCR) and in situ hybridization. Results Compared with the blank-control group,the level of Rac1 mRNA in the gene-intervention group decreased obviously（t=4.500，P=0.001）;the retinal non-perfusion areas,fluorescence leakage, neovascularization and the number of endothelial cell nuclei breaking through the internal limiting membrane were reduced significantly（t=6.521,P＜0.001）; the level of NF-kappa;B p65 nuclear translocation decreased（t=16.008,P＜0.001）while the expression of NF-kappa;B p65 mRNA was reduced obviously（t=3.354,P=0.006）, which was positively correlated with the expression of Rac1-mRNA (P=0.012).Conclusion Intravitreal injection of Rac1-shRNA with liposome in mice can effectively inhibit the expression of Rac1,and inhibit the retinal neovascularization under relative hypoxia via blocking the ROS-NF-kappa;B pathway.