[The effect of glycopyrrolate on intestinal spasm and hemodynamics in painless colonoscopy].

Objective: To investigate the effects of glycopyrrolate on intestinal spasm and hemodynamics in painless colonoscopy. Methods: A total of 100 patients who were scheduled to undergo painless colonoscopy were selected as the study subjects and randomly divided into two groups by a computerized number method. Ten patients in both groups dropped out because of disruption of the study protocol, and 45 patients from each group were included in the final analysis. Before anesthesia induction, patients in group glycopyrrolate (group G) were injected with 0.2 mg glycopyrrolate, while those in congtrol group (group C) were injected with an equal amount of saline. The heart rate, systolic blood pressure, and diastolic blood pressure were recorded at T0 (baseline period), T1 (after anesthesia induction), T2 (colonoscopy over sigmoid colon), T3 (colonoscopy over the liver region), T4 (after the end of examination), and T5 (at the awakening phase), and the degree of intestinal spasm was assessed intraoperatively using the Likert's four-point scale. The numerical rating scale (NRS) was used to assess preoperative and postoperative pain. The incidence of adverse events was recorded. Results: The general data at baseline were not statistically different between the two groups (P>0.05). During the procedure, patients in group G had lower intraoperative intestinal spasm scores than those in group C (P=0.028). Intraoperative hypotension and bradycardia occurrence were lower in group G than in group C (P<0.05), and intraoperative norepinephrine use was also lower than in the group C (P=0.034). Postoperative visual analog scale pain scores were lower in group G (P=0.047), but patients who used glycopyrrolate had a higher proportion of dry mouth (P=0.035). Conclusion: During painless colonoscopy, preoperative administration of glycopyrrolate significantly improved intraoperative hemodynamic fluctuations, reduced the incidence of hypotension and bradycardia, and relieved postoperative pain. However, glycopyrrolate use resulted in the risk of dry mouth.

Chemical and Cellular Formation of Reactive Oxygen Species from Secondary Organic Aerosols in Epithelial Lining Fluid.

INTRODUCTION: Oxidative stress mediated by reactive oxygen species (ROS) is a key process for adverse aerosol health effects. Secondary organic aerosols (SOA) account for a major fraction of particulate matter with aerodynamic diameter ≤2.5 µm (PM2.5). PM2.5 inhalation and deposition into the respiratory tract causes the formation of ROS by chemical reactions and phagocytosis of macrophages in the epithelial lining fluid (ELF), but their relative contributions are not well quantified and their link to oxidative stress remains uncertain. The specific aims of this project were (1) elucidating the chemical mechanism and quantifying the formation kinetics of ROS in the ELF by SOA; (2) quantifying the relative importance of ROS formation by chemical reactions and macrophages in the ELF. METHODS: SOA particles were generated using reaction chambers from oxidation of various precursors including isoprene, terpenes, and aromatic compounds with or without nitrogen oxides (NOx). We collected size-segregated PM at two highway sites in Anaheim, CA, and Long Beach, CA, and at an urban site in Irvine, CA, during two wildfire events. The collected particles were extracted into water or surrogate ELF that contained lung antioxidants. ROS generation was quantified using electron paramagnetic resonance (EPR) spectroscopy with a spin-trapping technique. PM oxidative potential (OP) was also quantified using the dithiothreitol assay. In addition, kinetic modeling was applied for analysis and interpretation of experimental data. Finally, we quantified cellular superoxide release by RAW264.7 macrophage cells upon exposure to quinones and isoprene SOA using a chemiluminescence assay as calibrated with an EPR spin-probing technique. We also applied cellular imaging techniques to study the cellular mechanism of superoxide release and oxidative damage on cell membranes. RESULTS: Superoxide radicals (·O2-) were formed from aqueous reactions of biogenic SOA generated by hydroxy radical (·OH) photooxidation of isoprene, ß-pinene, α-terpineol, and d-limonene. The temporal evolution of ·OH and ·O2- formation was elucidated by kinetic modeling with a cascade of aqueous reactions, including the decomposition of organic hydroperoxides (ROOH), ·OH oxidation of primary or secondary alcohols, and unimolecular decomposition of α-hydroxyperoxyl radicals. Relative yields of various types of ROS reflected the relative abundance of ROOH and alcohols contained in SOA, which generated under high NOx conditions, exhibited lower ROS yields. ROS formation by SOA was also affected by pH. Isoprene SOA had higher ·OH and organic radical yields at neutral than at acidic pH. At low pH ·O2- was the dominant species generated by all types of SOA. At neutral pH, α-terpineol SOA exhibited a substantial yield of carbon-centered organic radicals (R·), while no radical formation was observed by aromatic SOA.Organic radicals in the ELF were formed by mixtures of Fe2+ and SOA generated from photooxidation of isoprene, α-terpineol, and toluene. The molar yields of organic radicals by SOA were 5-10 times higher in ELF than in water. Fe2+ enhanced organic radical yields by a factor of 20-80. Ascorbate mediated redox cycling of iron ions and sustained organic peroxide decomposition, as supported by kinetic modeling reproducing time- and concentration-dependence of organic radical formation, as well as by additional experiments observing the formation of Fe2+ and ascorbate radicals in mixtures of ascorbate and Fe3+. ·OH and superoxide were found to be efficiently scavenged by antioxidants.Wildfire PM mainly generated ·OH and R· with minor contributions from superoxide and oxygen-centered organic radicals (RO·). PM OP was high in wildfire PM, exhibiting very weak correlation with radical forms of ROS. These results were in stark contrast with PM collected at highway and urban sites, which generated much higher amounts of radicals dominated by ·OH radicals that correlated well with OP. By combining field measurements of size-segregated chemical composition, a human respiratory tract model, and kinetic modeling, we quantified production rates and concentrations of different types of ROS in different regions of the ELF by considering particle-size-dependent respiratory deposition. While hydrogen peroxide (H2O2) and ·O2- production were governed by Fe and Cu ions, ·OH radicals were mainly generated by organic compounds and Fenton-like reactions of metal ions. We obtained mixed results for correlations between PM OP and ROS formation, providing rationale and limitations of the use of oxidative potential as an indicator for PM toxicity in epidemiological and toxicological studies.Quinones and isoprene SOA activated nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in macrophages, releasing massive amounts of superoxide via respiratory burst and overwhelming the superoxide formation by aqueous chemical reactions in the ELF. The threshold dose for macrophage activation was much smaller for quinones compared with isoprene SOA. The released ROS caused lipid peroxidation to increase cell membrane fluidity, inducing oxidative damage and stress. Further increases of doses led to the activation of antioxidant response elements, reducing the net cellular superoxide production. At very high doses and long exposure times, chemical production became comparably important or dominant if the escalation of oxidative stress led to cell death. CONCLUSIONS: The mechanistic understandings and quantitative information on ROS generation by SOA particles provided a basis for further elucidation of adverse aerosol health effects and oxidative stress by PM2.5. For a comprehensive assessment of PM toxicity and health effects via oxidative stress, it is important to consider both chemical reactions and cellular processes for the formation of ROS in the ELF. Chemical composition of PM strongly influences ROS formation; further investigations are required to study ROS formation from various PM sources. Such research will provide critical information to environmental agencies and policymakers for the development of air quality policy and regulation.

[The mediating role of anxiety and depression for family members of ICU patients in perceived social support and fatigue].

To analyze the mediating role of anxiety and depression in perceived social support and fatigue in ICU patients' families, and to provide a theoretical evidence for alleviating their fatigue status. A total of 223 family members of ICU patients who received treatment at the Affiliated Hospital of Jiangnan University from October 2020 to April 2021 were selected as the study subjects. The general data questionnaire, perceived social support scale (PSSS), generalized anxiety disorder scale (GAD-7), patient health questionnaire (PHQ-9) and fatigue assessment instrument (FAI) were used to conduct a survey. Among 223 family members of ICU patients, 155(69.51%) had fatigue problems. There were statistically significant differences in total fatigue scores of ICU patients' family members in terms of gender, age, education level, relationship with patients, residence, payment method and per capita monthly income (P<0.05). Anxiety, depression and fatigue were negatively correlated with perceived social support (r are -0.353, -0.276 and -0.416, respectively, all P<0.01). Depression and fatigue were positively correlated with anxiety (r are 0.808 and 0.703, respectively, all P<0.01), and fatigue was also positively correlated with depression (r= 0.665, P<0.01). Anxiety and depression had a partial mediating effect on perceived social support and fatigue, and the total indirect effect size was 52.64%. Comprehensive intervention on the level of social support, anxiety and depression is helpful to improve the fatigue status of ICU patients' family members.

[Bilateral posterior shoulder dislocation with associated reverse Hill-Sachs lesions after electrical injury].

After workers suffer electric shock, the ankylosis and contraction of muscle groups around the shoulder joint are more likely to lead to posterior dislocation and fracture, of which 80% are complicated with reverse Hill-Sachs injury of humeral head. This paper reports a case of bilateral posterior dislocation of shoulder joint combined with Hill-Sachs injury caused by electric shock in the Department of Orthopedics, Affiliated Huzhou Hospital, Zhejiang University School of Medicine in August 2020. The diagnosis of left posterior shoulder dislocation was clear, and the diagnosis of right posterior shoulder dislocation was missed. The patient successfully reconstructed the stability of the shoulder joint by actively performing shoulder arthroscopic surgery to repair the joint capsule. After 6 months of follow-up, there was no further dislocation and the function was good.

[One case of extensive high-pressure injection injury of hand caused by polyurethane material].

This article reports a patient with extensive high-pressure injection injury of the hand caused by mistaken injection of polyurethane material into the index finger, who was diagnosed and treated in the Department of Orthopedics of Huzhou Central Hospital in 2019. Both the digital artery and digital nerve were involved, and the polyurethane involved the right palm along the flexor tendon sheath of the index finger and wrist. Due to the lack of X-ray development, the scope of the first debridement was small, and the blood supply to the fingertip was poor. Finally, the patient's right index finger was amputated due to infection and necrosis. MR or B-ultrasound should be perfected before operation to clarify the extent of polyurethane involvement. The initial thorough debridement or multiple debridements are necessary to improve the prognosis. If the blood supply of the fingers is poor, the blood supply can be reconstructed by skin flap transplantation.

[Epidemiological and etiological characteristics of hand, foot and mouth disease among children aged 5 years and younger in Ningbo (2016 to 2019)].

OBJECTIVE: To describe the epidemiological and etiological characteristics of hand, foot and mouth disease (HFMD) among children aged 5 years and younger in Ningbo after the access of entero-virus-A71 vaccine (2016 to 2019). METHODS: A retrospective cohort study were performed in children aged 5 years and younger in Ningbo from 2016 to 2019. Data for incidence of HFMD was collected from the National Notifiable Disease Surveillance Reporting System and the Electronic Medical Records (EMRs) System, while the demographic information was derived from the Immunization Information System. Speci-mens were detected by real-time fluorescence quantitative PCR and the Wilson method was used to estimate the incidence rate and 95% confidence interval. RESULTS: From 2016 to 2019, a total of 1 044 800 residential children were observed in this population-based cohort. In the study, 102 471 cases of HFMD were diagnosed in 2 651 081 person-years, revealing an overall incidence density of 3 865.25/100 000 person-years. There was no significant decline in the number of the cases after the vaccine was available. The number of the patients of hand foot mouth disease during the four years was 93 421, of whom 84 875 (90.85%) had only one incident record, while 8 946 (9.15%) had 2 or more cases in this period; there were 69 771 (66.06%) patients who only needed to see a doctor once for each disease, 19.92% of the patients needed to be treated twice, and 14 801 (14.02%) patients needed to go to the hospital or clinic three times or more. The incidence of HFMD showed obvious seasonality and periodicity, which mainly concentrated in April to July each year, and the epidemic cycle was 2 years; most of the cases were 1 to 3-year old children, with more cases in male. The incidence density varied across the region, with the highest density observed in Ninghai (4 524.76/100 000 person-years), followed by Xiangshan (3 984.22/100 000 person-years). In 3 748 library-conformed cases, 2 834(75.61%) were detected positive, among which enterovirus-A71, Cox-A16 and other enteroviruses accounted for 9.03%, 31.55% and 59.42%, respectively. During the study period, the cumulative coverage of enterovirus-A71 vaccine increased year by year, with the proportion of enterovirus-A71 and severe cases both gradually decreasing. CONCLUSION: The current status of hand, foot and mouth disease in Ningbo is still serious. Children under 3-year old (especially male children aged 1 year) were the key population for prevention and control. Vaccination might lead to changes in major pathogenic virus type, of which more attention should be paid to the potential impact on disease surveillance, prevention and control.

[Epidemiological study of infantile epilepsy incidence density among infants under 36 months of age in Ningbo City from 2015 to 2019].

OBJECTIVE: To describe the distribution and trend of infantile epilepsy among infants under 36 months in Ningbo, Zhejiang Province. METHODS: Using the birth cohort design, we retrospectively collected the local born infants in Ningbo national health information platform from 2015 to 2019, and took the first visit of epilepsy in the electronic medical record of the platform as the new case. The incidence density and 95% confidence interval (CI) of epilepsy were estimated by Poisson distribution. RESULTS: From 2015 to 2019, a total of 294 900 children were born in Ningbo, with male accounting for 51.92%. The total person-years of observation were 595 300, while the median follow-up person-years was 2.31 [interquartile range (IQR): 1.90]. There were 575 new onset epilepsy patients during the whole observation period. The total number of visits was 2 599, with an average of 4.52. The total incidence density was 96.59/100 000 person-years (95%CI: 88.85-104.82). The median age of onset was 13 months (IQR: 15), 0-12 months old infants had the highest incidence density (102.18/100 000 person-years), 25-36 months old infants had the lowest incidence density (89.68/100 000 person-years), and the difference was not statistically significant (P>0.05). The incidence density of male was 97.58/100 000 person-years, female was 95.53/100 000 person-years, and the difference was not statistically significant (P>0.05). Fenghua was the highest (130.54/100 000 person-years, 95%CI: 94.47-175.83) and Ninghai was the lowest (66.44/100 000 person-years, 95%CI: 47.02-91. 19), with significant difference (P < 0.05). There was no significant difference in the incidence density in different birth years (P>0.05). There was significant difference in the incidence density between 0-12 months old infants in different calendar years (Ptrend < 0.05). In this age group, the incidence density was the lowest in 2015 (69.41/100 000 person-years, 95%CI: 41.79-108.39), and the highest in 2019 (225.61/100 000 person-years, 95%CI: 186.10-271.03). There was no significant difference in the incidence density between 13-24 and 25-36 months old infants in different calendar years (P>0.05). CONCLUSION: The incidence density of epilepsy in 0-36 months old infants in Ningbo City from 2015 to 2019 was low as a whole, and there was no difference in age group, gender, and year of birth. The incidence density of 0-12 months old infants increased with the year.

Research Progress on Pathological Fibrosis of Sinoatrial Node.

ABSTRACT: Human heart rhythm is mainly regulated and controlled by the sinoatrial node. Fibrosis plays an important regulating role in adjusting the structural and functional integrity of the sinoatrial node pacemaker complex. In physiological state, the fibrosis degree of sinoatrial node is negatively correlated with heart rate, positively correlated with age and heart size, and can maintain a relatively stable heart rate. Pathological fibrosis of sinoatrial node can induce various types of arrhythmias which can result in sudden death. Determination of the mechanisms related to sinoatrial node pathological fibrosis could provide a target for clinical treatment of sinoatrial node fibrosis and diagnosis basis for forensic pathologists. This paper reviews the main mechanism of sinoatrial node pathological fibrosis, including abnormal activation of cardiac fibroblast cells in sinoatrial node, hyperplasia of epicardial adipose tissue, calcium clock disorder, artery stenosis, etc., introduces the test methods, diagnostic criteria as well as its role in sudden cardiac death and discusses the potential application, to provide reference for relevant research and application.

A case of coronavirus HKU1 encephalitis.

Coronaviruses are widespread in nature and can infect several different species, causing mainly respiratory and enteric diseases. The respiratory involvement of human coronaviruses has been clearly established since the 1960s. Three of the six coronaviruses that infect humans have been shown to be neuroinvasive and neurotropic in humans: HCoV-229E, HCoV-OC43, and SARS-CoV. No reports exist on the detection of HCoV-HKU1 in the human central nervous system (CNS). We report a case of a patient, in whose cerebrospinal fluid (CSF) was detected Coronavirus NL63/HKU1. Coronavirus HKU1 was detected in the sputum. With effective antiviral therapy and the use of glucocorticoids, the patient was eventually discharged from the hospital. This study might help understand more about coronavirus and improve the awareness of pathogen detection in patients with coronavirus encephalitis. Keywords: coronavirus HKU1; encephalitis.

[The mechanism study of ouabain in inhibiting the growth of hepatocellular carcinoma cells by inhibiting the laser kinase signaling pathway].

Objective: To investigate the effect of Na(+)/K(+)-ATPase inhibitor ouabain on the proliferation and division of liver cancer HepG2 cells, and to explore the anticancer mechanism. Methods: HepG2 cells were exposed with different concentrations of ouabain (0.1, 1, 10 µmol/L) for 24 h, the proliferation ability was appraised using CCK-8, and the HepG2 cells was as a control group. The status of chromosome separation was detected with cell immunofluorescence (ICC) coupled to confocal microscope. The expression levels of AURKA, mTOR, p-mTOR, ERK and p-ERK protein were analyzed using western blot. Results: After treating with 0.1, 1 and 10 µmol/L of ouabain for 24 h, the inhibitory rate of cells were (23.5±4.57)%, (49.80±5.32)%, and (72.10±5.62)%, respectively. Ouabain could significantly inhibit the proliferation of HepG2, and presented in a dose-dependent manner(F=32.8, P<0.05). The ICC results showed that the chromosome separation disorders occurred in HepG2 cells treated with 1 µmol/L for 24 h, and the spindle diameter of HepG2 cells with ouabain treatment was decreased significantly compared with the control group(t=9.58, P<0.05). The results of western blot showed that the expression levels of AURKA, p-mTOR and p-ERK expressions in HepG2 cells treated with 1 µmol/L of ouabain were significantly decreased compared with the control group(F=16.26, 8.32, 33.59, P<0.05). Ouabain inhibited the growth of hepatocellular carcinoma cells in nude mice(F=370.20, P<0.05). Conclusion: Ouabain can induce chromosome division disorder and inhibit the proliferation in liver cancer HepG2 cells by inhibiting AURKA signaling pathway.

[Comparison on efficacy and safety of different bridging anticoagulation therapies in patients undergoing mechanical heart valve replacement surgery].

Objective: To evaluate the efficacy and safety of different bridging anticoagulant therapies in patients undergoing mechanical heart valve replacement (MHVR) surgery. Methods: Consecutive patients undergoing MHVR surgery from January 2018 to December 2018 in First Hospital of Lanzhou University were prospectively enrolled in this study. Patients were divided into unfractionated heparin (UFH) group and low molecular weight heparin (LMWH) group according to the postoperative bridging anticoagulation methods. Preoperative clinical data and postoperative related time and cost parameters, including drainage time, duration of stay in intensive care unit (ICU), postoperative time (interval from end of operation to discharge) and INR stabilization time (interval from start of bridge anticoagulation to INR value reaching the standard for 2 consecutive days) of all enrolled patients were collected, and all patients were followed up for 4 weeks and thromboembolic or bleeding events were analyzed. Multivariate logistic regression was used to determine the independent prognostic factors of thromboembolic or bleeding events after MHVR receiving various bridging anticoagulant therapies. Results: A total of 217 patients were included in the study, including 120 patients in the UFH group and 97 patients in the LMWH group. Stroke occurred in two patients in the UFH group, while no stroke event occurred in the LMWH group. The incidence of bleeding events was significantly higher (9.28%(9/97) vs. 1.67%(2/120), P=0.02), while the drainage time, duration of stay in ICU, postoperative time, INR stabilization time were all significantly shorter in LMWH group than in UFH group (all P<0.05). Multivariate logistic regression analysis showed that bridging anticoagulation therapies (OR=0.18, 95%CI 0.04-0.86, P=0.03), fibrinogen level (OR=1.99, 95%CI 1.16-3.41, P=0.01) and creatinine level (OR=1.05, 95%CI 1.01-1.08, P=0.04) were independent prognostic factors for bleeding events. Conclusion: LMWH use is associated with increased risk of bleeding events, but can significantly reduce the drainage time, duration of stay in ICU, postoperative time, INR stabilization time in patients post MHVR surgery.

[Comparative study of differentiation potential of mesenchymal stem cells derived from orofacial system into vascular endothelial cells].

OBJECTIVE: To compare the proliferation and capacity of differentiation to vascular endothelial cells and angiogenesis induction among stem cells from human exfoliated deciduous teeth (SHED), dental pulp stem cells (DPSC) and human bone marrow mesenchymal stem cells (BMSC) from orofacial bone. METHODS: SHED and DPSC were isolated from pulp tissue of the patients. BMSC were isolated from orthognathic or alveolar surgical sites. The surface markers of the cells were detected by flowcytometry. Cell counting kit-8 (CCK-8) assays were conducted to detect the proliferation ability of the cells. The cells were induced into endothelial cells with conditional medium and then the induced cells were cultured in Matrigel medium. The expression of angiogenesis-related genes such as platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1), vascular endothelial growth factor receptor 2 (VEGFR2) and von Willebrand Factor (vWF) were quantified by real-time PCR. The cells were cultured in chick embryo chorioallantoic membrane (CAM) and the vessels were counted after 5 days. RESULTS: The cell surface markers CD73, CD90, CD105 and CD146 of all the stem cells were positive, CD34 and CD45 were negative. The CD146 positive rate of SHED and DPSC was higher than that of BMSC. SHED had a higher proliferation rate than DPSC and BMSC. After angiogenic induction for 14 d, 3 kinds of cells emanated pseudopodia formed grid structure long vasculature in Matrigel media. The total length of tube formation of induced BMSC (7 759.7 µm) and SHED (7 734.3 µm) was higher than DPSC (5 541.0 µm). The meshes number of induced SHED (70.7) was higher than DPSC (60) and BMSC (53.7) in Matrigel medium. The expression of CD31, VEGFR2 and vWF genes of SHED were higher than those of BMSC and DPSC. VEGFR1 gene expression of BMSC was higher than that of the other groups, and SHED was higher than DPSC. The expression of VEGF showed no difference among the cells. No deference was showed between the effect of the stem cells and negative control on new formed vessels in CAM. The total length of vessels of SHED (30.4 mm) was higher than that of the negative control (20.9 mm) and BMSC (28.0 mm). CONCLUSION: SHED, DPSC and BMSC can differentiate into vascular endothelial cells. SHED showed a stronger angiogenesis differentiation and proliferation potential compared with DPSC and BMSC.

[Effect of iron overload on the level of carbohydrate antigen 199 in type 2 diabetes mellitus].

Objective: To investigate the effect of iron overload on tumor marker CA199 in patients with type 2 diabetes mellitus (T2DM). Methods: Between January 2017 and June 2018, three hundred and twenty-three hospitalized patients with T2DM in Department of Endocrinology, Tianjin Fourth Central Hospital were selected as diabetes group and another 100 healthy persons as control group. All the participants with major diseases and factors affecting serum ferritin and CA199 level were excluded. Serum ferritin and CA199 levels were measured by chemiluminescence assay. Patients with a ferritin level above the normal range were defined as iron overload group. According to serum ferritin level, 178 patients were divided into iron overload group and 145 into normal ferritin group in the diabetes group. The levels of CA199 were compared between iron overload group and normal ferritin group, and the correlation was analyzed. Results: The levels of serum ferritin [407.1 (346.7) µg/L vs 266.6 (201.1) µg/L in males, 184.7 (133.0) µg/L vs 77.4 (64.0) µg/L in females,both P<0.001] and CA199 [18.2 (19.3) µg/L vs 9.3 (9.7) µg/L, P<0.001] in diabetes group were significantly higher than those in control group. Serum CA199 levels in iron overload group were significantly higher than those in normal ferritin group (20.7 µg/L vs 15.0 µg/L, P<0.001; above intermediate value percent:53.9% vs 35.8%,P=0.001). CA199 was positively correlated with glycosylated hemoglobin (HbA1c) (r=0.280, P<0.001) and iron overload (r=0.210, P<0.001). Multiple linear regression analysis showed that iron overload (ß'=0.203, P=0.002) and HbA1c (ß(')'=0.202, P=0.001) had a significant effect on CA199 level. Conclusion: In T2DM patients, iron overload is an important factor affecting serum CA199 level.

[Expression of circular RNA hsa_circ_0014130 in lung adenocarcinoma cell lines and its effect on proliferation and invasion of lung adenocarcinoma cell line].

Objective: To investigate the function and mechanism of hsa_circ_0014130 in lung adenocarcinoma cell line and to find potential molecular inhibitors. Methods: The hsa_circ_0014130 expression level detection and overexpression and subtraction experiments were performed using common cell lines of lung cancer (PC9, H1299, A549, HCC827, and BEAS-2B). qPCR was used to verify the proliferation and invasion of lung cancer cells by MTS and invasion assay, and then the targeted microRNA was searched through the database. Western blot was used to detect the downstream signaling pathways, and finally the effect of small molecule inhibitors was investigated on proliferation and invasion of non-small cell lung cancer. Results: The expression level of hsa_circ_0014130 was up-regulated in the three cell lines, and both the overexpression plasmid and the subtractive siRNA were effectively transfected into the cells. Overexpression of hsa_circ_0014130 was able to promote the proliferation and invasion of tumor cells, and knockdown of hsa_circ_0014130 inhibited the proliferation and invasion of tumor cells. hsa_circ_0014130 was capable to target hsa-miR-566 to reduce its expression level and to inhibit epithelial-to-mesenchymal transition. The use of the small molecule inhibitor SB-431542 and simultaneous reduction of hsa_circ_0014130 significantly inhibited the proliferation and invasion of tumor cells. Conclusions: The hsa_circ_0014130 promotes the invasion and proliferation of lung cancer cells by targeting hsa-miR-566 to enhance the expression of TWIST1, and its expression level can be significantly inhibited by the small molecule inhibitor SB-431542, which significantly inhibits the proliferation and invasion of lung cancer cells. Therefore,hsa_circ_0014130 is a potential lung cancer treatment target.

[Comparison of the properties of CD146 positive and CD146 negative subpopulations of stem cells from human exfoliated deciduous teeth].

OBJECTIVE: Stem cells from human exfoliated teeth (SHED) were sorted by magnetically activated cell sorting (MACS) technique to obtain the CD146 positive and negative cell subpopulation. Then the biological characteristics of these subpopulations were compared to explore their specific application potential in tissue engineering. METHODS: In this study, freshly extracted deciduous teeth without any caries or dental pulp disease were obtained. SHED was isolated using enzyme digestion method and then sorted by MACS, CD146 positive cells and CD146 negative cells were obtained after cell sorting. The biological characteristics of the unsorted mixed cells, CD146 positive subpopulation and CD146 negative subpopulation were compared. The proliferation ability was detected through cell counting kit-8 (CCK-8) and colony-forming unit (CFU). After osteogenic induction, alizarin red staining was performed and the gene expression of osteogenic related markers was detected by quantitative real-time polymerase chain reaction(qPCR). After adipogenic induction, oil-red O staining was performed and the gene expression of adipogenic related markers was detected. After neurogenic differentiation induction, the expression of neural markers was detected by immunofluorescence and the gene expression of neural markers was detected by qPCR. RESULTS: SHED of the fifth passage was sorted by MACS. And the CD146 positive cell subpopulation and CD146 negative cell subpopulation were obtained. CCK8 assay showed that the proliferative tendency of the three cell groups was consistent, but the proliferation potential of CD146 positive and negative cell subpopulations was significantly lower than that of the unsorted cells. The colony forming rates of the unsorted mixed cell group, CD146 positive and negative populations were 28.6%±3%,17.1%±2.3% and 27.5%±2.5%, respectively. After 21 days of osteogenic induction, alizarin red staining and qPCR showed that the CD146 positive cell population had more mineralized nodule formation and expressed higher level of osteogenic related genes compared with the other two groups. After 21 days of adipogenic induction, oil red O staining and qPCR results showed that the CD146 negative subpopulation produced more lipid droplets and the expression of lipid related genes increased more significantly. After 14 days of neural induction, cell immunofluorescence and qPCR results showed that the unsorted mixed cell group and CD146 positive subpopulation expressed glial cell marker, and the expressions of neural precursor cells and neuronal marker increased significantly in negative subpopulation. CONCLUSION: The unsorted mixed cells showed better proliferative potential than CD146 positive and negative subpopulations. The CD146 positive subpopulation was most potent in osteogenic differentiation; it was more suitable for bone tissue engineering. The CD146 negative cells had stronger adipogenic differentiation potential than the other two cell groups; different subpopulations differed in neural differentiation.

[Effect of telmisartan on peroxisome proliferator-activated receptor γ signaling pathway in 3T3-L1 adipocytes].

Objective: To investigate the mechanisms of telmisartan on delaying the course of type 2 diabetes. Methods: 3T3-L1 preadipocytes were induced to 80% mature adipocytes (control group) and stimulated with 50 ng/ml tumor necrosis factor (TNF-α) for 1 hour (TNF-α group). Then 0.1, 5, 10 µmol/L telmisartan was added to the culture medium for 24 h, respectively(T(0.1, )T(5) and T(10) group). The cells from each group was collected to detect peroxisome proliferator-activated receptors γ (PPARγ) and its phosphorylation level, as well as upstream kinase cell cycle dependent kinase 5 (CDK5) by Western blot. Adiponectin in the culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). PPARγ or CDK5 of undifferentiated 3T3-L1 adipocytes were silenced by using targeted short hairpin RNA (shRNA). Through infection of the cells with the retrovirus, stabled PPARγ or CDK5 knockdown cell lines were set up and screened by incubation with puromycin. 3T3-L1 adipocyte cell lines expressing serine mutant PPARγ (S273A, S112A, S186A) were obtained and thus their phosphorate sites were further determined. CDK5 knockdown cell lines were detected by oil red O staining to measure the lipid accumulation and differentiation efficiency. The 10 µmol/L telmisartan was used to treat mature CDK5 knockdown 3T3-L1 adipocytes, Western blot was used to detect PPARγ and its phosphorylation level, and ELISA was used to detect the release of adiponectin in the culture supernatant. Results: The TNF-α stimulation had no significant effect on the expression of PPARγ in each group (all P>0.05), but it could up-regulate the phosphorylation of PPARγ in the TNF-α group and down-regulate the release of adiponectin (all P<0.05). Compared with TNF-α group, telmisartan can reduce PPARγ phosphorylation levels and up-regulate adiponectin release in different degrees, among which T(5) group and T(10) group had statistically significant differences (all P<0.05), but for T(0.1) group, the difference was not significant (P>0.05). Compared with the 3T3-L1 wild type (WT) adipocytes, adiponectin in cell line with only S273A mutant did not respond to TNF-α stimulation and telmisartan intervention. Oil red O staining showed that silencing of CDK5 did not affect the differentiation of 3T3-L1 preadipocytes. Western blot results showed that silencing of CDK5 (shCDK5 group) had no significant effect on PPARγ expression (P>0.05), but it could down-regulate the phosphorylation of PPARγ and up-regulate release of adiponectin, compared with the randomized control group (shCon group) and the differences were statistically significant (all P<0.05). Conclusions: Telmisartan can alleviate the increased PPARγ phosphorylation and up-regulation of adiponectin content due to TNF-α stimulation. CDK5 mediates the effect of telmisartan on PPARγ signaling pathway in 3T3-L1 adipocytes. Additonally, it also demonstrated the action site of telmisartan was PPARγ Ser 273, and CDK5 is upstream kinases of PPARγ.

[Effect of different parts of skull thickness on stereotactic electroencephalogram in children].

Objective: To investigate the thickness of cranial bone in different parts of children skull during stereotactic electroencephalogram (SEEG) and its effect on electrode fixation. Methods: From October 2016 to March 2017, 13 children with SEEG by robot of surgery assistant (ROSA) were selected. The basic case information and electrode design scheme were collected. The skull thickness of each electrode channel was measured on post-operation CT, and the loosening of the fixed screws were recorded. The thickness of skull in frontal bone, temporal bone, parietal bone and occipital bone was statistically processed by SPSS statistical software. Results: There were total 113 electrodes in 13 children with epilepsy. There were 45 electrodes at frontal bone, of which the thickness was (5.7±2.8)mm. There were 34 electrodes at temporal bone, of which the thickness was (3.5±1.3)mm.There were 16 electrodes at parietal bone, of which the thickness was (6.0±2.5)mm.There were 18 electrodes at occipital bone, of which the thickness was (6.9±0.5)mm. Statistics showed that there was significant difference between differnt bone (F=15.340, P<0.01). There were 4 electrodes loosening, 1 at frontal bone and 3 at temporal bone, when the screws were removed. There was no adverse event related to the implantation of electrodes. Conclusions: The children's skull thickness is thinner than adults. The screw loosening is exist in some cases, but it has no effect on SEEG recording. No SEEG related adverse events are found in this group. Therefore, ROSA guided SEEG is safe and reliable in children with epilepsy.

Combined multiplex loop-mediated isothermal amplification with lateral flow assay to detect sea and seb genes of enterotoxic Staphylococcus aureus.

UNLABELLED: Staphylococcal enterotoxins (SEs) are the most common cause of food poisoning worldwide and can induce symptoms, such as diarrhoea, vomiting and abdominal cramping. Thus, the aim of this study is to develop a multiplex loop-mediated isothermal amplification combined with a lateral flow assay (m-LAMP/LFA) to simultaneously detect the sea and seb genes of Staphylococcus aureus. The amplicons of the sea gene were labelled with digoxigenin (Dig) and biotin while those of seb gene were labelled with fluorescein isothiocyanate (FITC) and biotin. These amplicons were detected using a multiplex LFA with NeutrAvidin-tagged gold nanoparticles as the detection reagent. After optimization, the detection limit of this assay was 10(2)  CFU ml(-1) Staph. aureus, which was one tenth that of a multiplex PCR. This assay did not exhibit any cross-reactivity in detecting other enterotoxic Staph. aureus strains or other food pathogens. After 6 h of enrichment, this developed assay detected 1 CFU ml(-1) of Staph. aureus in milk, apple juice, cheese and rice. The developed m-LAMP/LFA method does not require expensive equipment and can be completely implemented within an 8-h workday. Therefore, this method can provide an effective means of quickly screening staphylococcal enterotoxin A- and/or staphylococcal enterotoxin B-producing Staph. aureus in food samples. SIGNIFICANCE AND IMPACT OF THE STUDY: Staphylococcus aureus is one of the major foodborne pathogens worldwide, and its staphylococcal enterotoxin A and B are strongly associated with food poisoning. This work developed a multiplex loop-mediated isothermal amplification combined with a lateral flow assay (m-LAMP/LFA) to simultaneously detect the sea and seb genes of Staph. aureus in food samples. The assay has good specificity and sensitivity with ease-of-use features, making it ideal for on-site detection.

Utility of dysphagia grade in predicting endoscopic ultrasound T-stage of non-metastatic esophageal cancer.

Patients with non-metastatic esophageal cancer routinely undergo endoscopic ultrasound (EUS) for loco-regional staging. Neoadjuvant therapy is recommended for ≥T3 tumors while upfront surgery can be considered for ≤T2 lesions. The aim of this study was to determine if the degree of dysphagia can predict the EUS T-stage of esophageal cancer. One hundred eleven consecutive patients with non-metastatic esophageal cancer were retrospectively reviewed from a database. Prior to EUS, patients' dysphagia grade was recorded. Correlation between dysphagia grade and EUS T-stage, especially in reference to predicting ≥T3 stage, was determined. The correlation of dysphagia grade with EUS T-stage (Kendall's tau coefficient) was 0.49 (P < 0.001) for the lower and 0.59 (P = 0.008) for the middle esophagus. The sensitivity and specificity of dysphagia grade ≥2 (can only swallow semi-solids/liquids) for T3 cancer were 56% (95% confidence interval [CI] 43-67%) and 93% (95% CI 79-98%), respectively. The sensitivity, specificity, and positive predictive value of dysphagia grade ≥3 (can only swallow liquids or total dysphagia) for T3 lesions were 36% (95% CI 25-48%), 100% (95% CI 89-100%), and 100% (95% CI 83-100%), respectively. Overall, there was a significant positive correlation between dysphagia grade and the EUS T-stage of esophageal cancer. All patients with dysphagia grade ≥3 had T3 lesions. This may have clinical implications for patients who can only swallow liquids or have complete dysphagia by allowing for prompt initiation of neoadjuvant therapy, especially in countries/centers where EUS service is difficult to access in a timely manner or not available.