1.Association of intestinal microecology with the development and progression of autoimmune hepatitis
Yuyan XIA ; Qiongrong ZENG ; Fujian LI ; Fengyan LI ; Qi LI ; Lixia TANG ; Guo ZHANG
Journal of Clinical Hepatology 2024;40(2):374-379
		                        		
		                        			
		                        			Autoimmune hepatitis (AIH) is an autoimmune disease characterized by chronic liver inflammation, with a gradually increasing incidence rate, and its social and medical burdens cannot be neglected. Intestinal microecology is becoming a research hotspot in the field of autoimmune disease. In recent years, it has been believed that changes in intestinal microecology can cause changes in autoimmune state, microbial metabolites, and intestinal barrier, which is one of the driving factors for the onset of AIH. Early diagnosis and correct treatment can help to improve the prognosis of AIH patients. This article introduces the characteristics of gut microbiota in AIH patients, elaborates on the impact of intestinal microflora imbalance on the pathogenesis of AIH, and briefly describes related treatment regimens from the perspective of intestinal microecology, so as to comprehensively understand and explain the role of intestinal microecology in AIH and the impact of intestinal microecology balance on the pathogenesis, diagnosis, and treatment of AIH. 
		                        		
		                        		
		                        		
		                        	
2.Intervention Effect of Dabufei Decoction on Acute Lung Injury in Rats with High Altitude Hypoxia Based on HIF-1α/NLRP3 Signaling Pathway
Congyi LI ; Wangjie CAO ; Yong HUANG ; Yun SU ; Hongxia GONG ; Guangxian LENG ; Lixia LI ; Yuanding ZENG
Chinese Journal of Modern Applied Pharmacy 2024;41(6):736-742
		                        		
		                        			OBJECTIVE 
		                        			To investigate the intervention effect of Dabufei decoction on acute lung injury in rats with high altitude hypoxia by regulating the expression of the HIF-1α/NLRP3 signaling pathway and related molecules.
METHODS 
Sixty SPF SD rats were randomly divided into blank group, model group, positive drug group, Dabufei decoction high-dose, medium-dose, and low-dose groups with 10 rats in each group. After 3 d of adaptation to feeding, the rats in the blank group and model group were given the same amount of normal saline by gavage, and the rats in Dabufei decoction high-dose, medium-dose, and low-dose groups were given gavage for 14 d, respectively. The positive drug group was given dexamethasone by intraperitoneal injection for three consecutive days before entering the chamber. Except for the blank group, the rats in each group were exposed to hypoxia in the experimental animal low-pressure simulation chamber from the 15th day for three consecutive days. At the end of the experiment, the wet to dry ratio(W/D) of the rat lung tissue was detected. The morphology of lung tissue was observed by HE staining. ELISA detected the levels of IL-1β and IL-18 in serum. Western blotting and RT-qPCR were used to detect the protein and mRNA expressions of HIF-1α, NLRP3, GSDMD, and caspase-1 in the lung tissue of rats.
RESULTS 
The W/D value showed that compared with the blank group, the W/D of the model group was significantly increased(P<0.01). Compared with the model group, the W/D of rats in the positive drug group, Dabufei decoction high-dose group, medium-dose, and low-dose groups was significantly decreased(P<0.01 or P<0.05). HE results showed that compared with the blank group, alveolar septum thickening, pulmonary interstitial congestion, edema, inflammatory cell infiltration, and a small amount of exudation in the alveolar cavity were seen in the lung tissue of the model group. Compared with the model group, the thickening of alveolar walls in the positive drug group, Dabufei decoction high-dose group, medium-dose, and low-dose groups were reduced, and the pulmonary interstitial congestion, edema, and inflammatory cell infiltration were significantly reduced. ELISA results showed that IL-1β and IL-18 in rat serum were significantly higher in the model group than in the blank group(P<0.01). Compared with the model group, the levels of IL-1β and IL-18 in the serum of the rats in the positive drug group, Dabufei decoction high-dose group, medium-dose, and low-dose groups were significantly decreased(P<0.05 or P<0.01). Moreover, the results of Western blotting and RT-qPCR showed that compared with the blank group, the relative protein and mRNA expressions of HIF-1α, NLRP3, GSDMD, and caspase-1 in the lung tissue of the model group were significantly increased(P<0.01). Compared with the model group, the relative protein and mRNA of HIF-1α, NLRP3, caspase-1 and GSDMD in the positive drug group and Dabufei decoction high-dose group were significantly decreased(P<0.05 or P<0.01), the relative protein of HIF-1α, NLRP3, caspase-1 and GSDMD in Dabufei decoction medium-dose group were significantly decreased and HIF-1α, caspase-1 mRNA were significantly decreased(P<0.05), the relative protein of HIF-1α and GSDMD in the low-dose group was decreased(P<0.05). The positive drug group and Dabufei decoction high-dose group had the more significant effect.
CONCLUSION 
Dabufei decoction can regulate the HIF-1α/NLRP3 signaling pathway, inhibit pyroptosis and reduce inflammation, and has a certain protective effect on acute lung injury in rats with high altitude hypoxia.
		                        		
		                        		
		                        		
		                        	
3.Quality appraisal of hypoglycemia fear assessment tools for adults
Lixia CHEN ; Hui SHI ; Dezheng ZHU ; Ying ZENG
Chinese Journal of Nursing 2024;59(9):1072-1079
		                        		
		                        			
		                        			Objective To evaluate the quality of hypoglycemia fear assessment tools for adults,so as to provide references for the selection of high-quality research tools.Methods PubMed,Embase,Web of Science,CNKI,VIP,Wanfang,CBM were retrieved to collect the literature on the measurement properties of hypoglycemic fear.There were 2 researchers who screened the literature and extracted data,independently.Based on the consensus-based standards for the selection of health measurement instruments(COSMIN)guideline,the methodological and measure-ment properties quality of the included assessment tools were evaluated by the COSMIN risk of bias checklist and quality criteria.Results A total of 17 studies involving 6 hypoglycemia fear assessment tools for adults were included,none of which reported cross-cultural validity,measurement error,and responsiveness.All assessment tools were recommended by grade B.Conclusion The 15-item Fear of Hypoglycemia Scale(FH-15)has the most comprehensive measurement properties in its evaluation,and can be provisionally recommended for use,but it needs further testing.
		                        		
		                        		
		                        		
		                        	
4.The effect of Rhubarb ultrasonic electroconductive targeted dialysis therapy on the recovery of gastrointestinal dysfunction in sepsis patients:an observational study
Minggui CHEN ; Xuefei LIANG ; Fangfang WANG ; Lixia HUANG ; Ruixiang ZENG ; Yafeng HE ; Minzhou ZHANG ; Jiaping ZENG ; Xiaoxuan ZHANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2024;31(3):262-266
		                        		
		                        			
		                        			Objective To explore the effect of Rhubarb ultrasonic electrical conduction target penetration therapy on the gastrointestinal function of sepsis patients.Methods A total of 88 cases of inpatient sepsis patients diagnosed and treated at the Second Affiliated Hospital of Guangzhou University of Chinese Medicine from October 2021 to June 2023 were selected as the participants and the patients were divided into the treatment group and the control group by a simple random method.Both groups were given routine treatments such as anti-infection,fluid resuscitation,nutritional support,blood sugar management,and prevention of stress ulcers,while the treatment group was treated with ultrasonic conductance target penetration Rhubarb therapy at Shenque point and Zhongwan point on the basis of the control group.The main observational indexes were the number of bowel sounds in the two groups before treatment and after 3 days,and 7 days of treatment.The secondary observational indexes were gastric retention,serum C-reactive protein(CRP),procalcitonin(PCT),acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ),the first defecation time,time to mechanical ventilation,length of stay in the intensive care unit(ICU),hospitalization cost,and 28 days readmission rate of the two groups.The gastric retention was evaluated and recorded every 4 hours before treatment and 3 days and 7 days after treatment,CRP and APACHE Ⅱ score were evaluated before treatment and 7 days after treatment,the 28-day readmission was followed up by telephone.Results A total of 44 cases in the treatment group and 43 cases in the control group completed the intervention.The difference between the bowel sounds and the amount of gastric retention of the two groups was not statistically significant before treatment,with the prolongation of time,the bowel sounds of the two groups gradually increased and the amount of gastric retention gradually decreased,and the number of bowel sounds in the treatment group increased significantly compared to the control group 3 days and 7 days after treatment[beats/minute:4.49(4.23,4.74)vs.3.07(2.80,3.33)3rd day of and 4.79(4.49,5.10)vs.3.36(3.06,3.66)on the 7th day of treatment,both P<0.01],and the amount of gastric retention was significantly reduced compared with the control group[mL:20.93(9.22,32.64)vs.53.52(41.16,65.88)for 3rd day of and 3.72(0.17,7.28)vs.31.59(24.87,38.31)on the 7th day of treatment,both P<0.01].The first defecation time was significantly shorter in the treatment group than in the control group(hours:25.67±9.99 vs.33.64±11.30,P<0.01).After 7 days treatment,the CRP,PCT,and APACHE Ⅱ score were significantly reduced in the treatment group compared to those in the control group[CRP(mg/L):45.97(35.68,56.26)vs.77.30(57.20,97.38),PCT(μg/L):0.94(0.56,1.31)vs.2.73(1.36,4.09),APACHE Ⅱ score:24.11±7.01 vs.28.06±9.25,all P<0.05],the mechanical ventilation time and ICU length of stay were significantly shortened[mechanical ventilation time(hours):107.05±70.76 vs.168.83±136.62,ICU stay time(days):7.58±3.72 vs.9.70±5.15,both P<0.05],the hospitalization cost and 28-day readmission rate were significantly reduced[hospitalization cost(yuan):80337.89±36483.72 vs.109100.24±87080.84,28-day readmission rate:9.1%(4/44)vs.27.9%(12/43),both P<0.05].Conclusion Rhubarb ultrasonic conductive target penetration therapy is a safe,simple,and effective characteristic therapy of external Chinese medicine that can provide a new basis for the protection of gastrointestinal function in patients with sepsis.
		                        		
		                        		
		                        		
		                        	
5.Correlation analysis between early visual function impairment and neuron function abnormality in diabetic mouse primary visual cortex
Haoqiong TANG ; Lixia FENG ; Tianchang TAO ; Yige ZENG
Chinese Journal of Experimental Ophthalmology 2024;42(8):698-704
		                        		
		                        			
		                        			Objective:To investigate the damage of visual function in the early stage of diabetes mellitus and the response characteristics of primary visual cortex neurons in diabetic mice.Methods:Twenty 7-week-old SPF grade male C57BL/6J mice were randomly divided into diabetes group and normal control group by the random number table method, with 10 mice in each group.The diabetes model in diabetes group was established by the intraperitoneal injection of streptozotocin.Fasting blood glucose concentration and body mass of mice were measured before and 1, 2 and 3 weeks after modeling, and intraperitoneal glucose tolerance test was performed 4 weeks after modeling to evaluate the establishment of diabetes model.At 4 weeks after modeling, the electroretinogram (ERG) responses of mice were recorded at dark adaptation luminances of 0.01 and 3.0 cd·s/m 2, and the ERG response to light adaptation luminance of 3.0 cd·s/m 2 was recorded 10 minutes after light adaptation to evaluate the retinal function of mice.The fundus of mice was examined with an ultra-wide field laser scanning ophthalmoscope.The visual function of mice was evaluated via the cutoff frequency of grating discrimination detected by visual water maze.The spatial frequency tuning curves of primary visual cortex (V1) neurons were detected by in vivo electrophysiology technique and the maximum firing intensity, self-firing intensity, optimal spatial frequency, cutoff frequency and bandwidth of neurons were calculated to evaluate the neuronal function of mice.The research scheme was approved by the Experimental Animal Ethics Committee of Anhui Medical University (No.LLSC20230419). Results:The diabetic model was successfully established in 10 mice in the diabetes group.At 4 weeks after modeling, compared with the normal control group, the b-wave amplitudes of mice at a dark adaptation luminance of 0.01 cd·s/m 2, the a- and b-wave amplitudes of mice at a dark adaptation luminance of 3.0 cd·s/m 2 and the b-wave amplitudes of mice at a light adaptation luminance of 3.0 cd·s/m 2 after 10 minutes of light adaptation showed a downward trend in diabetes group, but the differences were not statistically significant (all at P>0.05).Ultra-wide field laser scanning ophthalmoscopy showed no obvious vascular changes in the retina of diabetic mice.The results of visual water maze detection showed that the cutoff frequency of diabetes group was (0.45±0.06)c/d, which was significantly lower than (0.58±0.05)c/d of normal control group ( t=5.10, P<0.05). In vivo electrophysiological results showed that the maximum firing intensity of neurons in V1 region in diabetes group 4.29(2.60, 8.33)spikes/second, which was significantly lower than 7.10(4.34, 11.6)spikes/second in normal control group( Z=-4.29, P<0.05).The optimal spatial frequency, cutoff spatial frequency and bandwidth were 0.03(0.02, 0.05), 0.07(0.05, 0.12) and 0.14(0.07, 0.22)c/d in diabetes group, which were significantly lower than 0.41(0.03, 0.05) and 0.10(0.07, 0.14), 0.14(0.10, 0.26)c/d of normal control group, and the differences were statistically significant ( Z=-3.22, -3.19, -2.19; all at P<0.05). Conclusions:The abnormal visual function may occur in the early stage of diabetes before the appearance of retinal vasculopathy, which is related to the damage of neurons in the V1 region.
		                        		
		                        		
		                        		
		                        	
6.Protective Effect of Huangqi Baihe Granules on Acute Brain Injury in Rats with High Altitude Hypoxia Based on HIF-1α/NF-κB/NLRP3 Signal Pathway
Yuanding ZENG ; Yun SU ; Hongxia GONG ; Wangjie CAO ; Yongqi LIU ; Yong HUANG ; Guangxian LENG ; Lixia LI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):134-141
		                        		
		                        			
		                        			ObjectiveTo observe the effect of Huangqi Baihe granules on the hypoxia-inducible factor 1α (HIF-1α)/nuclear factor-κB (NF-κB)/NOD-like receptor hot protein domain related protein 3 (NLRP3) signaling pathway in a rat model of high altitude hypoxia. MethodSixty male SPF SD rats were randomly divided into blank group, model group, dexamethasone group (5 mg·kg-1), and high, middle, and low-dose groups of Huangqi Baihe granules (4.1, 2.05, 1.025 g·kg-1). Among them, each Chinese medicine group was administrated orally for continuously 14 d, once a day, and the dexamethasone group was injected intraperitoneally for continuously 3 d as the positive control group. On the 15th d, the model group, dexamethasone group, and high, middle, and low dose groups of Huangqi Baihe granules were exposed to the simulated high altitude, low pressure, and low oxygen environment in the animal low-pressure simulation cabin, and the exposure lasted for 3 d. Blood was collected from the abdominal aorta and serum was separated, and the brain tissue was taken after being killed. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in brain tissue. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in rat serum. Western blot was used to detect HIF-1α, NLRP3, phosphorylated nuclear factor-κB (p-NF-κB), NF-κB, desquamation D (GSDMD), and cysteine aspartate-specitis protein-1(Caspase-1) in rats of each group. The mRNA expression levels of HIF-1α, NLRP3, NF-κB p65, GSDMD, and Caspase-1 were detected by real-time quantitative polymerase chain reaction (Real-time PCR). ResultThe results of HE staining showed that as compared with the normal group, the pathological sections of brain tissues in the model group showed that pyramidal cells were loosely arranged and distributed in disorder, with different sizes. Compared with the model group, the pathological changes in pyramidal cells in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules were reduced. The results of ELISA showed that as compared with the normal group, the content of TNF-α, IL-6, and IL-1β in the serum of rats in the model group was significantly higher (P<0.01). Compared with the model group, the content of TNF-α, IL-6, and IL-1β in the serum of rats in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules decreased significantly (P<0.05, P<0.01). The results of Western blot showed that as compared with the normal group, the relative protein expression levels of HIF-1α, NLRP3, p-NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of the model group were significantly higher (P<0.01). As compared with the model group, the relative expressions of HIF-1α, NLRP3, p-NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of rats in the dexamethasone group and the high-dose group of Huangqi Baihe granules were significantly decreased (P<0.05, P<0.01). The relative protein expression levels of HIF-1α, NLRP3, p-NF-κB p65, and Caspase-1 in the brain tissue of rats in the middle-dose group of Huangqi Baihe granules decreased significantly (P<0.01), and the relative protein expression of HIF-1α in the brain tissue of rats in the low-dose group of Huangqi Baihe granules was reduced (P<0.05). The Real-time PCR analysis showed that as compared with the normal group, the mRNA expression levels of HIF-1α, NLRP3, NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of the model group were significantly increased (P<0.01). As compared with the model group, the mRNA expression levels of HIF-1α, NLRP3, NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of rats in the dexamethasone group were significantly decreased (P<0.01). The mRNA expression levels of HIF-1α, NF-κB p65, GSDMD, and Caspase-1 in the brain tissue of rats in the high-dose group of Huangqi Baihe granules decreased significantly (P<0.01). The mRNA expression levels of HIF-1α, NLRP3, and Caspase-1in the brain tissue of rats in the middle-dose group of Huangqi granules decreased (P<0.05, P<0.01). ConclusionThe protective effect of Huangqi Baihe granules on acute brain injury in low-pressure hypoxic rats may be related to the HIF-1α/NF-κB/NLRP3 signaling pathway. 
		                        		
		                        		
		                        		
		                        	
7.Application of outcome-based education combined with team-based learning in practice teaching of pediatric emergency and critical care nursing
Xue ZENG ; Xiaoyu ZHU ; Dan XU ; Lixia XIE ; Junjie ZHANG
Chinese Journal of Medical Education Research 2023;22(8):1272-1275
		                        		
		                        			
		                        			Objective:To investigate the application value of outcome-based education (OBE) combined with team-based learning (TBL) in the practice teaching of pediatric emergency and critical care nursing.Methods:A total of 84 nursing students who studied in the pediatric intensive care unit (PICU) of The First Affiliated Hospital of Air Force Medical University were selected and divided into control group and observation group. The 41 nursing students in the control group received traditional teaching, and the 43 nursing students in the observation group received OBE+TBL teaching. The two groups were assessed in terms of theoretical knowledge, practical operation ability, clinical thinking ability, and self-learning ability after teaching, and the degrees of satisfaction with teaching and participation in teaching were compared between the two groups. SPSS 22.0 was used for the t-test and the chi-square test. Results:After teaching, the observation group had significantly better theoretical knowledge, practical operation ability, and self-learning ability than the control group ( P<0.05). Compared with the control group, the observation group had significantly higher scores of learning engagement (recognition, behavior, emotion) ( P<0.05) and satisfaction with classroom effect, knowledge mastery, and learning interest ( P<0.05). Conclusion:The application of OBE+TBL teaching in PICU nursing students can effectively improve their self-learning ability and participation and help them to understand PICU nursing priorities and improve their practical operation ability.
		                        		
		                        		
		                        		
		                        	
8.Role of IFNLR1 gene in PRRSV infection of PAM cells
Ming QIN ; Wei CHEN ; Zhixin LI ; Lixue WANG ; Lixia MA ; Jinhong GENG ; Yu ZHANG ; Jing ZHAO ; Yongqing ZENG
Journal of Veterinary Science 2021;22(3):e39-
		                        		
		                        			Background:
		                        			Interferon lambda receptor 1 (IFNLR1) is a type II cytokine receptor that clings to interleukins IL-28A, IL29B, and IL-29 referred to as type III IFNs (IFN-λs). IFN-λs act through the JAK-STAT signaling pathway to exert antiviral effects related to preventing and curing an infection. Although the immune function of IFN-λs in virus invasion has been described, the molecular mechanism of IFNLR1 in that process is unclear. 
		                        		
		                        			Objectives:
		                        			The purpose of this study was to elucidate the role of IFNLR1 in the pathogenesis and treatment of porcine reproductive and respiratory syndrome virus (PRRSV). 
		                        		
		                        			Methods:
		                        			The effects of IFNLR1 on the proliferation of porcine alveolar macrophages (PAMs) during PRRSV infection were investigated using interference and overexpression methods. 
		                        		
		                        			Results:
		                        			In this study, the expressions of the IFNLR1 gene in the liver, large intestine, small intestine, kidney, and lung tissues of Dapulian pigs were significantly higher than those in Landrace pigs. It was determined that porcine IFNLR1 overexpression suppresses PRRSV replication. The qRT-PCR results revealed that overexpression of IFNLR1 upregulated antiviral and IFN-stimulated genes. IFNLR1 overexpression inhibits the proliferation of PAMs and upregulation of p-STAT1. By contrast, knockdown of IFNLR1 expression promotes PAMs proliferation. The G0/G1 phase proportion in IFNLR1-overexpressing cells increased, and the opposite change was observed in IFNLR1-underexpressing cells. After inhibition of the JAK/STAT signaling pathway, the G2/M phase proportion in the IFNLR1-overexpressing cells showed a significant increasing trend. In conclusion, overexpression of IFNLR1 induces activation of the JAK/STAT pathway, thereby inhibiting the proliferation of PAMs infected with PRRSV. 
		                        		
		                        			Conclusion
		                        			Expression of the IFNLR1 gene has an important regulatory role in PRRSVinfected PAMs, indicating it has potential as a molecular target in developing a new strategy for the treatment of PRRSV.
		                        		
		                        		
		                        		
		                        	
9.Role of IFNLR1 gene in PRRSV infection of PAM cells
Ming QIN ; Wei CHEN ; Zhixin LI ; Lixue WANG ; Lixia MA ; Jinhong GENG ; Yu ZHANG ; Jing ZHAO ; Yongqing ZENG
Journal of Veterinary Science 2021;22(3):e39-
		                        		
		                        			Background:
		                        			Interferon lambda receptor 1 (IFNLR1) is a type II cytokine receptor that clings to interleukins IL-28A, IL29B, and IL-29 referred to as type III IFNs (IFN-λs). IFN-λs act through the JAK-STAT signaling pathway to exert antiviral effects related to preventing and curing an infection. Although the immune function of IFN-λs in virus invasion has been described, the molecular mechanism of IFNLR1 in that process is unclear. 
		                        		
		                        			Objectives:
		                        			The purpose of this study was to elucidate the role of IFNLR1 in the pathogenesis and treatment of porcine reproductive and respiratory syndrome virus (PRRSV). 
		                        		
		                        			Methods:
		                        			The effects of IFNLR1 on the proliferation of porcine alveolar macrophages (PAMs) during PRRSV infection were investigated using interference and overexpression methods. 
		                        		
		                        			Results:
		                        			In this study, the expressions of the IFNLR1 gene in the liver, large intestine, small intestine, kidney, and lung tissues of Dapulian pigs were significantly higher than those in Landrace pigs. It was determined that porcine IFNLR1 overexpression suppresses PRRSV replication. The qRT-PCR results revealed that overexpression of IFNLR1 upregulated antiviral and IFN-stimulated genes. IFNLR1 overexpression inhibits the proliferation of PAMs and upregulation of p-STAT1. By contrast, knockdown of IFNLR1 expression promotes PAMs proliferation. The G0/G1 phase proportion in IFNLR1-overexpressing cells increased, and the opposite change was observed in IFNLR1-underexpressing cells. After inhibition of the JAK/STAT signaling pathway, the G2/M phase proportion in the IFNLR1-overexpressing cells showed a significant increasing trend. In conclusion, overexpression of IFNLR1 induces activation of the JAK/STAT pathway, thereby inhibiting the proliferation of PAMs infected with PRRSV. 
		                        		
		                        			Conclusion
		                        			Expression of the IFNLR1 gene has an important regulatory role in PRRSVinfected PAMs, indicating it has potential as a molecular target in developing a new strategy for the treatment of PRRSV.
		                        		
		                        		
		                        		
		                        	
10.Construction of miR-331-3p overexpression vector and its effect on cell proliferation.
Tao CHEN ; Lixia MA ; Jingxiang CUI ; Jinhong GENG ; Yongqing ZENG ; Wei CHEN
Chinese Journal of Biotechnology 2019;35(5):892-900
		                        		
		                        			
		                        			To investigate the effect of miR-331-3p on the proliferation of porcine renal epithelial cells (PK15) and its mechanism, the pcDNA 3.1(+) overexpression vector of miRNA-331-3p (pcDNA 3.1(+)-miR-331-3p) was constructed. PK15 cells were divided into four groups, including experimental group, experimental control group, inhibitor group and inhibitor control group. Experimental group and experimental control group were transfected with pcDNA 3.1(+)-miR-331-3p and pcDNA 3.1(+), respectively. Inhibitor group and inhibitor control group were transfected with miR-331-3p inhibitor and miR-331-3p negative control (miR-331-3p NC), respectively. Above all, CCK-8 reagent was used to plot the cell proliferation curve and Propidium (PI) staining was used to detect the proportion of cell stages. Secondly, its expression change were detected by quantitative real-time PCR that included the growth inhibitory protein family member 5 (ING5), cyclin-dependent kinase 2 (CDK2), cyclin-dependent kinase 3 (CDK3), cyclin-dependent kinase 4 (CDK4), Cyclin B and cyclin-dependent kinase inhibitor 1A (CDKN1A). The results showed that the expression of miRNA-331-3p was significantly increased in the experimental group. The cell proliferation curve showed that the number of cells in experimental group was significantly higher than that in experimental control group or inhibitor control group at 48 h and 72 h (P<0.05). Simultaneously, Inhibitor group was significantly lower than experimental control group or inhibitor control group in the number of cells at 48 h and 72 h (P<0.05), but there was no significant difference between the experimental group and the control group. Compared with the experimental control group, the proportion of cells of experimental group in G0/G1 phase decreased, the proportion of S phase and G2/M phase increased, and the inhibitor control group showed the opposite trend. Simultaneously, the expression levels of CDK2, CDK3, CDK4 and Cyclin B genes in the experimental group were significantly increased, while ING5 and CDKN1A genes inhibiting proliferation showed a significant downward trend. These results demonstrate that the miR-331-3p overexpression vector was successfully constructed, and miR-331-3p has the ability to promote the proliferation of PK15 cells. The study lays a solid foundation for further research for its role in pig growth and development.
		                        		
		                        		
		                        		
		                        			Animals
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		                        			Cell Line
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		                        			Cell Proliferation
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		                        			genetics
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		                        			Epithelial Cells
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		                        			cytology
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		                        			Genetic Vectors
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		                        			MicroRNAs
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		                        			genetics
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		                        			Swine
		                        			
		                        		
		                        	
            

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