Abstract: Objective　To construct HepG2, Huh7 cell lines stably express hepatitis B virus X (HBx) mutant (C1653T, T1753C), and explore their effect on the biological behavior of hepatocellular carcinoma cells. Methods　The lentivirus plasmid of pLVX-HBxC1653T-IRES-tdTomato, pLVX-HBxT1753C-IRES-tdTomato were obtained by PCR site mutagenesis according to wild type ayr HBx. Double enzyme digestion and Sanger sequencing were performed for accuracy of plasmid. Blank HepG2 and Huh7 cells were used as the control group, HepG2, Huh7 cells were infected by pLVX-HBx-IRES-tdTomato, pLVX-HBxC1653T-IRES-tdTomato, and pLVX-HBxT1753C-IRES-tdTomato lentivirus solution, then monoclonal cell was selected by 0.6 μg/mL puromycin. Immunostaining and Western Blot were performed for the verification of stable strains. CCK8 assay was performed for the proliferation capacity of stable strains. Western Blot was performed for expression of EMT-related signal molecules in cells. The independent samples t-test was used for comparison between two groups. Results Double enzyme digestion and Sanger sequencing showed that that the size of the cut fragments of recombinant lentiviral plasmids was correct, and the point mutation location and base substitution were correct, suggesting that the plasmid of pLVX-HBx-IRES-tdTomato, pLVX-HBxC1653T-IRES-tdTomato, pLVX-HBxT1753C-IRES-tdTomato were constructed successfully. Immunostaining and Western blot showed that HBX were expressed in stable strains, while there was no HBX expression in the blank control group, indicating that the HepG2 and Huh7 cell lines stably expressing HBx, HBxC1653T, HBxT1753C were successfully constructed. CCK8 assay showed that the proliferation capacity of HBx and mutant were enhanced compared to the control group (P<0.01), HBx C1653T displayed further additive the effect compared to HBx (P<0.05). Moreover, HBxC1653T mutation also significantly upregulated N-cadherin expression and downregulated E-cadherin expression, thus promoting the occurrence of EMT. Conclusions　HepG2 and Huh7 cell lines stably expressing HBx, HBxC1653T, HBxT1753C were successfully constructed, HBxC1653T mutation significantly enhanced the proliferation of HCC cells and epithelial to mesenchymal transition occurrence.

Objective To explore rules of changes in velocity of ultrasound wave transmission at the tibia and radius by a quantitative bone mineral density ultrasound scanner in examination of early incipient rickets in infants and its significance in clinical diagnosis.Methods One hundred and fifty-seven infants who visited child health-care clinic of the Capital Institute of Pediatrics,Beijing during May 2004 to December 2007 and clinically diagnosed as rickets according to the Criteria of Diagnosis for Rickets in Infants and Young Children formulated in 1986 by the Ministry of Health,as well as 124 normal healthy infants as controls,were enrolled in the study and followed-up for three months.Velocity of ultrasound wave transmission at the tibia and radius in all the infants were measured by a bone mineral density ultrasound scanner (Sunlight Omnisense 7000R made in Israel).Results One hundred and fifty-seven infants were clinically diagnosed as rickets,127 of them undergone with carpal plane roentgenography and 90 of the 127 with positive change in bone x-ray imaging and 37 without it.Velocity of ultrasound wave transmission at the tibia and radius measured by z-score was significantly lower in infants with clinically diagnosed rickets than that in healthy controls (Z-values of-10.411 and-10.399 at the tibia and-5.646 and-5.517 at the radius,respectively,P = 0.000 with Mann-Whitney test).Velocity of ultrasound wave transmission at the tibia and radius measured by z-score was lower in those with positive change in x-my imaging than that in those without it,but not reaching a level of statistically significant difference.Velocity of ultrasound wave transmission and z-score correlated positively with change in bone x-ray imaging,respectively with Spearman coefficients of correlation of 0.581 and 0.677 for tibia,0.316 and 0.467 for radius (P = 0.000).Receiver operating characteristic (ROC) curve was drawn from those with rickets and positive left carpal plane roetgenograph.Area under curve of ROC for z-score of velocity of ultrasound wave transmission at the tibia was 0.812 and 0.799 (95% CI 0.758-0.856 and 0.742-0.855,P =0.000),respectively.Area under ROC curve of z-score of velocity of ultrasound wave transmission at the radius was 0.715 and 0.697 (95% CI 0.650-0.780,0.631-0.764,P =0.000),respectively.Cut-off value of z-score was-205--1.95 at the tibia,according to the largest sensitivity and the least false-positivity,with sensibility of 0.8 to 0.9 and specificity of 0.733 to 0.702.As its sensitivity at the radius was more than 0.7,its specificity was lower than 0.524.Conclusions Quantitative ultrasound scanning can be used to detect insufficient bone mineral density in infants and their early skeletal change by rickets but without change in bone x-ray imaging.Velocity of ultrasound wave transmission at the tibia can be used as an indicator to predict early rickets in infants rather than its severity.

Percutaneous coronary intervention is an extremely effective method for the treatment of acute myocardial infarction.In some patients, allergic reactions to metals such as in-stent restenosis and coronary aneurysm dilatation will occur.Currently, treatment options for such patients remain controversial.Studies have shown that nickel is a common cause of recurrent in-stent restenosis.

Objective:To study the impacts of pre-pregnancy body mass index (BMI), gestational diabetes mellitus (GDM) and gestational weight gain (GWG) on perinatal outcomes and mode of delivery.Methods:From November 2016 to December 2017, single-pregnancy women in early pregnancy (<13 weeks) regularly checked-up at our hospital were enrolled in this prospective cohort study and followed up until delivery. They were assigned into four groups according to pre-pregnancy BMI: obese group (≥28.0 kg/m 2), overweight group(24.0-<28.0 kg/m 2), normal group (18.5-<24.0 kg/m 2) and underweight group(<18.5 kg/m 2). A 75-g oral glucose tolerance test was performed at 24-28 weeks of pregnancy to screen for GDM. The optimal GWG was 11.0-16.0 kg for underweight group, 8.0-14.0 kg for normal group, 7.0-11.0 kg for overweight group and 5.0-9.0 kg for obesity group. The effects of pre-pregnancy BMI, GDM and GWG on perinatal outcomes and delivery mode were evaluated using multivariate logistic regression methods. Results:A total of 802 pregnant women were included. The incidences of pre-pregnancy overweight and obesity were 21.8% and 8.9%, respectively. The incidence of GDM was 14.1%. 57.2% of the participants experienced excessive GWG. The incidences of macrosomia, low birth weight and premature birth were 7.1%, 2.7% and 2.2%, respectively. The incidence of Cesarean delivery (C-section) was 37.7%. Pre-pregnancy obesity [adjusted odds ratio ( AOR)=4.355, 95% confidence interval ( CI) 1.900-9.980] and excessive GWG ( AOR=3.799, 95% CI 1.796-8.034) were independent risk factors for macrosomia. Excessive GWG was a protective factor for low birth weight ( AOR=0.279, 95% CI 0.084-0.928) and inadequate GWG was a risk factor for low birth weight ( AOR=10.954, 95% CI 3.594-33.382) and premature birth ( AOR=8.796, 95% CI 2.628-29.438). Compared with the normal group, overweight group had an increased risk of C-section ( AOR=1.817, 95% CI 1.119-2.949). Compared with pregnant women without pre-pregnancy overweight/obesity, GDM nor excessive GWG, any combination of two of the above-mentioned three factors increased the risks of macrosomia ( AOR=3.908, 95% CI 1.630-9.370) and C-section ( AOR=2.269, 95% CI 1.325-3.886). The risks of macrosomia and C-section were the highest when all three factors existed. Conclusions:Pre-pregnancy obesity and excessive GWG are independent risk factors for macrosomia and pre-pregnancy overweight is a risk factor of C-section. Exposure to any two of the three factors (pre-pregnancy overweight/obesity, GDM and excessive GWG) increases risks of macrosomia and C-section and the highest risk is observed when all three factors are present.

OBJECTIVETo understand the prevalence and spatial distribution of vitamin A deficiency (VAD) in China among children at ages under six years.

METHODSAbout 8,600 children under 6 years of age in 14 cities and 28 counties of 14 provinces were selected with stratified cluster sampling for survey, including interview with questionnaire for their family information and nutritional status. Blood specimens were collected for measuring serum level of VA with fluorescent spectrophotometry in laboratory.

RESULTSTotally, 8,669 children under 6 (2,877 in urban area and 5,792 in rural area) were surveyed in 14 provinces, with 4,629 males and 4,040 females. Eight cases of night blindness and seven cases of xerophthalmia were found among the children at ages of two to five years. Sixty-one mothers of the children in this group were also found suffering from night blindness. All the cases of night blindness and xerophthalmia both in children and mothers were living in rural areas. Based on their serum levels of VA, 11.7% of the all 1 018 children were diagnosed as VAD, with serum VA concentrations below or equal to 0.70 micro mol/L. Prevalence of VAD was 15.0% and 5.8% in rural (23.3% in the poverty-stricken counties) and urban areas, respectively, and 5.8%, 11.5% and 16.8% in the coastal, inland and remote areas, respectively. The average serum level of VA was 1.20 micro mol/L and 0.99 micro mol/L for urban and rural areas, respectively, with a national average of 1.06 micro mol/L. And, babies under six months of age with an average serum levels of VA < or = 0.70 micro mol/L accounted for 33.4%, and those at ages of four to five years with the same level of VA accounted for 8%. There was significant difference in serum levels of VA between ages, but no significant difference between genders.

CONCLUSIONVAD did exist in children of China, especially in the remote and poverty-stricken rural areas and VA supplementation is urgently needed for the children in these regions.

Child ; Child, Preschool ; China ; epidemiology ; Dietary Supplements ; Female ; Health Surveys ; Humans ; Infant ; Male ; Night Blindness ; complications ; Prevalence ; Rural Health ; statistics & numerical data ; Vitamin A ; administration & dosage ; therapeutic use ; Vitamin A Deficiency ; complications ; epidemiology ; prevention & control ; Xerophthalmia ; complications