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 establish and evaluate a fluorescent antibody to membrane antigen (FAMA) method for detecting antibodies against varicella-zoster virus (VZV) based on Vero E6 cells. 【Methods】 Based on the adapted VZV-Oka-E6 strain that VZV-Oka live attenuated varicella vaccine strain grew in Vero E6 cells, Vero E6 cells were infected with VZV-Oka-E6 of three different doses (10^4.65, 10^4.95 and 10^5.25 TCID50), and the cytopathic effect was observed under a microscope to determine the optimal infection dose of VZV-Oka-E6 strain. Then the detectable sensitivity of the infected cell antigen slides prepared after fixing the infected cells with different fixatives was compared to determine the optimal fixative. As a result, the FAMA method based on Vero E6 cells for the determination of neutralizing anti-VZV has been developed. The established FAMA assay was used to detect the international standard for varicella zoster immunoglobulin with different titers to determine the sensitivity of the assay. The specificity of the assay was evaluated by detecting specific antibodies against human herpes simplex virus (HSV) type 1 and type 2. The neutralizing anti-VZV antibodies of the international standard for varicella zoster immunoglobulin were detected using VZV-infected cell antigen slides prepared in the same batch and four different batches, respectively, to determine the intra-assay repeatability and inter-assay repeatability. The international standard for varicella zoster immunoglobulin with three known titers was detected to evaluate the relative accuracy of this assay. The anti-VZV titers in 20 apheresis plasma samples were determined with the newly established FAMA test and ELISA test, respectively, and the detection results of the two methods were compared using Spearman’s correlation test. 【Results】 The optimal infection dose of the VZV-Oka-E6 strain in FAMA assay was determined to be 10^5.25 TCID50, and acetone precooled at -20℃ was used as the fixative. The FAMA test has a high sensitivity with a detecting limit of 31.25 mIU/mL for neutralizing anti-VZV titers. The negative result was observed when detecting herpes simplex virus (HSV) type 1 and 2 specific antibodies. The international standard for varicella zoster immunoglobulin was detected by VZV infected cell antigen slides prepared in the same batch and 4 different batches, with the coefficient of variation being 29.95% and 26.71%, respectively. The detection value of the international standard for varicella zoster immunoglobulin with three different titer levels was consistent with their theoretical value. The correlation coefficient of the detection results of 20 apheresis plasma samples by the FAMA test and ELISA test was 0.268. 【Conclusion】 The VZV FAMA assay has good sensitivity, specificity, repeatability, and relative accuracy in detecting neutralizing anti-VZV titers. It can be applied for detecting neutralizing anti-VZV titers in apheresis plasma samples as well as the varicella-zoster immunoglobulin (VZIG) preparations.

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 investigate the protective effect of dihydromyricetin (DHM) on doxorubicin (DOX)-induced myocardial injury and its mechanism.Methods:Twenty-four healthy male SD rats were divided into 4 groups: control group, DOX group, DOX+DHM100 group and DOX+DHM200 group. Echocardiography was used to measure cardiac function. At the end of the 6th week, the rats were anesthetized and sacrificed, and the pathological changes of the cardiac tissues were observed by HE staining, Masson staining and WGA staining. Cardiomyocyte apoptosis was observed by TUNEL staining, and protein levels of NLRP3, caspase-1, IL-1β, bax and bcl-2 were detected by Western blot and immunohistochemistry.Results:Compared with the control group, the left ventricular ejection fraction and left ventricular fractional shortening decreased significantly in DOX group, while left ventricular internal dimension at systole and left ventricular internal dimension at diastole increased. In DOX+DHM group, both left ventricular ejection fraction and left ventricular fractional shortening increased, while left ventricular internal dimension at systole and left ventricular internal dimension at diastole decreased ( P<0.05). Furthermore, DOX group showed significant myocardial injury histologically, while DOX+DHM group significantly inhibited DOX-induced myocardial injury in rats. Meanwhile, cardiomyocyte hypertrophy was found in the DOX group, while the cardiomyocyte hypertrophy was notably inhibited in the DOX+DHM group. Compared with the control group, the apoptotic rates of cardiomyocytes and the levels of bax/bcl-2　ratio were significantly increased in DOX group, which were significantly alleviated in the DOX+DHM group ( P<0.05). In addition, the levels of NLRP3, caspase-1 and IL-1β were increased as compared with control group, while the levels of the above indicators were remarkably reversed in DOX+DHM group as compared with DOX group ( P<0.05). Conclusion:DHM alleviates DOX-induced myocardial injury in rats by inhibiting NLRP3 inflammasome and reducing cardiomyocyte apoptosis.

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.

Objective To explore the situation and risk factors of peripherally inserted central catheter (PICC)-related infections among neonate so as to provide a nursing reference for preventing catheter-related infections. Methods From September 2015 to June 2017, a prospective study was carried out to 811 neonates with PICC from 7 hospitals in Shenzhen City to observe the incidence of catheter-related infections. Simple correlation and multiple factors unconditional Logistic regression was used to analyze the correlative factors of catheter-related infections. Results Among 811 neonates, there were 770 (94.9%) without and 41 (5.1%, 1.95/1 000 catheter days) with catheter-related infections along with 20 cases with exit-site infection and 21 cases with catheter related bloodstream infections (CRBSI). Top three pathogens of CRBSI included Staphylococcus epidermidis, fungus and klebsiella pneumonia. Simple correlation showed that there were statistical differences in gestational age, birth weight of the neonate, disinfection methods of infusion connector, sterile protective barrier during maintenance of catheter between infection group and non-infection group (χ2=4.026,4.964, 4.369,7.463;P＜ 0.05). Multiple factors unconditional Logistic regression revealed that the risk factor contained birth weight＜ 1 200 g (OR=2.099, 95%CI: 1.103-3.996, P＜ 0.05), and the protective factors consisted of sterile protective barrier during maintenance of catheter (OR=0.393, 95%CI: 0.206-0.749,P＜0.01). Conclusions Birth weight ＜1 200 g, sterile protection during maintenance of catheter are the influencing factors of neonatal PICC catheter-related infections. Sterile protective barrier during maintenance of PICC for neonate should include wearing sterile mask, round hat, using aseptic packets and wearing sterile gloving to maintain the catheter. Aseptic technique should be paid more attention to during indwelling catheter and maintaining catheter for premature with birth weight ＜1 200 g.

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

OBJECTIVETo understand the general feature of patients with Mycobacterium tuberculosis (MTB) and human immunodeficiency virus (HIV) co-infectious (TB/HIV) in Guangxi, from 2007 to 2012.

METHODSInformation regarding individuals that the contributory causes of death were due to MTB infection among HIV as the underlying cause of death from the Vital Registration System, together with bacterium smear or culture results, onset of TB, time that TB was diagnosed and entered an Internet base TB surveillance system was collected and checked. Data including information on time of death, age, occupation, the underlying cause of death among TB patients, bacterium distribution, average age of death, interval from onset to death, percentage of TB/HIV co-infection patients among all the patients etc, were all analysed.

RESULTS203 patients died from HIV associated with TB from the Guangxi Vital Registration System were identified between 2007 and 2012. The average percentage of TB/HIV co-infection cases accounted for 8.24% (ranging from 3.94% in 2007 to 13.27% in 2012) among all the deaths of HIV infection while it accounted for 9.90% (ranging from 2.56% to in 2007 to 26.88% in 2012) among patients with MTB infection in the same period. The average percentage of deaths from TB/HIV co-infection in 2010 and 2012 accounted for 10.66% (ranging from 8.83% to 13.27%)and 22.17% (ranging from 20.60% to 26.88%)among patients died of HIV and TB infection respectively. The male-female ratio was 4.21 for 1, with the average age of death as 44.65 (44.65 ± 15.52) years;median time from TB symptoms onset to diagnosis as 37 (mean 94.31, standard deviation 206.07) days, record as (94.31 ± 206.07); median time from diagnosis to death as 46 (165.22 ± 282.19) days, 54.68% TB/HIV patients died within two months of being diagnosed with TB and the median time from TB symptoms onset to death as 131 (257.68 ± 340.79) days. 16.26% of the TB/HIV cases were bacterium confirmed TB cases.

CONCLUSIONCompare to those TB patients without HIV, less bacterium evidence was found in TB/HIV patients. High burden caused by HIV disease was seen if they were co-infected with TB. An increasing proportion of deaths was noticed among patients co-infected with HIV and TB in the last three years, suggesting that the coverage of antiretroviral therapy be scaled up together with the strengthening of the capability on early TB case-finding among people live with HIV.

Adult ; China ; epidemiology ; Coinfection ; mortality ; Female ; HIV Infections ; microbiology ; mortality ; Humans ; Male ; Middle Aged ; Tuberculosis ; mortality ; virology ; Young Adult

Objective To understand the general feature of patients with Mycobacterium tuberculosis(MTB)and human immunodeficiency virus(HIV)co-infectious(TB/HIV)in Guangxi, from 2007 to 2012. Methods Information regarding individuals that the contributory causes of death were due to MTB infection among HIV as the underlying cause of death from the Vital Registration System,together with bacterium smear or culture results,onset of TB,time that TB was diagnosed and entered an Internet base TB surveillance system was collected and checked. Data including information on time of death,age,occupation,the underlying cause of death among TB patients, bacterium distribution,average age of death,interval from onset to death,percentage of TB/HIV co-infection patients among all the patients etc,were all analysed. Results 203 patients died from HIV associated with TB from the Guangxi Vital Registration System were identified between 2007 and 2012. The average percentage of TB/HIV co-infection cases accounted for 8.24%(ranging from 3.94%in 2007 to 13.27%in 2012)among all the deaths of HIV infection while it accounted for 9.90%(ranging from 2.56%to in 2007 to 26.88%in 2012)among patients with MTB infection in the same period. The average percentage of deaths from TB/HIV co-infection in 2010 and 2012 accounted for 10.66%(ranging from 8.83% to 13.27%) and 22.17%(ranging from 20.60% to 26.88%) among patients died of HIV and TB infection respectively. The male-female ratio was 4.21 for 1,with the average age of death as 44.65 (44.65 ± 15.52)years;median time from TB symptoms onset to diagnosis as 37(mean 94.31,standard deviation 206.07)days,record as(94.31 ± 206.07);median time from diagnosis to death as 46(165.22 ± 282.19)days,54.68%TB/HIV patients died within two months of being diagnosed with TB and the median time from TB symptoms onset to death as 131 (257.68 ± 340.79) days. 16.26% of the TB/HIV cases were bacterium confirmed TB cases. Conclusion Compare to those TB patients without HIV,less bacterium evidence was found in TB/HIV patients. High burden caused by HIV disease was seen if they were co-infected with TB. An increasing proportion of deaths was noticed among patients co-infected with HIV and TB in the last three years,suggesting that the coverage of antiretroviral therapy be scaled up together with the strengthening of the capability on early TB case-finding among people live with HIV.