OBJECTIVESince glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most cases have single nucleotide mutations in the coding region, and current test methods for gene mutation have some missed detections, this study aimed to investigate the feasibility of RT-PCR sequencing in the detection of gene mutation in G6PD deficiency.

METHODSAccording to the G6PD/6GPD ratio, 195 children with anemia of unknown cause or who underwent physical examination between August 2013 and July 2014 were classified into G6PD-deficiency group with 130 children (G6PD/6GPD ratio <1.00) and control group with 65 children (G6PD/6GPD ratio≥1.00). The primer design and PCR amplification conditions were optimized, and RT-PCR sequencing was used to analyze the complete coding sequence and verify the genomic DNA sequence in the two groups.

RESULTSIn the G6PD-deficiency group, the detection rate of gene mutation was 100% and 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 boys; 13 heterozygous missense mutations, 1 homozygous same-sense mutation (C1191T) which had not been reported in China and abroad, and 14 single nucleotide polymorphisms of C1311T were detected in 37 girls. The control group showed a high rate of missed detection of G6PD deficiency (carriers) in the specimens from girls (35%, 13/37).

CONCLUSIONSRT-PCR sequencing has a high detection rate of G6PD gene mutation and a certain value in clinical diagnosis of G6PD deficiency.

Adolescent ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Infant ; Male ; Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Objective To investigate the feasibility of reducing bolus?tracking monitor frequency in coronary CT angiography (CTA). Methods This prospective study including 120 patients with suspected coronary artery disease (CAD). According to the examination registration order, the patients were divided into groups A, B and C (n=40 for each group). All patients underwent coronary CTA with bolus?tracking technology, and were monitored at 10 s after the injection of contrast. The monitoring frequency of bolus?tracking for Group A was every 1.14 s, that for Group B was every 1.47 s , and for Group C was every 2.00 s, while the trigger threshold was set as 100 HU. To evaluate the image quality, the objective evaluation included signal noise ratio (SNR) and contrast noise ratio (CNR) of aorta (AO), CNR of left main coronary artery (LM) and right coronary artery (RCA), and the subjective score was recorded for each coronary artery segment. The monitoring times when CT density of the region of interest (ROI) reached the threshold, the CT value and the effective dose (ED) in the 3 groups were recorded. Objective image quality, monitoring parameters and radiation dose were compared using analysis of variance method, subjective image quality was compared withχ2 tests. Results There was no significant difference in AO (SNR and CNR), LM (CNR) and RCA (CNR) among the 3 groups, respectively (P>0.05). Subjective image quality scores of groups A, B, C were (1.879±0.042), (1.876±0.042), (1.881±0.042 ), with no significant difference (χ2=0.003,P>0.05). The monitoring times of to reach the threshold in groups A, B, C were (4.78±2.37), (3.76±1.39), (2.77±0.99), and ED were (0.058±0.031),(0.031±0.011), (0.021±0.007) mSv, with the significant difference (F=9.009, 31.998, respectively, P<0.01). Peak CT values during monitoring among three groups were (133 ± 24), (142 ± 39), (137±26) HU, respectively, with no significant difference (F=0.575,P=0.565). Conclusions It is feasible to reduce monitoring times when performing coronary CTA in dual?source CT scanner. The bolus?tracking monitor frequency in every 2 seconds can not only obtain satisfactory image quality, but also significantly reduce radiation dose.

Objective To investigate the impact of tube current and tube voltage on CT attenuation,the correlation between CT attenuation and iodine concentration,as well as the percentage of reducing dosage for contrast agent while reducing the tube voltage.Methods A total of 100 saline solutions with decreasing dilution of contrast medium,in which concentration was between 0.5 to 50.0 mg/ml with the interval of 0.5 mg/ml,was produced.Each of the 25 syringes with a 4 ml sample was fixed on a cylindrical CT calibrated water phantom with an equal distance used the tape.CT scans were performed with a total of 15 scanning methods of the combination of the different tube voltages (70,80,100,140 kV) and tube current (100,200,280 mA).All of the CT attenuations were measured and recorded.The differences of CT attenuations under different scanning tube currents and tube voltages were compared with one-way ANOVA.The Pearson correlation analysis was used to analyze the relationship between CT attenuation and iodine concentration,and linear correlation equations were calculated and shown by regression analysis.According to the equations,the changes of contrast medium dosage were calculated with the changes of tube voltage.Results There was no significant difference in CT attenuations on different tube currents when the tube voltage was fixed (all P＞0.05),while when the tube current was fixed,the difference of CT attenuations on different tube voltages was statistically significant (all P＜0.05).Under different scanning conditions,the CT attenuations was linearly related to the iodine concentration (r2 was 0.953 to 0.997,all P＜0.01).While the tube voltage was reduced from 140 kV to 120 kV,100 kV,80 kV,70 kV,respectively,the iodine concentration of the samples were reduced by 15.4％,33.7％,53.4％,64.7％ respectively.While the voltage was reduced from 120 kV to 100 kV,80 kV,70 kV,respectively,the iodine concentration were rednced by 21.6％,44.9％,58.2％,respectively.While the voltage was reduced from 100 kV to 80 kV and 70 kV,the iodine concentration was reduced by 29.7％ and 46.7％,respectively.While the voltage was reduced from 80 kV to 70 kV,the iodine concentration was reduced by 24.1％.Conclusion CT attenuation can keep constant in low tube voltage setting by reducing the dosage of contrast agent,which can achieve a low radiation dose and low contrast agent dosage.

Carcinoma ; diagnosis ; pathology ; Humans ; Laryngeal Neoplasms ; diagnosis ; pathology ; Larynx ; pathology

Objective: To investigate the status of knowledge about acquired immune deficiency syndrome(AIDS) among college freshmen in Jiangsu Province, and to analyze the influences of AIDS-related knowledge on their sexual attitudes and behaviors. Methods: A total of 8 633 freshmen from 20 colleges and universities in Jiangsu Province were enrolled by multi-stage stratified sampling. The questionnaire for young students about the eight items of AIDS basic knowledge were used to investigate the AIDS-related knowledge, sexual attitudes and behaviors. Multivariate Logistic regression model was used to analyze the influencing factors of the AIDS-related knowledge and to adjust the influences of AIDS-related knowledge on sexual attitudes and behaviors. Results: The overall qualified rate of AIDS-related knowledge among college freshmen in Jiangsu Province was 87.6% and the qualified rate of 8 items were between 59.8% and 98.3%. The types of schools, nationality, mother s education level and whether only child or not were identified as independent factors for qualified rate of AIDS-related knowledge(P<0.05). The awareness of AIDS-related knowledge had significant impacts on the sexual attitudes and behaviors of college freshmen. Students with insufficient knowledge were more likely to approve premarital sexual behaviors(OR=1.31, 95%CI=1.11-1.63), willing to use condoms consistently(OR=1.67, 95%CI=1.43-1.92) and to accept the HIV tests(OR=4.54, 95%CI=3.13-6.52). Students with unqualified knowledge were more likely to have sexual behavior(OR=1.89, 95%CI=1.42-2.49), while those with sexual experiences and insufficient AIDS knowledge were more likely to use condoms consistently(OR=2.13, 95%CI=1.22-3.78). Conclusion The education on AIDS-related knowledge for teenagers was helpful to construct correct sexual concepts and develop healthy sexual behaviors. However, the AIDS-related knowledge of college freshmen in Jiangsu Province is not sufficient and needed to improve.

Objective To evaluate the effects of turmeric volatile oil (TVO) combined with cisplatin on the proliferation and apoptosis of a human cutaneous squamous cell carcinoma cell line A431,and to explore their mechanisms.Methods Some cultured A431 cells at exponential growth phase were divided into several groups to be treated with 5,10,20,40 and 80 mg/L TVO,as well as high-glucose Dulbecco's modified Eagle's medium (DMEM) containing 1％ dimethyl sulfoxide (DMSO,control group),respectively.After 24-hour treatment,cell counting kit 8 (CCK8) assay was performed to estimate the proliferative activity of A431 cells in the above groups.Some other A431 cells were divided into 4 groups:control group treated with high-glucose DMEM containing 1％ DMSO,TVO group treated with 40 mg/LTVO,cisplatin group treated with 10 mg/L cisplatin,and TVO + cisplatin group treated with 40 mg/L TVO and 10 mg/L cisplatin.After 24-hour treatment,CCK8 assay was performed to estimate the cellular proliferative activity,inverted microscopy to observe changes in cell morphology,fluorescence microscopy to detect cell apoptosis after acridine orange (AO)/ethidium bromide (EB) double-staining,colorimetry to evaluate the activity of Caspase-3 and Caspase-9,and Western blot analysis to determine the protein expression of Caspase-3 and p-glycoprotein.Results After 24-hour treatment with 5,10,20,40 and 80 mg/L TVO,the cell proliferation rates were inhibited by (12.83 ± 6.4)％,(16.27 ± 11.4)％,(21.61 ± 9.1)％,(33.11 ± 2.0)％ and (46.00 ± 3.3)％ respectively,and the inhibition rates were all significantly higher in these groups than in the control group (4.03％ ± 1.4％,all P ＜ 0.05).The 50％ inhibitory concentration (IC50) of TVO at 24 hours was (61.66 ± 1.03) mg/L.Compared with the control group,the proliferation inhibition rates significantly increased in the TVO group,cisplatin group and TVO + cisplatin group (all P ＜ 0.05),suggesting that the combination of TVO and cisplatin showed synergistic inhibitory effects with a combination index of 1.366.Moreover,A431 cells turned round to different extents and became apoptotic in the TVO group and cisplatin group,and the TVO + cisplatin group showed obviously decreased number of cells and a large number of cell debris.The TVO + cisplatin group also showed significantly increased activity of Caspase-3 (1.520 ± 0.115) and Caspase-9 (2.760 ± 0.297) as well as protein expression of Caspase-3 (1.482 ± 0.016) compared with the TVO group (Caspase-3 activity:1.117 ± 0.095;Caspase-9 activity:1.259 ± 0.059;Caspase-3 protein expression:1.156 ± 0.006,all P ＜ 0.01) and cisplatin group (Caspase-3 activity:1.381 ± 0.089;Caspase-9 activity:1.829 ± 0.171;Caspase-3 protein expression:1.296 ± 0.021,all P ＜ 0.01),but significantly decreased p-glycoprotein expression (0.528 ± 0.014) compared with the TVO group (1.311 ± 0.011,P ＜ 0.01) and cisplatin group (1.169 ± 0.012,P ＜ 0.01).Conclusion TVO combined with cisplatin can synergistically inhibit the proliferation of A431 cells and induce cell apoptosis,which may be associated with activation of the caspase system and decreased expression of pglycoprotein.

Aim To investigate the role of P2X7 recep-tor and its mediated NLRP3 inflammatory signaling pathway in alcohol-induced liver injury. Methods The acute alcoholic liver injury model was established by NIAAA method, and thirty C57BL/6 male mice were randomly divided into three groups (n =10):control group, model group, A438079 group, The three groups were processed as follows in the last week:control group and model group: given an equal dose of saline intraperitoneal injection(about 0.2 mL/only) once a day. According to the weight of the mice, A438079 group was given intraperitoneally injection by 200 μmol·kg-1of A-438079 (prepared at 7 g·L-1 of A438079,about 0.2 mL/only) once a day. And it was given a single 31.5% alcohol solution by intragas-tric administration on the last day of the morning,with the dose of 10 mL·kg-1. Nine hours later alanine aminotransferase (ALT), aspartate aminotransferase (AST),cholesterol(TCHO),triglyceride(TG) were measured by orbital blood in mice. HE staining was used to observe the pathological changes of the liver. Immunohistochemical method was applied to detect the expression of P2X7R in liver tissues. Western blot was employed to detect the levels of P2X7R, NLRP3, ASC, IL-1β and IL-18 in liver tissues. Results Compared with control group,the levels of ALT,AST, TG and TCHO in model group were significantly en-hanced, and the liver injury was obvious. Compared with model group, the levels of ALT, AST, TG and TCHO in A438079 group significantly decreased. Compared with control group, the expressions of P2X7, NLRP3, ASC, IL-1β, IL-18 in model group were significantly higher than those in control group. Compared with model group, the expression levels of P2X7, NLRP3, ASC, IL-1β and IL-18 in A438079 group significantly decreased. Conclusion Alcohol-induced liver injury may be associated with P2X7R-NLRP3 signaling pathway.

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.

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.

Objective:To explore the profile and gene functional changes of gut microbiota (GM) in women with postmenopausal osteoporosis (PMOP) in Northwest China, and the correlations between GM and bone mineral density (BMD).Methods:From November 2018 to October 2019, postmenopausal women were screened on their initial visits to our hospital, and 24 new osteoporosis (OP) patients, 30 new osteopenia patients and nine negative controls were recruited. Fecal samples were collected for GM DNA extraction, and Illumina platforms were used for high-throughput sequencing of 16S rRNA and metagenome. Species annotation, GM profile and gene functions were viewed and analyzed.Results:GM profiles were significatly different in different groups, and the LDA scores of Peptostreptococcaceae, Romboutsia, unidentified Clostridiales, Megamonas, Erysipelatoclostridium, Klebsiella and Erysipelatoclostridium ramosum were more than 3 in OP group. Metagenomic sequencing analysis indicated that gene numbers were positively correlated with BMD, and metabolism, carbohydrate metabolism, starch and sucrose metabolism and oxidative phosphorylation were negatively correlated with BMD. Receiver operating characteristic curve(ROC) showed that carbohydrate metabolism, starch and sucrose metabolism, amino sugar and nucleotide sugar metabolism, oxidative phosphorylation, respectively, could identify OP with preferable sensitivity and specificity (areas under curve were 0.70, 0.72, 0.73 and 0.75, respectively). Conclusions:High-throughput sequencing had great potential for GM analysis of postmenopausal women with OP, providing evidence of the correlations between GM and BMD.