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Objective To explore the value of dynamic nomogram constructed by multi spiral computed tomography(MSCT)features combined with inflammatory indicators in predicting the status of microvascular invasion(MVI)of hepatocellular carcinoma(HCC)before surgery.Methods The clinical and imaging data of 137 patients with postoperative pathologically confirmed HCC were analyzed retrospectively.According to the status of the MVI,they were divided into positive group(44 cases)and negative group(93 cases).Multivariate logistic regression analysis was used to screen independent risk factors for predicting the MVI status of HCC patients,and a joint prediction model was constructed,which was displayed in the form of a dynamic nomogram.The receiver operating characteristic(ROC)curve,calibration curve and Hosmer-Lemeshow test were used to evaluate the diagnostic efficiency,calibration and goodness of fit of the model,Akaike information criterion(AIC)and Bayesian information criterion(BIC)were used for comparison between the models,and a 5-fold cross-validation and decision curve analysis(DCA)were also used to evaluate the stability and clinical applicability of the model.Results Multivariate logistic regression analysis showed that necrosis and delayed-phase enhancement(DEd),and alkaline phosphatase to lymphocyte ratio(ALR)were independent risk factors for predicting MVI status in HCC patients.The area under the curve(AUC)of the dynamic nomogram was 0.721,with the sensitivity of 0.705 and the specificity of 0.656.The AIC and BIC values were 152.372 and 158.212,respectively.The calibration curve and the Hosmer-Lemeshow test showed that the model had a high degree of calibration and goodness of fit(χ2=2.372,P=0.967),the average AUC of the 5-fold cross-validation was 0.787,and the DCA showed that the nomogram model had a good clinical applicability.Conclusion The dynamic nomogram model constructed by MSCT features combined with inflammatory indicators is feasible to predict the MVI status of HCC patients before surgery,and the dynamic nomogram can directly generate the prediction results of different individuals.
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To explore the barriers and facilitators of shared decision making, 23 clinicians were selected for semi-structured interviews by purpose sampling and convenience sampling with phenomenological methods in qualitative research, and 7-step of Colaizzi was used to analyze the interview data. Three themes and twelve subthemes were extracted, included: individual factors of doctors (role cognition, perceived outcomes, communication skills, clinical expertise, cognitive bias) , individual factors of patients (general information, lack of disease knowledge, willingness to participate in decision making) and environmental factors (clinical situation, social environment, resources and social influence) . There were many barriers and facilitators in the implementation of doctor-patient shared decision making. It is necessary to scientifically analyze and actively deal with the influence of each factor, and find reasonable countermeasures to promote the clinical implementation of shared decision making.
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Objective To investigate the relationship between polymorphism of resistin(RETN)gene and metabolic associated fatty liver disease(MAFLD)in type 2 diabetes mellitus(T2DM)patients in middle and high altitude areas.Methods A total of 400 patients with T2DM in Qinghai area were recruited and divided into simple T2DM group(T2DM,n=200)and T2DM combined with MAFLD group(T2DM+ MAFLD,n=200)according to liver ultrasonography.Healthy individuals confirmed by physical examination were selected as the normal control group(NC,n=180).Plasma resistin levels were measured by ELISA.The polymorphism of RETN-420C/G and +299G/A genes were detected by PCR sequencing.Results By comparing the polymorphism of RETN-420C/G gene in each group,it was found that the frequencies of G/G genotype and G allele frequency in T2DM+MAFLD group were higher than those in NC group and T2DM group(P<0.05),while the frequencies of C/C genotype and C allele frequency were lower than those in NC group and T2DM group(P<0.05).The risk of MAFLD increased by 1.571,2.126 and 1.537 times respectively in T2DM patients with C/G,G/G genotype and G allele.Logistic regression analysis showed that G/G genotype was a risk factor for MAFLD in T2DM patients.By comparing the polymorphism of RETN+299G/A gene in each group,it was found that A allele frequency in T2DM+MAFLD group was higher than that in NC group and T2DM group,while G allele frequency was lower than that in NC group and T2DM group(P<0.05).The allele A increased the risk of MAFLD in T2DM patients by 1.432 times compared to allele G.Conclusion RETN gene-420C/G locus G/G genotype increases the risk of T2DM combined with MAFLD in middle and high altitudeareas.
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Objective To develop an evidence-based,localized practice protocol for the interhospital transfer of critically ill children.Methods Through a comprehensive evidence summary and semi-structured interviews,a preliminary inter-hospital transfer practice protocol for critically ill children was formulated.A panel of 31 experts from 12 hospitals in China participated in 2 rounds of expert correspondence between May and July 2022,facilitating meticulous revision of the protocol entries.Results The response rate for both rounds of questionnaires was 100%,and the expert authority coefficients ranged from 0.926 to 0.931.In the second round of consultation,the coefficient of variation for the importance score of each entry ranged from 0.036 to 0.226,and the Kendall's W was determined to be 0.201(P＜0.001).Additionally,the coefficient of variation for the feasibility score of each entry fell within the range of 0.070 to 0.314,with Kendall's W of 0.124(P＜0.001).Ultimately,the final interhospital transfer protocol for critically ill children comprised 8 level Ⅰ entries,16 level Ⅱ entries,and 75 level Ⅲ entries.Conclusion The interhospital transfer protocol constructed in this study is grounded in scientific evidence and exhibits practical feasibility.It serves as a valuable reference for organizing and implementing interhospital transfers of critically ill children.
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Objective To investigate the effect of acupuncture combined with Shenfu injection on brain injury and hemodynamics after cardiopulmonary resuscitation in patients with cardiac arrest.Methods A total of 100 patients who underwent cardiopulmonary resuscitation after cardiac arrest admitted to 903rd Hospital of the Joint Support Force of PLA from May 2019 to May 2022 were selected as the research objects,and they were divided into acupuncture group and combined group,with 50 cases in each group according to the random number table method.The acupuncture group was given acupuncture therapy,and the combined group was injected Shenfu injection on the basis of acupuncture group.S100β,neuron specific enolase(NSE),National Institutes of Health stroke scale(NIHSS)score,mini mental status examination(MMSE)score,hemodynamic index,Glasgow coma scale(GCS)score,acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)score and incidence of adverse reactions were compared between two groups.Results After treatment,S100β,NSE,NIHSS score,APACHE Ⅱscore,and heart rate(HR)of two groups were significantly lower than those before treatment,MMSE score,GCS score,cardiac output(CO),central venous pressure(CVP),mean arterial pressure(MAP)were significantly higher than those before treatment(P<0.05).S100β,NSE,NIHSS score,APACHE Ⅱ score and HR in combined group were significantly lower than those in acupuncture group,and MMSE score,GCS score,CO,CVP,and MAP were significantly higher than those in acupuncture group(P<0.05).The incidence of adverse reactions in combined group was significantly lower than that in acupuncture group(χ2=6.453,P=0.011).Conclusion Acupuncture combined with Shenfu injection treatment can improve brain injury and hemodynamics after cardiopulmonary resuscitation in patients with cardiac arrest,and the effect is significant.
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Objective To explore heterogeneous subtypes of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation and the characteristics differences of different types of children after liver transplantation. Methods Seven hundred and forty-one children who underwent living-related liver transplantation were enrolled. The self-designed general information questionnaire, Chinese version of 5-Item World Health Organization Well-Being Index (WHO-5) and the parent-report version of the Strengths and Difficulties Questionnaire (SDQ) were filled out by their guardians. The scores of five dimensions of SDQ were used as the manifest variables of the model. The classification model of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation was constructed by latent profile analysis. The latent categories of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation were analyzed. The influencing factors of latent categories were analyzed by univariate analysis and logistic regression model. Results There were three latent categories of psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation, including peer communication problem group (n=302), psychological and behavioral adaptation group (n=145) and psychological and behavioral adjustment difficulty group (n=294). The first two groups were merged into the psychological and behavioral health group (n=447), which had significant differences in the five dimensions and the total score of difficulties of SDQ compared with the psychological and behavioral adjustment difficulty group (n=294) (all P<0.001). Logistic regression analysis showed that age≤5 years old, primary disease of non-cholestatic liver disease, stem family were the risk factors for psychological and behavioral adjustment difficulties in pediatric recipients after liver transplantation. Female gender, high education levels of parents and high WHO-5 score of guardians were the protective factors for psychological and behavioral adjustment difficulties in pediatric recipients after liver transplantation (all P<0.05). Conclusions The psychological and behavioral adaptation characteristics of pediatric recipients after liver transplantation are heterogeneous. Medical staff should pay extensive attention to different characteristics of pediatric recipients after liver transplantation with different psychological and behavioral adaptation categories and adopt targeted screening and intervention strategies, aiming to improve psychological and behavioral adaptation outcomes of pediatric recipients after liver transplantation.
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To explore the barriers and facilitators of shared decision making, 23 clinicians were selected for semi-structured interviews by purpose sampling and convenience sampling with phenomenological methods in qualitative research, and 7-step of Colaizzi was used to analyze the interview data. Three themes and twelve subthemes were extracted, included: individual factors of doctors (role cognition, perceived outcomes, communication skills, clinical expertise, cognitive bias) , individual factors of patients (general information, lack of disease knowledge, willingness to participate in decision making) and environmental factors (clinical situation, social environment, resources and social influence) . There were many barriers and facilitators in the implementation of doctor-patient shared decision making. It is necessary to scientifically analyze and actively deal with the influence of each factor, and find reasonable countermeasures to promote the clinical implementation of shared decision making.
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Objective:To analyze the clinical features and genetic factors of neonatal 17β-hydroxysteroid dehydrogenase type10 (HSD10) deficiency.Methods:The clinical characteristics and genetic test results of a child with HSD10 deficiency coming from Children′s Hospital of Nanjing Medical University in April 2019 were retrospectively analyzed.The keywords" 17β-hydroxysteroid dehydrogenase type 10 deficiency" or " 2-Methyl3-Hydroxybutyryl-CoA dehydrogenase deficiency" or " HSD10" , etc.were searched in various databases, including CNKI, Wanfang, Weipu, Embase and PubMed to review the cases collected from all published data until May 31, 2020.Results:The patient was a newborn male who developed symptoms on the first day after birth.The main signs were metabolic acidosis, increased blood ammonia and lactate, and hypotonia.Trio whole exom sequencing in the patient and his parents identified hemizygous NM001037811: c.650G>A, p.R217Q in the HSD17B10 gene that is inherited from the mother.Since the child died on the third day after birth, no further central nervous system examination was performed.The mother of the child has intellectual disability, the sibling sister is normal and the HSD17B10 locus is wild type.By lite-rature reviewing, 5 newborn cases with clear medical records and genetic test results were listed.All patients were male, and had onset of HSD10 deficiency within 1 week after birth.The main phenotypes include metabolic acidosis (increased blood ammonia and lactate), hypoglycemia, hypotonia, and convulsions.All 6 children died in early infancy.The corresponsive HSD17B10 variants were c. 740A>G/p.N247S, c.677G>A/p.R226Q, c.257A>G/p.D86G and c. 650G>A/p.R217Q, which did not indicate the hot spots of mutation. Conclusions:HSD10 deficiency in the neonatal period is relatively rare.The clinical diagnosis is difficult due to the serious condition and short course of the disease.Severe metabolic acidosis, hypotonia, and convulsions in neonatal patients are the main reasons for the poor prognosis, which can be attributed to the hemizygous variation and heterogeneity of the mutation site in male patients.c.650G>A may be closely associated with severe neonatal HSD10 deficiency, but the molecular biological mechanism needs to be further clarified.HSD10 deficiency has a poor prognosis and lacks effective treatment.
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Objective:To investigate the clinical and pathogenic characteristics of community acquired pyogenic liver abscess (PLA).Methods:The clinical data of 172 patients in First Affiliated Hospital of Soochow University with community acquired PLA from March 2013 to September 2018 were retrospectively collected, including clinical characteristics, distribution of the causative pathogens, treatment regimens and outcomes. Chi-square test was used for statistical analysis.Results:There were 158(91.9%) cases with fever, 69(40.1%) cases with abdominal pain among 172 PLA cases. One hundred and forty-three (83.1%) were solitary, and 141(82.0%) cases localized in right hepatic lobe. One hundred and six (61.6%) cases were PLA of cryptogenic origin. There were 156 cases underwent etiology detection, with the positive etiology detection of 99(63.5%) cases. Ninety-two (92.9%) cases were infected with a single strain, and seven (7.1%) cases were infected with mixed strains. A total of 115 strains of bacteria were isolated. The main strains included 71 (61.7%) Klebsiella pneumoniae (KP), 21 (18.3%) Escherichia coli (EC), among which 17 were extended spectrum β lactamase, and two carbapenem-resistant Enterobacteriaceae. Among the 61 KP-PLA patients, 42(68.9%) cases were diagnosed with diabetes, 16(26.2%) cases with biliary diseases, and one (1.6%) case with malignant tumor. Among the 15 EC-PLA patients, six cases were diagnosed with diabetes, nine cases with biliary diseases, and four cases with malignant tumors. There were statistically significant differences ( χ2=4.307, 4.784 and 8.536, respectively, all P<0.05). After admission, the patients were treated with antibiotics alone or combined with drainage. One-hundred and sixty-seven (97.1%) cases got improved. Conclusions:The clinical manifestations of PLA are atypical, and the dominant pathogens are KP and EC. The risk factors of PLA are diabetes mellitus, biliary diseases and malignant tumors.
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OBJECTIVE@#To investigate the effect of interleukin-17A (IL-17A) on chemosensitivity of ovarian cancer cells to cisplatin (DDP) and explore the mechanism in light of autophagy regulation.@*METHODS@#Ovarian cancer SKOV3 cells cultured @*RESULTS@#DDP increased the expression of IL-17RA in ovarian cancer SKOV3 cells. Treatment with IL-17A significantly reduced the susceptibility of SKOV3 cells to cisplatin-induced apoptosis (@*CONCLUSIONS@#IL-17A/IL-17RA can decrease chemosensitivity of SKOV3 cells to DDP by upregulating DDP-induced autophagy.
Subject(s)
Female , Humans , Antineoplastic Agents/therapeutic use , Apoptosis/drug effects , Autophagy/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Cisplatin/pharmacology , Drug Resistance, Neoplasm/drug effects , Interleukin-17/pharmacology , Ovarian Neoplasms/drug therapy , Receptors, Interleukin-17ABSTRACT
Objective To study the regulatory role of microRNA-16 (miR-16) on human pulmonary surfactant associated protein (SP).Method Human alveolar epithelial A549 cells were transfected by miR-16 analogue,analogue negative control,inhibitor and inhibitor negative control.Blank control group was also set up at the same time.The proliferative abilities of the cells in each group were measured using cell counting kit-8 (CCK8) test.The expressions of miR-16,SP-A,SP-B and SP-C mRNA were examined using reverse transcription polymerase chain reaction (RT-PCR).The protein levels of SP-A,SP-B and SP-C were examined using western blotting method.Result RT-PCR showed that the expression of miR-16 after transfected with miR-16 analogue (34.11± 1.79) was higher than the negative control group (1.65 ± 1.07) and the blank control group (1.07 ±0.50).The expression of miR-16 after transfected with miR-16 inhibitor (0.36±0.05) was lower than the negative control group (0.96±0.13) and the blank control group (1.05±0.20).The differences were significant (all P＜0.05),and indicated that miR-16 over-expression and suppression were successfully achieved.Compared with the blank control group,cell proliferation at different time points in the analogue negative control group and the inhibitor negative control group showed no significant differences (all P＞0.05).Compared with the blank control group (1.02±0.19,1.01±0.09,1.01± 0.12) and the analogue negative control group (1.08±0.24,1.00±0.14,1.00±0.05),miR-16 down-regulated the mRNA expressions of SP-A,SP-B and SP-C (0.58±0.16,0.67±0.05,0.61±0.12).On the other hand,compared with the blank control group (1.02±0.19,1.01±0.09,1.01±0.12) and the inhibitor negative control group (1.05±0.22,0.99±0.13,0.98±0.10),miR-16 up-regulated the mRNA expressions of SP-A,SP-B and SP-C (1.66±0.33,1.29±0.11,1.23±0.12)(all P＜0.05).The trends of protein level of SP-A,SP-B and SP-C were related to their mRNA expression.Conclusion This study indicates that miR-16 inhibits pulmonary surfactant associated protein in A549 cells.
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Objective To investigate the differences in expression profile of long non-coding RNA (lncRNA) in retina between normal newborn mice and those with oxygen-induced retinopathy (OIR) to lay a foundation for further study of regulatory mechanisms of lncRNA in retinal neovascularization, and to provide a new theoretical basis for the prevention and treatment of retinopathy of prematurity (ROP). Methods A total of 48 cleaning grade C57BL/6J neonatal mice were randomly divided into two groups with 24 in each: OIR group and control group. Those in the OIR group were exposed to hyperoxia [(75±2)％ O2] from 7th to 12th day after birth, and then re-exposed to normoxia (room air) for five days to establish a OIR mouse model. Mice in the control group were raised in room air all the time. Retinal tissues were collected from each mouse on the 17th day after birth. Retinal angiography and hematoxylin-eosin (HE) staining were performed to confirm the establishment of OIR mouse model. Expression profiles of lncRNAs were analyzed by lncRNA high-throughput sequencing and the results were verified by real-time fluorescence quantitative polymerase chain reaction (PCR). Based on the expression of lncRNAs and mRNA and the relationship between their genomic locations, potential target genes of lncRNAs were obtained. Gene ontology analysis (GO) and Kyoto Encyclopedia of Genes and Gnomes (KEGG) pathway analysis were used for bioinformatics analysis of lncRNA expression profile in OIR. T test and Graphpad prism were used for statistical analysis and spreadsheet. Results Retinal angiography and HE staining identified serious retinal damage in the OIR group. In total, 1118 differentially expressed lncRNAs ( ≥ 2.0-fold difference in expression, P<0.05) relating to OIR between the two groups were identified by lncRNA high-throughput sequencing. Results of real-time fluorescence quantitative PCR confirmed that XLOC_150632 and XLOC_150636 were upregulated in the OIR group as compared with those in the control, while XLOC_122045,XLOC_100454, XLOC_170009 and XLOC_122042 were downregulated. GO analysis showed that 852, 1148 and 2100 target genes of differentially expressed lncRNAs were relating to biological process, cellular component and molecular function, respectively. KEGG analysis showed that the target genes were associated with 36 biological pathways, mainly relating to retinal development, chemotaxis, migration and energy metabolism of vascular endothelial cells. Conclusions lncRNAs are differentially expressed in healthy newborn mice and newborn mice with OIR. Fibroblast growth factor 2 (FGF2), the potential target gene of XLOC_150632 and XLOC_150636, is related to phosphatidylinositol-3-kinase (PI3K) signaling pathway and may be involved in retinal neovascularization. This might be a potential target for ROP prevention and treatment.
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Newcastle disease virus is paramyxoviridae, Avian mumps virus genus type I, and infects more than 250 species of birds, causing huge losses on poultry farming worldwide. Numerous experiments have demonstrated that Newcastle disease virus has oncolytic activity on tumor cells and can selectively replicate in cancer cells. Thus, Newcastle disease virus is a potential therapeutic agent for cancer treatment. Some human clinical trials achieved good results. In this review, we summarized research progress of the relationship between the structural protein of Newcastle disease virus and virulence, anti-tumor and autophagy of Newcastle disease.
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Objective@#To compare the dosimetric parameters of target volumes and organs at risk between volumetric-modulated arc therapy (VMAT) and intensity-modulated radiotherapy (IMRT) for left sided breast cancer after breast-conserving surgery by a meta-analysis.@*Methods@#Literature search was performed to include related studies to analyze the dosimetric parameters of target volumes and organs at risk.@*Results@#A total of 11 studies involving 154 patients were included in meta-analysis. There were no significant differences in Dmean, HI of PTV-Whole breast and PTV-Boost. Comparing to IMRT, VMAT increased the conformity index (CI) of PTV-Whole breast (P=0.000) and PTV-Boost (P=0.002). When the mean volume of target volumes was≤634 cm3, there were no significant differences in Dmean, V5, V20 of the heart and left sided lung, V30 of the heart, Dmean of the right sided lung and breast between VMAT and IMRT.When the mean volume of target volumes was>634 cm3, the Dmean(P=0.037), V5(P=0.005) and V20(P=0.030) of the heart in IMRT was lower than VMAT, but the V30(P=0.001) of heart in VMAT was lower than IMRT.IMRT showed significantly lower Dmean(P=0.013), V5(P=0.000), V20(P=0.000) of the left sided lung, and Dmean(P=0.001) of the right sided lung than VMAT.There were no significant differences in Dmean of the right sided breast.@*Conclusions@#There were no significant differences in dosimetric parameters of target volumes between VMAT and IMRT.When the mean volume of target volumes was≤634 cm3, there were no significant differences in dosimetric parameters of organs at risk. When the mean volume of target volumes was>634 cm3, IMRT has some advantages in protection of the heart and lung.
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Objective Although intersphinctoric resection is the standard method for clinical treatment of local low rectal cancer, it is lack of control for the occurrence and development of postoperative incontinence symptoms. Therefore, the effects of different ISR on postoperative stool function were analyzed retrospectively to provide guidance for judging the degree of fecal incontinence. Methods A total of 73 cases of ISR patients with low rectal adenocarcinoma treated in the oncology ward, and the outpatient department of our department from January 2014 to November 2017 were retrospectively studied. According to the distance of the tumor from the anal margin and different ISR procedures, 73 cases were divided into three group:group ISR (37 cases), subtotal ISR group (28 cases) and complete ISR group (8 cases). The anus dynamic index, Wexner subjective score of incontinence and multiple regression analysis were used to compare the severity of fecal incontinence in each group before and after operation to determine the function of stool control. Results There was no significant difference in the maximum resting pressure of anal canal, HPZ, the maximum systolic pressure of the anal canal and the Wexner in the three groups. Multiple variable ROC model regression analysis was used to analyze the influencing degree on the function of the stool, and the results showed that the Wexner score, HPZ, the maximum resting pressure of the anal canal, and the maximum systolic pressure had an obvious effect on the ability to control the stool, in which the HPZ correlated most predominately (P<0.01, HR=1.74, 95％CI 1.49- 2.03), and the age and sex factors were not selected in the equation. Conclusions Inter sphincter resection combined with new adjuvant therapy can effectively improve the anal fecal control ability of patients with low rectal cancer. The change of HPZ is of significance for prejudging the severity of fecal incontinence, which is worthy of further study.
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Objective To analyze the effect of custom abutment and computer aided design and computer aided manufacturing(CAD/CAM) provisional crown on gingival shaping around the implant,and to research the main effect of the custom abutment in the formation of gingival.Methods According to the wishes of patients,fifteen patients were divided into two groups.Gingival molding used CAD/CAM in the observation group(n=7) to design and make custom abutment and provisional crown.The control group(n=8) used the prefabricated abutment and the hand-made self-curing resin temporary crown.Immediately after inserting of final insertion and three months later,pink esthetic score(PES) was calculated in both two groups.Results The labial gingiva curve of the observation group was (1.93±0.27)points,which was higher than (1.56±0.51)points of the control group,there was significant difference between the two groups(t=2.496,P0.05).The PES score of the observation group was (8.50±1.09)points,which was higher than (7.75±1.13)points of the control group,but the difference was not statistically significant(P>0.05).Conclusion The effect of the custom abutment on curvature of facial mucosa is significant.The combination of custom abutment and digital provisional crown can achieve better soft tissue stability and ideal aesthetic effect in a short time.
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Objective To explore the role of miR-20a on pulmonary surfactant synthesis of alveolar epithelial cells A549 and its potential mechanism.Methods Lentivirus miR-20a overexpression vector(miR-20a group) or lentivirus no-load vector(no-load group) was transfected into A549 cells,and the expression of green fluorescent protein(GFP) was observed to determinate the transfection effficiency;cell proliferation was detected by using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT);the bioinformatics software and database were applied to predict and analyze the target genes of miR-20a about lung development;expressions of miR-20a,pulmonary surfactant-associated protein A(SP-A),pulmonary surfactant-associated protein B(SP-B),pulmonary surfactant-associated protein C(SP-C) and pulmonary surfactant-associated protein D(SP-D) mRNA were detected by using quantitative real-time PCR(qPCR);the expressions of SP-A protein,SP-B protein,SP-C protein,SP-D protein and protein signal transducers and activators of transcription 3 (STAT3) were detected by using Western blot.Results Observation of GFP expression under a fluorescent microscope indicated similar transfection efficiency,and real time-PCR showed that the expression of miR-20a increased after being transfected with lentivirus miR-20a overexpression vector(3.85 ± 0.18)compared with the normal group (0.99 ± 0.04)and the no-load group (1.21 ± 0.12),and the differences were significant(t =10.85,9.64,all P ＜0.001).As a result,lentivirus miR-20a overexpression vector was constructed successfully.Online software predicted that STAT3 gene was likely to be the target gene of miR-20a.Compared with the normal group (24 h,48 h,72 h:0.23 ± 0.01,0.39 ± 0.01,0.56 ± 0.03) and the no-load group (24 h,48 h,72 h:0.25 ± 0.01,0.44 ± 0.05,0.59 ± 0.01),miR-20a did not change the cell proliferation at different time points(24 h,48 h,72 h:0.26 ± 0.01,0.41 ± 0.02,0.58 ± 0.02) (all P ＞ 0.05).Compared with the normal group (1.00 ± 0.05,1.24 ± 0.20,1.31 ± 0.09,0.89 ± 0.12) and the no-load group (0.76 ± 0.10,1.31 ± 0.13,1.50 ± 0.11,1.01 ± 0.11),miR-20a up-regulated the mRNA expressions of SP-A,SP-B,SP-C and SP-D (2.05 ± 0.17,2.14 ± 0.10,2.84 ± 0.09,1.66 ± 0.08),and the differences were significant (all P ＜ 0.05).Compared with the normal group (0.46 ± 0.01,0.27 ± 0.03,0.69 ± 0.01,0.43 ± 0.01) and no-load group (0.43 ± 0.01,0.21 ± 0.01,0.79 ± 0.02,0.44 ± 0.02),miR-20a also increased the protein expressions of SP-A,SP-B,SP-C and SP-D (0.55 ±0.01,0.47 ±0.05,0.96 ±0.02,0.59 ±0.03),the diffe-rences were statistically significant (all P ＜0.05).The expression of STAT3 in miR-20a group(0.37 ±0.05) was significantly lower than that in the normal group(0.60 ±0.04) and the no-load group (0.68 ±0.06),and the differences were statstically significant (all P ＜ 0.05) in A549.Conclusions STAT3 is a downstream target gene of miR-20a.miR-20a can promote pulmonary surfactant synthesis of alveolar epithelial cells A549 by inhibiting STAT3.
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Background Previous studies showed that the pathogenesis of uveitis is related to γδ T cells.However,it remains unclear that how these cells are involved in experimental autoimmune uveitis (EAU).Objective This study aimed to observe the dynamic changes of γδ T cells in EAU and explore the role of γδ T cells in the pathological process of EAU.Methods Forty-five C57BL/6(B6) mice were assigned to the normal control group (six mice) and EAU model group (thirty-nine mice).The mice were immunized subcutaneously at 6 spots on the footpads,tail base,and flank with emulsion containing human interphotoreceptor retinoid binding protein1-20 (IRBP1-20) emulsified in complete Freund's adjuvant.After immunization,the mice were examined for clinical signs of EAU by using a Genesis-D camera.The changes of histopathology were compared by hematoxylin and eosin staining.Mouse lymphocytes were isolated and purified from the spleens of IRBP1-20-immunized or normal B6 mice by using a γδ T-cell isolation kit.Flow cytometry was used to detect the changes of intracellular expression of interleukin-17A (IL-17A),and then transferred the activated γδ T cells into EAU models to analyze the changes of clinical signs and histopathology of EAU.Experimental study program as well as the use and feeding of the animals were authorized by the Animal Management and Use Committee of Shandong Traditional Chinese Medicine University.Results The inflammatory symptoms in mouse eyes appeared on day 12 after modeling.The initial changes were fundal blood vessel thickening and minimal inflammatory cell infiltration.Then,multifocal chorioretinal lesions,serious vasculitis and linear lesions were observed on days 16-20,along with abundant lymphocyte infiltration in the vitreous and retinal disorganization.The inflammation symptom scores and the pathological inflammation scores at different time points after modeling had statistically significant differences (F =51.399,P =0.000;F =47.342,P =0.000).The inflammatory symptoms in the eyes began to abate from day 28 onwards.The number of γδ T cells was obviously increased during the inflammation phase of EAU at day 16-20 after modeling,with the number of γδ T cells was (5.67 ±-0.49) ％ and (5.78 ±±0.55) ％,respectively,which was significantly higher than (1.53 ± 0.14) ％ before modeling,with significant differences between them (both at P＜0.05),meanwhile CD69 levels and the integrin lymphocyte function-associated antigen-1 (LFA-1) and secreted IL-17A were elavated.The secretion level of IL-17A was (13.40±0.50)％ and (17.80±2.37)％ on day 16 and day 20 after modeling,respectively,which was significantly higher than (1.53 ± 0.19) ％ before modeling,with significant differences between them (P =0.000,0.001).The activated γδ T cells were transferred into EAU model,the inflammation symptom scores were 1.00 (1.00,2.00) after activated γδ T cells were transferred into EAU model,which was significantly higher than 0.75 (0.05,1.00) of the untransferred group (Z =27.00,P =0.03),and the symptoms of EAU were aggravated.Conclusions The proportion of γδ T cells reaches peak in inflammation of EAU,and the cells are activated.The activated γδ T cells in the EAU model play a immune regulation role by secreting IL-17A.
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Objective To study the roles of miR-20a in lipopolysaccharide induced inflammation of A549 cells and the possible mechanisms.Method The miR-20a mimic/inhibitor were transfected into A549 cells, and the cells were stimulated using lipopolysaccharide for 24 h.Interleukin-6 ( IL-6) and IL-8 were detected at mRNA level and protein level using real-time PCR and ELISA method , respectively.Protein expression of apoptosis signal regulating kinase 1 (ASK1)、P38、P-P38、JNK and P-JNK were detected using Western blot. Result Compared to mimic negative control group , the levels of mRNA and protein expression of IL-6 and IL-8 in the mimic group were all significantly decreased ( P<0.05).Compared to inhibitor negative control group , the levels of mRNA and protein expression of IL-6 and IL-8 in the inhibitor group were all significantly increased (P<0.05).The levels of ASK1, P-P38 and P-JNK protein in the mimic group were significantly lower than the mimic negative control group (P<0.05);the level of protein expression of ASK1, P-P38 and P-JNK in the inhibitor group were all higher than the inhibitor negative control group (P<0.05).Conclusion The regulation of ASK1 by miR-20a may play an important role in the inflammation process of acute respiratory distress syndrome .
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Objective To explore the clinical effect of amiodarone combine with irbesartan on arrhythmia patients with heart failure ,to observe its effect on cardiac function and its mechanism .Methods 124 patients with chronic heart failure combine with arrhythmia were selected in the hospital from September 2013 to November 2014 ,they were randomly divided into the control group and the observation group ,62 patients in each group .The control group was treated with amiodarone on the basis of routine thera-py ,the observation group was treated with amiodarone combine with irbesartan .The clinical efficacy of the two groups were as-sessed after treated 6 months ,the maintenance of sinus rhythm were observed after 1 ,3 and 6 months .By ultrasound heartbeat to assess heart function :cardiac stroke volume (SV) ,left ventricular ejection fraction (LVEF) ,left ventricular end systolic diameter (LVESD) and left ventricular end diastolic diameter (LVEDD) ,and to observe the adverse reactions .Results The efficiency rate of the observation group was 88 .71% (55/62) ,higher than that of the control group 62 .90% (39/62) ,the difference was statistically significant (P 0 .05) .Conclusion Amiodarone combine with irbesartan has good clinical efficacy in the treatment of heart failure combine with arrhythmia ,the maintenance rate of sinus rhythm can be significantly enhanced ,and improve heart function ,it has a higher se-curity .