OBJECTIVE:To establish a method for the simultaneous determination of chlorogenic acid,evodiamine,rutecar-pine in different places of Evodia rutaecarpa. METHODS:UPLC was performed on the column of ACQUITY UPLC BEH C18 with mobile phase of acetonitrile-0.1% Phosphoric acid aqueous solution(gradient elution)at a flow rate of 0.40 ml/min,the detection wavelength was 326 nm and 220 nm,column temperature was 30 ℃,and the volume injection was 2 μl. RESULTS:The linear range was 7.67-76.67 μg/ml for chlorogenic acid(r=0.999 2),13.33-133.33 μg/ml for evodiamine(r=0.999 7)and 13.33-133.33μg/ml for rutecarpine(r=0.999 8);the limits of quantitation were 0.11 ng,0.05 ng and 0.05 ng,the limits of detection were 0.03 ng,0.01 ng and 0.01 ng,respectively;RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 96.19%-101.90%(RSD=2.19%,n=6),95.35%-101.16%(RSD=2.27%,n=6)and 95.92%-98.98%(RSD=1.33%,n=6),re-spectively. CONCLUSIONS:The method is rapid and accurate,and suitable for the simultaneous determination of chlorogenic ac-id,evodiamine,rutecarpine in different places of E. rutaecarpa.

Objective To raise ginsenosides yield from Panax ginseng by supercritical CO2 reverse microemulsion extraction.Methods Bis-(2-ethylhexyl)sodium sulphosuccinate(AOT)/ethanol/water/supercritical CO2 reverse microemulsion extraction was carried out to extract gingsenosides.Results The ginsenosides extracting rate by supercritical CO2 reverse microemulsion extraction was 3.2 times that of by ethanol/water/supercritical CO2 extraction in extracting pressure 25 MPa,extracting temperature 45 ℃,extracting time 4 h,and CO2 flow rate 2.0 L/h,respectively.The ginsenosides extracting yield increased with the increasing of the water amount and the extracting pressure,increased first and then decreased with the increasing of AOT concentration and the extracting temperature.The ginsenosides extracting yield with P.ginseng soaked with water before extraction was 1.3 times that with P.ginseng soaked with water during the extraction.Conclusion Combined the experimental results with the theoretic deduction,the mechanism of supercritical CO2 reverse microemulsion extraction is that the polar water pools of reverse microemulsion can make more ginsenosides dissolved.

Objective To up-regulate the expression of Glil gene in periodontal ligament stem cells ( PDLSCs) and to explore the effect of Glil gene on PDLSCs proliferation and osteogenesis differentiation by establishing Glil gene adenovirus vectors. Methods Subcloned Glil to viral backbone vector Adtrack-CMV and transfered the established vector to 293T cells, which was to acquire the virus particles. Trans-fected aim cells,namely PDLSCs,with these virus. Detected its effect on PDLSCs proliferation with CCK-8 assay, and detected the expression of Glil and the bone-related markers ALP and Runx2 through Western blot. Results An adenovirus vector, which were over expressed Glil gene, was successfully constructed and transfected to PDLSCs. Compared with the empty vector group and normal group, the over expressed one had a much slower proliferation rate in CCK-8 assay (P=0. 003). Western blot showed that ALP and Runx2 can be overexpressing os-teogenic differentiated after PDLSCs successfully transfected with the Glil gene. Conclusion Over expressing Glil gene would lead to a much slower proliferation rate in the PDLSCs and an increase of the bone-related markers. It is concluded that Glil can enhance the osteogenic dif-ferentiation capacity in PDLSCs.

Objective To investigate the effect of β-catenin on the osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) under mechanical tension.Methods PDLSCs were seperately cultured in vitro from the periodontium attached at freshly extracted teeth.β-catenin-targeting agonist or inhibitor was applied to the PDLSCs to upregulate or downregulate the expression of activity of β-catenin protein.The expression of β-catenin protein and the osteogenetic related markers(ALP,BMP2,Runx 2) under the mechanical tension with different period(0,6,12,24 h) were assessed with Western blot in the PDLSCs irritated with agonist or inhibitor.Results Compared with the PDLSCs without mechanical tension,the expression of osteogenesis related marker proteins,ALP and Runx 2,in PDLSCs were enhanced under the circular mechanial tensile stress (P ＜ 0.05).In the PDLSCs irritated with WAY-262611,the β-catenin-targeting agonist,the expression of osteogenetic related markers ALP in the PDLSCs was improved during the first 12 h period of mechnical tensile load,which was significantly higher than that in the DMSO group.Conclusion Wnt/β-catenin signaling pathway plays an important role in the early osteogenesis differentiation of PDLSCs under cyclic tension.The β-catenin promotes osteogenic differentiation of PDLSCs under mechanical tension.

Objective To analyze the clinical characteristics of structural malformations in one of monochorionic diamnionic twins (MCDA). Methods The clinical data of 77 MCDA patients with structural malformations diagnosed by ultrasound were retrospectively reviewed from January 2012 to May 2017. The distribution of structural malformations, prenatal chromosomal karyotyping and pregnancy outcomes were analyzed. Results (1)Among the 77 MCDA patients with structural malformations, the single malformation accounted for 79％(61/77), the multiple malformations accounted for 21％(16/77). And there were a total of 94 types of malformations, the top three malformations were neurological malformations (32％, 30/94), cardiovascular malformations (29％,27/94) and twin reversed arterial perfusion sequence (TRAPS;10％,9/94). (2)Among the 77 patients with structural malformation, 64 cases (83％, 64/77) were examined for fetal chromosomes, of whom 14 cases (22％,14/64) were examined for fetal chromosomes of both twins, with 1 case (1/14) of discordant fetal chromosome. (3)Among the 77 patients, 4 cases (5％, 4/77) with severe fetal malformations terminated pregnancy. Totally 29 cases (38％, 29/77) with severe malformations were treated with selective fetal reduction, among whom 7 cases (24％, 7/29) experienced unexplained fetal death within 24 hours after the operation;2 cases (7％, 2/29) happened inevitable abortion, and 2 cases (7％, 2/29) underwent unexplained fetal death during the late pregnancy. Of the remaining 44 patients (57％,44/77) with expectant treatment, 13 cases (30％, 13/44) occurred twin transfusion syndrome (Ⅱ-Ⅳ), and were treated with fetoscopic laser occlusion. Eight patients had 2 survival twins, 4 patients delivered 1 survival twin, and 1 patient had dead twins. Conclusions The most common malformations in MCDA twins are the nervous malformations, cardiovascular malformations and TRAPS. The chromosome karyotype of MCDA twins with structural malformations are sometimes discordant, and separate samling of the twins is suggested for prenatal diagnosis. Selective fetal reduction could be given to severe structural malformation in MCDA patients safely and effectively. For non-severe structural malformation in MCDA patients with twin transfusion syndrome, fetoscopic laser occlusion is safe and effective.

Objective To explore the effects of early enteral nutrition (EN) combined with probiotics on intestinal flora and immune function in patients with severe ischemic stroke. Methods Sixty-nine severe ischemic stroke patients were admitted and continuously enrolled in Taizhou First People's Hospital from June 2017 to June 2018, and they were randomly divided into an EN combined with probiotics group (35 cases) and a simple EN group (34 cases). Early EN support was given to both groups and probiotics (Live Combined Bifidobacterium, Lactobacillus and Enterococcus capsules) was added to the EN combined with probiotics group, 0.42 g each time, 3 times a day for 14 days. The changes of serum inflammatory markers [hypersensitive C-reactive protein (hs-CRP), procalcitonin (PCT), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10)], intestinal flora (Bifidobacterium, Lactobacillus, Clostridium, Enterobacter, Enterococcus, Bacteroides), intestinal mucosal barrier [endotoxin, D-lactic acid, diamine oxidase (DAO), intestinal fatty acid binding protein (I-FABP) ], and immune indexes [immunoglobulins (IgA, IgG, IgM), human leukocyte DR antigen (HLA-DR)] were observed in two groups of patients after treatment. Results With the prolongation of time, Bifidobacteria, Lactobacilli, HLA-DR and IgA, IgG, IgM after EN in both groups all decreased first and then had a tendency of increase, all reaching the lowest value on the EN 3rd day and then gradually elevated arriving at the peak value on the EN 14th day, and the levels in EN combined with probiotics group were significantly higher than those in the simple EN group [Bifidobacterium (×107 cfu/g): 8.31±1.49 vs. 7.49±1.32, Lactobacillus (×107 cfu/g): 8.04±1.45 vs. 7.19 ±1.37, HLA-DR: (67.22±9.11)% vs. (61.21±9.69)%, IgA (mg/L): 170.34±40.13 vs. 149.54±38.76, IgG (g/L):4.88±0.88 vs. 4.31±0.86, IgM (mg/L): 879.47±100.82 vs. 821.52±97.75, all P < 0.05]. With the prolongation of time, the Clostridium, Enterobacter, Enterococcus, Bacteroides, hs-CRP, PCT, TNF-α, endotoxin, D-lactic acid, DAO, I-FABP after En in both groups all increased first and then had a tendency of decrease, reaching the highest level on the EN 3rd day, then gradually decreased arriving at the valley value on the EN 14th day, and the levels in the EN combined with probiotics group were significantly lower than those in the simple EN group [Clostridium (×107 cfu/g): 5.23±0.87 vs. 5.79±0.91, Enterobacter (×107 cfu/g): 7.45±1.21 vs. 8.62±1.32, Enterococcus (×107 cfu/g): 7.32±1.05 vs. 8.12±1.23, Bacteroides (×107 cfu/g): 9.16±1.35 vs. 9.87±1.42, hs-CRP (mg/L): 18.45±12.98 vs. 25.47±15.55, PCT (ng/L): 3.24±1.21 vs. 4.18±1.32, TNF-α (ng/L): 9.43±8.69 vs. 13.59±9.45, IL-10 (μg/L): 39.45±10.72 vs. 48.52±11.42, endotoxin (U/L): 6.74±2.12 vs. 9.21±3.28, D-lactic acid (mg/L): 98.74±20.74 vs. 114.78±19.89, DAO (mg/L): 21.45±8.49 vs. 29.47±9.41, I-FABP (ng/L): 1.4±0.2 vs. 1.5±0.2, all P < 0.05]. Conclusion Early EN combined with probiotics can effectively regulate the intestinal flora and intestinal mucosal barrier function, reduce the level of inflammatory response and enhance the body immunity in patients with severe ischemic stroke.

Objective To understand the relationship between perinatal factors and congenital hypothyroidism(CH)and provide scientific evidence for the prevention of CH. Methods A case-control study was conducted among 125 neonates with CH (case group) and 375 neonates without CH(control group)in Fujian Neonatal Screening Center from January in 2012 to December in 2013. Univariate and multivariate logistic regression analysis were performed to identify the risk factors to CH during perinatal period. Results Univariate logistic regression analysis indicated that compared with control group,gestational hypertension,gestational diabetes mellitus,gestational thyroid disease and older age of mother were the risk factors to CH,the difference was statistically significant (P<0.05) and the risk of CH was higher in female babies,preterm babies,post-term babies low birth weight babies,macrosomia,twins,babies with birth defects and infection in cases group than those in control group,the difference was statistically significant (P<0.05). Multivariate logistic analysis showed that older age of mother(OR=2.518,95%CI:1.186-5.347), gestational diabetes mellitus(OR=1.904,95%CI:1.190-3.045),gestational hypothyroidism or hyperthyroidism(OR=12.883 and 30.797,95%CI:2.055-80.751 and 3.309-286.594),preterm birth (OR=4.238,95%CI:1.269-14.155),and post-term birth(OR=12.799,95%CI:1.257-130.327),low birth weight(OR=3.505,95%CI:1.059-11.601),macrosomia(OR=3.733,95%CI:1.415-9.851), twin or multiparous delivery(OR=5.493,95%CI:1.701-17.735),birth defects(OR=3.665,95%CI:1.604-8.371)and fetal distress(OR=3.130,95%CI:1.317-7.440)were the high risk factors to CH (P<0.05). Conclusion CH was correlated with mother’s age,gestational diabetes,gestational thyroid disease as well as neonate’s birth weight and gestational age,foetus number,fetal distress and other complicated birth defects at certain degree. More attention should be paid to perinatal care to reduce risk factors and the incidence of CH.

ObjectiveTo investigate the bactericidal effect of loaded multifunctional povidoneiodine-nanometer selenium （PVP-I@Se） disinfectant on Staphylococcus aureus （SA） and methicillin-resistant Staphylococcus aureus （MRSA）， and to provide an experimental basis for the reduction of surgical site infection （SSI）. MethodsThe control group was the povidone iodine （PVP-I） group with different concentrations of iodine （50， 75， 100， 200 and 400 μg/mL）. The PVP-I@Se group （experimental group） was the PVP-I group further supplemented with 2 μg/mL Selenium nanoparticles （SeNPs）. Then we compared the bactericidal effect of the two groups of disinfectant solutions on SA and MRSA by examining the minimum inhibitory concentration （MIC）， minimum bactericidal concentration （MBC）， the shortest sterilization time at a concentration of 50 μg/mL iodine and the inhibition zone diameters at concentrations of 200 μg/mL and 400 μg/mL iodine. ResultsMIC values of PVP-I against SA and MRSA were both 79.17 μg/mL， and those of PVP-I@Se were 54.17 and 70.83 μg/mL， respectively. MBC values of PVP-I against SA and MRSA were 129.17 and 150.00 μg/mL， respectively， and those of PVP-I@Se were 70.83 and 87.50 μg/mL， respectively. At a concentration of 50 μg/mL iodine， the shortest sterilization time of PVP-I for SA and MRSA was 130 s and 140 s， respectively， and that of PVP-I@Se was 65 s and 75 s， respectively. At a concentration of 200 μg/ml iodine， the inhibition zone diameters of PVP-I for SA and MRSA were 7.67 mm and 8.33 mm， and those of PVP-I@Se were both 9.50 mm. At a concentration of 400 μg/mL iodine， the inhibition zone diameters of PVP-I for SA and MRSA were 9.00 mm and 9.33 mm， and those of PVP-I@Se were 11.67 mm and 12.00 mm， respectively. ConclusionsPVP-I with different concentrations of 50， 75， 100， 200 and 400 μg/mL iodine supplemented with 2 μg/mL SeNPs have better and faster bactericidal effect on SA and MRSA. When combined with SeNPs， PVP-I can enhance the bactericidal activity against SA and MRSA， but with better sensitizing effect on SA than MRSA and higher demand of iodine concentration （400 μg/mL） for sensitizing effect on MRSA. This study provides a theoretical basis for selecting optimal concentration and action time of the disinfectant， thus reducing SSI.

OBJECTIVETo understand the relationship between perinatal factors and congenital hypothyroidism (CH) and provide scientific evidence for the prevention of CH.

METHODSA case-control study was conducted among 125 neonates with CH (case group) and 375 neonates without CH (control group) in Fujian Neonatal Screening Center from January in 2012 to December in 2013. Univariate and multivariate logistic regression analysis were performed to identify the risk factors to CH during perinatal period.

RESULTSUnivariate logistic regression analysis indicated that compared with control group, gestational hypertension, gestational diabetes mellitus, gestational thyroid disease and older age of mother were the risk factors to CH, the difference was statistically significant (P < 0.05) and the risk of CH was higher in female babies, preterm babies, post-term babies low birth weight babies, macrosomia, twins, babies with birth defects and infection in cases group than those in control group, the difference was statistically significant (P < 0.05). Multivariate logistic analysis showed that older age of mother (OR = 2.518, 95% CI: 1.186-5.347), gestational diabetes mellitus (OR = 1.904, 95% CI: 1.190-3.045), gestational hypothyroidism or hyperthyroidism (OR = 12.883 and 30.797, 95% CI: 2.055-80.751 and 3.309-286.594), preterm birth (OR = 4.238, 95% CI: 1.269-14.155), and post-term birth (OR = 12.799, 95% CI: 1.257-130.327), low birth weight (OR = 3.505, 95% CI: 1.059-11.601), macrosomia (OR = 3.733, 95% CI: 1.415-9.851), twin or multiparous delivery (OR = 5.493, 95% CI: 1.701-17.735), birth defects (OR = 3.665, 95% CI: 1.604-8.371) and fetal distress (OR = 3.130, 95% CI: 1.317-7.440) were the high risk factors to CH (P < 0.05).

CONCLUSIONCH was correlated with mother's age, gestational diabetes, gestational thyroid disease as well as neonate's birth weight and gestational age, foetus number, fetal distress and other complicated birth defects at certain degree. More attention should be paid to perinatal care to reduce risk factors and the incidence of CH.

Birth Weight ; Case-Control Studies ; Congenital Hypothyroidism ; epidemiology ; Diabetes, Gestational ; epidemiology ; Female ; Gestational Age ; Humans ; Hypertension, Pregnancy-Induced ; epidemiology ; Incidence ; Infant, Newborn ; Infant, Premature ; Maternal Age ; Neonatal Screening ; Pregnancy ; Pregnancy Complications ; epidemiology ; Premature Birth ; Risk Factors ; Twins