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@#[摘 要] 目的：探讨miR-216b-5p对食管癌Eca109细胞顺铂（DDP）耐药性的影响及其作用机制。方法:采用qPCR法检测miR-216b-5p在食管癌细胞TE-1、KYSE-150、Eca109和耐药细胞Eca109/DDP中的表达水平。利用脂质体转染技术分别将miR-216b-5p mimic及mimic NC、自噬相关蛋白5（ATG5）过表达质粒转染到Eca109/DDP细胞中，用CCK-8、EdU法和FCM分别检测转染后细胞的增殖和凋亡；mRFP-eGFP-LC3双荧光标记实验检测mRFP-eGFP-LC3慢病毒感染后各组细胞自噬发生情况，WB法检测自噬相关蛋白LC3、Beclin 1和P62表达。用荧光素酶报告基因实验验证miR-216b-5p与ATG5的靶向关系，WB法检测ATG5的表达。建立裸鼠Eca109/DDP细胞移植瘤模型，观察miR-216b-5p过表达对移植瘤生长的影响。结果：miR-216b-5p在TE-1、KYSE-150、Eca109和Eca109/DDP细胞中均呈低表达（均P<0.05）。过表达miR-216b-5p可显著抑制Eca109/DDP细胞的增殖并诱导凋亡（均P<0.05），减少细胞中自噬小体数量（P<0.05），下调LC3Ⅱ/LC3Ⅰ比值和Beclin 1蛋白水平、上调P62蛋白水平（均P<0.05）。双荧光素酶报告基因实验证实miR-216b-5p靶向并负调控ATG5的表达（P<0.05），过表达ATG5可使miR-216b-5p mimic对Eca109/DDP细胞增殖、自噬的抑制作用和凋亡的诱导作用明显减弱（均P<0.05），自噬相关蛋白P62表达降低、LC3Ⅱ/LC3Ⅰ比值和Beclin 1表达升高（均P<0.05）。荷瘤实验结果表明，miR-216b-5p过表达可显著抑制裸鼠移植瘤的生长（P<0.05）。结论：miR-216b-5p过表达可逆转食管癌Eca109/DDP细胞对DDP的耐药性，其机制可能与靶向负调控ATG5表达并影响细胞自噬有关。
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Objective@#To understand the changes of children s height, weight, blood pressure and gender differences, to explore the relationship between overweight, obesity and childhood hypertension, and to provide a scientific basis for childhood hypertension prevention.@*Methods@#Physical examination data during 2013 to 2018 of ten primary school students in Shenzhen were collected. Growth rate of height, weight and blood pressure by age and gender were calculated. The generalized estimating equation was used to analyze the association between overweight, obesity and hypertension.@*Results@#Weight, body mass index (BMI) and systolic pressure of boys and girls increased with age ( t/Z =3.89-31.52, P <0.05). The height growth rate of girls was higher than that of boys at the age of 8-11, and reaches the peak of height growth at the age of 10, while boys were two years later than girls(boys:7.68 cm, gilrs:7.42 cm). Weight and blood pressure growth rates were similar. At the same time, the growth rate of height and blood pressure had a synchronous trend, and the peak of the growth rate of blood pressure was also at the peak stage of height growth. The OR value of obesity on childhood hypertension was 1.62(1.48-1.81), and the OR value of overweight on childhood hypertension was 2.01(1.75-2.30), both P <0.01.@*Conclusion@#There are gender differences in children s height, weight, and blood pressure, and the growth rate of height and blood pressure shows a synchronous trend. Overweight and obesity in children can increase the risk of high blood pressure and hypertension.
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Objective@#To investigate the mutual prediction relationship between peer relationship and internalizing problem.@*Methods@#One-year follow up survey was conducted with a sample of 220 preschool children from 4 kindergartens in Shandong province. The quality of peer relationship and degree of internalizing problem were evaluated with the Peer Relationship Scale and the Child Behavior Checklist Cross-lagged panel analysis was used for mutual prediction among variables.@*Results@#The quality of preschool children’s peer relationship showed an increasing trend with grade(F=6.40, 4.81, P<0.01), while the degree of internalizing problem showed a downward trend(F=7.65, 5.46, P<0.01). The predictive effect of pre-test peer relationship and internalizing problem on post-test corresponding behaviors were all statistically significant (β=0.56, 0.49, P<0.01). The predictive effect of pre-test peer relationship on post-test internalizing problem was statistically significant(β=-0.19, P<0.05).@*Conclusion@#Both peer relationship and internalizing problem has a certain stability across time, and early peer relationship and internalizing problem could predict later corresponding behaviors. Early peer relationship can predict later internalizing problem, while early internalizing problem cannot predict later peer relationship.
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Limitation in the number of insulin-producing pancreatic b-cells is a typical feature of diabetes. It has been indicated thatactivating pancreatic transcription factors can promote the transformation of hepatocytes into insulin-secreting b-like cells,indicating that direct hepatocyte differentiation seems promising as a treatment for diabetes. Nevertheless, the reprogramming efficiency still remains low. Our previous study found that the expression of c-fos-induced growth factor (FIGF)was increased in the pancreatic tissues in partial pancreatectomy mice compared to that in normal mice. Here, we observedthat treatment with Ad-FIGF was found to enhance MafA and Ngn3-induced reprogramming of BNL CL.2 cells to b-likecells with the ability of secreting insulin. And FIGF overexpression increased the levels of histone H3/H4 acetylation atMafA and Ngn3 promoter regions in BNL CL.2 cells. Importantly, in vivo study further confirmed that forced expression ofFIGF facilitated the insulin expression and decreased the blood glucose levels in STZ mice. These results strengthen thepossibility of developing cell-based therapies for diabetes through utilizing b-like cells derived from non-insulin-secretingcells.
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SUMMARY OBJECTIVE To investigate the relations of T lymphocytes, cytokines, immunoglobulin E, and nitric oxide with otitis media with effusion (OME) in children and their clinical significances. METHODS Fifty children with OME treated in our hospital were enrolled in the study (observation group). Fifty healthy children were selected as control. The percentages of CD4+ and CD8+ T lymphocyte and CD4+/CD8+ ratio in peripheral blood, and the levels of cytokine (IL)-2, IL-4, IL-6, immunoglobulin E (IgE) and nitric oxide (NO) in peripheral blood and middle ear effusion (MEE) in both groups were detected. The correlations of these indexes with OME were analyzed. RESULTS The percentage of peripheral blood CD4+ and CD8+ levels, CD4+/CD8 ratio, IgE, and NO levels in the observation group were significantly higher than those in the control group (P < 0.01). In the observation group, the IL-2 and IL-6 levels, and IgE and NO levels in the MEE were significantly higher than those in peripheral blood (P < 0.01). In addition, in the observation group, the MEE IL-2 and IL-6 levels were positively correlated with peripheral blood CD4+/CD8+ ratio, respectively r = 0.366, P = 0.009; r = 0.334, P = 0.018. CONCLUSIONS The levels of peripheral blood CD4+ and CD8+ lymphocytes and MEE IL-2, IL-6, IgE, and NO levels are increased in children with OME. These indexes have provided significant clues for the diagnosis of OME in children.
RESUMO OBJETIVO Investigar as relações entre linfócitos T, citocinas, imunoglobulina E e óxido nítrico e a otite média com efusão (OME) em crianças e sua significância clínica. MÉTODOS Cinquenta crianças com OME tratadas em nosso hospital foram incluídas no estudo (grupo de observação). Selecionamos também 50 crianças saudáveis como controle. As porcentagens de linfócitos T CD4 + e CD8 + e a razão CD4+/CD8+ no sangue periférico, além dos níveis das citocinas IL-2, IL-4, IL-6, imunoglobulina E (IgE) e óxido nítrico (NO) no sangue periférico e de efusão no ouvido médio (MEE) de ambos os grupos foram medidos. A correlação desses índices com a OME foi analisada. RESULTADOS A porcentagem dos níveis de CD4+ e CD8 +, da razão CD4+/CD8+, de IgE e NO no sangue periférico do grupo de observação foram significativamente maiores do que no grupo controle (P < 0,01). No grupo de observação, os níveis de IL-2 e IL-6, IgE e NO em MEE foram significativamente maiores do que no sangue periférico (P < 0,01). Além disso, no grupo de observação, foi encontrada uma correlação positiva entre os níveis de IL-2 e IL-6 em MEE e a razão de CD4+/CD8+no sangue periférico, respectivamente, r = 0,366, P = 0,009; r = 0,334, P = 0,018. CONCLUSÃO Os níveis de linfócitos CD4 + e CD8 + no sangue periférico e IL-2, IL-6, IgE e NO em MEE são mais altos em crianças com OME. Esses índices forneceram evidências valiosas para o diagnóstico de OME em crianças.
Subject(s)
Humans , Male , Female , Child, Preschool , Child , Adolescent , Otitis Media with Effusion/blood , Immunoglobulin E/blood , CD4-Positive T-Lymphocytes , Cytokines/blood , CD8-Positive T-Lymphocytes , Nitric Oxide/blood , Reference Values , Tympanic Membrane/metabolism , Case-Control Studies , Lymphocyte Count , Flow CytometryABSTRACT
Objective@#To understand the emotional and behavioral problems among preschool children in Ezhou city, and to analyze its association with screen time,to provide a reference for making preventive measures.@*Methods@#1 720 children were selected from 9 kindergartens in Ezhou by cluster sampling method. Questionnaires were completed by the children’s parents or primary caregivers.@*Results@#High total difficulty score accounted for 9.8%(169/1 720) of all participants,the abnormal proportion of peer communication problems is the highest. The average screen time was (1.95±1.88) hours, among them 2.6% spent more than 2 hours. The average screen time on weekends was (2.84±2.58) hours, among them 19.1% spent more than 2 hours. The results of multiple linear regression analysis showed that weekday, weekend and total screen times was associated with children’s total difficulty score, emotional problems, conduct problems, peer interaction and hyperactivity(P<0.05). Furthermore, with the duration of screen time increased, the susceptibility of emotional and behavioral problems in these four dimensions increased.@*Conclusion@#Screen time is associated with emotional and behavioral problems among preschool children. Schools and parents should pay attention to interventions that target family TV viewing practices, and parent-child interaction and communication.
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Objective To evaluate the safety and efficacy ofdexamethasone intravitreal implant 0.7 mg (DEX) for treatment of macular edema associated with retinal vein occlusion (RVO).Methods This study was a six-month,randomized,double-masked,sham-controlled,multicenter,phase 3 clinical trial with a 2-month open-label study extension.Patients with branch or central RVO received DEX (n=129) or sham procedure (n=130) in the study eye at baseline;all patients who met re-treatment criteria received DEX at month 6.Efficacy measures included Early Treatment Diabetic Retinopathy Study (ETDRS),best-corrected visual acuity (BCVA),and central retinal thickness (CRT) on optical coherence tomography.Results Time to ＞ 15-letter BCVA improvement from baseline during the first 6 months (primary endpoint) was earlier with DEX than sham (P＜ 0.001).At month 2 (peak effect),the percentage of patients with ≥ 15-letter BCVA improvement from baseline was DEX:34.9％,sham:11.5％;mean BCVA change from baseline was DEX:10.6± 10.4 letters,sham:1.7 ± 12.3 letters;and mean CRT change from baseline was DEX:-407 ± 212 μm,sham:-62 ± 224 μm (all P＜0.001).Outcomes were better with DEX than sham in both branch and central RVO.The most common treatment-emergent adverse event was in-creased intraocular pressure (IOP).Increase sin IOP generally were controlled with topical medication.Mean IOP normalized by month 4,and no patient required incisional glaucoma surgery.Conclusions DEX had a favorable safety profile and provided clinically significant benefit in a Chinese patient population with RVO.Visual and anatomic outcomes were improved with DEX relative to sham for 3-4 months after a single implant.
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Abstract Klebsiella pneumoniae is important human and animal pathogen that causes a wide spectrum of infections. In this study, isolates from cattle nasal swabs samples were identified by 16S rRNA, and to evaluate the antimicrobial susceptibility, virulence gene carrying levels, and multilocus sequence typing of K. pneumoniae isolates. 33 isolates of K. pneumoniae were isolated and identified in 213 nasal swabs samples, of which 12 were hypervirulent K. pneumoniae strains. Extended Spectrum Beta-Lactamases genes were found in 93.4% of the strains. Of which, TEM was the most prevalent (93.4%), followed by CTX-M and SHV were 57.6% and 39.4%, respectively. A main mutation pattern of quinoloneresistance-determining region, Thr83-Ieu and Asp87-Asn in gyrA and Ser87-Ile in parC, was detected in 33 K. pneumoniae isolates. All the isolates harbored at least two virulence factor genes, with ureA (97.0%) and wabG (91.0%) exhibiting high carriage rates in 33 K. pneumoniae isolates. MLST revealed 7 sequence types, of which 3 STs (2541, 2581 and 2844) were newly assigned. Using eBURST, ST2844 and ST2541 were assigned to new clonal complex 2844. Our study provides evidence and biological characteristics of K. pneumoniae isolates from cattle upper respiratory tract in Southwest China.
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Abstract Klebsiella pneumoniae is important human and animal pathogen that causes a wide spectrum of infections. In this study, isolates from cattle nasal swabs samples were identified by 16S rRNA, and to evaluate the antimicrobial susceptibility, virulence gene carrying levels, and multilocus sequence typing of K. pneumoniae isolates. 33 isolates of K. pneumoniae were isolated and identified in 213 nasal swabs samples, of which 12 were hypervirulent K. pneumoniae strains. Extended Spectrum Beta-Lactamases genes were found in 93.4% of the strains. Of which, TEM was the most prevalent (93.4%), followed by CTX-M and SHV were 57.6% and 39.4%, respectively. A main mutation pattern of quinoloneresistance-determining region, Thr83-Ieu and Asp87-Asn in gyrA and Ser87-Ile in parC, was detected in 33 K. pneumoniae isolates. All the isolates harbored at least two virulence factor genes, with ureA (97.0%) and wabG (91.0%) exhibiting high carriage rates in 33 K. pneumoniae isolates. MLST revealed 7 sequence types, of which 3 STs (2541, 2581 and 2844) were newly assigned. Using eBURST, ST2844 and ST2541 were assigned to new clonal complex 2844. Our study provides evidence and biological characteristics of K. pneumoniae isolates from cattle upper respiratory tract in Southwest China.
Subject(s)
Animals , Cattle , Bacterial Proteins/genetics , Klebsiella Infections/veterinary , Cattle Diseases/microbiology , Drug Resistance, Multiple, Bacterial , Virulence Factors/genetics , Klebsiella pneumoniae/isolation & purification , Klebsiella pneumoniae/drug effects , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , Klebsiella Infections/microbiology , China , Virulence Factors/metabolism , Multilocus Sequence Typing , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/metabolismABSTRACT
Pseudomonas aeruginosa (PA) pneumonia is a refractory,even lethal complication in immunosuppressive individuals and immune disturbances may promote the pathological process.We aimed to investigate the regulatory T (Treg) cell activity in an immunosuppressive mice model of PA pneumonia by estimating levels of main transcription factor and the main effector of Treg cells,i.e.,Forkhead box protein 3 (FOXP3) and interleukine-10 (IL-10).Seventy-two BALB/c mice were divided into four groups randomly:control (A),PA pneumonia (B),immunosuppression (C) and immunosuppression with PA pneumonia (D).Mice were sacrificed at 4,8 and 24 h after establishing experimental models.The pathological changes of lung tissue were graded,and the FOXP3 mRNA and serum IL-10 levels were detected.Histological analysis of lung tissues showed there were no significantly pathological changes in groups A and C,but significantly pathological changes were found in groups B and D,especially in group D at 8 h (P＜0.05).The expression levels of FOXP3 mRNA in groups A and C showed no significant changes at the three time points,which were significantly lower than those in groups B and D (P＜0.05).FOXP3 mRNA levels were lowest at 4 h,and there was significant difference between groups B and D (P＜0.05).The serum levels of IL-10 in groups A and C were almost normal at the three time points,but decreased significantly in groups B and D (P＜0.05).The serum levels ofIL-10 decreased to the lowest at 8 h,especially in group D (P＜0.05).The results indicate that PA pneumonia in immunosuppressive individuals worsens rapidly,which may be associated with Treg cells function disturbance.And Treg cells may be promising as adjuvant therapeutics for PA pneumonia in immunosuppressive individuals.
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A field experiment established in 1980 was conducted to evaluate the effects of open drainage ditch applied for water removal on bacterial and fungal communities of cold waterlogged paddy soils in 2011. In this experiment, traditional plate counting and temperature gradient gel electrophoresis were employed to characterize the abundance and diversity of soil bacterial and fungal communities. Four different distances from the open drainage ditch, 5, 15, 25 and 75 m with different degrees of drainage were designed for this study. Maximum populations of culturable aerobic bacteria and fungi were at 15-m distance while minimum populations were at 75-m distance. Significant differences (p < 0.05) in fungal populations were observed at all distances from open drainage ditch. The highest diversity of the bacterial community was found at a distance of 25 m, while that of the fungal community was observed at a distance of 5 m. Sequencing of excised TGGE bands indicated that the dominant bacteria at 75-m distance belonged to anaerobic or microaerobic bacteria. Relationships between microbial characteristics and soil physicochemical properties indicated that soil pH and available nitrogen contents were key factors controlling the abundance of culturable aerobic bacteria and fungi, while soil water capacity also affected the diversity of fungal community. These findings can provide the references for better design and advanced management of the drainage ditches in cold waterlogged paddy soils.