ObjectiveDetection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability, health, and metabolic rate.After exposure to environmental stimuli, both the internal reference gene and target gene would be degraded. As a result, it is imperative to consider the accurate capture of nascent RNA and the detection of transcriptional levels of RNA following environmental stimulation. This study aims to create a Click Chemistry method that utilizes its property to capture nascent RNA from total RNA that was stimulated by the environment. MethodsThe new RNA was labeled with 5-ethyluridine (5-EU) instead of uracil, and the azido-biotin medium ligand was connected to the magnetic sphere using a combination of “Click Chemistry” and magnetic bead screening. Then the new RNA was captured and the transcription rate of 16S rRNA was detected by fluorescence molecular beacon (M.B.) and quantitative reverse transcription PCR (qRT-PCR). ResultsThe bacterial nascent RNA captured by “Click Chemistry” screening can be used as a reverse transcription template to form cDNA. Combined with the fluorescent molecular beacon M.B.1, the synthesis rate of rRNA at 37℃ is 1.2 times higher than that at 15℃. The 16S rRNA gene and cspI gene can be detected by fluorescent quantitative PCR,it was found that the measured relative gene expression changes were significantly enhanced at 25℃ and 16℃ when analyzed with nascent RNA rather than total RNA, enabling accurate detection of RNA transcription rates. ConclusionCompared to other article reported experimental methods that utilize screening magnetic columns, the technical scheme employed in this study is more suitable for bacteria, and the operation steps are simple and easy to implement, making it an effective RNA capture method for researchers.

Objective To explore the effect of open reading frame 66(C12ORF66)located at chromosome 12 on the viability of MYCN amplified NB cell lines.Methods DDatasets GSE16476 and GSE49710 in R2 database were analyzed for expression level of C12ORF66 in MYCN amplified and MYCN non-amplified NB cells and its potential correlation with the prognosis of pediatric patients.C12ORF66 mRNA expression level in normal tissue immortalized cell lines,MYCN amplified and MYCN non-amplified cell lines were detected by RT-qRCR.Transient or stable knockdown of C12ORF66 cell lines were constructed to compare the difference in real time cellular analysis(RTCA),colony formation,Ki67 positive cells between the control group and the C12ORF66 knockdown group.Results By analyzing R2 datasets,C12ORF66 level in MYCN amplified samples was significantly higher than that in MYCN non-amplified samples,and the expression of C12ORF66 was negatively correlated with the prognosis of pediatric patients(P＜0.05).C12ORF66 highly expressed in MYCN-amplified BE(2)-C and SK-N-BE(2)cell lines than in MYCN non-amplified CHLA-255 and SH-SY5Y cell lines(P＜0.001).Transient or stable knockdown of C12ORF66 resulted in significant slow down of proliferation of MYCN amplified NB cells(P＜0.001),the colony formation ability was significantly reduced(P＜0.001),and the proportion of Ki67 positive cells was significantly decreased(P＜0.05).Conclusions C12ORF66 was highly expressed in MYCN amplified clinical NB samples and cell lines which is believed to be correlated with poor prognosis of pediatric patients.C12ORF66 knockdown signifi-cantly inhibits cell viability of NB cells.

Objective:To investigate the function and underlying mechanism of cysteine and glycine-rich protein 2(CSRP2)in neuroblastoma(NB).Methods:The correlation between the expression level of CSRP2 mRNA and the prognosis of NB children in NB clinical samples was analyzed in R2 Genomics Analysis and Visualization Platform.The small interfering RNA(siRNA)targeting CSRP2 or CSRP2 plasmid were transfected to NB cell lines SK-N-BE(2)and SH-SY5Y.Cell proliferation was observed by crystal violet staining and real-time cellular analysis.The ability of colony formation of NB cells was ob-served by colony-forming unit assay.Immunofluorescence assay was used to detect the expression of the proliferation marker Ki-67.Flow cytometry analysis for cell cycle proportion was used with cells stained by propidium iodide(PI).Annexin V/7AAD was used to stain cells and analyze the percentage of cell apoptosis.The ability of cell migration was determined by cell wound-healing assay.The level of protein and mRNA expression of CSRP2 in NB primary tumor and NB cell lines were detected by Western blot and quantitative real-time PCR(RT-qPCR).Results:By analyzing the NB clinical sample databases,it was found that the expression levels of CSRP2 in high-risk NB with 3/4 stages in international neuroblas-toma staging system(INSS)were significantly higher than that in low-risk NB with 1/2 INSS stages.The NB patients with high expression levels of CSRP2 were shown lower overall survival rate than those with low expression levels of CSRP2.We detected the protein levels of CSRP2 in the NB samples by Western blot,and found that the protein level of CSRP2 in 3/4 INSS stages was significantly higher than that in 1/2 INSS stages.Knockdown of CSRP2 inhibited cell viability and proliferation of NB cells.Overexpression of CSRP2 increased the proliferation of NB cells.Flow cytometry showed that the proportion of sub-G1,G0/G1 and S phase cells and Annexin V positive cells were increased after CSRP2 deficiency.In the cell wound-healing assay,the healing rate of NB cells was significantly attenuated after knockdown of CSRP2.Further mechanism studies showed that the proportion of the proliferation marker Ki-67 and the phospho-rylation levels of extracellular signal-regulated kinases 1/2(ERK1/2)were significantly decreased after CSRP2 knockdown.Conclusion:CSRP2 is highly expressed in high-risk NB with 3/4 INSS stages,and the expression levels of CSRP2 are negatively correlated with the overall survival of NB patients.CSRP2 significantly increased the proliferation and cell migration of NB cells and inhibited cell apoptosis via the activation of ERK1/2.All these results indicate that CSRP2 promotes the progression of NB by activating ERK1/2,and this study will provide a potential target for high-risk NB therapy.

Pilar leiomyoma or piloleiomyoma is a benign neoplasm of the smooth muscle arising from the arrector pili muscle. It manifests as brown to red firm papulonodules with sites of predilection being the face, trunk, and extensor surfaces of the extremities. Histologically, the lesions exhibit ill-defined dermal tumors with interlacing fascicles of spindle cells. Some genodermatoses are characterized by the development of visceral tumors and cutaneous leiomyomatosis such as Reed’s syndrome, and hereditary leiomyomatosis and renal cell cancer (HLRCC). A 55-year-old male presented with reddish-brown papules and nodules on the face and upper back, accompanied by sharp episodic pain on the face. He had undergone nephrectomy for renal cancer 9 years ago, and his younger brother had similar cutaneous manifestation. Histopathologic findings were consistent with pilar leiomyoma, showing bundles of smooth muscle tumors in the dermis. Based on the clinical information including clinical features, past medical history, and family history, HLRCC was highly suspected.To confirm the diagnosis, whole exome sequencing was performed using peripheral blood, which revealed a novel point mutation (c.739G>A, p.Glu247Lys) in the fumarate hydratase (FH) gene. We describe a confirmed case of HLRCC, which is a genetic disorder with a potential to cause visceral cancers, which dermatologists might overlook as a benign condition.

Objective:To analyze the incidence of stillbirth and the associated factors in pregnancy among pregnant residents in Wuhan.Methods:A previous birth cohort was retrospectively reviewed. The cohort was based on Wuhan Maternal and Child Information System, and the perinatal information of pregnant residents in Wuhan from January 1, 2011, to September 30, 2017 and information of selected cases was collected, including socio-demographic characteristics, pregnant history, and healthcare information during pregnancy and labor. Data on stillbirth, including fetal death in uterus and in labor, were selected for this study. Chi-square test was adopted for comparing the differences in pregnancy-related factors between live birth and stillbirth, and binary logistic regressions for exploring the influencing factors associated with the occurrence of stillbirth. Results:A total of 509 057 deliveries in Wuhan were included in this study, including 505 839 live births and 3 218 stillbirths (3 155 after exclusion of fetal death in labor), with an overall incidence of stillbirth of 6.32‰(3 218/509 057), and an annual incidence between 4.90‰ to 8.11‰. Statistically significant differences were found between the live birth and stillbirth group in the following items: maternal age [<25 years old: 19.28% (97 544/505 839) vs 19.36% (623/3 218); 25-30 years old: 48.45% (245 077/505 839) vs 45.15% (1 453/3 218); 30-35 years old: 26.09% (131 952/505 839) vs 26.29% (846/3 218); >35 years old: 6.18% (31 266/505 839) vs 9.20% (296/3 218)], educational background [middle school or below: 22.90% (115 833/505 839) vs 22.03% (709/3 218); high school: 36.37% (183 978/505 839) vs 38.72% (1 246/3 218); college or above: 40.73% (206 028/505 839) vs 39.25% (1 263/3 218)], occupation [brainworker or professionals: 33.51% (169 514/505 839) vs 31.54% (1 015/3 218); manual or freelance worker: 66.38% (335 763/505 839) vs 68.34% (2 199/3 218)], residential area [urban area: 70.00% (354 365/505 839) vs 76.32% (2 456/3 218); rural area: 30.00% (151 474/505 839) vs 23.68%(762/3 218)], and time of conception [spring (March to May): 24.27% (122 746/505 839) vs 24.08% (775/3 218); summer (June to August): 24.09% (121 867/505 839) vs 23.87% (768/3 218); fall (September to November): 26.69% (135 012/505 839) vs 25.08% (807/3 218); winter (December to next February): 24.95% (126 214/505 839) vs 26.97% (868/3 218)] (all P<0.05), but no significant difference was found in fetal gender ( P>0.05). Besides, gravidity [once: 49.32% (249 484/505 839) vs 47.02% (1 513/3 218); over twice: 50.68% (256 355/505 839) vs 52.98% (1 705/3 218)], parity [once: 73.60% (372 316/505 839) vs 77.07% (2 480/3 218); over twice: 26.40% (133 523/505 839) vs 22.93% (738/3 218)], history of stillbirth [0.33% (842/256 355) vs 0.65% (11/1 705)], hypertensive disorders in pregnancy [3.25% (16 464/505 839) vs 5.59% (180/3 218)], first trimester vaginal bleeding [2.02% (10 251/505 839) vs 2.61% (84/3 218)], placenta previa [0.98% (4 963/505 491) vs 2.64% (53/2 009)], and oligohydramnios [2.52% (12 764/505 839) vs 1.90% (61/3 218)] differed significantly between the two groups (all P<0.05). However, no significant difference was found between the two groups in terms of the proportion of women with gestational diabetes mellitus and previous spontaneous abortion (both P>0.05). After exclusion of fetal death in labor from the 3 218 stillbirths, the same results were achieved. Binary logistic regression analysis showed that women who were over 30 years old (30-35 years old: OR=1.42, 95% CI: 1.30-1.56; >35 years old: OR=2.59, 95% CI: 2.25-2.98), with a high school degree or below (middle school or below: OR=1.37, 95% CI: 1.21-1.55; high school: OR=1.28, 95% CI: 1.16-1.42), manual or freelance worker ( OR=1.18, 95% CI: 1.07-1.31), in the urban area ( OR=1.43, 95% CI:1.31-1.57), and gravidity ≥ 2 times ( OR=1.32, 95% CI: 1.21-1.43), primiparity ( OR=1.76, 95% CI: 1.58-1.96), gestational hypertension ( OR=2.80, 95% CI: 2.40-3.27), vaginal bleeding in the first trimester ( OR=1.35, 95% CI: 1.08-1.68), placenta previa ( OR=10.86, 95% CI: 8.84-13.35) and history of stillbirth ( OR=2.27, 95% CI: 1.30-3.98) were all risk factors of stillbirth. Conclusion:Pregnant women who were over 30 years old, less educated, manual worker or freelance or with a history of adverse pregnancy, vaginal bleeding in the first trimester, hypertension in pregnancy, and placenta previa are at higher risk of stillbirth

Objective: The real-world INFORM study analyzed sociodemographics, treatment patterns and clinical outcomes for patients with newly diagnosed advanced epithelial ovarian cancer (EOC) in Australia, South Korea (S.Korea) and Taiwan preceding incorporation of poly(ADP-ribose) polymerase inhibitors into clinical practice. Methods: Retrospective data from patients diagnosed with EOC (high-grade serous EOC for Taiwan) between January 2014 and December 2018 with ≥12 months follow-up from diagnosis were analyzed descriptively. Survival was evaluated by Kaplan-Meier with two-sided 95% confidence interval (CI). Results: Of the 987 patients (Australia, 223; S.Korea, 513; Taiwan, 251), 98% received platinum-based chemotherapy (CT). In S.Korea and Taiwan 76.0% and 78.9% respectively underwent primary cytoreductive surgery; in Australia, 56.5% had interval debulking surgery. Bevacizumab was included in primary/maintenance therapy for 22.4%, 14.6% and 6.8% of patients in Australia, S.Korea and Taiwan, respectively. Patients receiving bevacizumab were high-risk (reimbursement policy) and achieved similar real-world progression-free survival (PFS) compared with CT only. Overall, the median real-world PFS (months; 95% CI) was similar across Australia (16.0 [14.63–18.08]), S.Korea (17.7 [16.18–19.27]) and Taiwan (19.1 [17.56–22.29]). Conclusion This study reveals poor prognosis despite differences in demographics and treatment patterns for patients with EOC across Asia-Pacific suggesting the need for biomarker-driven novel therapies to improve outcomes.

Eupafolin, a constituent of the aerial parts of Phyla nodiflora, has neuroprotective property. Because reducing the synaptic release of glutamate is crucial to achieving pharmacotherapeutic effects of neuroprotectants, we investigated the effect of eupafolin on glutamate release in rat cerebrocortical synaptosomes and explored the possible mechanism. We discovered that eupafolin depressed 4-aminopyridine (4-AP)-induced glutamate release, and this phenomenon was prevented in the absence of extracellular calcium. Eupafolin inhibition of glutamate release from synaptic vesicles was confirmed through measurement of the release of the fluorescent dye FM 1-43. Eupafolin decreased 4-AP-induced [Ca2+ ] i elevation and had no effect on synaptosomal membrane potential. The inhibition of P/Q-type Ca2+ channels reduced the decrease in glutamate release that was caused by eupafolin, and docking data revealed that eupafolin interacted with P/Q-type Ca2+ channels. Additionally, the inhibition of calcium/calmodulindependent protein kinase II (CaMKII) prevented the effect of eupafolin on evoked glutamate release. Eupafolin also reduced the 4-AP-induced activation of CaMK II and the subsequent phosphorylation of synapsin I, which is the main presynaptic target of CaMKII. Therefore, eupafolin suppresses P/Q-type Ca2+ channels and thereby inhibits CaMKII/synapsin I pathways and the release of glutamate from rat cerebrocortical synaptosomes.

OBJECTIVE: Cardiogenic syncope can present as a seizure. The distinction between seizure disorder and cardiogenic syncope can only be made if one considers the diagnosis. Our main objective was to identify whether patients presenting with a chief complaint (reason for visit) as seizure or syncope received an electrocardiogram in the emergency department across all age groups.METHODS: We conducted a secondary analysis of data collected in the 2010 to 2014 National Hospital Ambulatory Medical Care Survey comparing patients presenting with a chief complaint of syncope versus seizure to determine likelihood of getting an evaluation for possible life threatening cardiovascular disease. The primary endpoint was receiving an electrocardiogram in the emergency department; secondary endpoint was receiving cardiac biomarkers.RESULTS: There was a total of 144,094 patient encounters. Of these visits, 1,553 had syncope and 1,470 had seizure (60.3% vs. 44.2% female, 19.9% vs. 29.0% non-white). After adjusting for age, sex, mode of arrival and insurance, patients with syncope were more likely to receive an electrocardiogram compared to patients with seizure (odds ratio, 10.86; 95% confidence interval [CI], 8.52 to 13.84). This was true across all age groups (0 to 18 years, 56% vs. 7.5%; 18 to 44 years, 60% vs. 27%; 45 to 64 years, 82% vs. 41%; ≥65 years, 85% vs. 68%; P < 0.01 for all). Car- diac biomarkers were also obtained more frequently in adult patients with syncope patients (18 to 44 years, 17.5% vs. 10.5%; 45 to 64 years, 33.8% vs. 21.4%; ≥65 years, 47.1% vs. 32.3%; P < 0.01 for all).CONCLUSION: Patients evaluated in the emergency department for syncope received an electrocar- diogram and cardiac biomarkers more frequently than those that had seizure.
Adult ; Biomarkers ; Cardiovascular Diseases ; Diagnosis ; Electrocardiography ; Emergencies ; Emergency Service, Hospital ; Epilepsy ; Female ; Humans ; Insurance ; Seizures ; Syncope

BACKGROUND AND OBJECTIVES: Joint National Committee guidelines attempt to vary treatment recommendations for patients based on considerations of their comorbidities. The aim of the present study is to estimate the age-standardized prevalence of common comorbidities among Korean hypertension patients. SUBJECTS AND METHODS: We analyzed the Korea National Health and Nutrition Examination Survey from 2007 to 2013. Among the 58423 participants, 30092 adults, aged ≥30 yrs who completed a health examination and interview survey, were selected. The survey procedures were used to estimate weighted prevalence and odds ratios for 8 comorbidities, including obesity, diabetes mellitus, dyslipidemia, cardiovascular disease, chronic kidney disease, and thyroid disease. RESULTS: Most chronic conditions were more prevalent in adults with hypertension than in those without hypertension. Common comorbidities were obesity (60.1%), dyslipidemia (57.6%), and impaired fasting glucose (45.1%). Hypertensive patients with two or more comorbid diseases were 42.2% and those with three or more diseases were 17.7%. The age- and sex-specific prevalence of three or more comorbid diseases among male hypertension patients was significantly higher than those patients in the 30-59 (p<0.05) age group. CONCLUSION: Comorbidity is highly prevalent in Korean patients with hypertension.
Adult ; Cardiovascular Diseases ; Chronic Disease ; Comorbidity* ; Diabetes Mellitus ; Dyslipidemias ; Epidemiology ; Fasting ; Glucose ; Humans ; Hypertension* ; Joints ; Korea* ; Male ; Nutrition Surveys* ; Obesity ; Odds Ratio ; Prevalence* ; Renal Insufficiency, Chronic ; Thyroid Diseases

BACKGROUND/AIMS: Recent outbreak of hepatitis A in Korea is clearly related to the epidemiological shift of hepatitis A virus (HAV). However, nationwide seroprevalence data have been limited. This study estimated the nationwide, age- and area-adjusted anti-HAV prevalence from 2005 to 2009. METHODS: Retrospective analysis of the results of total anti-HAV test in 25,140 cases which were requested by 1,699 medical institutions throughout the nation to Seoul Clinical Laboratory from Jan. 1 2005 to Dec. 31 2009 was performed. The estimated seroprevalence was adjusted by area and age of the standard population based on the 2005 Census data from Korea National Statistical Office. RESULTS: The area-adjusted anti-HAV prevalence in the children younger than 10 years were 33.4% in 2005 and 69.9% in 2009. The most susceptible age groups to HAV infection during the last 5 years were teenagers and the young adults in their age of twenties. The area-adjusted seroprevalence in 2009 were 11.9% in the age group of 20-29 years, 23.4% in the age group of 10-19 years, 48.4% in the age group of 30-39 years. The population in 40-49 years showed geographically different seroprevalence with the lowest rate in Seoul (80%). CONCLUSIONS: The most susceptible age group to HAV infection is 10-29 years, while the young children less than 10 years showed about 70% seropositivity. The changing seroepidemiology should be monitored continuously for the proper vaccination and patient care.
Adolescent ; Adult ; Age Factors ; Aged ; Child ; Child, Preschool ; Female ; Hepatitis A/*epidemiology ; Hepatitis A Antibodies/*blood ; Hepatitis A Virus, Human/immunology ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Retrospective Studies ; Seroepidemiologic Studies