The lncRNA PVT1/miR-590-5p/FSTL1 axis modulates the proliferation and migration of airway smooth muscle cells in asthma.

OBJECTIVE: Asthma is a prevalent chronic inflammatory airway disease that is characterised by airway remodelling and airway hyperresponsiveness. Abnormal proliferation and migration of airway smooth muscle cells (ASMCs) contribute to airway remodelling in asthma. However, the molecular mechanism underlying an increased ASMC mass in asthma remains elusive. Herein, we aimed at investigating the regulation of lncRNA PVT1 on ASMCs and focussing on the mechanism in the proliferation and migration. METHODS: Expression levels of lncRNA PVT1 and miR-590-5p in the serum collected from 24 children with asthma and 10 control children were determined by qRT-PCR. ASMCs proliferation and migration prior to and post platelet-derived growth factor subunit B (PDGF-BB) stimulation were examined by CCK-8 test and transwell assay. Dual-luciferase reporter assay was performed to determine miR-590-5p interaction with lncRNA PVT1 and follistatin-like 1 (FSTL1). Expression of lncRNA PVT1, miR-590-5p, FSTL1, C-Myc, cyclin D1, and cyclin-dependent kinase 1 (CDK1) was tested by quantitative real-time PCR (qRT-PCR) and immunoblotting analysis. RESULTS: The expression level of lncRNA PVT1 was higher but the expression level of miR-590-5p was lower in the serum of children with asthma than in control children. The expression level of lncRNA PVT1 was negatively correlated with the expression level of miR-590-5p in asthma. LncRNA PVT1 was upregulated upon PDGF-BB stimulation. LncRNA PVT1 knockdown by its specific shRNA repressed PDGF-BB-induced promotion of proliferation and migration in ASMCs and triggered an elevated miR-590-5p along with declined C-Myc, cyclin D1, and CDK1. The effects of lncRNA PVT1 knockdown on PDGF-BB-induced ASMCs were lost upon miR-590-5p inhibition. MiR-590-5p targeted FSTL1 gene and declined its expression, thus suppressing ASMC proliferation and migration following PDGF-BB stimulation and downregulating C-Myc, cyclin D1, and CDK1 expressions. The effects of miR-590-5p on PDGF-BB-induced ASMCs were lost upon FSTL1 overexpression. CONCLUSION: These results support the notion that the lncRNA PVT1/miR-590-5p/FSTL1 axis modulates ASMCs proliferation and migration following PDGF-BB stimulation, providing a potential therapeutic target to attenuate airway remodelling in asthma.

Prenatal care in combination with maternal educational level has a synergetic effect on the risk of neonatal low birth weight: new findings in a retrospective cohort study in Kunshan City, China.

OBJECTIVES: To investigate the dose-response relationship and synergetic effect of the maternal educational level and two measures of prenatal care on neonatal low birth weight (LBW) risk. METHODS: Data were derived from the Perinatal Health Care Surveillance System (PHCSS) from January 2001 to September 2009 in Kunshan City, Jiangsu province, eastern China, which included data on 31412 women with a normal birth weight delivery and 640 women with a LBW delivery. Logistic modelling was performed to estimate the association including the joint effects with odds ratio (OR) and 95% confidence interval (CI) between the prenatal care measures and LBW risk after adjusting for the potential confounders. The dose-response relationship between the number of prenatal care visits and the risk of LBW was investigated by modeling the quantitative exposure with restricted cubic splines (RCS). RESULTS: There was a significant synergetic effect on the LBW risk between maternal educational attainment and the number of prenatal care visits (χ(2) = 4.98, P = 0.0257), whereas no significant maternal educational attainment interaction was found with the week of initiation of prenatal care after adjusting for relevant confounding factors (χ(2) = 2.04, P = 0.1530), and the LBW risk displayed a 'U-shape' curve tendency among the different number of prenatal care visits (P for nonlinearity = 0.0002) using RCS. In particular, the ORs were approaching the curve's bottom when the women had 9 or 10 prenatal care visits. Comparing with 5 prenatal care visits, the ORs and 95%CI of LBW risk for 7, 9, 11 and ≥ 13 visits were 0.92 (0.82-1.03), 0.50 (0.38-0.66), 0.62 (0.47-0.82), and 0.99 (0.61-1.60), respectively. CONCLUSIONS: Our findings suggest that appropriate prenatal care, in combination with a higher maternal educational level, can produce a protective interaction effect on LBW risk. Reasonable health resource assignment for different social statuses should be taken into account by policy-makers in developing countries.

[Optimal extraction of puerarin from Pueraria lobata based on central composite design and response surface methodology].

OBJECTIVE: To optimize the parameters of extraction technology for Puerarin from Pueraria lobata by Central Composite Design-Response Surface Methodology. METHODS: Used the ratio of dosage liquid and extraction time as the examination factor,the content of puerarin for the extraction percentage as the index. The multielement linear model and the polynomial model were used to describe the mathematics relation with response surface. RESULTS: The optimal extraction technology was as follows: ratio of herb-to-solvant was 1: 8.88 (W/V) and extraction time was 150 min. The coefficient correlation was 0.9977 and closed to the predicted value. CONCLUSION: The results are accordant with model predictions. The technology can be used as the extraction process of puerarin from Pueraria lobata.

[Association between serum bisphenol-A and recurrent spontaneous abortion: a 1:2 case-control study, China].

OBJECTIVE: This study was to investigate the association between serum Bisphenol-A (BPA) and unexplained recurrent spontaneous abortion (RSA). METHODS: A hospital-based 1:2 matched case-control study was conducted.Sixty-two patients with unexplained recurrent abortion were included and matched with 2 normal controls by factors as age (± 2 years), living in the same district and the same gestational age.The levels of BPA in serum for 62 cases and 108 controls were detected under high performance liquid chromatography after fluorescent derivatization. Levels of serum BPA in each case was compared with that in control of age, BMI, education levels, occupation, exposure for passive smoking. RESULTS: The values of serum BPA in cases and controls were (0.009 ± 0.002) and (0.004 ± 0.012) µg/ml, respectively. The levels of serum BPA in cases was significantly higher than in controls (Z = 3.506, P = 0.0005). After adjusted by age, BMI, education levels, occupation, passive smoking history and other factors, when compared to BPA below 0.004 µg/ml. The adjusted ORs were 4.39 (1.15 - 16.71) for BPA levels between 0.004 µg/ml and 0.012 µg/ml, and 4.95 (1.77 - 13.82) for BPA over 0.012 µg/ml. The risk of unexplained recurrent spontaneous abortion increased progressively with the growth of serum BPA levels (χ(2) = 9.179, trend test P = 0.0024). There were significant differences on BPA among controls that with histories of two, three or more abortions (the levels were 0.004, 0.008, 0.018 µg/ml, respectively, F = 8.92, P = 0.0002). CONCLUSION: High BPA level might be associated with unexplained recurrent spontaneous abortion.

[Association between gestational weight gain and birth weight (2001-2009): a cohort study carried outin Kunshan city, Jiangsu province, China].

OBJECTIVE: To identify the association between gestational weight gain and birth weight over the past 9 years in Kunshan city, Jiangsu province, China. METHODS: This population-based study was conducted between 2001 to 2009. Data were retrieved from Perinatal Monitoring System of Maternal and Child Health Care Hospital of Kunshan. The study population consisted of 33 631 women and singleton live fetus. Gestational weight gain was defined as the total weight gain during the last and first prenatal care program and divided by the interval weeks. RESULTS: From 2001 to 2009, the average incidence of low birth weight was 1.86%, while the average incidence of macrosomia was a bit higher, fluctuating around 8.47%. On those underweight mothers, after adjustment for potential confounders, and stratified by the BMI levels, which were evaluated at the first prenatal care program, we found that weight gain in the 3rd and 4th intervals, could reduce the risk of low birth weight (less than 2500 g). With those mothers with normal-weight, weight gain in the 2 nd, 3 rd and 4th intervals, would reduce the risk of low birth weight. Risks in the 4th quantile among underweight and normal-weight group were prevalence odds radio (POR) 95%CI: 0.51 (0.32-0.80) and 0.58 (0.42-0.79), respectively. The risks showed a significant downward trend in underweight and normal-weight groups with increased gestational weight gain. As for macrosomia (≥4000 g), the risks increased (POR 95%CI) 4.69 (2.82-7.81) in underweight, 4.15 (3.43-5.03) in normal-weight, in overweight, 3.64 (2.62-5.06) and 1.96 (1.48-2.60) in obese mothers with increased levels of gestational weight gain. Trend tests indicated that the risks of marcosomia increased in all levels of BMI, with the increase of gestational weight gain. CONCLUSION: Findings from this population-based study suggested that gestational weight gain could reduce the risks of low birth weight among underweight and normal-weight groups, while increase the risks of macrosomia in all parturients, as compared with lowest levels of gestational weight gain.

[A correlative study on Bisphenol A and recurrent spontaneous abortion].

OBJECTIVE: This study was to investigate the association of Bisphenol A and unexplained recurrent spontaneous abortion. METHODS: A hospital-based 1:1 matched case-control study was conducted. Sixty patients with unexplained recurrent abortion were included. Each case was matched with one normal control by age (± 2 years), living district and the same gestational age. The levels of Bisphenol A in urine for 60 cases and 60 controls were detected using high performance liquid chromatography after fluorescent derivatization. The levels of urinary Bisphenol A in case was compared with that in control in education levels, occupation, smoking history. Data was analyzed by means of Wilcoxon-test, Student-Newman-Keuls after rank transform, univariate and multivariate conditional Logistic regression analysis. The software used was SAS 9.1.3. RESULTS: The values of urinary Bisphenol A in cases and controls were (0.10 ± 0.21) µg/ml, (0.03 ± 0.08) µg/ml, respectively. The level of urinary Bisphenol A in cases was significantly higher than that in controls (Z = 3.988, P < 0.0001). The urinary Bisphenol A levels in cases were significant higher than those in controls from senior middle school (the levels were 0.10, 0.06 µg/ml respectively, Z = 1.996, P = 0.0459), college (the levels were 0.14, 0.03 µg/ml respectively, Z = 2.586, P = 0.0097), workers or farmers (the levels were 0.08, 0.03 µg/ml respectively, Z = 2.265, P = 0.0235), businessmen (the levels were 0.10, 0.03 µg/ml respectively, Z = 2.544, P = 0.0109), and no passive smokers (the levels were 0.09, 0.03 µg/ml respectively, Z = 3.767, P = 0.0002). After adjustment by age, body mass index, marital status during pregnancy and other factors, compared to Bisphenol A below 0.06 µg/ml, the adjusted OR was 4.03 (1.67 - 9.74) for Bisphenol A levels between 0.06 µg/ml and 0.20 µg/ml, and was 5.46 (1.95 - 15.27) for Bisphenol A over 0.20 µg/ml. The risk of unexplained recurrent spontaneous abortion increased progressively with the growth of urinary Bisphenol A levels (χ(2) = 13.042, trend test P = 0.0003). There were significant differences on Bisphenol A among controls, two abortions, and three or more abortions (the levels were 0.03 µg/ml, 0.09 µg/ml, 0.21 µg/ml respectively, F = 9.04, P = 0.0002). CONCLUSION: Exposure to Bisphenol A may be associated with unexplained recurrent spontaneous abortion.

[An investigation of Gram-positive bacteria infection in an intensive care unit during 6 years].

OBJECTIVE: To retrospectively investigate the risk factors, distribution, antibiotic resistance of infection with Gram-positive (G+) bacteria in an intensive care unit (ICU), so as to provide the reference for clinical prevention and treatment. METHODS: A retrospective analysis of clinical data of 83 patients with G+ bacteria infection in ICU from January 2003 to December 2008 was done. RESULTS: Of the 125 strains of G+ bacteria from 83 patients, Staphylococcus was the main organism (63.2%, 79/125). The prognosis of the patient was related with surgical operation (chi2=9.107, P=0.003), gastric intubation (chi2=4.053, P=0.044), complication (chi2 5.908, P=0.015) and the use of immunosuppressant (chi2=5.761, P=0.016). Multi-bacterial infection was related with surgery (chi2=8.847, P=0.003) and tracheostomy (chi2=10.445, P=0.001). The antibiotic susceptibility test in vitro showed that G+ bacteria displayed multi-resistance to antibiotics, but all of G+ bacteria were sensitive to vancomycin (resistance rate was 0). CONCLUSION: Staphylococcus was the most common pathogen of G+ bacterial infection in ICU. Further surveillance of bacterial resistance is warranted in ICU, and antimicrobial drugs should be used according to the result of susceptibility test. Taking account of the antibiotic resistance and risk factors of G+ bacteria infection in ICU, the infection could be controlled and the death rate could be cut down when appropriate measures are taken.

TKI-31 inhibits angiogenesis by combined suppression signaling pathway of VEGFR2 and PDGFRbeta.

Tyrosine kinases have been strongly implicated as therapeutic targets that influence the angiogenic process in growing tumors. In this study, we revealed that TKI-31 is a potent broad spectrum tyrosine kinase inhibitor, which inhibits vascular endothelial growth factor receptor 2 (VEGFR2), platelet-derived growth factor receptor beta (PDGFRbeta) and also inhibits kinases of other class, such as c-Kit and c-Src on molecular base, but showed no activity against vascular endothelial growth factor receptor 1 (VEGFR1) and epidermal growth factor receptor (EGFR). TKI-31 inhibits VEGF-induced phosphorylation of VEGFR2 in endothelial cells as well as PDGF(BB)-induced phosphorylation in fibroblast cells, and leading to the inhibition of down-stream signaling triggered by these receptors such as PI3K/Akt/mTOR, MAPK42/44(ERK) and paxillin. TKI-31 also inhibited VEGF-induced endothelial cells proliferation, migration and their differentiation into capillary-like tube formation. Its anti-angiogenic property was further confirmed by the inhibition of neovascularization on CAM, in vivo. These results collectively highlight the therapeutic potential of this compound for the treatment of solid tumors and other diseases where angiogenesis plays an important role.

[Trend of the CD23+ B cells in children with infectious mononucleosis caused by Epstein-Barr virus].

OBJECTIVE: Epstein-Barr virus (EBV) is a common causative agent of infectious mononucleosis (IM) and capable of efficiently immortalizing primary B cells into continuously growing lymphoblastoid cells in vitro. As B cell activation antigen, CD23 expression is induced by EBV infection of B cells and remains constitutively expressed at high levels in virtually all EBV-immortalized cells, which have been strongly linked to the development of B-cell lymphoproliferative disease and lymphoma. Whereas previous studies were performed in vivo in animals or ex vivo cultures, the present study aimed to explore the role of EBV-immortalized cells (CD23(+)/CD19(+)) in vivo analysis of children with EBV-IM. METHODS: In a prospective trial, a group of 30 patients with IM (18 boys and 12 girls) with mean age of 3.9 +/- 1.3 years (range 6 months to 8 years) were enrolled. Clinical diagnosis of IM was confirmed based on fever, lymphadenopathy, splenomegaly, lymphocytosis (> 50%), atypical lymphocytes (> 10%) in blood smears and the elevated levels of IgM antibody against EBV capsid antigen. The day of onset of fever was recognized as day 1 of illness. Blood samples taken during acute (3 - 5 days), early convalescent (about 11 - 15 days) and convalescent phase (about 30 - 45 days) were analyzed for expressions of CD19(+)/CD23(+), CD23, CD19 on peripheral blood mononuclear cells by flow cytometry (FCM) and was compared with those of control group. RESULTS: (1) The levels of CD23(+)/CD19(+) and CD23 expressions were markedly decreased in acute stage [CD23(+)/CD19(+) (2.22 +/- 1.47)%, (132 +/- 91)/mm(3); CD23 (3.12 +/- 1.88)%, (195 +/- 102)/mm(3)] and in early convalescent stage [CD23(+)/CD19(+) (4.51 +/- 2.25)%, (166 +/- 85)/mm(3); CD23 (5.55 +/- 2.76)%, (231 +/- 130)/mm(3)] in patients with IM as compared with those of the healthy controls [CD23(+)/CD19(+) (6.71 +/- 2.25)%, (215 +/- 68)/mm(3); CD23 (7.85 +/- 3.09)%, (249 +/- 86)/mm(3), respectively]. The earlier the history was, the lower the expressive levels were. The levels of CD23(+)/CD19(+) expressions returned to, but those of CD23 expressions exceeded, normal level in convalescent stage [CD23(+)/CD19(+) (6.72 +/- 2.16)%, (213 +/- 108)/mm(3); CD23 (9.46 +/- 2.73)%, (366 +/- 200)/mm(3)]. (2) There was a positive correlation in the expressions of CD23(+)/CD19(+) and CD23 among the three stages (P < 0.01). The positive correlation between the expressions of CD23(+)/CD19(+) and CD19 only occurred during acute stage (P < 0.01). There was no correlation between the expressions of CD23 and CD19 (P > 0.05). CONCLUSION: EBV-immortalized cells and CD23(+) cells were inhibited effectively during the acute and early convalescent stage of IM. With the recovery of the disease, they gradually recovered and the levels of CD23 expressions exceeded normal level in convalescent stage.

[The inhibition pathway of the EBV-immortalized cells in children with infectious mononucleosis].

OBJECTIVE: To explore the inhibition pathway of the EBV-immortalized cells (CD23(+)) in children with infectious mononucleosis (IM) caused by Epstein-Barr virus. METHODS: The expressions of CD23, CD19, CD95, Bcl-2 and the co-expressions of CD23CD95, CD19CD23 on peripheral blood mononuclear cell (PBMC) were analyzed by flow cytometry (FCM) during acute phase, early convalescent phase and convalescent phase of 34 EBV-IM children and compared with that of 24 healthy donors. RESULTS: (1) The levels of CD23(+) and CD23(+)CD19(+) cells decreased and CD95(+), CD95(+)CD23(+), Bcl-2(+) cells increased markedly in IM patients in acute phase [CD95(+) cells (19.43 +/- 8.46)%; CD95(+)CD23(+) cells (1.81 +/- 1.71)%; Bcl-2(+) cells (23.41 +/- 26.47)%] and early convalescent phase [CD95(+) cells (12.94 +/- 5.05)%; CD95(+)CD23(+) (1.05 +/- 1.20)%; Bcl-2(+) cells (10.54 +/- 9.68)%], as compared with those of healthy controls [CD95(+) cells (10.39 +/- 2.90)%; CD95(+)CD23(+) cells (0.50 +/- 0.46)%; Bcl-2(+) cells (7.25 +/- 2.88)%]. The earlier the course of IM, the more abnormal the expressive levels. All the abnormal results returned to normal in convalescent phase. (2) Positive relationships were observed between the expressions of CD95(+)CD23(+) cells and that of CD23(+) cells, CD23(+)CD19(+) cells during acute and early convalescent phase, the expressions of Bcl-2(+), CD3(+) cells and CD23(+), CD23(+)CD19(+) cells during acute phase, the expressions of CD95(+)CD23(+) cells and Bcl-2(+) cells during acute phase, and the expressions of CD95(+)CD23(+) cells and CD95(+) cells during convalescent phase. CONCLUSION: The results indicate that CD95L-CD95 mediated apoptosis plays an important role in eliminating EBV-immortalized cells, which is counteracted partly by Bcl-2.

[Role of the B lymphocytes in children with infectious mononucleosis caused by Epstein-Barr Virus].

OBJECTIVE: Infectious mononucleosis (IM) is a lymphoproliferative disease caused primarily by the Epstein-Barr virus (EBV) infection. The initial viral infection by EBV occurs in B lymphocytes and is followed by an extensive proliferation of T lymphocytes. Previous studies on immunity to EBV (including IM) have mainly focused on activation of peripheral blood T cells, which are responsible for the lymphocytosis in blood during acute IM. B cells, regarding CD23 as their activation marker, are the target cells of EBV infection. There are few reports on their effect in patients with IM. The role of them during acute IM is not known yet. The present study aimed to explore the action of B cells in patients with IM. METHODS: In a prospective trial, a group of subjects comprised 22 patients with IM (14 boys and 8 girls) with mean age of 3.48 +/- 0.81 years (range 7 months to 8 years). Clinical diagnosis of IM was confirmed based on fever, lymphadenopathy, splenomegaly, lymphocytosis (> 50%), atypical lymphocytes (> 10%) in blood smears and the elevated levels of IgM antibody against EBV capsid antigen. The day of onset of fever was recognized as day 1 of illness. Blood samples taken during acute (3 - 5 days) and convalescent phase (about 15 days) were analyzed for expressions of CD19, CD19(+)/CD23(+) on PBMC by flow cytometry (FCM) and was compared with those of control group. The number of the days with fever was recorded. RESULTS: (1) The levels of CD19 and CD19(+)/CD23(+) expressions were markedly decreased in acute stage [CD19 (5.63 +/- 2.91)%, (387 +/- 178)/mm(3), CD19(+)/CD23(+) (2.45 +/- 1.87)%, (160 +/- 99)/mm(3)] and in convalescent stage [CD19 (12.49 +/- 5.70)%, (428 +/- 156)/mm(3), CD19(+)/CD23(+) (5.05 +/- 2.79)%, (172 +/- 78)/mm(3)] in patients with IM as compared with those of the healthy controls [CD19 (16.20 +/- 2.80)%, (545 +/- 150)/mm(3); CD19(+)/CD23(+) (7.08 +/- 2.78)%, (249 +/- 136)/mm(3)]. The earlier the specimens were taken after onset, the lower the expressed levels were. (2) There was a positive correlation of the expressions of CD19 and CD19(+)/CD23(+) between acute and convalescent stage (P < 0.01);there was also a positive correlation between the expressions of CD19 and CD19(+)/CD23(+) during acute and convalescent stage (P < 0.01). (3) A negative correlation was found between the duration of fever and the level of CD19 and CD19(+)/CD23(+) in acute stage (P < 0.01). CONCLUSION: The results indicate that B cells and CD23(+) B cells were significantly inhibited during the onset of IM in the patients, that with the recovery of the disease, the condition was gradually improved, and that the more evidently the CD19 and CD19(+)/CD23(+) decreased, the more serious the clinical symptoms were and the longer time the recovery needed. The levels of CD19 and CD19(+)/CD23(+) expressions may be useful in diagnosis and predicting the severity.