[Bioinformatics analysis in metagenomic next-generation sequencing of pathogenic microorganisms: current status and challenges].

The clinical application of metagenomic next-generation sequencing (mNGS) in the diagnosis of unknown pathogenic infections and critical infections has become increasingly valuable. Due to the huge volume of mNGS data and the complexity of clinical diagnosis and treatment, mNGS has difficulties in data analysis and interpretation in practical application. Therefore, in the process of clinical practice, it is crucial to grasp the key points of bioinformatics analysis and establish a standardized bioinformatics analysis process, which is an important step in the transformation of mNGS from laboratory to clinic. At present, bioinformatics analysis of mNGS has made great progress, but with the high requirements of clinical standardization of bioinformatics analysis and the development of computer technology, bioinformatics analysis of mNGS is also facing new challenges. This article mainly elaborates on quality control, and identification and visualization of pathogenic bacteria.

[Efficacy of video-assisted thoracoscopic surgical decortication for stage â ¢ tuberculous empyema].

Objective: To investigate the clinical efficacy, safety and feasibility of "double-portal" video-assisted thoracoscopic surgical(VATS) decortication among patients with stage Ⅲ tuberculous empyema, and then to evaluate the recovery of chest deformity. Method: This study was a single center retrospective study. A total of 49 patients with stage Ⅲ tuberculous empyema who underwent VATS pleural decortication at the Department of Thoracic Surgery, Public Health Clinical Center of Chengdu between June 2017 and April 2021 were enrolled, including 38 males, and 11 females, aged 13-60 (27.5±10.4) years. The safety and feasibility of VATS were further evaluated. The inner circumference of the chest on sternal and xiphoid planes on chest CT scans before and 1, 3, 6, 12months after decortication were collected through the measuring software of the CT. The samples in-pair test was used to compare the changes in the chest to reflect the recovery of the chest deformity. Results: In the 49 patients, The surgical time was (186±61) min, and the volume of blood loss was (366±267) ml. There were 8 cases (16.33%) with postoperative complications during the perioperative period. Constant air leak and pneumonia were the main postoperative complications. No relapse of empyema or dissemination of tuberculosis occured during the period of follow-up. Before surgery, the inner thoracic circumference of the thorax at the level of the carina plane was (655±54) mm, and the inner thoracic circumference of the thorax at the level of the xiphoid plane was (720±69) mm. Patients were followed for 12-36 months. The inner thoracic circumference of the thoracic cavity at the level of carina was (666±51), (667±47) and (671±47) mm at the 3rd, 6th and 12th months after operation, which were significantly larger than that at the level of carina before operation (all P<0.05). The inner thoracic circumference diameter of the thoracic cavity measured at the xiphoid level at the 3rd, 6th and 12th months after the operation was (730±65), (733±63) and (735±63) mm respectively(all P<0.05).The inner thoracic circumference of the thoracic cavity increased significantly than that before surgery (P<0.05). At 6 months after operation, there was significant difference in the improvement of the inner thoracic circumference of the carina plane in patients with age less than 20 years and FEV1% less than 80% (P=0.015, P=0.003). The improvement in the inner thoracic circumference of the carina plane in patients with pleural thickening≥8 mm compared with those with less than 8 mm was not statistically different(P=0.070). Conclusions: For some patients with stage Ⅲ tuberculous empyema, pleural decortication under thoracoscopy is safe and feasible, and can significantly restore the inner thoracic circumference of the patient's chest, improve the collapse of the patient's chest, and have significant clinical effect. The "double-portal VATS" surgical technology has the advantage of less trauma, wide operation field, large operation space and is easy to master, which is worth further exploring for clinical application.

[International multi-center evaluation of a rapid antigen test based on gold immunochromatographic assay for detection of severe acute respiratory syndrome coronavirus 2].

Objective: The gold immunochromatographic assay for detection of SARS-CoV-2 antigen was evaluated by international multi-center clinical trial. Methods: A total of 1 855 clinical parallel samples with valid test results (for nucleic acid and antigen tests, respectively) were collected from nine countries, including Germany, the United Kingdom, Ukraine, France, India, Thailand, Malaysia, the United States of America and Brazil, with sampling period from January 3, 2021 to September 22, 2021. These samples were detected by SARS-CoV-2 antigen test kit (colloidal gold immunochromatography assay) and nucleic acid detection kit (real-time fluorescent quantitative reverse transcription polymerase chain reaction). Positive coincidence rates [(number of antigen-positive cases/nucleic acid-positive cases)×100%], negative coincidence rates [(number of antigen-negative cases/nucleic acid-negative cases)×100%], total coincidence rates [(number of cases with consistent results for both antigen and nucleic acid detection/number of total cases) ×100%], as well as Kappa values were calculated. The differences of the above indictors among different countries were evaluated by the coefficient of variation. The detection rates of the antigen test for samples with different cycle threshold values (Ct values) for the nucleic acid detection, different characteristics and different mutant strains were analyzed. Results: For all samples, the positive, negative, and total coincidence rate between the antigen test and nucleic acid assay was 90.8% (569/627), 99.7% (1 224/1 228) and 96.7% (1 793/1 855), respectively, and the consistency coefficient Kappa value was 0.924. Among these countries, the coefficient of variation for positive coincidence rates (except for Malaysia with a lot of samples with Ct value>30), negative coincidence rates (except for France without negative samples) and total coincidence rates (except for France) was 6%,<1%, and 6%, respectively. When Ct values were less than 25, the detection rates of antigen test were 83.3%-100% for each countries (the coefficient of variation was 6%); The total detection rate and the coefficient of variation was 93.4% (428/458) and 5%, respectively, for asymptomatic infected persons and cases within 7 days post onset of symptoms; the total detection rate for various SARS-CoV-2 mutant strains was 97.5% (119/122); and it showed negative results for samples from cases infected with other viruses, including influenza A virus subtype H1N1, influenza B virus, respiratory syncytial virus subgroups A and B, coxsackievirus 16, human metapneumovirus, parainfluenza virus types 1 and 4, Epstein-Barr virus and adenovirus. Conclusion: The SARS-CoV-2 antigen test kit showed excellent authenticity, and there were few differences for its indictors among nine countries, therefore it can meet the needs of large-scale early screening of SARS-CoV-2 infection.

[International multi-center evaluation of a rapid antigen test based on gold immunochromatographic assay for detection of severe acute respiratory syndrome coronavirus 2].

Objective: The gold immunochromatographic assay for detection of SARS-CoV-2 antigen was evaluated by international multi-center clinical trial. Methods: A total of 1 855 clinical parallel samples with valid test results (for nucleic acid and antigen tests, respectively) were collected from nine countries, including Germany, the United Kingdom, Ukraine, France, India, Thailand, Malaysia, the United States of America and Brazil, with sampling period from January 3 to September 22, 2021. These samples were detected by SARS-CoV-2 antigen test kit (colloidal gold immunochromatography assay) and nucleic acid detection kit (real-time fluorescent quantitative reverse transcription polymerase chain reaction). Positive coincidence rates [(number of antigen-positive cases/nucleic acid-positive cases)×100%], negative coincidence rates [(number of antigen-negative cases/nucleic acid-negative cases)×100%], total coincidence rates [(number of cases with consistent results for both antigen and nucleic acid detection/number of total cases) ×100%], as well as Kappa values were calculated. The differences of the above indictors among different countries were evaluated by the coefficient of variation. The detection rates of the antigen test for samples with different cycle threshold values (Ct values) for the nucleic acid detection, different characteristics and different mutant strains were analyzed. Results: For all samples, the positive, negative, and total coincidence rate between the antigen test and nucleic acid assay was 90.8% (569/627), 99.7% (1 224/1 228) and 96.7% (1 793/1 855), respectively, and the consistency coefficient Kappa value was 0.924. Among these countries, the coefficient of variation for positive coincidence rates (except for Malaysia with a lot of samples with Ct value>30), negative coincidence rates (except for France without negative samples) and total coincidence rates (except for France) was 6%,<1%, and 6%, respectively. When Ct values were less than 25, the detection rates of antigen test were 83.3%-100% for each countries (the coefficient of variation was 6%); the total detection rate and the coefficient of variation was 93.4% (428/458) and 5%, respectively, for asymptomatic infected persons and cases within 7 days post onset of symptoms; the total detection rate for various SARS-CoV-2 mutant strains was 97.5% (119/122); and it showed negative results for samples from cases infected with other viruses, including influenza A virus subtype H1N1, influenza B virus, respiratory syncytial virus subgroups A and B, coxsackievirus 16, human metapneumovirus, parainfluenza virus types 1 and 4, Epstein-Barr virus and adenovirus. Conclusion: The SARS-CoV-2 antigen test kit showed excellent authenticity, and there were few differences for its indictors among nine countries, therefore it can meet the needs of large-scale early screening of SARS-CoV-2 infection.

[Value of proteinuria in assessing the severity of pre-eclampsia and its maternal and neonatal outcomes].

Objective: To investigate the value of proteinuria in evaluating the severity of pre-eclampsia (PE) and assessing the maternal and neonatal outcomes of PE. Methods: The clinical records of 265 pregnant women who were diagnosed with PE at Peking Union Medical College Hospital from January 2011 to June 2021 were retrospectively analyzed. According to 24-hour urine protein (24-hUPro) results, pregnant women were divided into two groups: the non-proteinuric group (24-hUPro<0.3 g, n=10) and proteinuric group (24-hUPro≥0.3 g, n=255). The proteinuric group was further divided into 3 subgroups based on proteinuria levels: mild group (0.3 g≤24-hUPro<2.0 g, n=119), moderate group (2.0 g≤24-hUPro<5.0 g, n=59), and severe group (24-hUPro≥5.0 g, n=77). The demographic and clinical data, laboratory indicators, pregnancy complications, maternal and neonatal outcomes were compared between different groups. Results: In proteinuric subgroups, increased proteinuria was associated with earlier onset gestations, higher incidence of headache, peripheral tissue edema, serosal effusion, intrauterine growth restriction, and abnormal umbilical cord blood flow (all P<0.05). There were no significant differences in the incidence of placental abruption, eclampsia and maternal mortality among the three subgroups, but there were significant differences in the incidence of neonatal birth weight and multiple neonatal complications (all P<0.05). Compared with the proteinuric group, the non-proteinuric group showed later onset gestation (median:34.7 vs 37.6 weeks) and gestational age of delivery (median:36.0 vs 38.4 weeks), lower proportion of ocular vascular lesions [56.7% (135/238) vs 2/9], higher birth weight (median: 2 325 vs 2 750 g), and lower rate of neonatal intensive care unit occupancy [54.3%(127/234) vs 1/10;all P<0.05]. Conclusions: The proteinuria plays an important role in assessing the severity of PE and maternal and neonatal outcomes, but it is not the only indicator. The non-proteinuric PE pregnant women might still lead to severe maternal and neonatal outcomes.

lnc94638 is a testis-specific long non-coding RNA involved in spermatozoa formation in Zeugodacus cucurbitae (Coquillett).

Long non-coding RNAs (lncRNAs) generally display tissue-specific distributions, and testis-specific lncRNAs form the highest proportion of lncRNAs in many species. Here, we presented a detailed analysis of testis-specific lncRNAs in the melon fly, Zeugodacus cucurbitae, a highly destructive insect pest of cucurbitaceous and other related crops. Most testis-specific lncRNAs were found to be long intergenic non-coding RNAs (lincRNA). The size distribution of these lncRNAs ranged between 600 and 1000 nucleotides. Testis-specific lncRNAs that harboured one isoform number and two exons were the most abundant. Compared to other male tissues, the testis had more highly expressed lncRNAs. The quantitative real-time polymerase chain reaction results of 10 randomly selected testis-specific lncRNAs showed expression patterns consistent with RNA-seq data. Further analysis of the most highly expressed testis-specific lncRNA, lnc94638, was undertaken. Fluorescent in situ hybridization assays localized lnc94638 to the apical region of the testis that contains mature spermatozoa. RNA interference-mediated knockdown of lnc94638 expression reduced spermatozoa numbers and impaired the fertility of Z. cucurbitae male. This study provides a catalogue of testis-specific lncRNAs, shows that the testis-specific lnc94638 is involved in spermatogenesis and has the potential to be used for treating male sterility.

[The 490th case: arthralgia, amenorrhea, aphasia].

A 25-year-old woman was admitted to Peking Union Medical Hospital presented with arthralgia for 5 years, amenorrhea for 16 months, and speech disorder for 3 months. This patient has been afflicted by intermittent pain in metacarpophalangeal and proximal interphalangeal joints of both hands for 5 years. Her menstruation has been irregular 1 year ago and rapidly progressed to amenorrhea. Laboratory tests revealed postmenopausal sex hormones levels (estradiol<5 ng/L, follicle-stimulating hormone 62.5 IU/L, luteinizing hormone 58.71 IU/L) and no antral follicles were seen in gynecologic ultrasound. She was diagnosed with premature ovarian failure and treated with hormone replacement therapy, still with no ovulation. Numbness and weakness of right arm has recurrently occurred to her 4 months ago, and persistent weakness of right limbs combined with motor speech disorder occurred 1 month later. Magnetic resonance angiography was suggestive of ischemic stroke. Hormone replacement therapy was discontinued. Comprehensive laboratory tests revealed positive anti-dsDNA, anti-SSA/SSB, anticardiolipin and anti-ß2GPⅠ antibodies. Systemic lupus erythematosus (SLE), antiphospholipid syndrome (APS) was diagnosed. Since no drug with gonadal toxicity had been applied to the patient before, her amenorrhea was considered to be due to autoimmune oophoritis secondary to SLE. After treated with high-dose glucocorticoid, mycophenolate mofetil and hydroxychloroquine for 4 months, her menstruation recurred and regularly occurred till now. In some cases, amenorrhea in SLE patient might be resulted from autoimmune oophoritis associated with lupus flare, instead of use of drug with gonadal toxicity.

Autism spectrum disorders: a meta-analysis of executive function.

Evidence of executive dysfunction in autism spectrum disorders (ASD) across development remains mixed and establishing its role is critical for guiding diagnosis and intervention. The primary objectives of this meta-analysis is to analyse executive function (EF) performance in ASD, the fractionation across EF subdomains, the clinical utility of EF measures and the influence of multiple moderators (for example, age, gender, diagnosis, measure characteristics). The Embase, Medline and PsychINFO databases were searched to identify peer-reviewed studies published since the inclusion of Autism in DSM-III (1980) up to end of June 2016 that compared EF in ASD with neurotypical controls. A random-effects model was used and moderators were tested using subgroup analysis. The primary outcome measure was Hedges' g effect size for EF and moderator factors. Clinical sensitivity was determined by the overlap percentage statistic (OL%). Results were reported according to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. A total of 235 studies comprising 14 081 participants were included (N, ASD=6816, Control=7265). A moderate overall effect size for reduced EF (Hedges' g=0.48, 95% confidence interval (CI) 0.43-0.53) was found with similar effect sizes across each domain. The majority of moderator comparisons were not significant although the overall effect of executive dysfunction has gradually reduced since the introduction of ASD. Only a small number of EF measures achieved clinical sensitivity. This study confirms a broad executive dysfunction in ASD that is relatively stable across development. The fractionation of executive dysfunction into individual subdomains was not supported, nor was diagnostic sensitivity. Development of feasible EF measures focussing on clinical sensitivity for diagnosis and treatment studies should be a priority.

[Expression of high-mobility group box-â and N-methyl-D-aspartate receptor in status epilepticus].

Objective: To investigate the expression changes of high mobility group box-1 (HMGB1) and the 2B receptor of N- methyl -D- aspartate receptor (NR2B) in status epilepticus (SE). Methods: (1) Primary hippocampal neurons from SD rats with 16 to 18 days of fetal age were cultured in vitro for 7 days, and exposed to Mg(2+) free media for 3 hours. Those cultured neurons were randomly divided into control group and intermittent hypoxia group. (2) SD rats with similar weight were selected and randomly divided into control group and SE model group. The rat model with SE was established by an intraperitoneal injection of lithium chloride-piloearpine (LI-PILO). Real-time PCR technique was used to detect the expression of HMGB1 and NR2B mRNA. Results: In Sombati's cell model cultured in normal concentration of oxygen, the HMGB1 mRNA expression levels were 0.005 01±0.000 54, 0.026 76±0.003 75, 0.003 52±0.000 33, and the NR2B mRNA expression levels were 0.008 84±0.000 69, 0.012 23±0.000 90, 0.029 11±0.000 71, respectively, at 2, 4 and 6 h; compared with the expressions of HMGB1 and NR2B mRNA at the same time points of Sombatis cell model groups, the differences were also significant (all P<0.05). After the successful establishment of epilepsy model, the HMGB1 mRNA expression levels were 0.000 11±0.000 09, 0.000 18±0.000 01, 0.000 11±0.000 01, and the NR2B mRNA expression levels were 0.196 12±0.009 41, 0.232 11±0.006 27, 0.272 48±0.005 84, respectively, at 6, 8 and 10 h; compared with the expressions of HMGB1 and NR2B mRNA at the same time points of control groups, the differences were all significant (all P<0.05). Conclusion: HMGB1 mRNA expression levels increase at 2, 4 h, decrease at 6 h in the Sombati's cell model in normal oxygen culture, while increase at 6, 8 h, and decrease at 10 h in LI-PILO induced rat model with SE; the NR2B mRNA relative expression increases with time in both the Sombati's cell model in normal oxygen culture and rat model of SE.

[Factors associated with syphilis/HIV infection among men who have sex with men in Shenzhen,2011-2016].

Objective: To investigate the factors associated with syphilis/HIV infection among men who have sex with men (MSM) maintaining a single sexual partner in Shenzhen. Methods: Respondent driven sampling and snowball sampling method were used to recruit MSM receiving voluntary counseling and testing in Shenzhen Rainbow clinic from 2011 to 2016. INCLUSION CRITERIA: aged 18 years and above; reported having one or more anal sexual partners in the recent 6 months. A total of 3 109 men who have sex with men (MSM) were involved in the study. Questionnaire-based interviews were conducted on a one-on-one basis. Data were collected including socio-demographic information, human immunodeficiency virus (HIV) testing history, history of blood donation and drug abuse in the recent two years, self-identified sexual orientation, role in homosexual behavior, ever being money boys (MB) and clients of MB, female sexual partners in the recent 6 months. 5 ml blood samples were taken after questionnaires. Syphilis was screened using toluidine red unheated serum test for the antibody of treponema pallidum and then confirmed by treponema pallidum particle assay. The enzyme-linked immunosorbent assay (ELISA) was used for screening HIV and western blot was used to confirm the HIV screening results. The difference of condom use among MSM between female sexual partners and male sexual partners were compared. Multivariate unconditional stepwise logistic regression model was used to analyze factors associated with syphilis infection and HIV positive among MSM. Results: A total of 3 109 eligible participants with mean (SD) age of 31.49 (8.64) years were enrolled in this study. Among them, 565 cases were infected with syphilis (18.17%), 330 cases were infected with HIV (10.61%), and 165 cases (5.31%) were syphilis co-infected with HIV. 791 (25.44%) reported maintaining a single sexual partners in the recent 6 months. Compared to MSM with multiple sexual partners, the prevalence of syphilis and HIV infection among MSM with a single sexual partner were lower, and the OR (95%CI) were 0.64 (0.51-0.81) and 0.66 (0.49-0.90), respectively. Conclusion: The proportion of fixed single sexual partner was much lower among MSM in Shenzhen. Maintaining a single sexual partners can reduce the risk of syphilis/HIV infection among MSM.

[Analysis of curative effect of surgical therapy for male prolactinoma].

OBJECTIVE: To summarize the treatment outcome of surgery for male prolactinoma. METHODS: One hundred and eighty-four cases of male prolactinoma who underwent surgery were analyzed retrospectively.The average age of patients was 36.3 years, and the mean course was 41.9 months.The main clinical manifestation included sexual dysfunction (47.4%), headache (55.9%) and visual disturbance (46.7%). The serum prolactin levels ranged from 6.83 to 9 325.32 nmol/L.The size of tumors varied from 6 mm to 70 mm.Suprasellar adenoma with visual deficits accounted for 40.7%. 98.9% patients underwent pituitary adenoma resection via single nasal transsphenoidal approach. RESULTS: Postoperative pathological Ki-67 index of most patients (45.1%) were less than 1%.After surgical therapy, 163 patients (88.6%) got relief of symptoms, 57 patients (31.0%) achieved initial remission, and 26 patients (45.6%) unfortunately recurred. The possibility of gross resection decreased as tumor size increased (P<0.05). Preoperative PRL had no significant relation with both tumor size and extent of resection (P>0.05). The favorable prognosis predictive factors included microadenoma, intrasellar adenoma and Ki-67 index≤3 (P<0.01). CONCLUSION: Male prolactinomas is an invasive tumor with higher serum prolactin level.Tumor size could predict degree of gross resection while predictors for recurrence include tumor size, growth pattern and Ki-67 index.

[Factors associated with commercial sexual behavior among men who have sex with men in Shenzhen, China, in 2011-2015].

Objective: To investigate the status and factors associated with commercial sexual behavior among men who have sex with men (MSM) in Shenzhen. Methods: A convenience sampling method was used to recruit MSM in Shenzhen from 2011 to 2015. Questionnaire-based interviews were conducted on a one-on-one basis. Data were collected, including socio-demographic information, HIV testing history, history of blood donation and drug abuse in the last 2 years, self-reported sexual orientation, role in homosexual behavior, and experience serving as a male sex worker and/or as a client of male sex workers. Blood samples (5 ml) were taken and tested for treponema pallidum and HIV antibodies. The rate of MSM serving as male sex workers among different age groups was analyzed using the Cochran-Armitage trend test. Factors associated with commercial sexual behavior were analyzed by univariate logistic regression and multivariate unconditional logistic regression. Results: Among the 3 040 MSM recruited, 341 (11.2%) reported having served as male sex worker. The prevalence rates of syphilis, HIV, and syphilis-HIV co-infection among all recruited MSM were 18.3% (556/3 040), 9.8% (297/3 040), and 5.1% (154/3 040), respectively. The prevalence rates of syphilis, HIV, and syphilis-HIV co-infection among those who served as male sex worker were 27.0% (92/341), 16.4% (56/341), and 8.8% (30/341), respectively, and the prevalence rates among MSM with no experience as male sex worker were 17.2% (464/2 699), 8.9% (241/2 699), and 4.6% (124/2 699), respectively. Compared with non-male sex worker MSM, male sex worker had a significantly higher prevalence rates of syphilis, HIV, and syphilis-HIV co-infection (with χ2 values of 19.41, 19.28, and 11.12, and P-values of <0.001,<0.001, 0.001, respectively). The results of the multivariate logistic regression analysis indicate that>30-year-old MSM or education level of college or above, reside in Shenzhen for 0.5-3.0 years or the living years above 3.0 years are less likely to serve as male sex worker compared with<30-year-old MSM, education level of high school or below, who reside in Shenzhen for less than 0.5 years; the OR (95%CI) values were 0.55 (0.42-0.71), 0.10 (0.06-0.14), 0.46 (0.31-0.68), and 0.23 (0.16-0.33), respectively. MSM having a monthly income of >5 000 yuan, drug abuse history, experience as male sex worker clients, and self-reported insertive and receptive anal sex behaviors were more likely to have experience serving as male sex worker compared with those having a monthly income of <3 000 yuan (OR=2.57, 95%CI: 1.85-3.57), no drug abuse history (OR=3.23, 95% CI: 2.14-4.87), no experience as male sex worker clients (OR =1.50, 95% CI: 1.04-2.15) and who engage in predominantly insertive anal sex behaviors (OR=1.77, 95% CI: 1.34-2.35). Conclusion: Age, education level, duration of residence in Shenzhen, monthly income, history of drug abuse, role in homosexual activity, and had experience commercial sex activity were associated with serving as a male sex worker among MSM. These factors need to be considered when designing syphilis/HIV prevention programs for MSM.

[Update on pubertal development among primary school students in Shanghai, 2014].

Objective: To investigate the current prevalence of pubertal development in healthy Shanghai schoolchildren. Methods: This study was a cross-sectional investigation focused on current pubertal development conducted in healthy Shanghai schoolchildren by multi-stage cluster sampling. The sample included 17 571 children in grades 1-5 investigated in June 2014. The data were weighted by inverse probability weighting (IPW) to make them more representative. At examination, stages of breast and pubic hair development were rated according to the Tanner method. Testicular volume was determined. Data on menarche and spermatorrhea were collected by the status quo method. The rates of precocious puberty, breast, and pubic hair development of Tanner stage ≥Ⅱ in girls aged 6-7 years, menarche in girls aged 6-9 years, and testicular volume ≥4 ml and pubic hair development of Tanner stage ≥Ⅱ in boys aged 6-8 years were calculated. All the data were weighted by IPW. Results: After data processing, 16 197 children's data were analyzed. In girls aged 6-7 years, 17.2% and 2.5% showed evidence of breast and pubic hair development at Tanner stage ≥Ⅱ, respectively. In girls aged 6-9 years, 0.3% had experienced menarche. Schoolgirls' rate of menarche was 4.7%. In girls aged 6-7 years, 19.0% were diagnosed with precocious puberty according to the classic criteria. In boys aged 6-8 years, 1.7% had testicular volume ≥4 ml, and 0.6% showed evidence of pubic hair development at Tanner stage ≥Ⅱ. Schoolboys' incidence rate of spermatorrhea was 0.1%. In boys aged 6-8 years, 2.3% were diagnosed with precocious puberty according to the classic criteria. All the numbers above were weighted. Conclusion: Proper education on adolescence and sex is essential for Shanghai schoolchildren.

Regulatory functions of docosahexaenoic acid on ion channels in rat ventricular myocytes.

OBJECTIVE: The aim of this study was to study the regulatory functions of docosahexaenoic acid (DHA) on resting potential (RP), action potential duration (APD), delayed rectifier potassium current (Ik), and inwardly rectifier potassium current (Ik1) in rat ventricular myocytes, and analyze the related anti-arrhythmia mechanism. MATERIALS AND METHODS: Rat ventricular myocytes were isolated by enzyme digestion method. RP, APD, Ik and Ik1 in individual ventricular myocytes were recorded by patch-clamp technique with whole-cell configuration. Effects of DHA with various concentrations (0, 20, 40, 60, 80, 100 and 120 mmol/L, respectively) on RP, AP, Ik and Ik1 were investigated. RESULTS: There was no statistical difference of RP with different DHA concentrations (p > 0.05, n = 20), and the 25%, 50% and 90% of APD (APD25, APD50, and APD90) were gradually prolonged with increase of DHA concentration, respectively (p < 0.05, n = 20). IK gradually blocked and the I-V curve was downward shifted, according to increase of DHA concentration (p < 0.05, n = 20). The DHA half effect concentration (EC50) was 47.52 ± 2.32 µmol/L. With increasing DHA concentration, the steady-state inactivation curve shifted to left, and the recovery curve shifted to right. DHA had no significant effect on IK1 (p > 0.05, n = 20). CONCLUSIONS: DHA has regulatory functions on RP, APD, Ik and Ik1 in rat ventricular myocytes, which may be one of the related antiarrhythmic mechanisms.

Impact of MTHFR polymorphisms on methylation of MGMT in glioma patients from Northeast China with different folate levels.

Hypomethylation of the O6-methylguanine-DNA-methyltransferase (MGMT) promoter in glioma cells has been associated with temozolomide resistance. S-adenosylmethionine (SAM), which is produced during folate metabolism, is the main source of methyl groups during DNA methylation. As a key enzyme during folate metabolism, polymorphisms of 5,10-methylenetetrahydrofolate reductase (MTHFR) may regulate folate end-products. We investigated the effect of typical polymorphisms of MTHFR (C677T and A1298C) on MGMT methylation based on different serum folate levels in patients with glioma from Northeast China. A total of 275 patients with glioma and 329 without malignant tumors were tested. Serum folate concentration was assayed by using the electrochemiluminescence immunoassay. MTHFR polymorphisms were detected by Taqman-Fluorescence quantitative polymerase chain reaction (PCR). Methylation-specific PCR was used to assess MGMT methylation. The constituent ratio of glioma patients below the serum folate biological reference value was significantly higher than that of the control population (P < 0.001). In patients with oligodendroglioma and glioblastoma, heterozygotes for the A1298C mutation were found in higher frequency than homozygotes or wild types (oligodendroglioma, P < 0.001; glioblastoma, P < 0.01). When grouped by the median or biological reference value of serum folate, only homozygotes for C677T with low levels of folate were significantly associated with decreased methylation of MGMT (median, P < 0.001; biological reference value, P = 0.036). These data suggest that, in combination with a negative folate balance in glioma patients, T/T genotypes in MTHFR C677T may be associated with MGMT demethylation.

Evaluation of the efficacy of immunocastration vaccine composed of gonadotrophin-releasing hormone conjugated with Salmonella typhimurium flagellin in rats.

Immunocastration is an alternative method to replace surgical castration that is commonly performed in domestic and pet animals. In this study, a new immunocastration vaccine was developed, and its efficacy was evaluated in male rats. Six tandem copies of gonadotrophin-releasing hormone (GnRH) peptide were genetically fused to Salmonella typhimurium flagellin fljB (STF2) that is a ligand of toll-like receptor 5 (TLR5). The recombinant STF2-GnRH protein expressed in Escherichia coli was used as the immunocastration vaccine. Sixteen male rats were equally assigned to four groups. Excluding the control rats, three groups were immunized with 100, 200 and 400 µg of the STF2-GnRH vaccine, respectively. All of the immunized rats developed significantly higher titres of antibodies to GnRH than the control rats. The size and weight of both testes and epididymides from the immunized rats were significantly smaller than those of the control rats. Testicular tissues in the immunized rats demonstrated atrophy of seminiferous tubules and decreased numbers of both spermatogonia and spermatocytes. These data indicate that the newly developed STF2-GnRH vaccine has a potent immunogenicity to GnRH and efficiently suppresses the development of testes in rats.

Correlations of IL-18 and IL-6 gene polymorphisms and expression levels with onset of glioma.

OBJECTIVE:   The aim of this study was to explore the correlations of interleukin (IL)-18 and IL-6 gene polymorphisms and expression levels with the onset of glioma. PATIENTS AND METHODS: The differences in the expression levels of IL-18 and IL-6 between glioma patients and normal people in the Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases were analyzed. A total of 200 glioma patients and 200 healthy people were taken as the research subjects. Peripheral blood was collected to extract deoxyribonucleic acids (DNAs). IL-18 and IL-6 gene polymorphisms were detected and analyzed combined with haplotype analysis and gene expression levels of IL-18 and IL-6, as well as their levels in serum. RESULTS: Both IL-18 and IL-6 were highly expressed in tumor tissues of glioma patients, whereas they were lowly expressed in normal cerebral tissues, with statistically significant differences (p<0.05). Statistically significant differences in the allele distributions of IL-18 gene polymorphisms rs371411440 (p=0.041) and rs371828055 (p=0.002) and IL-6 gene polymorphisms rs201211345 (p=0.000) and rs201439472 (p=0.003) were observed between disease group and control group (p<0.05). Genotype distributions of IL-18 gene polymorphism rs371828055 (p=0.005) and IL-6 gene polymorphisms rs201211345 (p=0.000) and rs201439472 (p=0.019) in disease group were significantly different from those in control group (p<0.05). Disease group exhibited significantly higher frequencies of genotype GG of IL-18 gene polymorphism rs371828055, genotype AA of IL-6 gene polymorphism rs201211345 and genotype TT of IL-6 gene polymorphism rs201439472 than control group (p<0.05). There were statistically significant differences in the distributions of the dominant model AA+AC of IL-6 gene polymorphism rs201211345 (p=0.016) and the recessive model GT+TT of IL-18 gene polymorphism rs371828055 (p=0.010) between the two groups (p<0.05). Differences in the distributions of haplotypes CC (p=0.001) and GT (p=0.027) of IL-18 gene polymorphisms rs371411440 and rs371828055 and haplotypes AC (p=0.009), AT (p=0.000) and CT (p=0.000) of IL-6 gene polymorphisms rs201211345 and rs201439472 were observed between disease group and control group (p<0.05). In addition, a high degree of linkage disequilibrium was detected between IL-6 gene polymorphisms rs201211345 and rs201439472 (D'=0.583). The genotypes of IL-18 gene polymorphism rs371828055 were evidently correlated with the gene expression of IL-18 (p=0.000). Meanwhile, patients with genotype GT had a distinctly lower expression level of IL-18 (p<0.05). The genotypes of IL-6 gene polymorphism rs201211345 were obviously associated with the expression of IL-6 (p=0.002). The expression of IL-6 was markedly down-regulated in patients carrying genotype AA (p<0.05). Consistent with the expression levels of IL-18 and IL-6, the genotypes of IL-18 gene polymorphism rs371828055 were associated with the content of serum IL-18 (p<0.05). Moreover, patients carrying genotype GT had distinctly lower content of serum IL-18 (p<0.05). Additionally, the genotypes of IL-6 gene polymorphism rs201211345 were evidently correlated with the content of serum IL-6 (p<0.05), and the content of serum IL-6 declined distinctly in patients with genotype AA (p<0.05). CONCLUSIONS: IL-18 and IL-6 gene polymorphisms and expression levels are significantly correlated with the onset of glioma.

Polar nanodomains and giant converse magnetoelectric effect in charge-ordered Fe2OBO3.

The magnetoelectric coupling and polar nanodomains in the charge-ordered Fe2OBO3 have been extensively studied from room temperature down to 100 K. In situ TEM investigations demonstrate that the charge-ordering transition characterized by an incommensurate modulation could evidently result in remarkable polar nanodomains at low temperatures. This kind of nanodomain could play a critical role in triggering a high dielectric constant and notable dielectric dispersion as observed in Fe2OBO3. Moreover, measurements of the magnetoelectric coupling under electrical field demonstrate the existence of giant electrically induced changes in magnetization around the magnetic transition.

Diabetes Medication Assistance Service Stage 1: impact and sustainability of glycaemic and lipids control in patients with Type 2 diabetes.

AIMS: To investigate (i) optimal intensity (four visits vs. six visits) and duration (6 vs. 12 months) of the Diabetes Medication Assistance Service in community pharmacy and (ii) sustainability of improvements in patients' diabetes control associated with differing intensities of intervention. METHODS: A national quota sample of 90 community pharmacies in Australia were randomly assigned into group 1 (6-month Diabetes Medication Assistance Service) or group 2 (12-month Diabetes Medication Assistance Service) and subsequently recruited a total of 524 patients. A wide range of clinical (HbA(1c) , blood pressure, lipids) and quality-of-life outcome measures were assessed. RESULTS: The 6- and 12-month Diabetes Medication Assistance Service resulted in significant and similar reductions in HbA(1c) (-0.9 mmol/mol; 95% CI -0.7 to -1.1) -, total cholesterol (-0.3 mmol/l; 95% CI -0.1 to -0.4) and triglycerides (-0.3 mmol/l; 95% CI -0.1 to -0.5). There was also a significant reduction in the number of patients who were at risk of having a cardiovascular event in the next 10 years. For the subset of patients for whom data were available at baseline, completion and 18 months, improvements in HbA(1c) and total cholesterol were sustained at 18 months and triglycerides showed a further improvement at 18 months. CONCLUSIONS: The Diabetes Medication Assistance Service resulted in significant improvements in diabetes control that were independent of intensity and duration of the service and showed evidence of being sustained at 18 months. The extent and sustainability of clinical improvements achieved by the Diabetes Medication Assistance Service, together with the resulting reduction in cardiovascular risk, should translate into future cost savings to healthcare systems by delaying and reducing diabetes-related complications.