TTYH3 is a prognostic biomarker and associates with immune cell infiltrations of lung adenocarcinoma / TTYH3 es un biomarcador pronóstico y se asocia con infiltraciones de células inmunitarias del adenocarcinoma de pulmón

SUMMARY: We investigated Tweety Family Member 3 (TTYH3) level in lung adenocarcinoma (LUAD) and its relationship with immune infiltration in tumors by bioinformatics. Differential expressions of TTYH3 in lung cancer were analyzed with Oncomine, TIMER, GEO, UALCAN and HPA. Relationship of TTYH3 mRNA/protein levels with clinical parameters was analyzed by UALCAN. Co-expressed genes of TTYH3 in LUAD were analyzed using Cbioportal. Its relationship with LUAD prognosis was analyzed by Kaplan-Meier plotter. GO and KEGG analysis were performed. Correlation between TTYH3 and tumor immune infiltration were tested by TIMER, TISIDB and GEPIA. We found that TTYH3 was significantly increased in LUAD tissues. TTYH3 high expression was closely related to poor overall survival, post progression survival and first progression in LUAD patients. TTYH3 mRNA/protein levels were significantly associated with multiple pathways. Specifically, TTYH3 up-regulation was mostly related to biological regulation, metabolic process, protein blinding, extracellular matrix organization and pathways in cancer. Moreover, TTYH3 was positively associated with immune cell infiltration in LUAD. Finally, TTYH3 was highly expressed in LUAD as revealed by meta-analysis. TTYH3 is closely related to the prognosis of LUAD and immune cell infiltration, and it can be used as a prognostic biomarker for LUAD and immune infiltration.

Investigamos por bioinformática el nivel de Tweety Family Member 3 (TTYH3) con adenocarcinoma de pulmón (LUAD) y su relación con la infiltración inmune en tumores. Las expresiones diferenciales de TTYH3 en cáncer de pulmón se analizaron con Oncomine, TIMER, GEO, UALCAN y HPA. Con UALCAN se analizó la relación de los niveles de ARNm/proteína de TTYH3 con los parámetros clínicos. Los genes coexpresados de TTYH3 en LUAD se analizaron utilizando Cbioportal. Su relación con el pronóstico LUAD se analizó mediante plotter de Kaplan- Meier. Se realizaron análisis GO y KEGG. TIMER, TISIDB y GEPIA probaron la correlación entre TTYH3 y la infiltración inmune tumoral. Encontramos que TTYH3 aumentó significativamente en los tejidos LUAD. La alta expresión de TTYH3 estuvo estrechamente relacionada con una supervivencia general deficiente, supervivencia posterior a la progresión y primera progresión en pacientes con LUAD. Los niveles de ARNm/ proteína de TTYH3 se asociaron significativamente con múltiples vías. Específicamente, la regulación positiva de TTYH3 se relacionó principalmente con la regulación biológica, el proceso metabólico, el cegamiento de proteínas, la organización de la matriz extracelular y las vías en el cáncer. Además, TTYH3 se asoció positivamente con la infiltración de células inmunitarias en LUAD. Finalmente, TTYH3 se expresó altamente en LUAD como lo reveló el metanálisis. TTYH3 está estrechamente relacionado con el pronóstico de LUAD y la infiltración de células inmunitarias, y se puede utilizar como biomarcador pronóstico para LUAD y la infiltración de células inmunitarias.

Investigation on the use of new types of drugs among HIV/AIDS patients in Ningbo City / 预防医学

Objective@#To investigate the use of new types of drugs among HIV/AIDS patients in Ningbo City, Zhejiang Province, so as to provide insights into surveillance and interventions for use of new types of drugs among HIV/AIDS patients.@*Methods@#The HIV/AIDS patients in Ningbo City were randomly sampled from the HIV/AIDS Prevention and Control Information System of Chinese Disease Prevention and Control Information System. Patients' demographics, use of new types of drugs and sexual behaviors were collected using questionnaire surveys. Patients' hair samples were collected, and new types of drugs were determined in hair using liquid chromatography-mass spectrometry (LC-MS). In addition, factors affecting the use of new types of drugs were identified using a multivariable logistic regression model.@*Results@#A total of 254 HIV/AIDS patients were enrolled, including 214 men (84.25%), 31 cases aged under 25 years (12.20%), 66 cases aged 25 to 30 years (25.98%), and 157 cases aged 31 to 75 years (61.81%). There were 30 cases reported previous use of new drugs (11.81%), including 27 cases reported previous use of methamphetamine (90.00%). There were 48 cases tested positive of new types of drugs, with a positive rate of 18.90%, including 44 cases tested positive for methamphetamine (91.67%). Multivariable logistic regression analysis identified age (25 to 30 years, OR=6.926, 95%CI: 1.412-33.969), occupation (students/teachers/cadres/employees/retirees, OR=6.971, 95%CI: 2.123-22.889; housekeeping and housework servants/unemployed, OR=3.356, 95%CI: 1.289-8.739; business/public place servants, OR=2.447, 95%CI: 1.033-4.448) and syphilis infection during recent six months (OR=3.491, 95%CI: 1.664-7.326) as factors affecting the use of new types drugs among HIV/AIDS patients. @*Conclusions @#The use of new types of drugs exists in HIV/AIDS patients in Ningbo City. Age, occupation and syphilis infection were associated with the use of new types of drugs among HIV/AIDS patients.

Effect of exposure to low concentrations of benzene on miR-223 and miR-155 expression in peripheral blood lymphocytes / 预防医学

Objective @#To investigate the effect of exposure to low concentrations of benzene on miR-155 and miR-223 expression in peripheral blood lymphocytes among workers with benzene exposure. @*Methods @#A hundred male employees at a risk of exposure to benzene (the exposed group) were randomly sampled from two small metal products manufacturing enterprises and one medium-sized chemical raw material and chemical products manufacturing enterprise in Ningbo City, Zhejiang Province, and 60 age-matched male employees without benzene exposure were randomly selected as the unexposed group. Age, body mass index ( BMI ), smoking status, alcohol consumption, disease history, medication history and routine blood testing results of subjects were collected using a questionnaire survey. The 8-hour time weighted average concentration ( CTWA ) of benzene was measured in the workplace using thermal desorption gas chromatography, and the urine 8-hydroxy-2' deoxyguanosine ( 8-OHdG ) levels were determined using high-performance liquid-chromatography tandem mass spectrometry (HPLC-MS/MS). The miR-155 and miR-223 expression was quantified in peripheral blood lymphocytes using quantitative fluorescent reverse transcription-polymerase chain reaction assay, and the factors affecting miR-155 and miR-223 expression were identified using multivariable logistic regression analysis. @*Results @#The subjects in the exposed group had a mean age of ( 31.17±7.30 ) years, and were exposed to low concentrations of benzene ( CTWA, 0.05 to 0.30 mg/m3 ) , while the subjects in the unexposed group had a mean age of ( 32.52±6.15 ) years. There were no significant differences between the exposed and unexposed groups in terms of age, BMI, proportion of smokers or proportion of alcohol consumers ( P>0.05 ). There was no significant difference in the median relative miR-155 expression between the exposed and unexposed groups ( 0.953 vs. 1.293, P>0.05 ), and lower median relative miR-223 expression was quantified in the exposed group than in the unexposed group ( 0.540 vs. 1.433, P<0.05 ). Multivariable logistic regression analysis revealed that down-regulation of miR-223 expression correlated with exposure to benzene ( OR=2.719, 95%CI: 1.308-5.651 ). @*Conclusion @#Down-regulation of miR-223 expression may be associated with exposure to low concentrations of benzene.

Evaluation of excess mortality risk related to heat wave in Ningbofrom 2013 to 2018 / 预防医学

Objective @#To evaluate the excess mortality risk related to heat wave in Ningbo, Zhejiang from 2013 to 2018, so as to provide a basis for formulating coping strategies for heat wave.@*Methods @#The data of daily mortality, meteorological and air quality from May to October in Ningbo from 2013 to 2018 were obtained from Ningbo Center for Disease Control and Prevention, Ningbo Meteorological Bureau and Environmental Monitoring Center of Ningbo, respectively. The generalized linear model ( GLM ) and distributed lag non-linear model ( DLNM ) were used to estimate the associations between heat wave and cause-specific mortality. @*Results @#Among 1 104 days of the study period, 18 heat waves occured and lasted for 132 days, accounting for 11.96%. A total of 102 954 deaths were reported in the same period. The risks of mortality in circulatory system diseases ( RR=1.09, 95%CI: 1.03-1.16 ), respiratory system diseases ( RR=1.14, 95%CI: 1.04-1.25 ), digestive system diseases ( RR=1.38, 95%CI: 1.15-1.65 ), nervous system diseases ( RR=1.32, 95%CI: 1.08-1.61 ), mental disorders ( RR=1.51, 95%CI: 1.12-2.03 ) and accidental injury ( RR=1.18, 95%CI: 1.06-1.32 ) and all causes ( RR=1.10, 95%CI: 1.06-1.14 ) increased at lag 0-1 day of heat wave. The total excess death related to heat wave was 1 218 ( 95%CI: 731-1 705 ) . The excess deaths of circulatory system diseases, respiratory system diseases, accidental injury, digestive system diseases, nervous system diseases, mental disorders, urinary system diseases and endocrine system diseases were 313 ( 95%CI: 104-556 ), 206 ( 95%CI: 59-368 ), 164 ( 95%CI: 55-292 ), 122 ( 95%CI: 48-208 ), 69 ( 95%CI: 17-131 ), 56 ( 95%CI: 13-113 ), 18 ( 95%CI: -15-64 ) and 3 ( 95%CI: -51-72 ). The excess deaths of urinary system and endocrine system diseases was not statistically significant ( P>0.05 ). @*Conclusion @#Heat wave can increase the mortality risk on the day and after a day in Ningbo from 2013 to 2018. Circulatory system diseases, respiratory system diseases and accidental injury rank top three in excess deaths.

miR-200c regulates malignant biological behaviors of triple negative breast cancer MDA-MB-231 cells via targeting cell energy metabolism and multiple signaling pathways / 中国肿瘤生物治疗杂志

@#Objective: To investigate the effects of miR-200c on the proliferation, apoptosis and migration of triple negative breast cancer cell (TNBC) MDA-MB-231 and its metabolism-related molecular mechanism. Methods: miR-200c-231 (MDA-MB-231 overexpressing miR-200c) cells, miR-NC-231 (MDA-MB-231 transfected with miRNA-negative control) and the corresponding transplanted tumor models in nude mice were used as the study subjects. qPCR was used to detect the content of miR-200c and other related genes in cells and transplanted tumor tissues. The number of Ki67 positive cells in tumor tissue was analyzed by immunohistochemistry. The migration and apoptosis of cells were examined by Transwell chamber method and Flow cytometry, respectively. The expressions of proteins associated with proliferation, migration, and metabolism related signaling pathways in cells and tissues were confirmed by Western blotting. The changes of oxygen consumption rate (OCR), extracellular acidification rate (ECAR) and metabolic phenotype were detected by Seahorse energy metabolism detector. UPLC/LTQ-Orbitrap-MStechniquewasusedtoprofilethedifference of metabolites in cells. Results: The content of miR-200c in miR-200c-231cellswassignificantlyhigherthanthatinmiR-NC-231cells.Themass ofmiR-200c-231transplantedtumornotablydecreased,andthenumberofKi67positivecellsintumortissuesalsodecreasedsignificantly. The migration ability of miR-200c-231 cells decreased and the apoptosis rate increased (all P<0.01), accompanied with declined expressionsofZEB1/2,Vimentin,cyclinD1andincreasedexpressionofcleavedPARP(P<0.05orP<0.01),aswellasdecreasedphosphorylation leverofSTAT1/3andNF-κBbutincresedphosphorylationleverofCAMP(allP<0.05).OverexpressionofmiR-200cinMDA-MB-231cells increasedOCRandthecontentof10antitumor metabolites, but decreased ECAR and tryptophan 2,3-plus dioxidase (TDO2) expression (P<0.05 or P<0.01). Conclusion: miR-200c targeting TDO2 elevates the level of intracellular anticancer metabolites in TNBC MDAMB-231 cells, promotes the transformation from glycolysis to aerobic respiration phenotype, and inactivates STAT3 and NF-κB pathyway but activates cAMPpathway TNBC MDA-MB-231 cells, thus affects the malignant biological behaviors of MDA-MB-231 cells.

Prospective association of childhood BMI trajectory and polygenic genetic risk with puberty timing / 中国学校卫生

Objective@#To examine pubertal timing across body mass index (BMI) trajectory under polygenic susceptibility in boys and girls,and to provide a reference basis for children’s adolescent development deviation form early intervention strategies.@*Methods@#All the participants were recruited from 1 to 3 grade in 2016 from 2 Bengbu primary school and were followed up for 3 consecutive years. The study comprised 997 children (418 boys) with available data for height, weight, BMI, breast Tanner stages and testicular volume annually. The polygenic risk score (PRS) was computed based on 17 SNPs derived from published genome-wide association studies for early pubertal timing. Group-based trajectory model (GBTM) was used to identified BMI trajectory in children. Accelerated failure time model (AFT) was used to examine associations of different BMI trajectory and polygenic risk with pubertal development in boys and girls.@*Results@#Classes of BMI trajectory were persistently healthy weight, persistently overweight and persistently obesity. Adjusted concomitant variables, boys with persistently obesity exhibited 6.10-mo delay of testicular volume in low polygenic risk group (adjusted TR=1.05，P=0.04). Compared with the girls in persistently healthy weight group, the girls with low PRS were persistently overweight or obesity, which was associated with thelarche age 3.42 and 6.84-mo earlier, respectively (adjusted TR=0.97，0.94,P<0.01). Persistently overweight or obesity in girls with moderate PRS was associated with an earlier age of thelarche timing of 6.72 and 8.96-mo, respectively (adjusted TR=0.94，0.92, P<0.01). At high PRS groups, the persistently obese girls were found to have a more advanced age (10.80 and 12.96-mo, respectively) of thelarche (adjusted TR=0.90，0.88, P<0.01).@*Conclusion@#Persistently overweight and obesity is associated with early thelarche in girls, but persistently obesity may increase delayed puberty risk in boys with low polygenic risk.

Prospective association between adverse childhood experiences and pubertal development / 中国学校卫生

Objective@#To explore the impact of adverse childhood experiences on pubertal development,and to provide a reference for conducting the targeted intervention in the early stage.@*Methods@#A cluster sampling method was used to randomly select students of grades 3 to 4 from three primary schools in Ma’anshan municipality of Anhui province in October 2017. Self-reported childhood adverse experiences (ACEs), including physical, emotional and sexual abuse, as well as physical and emotional neglect were collected. Testicular volume and breast Tanner stage were assessed. A follow-up survey was conducted one year later to assess ACEs and pubertal development. ACEs of each dimension were divided into non-exposure group, transient exposure group and sustained exposure group according to ACEs exposure at baseline and follow-up. Multivariate linear regression model was used to analyze the relationship between ACEs exposure and pubertal development after 1-year follow-up.@*Results@#Sustained emotional abuse was positively associated with increased breast Tanner stage and testicular volume (breast: β=0.36, 95%CI=0.09-0.63, P<0.01; testicular volume: β=1.07, 95%CI=0.47-1.66, P<0.01); sustained physical abuse was positively associated with decreased testicular volume (β=-0.83, 95%CI=-1.58--0.08, P<0.05); sustained and transient sexual abuse were significant positively associated with decreased breast Tanner stages (sustained: β=-1.43, 95%CI=-2.86--0.02, P<0.05; transient: β=-0.45, 95%CI=-0.73--0.19, P<0.05); sustained emotional neglect was positively associated with increased breast Tanner staging (β=0.33, 95%CI=0.06-0.61, P<0.05) and decreased testicular volume (β=-0.19, 95%CI=-1.38--0.19, P<0.01).@*Conclusion@#Sustained emotional abuse was associated with early pubertal development in boys and girls,and the specitic mechanisam differences of different ACES puberty development effects and the gender differences of the same ACES puberty development effects need to be furthur explored.

Parental warmth buffers against the effect of childhood bullying victimization on internalizing symptoms / 中国学校卫生

Objective@#To investigate the longitudinal association between peer bullying victimization and internalizing symptoms, and to examine whether parental warmth buffers against this effect,so as to provide a reference for improving the mental health of the special group of children.@*Methods@#Students in grade 4 and grade 5 from three primary schools in Ma’anshan, Anhui Province were selected in Oct. 2017 with informed consent. Height, weight and pubertal development were objectively assessed. Internalizing symptoms and peer bullying victimization were evaluated at baseline and 1-year follow-up by using the MacArthur Health and Behavior Questionnaire-Child. Children were classified into three groups as non-bullying, non-consistent and consistent bullying victimization group. Multiple linear regression model was used to analyze whether the association between peer bullying victimization and internalizing symptoms varied across different parental warmth groups.@*Results@#Compared with non-bullying victimization group, consistent-and non-consistent bullying victimization had higher internalizing symptoms at baseline and 1-year follow-up(P<0.01). Students reported consistent, non-consistent bullying victimization under moderate parental warmth had lower internalizing symptoms at baseline and 1-year follow-up than those reported under low parental warmth group(P<0.01). Multiple liner regression showed that consistent bullying victimization [β=0.66(95%CI=0.33-0.99), 0.37(95%CI=0.15-0.59), 0.58(95%CI=0.31-0.84), P<0.05] and non-consistent bullying victimization[β=0.33(95%CI=0.18-0.47), 0.28(95%CI=0.13-0.41), 0.29(95%CI=0.08-0.51),P<0.05] were associated with higher 1-year follow-up internalizing symptoms in non-consistent, moderate-and low parental warmth group, while such association was not observed in consistent parental warmth group.@*Abstract@#The experience of peer bullying victimization increases the risk of internalizing symptoms and this finding suggests that parental warmth help to buffer children from the internalizing symptoms associated with bullying victimization.

Effect of Human Cytomegalovirus on Invasive Capability of Early Pregnant Extravillous Cytotrophoblasts / 华中科技大学学报（医学）（英德文版）

The effect of human cytomegalovirus (HCMV) on invasive capability of early pregnant extravillous cytotrophoblasts (EVTs) was investigated in vitro.Primary EVTs were obtained by complex phosphoesterasum digestion and gradient centrifugation from villous tissue aseptically taken from healthy pregnant women.Cytokeratin7 (CK7),vimentin (Vim) and c-erbB-2 were immunocytochemically detected to identify source of cells,and HCMVpp65 antigen was assayed to determine the infection state of primary EVTs by immunocytochemical staining.The EVTs were divided into two groups:control group and HCMV group,and the expression of c-erbB-2,matrix metalloproteinase-2 (MMP-2) and MMP-9 proteins was detected in two groups by immunocytochemistry and Western blotting.Enzymic activity changes of MMP-2 and MMP-9 were tested by gelatin zymography in primary EVTs infected with HCMV.The invasion of primary EVTs was detected by cell invasion assay in vitro after they were infected by HCMV.The cell source identification showed that the cells obtained were highly-pure primary EVTs,and primary EVTs could be infected by HCMV.Primary EVTs could express c-erbB-2,MMP-2 and MMP-9 proteins,and as compared with control group,the protein expression was decreased significantly in HCMV groups (P＜0.05).Primary EVTs could secrete active MMP-2 and MMP-9 in vitro,and the activity of two MMPs was decreased significantly in HCMV groups (P＜0.05).The in vitro cell invasion assay showed that the number of primary EVTs permeating Matrigel in HCMV group was decreased (P＜0.05).We are led to conclude that HCMV can infect primary EVTs and inhibit their invasion capability,suggesting that the impaired EVT's invasion capability might be related to the abnormal expression of c-erbB-2,MMP-2and MMP-9 proteins.

In vitro Study on Human Trophoblast Cells Infected with HCMV / 华中科技大学学报（医学）（英德文版）

Human trophoblast cells were isolated and cultured in vitro in order to investigate possible pathogenesis of intrauterine infection caused by HCMV.Trophoblast cells were obtained by compound enzymes digestion and discontinuous percoll gradient.Cells and purity were identified by using immunocytochemistry assay with anti-CK7,Vim and β-hCG antibodies.HCMV AD169 strain replication in isolated trophoblast cells and cell apoptosis were detected at different time points post infection(p.i.).The results showed that highly purified trophoblast cells were obtained.Specific virus replication was increased dramatically at the 24th h p.i.,and then increased slowly during 48 h and 72 h.Apoptosis rate of trophoblast cells infected with HCMV was(34.68±3.14)% at 24th h p.i.,while that in control group was(15.32±2.34)%(P<0.05).It was suggested that highly purified trophoblast cells can be isolated by the simplified cell purification method.HCMV can infect human trophoblast cells,and be quickly replicated,resulting in the accelerated apoptosis of human trophoblast cells during early time.