Objective:To investigate the effects of triptolide on radiosensitization of lung cancer A549 cells and the underlying mechanism.Methods:During June-September 2019, lung cancer A549 cells were treated with different concentrations of triptolide for 24 and 48 hours in Animal Experiment Center, Zhejiang Chinese Medical University, China. The inhibitory effects of triptolide on the proliferation of lung cancer A549 cells were determined using MTT method. Appropriate concentrations of triptolide and double distilled water were added to the experimental and control groups, respectively. The effects of triptolide on radiosensitization of lung cancer A549 cells was determined by colony formation assay. Radiosensitization ratio was calculated. Lung cancer A549 cells were divided into blank control, triptolide, radiotherapy, and radiotherapy + triptolide groups. The effects of triptolide on apoptosis and cell cycle of lung cancer A549 cells were determined by flow cytometry.Results:The 10% inhibitory concentration (IC 10) and half maximal inhibitory concentration (IC 50) of triptolide for treating lung cancer A549 cells were 36.61 nmol/L and 259.38 nmol/L, respectively at 24 hours, and they were 9.05 nmol/L and 61.49 nmol/L, respectively at 48 hours. Triptolide had an radiosensitization effect on lung cancer A549 cells, with the radiosensitization ratio of 1.135. The apoptosis rate in the radiotherapy + triptolide group was significantly higher than that in the radiotherapy [(45.47 ± 8.29)% vs. (5.25 ± 0.59)%, t = 6.847, P = 0.002]. The proportion of lung cancer A549 cells at the G2/M phase in the radiotherapy group was significantly higher than that in the radiotherapy + triptolide group [(27.82 ± 0.96)% vs. (11.98 ± 0.55)%, t = 20.176, P < 0.05]. The proportion of lung cancer A549 cells at the G2/M phase in the black group was significantly higher than that in the triptolide group [(17.31 ± 3.42)% vs. (8.05 ± 0.71)%, t = 3.749, P = 0.02]. Conclusion:Triptolide has a radiosensitization effect on lung cancer A549 cells, and the underlying mechanism may be related to its participation in cell apoptosis and cycle regulation.

Clustered regular interspaced short palindromic repeats (CRISPR) system has been widely used in recent years. Compared with traditional genome editing technology, CRISPR/Cas system has notable advantages, including high editing efficiency, high specificity, low cost and the convenience for manipulation. Type Ⅱ and Ⅴ CRISPR/Cas system only requires a single Cas9 protein or a single Cpf1 protein as effector nucleases for cutting double-stranded DNA, developed as genome editing tools. At present, CRISPR/Cas9 technology has been successfully applied to the genome editing of eukaryotes such as zebrafish, mice and human cells, whereas limited progress has been made in the genome editing of bacteria. In our review, we describe CRISPR/Cas system, its mechanism and summarize the optimization and progress of genome editing in bacteria.
Animals ; Bacteria ; CRISPR-Cas Systems ; Endonucleases ; Gene Editing ; Humans ; Mice

Trillions of microbes reside in the human body and participate in multiple physiological and pathophysiological processes that affect host health throughout the life cycle. The microbiome is hallmarked by distinctive compositional and functional features across different life periods. Accumulating evidence has shown that microbes residing in the human body may play fundamental roles in infant development and the maturation of the immune system. Gut microbes are thought to be essential for the facilitation of infantile and childhood development and immunity by assisting in breaking down food substances to liberate nutrients, protecting against pathogens, stimulating or modulating the immune system, and exerting control over the hypothalamic-pituitary-adrenal axis. This review aims to summarize the current understanding of the colonization and development of the gut microbiota in early life, highlighting the recent findings regarding the role of intestinal microbes in pediatric diseases. Furthermore, we also discuss the microbiota-mediated therapeutics that can reconfigure bacterial communities to treat dysbiosis.
Child ; Child, Preschool ; Disease ; etiology ; Dysbiosis ; therapy ; Gastrointestinal Microbiome ; drug effects ; Humans ; Infant ; Infant, Newborn

Objective: To investigate the effects of neonatal intensive care unit (NICU)-centered regional neonatal transport network (NTN) on the treatment of retinopathy of prematurity (ROP). Methods: A retrospective analysis was conducted to analyze the transfer, treatment and outcomes of 406 preterm infants with ROP who were transferred to the Bayi Children's Hospital Affiliated to the Seventh Medical Center of PLA General Hospital via the NTN from July 2008 to December 2014. Independent sample t-test, Chi-square test and Mann-Whitney nonparametric test were used for statistical analysis. Results: Among the 406 premature infants who were transferred to our hospital because of ROP, there were 257 males and 149 females with the gestational age of (29.5±1.9) weeks (24⁺⁵-36 weeks) and the birth weight of (1 234.8± 268.9) g (580-2 400 g). The age at transfer was (48.9±18.5) d (15-78 d) and the transport distance was (216.5±78.6) km (10-625 km). No death was reported during the transportation. Very preterm births and very low birth weight infants (VLBWI) accounted for 88.7% (360 cases) and 82.5% (335 cases), respectively. Lesions occurred in 1, 2 and 3 zones were detected in 98 (24.1%), 286 (70.4%) and 22 (5.4%) cases, respectively. Lesions of stage 1-5 were observed in 51 (12.6%), 156 (38.4%), 183 (45.1%), 12 (3.0%) and 4 (1.0%) cases. There were 186 cases (45.8%) with Plus lesions. A total of 252 cases (62.1%) underwent surgery which were achieving treatment standards, including 165 (65.5%) undergoing laser therapy, 93 (36.9%) receiving anti-vascular endothelial growth factor (VEGF) therapy, 16 (6.4%) having vitrectomy and two (0.8%) having scleral buckling. Eighteen infants were diagnosed with retinal structural abnormalities, among which 16 had retinal detachment before admission (12 in phase 4 and four in phase 5). Two without retinal detachment on admission developed to phase 4 after surgery. The incidence of retinal structural dysplasia was 0.9% (2/236) in infants receiving laser and/or anti-VEGF therapy in our hospital. Compared with the cured patients (n=234), those with retinal structural dysplasia (n=18) had a late transfer time [80 (38-270) vs 50 (19-150) d, Z=3.387, P<0.001]. Conclusions The NICU-centered and regional transfer-based ROP treatment network provides timely and effective treatment for infants with ROP.

Objective To summarize the prenatal diagnosis and genetic counseling of Turner syndrome fetuses with 46,X,i(X)(q10).Methods Two gravidas admitted to the Obstetrics and Gynecology Hospital of Dalian were enrolled in this study.One gravida,who was admitted in October 2016,was classified as high risk of Down syndrome based on prenatal serologic screening and systematic ultrasonography,which found remarkably shorter humeri and femora than fetus of the same gestations.The other was suggested to be monosomy X after non-invasive prenatal testing and admitted in November 2017.Fluorescence in situ hybridization (FISH) and karyotyping were performed for prenatal diagnosis.Peripheral blood karyotyping was also offered to the two women and their partners.Results FISH test for amniotic fluid did not find numerical abnormality in 13,18,21,and sex chromosomes in these two fetuses.Karyotype analysis showed that the two fetuses were both 46,X,i(X) (q10),while their parents were normal.Both cases were terminated after genetic counseling.Conclusions Prenatal serological screening,systematic ultrasonography and non-invasive prenatal testing may help to identify Turner syndrome fetus of 46,X,i(X) (q10).Timely and accurate prenatal diagnosis may prevent the affected fetus from being born.

Objective To investigate the clinical characteristics and prognosis of cognitive impair ment with transient ischemic attack (TIA)/minor stroke in elderly patients.Methods 115 elderly patients with TIA/minor stroke were enrolled prospectively from January 2015 March 2017,It is divided into two groups,including Non cognitive impairment group (55 cases) and non dementia vascular cognitive impairment (VCIND) group (60 cases),following for 1,3 and 6 months,com pared the clinical characteristics and the score of Montreal cognitive assessment (MoCA) scale.Results The plasma homocysteine level in VCIND group was significantly higher than non cognitive impairment group [(23.38 ± 3.31)μmol/l vs (17.21± 2.10) μmol/l,P＜0.01].The total MoCA score in non cognitive impairment group was 27.46 ± 1.21,VCIND group was 22.21±2.09,the differences between the two groups were statistically significant (P＜0.05).The average score of visual space/executive ability,delayed memory,language and abstract ability in VCIND group was lower than non cognitive impairment group (P＜0.05,P＜0.01).1,3 and 6months follow-up of group VCIND,The rate of cognitive improvement was 8.3％,33.3％ and 61.7％ (P＜0.05).Conclusion cognitive impairment in elderly patients with TIA and minor stroke is mainly manifested in the visual space/executive ability,delayed recall,language fluency/repetition and abstraction,the cognitive impairment of patients will improve over time.

Objective To identify the epidemiological changes in invasive fungal infection (IFI) in a neonatal intensive care unit (NICU) to provide information for prevention and treatment of IFI.Methods A total of 102 cases who were diagnosed with IFI among 42 187 neonates hospitalized in the NICU of Affiliated BaYi Children's Hospital,Chinese People's Liberation Army General Hospital from January 1,2009 to December 31,2014 were enrolled in this study.Since January 1,2012,the divisions of our NICU were more specific and intravenous fluconazole was administered as a routine preventive measure for high-risk infants.Clinical information of the IFI cases including general features,incidence,distribution of pathogens and drug (Amphotericin B,Fluconazole,Flucytosin,Itraconazole and Voriconazole) sensitivity were analyzed between former period (January 1,2009 to December 31,2011) and latter period (January 1,2012 to December 31,2014) by Chi-square test.Results The total incidence of IFI was 2.42‰ (102/42 187),and among the 102 IFI cases,73.5％ (75/102) were preterm infants and 75.5％ (77/102) were low birth weight infants.The incidence ofIFI in the latter period was lower than that in the former period [1.8‰ (48/26 046) vs 3.3‰ (54/16 141),x2=9.329,P＜0.01].The incidences of IFI in neonates with gestation age ＜28,≥ 28-＜32 and ≥ 32-＜37 weeks in latter period were decreased as compared with those in former period [10.6 ‰ (3/284) vs 76.9 ‰ (9/117),x2=12.569;6.1‰ (13/2 134) vs 21.9‰ (28/1 277),x2=16.868;1.4‰ (12/8 706) vs 1.9‰ (10/5 256),x2=7.165] (all P＜0.01).Altogether 103 pathogen strains were identified from 102 IFI cases as one Candida parapsilosis strain and one Laurent cryptococcus strain were both isolated from one patient.The most prevalent three pathogens were Candida albicans [51.5％ (53/103)],Candidaparapsilosis [24.3％ (25/103)] and Candida glabrata [8.7％ (9/103)].The isolated rates of Candida albicans and Candida glabrata strains in the latter period were higher than those in the former period [63.3％ (31/49) vs 40.7％ (22/54),x2=5.218;18.4％ (9/49) vs 0.0％ (0/54),x2=10.868],while the isolated rate of Candida parapsilosis strain was lower in the latter period than that in the former period [12.2％(6/49) vs 35.2％(19/54),x2=7.355] (all P＜0.05).All pathogen strains were sensitive strains except one Candida krusei strain which was isolated in the former period and was resistant to Fluconazole.Conclusions Premature infants born at lower gestational ages or with low birth weights are still at high-risk of IFI,but the incidence of IFI has declined in recent years.Routine administration of fluconazole in high-risk infants in NICU could prevente IFI without increasing drug resistance.Candida albicans is the predominant pathogen ofIFI.

Objective To explore and build a real, objective, comprehensive clinical nursing individual performance indicators architecture model,and check the rationality and validity for prize distribution by clinical application. Methods The methods included that discuss new ideas of nursing performance management by multi- disciplinary experts,developed clinical personal nursing staff performance evaluation program,worded out indicators and methods for the clinical assessment of individual nurses and nurse managers respectively,then applied research in the pilot departments and hospital step by step. Results A personal performance evaluation framework model was constructed, which include clinical nurses and nursing managers. Experimental results show that the nursing staff in this regard performance program have a high degree of recognition, 98.82% (1 741/1 762) nursing staff understanding of the purpose and significance, 97.15%(1 712/1 762) nurses think the performance model structure is reasonable. After the implementation of the performance program, the outstanding rate of personal performance appraisal of nurses was 93% (1 639/1 762). Conclusions The application of scientific performance appraisal programs can play a positive role in helping improve the quality of clinical care, and promote the stable development of the care team.

Objective To investigate the molecular regulatory mechanism of glucagon like peptide 1 (GLP-1) on proliferation, differentiation and apoptosis of human umbilical cord blood endothelial progenitor cells (EPCs). Methods EPCs were isolated from the umbilical cord blood of healthy pregnant women and cultured in 6-hole cell plate at 2×105 density in vitro, transfected with empty vector plasmid (control group), pcDNA3-GLP-1 plasmid (GLP-1 group), pcDNA3-GLP-1plasmid+AMD3100 (GLP-1+AMD3100 group) and simple AMD3100 (AMD3100 group). The pcDNA3-GLP-1 was transfected into EPCs. The 25μmol/L AMD3100 was used to block the SDF-1/CXCR4 signal pathway of EPCs for 1 h. The cell proliferation was determined by MTT method. The mRNA expressions of differentiation and apoptosis related genes PPARγ, C/EBPα and Caspase-3 were investigated by RT-PCR, and Caspase-3 activity was determined by Caspase-3 activity assay kit. Results Compared to control group, AMD3100 inhibitor showed no effects on cell proliferation, differentiation and apoptosis, while over-expression of GLP-1 in EPCs obviously promoted cell proliferation, and differentiation related genes PPARγand C/EBPαmRNA expression, but down-regulated mRNA expression and the activity of Caspase-3 significantly (P＜0.05), indicating that GLP-1 increased proliferation and differentiation of EPCs while decreased cell apoptosis. When the SDF-1/CXCR4 signaling pathway was blocked by AMD3100, over-expression of GLP-1 induced promotion of cell proliferation, and the differentiation was decreased significantly and the apoptosis was significantly increased (P＜0.05). Conclusion These data confirm that GLP-1 might promote EPCs proliferation and differentiation, and inhibit cell apoptosis through the regulation of the SDF-1/CXCR4 signaling pathway.

Objective To analyze the clinical effect of small incision operation and traditional operation in treatment of pediatric hernia,and to provide a theoretical basis for the clinical treatment of pediatric hernia.Methods 68 children with pediatric hernia were treated as the research subjects,they were randomly divided into control group and observation group,34 children in each group.The control group adopted the traditional operation treatment,while the observation group was taken small incision.The clinical efficacy and complications of both two groups were compared. Results The operation time,bleeding volume,hospitalization time and incision length of the observation group were (10.45 ±1.35)min,(5.21 ±0.78)mL,(1.98 ±0.76)d,and (1.04 ±0.23)cm,which were better than those of the control group [(15.67 ±3.24)min,(8.56 ±1.87)mL,(3.78 ±1.12)d,and (2.54 ±0.65)cm,respectively],the differences were significant (t =5.760 0,5.747 7,4.171 7,7.986 8,all P <0.05).The incidence rate of complica-tions of the observation group was 2.94%,which was significantly lower than that of the control group (11.76%),the difference was significant (χ2 =5.7118,P <0.05).Conclusion The clinical curative effect of using small incision in the treatment of pediatric hernia is significant,this method has the advantages of small operation wound,less pain, faster postoperative recovery,less complications and other advantages,it is worthy of clinical application.