[Effects of prenatal tetrandrine treatment on the expression of Kruppel-like factor 5 and Survivin in the lung of congenital diaphragmatic hernia rat model induced by nitrofen].

Objective: To investigate the effect of prenatal administration of tetrandrine on the expression of Kruppel-like factors 5 (KLF5) and Survivin in the lung of congenital diaphragmatic hernia (CDH) rat model. Methods: This experiment took simple random group method, 11 pregnant SD rats were randomly divided into three groups: normal control group (group NC, 3), model control group (group MC, 4), model+ tetrandrine group (group MT, 4). The rats in group MC and MT were given gavage administration with 125 mg nitrofen to induce CDH and the rats in group MT were administered with 30 mg/kg tetrandrine for three days (gestational age of 18.5, 19.5 and 20.5 d). All fetuses were delivered by caesarian section at the gestational age of 21.5 d to observe the formation of diaphragmatic hernia in 3 groups and to record the ratio of lung weight and body weight (Lw/Bw). The maturity situation of fetal lung tissue was evaluated by hematoxylin-eosin staining (HE staining). The expression of KLF5 and Survivin were detected by immune-histochemical staining and quantitatively analyzed by Western blotting methods.Single factor analysis of variance was taken among 3 groups to compare means of variables and the chi-square test was used to compare the difference of birth defect rates. Results: There was significant difference in CDH incidence between group MC [56.1%(37/66)] and MT[50.0%(28/56)](χ(2)=122.00, P<0.05). There was no significant difference in Lw/Bw among the three groups (F=0.985, P=0.376). The HE staining showed that pulmonary vascular-related and pulmonary alveolar-related developmental indexes in group MC were lower than those in the group NC, and those in the group MT were between the two groups.In addition, immunohistochemical staining showed that the average optical density (AOD) of KLF5 in group NC, MC, MT was 0.194 8±0.007 4, 0.212 1±0.004 9 and 0.192 7±0.001 9, respectively (F=14.53, P=0.002); and AOD of Survivin was 0.185 2±0.008 7, 0.209 2±0.003 6 and 0.192 8±0.007 5, respectively (F=12.31, P=0.003). The expression level of Survivin was positively correlated with that of KLF5 (r=0.993, P=0.039). Western blotting revealed that there were remarkable difference in the protein expression of KLF5 and Survivin among the three groups (F=4.29, 10.13, both P<0.05). Conclousions: Prenatal gavage of tetrandrine to CDH rats induced by nitrofen promotes the development of fetal lung parenchyma and vascellum, and reduces the expression level of KLF5 and Survivin protein in fetal lung, which may be the mechanism of tetrandrine in improving lung hypoplasia of CDH.

HBsAg may reappear following reactivation in individuals with spontaneous HBsAg seroclearance 8 years previously.

HBsAg reappearance may constitute not only a risk for liver disease but also an infectious source. We aimed to determine whether HBsAg may reappear after spontaneous HBsAg seroclearance. A cohort of 2999 HBsAg-positive subjects aged 30-55 years was recruited in Guangxi, China in 2004. HBsAg was tested every 6 months from July 2004 to June 2007, then, one more time in December 2013. The results showed that spontaneous HBsAg seroclearance occurred in 41 subjects in the first 3 years, giving a 0·54% annual seroclearance rate. Thirteen of the 41 subjects were randomly tested for HBsAg in 2013. Four subjects became HBsAg positive. S gene sequences of HBV were analysed from serum collected before seroclearance and after reappearance, respectively, for subject QS840 (11 and 12 clones), subject TN98 (13 and 13 clones) and subject WX227 (10 and 8 clones). Serotype, subgenotype and amino-acid substitution pattern in each sample collected after reappearance was observed in the sample collected before HBsAg seroclearance. Nucleotide similarity between the two sequences from each subject was >99% and five sequences from subject TN98 were the same. In conclusion, following reactivation, HBsAg may reappear in individuals with spontaneous HBsAg seroclearance many years previously.