Objective:To assess the effectiveness and feasibility of carrier detection for Spinal muscular atrophy (SMA) by using digital PCR assay.Methods:Peripheral blood samples were collected from 214 pregnant women who were routinely screened for SMA carriers, of which 204 were randomly selected samples and 10 were samples with known copy numbers of SMN1 exons 7 and 8. Samples with known copy numbers of SMN1 exons 7 and 8 were randomly mixed into the experiment to validate the performance of the digital PCR assay. The copy numbers of SMN1 exons 7 and 8 and SMN2 exons 7 and 8 in peripheral blood samples were detected by digital PCR assay. The results of SMN1 exons 7 and 8 were compared with those of the quantitative PCR method to assess the reliability and clinical performance of the digital PCR assay. Results:Among the 204 random samples, digital PCR has detected five samples with simultaneous heterozygous deletion of SMN1 exons 7 and 8, three samples with heterozygous deletion of SMN1 exon 8 only, and 196 samples with no deletion of SMN1 exons 7 and 8. Ten samples with known SMN1 exons 7 and 8 copy numbers were detected with the expected values. The digital PCR test results were fully consistent with that of the quantitative PCR. Conclusion:The results of digital PCR for the detection of copy number variation of SMN1 exons 7 and 8 were consistent with qPCR. Digital PCR assay was able to clearly distinguish the copy number of the target genes, therefore can be used for SMA carrier screening. Moreover, it can also detect copy number of SMN2 exons 7 and 8, which can provide more information for genetic counseling.

In order to provide basic information for the utilization and development of famous classical formulas containing Bletillae Rhizoma， this article systematically analyzes the historical evolution of the name， origin， harvesting and processing of Bletillae Rhizoma by reviewing the ancient materia medica， prescription books， medical books and modern literature. The research results showed that Baiji（白及） was the main name， some scholars took Baiji（白芨） as its main name， and there were many other names such as Baiji（白给）， Baigen（白根）， Baiji（白苙）. The mainstream source of Bletillae Rhizoma was the tubers of Bletilla striata， and drying， large， white， solid， root-free and skin removed completely were the good quality standards. With the promotion of wild to cultivated medicinal materials， there were certain differences between their traits， and the quality evaluation indexes should be adjusted accordingly. The origin of records in the past dynasties was widely distributed， with Guizhou and Sichuan having high production and good quality in modern times. The harvesting period is mostly in spring and autumn， and harvested in autumn was better. The processing and processing technology is relatively simple， and it was used fresh or powdered in past dynasties， while it is mainly sliced for raw use in modern times. Based on the results， it is suggested that the tubers of Bletilla striata of Orchidaceae should be used in the famous classical formulas， and it should be uniformly written as Baiji（白及）. And if the original formula indicates the requirement of processing， it should be operated according to the requirement， if the requirement of processing is not indicated， it can be used in raw form as medicine.

As a prenatal testing for chromosomal abnormalities, non-invasive prenatal testing (NIPT) has been integrated into prenatal healthcare service. NIPT has shown a high sensitivity and specificity for screening fetal trisomies 13, 18 and 21, and has attained excellent clinical results. With the propagation of the NIPT screening, international organizations have issued guidelines and comments for its clinical utility with regular updating. China has also developed guidelines for NIPT in 2016. NIPT guidelines in various countries have provided valuable guidance for its target diseases and suitable patient groups, but there has been few research data on its clinical application for special groups of patients. Based on the guidelines and comments of various professional bodies and published data on the clinical utility of NIPT, in addition with consideration of the conditions in China, clinical utility of NIPT for particular groups of pregnant women, including those with advanced maternal age, obesity, twin pregnancy and fetal ultrasonographic anomalies, are reviewed. The value of genetic counseling for NIPT is also emphasized, which is critical for the clinical application of NIPT.
China ; Chromosome Aberrations ; Female ; Humans ; Pregnancy ; Pregnant Women ; Prenatal Diagnosis ; Trisomy 13 Syndrome

OBJECTIVE：To provide reference for improving safe drug use in pregnant women. METHODS ：Retrieved from as domestic and foreign literature retrieval platforms as CNKI and Web of Science ，the literatures about drug use in pregnancy period were collected and analyzed by using the literature analysis function provided by the retrieval platform and CiteSpace 5.5R2 software. The collected data of annual volume ，author，organization，keyword co-occurrence and literature co-citation analysis of domestic and foreign literatures are visualized ，and the research focus was analyzed. RESULTS ：A total of 38 946 foreign literatures and 698 domestic literatures were included ，and domestic literatures were less than foreign literatures ；the literature volume on pregnancy drug use were increasing year by year at abroad，but the volume of domestic literatures fluctuates greatly and increased slowly. Foreign institutions and scholars worked closely together ，and built a close cooperation network ，especially in the United States. However ，the author team of domestic research was relatively scattered as a whole ，and the cooperation between researchers was less ，so a large cooperation team had not yet been formed. The most frequently cited literatures in the world were randomized controlled studies and large-scale cohort studies，the keyword “Risk”appears most frequently ，indicating that it focuses on the effects of pregnancy medication on pregnancy outcomes and offspring. The effects of antiepileptic drugs ，antidepressants and opioids used in pregnancy on pregnancy outcomes and child development were currently a research hotspot abroad. However ，there was a lack of large-scale sample and high-quality clinical research in China ，high-frequently keywords in domestic literatures indude “fetus”“hypertension in pregnancy ”etc.，which focused on the treatment of combined diseases in pregnancy and paid little attention to the long-term effects of medication in pregnancy. CONCLUSIONS：More and more attention had been paid to the study of drug use in pregnancy period. Domestic research needs to seize the international cooperation communication ，learns from the excellent experience of foreign research team ，and establishes domestic specific pregnancy medication research team. Relevant departments should introduce measures as soon as possible ，support medical institutions and drug production and developmententerprises to do a good job in the registration of pregnancy medication，establish the database of pregnancy medication inChina as soon as possible ，and improve the medication safety of pregnant patients.

Objective To observe the protective mechanism of angiotensin-converting enzyme (ACE) 2 on lipopolysaccharide (LPS)-induced hepatocyte inflammation by inhibiting P38 mitogen activated protein kinase (MAPK)/activator protein (AP)-1 pathway.Methods Rat liver BRL cells after immortalized culture were randomly divided into five kinds of groups:control group,LPS (10 μg/ mL) group,LPS + recombinant(r) ACE2 (5,10,20 ng/mL rACE2 for 30 min before cells stimulated with LPS) groups,LPS+ACE2 inhibitor MLN-4760 (10-7,10-6,105 mmol/L MLN-4760 for 30 min before cells stimulated with LPS) groups,LPS + rACE2 (20 ng/mL rACE2 for 30 min before cells stimulated with LPS) + P38MAPK inhibitor SB203580 (10-5 mmol/L SB203580 for 30 min before cells stimulated with LPS) groups.The changes in protein levels of ACE2,P38MAPK,p-P38MAPK and AP-1 were detected by western blot after LPS exposure for 6,12 and 24 hours,and the mRNA expressions of P38MAPK,AP-1 and tumor necrosis factor-α were quantified by real-time RT-PCR.Results Compared with control group,the protein levels of ACE2,P38MAPK and AP-1 were up-regulated in LPS-induced hepatic cells in a time-dependent manner,peaking at 12 h after LPS stimulation (all P＜0.05).Compared with LPS group,the mRNA expressions ofAP-l,P38MAPK,p-P38MAPK and tumor necrosis factor-oα decreased significantly in rACE2 group (all P＜0.05).The dose of 20 ng/mL rACE2 had the best inhibitory effects on the mRNA expression of AP-1 (0.12±0.002 vs.0.04±0.005,P＜0.01),P38MAPK (0.17±0.02 vs.0.02±0.002,P＜0.01) and p-P38MAPK(0.29±0.01 vs.0.02±0.01,P＜0.01)compared with LPS group.The mRNA expressions of AP-1,P38MAPK and p-P38MAPK increased in MLN-4760 group in a concentration dependent manner (all P＜0.05).Furthermore,the inhibitory effects of rACE2 on AP-1 and tumor necrosis factor-α levels were cancelled by SB203580.Conclusion The rACE2 can alleviate the LPS-induced hepatocyte injury by down regulating the P38MAPK/AP-1 signaling pathway.

Objective To prepare quercetin ( QT )-loaded polylactic-co-glycolic acid-D-α-tocopheryl polyethylene glycol 1000 succinate ( PLGA-TPGS) nanoparticles ( QPTN) and QT-loaded polylactic-co-glycolic acid ( PLGA) nanoparticles ( QPN) by using QT as model drug and PLGA-TPGS or PLGA as carrier materials, and to investigate the quality of the two nanoparticles. Methods QPTN and QPN were prepared by using the ultrasonic emulsification-solvent evaporation method, and their surface morphology,size and surface charge were detected by using a transmission electron microscope ( TEM) and a Nano ZS90 light scattering and laser Doppler anemometry, respectively. Drug loading ( DL) , entrapment efficiency ( EE) and in vitro drug release of QT in the two nanoparticles were determined by using a reverse phase-high performance liquid chromatography (RP-HPLC) on Hypersil C18 column (4.6 mm×250 mm, 5 μm) with methanol and 0.03% phosphoric acid (3︰2) as mobile phase, and the detective wavelength was 370 nm. Results TEM images exhibited that two nanoparticles were all spherical and regular. The average sizes of QPTN and QPN were (155.4±2.7) nm and (363.8±3.2) nm, while DL and EE of QPTN were approximately (21.6±2.8)%, (93.7±2.9)% (n=6), and DL and EE of QPN were approximately (15.0±1.5)%, (64.6± 1.6)% (n=6), respectively. Both of nanoparticles exhibited sustained release, and the cumulative QT release of QPTN and QPN reached (85.8±2.8)% and (68.6±1.4)% (n=6) at day 30, respectively, with a significant difference between them (P<0.05) . Conclusion QPTN gets smaller size, higher DL and EE, and exhibits sustained release, and the in vitro cumulative QT release is faster and more complete than QPN relatively.

OBJECTIVE:To study the in vitro uptake of Resibufogenin(RBG)lactic acid glycolic acid copolymer-water solu-ble vitamin E (PLGA-TPGS) in human liver cancer HepG2 cells,mouse ascites-type lymphatic metastasis of tumor HCa-F cells, and the toxicity on HepG2 cells. METHODS:RCPTN loading RBG and coumarin-6(C6)were prepared. Fluorescent inverted mi-croscope was used to observe the in vitro uptake by RCPTN HepG2,HCa-F cells. It was divided into negative control group,blank PLGA-TPGS nanoparticles(EPTN)group,5-fluorouracil solution(FS)group,RBG solution(RS)group,RBG/PLGA nanoparti-cles(RPN)group and RPTN group. WST-1 was conducted to investigate the optical density at 450 nm wavelength of HepG2 cells after 24,48,72 h incubated by FS,RS,RPN and RPTN with different final concentrations (1.25,2.5,5,10,20 μg/mL);the cell viability (CV) and half inhibitory concentration (IC50) were calculated. RESULTS:RCPTN distributed around the nucleus of HepG2,HCa-F cells. CV was decreased by RBG concentration increased in RPN group and RPTN group,and decreased by time prolonged;compared with FS group,CV in RPTN group was decreased(P<0.05 or P<0.01). IC50 of HepG2 cells incubated by FS,RS,RPN and RPTN was decreased by time prolonged,ordered by RS>FS>RPN>RPTN;IC50 incubated by RPN and RPTN for 48,72 h was obviously less than that of FS and RS(P<0.05 or P<0.01). CONCLUSIONS:RPTN can deliver RBG in-to HepG2,HCa-F cells,showing inhibition effect on HepG2 cells which is stronger than RPN,RS and FS.

Objective The aim of this study was to investigate the distribution and liver-targeting properties of quercetin (QT)/coumarin 6 (C6)-loaded polylactic-co-glycolic acid-D-α-tocopherol polyethylene glycol 1000 succinate (PLGA-TPGS) nanoparticles (QCPTN) in a hepatocarcinoma ectopic transplantation solid tumor model using HCa-F cell-bearing mice.Methods The QT concentrations in biological samples were determined using reverse phase-high performance liquid chromatography (RP-HPLC) analysis.After intravenous administration to mice in the QCPTN and QTS groups,the QT concentration in plasma and in different tissues was simultaneously analyzed at the different time points.Detection indexes (relative targeting efficiency,Re;targeting efficiency,Te) and fluorescence inversion microscopy images of the frozen tissue (liver,solid tumor,spleen,lungs,kidneys,and heart) slices were used for quantitatively and qualitatively evaluating the liver-targeting properties of QCPTN in solid tumor-bearing mice.Results Te of the QCPTN group in the plasma,liver,solid tumor,spleen,lungs,kidneys,and heart were all greater than 3,indicating that the area under the concentration-time curve (AUC) of liver was more than three times that of the plasma and other organs.Fluorescence inversion microscopy images showed that the green fluorescence of QCPTN was mostly observed in the liver.Conclusion Using HCa-F cell-beating mice,QCPTN was found to have better in vivo liver-targeting properties in hepatocarcinoma ectopic transplantation solid tumor.

Objective To investigate the therapeutic effects of Quercetin (QT)-loaded PLGA-TPGS nanoparticles (QPTN) on solid tumor-bearing mice with HCa-F hepatocarcinoma in vivo.Methods The model of HCa-F hepatocarcinoma solid tumor-bearing mice was established by implanting HCa-F cells into 48 mice.The mice were divided into 6 groups randomly:the negative control,empty PLGA-TPGS nanoparticles,5-Fluorouracil solutions (FS),Quercetin solutions (QTS),QT-loaded PLGA nanoparticles (QPN),and QPTN groups.Each group was treated using tail vein twice a day for 20 days;then,all mice were sacrificed.The increment tumor volumes and tumor growth inhibition rate were counted.Then,tumor specimens were prepared for hematoxylin & eosin (HE) staining and observed under a microscope.Results The results showed that the increment tumor volumes of mice in the QPTN,QPN,and FS groups were significantly smaller than that in the negative control group (P ＜ 0.05 or P ＜ 0.01).The tumor growth inhibition rate of the QPTN group was 59.07％,which was much higher than that of the QTS group (23.94％),the FS group (35.14％),and the QPN group (46.14％).The results of the HE staining on the tumor sections also indicated that the QPTN group showed a better therapeutic outcome compared to the other groups.Conclusion The QPTN has a better therapeutic effect on the model of solid tumor using HCa-F cells-bearing mice than the QPN,QTS,and FS.