ABSTRACT
Objective To study the influences of Yang-warming, Blood-activating, and Toxin-removing Recipe(YBTR)-containing serum sampled under different conditions on the proliferation of human umbilical vein endothelial cells(HUVECs). Methods HUVECs were cultured in vitro for the experiment. Pharmacological serum for the experiment was prepared as follows: Thirty-two SD rats (male in half) were randomly divided into 4 groups, namely normal saline group and low-, middle-, and high-dose YBTR groups (at the intragastric dosage of 10.35, 31.05, 93.15 g·kg-1 respectively, twice a day). The pharmacological serum was taken from one female rat and one male rat in various groups at 4 time points, i.e. at hour 1, 2 after first intragastric administration on the fourth feeding day, and at hour 1, 2 after first intragastric administration on the sixth feeding day(abbreviated as D3H1, D3H2, D5H1, D5H2 respectively). The effects of YBTR-containing serum on the proliferation of HUVECs were observed by CCK-8 assay method. Results The difference of proliferation-inhibition rate of HUVECs was statistically significant after treated with YBTR-containing serum prepared from rats of different genders at different time(Pgender=0.000<0.01, Ptime=0.000<0.01). The difference of interaction of time and gender was also significant (Ptime × gender=0.001<0.01), and the effect at D3H1 and D5H1 varied with the gender (PD3H2×gender = 0.000 < 0.01, PD5H1×gender = 0.002 < 0.01). The inhibitory action of YBTR-containing serum became stronger with the increase of the dosage of serum of female rat at D3H2, and the inter-group difference was statistically significant (P < 0.05), the effect showing concentration-dependent tendency. The inhibition of HUVECs proliferation reached the peak after treated by various doses of YBTR-containing serum from the female rat at D3H2, while the inhibition arrived to the peak after treated by low- and middle-dose YBTR-containing serum from the male rat at D5H1, and the inhibition arrived to the peak in the group of high-dose YBTR-containing serum from the male rat at D3H1. Conclusion The inhibitory action of YBTR-containing serum on the proliferation of HUVECs was stronger when the serum was taken from the female rat at D3H2.
ABSTRACT
Objective To carry out mutation analysis of deafness-associated genes for deaf newborns and their parents, and to estimate the recurrence risk for their parents to have deaf descendants.Methods Suspected cases of inherited deafness were identified by neonatal hearing screening and questionnaires. Genomic DNAs of suspected cases and their parents were extracted from their peripheral blood samples . Common deafness-associated genes(i.e. GJB2,SLC26A4 and 12S rRNA genes)were amplified by polymerase chain reaction(PCR),and those PCR products were sequenced for the mutation analysis.Results From 2013 to 2016, 193 cases of deafness were found in neonatal hearing screening,29 cases of suspected as hereditary deafness were screened,and 17 out of 29 cases were found to have mutations in deafness-associated genes(detection rate:58.62%). GJB2 homozygous mutations were identified in two cases and their parents,and the recurrence risk to have deaf descendants was 100%. Four cases of suspected hereditary deafness had GJB2 homozygous mutations,and their parents were both GJB2 mutation carriers. There was one case with SLC26A4 homozygous mutations,and their parents were both SLC26A4 mutation carrier. Two cases were detected to have GJB2 V371 homozygous mutations,and their parents were both GJB2 V371 mutation carriers. For those seven parents carrying deafness-associated mutations above,the recurrence risk of deafness for their descendants was 25%.Conclusion In addition to hearing screening,the genetic diagnosis of deafness-associated genes is helpful to clarify the cause of suspected neonatal hereditary deafness,and can provide objective reproductive counseling and guidance for those deaf parents or parents with deaf children.