Stringent test of QED with hydrogen-like tin.

Inner-shell electrons naturally sense the electric field close to the nucleus, which can reach extreme values beyond 1015 V cm-1 for the innermost electrons1. Especially in few-electron, highly charged ions, the interaction with the electromagnetic fields can be accurately calculated within quantum electrodynamics (QED), rendering these ions good candidates to test the validity of QED in strong fields. Consequently, their Lamb shifts were intensively studied in the past several decades2,3. Another approach is the measurement of gyromagnetic factors (g factors) in highly charged ions4-7. However, so far, either experimental accuracy or small field strength in low-Z ions5,6 limited the stringency of these QED tests. Here we report on our high-precision, high-field test of QED in hydrogen-like 118Sn49+. The highly charged ions were produced with the Heidelberg electron beam ion trap (EBIT)8 and injected into the ALPHATRAP Penning-trap setup9, in which the bound-electron g factor was measured with a precision of 0.5 parts per billion (ppb). For comparison, we present state-of-the-art theory calculations, which together test the underlying QED to about 0.012%, yielding a stringent test in the strong-field regime. With this measurement, we challenge the best tests by means of the Lamb shift and, with anticipated advances in the g-factor theory, surpass them by more than an order of magnitude.

[Association of urinary cadmium level with body mass index and body circumferences among older adults over 65 years old in 9 longevity areas of China].

Objective: To investigate the association of urinary cadmium level with body mass index (BMI) and body circumferences among the older adults over 65 years old in 9 longevity areas of China. Methods: Subjects were older adults over 65 years old from the Healthy Aging and Biomarkers Cohort Study (HABCS) between 2017 and 2018 conducted in 9 longevity areas in China. A total of 1 968 older adults were included in this study. Information including socio-demographic characteristics, lifestyles, diet intake, and health status was collected by using questionnaires and physical examinations. Urine samples were collected to detect urinary cadmium and creatinine levels. Body circumferences included waist circumference, hip circumference and calf circumference. Subjects were divided into three groups (low:<0.77 µg/g·creatinine, middle:0.77-1.69 µg/g·creatinine, high:≥1.69 µg/g·creatinine) by tertiles of creatinine-adjusted urinary cadmium concentration. Multiple linear regression models were used to analyze the association of creatinine-adjusted urinary cadmium level with BMI and body circumferences. The dose-response relationship of creatinine-adjusted urinary cadmium concentration with BMI and body circumferences was analyzed by using restrictive cubic splines fitting multiple linear regression model. Results: The mean age of subjects was (83.34±11.14) years old. The median (Q1, Q3) concentration of creatinine-adjusted urinary cadmium was 1.13 (0.63, 2.09) µg/g·creatinine, and the BMI was (22.70±3.82) kg/m2. The mean values of waist circumference, hip circumference, and calf circumference were (85.42±10.68) cm, (92.67±8.90) cm, and (31.08±4.76) cm, respectively. After controlling confounding factors, the results of the multiple linear regression model showed that for each increment of 1 µg/g·creatinine in creatinine-adjusted urinary cadmium, the change of BMI, waist circumference, hip circumference, and calf circumference in the high-level group was -0.28 (-0.37, -0.19) kg/m2, -0.74 (-0.96, -0.52) cm, -0.78 (-0.96, -0.61) cm, and -0.20 (-0.30, -0.11) cm, respectively. The restrictive cubic splines curve showed a negative nonlinear association of creatinine-adjusted urinary cadmium with BMI (Pnonlinear<0.001) and negative linear associations of creatinine-adjusted urinary cadmium with waist circumference (Plinear<0.001), hip circumference (Plinear<0.001), and calf circumference (Plinear<0.001). Conclusion: Urinary cadmium level is significantly associated with decreased BMI, waist circumference, hip circumference and calf circumference among older adults over 65 years old in 9 longevity areas of China.

[Synchronization isolation method for multiple types of cells from mouse liver].

Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.

[Association of small dense low-density lipoprotein cholesterol with H-type hypertension and MTHFR gene polymorphism].

To investigate the expression of small dense low-density lipoprotein cholesterol (sdLDL-C) in patients with H-type hypertension and its association with H-type hypertension and methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms. The retrospective study method was used,and a total of 207 hospitalized hypertensive patients (76 males and 131 females, aged 40-82 years, median age 66 years) admitted to the Zibo First Hospital from March 2021 to March 2022 were enrolled in this study. The levels of homocysteine (Hcy), sdLDL-C, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), total cholesterol (TC), triglyceride (TG) and lipoprotein (a) [Lp(a)] were measured. The patients were divided into H-type hypertensive group (n=105, 40 males and 65 females) and non-H-type hypertensive group (n=102, 36 males and 66 females) according to Hcy levels. The C677T polymorphism of methylenetetrahydrofolate reductase (MTHFR) gene was detected in each group. Logistic regression analysis was performed for the related factors of H-type hypertension. The serum sdLDL-C levels were (0.92±0.31) and (0.65±0.28) mmol/L in H-type hypertension group and non-H-type hypertension group, respectively. The sdLDL-C levels in H-type hypertension group were significantly higher than those in non-H-type hypertension group (t=6.500, P<0.01). There was no significant difference in the serum sdLDL-C levels between males and females in H-type hypertension group (t=-1.543, P=0.129). The CC, CT, TT genotypes and C and T allele frequencies of MTHFR C677T in H-type hypertension group were significantly different from those in non-H-type hypertension group (P<0.05). The Hcy and sdLDL-C levels in different genotypes of MTHFR in H-type hypertension group were significantly different (H=12.742, P=0.002; F=3.345, P=0.042). Among them, Hcy levels were higher in TT genotype than in CT and CC genotypes, respectively (Z=-28.099, P=0.003; Z=-16.112, P=0.040), and sdLDL-C levels were higher in TT genotype than in CC genotype (t=-2.587, P=0.012). Logistic regression analysis showed that age, sdLDL-C, and MTHFRC677T TT genotypes were associated with the development for H-type hypertension. In conclusion, the level of sdLDL-C is associated with MTHFR gene polymorphisms and may be associated with the development of H-type hypertension.

[Characteristic analysis of patients with type 2 diabetes mellitus combined with advanced stage chronic liver disease: a community population-based cross-sectional study in Heping District, Shenyang City (CHESS-LN 2101)].

Objective: To investigate the prevalence and independent risk factors of non-alcoholic fatty liver disease (NAFLD) and advanced chronic liver disease among the type 2 diabetes mellitus (T2DM) population in the Shenyang community, so as to provide evidence for the prevention and control of T2DM combined with NAFLD. Methods: This cross-sectional study was conducted in July 2021. 644 T2DM cases from 13 communities in Heping District, Shenyang City were selected. All the surveyed subjects underwent physical examination (measurements of height, body mass index, neck circumference, waist circumference, abdominal circumference, hip circumference, and blood pressure), infection screening (excluding hepatitis B and C, AIDS, and syphilis), random fingertip blood glucose, controlled attenuation parameter (CAP), and liver stiffness measurement (LSM). The study subjects were divided into the non-advanced chronic liver disease group and the advanced chronic liver disease group according to whether the LSM value was greater than 10 kPa. Cirrhotic portal hypertension development was indicated in patients with LSM ≥ 15 kPa. The comparison of multiple mean values among the sample groups was performed by analysis of variance when the normal distribution was met. Results: In the T2DM community population, there were 401 cases (62.27%) combined with NAFLD, 63 cases (9.78%) combined with advanced chronic liver disease, and 14 cases (2.17%) combined with portal hypertension. There were 581 cases in the non-advanced chronic liver disease group and 63 cases (9.78%) in the advanced chronic liver disease group (LSM ≥10 kPa), including 49 cases (7.61%) with 10 kPa≤LSM<15 kPa, 11 cases (1.71%) with 15 kPa ≤LSM<25 kPa, and 3 cases (0.47%) with LSM ≥ 25 kPa. Age, body mass, body mass index, neck circumference, waist circumference, hip circumference, waist-to-height ratio, systolic blood pressure, and CAP were all statistically different between the non-advanced chronic liver disease group and the advanced chronic liver disease group (F=-1.983,-2.598,-4.091,-2.062,-3.909, -4.581,-4.295,-2.474, and -5.191, respectively; P<0.05). There was a statistically significant difference in terms of whether or not there was combined cerebrovascular disease (2=4.632, P=0.031); however, there were no statistically significant differences in terms of lifestyle, diabetes complications, and other complications (P>0.05). Conclusion: Patients with T2DM have a higher prevalence of NAFLD (62.27%) than those with advanced chronic liver disease (9.78%). 2.17% of T2DM cases in the community may not have had early diagnosis and early intervention, and they might have been combined with cirrhotic portal hypertension. So, the management of these patients should be strengthened.

Direct Q-Value Determination of the ß

The cyclotron frequency ratio of ^{187}Os^{29+} to ^{187}Re^{29+} ions was measured with the Penning-trap mass spectrometer PENTATRAP. The achieved result of R=1.000 000 013 882(5) is to date the most precise such measurement performed on ions. Furthermore, the total binding-energy difference of the 29 missing electrons in Re and Os was calculated by relativistic multiconfiguration methods, yielding the value of ΔE=53.5(10) eV. Finally, using the achieved results, the mass difference between neutral ^{187}Re and ^{187}Os, i.e., the Q value of the ß^{-} decay of ^{187}Re, is determined to be 2470.9(13) eV.

[Analysis of the relationship between the anatomical location of intrapulmonary metastatic lymph nodes and relapse risk and survival in patients with N1 non-small cell lung cancer].

Objective: To evaluate the relationship between the anatomical location of intrapulmonary metastatic lymph nodes and relapse risk and survival in patients with N1 non-small cell lung cancer(NSCLC). Methods: A retrospective analysis of the clinical and pathological data of 138 patients with completely resected N1 NSCLC was conducted. There were 79 males and 59 females, aged from 26 to 81 years with an average of (59±10) years. All of them were treated in the Department of Thoracic Surgery Ⅱ of Peking University Cancer Hospital between January 2007 and December 2015. Patients were stratified based on the 8th edition of the American Joint Committee on Cancer (AJCC) N1 classification and the modified pathological N1 classification strategy, respectively. According to modified pathological N1 classification strategy, which was defined based on the anatomical location of intrapulmonary metastatic lymph nodes, N1 nodes were subcategorized into the hilar (stations 10-11, mN1b) (n=36) and peripheral (stations 12-14, mN1a) (n=102) zones. The Kaplan-Meier curves were plotted to compare the relapse risk and survival analysis, disease-free survival (DFS), and overall survival (OS) were compared between the two staging methods through univariate and multivariate analysis to evaluate the effectiveness of the two classifications in stratifying patients with distinct risks of disease relapse and survival. Results: According to the modified N1 classification, the differences in 5-year DFS and OS between the subgroups (mN1a vs mN1b) were statistically significant(59.5% vs 35.7%; 81.2% vs 56.0%; both P<0.05). However, following the 8th edition of the AJCC N1 classification, no significant differences were found in DFS and OS between the subgroups (both P>0.05). Multivariate analysis showed that the modified N1 classification was an independent prognostic factor to DFS (HR=1.814, 95%CI: 1.005-3.275) and OS (HR=3.919, 95%CI: 1.918-8.009) (all P<0.05). However, the 8th edition of the AJCC N1 classification was not an independent prognostic factor to DFS (HR=1.360, 95%CI:0.767-2.412) or OS (HR=1.620, 95%CI:0.839-3.131) (both P>0.05) as revealed by multivariate analysis. Conclusions: The relapse risk and survival could be assessed effectively using the modified pathological N1 classification, which was defined and subcategorized based on the anatomical location of intrapulmonary metastatic lymph nodes for N1 NSCLC patients. The modified pathological N1 classification is superior to the 8th edition of the AJCC classification.

[Study on maternal-fetal status of Pb, Cd, As, Hg, Mn and Se elements and transplacental transfer efficiency].

Objective: To understand the levels of Pb, Cd, As, Hg, Mn, and Se in maternal and umbilical cord blood, and to explore the transplacental transfer efficiency (TTE). Methods: From September 2010 to December 2013, a total of 773 pregnant women and their newborns (Laizhou Bay Birth Cohort) were recruited from a second grade hospital in the south bank of Laizhou Bay, Bohai, Shandong Province. According to different detection methods, the six measured elements are classified into three groups including the Hg measurement group (595 mother-newborn pairs), the Pb measurement group (534 mother-newborn pairs), and the Cd, As, Mn and Se measurement group (244 mother-newborn pairs). The demographic characteristics of pregnant women and their newborns were obtained by the questionnaire. The concentrations of elements in maternal and umbilical cord blood were detected and the TTE of each element (elemental concentration in cord blood/elemental concentration in maternal blood) was calculated. The correlation of elements between maternal and cord blood was analyzed using Spearman's rank correlation coefficient. Results: The mean±SD of maternal age, gestational week and newborn birth weight of 773 mother-infant pairs were (28.34±4.50) years, (39.47±1.39) weeks and (3 419.47±497.39) g respectively. The median concentrations of Pb, Cd, As, Hg, Mn and As in maternal and cord blood were 31.12 and 30.02, 1.19 and 0.47, 8.05 and 6.03, 0.69 and 1.26, 100.70 and 105.55, 127.25 and 115.00 µg/L, respectively. The TTE of Pb, Cd, As, Hg, Mn, and Se was 0.98, 0.41, 0.73, 1.73, 0.96 and 0.91, respectively. Pb, Cd, Hg, Mn, and Se showed a significant positive correlation between maternal blood and cord blood, with Spearman correlation coefficients of 0.397, 0.298, 0.698, 0.555, and 0.285 (all P values<0.001). Conclusion: Each element was commonly detected in maternal blood and cord blood. The TTE of Hg was the highest.

[Establishment of the method for measuring upper airway critical closing pressure].

Objective: To establish a noninvasive method for measuring upper airway critical closing pressure (Pcrit), so as to evaluate collapsibility of the upper airway during sleep. Methods: Pcrit was determined through the use of a noninvasive positive/negative pressure (CPAP/CPNP) ventilator(with independent intellectual property rights) during stageⅡ of non-rapid eye movement sleep. For the direct measurement, Pcrit was the pressure below which the upper airway occluded. For the indirect measurement, nasal pressure was plotted against maximum inspiratory flow (Vimax), and linear regression was used to interpolate the pressure (i.e., Pcrit) at which zero flow occurred. Pcrit was attained from 19 subjects without obstructive sleep apnea syndrome(OSAS), and the correlation between direct and indirect measurement methods was analyzed. Results: Directly measured and indirectly measured Pcrit showed no significant difference [(-7.02±2.74 vs (-7.26±2.96) cmH2O, 1 cmH2O=0.098 kPa; t=1.667, P>0.05] and had a highly significant correlation (r=0.986, P=0.000). Bland-Altman analysis revealed that the mean between-method difference was (0.24±0.53) cmH2O, and 95% limits of agreement ranged from -0.80 to 1.27 cmH2O, and all points except one were within limits of agreement. Conclusion: Pcrit derived from the direct and indirect measurement methods does not differ, and both methods could be used for evaluating the upper airway collapsibility.

[Application of a small drainage tube in transoral endoscopic thyroidectomy vestibular approach].

Objective: To examine the feasibility of small drain in transoral endoscopic thyroidectomy vestibular approach (TOETVA). Methods: A prospective research was performed in Department of Oncology Plastic Surgery/Head and Neck Surgery, Hunan Cancer Hospital, from October 2018 to March 2019. Totally 103 patients who met the inclusion and exclusion criteria, signed the operation agreement of TOETVA, had their surgery completed and followed-up over 6 months, were enrolled in analysis. A central venous catheter was used as a drain tube in all cases (outer diameter 1.7 mm, inner diameter 1.0 mm). Visual analogue scale (VAS) was applied for assessing pain scores during the first 24 hours after the operation. Vancouver scar scale (VSS) was used for assessing the scar left by the drainage. The drainage volume (minimum scale:10 ml, approximate read: 1 ml) was recorded every 2 hours during the first postoperative 12 hours, every 4 hours during 12 to 24 hours, every 8 hours during 24 to 48 hours, and once from 48 hours until extubation. The volume of drainage, the cumulative volume and the percentage of cumulative volume accounting for the total volume were calculated. The data of residual volume (subtract the cumulative volume from the total volume) in the postoperative 24, 32 and 40 hours were analyzed, and their upper one-side P(95) was calculated by percentile method. Results: There were 12 males and 91 females. The age was (36.6±9.7) years (range: 18 to 58 years). The intraoperative tube-inserting time was (10.1±2.6) minutes (range: 6 to 18 minutes). The pain score on the first day was 2.7±1.1 (range: 1 to 5). The extubation time was (2.7±0.5) days (range: 2 to 4 days). VSS scores in the postoperative 1(st) month and 6(th) month were 2.9±1.3 (range: 0 to 7) and 0(2)(M(Q(R))), respectively. The size of the scar was 0 (2.5) mm in the postoperative 6(th) month. Sixty-four patients had no visible scars. There were 5 patients who had postoperative drain-related complications (1 for air leakage, 1 for tube blockage, 3 for subcutaneous hydrop, 2 for regional infection), who were all cured after proper treatment. The total volume of drainage for 98 patients without postoperative drain-related complications was (80.1±12.2) ml (range: 58 to 131 ml). The cumulative drainage within 8 hours accounted for (53.8±4.2)% (range: 41.0% to 62.9%) of the total drainage. The volume of residual fluids in the postoperative 32 hours was estimated to (5.8±2.7) ml (range: 0 to 12 ml,P(95)=10.0 ml). Conclusions: The small drain tubecan be applied in TOETVA, providing a satisfied cosmetic appearance and a reliable drainage. The main exudation period of the wound is within 8 hours after the operation. If a residual volume less than 10 ml is considered to be self-absorbable, the shortest safe extubation point for 95% patients without drain-related complications should be 32 hours after the operation.

CCR6 defines a subset of activated memory T cells of Th17 potential in immune thrombocytopenia.

Current researches have determined the significance of C-C chemokine receptor (CCR)6 expression as either a marker of T helper cells (Th) or an effector and regulator of T cell function. However, the roles of CCR6 in the pathogenesis of immune thrombocytopenia (ITP) are unclear. In this study, we aimed to investigate the phenotype and functional characteristics of circulating CCR6+ T cells in blood from chronic ITP patients and healthy controls. We found that the frequency of CCR6+ CD4+ cells was higher in ITP patients than in healthy controls. Anti-CD3/anti-CD28 stimulation induced rapid expansion of CCR6+ CD4+ cells in ITP patients. CCR6+ CD4+ cells had a phenotype of activated cells and predominantly expressed CD45RO. Forkhead box protein P3 (FoxP3) and CD25-positive cells were exclusively detected within the CCR6+ CD4+ cells. In ITP patients, CCR6+ regulatory T cells (Treg ) were decreased and positively correlated with platelet counts and transforming growth factor (TGF)-ß plasma levels. In contrast to CCR6- counterparts, CCR6+ CD4+ cells produced higher levels of interleukin (IL)-17A. The frequency of CCR6+ Th17 was higher in ITP patients and positively correlated with IL-17A levels in supernatant. Most importantly, CCR6+ CD4+ cell subpopulations, but not CCR6- CD4+ , were closely correlated to treatment response of ITP patients. These findings suggest that circulating CCR6+ CD4+ cells in ITP patients have characteristics of activated memory Th17 phenotype and could be used to monitor disease activity and treatment response.

[The blood lead level and its relationship with behavior in school-age children from rural areas of Chongqing].

Objective: To explore the blood lead level and its relationship with behavior in school-age children from rural areas of Chongqing. Methods: A total of 697 students from grades 3 to 6 in the fall semester of 2014 from 14 rural townships in one district of Chongqing was selected by using the random cluster sampling method. Blood were sampled to analyze the lead level. Neurobehavioral tests were performed to determine their personal cognitive and memory ability. Questionnaires and physical examinations were administered to obtain the information of confounding factors. All students were divided into Q1-Q4 groups according to the quartile of their blood lead level. The relationship between the blood lead level and behavior was analyzed by multivariate logistic regression model and restricted spline regression model. Results: The mean age of 697 students was (10.07±1.36) years old, and the median (interquartile range) of their blood lead level was 44.31 (35.42) µg/L. Multivariate logistic regression model showed that after adjusting for age, gender, body mass index and maternal culture level, compared with Q1 group, the OR (95%CI) values of high digit symbol substitution test (DSST) scores and high overall memory quotient (MQ) scores in Q3 group were 1.65 (1.01-2.70) and 2.10 (1.21-3.62), and the OR (95%CI) value of high long term memory (LTM) scores in Q4 group was 0.53 (0.31-0.92). The results of the restricted spline regression model showed that the dose-response curves between the blood lead level and MQ/LTM test scores were both parabolic (P<0.05). Conclusion: The blood lead level of school-age children from rural areas of Chongqing is the same as that from other areas of China, but slightly higher than that from other areas of Chongqing. Children with higher blood lead level have poor long-term memory ability.

[The clinical applied analysis of the modified transoral endoscopic thyroidectomy vestibular approach].

Objective: To investigate the safety and feasibility of the modified transoral endoscopic thyroidectomy vestibular approach (TOETVA) by dissection of mental nerve in clinical practice. Methods: Totally 140 patients underwent the modified TOETVA from the Department of Head and Neck Surgery, Hunan Cancer Hospital from July 2016 to June 2018 were analyzed retrospectively. There were 130 females and 10 males, aging (35.4±9.8) years (range: 11 to 56 years). The operative time, intraoperative blood loss, postoperative hospital stay, postoperative suction drainage, postoperative pain score, postoperative cosmetic satisfaction and postoperative complications (recurrent laryngeal nerve palsy, hypoparathyroidism, infection, pneumoderm, seroma and mental nerve injury) were summarized. Results: Of the 140 patients, 1 patient was transferred to open surgery. Fifty-nine patients underwent thyroidectomy with an operation time of (100.8±18.9) minutes. Sixty-three patients underwent thyroidectomy and central lymphadenectomy with an operation time of (112.1±16.6) minutes. Eighteen cases underwent total thyroidectomy and central lymphadenectomy with an operation time of (185.3±25.9) minutes. The postoperative hospital stay was (3.76±0.98) days. The postoperative drainage was (96.8±36.2) ml. The 24-hour postoperative pain score was 2.66±1.23, the postoperative cosmetic satisfaction was 9.65±0.24. Among the postoperative complications, there were 3 cases of temporary recurrent laryngeal nerve palsy, 2 cases of permanent recurrent laryngeal nerve palsy, 4 cases of temporary hypoparathyroidism but no permanent hypoparathyroidism, 2 cases of infection, 1 case of seroma, 3 cases of pneumoderm, and no cases of mental nerve injury. Conclusion: The modified TOETVA by dissection of mental nerve is safe and feasible.

Prevalence of type 5 familial hemophagocytic lymphohistiocytosis in Korea and novel mutations in STXBP2.

Familial hemophagocytic lymphohistiocytosis (F-HLH or FHL) is a potentially fatal immune dysregulation syndrome with a heterogeneous genetic background. Most recently, STXBP2 has been identified as the causative gene of type 5 FHL (FHL5) with a worldwide distribution. In this study, we investigated the prevalence of FHL5 in Korea. About 50 Korean pediatric patients with HLH who lacked pathogenic mutations in PRF1, UNC13D, or in STX11 from the previous series of 72 patients with HLH were analyzed for STXBP2 mutations by conventional sequencing analyses. As a result, we found one patient with two novel mutations of STXBP2: c.184A>G and c.577A>C. c.184A>G (p.Asn62Asp) was located within a highly conserved region of the STXBP2 protein and predicted to be deleterious. c.577A>C in exon 7 resulted in incomplete splicing mutation with exon 7 skipping concurrent with exon 7-retained transcript with p.Lys193Gln substitution. The frequency of FHL5 was ~1% (1/72) in Korean pediatric patients with HLH. This is the first study on FHL5 in Korea, and the data from a nationwide patient cohort provide another piece of genetic profiles of FHL.

Identification of mutations in the F8 and F9 gene in families with haemophilia using targeted high-throughput sequencing.

INTRODUCTION: At present, many methods are available for the genetic diagnosis of haemophilia, including indirect linkage analysis, direct sequencing. However, these methods are time-consuming, labourious, and limited in their application. Therefore, the development of new, more effective techniques is necessary. AIM: To detect the F8 and F9 gene mutations in patients with haemophilia and their female relatives in 29 haemophilia A (HA) and 11 haemophilia B (HB) families. FVIII: C and FIX:C were analyzed using one-stage method, and factor VIII and factor IX inhibitors were tested using the Bethesda method. Intron 22 and one inversions were identified using long-distance polymerase chain reaction (PCR) and standard PCR. Non-inversion mutations of the F8 and F9 gene were identified by targeted high-throughput sequencing. All mutations were verified by Sanger sequencing. RESULTS: Intron 22 inversion was detected in eight HA families and intron one inversion was detected in one HA family. Apart from the inversion mutations, 20 mutations were identified in HA families, including 17 previously reported and three novel mutations: c.5724G>A (p.Trp1908*), c.6116-1_6120delGAGTGTinsTCC (p.Lys2039Ilefs*13), and c.5220-2A>C. We found a complex rearrangement in HA: intron one inversion concomitant with exon one deletion. In HB, eight recurrent mutations were detected, including six missense mutations and two nonsense mutations. CONCLUSION: Targeted high-throughput sequencing is an effective technique to detect the F8 and F9 gene mutations, especially for the discovery of novel mutations. The method is highly accurate, time-saving and shows great advantage in uncovering large deletion mutations and also in distinguishing the wild-type genotype and heterozygous large deletions.

[Impact of biological function on ovarian clear cell carcinoma ES2 cell line with ARID1A gene expression down-regulating in vitro].

OBJECTIVE: To investigate the efficiency of biological function of AT rich interaction domain 1A (ARID1A) gene silenced by small interfering RNA (siRNA) on ovarian clear cell carcinoma ES2 cell line. METHODS: (1) The three pairs ARID1A gene siRNA interference fragments siN1 (ARID1A-705), siN2 (ARID1A-1513), siN3 (ARID1A-2282) and one pair negative control were respectively designed, and transfected into ES2 cells by RNA interference max reagent transiently. Reverse transcription (RT)-PCR and western blot methods were used to detect the expression of ARID1A mRNA and protein in ES2 cells transfected with interference fragments respectively. So as to select the best silencing effect of siRNA interference fragment(that was siN3), and then was used in the following experiment. (2) The following experiment were divided into three groups, namely siN3 transfection group, negative control group and blank control group. The proliferative activity of three groups of cells after transient transfection (6, 24, 48, 72, 96 hours) was assessed by cell counting kit-8 (CCK-8) assay and expressed as absorbance (A) value; the apoptosis rate of three groups of cells transfected transiently with interference fragment was measured by flow cytometry with annexin V/propidium iodide (PI) staining; the ability of cellular invasion of three groups of cells transfected transiently with interference fragment was tested by transwell experiment; the expression of nuclear factor-kappa B (NF-κB), membrane type-1 matrix metalloproteinase (MT1-MMP) and matrix metalloproteinase-2 (MMP2) protein in ES2 cells transfected transiently with interference fragment was measured by western blot. RESULTS: (1) The RT-PCR results showed that the ARID1A mRNA relative expression levels in ES2 cells after transfected transiently with siN1, siN2 and siN3 were 0.007 8±0.005 7, 0.006 8±0.000 3 and 0.002 8±0.000 3 respectively. They were all apparently lower than that in the negative control group (0.034 6±0.001 3; all P<0.01). The western blot results showed that the expression levels of ARID1A protein were 0.439 4±0.000 7, 0.424 4±0.005 0 and 0.386 0±0.005 8 respectively. They were also lower than that in the negative control group (0.732 4 ±0.030 3; all P<0.01). The siN3 with the highest transfection efficiency was selected to use in the following experiment. (2) The CCK-8 method showed that the proliferative activity of siN3 transfection group cells after transfected transiently at 6 hours was not statistically significant difference compared with those in negative control group and blank control group (0.506±0.010, 0.491±0.006, 0.498±0.009, respectively; all P>0.05). However, the proliferative activity of siN3 transfection group cells after transfected transiently at 24, 48, 72, 96 hours were higher than those in negative control group and blank control group (all P<0.01). The flow cytometry results showed that the apoptosis rate of siN3 transfection group cells was (20.0±3.9)%, which was significantly lower than those in negative control group and blank control group [(31.5±5.0)%, (34.0±4.2)%, respectively; all P<0.05]. The transwell experiment showed that the penetrated cell counts of siN3 transfection group was 60.4±2.9, which was apparently higher than those in negative control group and blank control group (54.2±3.5, 52.1±3.8, respectively; all P<0.01). Western blot experiment showed that the relative expression levels of NF-κB, MT1-MMP and MMP2 protein in siN3 transfection group were respectively 1.85±0.16, 0.37±0.08, 1.38± 0.11, which were apparently higher than those in negative control group (0.93±0.11, 0.17±0.05, 0.86±0.06; all P<0.05) and blank control group (0.94 ± 0.04, 0.15 ± 0.08, 0.85 ± 0.10, respectively; all P<0.01). CONCLUSIONS: It would be to promote the cell doubling time, reduce cell apoptosis and increase the invasive capability in ES2 cells that ARID1A expression was down-regulating by ARID1A mRNA interference. The invasion mechanism may be related to the activation of NF-κB signal transduction pathway, up-regulation of MT1-MMP expression and then promoting the invasion of tumor cells via the up-regulation of MMP2 expression.

[Clinicopathological features of thyroid papillary microcarcinoma of which the maximum diameter is less than 5 mm: a series of 487 cases].

Objective: To analyze the clinical and pathological features of papillary thyroid microcarcinoma(PTMC) of which the maximum diameter is less than 5 mm. Methods: Clinical data of 487 PTMC (≤5 mm) patients who underwent primary surgery by the same team of doctors between March 2013 to March 2016 at Department of Surgery, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University were analyzed retrospectively. Female patients accounted for 77.4%(377/487). The age ranged from 22 to 80 years, with an average of (46±11) years.χ2 test and multivariate Logistic regression analysis were used to analyze risk factors. Results: There were 93.6% (456/487) of the patients with preoperative definite diagnosis. There were 92.6% (452/487) of the patients underwent unilateral thyroid lobectomy with isthmectomy and ipsilateral central lymph node dissection. Multifocal carcinoma accounted for 20.3% (99/487), Ⅲ to Ⅳ phase cases accounted for 12.3% (60/487), patients with low risk of recurrence accounted for 67.6% (329/487). The central lymph node metastasis rate was 34.9% (170/487). Male (OR=2.149, 95%CI: 1.367 to 3.381), age<45 years (OR=0.451, 95%CI: 0.303 to 0.672), multifocal carcinoma (OR=1.798, 95%CI: 1.116 to 2.898), and capsular invasion (OR=3.678, 95%CI: 1.406 to 9.622) were the independent risk factors for lymph node metastasis (P<0.05). Conclusion: Regardless of tumor size, PTMC still needs to be taken seriously, especially male, age < 45 years, capsular invasion, or multifocal carcinoma patients.

[Comfortable initial periodontal therapy: step by step].

With the transition of the medical model from the traditional biomedical model to the biopsychosocial one, there is a growing trend and requirement for oral operations that prioritize comfort, pain management, minimally invasive techniques, and visualization. Consequently, demands for comfortable dental treatments among individuals are increasing. However, initial periodontal therapy is often accompanied by pain, and patients' reactions to pain range from nervousness to dental fear, such as irritability, hyperventilation, even nausea, vomiting, and refusal to cooperate, which make the implementation of initial periodontal therapy difficult or even impossible. This article will focus on three key steps: firstly, the preparation of the clinic, the acquisition of patients' trust and the implementation of preventive sedation before treatment; secondly, the use of comfort operation and nursing, psychological intervention measures, local anesthesia, and sedation techniques during treatment; thirdly, the health education and follow-up after treatment. By addressing these aspects, we aim to clarify how to perform comfortable initial periodontal therapy step by step.