Mechanism of noncoding RNA-associated N6-methyladenosine recognition by an RNA processing complex during IgH DNA recombination.

Immunoglobulin heavy chain (IgH) locus-associated G-rich long noncoding RNA (SµGLT) is important for physiological and pathological B cell DNA recombination. We demonstrate that the METTL3 enzyme-catalyzed N6-methyladenosine (m6A) RNA modification drives recognition and 3' end processing of SµGLT by the RNA exosome, promoting class switch recombination (CSR) and suppressing chromosomal translocations. The recognition is driven by interaction of the MPP6 adaptor protein with nuclear m6A reader YTHDC1. MPP6 and YTHDC1 promote CSR by recruiting AID and the RNA exosome to actively transcribe SµGLT. Direct suppression of m6A modification of SµGLT or of m6A reader YTHDC1 reduces CSR. Moreover, METTL3, an essential gene for B cell development in the bone marrow and germinal center, suppresses IgH-associated aberrant DNA breaks and prevents genomic instability. Taken together, we propose coordinated and central roles for MPP6, m6A modification, and m6A reader proteins in controlling long noncoding RNA processing, DNA recombination, and development in B cells.

Umbilical double-port laparoscopy combined with extraperitoneal water injection for the treatment of giant inguinal hernias in infants and young children.

OBJECTIVE: Exploration of the efficacy of treating large indirect inguinal hernias in infants and young children using umbilical double-port laparoscopy combined with extraperitoneal water injection. METHODS: A retrospective analysis was conducted on 165 cases of primary unilateral large indirect inguinal hernias in infants and young children treated at our hospital from May 2018 to May 2023. Among them, 90 cases underwent umbilical double-port laparoscopic surgery combined with extraperitoneal water injection and high ligation of the hernia sac (Double-Port Group), and another 75 cases underwent conventional three-port laparoscopic high ligation of the hernia sac (Three-Port Group). The two groups were compared in terms of operation time, postoperative pain scores at 24 hours, hospital stay, incision complications, and recurrence within one year after surgery. RESULTS: Both groups successfully completed the surgery without any intraoperative complications. The pain score at 24 hours postoperatively was lower in the Double-Port Group compared to the Three-Port Group, and there was no statistically significant difference in operation time, hospital stay, and incision complications between the two groups (P > 0.05). Both groups were followed up for one year postoperatively; the Three-Port Group had one recurrence that was cured after further treatment, while there were no recurrences in the Double-Port Group. CONCLUSION: Umbilical double-port laparoscopy combined with extraperitoneal water injection for the treatment of large indirect inguinal hernias in infants and young children has the advantages of being safe and reliable, with concealed and aesthetic incisions, and rapid recovery.

High prevalence of ROS1 gene rearrangement detected by FISH in EGFR and ALK negative lung adenocarcinoma.

ROS1 rearrangement has become an important biomarker for targeted therapy in advanced lung adenocarcinoma (LUAD). The study aimed to evaluate the prevalence of ROS1 rearrangement in Chinese LUAD with EGFR wild-type and ALK fusion-negative status, and analyze the relationship with their clinicopathological characteristics. A large cohort of 589 patients of LUAD with EGFR/ALK wild-type, diagnosed between April 2014 and June 2018, was retrospectively analyzed. ROS1 rearrangement in all these cases was detected by FISH, and 8 selected cases with different positive and negative signals were confirmed by NGS. As a result, total of 56 cases with ROS1 rearrangements out of 589 LUADs (9.51%) were identified by FISH. The frequency of ROS1 rearrangement in women was 22.15% (35/158), which was statistically higher than 4.87% (21/431) in men (P < 0.001). The ROS1 positive rate in the patients with age < 50 years old (25.29%, 22/87) was statistically higher than that in the patients with age ≥ 50 (6.77%, 34/502) (P < 0.001). There was a trend that the frequency of ROS1 rearrangement in LUAD with stage III-IV was higher than that in stage I-II (9.56%, 39/408 vs 2.50%, 1/40), although it did not reach significant difference (P = 0.135). 37 out of 56 cases of ROS1 rearranged LUAD showed solid (n = 20, 35.71%) and invasive mucinous adenocarcinoma (n = 17, 30.36%) pathological subtypes. The median OS for patients of ROS1 rearranged LUAD treated with TKIs (n = 29) was 49.69 months (95% CI: 36.71, 62.67), compared with 32.55 months (95% CI: 23.24, 41.86) for those who did not receive TKI treatment (n = 16) (P = 0.040). The NGS results on ROS1 rearrangement in all the 8 cases were concordant with FISH results. In conclusion, high prevalence of ROS1 rearrangements occurs in EGFR/ALK wild-type LUAD detected by FISH, especially in younger, female, late stage patients, and in histological subtypes of solid and invasive mucinous adenocarcinoma.

Integrative analysis of PD-L1 DNA status, mRNA status and protein status, and their clinicopathological correlation, in diffuse large B-cell lymphoma.

AIMS: The protein expression of programmed death-ligand 1 (PD-L1) has been recognised as being a biomarker for poor prognosis in diffuse large B-cell lymphoma (DLBCL). The aims of this study were to determine PD-L1 DNA status and mRNA status, and to explore whether they contribute to protein expression and their clinicopathological correlation in DLBCL. METHODS AND RESULTS: In this study, we used fluorescence in-situ hybridisation, RNA in-situ hybridisation and immunohistochemistry to determine PD-L1 status at three different levels in 287 DLBCL samples with follow-up. Their correlation and clinical pathological relevance were also analysed. Our results showed that 1.7% (3/175) of patients had PD-L1 DNA amplification, 19.9% (57/287) had high PD-L1 mRNA expression, and 11.8% (34/287) had high PD-L1 protein expression. Both mRNA and protein expression of PD-L1 were significantly higher in non-germinal centre B-cell-like (GCB) DLBCL than in GCB DLBCL (P < 0.05). In addition, the patients with high PD-L1 mRNA or high PD-L1 protein expression but no PD-L1 DNA amplification had significantly poorer overall survival (OS) than those with low PD-L1 expression (P < 0.05). Furthermore, we found that PD-L1 mRNA and PD-L1 protein expression were highly correlated (P = 0.012), which was observed in all three samples with DNA amplification. CONCLUSIONS: PD-L1 DNA amplification is a rare event, PD-L1 mRNA is the main contributor to the high PD-L1 protein expression, and the latter two will serve as important biomarkers for predicting the prognosis of DLBCL patients and selecting them for immunotherapy.

Specific immunohistochemical detection of the BRAF V600E mutation in primary and metastatic papillary thyroid carcinoma.

PURPOSE: To evaluate the clinical value of immunohistochemistry (IHC) using anti-BRAF V600E antibody (clone VE1) for specific detection of the BRAF V600E mutant protein in formalin-fixed paraffin-embedded papillary thyroid carcinoma (PTC) specimens. METHODS: A total of 118 PTC cases and 116 control cases processed between January 2008 and June 2010 were selected and created tissue microarrays (TMAs) for the study. BRAF V600E (VE1) IHC was performed on tissue sections from PTC cases to determine mutation status. Molecular tests (Sanger sequencing/ARMS) were used to confirm the BRAF V600E gene mutation in primary PTC. RESULTS: A uniformly cytoplasmic staining throughout the tumors was observed in IHC-positive cases. BRAF V600E was detected in 68.6% (81/118) of PTC samples by IHC and in 61.9% (73/118) by Sanger sequencing/ARMS. The overall concordance between IHC and Sanger sequencing/ARMS was 93.2% (110/118). The sensitivity and specificity of the BRAF V600E IHC was 100% (73/73) and 82.2% (37/45), respectively. Positive and negative predictive values were 90.1% (73/81) and 100% (37/37), respectively. Expression of the BRAF V600E mutant protein was detected in all of 59 cases of primary carcinoma and corresponding metastatic carcinoma in lymph nodes. The concordance between IHC staining in primary and metastatic PTC was 100% (59/59). CONCLUSION: IHC using VE1 antibody for detection of the BRAF V600E mutant protein expression in PTC showed high sensitivity and acceptable specificity, which are critical for diagnostic purposes. IHC staining for BRAF V600E showed uniformly cytoplasmic expression in both primary tumor and metastatic nodes. Therefore, IHC has high practical value for the detection of the BRAF V600E mutation in metastatic and primary PTC.

Microcystic stromal tumour of the ovary: frequent mutations of ß-catenin (CTNNB1) in six cases.

AIMS: To analyse the clinicopathological, immunohistochemical and ß-catenin (CTNNB1) mutation characteristics of six cases of microcystic stromal tumour of the ovary (MCST). METHODS AND RESULTS: Six Chinese patients with MCST who ranged in age from 29 to 69 years (mean 50 years) were included in the study. Five patients were detected with a pelvic mass during routine health examinations and one patient presented initially with opsomenorrhoea. All tumours involved the left ovary, with solid-cystic cut surface in five cases and cystic cut surface in one case. Microscopically, microcysts, solid nests and hyaline degenerated fibrous stroma were variably mixed. Immunohistochemically, the tumour cells in all cases were diffusely positive for CD10, vimentin and WT-1 and negative for α-inhibin and calretinin. ß-catenin expression was observed in both the nucleus and the cytoplasm in five cases and only in the cytoplasm in one case. The results of CTNNB1 mutation analysis revealed four missense point mutations in four cases, which were c.97T>C, c.101G>A, c.110C>G and c.122C>T. CONCLUSIONS: MCST shows a unique morphology with characteristic immunophenotype. ß-catenin expression in the nucleus and ß-catenin mutations were identified in the majority of cases, which suggests that the Wnt/ß-catenin pathway may play a crucial role in the tumorigenesis of MCST.

[Study on the relations among the screen-based sedentary behaviors, family factors and body mass index of children].

OBJECTIVE: To explore the relations among screen-based sedentary behaviors (SSB), family factors and body mass index (BMI) of children, and to study how family factors have effect on BMI through influencing SSB. METHODS: A total of 1,846 students aged 7-11 years from 12 primary schools in one district of Beijing were included. Their body weight and height were measured to calculate the BMI. The time of SSB and family factors were investigated by using questionnaires. The time of SSB was the total time of watching TV and videos, playing computer games and iPad each day during the past 7 days recalled by children. The family factors included the parents' education, occupation, the parents'time of SSB, whether the parents told their child the harm of SSB, the parents'time limit for the children's SSB. The parents'education and occupation were used for calculating the family socioeconomic score. RESULTS: The median time of SSB for children was 1 hour/day, and the interquartile range was 1 hour/day. The BMI of the children with the parents' time limit for the children's SSB less than 120 min/day were smaller than the children with the parents'time limit not less than 120 min/day, in both the boys (1.63 kg/m2, P<0.001) and the girls (0.85 kg/m2, P=0.004). The family socioeconomic score, the parents'SSB time, whether the parents told their children the harm of SSB were not related to the children's BMI . The mediation effects of SSB time for children on the association between the parents'time limit for the children's SSB and BMI were -0.222 kg/m2 (95%CI:-0.432, -0.095) for boys and -0.187 kg/m2 (95%CI: -0.507, -0.049) for girls, which accounted for 13.67% of the total effects for boys and 22.11% for girls. CONCLUSION: The parents' time limit for the children's SSB has effect on their BMI through influencing their SSB time. Parents' supervision on the behaviors of children produces larger benefit for BMI than health education conveyed by parents. Therefore, parents' participation in supervising the behaviors of children are indispensable for preventing and controlling childhood obesity.

Effectiveness of a school-based physical activity intervention on obesity in school children: a nonrandomized controlled trial.

BACKGROUND: Childhood obesity has been a serious public health problem. An effective school-based physical activity (PA) intervention is still lacking in China. This study aimed to assess the effectiveness of a school-based physical activity intervention during 12 weeks on obesity and related health outcomes in school children. METHODS: It was a non-randomized controlled trial. Altogether 921 children aged 7 to 15 years were recruited at baseline survey. Children in the intervention group (n = 388) participated in a multi-component physical activity intervention during 12 weeks that included improvement of physical education, extracurricular physical activities for overweight/obese students, physical activities at home, and health education lectures for students and parents. Children (n = 533) in the control group participated in usual practice. RESULTS: Participants had mean age of 10.4 years, mean body mass index (BMI) of 19.59 kg/m2, and 36.8 % of them were overweight or obese at baseline survey. The change in BMI in intervention group (-0.02 ± 0.06 kg/m2) was significantly different from that in control group (0.41 ± 0.08 kg/m2). The adjusted mean difference was -0.43 kg/m2 (95% CI: -0.63 to -0.23 kg/m2, P < 0.001). The effects on triceps, subscapular, abdominal skinfold thickness and fasting glucose were also significant in intervention group compared with control group (all P < 0.05). The change in duration of moderate to vigorous physical activity (MVPA) in intervention group (8.9 ± 4.3 min/day) was significantly different from that in control group (-13.8 ± 3.3 min/day). The adjusted mean difference was 22.7 min/day (95% CI: 12.2 to 33.2 min/day, P < 0.001). CONCLUSIONS: The school-based, multi-component physical activity intervention was effective to decreasing levels of BMI, skinfold thickness, fasting glucose and increasing duration of MVPA. These findings provided evidence for the development of effective and feasible school-based obesity interventions. TRIAL REGISTRATION: Clinicaltrials.gov Identifier: NCT02074332 (2014-02-26).

Association between physical activity and the prevalence of gout among patients with type 2 diabetes mellitus and hyperuricemia: a two-center population-based cross-sectional study.

INTRODUCTION: Diabetes mellitus (DM) and gout cohabitation severely reduces patient life quality while raising financial burden on individual and society. The aim of this study was to elucidate the association between physical activity (PA) and the prevalence of gout among type 2 DM (T2DM) and hyperuricemia (HUA) patients. METHODS: In all, we recruited 2291 T2DM patients with HUA. Among them, 448 had gout and 1843 did not. We collected patient data, such as anthropometry, laboratory reports, and medical history, for our analyses. The PA assessment was based on the Chinese version of International PA Questionnaire-short (IPAQ). Moreover, the relationship between PA and gout risk was examined using multivariate logistic regression models. RESULTS: Total PA was markedly low among gout patients, relative to controls (p < 0.05). Based on the IPAQ categorical score, 38.2% exhibited "low," 26.8% "moderate," and 35.0% "high" PA among gout patients. In comparison, 12.4% performed "low," 53.8% "moderate," and 33.8% "high" PA among controls. Multivariate analysis revealed that, after adjustment of confounding factors, both low (OR 6.382) and high PA (OR 2.048) had a higher prevalence of gout, as compared to moderate PA. Moreover, we revealed that the male sex, age, HUA duration, serum uric acid, glycated hemoglobin, dyslipidemia history, and drinking status were also independent indicators of the prevalence of gout. Furthermore, stratification analyses revealed results consistent with our prior results. CONCLUSIONS: PA intensity was associated with the prevalence of gout among T2DM and HUA patients, and the lowest prevalence was achieved from moderate intensity PA. Key Points • PA intensity was associated with the prevalence of gout among T2DM and HUA patients. • The lowest prevalence of gout was achieved from moderate intensity PA.

[Association between abdominal obesity and blood pressure among 7 to 10 years old Chinese children].

OBJECTIVE: To analyze the association between abdominal obesity and blood pressure among 7-10 years old Chinese children. METHODS: A total of 40 495 children aged 7-10 years with complete height, weight, waist circumference and blood pressure data were chosen from the data of 2010 Chinese National Survey on Students Constitution and Health. Based on the "Reference Norms for Screening Overweight and Obesity in Chinese Children and Adolescents" developed by Working Group Obesity in China (WGOC) and the waist to height ratio reference value for children (WHtR ≤ 0.46) , the children were divided into 4 body types (normal weight, simple abdominal obesity, combined obesity and other types). High blood pressure was defined as the systolic blood pressure (SBP) and/or diastolic blood pressure (DBP) higher than the P 95 blood pressure value of 2010 national student of the same gender and age. The prevalence of simple abdominal obesity, combined obesity and high blood pressure were described. Three groups (with 2165 children in each) of children with normal weight, simple abdominal obesity, combined obesity respectively were selected from the chosen ones matched by gender, age (within ± 0.5 years old) and height (within ± 0.3 cm) at the ratio of 1: 1: 1. The value of blood pressure and prevalence of high blood pressure were described.χ(2) test, analysis of variance and multilevel models were used to analyze the relationship between obesity and blood pressure. RESULTS: A total of 40 475 children were selected from the database, including 20 175 boys and 20 320 girls. The prevalence of simple abdominal obesity was 6.36% (2576/40 495), the prevalence of boys and girls was 7.41% (1494/20 175) and 5.32% (1082/20 320) respectively. The prevalence of combined obesity was 16.33% (6611/40 495), the prevalence of boys and girls was 21.30% (4298/20 175) and 11.38% (2313/20 320) respectively. There were statistical differences in the body type distribution of both boys and girls (χ(2) = 869.01, P < 0.01). The prevalence of high blood pressure was 9.62% (3896/40 495), the prevalence of boys (10.05% (2028/20 175)) was higher than girls (9.19% (1868/20 320)) (χ(2) = 8.59, P < 0.01). The value of SBP and DBP among combined obesity group (boys: (103.8 ± 11.3), (64.7 ± 10.1) mm Hg (1 mm Hg = 0.133 kPa); girls: (102.9 ± 12.1), (64.5 ± 10.0) mm Hg) > simple abdominal obesity group (boys: (99.5 ± 11.2), (61.6 ± 9.9) mm Hg; girls: (99.6 ± 11.4), (62.6 ± 9.3) mm Hg)> normal weight group (boys: (97.4 ± 10.8), (60.5 ± 9.4) mm Hg; girls: (97.2 ± 10.8), (60.8 ± 9.4) mm Hg), and the differences were statistical significant (Fboys: 113.22, 62.05; Fgirls: 54.19, 32.31, P < 0.01). According to the results of multilevel model, among boys, compared with the normal weight group, the SBP of combined obesity group and simple abdominal obesity group was 6.0 and 1.8 mm Hg higher respectively (Wald χ(2): 17.55, 204.94, P < 0.01); the DBP was 4.0 and 0.9 mm Hg higher respectively (Wald χ(2): 6.37, 114.05, P < 0.05). Among girls, the SBP was 5.0 and 2.1 mm Hg higher respectively (Wald χ(2):16.47, 92.52, P < 0.01); the DBP was 3.5 and 1.6 mm Hg higher respectively (Wald χ(2): 12.29, 57.52, P < 0.01). Comparing with normal group, the risk of high SBP among boys with simple abdominal obesity was higher (OR = 1.48; 95%CI: 1.06-2.06), and both the boys (SBP: OR = 3.06; 95%CI: 2.28-4.11) ; DBP: OR = 2.72; 95%CI: 1.99-3.72) and girls (SBP: OR = 2.48; 95%CI: 1.75-3.53; DBP: OR = 2.64; 95%CI: 1.82-3.93) in combined obesity group had a higher risk of high SBP and high DBP. CONCLUSION: Simple abdominal obesity is associated with the increasing of blood pressure in children, and combined obesity has a closer ties than simple abdominal obesity.

[Analysis of influencing factors on physical endurance of Chinese primary and middle school students aged 10 to 17].

OBJECTIVE: To explore the influencing factors on physical endurance among Chinese primary and middle school students aged 10-17. METHODS: Data was used from "2010 National Physical Fitness and Health Surveillance", from which 68 458 primary and middle school students with complete information were selected. Good physical endurance was defined as endurance running time ≤ P 25 of same gender and age, poor physical endurance as ≥ P 75 of same gender and age. Difference of endurance running time in different physical endurance groups was compared; chi-square test, logistic regression and multiple level model were used to analyze influence of region, BMI, diet, exercise and exercise willingness on physical endurance. RESULTS: There were 34 293 (50.0%) students of good physical endurance and 34 219 (50.0%) ones of poor physical endurance in 68 458 subjects. Percentage of good physical endurance was 53.8% (18 285/33 996) in rural students, higher than urban ones (46.3% (15 954/34 462)) (χ(2) = 384.16, P < 0.01). Percentage of good physical endurance was 12.2% (448/3660) in obese students, lower than normal (55.2% (28 183/51 028)), overweight (29.3% (2012/6874)) and emaciated ones (52.1% (3596/6896)) (χ(2) = 3840.08, P < 0.01). Percentage of good physical endurance was 52.5% (25 534/48 679) in students having breakfast everyday, higher than ones never having breakfast or 1-2 times per week (41.3% (3209/7768))(χ(2) = 438.31, P < 0.01). Percentage of good physical endurance was 51.7% (11 580/22 379) in students having milk everyday, slightly higher than ones never or sometimes having milk (49.2% (22 659/46 079)) (χ(2) = 39.84, P < 0.01). Percentage of good physical endurance was 55.3% (18 961/34 314) in students with adequate physical education class, higher than ones with inadequate class (44.7% (15 278/34 144)) (χ(2) = 756.41, P < 0.01). Percentage of good physical endurance was 58.6% (9484/16 195) in students with one hour exercise or more per day, higher than ones with less than one hour exercise (47.4% (24 755/52 263)) (χ(2) = 619.82, P < 0.01). Percentage of good physical endurance was 60.1% (19 287/32 118) in students who were "very willing" to take part in endurance running, higher than "generally willing" ones (45.4% (10 569/23 272)) and "not willing" ones (33.5% (4383/13 068)) (χ(2) = 2909.61, P < 0.01). Multiple level analysis revealed that risk of poor physical endurance was lower in rural students (OR = 0.80, 95%CI: 0.77-0.82) than urban ones. Risk in emaciated students (OR = 1.11, 95%CI: 1.05-1.17), overweight students (OR = 2.69, 95%CI: 2.55-2.85) and obese students (OR = 7.71, 95%CI: 6.97-8.52) was higher than normal ones. Concerning diet, risk of poor physical endurance in students never having breakfast or once to twice per week was highest (OR = 1.20, 95%CI: 1.13-1.26), followed by students having breakfast three to five times per week (OR = 1.13, 95%CI: 1.08-1.18); risk in students having one bag of milk per day was higher (OR = 1.09, 95%CI: 1.02-1.17) than ones having two bags. Concerning exercise, risk of poor physical endurance in students with inadequate physical education class (OR = 1.20, 95%CI: 1.13-1.26) and less than one hour exercise per day (OR = 1.34, 95%CI: 1.29-1.40) was higher than ones with adequate physical education class and one hour exercise or more respectively. Risk of poor physical endurance in students who were "generally willing" (OR = 1.67, 95%CI: 1.61-1.73) and "not willing" (OR = 2.72, 95%CI: 2.60-2.85) to take part in endurance running was higher than "very willing" ones. CONCLUSION: Urban and rural residence, BMI, breakfast, milk, physical education class, exercise time and exercise willingness were related to primary and middle school students'physical endurance.

[Association between early menarche and physical fitness among adolescent girls in China].

OBJECTIVE: To explore the association between early menarche and physical fitness among adolescent girls in China. METHODS: Research material was selected from the data of "2010 National Physical Fitness and Health Survey". Probability unit regression method was used to calculate the age of 10th percentile at menarche and menarche age before the 10th percentile was defined as early menarche(9.0-11.6 years old). A total of 1072 girls with early menarche were selected. Each girl with early menarche was matched with one girl from the same urban or rural locations who hadn't achieved menarche with the age difference less than 0.1 years. A total of 1072 girls without menarche were selected. Indicators of physical fitness included 50 m running, standing broad jump, 50 m×8 shuttle running and sit-ups. Differences of physical fitness between early menarche and without menarche were analyzed using t test stratified by age and urban/rural area. The multilevel models of single dependent variable and multiple dependent variables were used to analyze association between early menarche and physical fitness. RESULTS: A total of 2144 students were put in this research. Among girls (11.0-11.6 years old) in urban areas, 50 m running, standing broad jump and sit-ups was (9.78 ± 0.85) s, (153.81 ± 18.59) cm, and 27.79 ± 10.25, respectively for those with early menarche, while in girls without menarche was (10.01 ± 0.90) s, (149.71 ± 18.72) cm and 26.28 ± 10.11, respectively. There were significant differences between two groups on all above variables (t values were 4.02, 3.43 and 2.31, respectively with all P values <0.01). Among girls in rural, 50 m×8 shuttle-running and sit-ups was (125.22 ± 15.57) s and 24.96 ± 8.97 for those with early menarche, while it was (120.92 ± 13.06) s and 22.96 ± 9.83 for those without menarche. There were significant differences between two groups on both variables(t values were 3.89 and 2.77 with both P values < 0.01). In addition, 50 m×8 shuttle-running in girls (10.0-10.9 years old) with early menarche was (128.52 ± 15.74) s and it was (123.89 ± 13.50) s in girls without menarche. The difference was significant (t = 2.14, P < 0.05). The multilevel analysis showed that 50 m running, standing broad jump and sit-ups in girls with early menarche was 0.12 s, 3.14 cm and 1.11 higher than girls without menarche (Waldχ(2) values were 4.00, 6.22 and 4.07, respectively with all P values < 0.05). But 50 m×8 shuttle-running in girls with early menarche was 1.95 s less than girls without menarche (Waldχ(2) = 3.96, P < 0.05). CONCLUSION: Early menarche may be associated with higher speed fitness, leg power and muscle power and lower physical stamina.

Noncoding mutations cause super-enhancer retargeting resulting in protein synthesis dysregulation during B cell lymphoma progression.

Whole-genome sequencing of longitudinal tumor pairs representing transformation of follicular lymphoma to high-grade B cell lymphoma with MYC and BCL2 rearrangements (double-hit lymphoma) identified coding and noncoding genomic alterations acquired during lymphoma progression. Many of these transformation-associated alterations recurrently and focally occur at topologically associating domain resident regulatory DNA elements, including H3K4me3 promoter marks located within H3K27ac super-enhancer clusters in B cell non-Hodgkin lymphoma. One region found to undergo recurrent alteration upon transformation overlaps a super-enhancer affecting the expression of the PAX5/ZCCHC7 gene pair. ZCCHC7 encodes a subunit of the Trf4/5-Air1/2-Mtr4 polyadenylation-like complex and demonstrated copy number gain, chromosomal translocation and enhancer retargeting-mediated transcriptional upregulation upon lymphoma transformation. Consequently, lymphoma cells demonstrate nucleolar dysregulation via altered noncoding 5.8S ribosomal RNA processing. We find that a noncoding mutation acquired during lymphoma progression affects noncoding rRNA processing, thereby rewiring protein synthesis leading to oncogenic changes in the lymphoma proteome.

Analysis of the Effectiveness of Nursing Interventions in Critically Ill Patients in Respiratory Medicine.

OBJECTIVE: To enhance and analyze the clinical effectiveness of implementing quality nursing interventions in the clinical care of critically ill patients in respiratory medicine. METHODS: Clinical data of respiratory medicine patients treated in our hospital over the years were collected and 96 patients who met the requirements of the purpose of this study and the sample inclusion criteria were selected as the study subjects from the patients treated between April 2019 and January 2020. According to the care methods received by the patients in our hospital, 48 of them who implemented conventional care were used as the control group, and another 48 patients who were given quality care interventions were used as the observation group. The data were statistically recorded and comparatively analyzed for the indicators such as nursing oxygen index, heart rate, and clinical treatment efficiency of patients in both groups. RESULTS: Compared with the control group, patients in the observation group who received quality nursing intervention had more significant improvement in blood oxygen index and heart rate after nursing care; the clinical treatment efficiency of patients in the observation group was significantly higher than that of the control group (95.83% vs. 81.25%). The data comparison between the groups showed a significant difference, P < 0.05, which was statistically significant. CONCLUSION: Adding quality nursing interventions to the implementation of conventional care for patients with respiratory diseases can better improve patients' clinical symptoms, accelerate their clinical recovery, improve and enhance prognosis, and further improve clinical outcomes.

High glucose inhibits the osteogenic differentiation of periodontal ligament stem cells in periodontitis by activating endoplasmic reticulum stress.

Background: Periodontitis is a highly prevalent dental disease which is associated with diabetes and is challenging to cure in diabetic patients. However, the mechanism of comorbid diabetes and periodontitis is still unclear. This study aimed to uncover the role of endoplasmic reticulum (ER) stress in high glucose-associated periodontitis. Methods: Periodontal tissues were obtained from diabetic patients with periodontitis, periodontitis patients without systemic disease, and healthy teeth. The expressions of ER stress-related factors GRP78, ATF6, PERK and XBP1 were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and immumohistochemical staining. Periodontal ligament stem cells (PDLSCs) from three states of periodontal tissues were isolated and cultured as diabetic PDLSCs (dPDLSCs), inflamed PDLSCs (iPDLSCs) and healthy PDLSCs (hPDLSCs), and the cell stemness was assayed. Different concentrations (8, 11, and 25 mmol/L) of D-glucose were used on hPDLSCs to simulate high glucose microenvironment. The changes of osteogenic ability of PDLSCs were observed, and the expressions of ER stress-related factors in different time point (6, 12, 24, and 72 h) were detected. Finally, GRP78 shRNA lentivirus was used to block ER stress on PDLSCs in the 25 mmol/L D-glucose microenvironment, and the osteogenic ability of PDLSCs was observed. Results: The results showed that the expressions of GRP78, ATF6, PERK, and XBP1 were highest in the diabetic periodontitis group and lowest in the healthy periodontal tissue group (P<0.05). The clone formation, osteogenic and lipogenic differentiation abilities were lowest in dPDLSCs and highest in hPDLSCs. With the increase of glucose concentration, the osteogensis ability of PDLSCs decreased. After 6 hours of stimulation with D-glucose 25 mmol/L, the ER stress pathways in PDLSCs were effectively activated, and the peak value was reached at 12 hours. The decrease in the osteogensis ability of PDLSCs in a high glucose microenvironment reversed when ER stress was blocked. Conclusions: The osteogenic differentiation ability of PDLSCs cells is inhibited in a high glucose microenvironment, and this effect is realized by ER stress activation. Blocking ER stress can partially restore the reduced osteogenic ability of PDLSCs. These results suggest that high glucose inhibits the osteogenic differentiation ability of PDLSCs by activating ER stress, which ultimately exacerbates periodontitis.

RNA exosome drives early B cell development via noncoding RNA processing mechanisms.

B cell development is linked to successful V(D)J recombination, allowing B cell receptor expression and ultimately antibody secretion for adaptive immunity. Germline noncoding RNAs (ncRNAs) are produced at immunoglobulin (Ig) loci during V(D)J recombination, but their function and posttranscriptional regulation are incompletely understood. Patients with trichohepatoenteric syndrome, characterized by RNA exosome pathway component mutations, exhibit lymphopenia, thus demonstrating the importance of ncRNA surveillance in B cell development in humans. To understand the role of RNA exosome in early B cell development in greater detail, we generated mouse models harboring a B cell-specific cre allele (Mb1cre), coupled to conditional inversion-deletion alleles of one RNA exosome core component (Exosc3) or RNase catalytic subunits (Exosc10 or Dis3). We noticed increased expression of RNA exosome subunits during V(D)J recombination, whereas a B cell developmental blockade at the pro-B cell stage was observed in the different knockout mice, overlapping with a lack of productive rearrangements of VDJ genes at the Ig heavy chain (Igh). This unsuccessful recombination prevented differentiation into pre-B cells, with accumulation of ncRNAs and up-regulation of the p53 pathway. Introduction of a prearranged Igh VDJ allele partly rescued the pre-B cell population in Dis3-deficient cells, although V-J recombination defects were observed at Ig light chain kappa (Igκ), preventing subsequent B cell development. These observations demonstrated that the RNA exosome complex is important for Igh and Igκ recombination and establish the relevance of RNA processing for optimal diversification at these loci during B cell development.

Noncoding RNA processing by DIS3 regulates chromosomal architecture and somatic hypermutation in B cells.

Noncoding RNAs are exquisitely titrated by the cellular RNA surveillance machinery for regulating diverse biological processes. The RNA exosome, the predominant 3' RNA exoribonuclease in mammalian cells, is composed of nine core and two catalytic subunits. Here, we developed a mouse model with a conditional allele to study the RNA exosome catalytic subunit DIS3. In DIS3-deficient B cells, integrity of the immunoglobulin heavy chain (Igh) locus in its topologically associating domain is affected, with accumulation of DNA-associated RNAs flanking CTCF-binding elements, decreased CTCF binding to CTCF-binding elements and disorganized cohesin localization. DIS3-deficient B cells also accumulate activation-induced cytidine deaminase-mediated asymmetric nicks, altering somatic hypermutation patterns and increasing microhomology-mediated end-joining DNA repair. Altered mutation patterns and Igh architectural defects in DIS3-deficient B cells lead to decreased class-switch recombination but increased chromosomal translocations. Our observations of DIS3-mediated architectural regulation at the Igh locus are reflected genome wide, thus providing evidence that noncoding RNA processing is an important mechanism for controlling genome organization.

Surveillance of Borrelia miyamotoi-carrying ticks and genomic analysis of isolates in Inner Mongolia, China.

BACKGROUND: Borrelia miyamotoi is a newly described relapsing fever spirochete transmitted by ixodid tick species. Little is known about the prevalence of B. miyamotoi infections in humans and ticks in Inner Mongolia, China. Therefore, we investigated the prevalence of B. miyamotoi in Ixodes persulcatus ticks, and we aimed to isolateB. miyamotoi from I. persulcatus from four regions of Greater Khingan, Inner Mongolia, China. METHODS: From May to June each year during the period 2016-2019, host-seeking adult I. persulcatus ticks were collected from vegetation. Genomic DNA was prepared from half of each tick body for PCR template, and the remaining half was used to cultivate B. miyamotoi in BSK-M medium. We employed quantitative real-time PCR (qPCR) to detect Borrelia DNA in the ticks and to calculate the prevalence of B. miyamotoi and infections with other borreliae. For characterization of the isolated B. miyamotoi, we performed draft genome sequencing and multilocus sequencing analysis (MLSA). RESULTS: A total of 2656 adult I. persulcatus ticks were collected. The overall prevalence of relapsing fever (RF) borreliae in ticks was 5.0% (134/2656) and that of Lyme disease (LD) borreliae was 43.8% (1164/2656). Co-infection with RF and LD borreliae was observed in 63 ticks (2.4%). Ticks that were positive for RF borreliae by qPCR were subjected to glycerophosphodiester diester phosphodiesterase gene (glpQ) PCR amplification and sequencing, through which we identified the RF borrelia specimens as B. miyamotoi. Furthermore, the B. miyamotoi strain Hetao-1 was isolated from I. persulcatus, and a draft genome sequence was obtained from the isolate. Sequencing determined the strain Hetao-1 genome to be approximately 906.1 kbp in length (28.9% average GC content), and MLSA identified the strain as ST633, which has previously been reported in Japan and Mongolia. CONCLUSION: We detected B. miyamotoi from I. persulcatus ticks collected in Inner Mongolia, and successfully isolated a B. miyamotoi strain. To our knowledge, this is the first study to culture a B. miyamotoi isolate from China. The data on the prevalence of B. miyamotoi and other borreliae in I. persulcatus ticks will be fundamental for future epidemiological studies of B. miyamotoi disease in Inner Mongolia.

Correlation between clinicopathological features and KRAS, NRAS, and BRAF mutation status in Chinese colorectal cancer patients.

This study was retrospectively performed to analyze correlations between clinicopathological features of colorectal cancer (CRC) and mutations in KRAS, NRAS, and BRAF in Chinese patients, and to assess the importance of detecting additional mutations in KRAS exons 3 and 4 and NRAS in patients with CRC. RAS (KRAS and NRAS) and BRAF mutations were detected in 715 and 655 patients respectively. The mutation rate of RAS (KRAS or NRAS) was 45.6% (326/715). KRAS exon 2 mutations were evaluated in 36.6% of patients (262/715). Additional mutations in RAS exons occurred in 9.0% of patients (64/715), including KRAS exons 3 and 4 in 5.6% (40/715) and NRAS exons 2, 3, or 4 in 3.4% (24/715). Among 453 patients with wild-type KRAS exon 2, 14.1% (64/453) had other mutations in RAS exons. The most frequent sites of mutations were codons 12, 13, 61, and 146 in KRAS and codons 12 and 61 in NRAS. The mutation rate of BRAF (exon 15) was 4.0% (26/655), and the most frequent mutation site was codon 600. Among 440 patients with CRC who had a primary tumor resection at our center, those with mucinous or signet ring cell CRC were more likely to harbor KRAS mutations than those with adenocarcinoma (62.7% vs. 43.6%, P=0.006 and 59.3% vs. 39.6%, P=0.004, respectively). Female patients had a higher BRAF (exon 15) mutation rate than male patients (5.1% vs. 1.1%, P=0.017). Detection of both KRAS and NRAS mutations is useful for selecting patients who will benefit from anti-EGFR monoclonal antibody therapy. KRAS mutations were more frequent in patients with mucinous adenocarcinoma/signet ring cell CRC, whereas BRAF mutations were more common in female patients with CRC.

Expression and function of CXCL12/CXCR4/CXCR7 in thyroid cancer.

The contribution of CXCL12/CXCR4/CXCR7 axis to cancer progression has been increasingly recognized. However, its role in thyroid cancer development remains unclear. The present study aimed to examine the expression and function of CXCL12 and its receptors in thyroid cancer. The expression of CXCL12/CXCR4/CXCR7 in human tissue specimens of papillary, follicular, medullary, and anaplastic thyroid carcinoma, follicular adenoma, Hashimoto's thyroiditis and nodular goiter were examined by immunohistochemistry using a tissue microarray. CXCR4 and CXCR7 were over-expressed in human thyroid cancer cells K1 by transduction of recombinant lentivirus. The effect of overexpression of CXCR4 and CXCR7 on K1 cell proliferation and invasion and the molecular mechanism underlying the effect were investigated. CXCL12 was exclusively expressed in papillary thyroid carcinoma tissue but absent in other types of thyroid malignancies and benign lesions. CXCR7 was widely expressed in the endothelial cells of all types of malignancy but only occasionally detected in benign lesions. CXCR4 was expressed in 62.5% of papillary thyroid carcinoma tissue specimens and in 30-40% of other types of malignancy, and it was either absent or weakly expressed in benign lesions. CXCL12 stimulated the invasion and migration of K1 cells overexpressing CXCR4, but did not affect K1 cells overexpressing CXCR7. K1 cell proliferation was not affected by overexpression of CXCR4 or CXCR7. Overexpression of CXCR4 in K1 cells significantly increased AKT and ERK phosphorylation and markedly induced the expression and activity of matrix metalloproteinase-2 (MMP­2). Thus, CXCL12 may be an effective diagnostic marker for papillary thyroid carcinoma, and CXCL12/CXCR4/CXCR7 axis may contribute to thyroid cancer development by regulating cancer cell migration and invasion via AKT and ERK signaling and MMP-2 activation.