As an important part of tumor microenvironment, neutrophils are poorly understood due to their spatiotemporal heterogeneity in tumorigenesis. Here we defined, at single-cell resolution, CD44-CXCR2- neutrophils as tumor-specific neutrophils (tsNeus) in both mouse and human gastric cancer (GC). We uncovered a Hippo regulon in neutrophils with unique YAP signature genes (e.g., ICAM1, CD14, EGR1) distinct from those identified in epithelial and/or cancer cells. Importantly, knockout of YAP/TAZ in neutrophils impaired their differentiation into CD54+ tsNeus and reduced their antitumor activity, leading to accelerated GC progression. Moreover, the relative amounts of CD54+ tsNeus were found to be negatively associated with GC progression and positively associated with patient survival. Interestingly, GC patients receiving neoadjuvant chemotherapy had increased numbers of CD54+ tsNeus. Furthermore, pharmacologically enhancing YAP activity selectively activated neutrophils to suppress refractory GC, with no significant inflammation-related side effects. Thus, our work characterized tumor-specific neutrophils in GC and revealed an essential role of YAP/TAZ-CD54 axis in tsNeus, opening a new possibility to develop neutrophil-based antitumor therapeutics.
Humans ; Animals ; Mice ; Adaptor Proteins, Signal Transducing/metabolism* ; Transcription Factors/metabolism* ; Stomach Neoplasms/pathology* ; Neutrophils/pathology* ; Signal Transduction/genetics* ; YAP-Signaling Proteins ; Tumor Microenvironment ; Hyaluronan Receptors/genetics*

Objective:To explore the current status and the influencing factors of knowledge, attitude and behavior of radiodermatitis in patients with nasopharyngeal carcinoma undergoing radiotherapy, so as to provide a scientific basis for nursing staff to formulate effective health education programs.Methods:A self-designed questionnaire was used to investigate 220 radiotherapy patients with nasopharyngeal carcinoma in the radiotherapy department of 4 tertiary A hospitals in Hunan Province.Results:The scores of knowledge, attitude, and behavior of radiodermatitis patients with nasopharyngeal carcinoma were (61.58±19.93), (75.70 ±15.64), (65.87±14.21) points, respectively. The main factors influencing of behavior are knowledge, attitude, radiodermatitis grade, radiotherapy frequency, and family personal monthly income level ( t values were 1.978-8.081, P<0.05). Conclusion:At present, patients with nasopharyngeal carcinoma undergoing radiotherapy have a partial understanding of radiodermatitis and poor self-observation of radiodermatitis. Nursing staff should pay special attention to the patients with incomplete knowledge, negative attitudes, low family personal monthly income, low frequency of radiotherapy, and low grade of radiodermatitis.

Objective:To evaluate changes in the estimate glomerular filtration rate(eGFR)with aging and the risk factors.Methods:A retrospective cross-sectional study was performed based on people receiving physical examinations at the First Affiliated Hospital of Nanjing Medical University from January 2017 to January 2018.Subjects were divided into seven subgroups according to age: 18-29, 30-39, 40-49, 50-59, 60-69, 70-79, and ≥80 years old.eGFR was estimated by the Chronic Kidney Disease Epidemiology Collaboration(CKD-EPIScr)equation.Multivariate linear regression was used to analyze the correlation between eGFR and the influencing variables.The chi-square test was used to compare the incidences of eGFR<60 ml·min -1·1.73m -2in different age groups. Results:A total of 33 824 participants were included in this study.There was a negative linear eGFR-age correlation in the subjects.The mean annual rate of decline in eGFR was 0.83 ml·min -1·1.73m -2.Furthermore, the decline was steady and accelerated from the third and seventh decade onward( F=9.51, 5.37, both P=0.000). Multiple linear regression analysis showed that aging was the most prominent factor( β=-0.604, -0.534, both P=0.000), followed by serum uric acid(BUA)( β=-0.270, -0.280, both P=0.000), fasting blood-glucose(FBG)( β=-0.064, -0.046, both P=0.000), systolic blood pressure(SBP)( β=-0.015, -0.028, both P<0.05), and diastolic blood pressure(DBP)( β=-0.010, -0.026, both P<0.05). In non-elderly subjects, eGFR was found to have negative associations with body mass index(BMI)and albumin(ALB)( β=-0.028, -0.047, all P=0.000). However, in the elderly, eGFR was positively associated with ALB( β=0.022, P=0.031). eGFR showed no statistically significant correlation with BMI, TC and LDL-C.The prevalence of eGFR<60 ml·min -1·1.73m -2increased with age, at 1.55%(523/33 824)for all subjects, of whom 73.80%(386/523)were aged over 60.The incidence obviously increased from 0.22%(14/6 453)for aged 18-29 to 22.57%(214/948)for aged 80 and above( χ2=2433.71, P=0.000). Conclusions:eGFR decreases significantly with age.The incidence of eGFR<60 ml·min -1·1.73m -2in the elderly is high; eGFR is significantly correlated with BUA, FBG, SBP, DBP, and ALB in the elderly.

OBJECTIVE To detect potential mutations of GCDH gene in five patients with glutaric acidemia type I (GA-I). METHODS Genomic DNA was extracted from peripheral blood samples from the patients. The 11 exons and their flanking sequences of the GCDH gene were amplified with PCR and subjected to direct sequencing. RESULTS Four mutations of the GCDH gene were identified among the patients, which included c.532G>A (p.G178R), c.533G>A (p.G178E), c.106_107delAC (p.Q37fs*5) and c.1244-2A>C. Among these, c.1244-2A>C was the most common, while c.106_107delAC was a novel mutation, which was predicted to be pathogenic by MutationTaster software. CONCLUSION The diagnosis of GA-I has been confirmed in all of the five patients. Identification of the novel GCDH mutations has enriched the mutational spectrum of the GCDH gene.

OBJECTIVETo investigate the mutations of SLC22A5 gene in patients with systemic primary carnitine deficiency (CDSP).

METHODSHigh liquid chromatography tandem mass spectrometry (HPLC/MS/MS) was applied to screen congenital genetic metabolic disease and eight patients with CDSP were diagnosed among 77 511 samples. The SLC22A5 gene mutation was detected using massarray technology and sanger sequencing. Using SIFT and PolyPhen-2 to predict the function of protein for novel variations.

RESULTSTotal detection rate of gene mutation is 100% in the eight patients with CDSP. Seven patients had compound heterozygous mutations and one patient had homozygous mutations. Six different mutations were identified, including one nonsense mutation [c.760C>T(p.R254X)] and five missense mutations[c.51C>G(p.F17L), c.250T>A(p.Y84N), c.1195C>T(p.R399W), c.1196G>A(p.R399Q), c.1400C>G(p.S467C)]. The c.250T>A(p.Y84N) was a novel variation, the novel variation was predicted to have affected protein structure and function. The c.760C>T (p.R254X)was the most frequently seen mutation, which was followed by the c.1400C>G(p.S467C).

CONCLUSIONThis study confirmed the diagnosis of eight patients with CDSP on the gene level. Six mutations were found in the SLC22A5 gene, including one novel mutation which expanded the mutational spectrum of the SLC22A5 gene.

Adult ; Amino Acid Sequence ; Base Sequence ; Cardiomyopathies ; diagnosis ; genetics ; metabolism ; Carnitine ; deficiency ; genetics ; metabolism ; DNA Mutational Analysis ; methods ; Female ; Gene Frequency ; Genotype ; Humans ; Hyperammonemia ; diagnosis ; genetics ; metabolism ; Infant, Newborn ; Male ; Muscular Diseases ; diagnosis ; genetics ; metabolism ; Mutation ; Organic Cation Transport Proteins ; genetics ; metabolism ; Reproducibility of Results ; Sensitivity and Specificity ; Sequence Homology, Amino Acid ; Solute Carrier Family 22 Member 5 ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo detect potential mutations in six patients with citrullinemia.

METHODSGenomic DNA was extracted from peripheral blood samples from the patients. Mutations of the ASS1, ASL and SLC25A13 genes were screened using microarray genotyping combined with direct sequencing.

RESULTSOne patient was diagnosed with argininosuccinate lyase deficiency, and has carried a homozygous c.1311T>G (p.Y437*) mutation of the ASL gene. The remaining five patients were diagnosed with neonatal intrahepatic cholestasis due to citrin deficiency, and have respectively carried mutations of the SLC25A13 gene including [c.851-854delGTAT+c.851-854delGTAT], [c.851-854delGTAT+IVS6+5G>A], [c.851-854delGTAT+IVS16ins3kb], [c.851-854delGTAT+IVS6-11A>G] and [c.851-854delGTAT+c.1638-1660dup23]. Among these, the c.1311T>G mutation was first identified in the Chinese population, and the IVS6-11A>G mutation was a novel variation which may affect the splicing, as predicted by Human Splicing Finder software.

CONCLUSIONThis study has confirmed the molecular diagnosis of citrullinemia in six patients and expanded the mutational spectrum underlying citrullinemia.

Argininosuccinate Lyase ; genetics ; Argininosuccinate Synthase ; genetics ; Citrullinemia ; genetics ; DNA Mutational Analysis ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Mitochondrial Membrane Transport Proteins ; genetics ; Mutation

Objective To explore the clinical features of respiratory syncytial virus(RSV) pneumonia in full-term neonatal patients.Methods All 422 full-term newborns diagnosed as pneumonia in NICU of Shenzhen Children's Hospital during January 2014 to January 2015 were included in this study.They had been detected for RSV in the way of direct immunofluorescence assay.According to the detection results, they were divided into RSV positive group and RSV negative group, the clinical data in two groups were analyzed.Results Forty-five cases were RSV positive,377 cases were RSV negative.The proportion of breast feeding was 42.22％ vs.65.25％ ,the proportion of cesarean section was 20.00％ vs.76.12％ in two groups,there were significant differences between the two groups.Hospitalization time, birth weight, gestational age, the age of admission showed no difference between two groups.The incidencs of cough (100％), shormess of breath (88.89％), three depressions (48.89 ％), fine rales (66.67 ％), wheezing (22.22％) in RSV positive group were higher than those in the RSV negative group(84.88％ ,42.44％, 13.26％, 13.53％ ,3.98％ respectively), there were significant differences between the two groups.The incidences of fever, saliva, nasal showed no significant difference between the two groups.There was significant difference in the X-ray chest film performance between two groups,RSV positive group was more emhrysema(71.11％ vs.6.9％) ,and less patch shadow(88.89％ vs.93.10％).The laboratory examination of blood routine test, C-reactive protein,respiratory failure, the positive rate of sputum culture, pneumothorax, pleural effusion were without differences.Conclusion RSV is an important pathogen of full-term neonates with infectious pneumonia.Breastfeeding and eutocia can reduce the incidence of RSV infection.Cough, shortness of breath, pulmonary rales, and emphysema in X-ray were common in RSV pneumonia.

Objective To investigate the role of signal transduction of Toll-like receptors (TLRs) in inflammatory response of neonatal sepsis.Methods Twenty children with neonatal sepsis and 16 health neonates were studied.Real-time PCR were used to evaluate the levels of TLR1 ～ TLR10,myeloid differentiation protein 2 (MD-2),myeloid differentiation factor 88 (MyD88),interleukin (IL)-1β,IL-6,IL-8 and tumor necrosis factor-α (TNF-α) mRNA expression in peripheral blood mononuclear cells.Expressions of proinflammatory cytokines (IL-1β,IL-6,IL-8,TNF-α) were measured by ELISA.Results (1) Compared with control group,the mRNA levels of TLR2,TLR4 in neonatal sepsis group were up-regulated significantly (TLR2:55.16±12.78 vs 9.53 ± 3.73,P ＜ 0.01 ;TLR4:125.22 ±30.64 vs 23.17 ± 5.78,P ＜0.01),the differences were not significant as to other TLRs.(2) Transcription levels of MD-2 and MyD88 were significantly up-regulated in neonatal sepsis group (MD-2:376.83 ± 62.16 vs 12.92 ± 2.54,P ＜ 0.01 ; MyD88:11.97 ±2.48 vs 2.77 ±0.59,P ＜0.01).(3) Expressions of proinflammatory cytokines (IL-1β,IL-6,IL-8,TNF-α) in neonatal sepsis group were higher than those of control group [PCR:(IL-1β:21.72 ± 5.56 vs 5.69 ± 1.26,P ＜0.01 ;IL-6:71.39 ± 18.34 vs 9.65 ±2.13,P ＜0.01 ;IL-8:29.39 ±6.72 vs 8.72 ± 1.95,P＜0.01;TNF-α:65.42 ± 16.95 vs 12.33 ±3.45,P ＜0.01).ELISA:(IL-1β:2 977.36 ±653.97 vs 480.52 ± 120.36,P ＜ 0.01 ; IL-6:3 143.82 ± 775.08 vs 393.78 ± 96.55,P ＜ 0.01 ; IL-8:2 510.78 ± 686.77 vs 276.91 ±72.46,P ＜0.01 ;TNF-α:3 582.24 ± 876.13 vs 1 233.68 ± 289.39,P ＜ 0.01)].Conclusion Abnormal activation of TLRs and higher expressions of proinflammatory cytokines in neonatal sepsis,suggesting that aberrant activation of TLRs may be one of the initiating factors of immune aberrance in neonatal sepsis.

Objective To assess the relationship between BRAF gene mutations and clinical phenotype of malignant melanoma.Methods Tissue specimens were collected from the lesions of 80 patients with malignant melanoma,and from the normal skin of 30 patients with trauma in the Department of Plastic Surgery or General Surgery,and subjected to paraffin embedding and DNA extraction.PCR was performed to amplify the exon 11 and 15 of BRAF gene followed by DNA sequencing.Chi-square test and Fisher's exact test were carried out to assess the relationship between BRAF gene mutations and clinical phenotypes of malignant melanoma.Results BRAF gene mutations were found in 19 (23.8％) of the 80 malignant melanoma specimens.Among the 19 mutationpositive specimens,17 (88.2％) carried mutations in exon 15 of BRAF gene with V600E as the most frequent (88.2％,15/17) mutation type,and 2 (10.5％) carried mutations in exon 11.No mutation was found in any of the normal skin tissue specimens.The average age at onset was 57.5 years in these patients.The frequency of BRAF gene mutation was significantly higher in patients younger than 60 years than in those older than 60 years (37.1％ vs.13.3％,x2=6.613,P ＜ 0.05).A significant difference was observed in the frequency of BRAF gene mutation among tissue specimens of mueosal,acral and non-aeral malignant melanoma (18.2％ (4/21) vs.14.7％(5/34) vs.41.7％ (10/24),x2=6.167,P ＜ 0.05).There was no significant association between BRAF gene mutation and gender,race or lymph node metastasis (all P ＞ 0.05).Conclusions BRAF gene is a hot spot for mutations in patients with malignant melanoma in Xinjiang Uygur Autonomous Region,with V600E point mutation in exon 15 as the most frequent mutation type.BRAF gene mutations appear to be closely correlated with the age at onset of and lesional sites in,but uncorrelated with gender and race of or lymph node metastasis in,patients with malignant melanoma.

Objective To dynamically measure cardiac output in shock neonates and to explore its clinical values.Methods The cardiac output and stroke volume of left ventricle in 30 shock neonates and 30 normal neonates were measured by noninvasive ultrasonic cardiac output monitor (USCOM) and the measurement was repeated in shock neonates after volume expansion and dopamine(2-4μg/(kg·min))treatment.Results The cardiac output and stroke volume of left ventricle in shock neonates ((0.36 ± 0.24) L/min,(3.5 ± 1.5) cm~3) were significantly lower than those in normal neonates ((0.49 ± 0.15) L/min,(2.7 ± 1.6) cm~3)) and however were improved significantly after volume expansion and dopamine treatment ((0.52 ± 0.28) L/min,(3.8 ± 1.8) cm~3).The urine volume was also increased significantly after treatment (P < 0.05).Conclusions Dynamic measurement of the cardiac output in early shock neonates might offer guidance on clinical fluid resuscitation.