Acute rejection is one of the serious early postoperative complications after liver transplantation.Many studies have shown that acute rejection was mainly mediated by T cells,while humoral factors were responsible for chronic rejection.However,accumulating evidences have demonstrated that humoral immune factors also played an important role in early acute rejection and usually resulted in severe adverse events.Here we clarify the role of humoral immunity in liver transplant rejection,which may help guide the clinical management of such patients with humoral rejection after liver transplantation.

OBJECTIVE:To isolate the chemical constituents from Radix isatidis.METHODS:The 95% ethanol extract of Radix Isatidis underwent adsorption by silica gel,and the portion eluted by different solvents was isolated and purified on silica gel column repeatedly.The physico-chemical constants and spectral data of the obtained compounds were determined and their chemical structure was identified.RESULTS:6 compounds were separated from chloroform and n-Butanol parts of Radix Isatidis and identified as:cholesterol(Ⅰ),indigotin(Ⅱ),indirubin(Ⅲ),p-Hydroxybenzaldehyde(Ⅳ),salicylic acid(Ⅴ),lariciresinol-4,4'-2-O-?-D-glucopyranoside(Ⅵ),respectively.CONCLUSION:For the first time,compound(Ⅰ)and compound(Ⅳ)were isolated from Radix Isatidis,this finding serves as reference.

OBJECTIVE:To extract and separate the water-soluble chemical constitutions from Radix Isatidis.METH ODS:The Radix Isatidis water extract underwent adsorption by D 101 macroporous resin,the portion eluted by ethanol of different concentrations was isolated and purified on silica gel column repeatedly,the physicochemical constants and the spectra data of the compound obtained were determined and its chemical constitutions were identified.RESULTS:3compounds temporarily separated from Radix Isatidis water extract,were identified as:syringin(Ⅰ),indole—3—acetonitrile—6—O—?—D—glucopyranoside(Ⅱ)and(+)—isolariciresinol(Ⅲ),respectively.CONCLUSION:For the first time,compound(Ⅰ)and compound(Ⅱ)were isolated from Radix Isatidis.

Objective To explore the early clinical characteristics and influencing factors in children with autism spectrum disorders(ASD).Methods From January 2005 to December 2014,193 children with ASD were collec-ted by continuous grouping method from Children's Rehabilitation Training Center in Xi'an.According to the 1∶1 matched case-control study requirements,and the other 193 children from kindergartens and primary schools in the urban areas of Xi'an were collected as healthy control group from March 1 to July 1,2016.The age of children in the case group was(40.78±14.86)months and the age of the healthy control group was(40.61±14.40)months.There were 167 boys and 26 girls in 2 groups and the ratio of boys to girls was 6.42∶1.00.The general status questionnaires,medical history questionnaire,diagnostic chart,Autism Behavior Checklist(ABC)and Family Environment Scale of Chinese version(FES-CV)were completed by parents between 2 groups.Childhood Autism Rating Scale(CARS)was completed by doctors in the case group.By using Excel software,the original questionnaires were completed in 2 entries by 2 persons to set up the database.All data were analyzed by SPSS 17.0 statistical software and conditional Logistic regression was used for multivariate analysis.Results Seventy point eight percent(137/193 cases)of children with ASD had been found abnormal under 2 years old or at 2 years old,and 54.9％(106/193 cases)had been diagnosed under 3 years old or at 3 years old.The average delay from the discovery to the diagnosis was 17 months.The initial abnormalities appea-rances were mainly manifested as no response to calling in 153 cases(79.3％),very little active contact with others in 141 cases(73.1％),silent or less use of oral language in 137 cases(71％),avoiding contact with the eyes of others or lack of facial expressions in 121 cases(62.7％).Their signs were easy to be misdiagnosed as mental retardation and language retardation.Children in the case group began to walk alone at the age of 8 months to 3 years old,and only 62.2％(120/193 cases)of them could walk alone at the age of 18 months or before.The age of conscious speech was at 8 months to 4 years and 4 months,and only 39.4％(76/193 cases)of the ASD children could speak at the age of 18 months or before.The total scores of the ABC scale of the case group were(56.520±22.140)scores and the sub-scales and total scores were significantly higher than those of the healthy control group,and the difference was statistically significant(t=16.845,27.390,16.527,26.320,23.371,32.206,all P<0.001).The positive consistent rate of ABC and clinical diagnosis was 56.5％.The total scores of CARS in the case group was(36.4±8.6)scores,and the positive consistent rate of CARS and clinical diagnosis was 78.8％.There was a statistical significance between the 2 groups in parental education,mother's occupation,family history(x2=29.670,44.593,15.439,6.095,all P<0.05),and there were statistical significance in the main caregivers,family harmony and family income(x2=19.006,7.129,109.027,all P<0.05).There was no statistical significance between the 2 dimensions of independence and achievement orientation between the 2 groups(t=-1.559,-0.139,P=0.120,0.890).The case group in the family cohesion,expressiveness,intellectual-cultural orientation,active-recreational orientation,moral-religious emphasis,organization and control of the 7 dimension scores were significantly lower than those in the healthy control group,and the differences were statistically significant(t=-7.683,-5.734,-8.762,-14.109,-2.026,-4.530,-2.464,all P<0.05).In the case group,the scores of the conflict dimension were higher than those of the healthy control group,and the difference was statistically significant(t=4.925,P<0.001).There was a statistical significance between the 2 groups in gestational age and birth hypoxia(x2=6.898,27.180,all P<0.05).According to multivariate analysis of Logistic regression,people other than parents serving as the primary support,anoxia of newborn,mother of non professional and technical personnel and lower scores of family active-recreational orientation might be the risk factors of ASD,family per capita income of 3 000 Yuan RMB or more monthly,mother education level of high school and above,and lower scores of family conflict might be the protective factors for ASD.Conclusions Clinical features of most ASD children can be easily identified under 2 years old,but if the diagnosis is delayed,the related intervention is late,so importance should be attached to early diagnosis.Mother's occupation and education level,family economic status,family environment,their supervisors,and anoxia of newborn may be the effective entry points in the prevention and treatment of ASD.

Objective To explore the relationship among the expression level of MMP?13, AGG and Col?II in the chon?drocytes caused by nutritional deficiencies in rabbits. Methods 30 New Zealand white rabbits were randomly divided into autolo?gous chondrocyte transplantation group (control group, n=10), nutrition block group (surgery group, n=10), and peripheral nutrition block group (sham surgery group, n=10). 4 weeks after treatment, the rabbits were sacrificed for undergoing the observations on general and histological level;real?time PCR assay was employed for testing the expression level of MMP?13, AGG and Col?II;cel?lular apoptosis percentage was observed through TUNEL stain. The relationship among the apoptosis level, cartilage cells histologi?cal Mankin score as well as the expression level of MMP?13, AGG and Col?II were analyzed. Results Based on the Mankin score, there was a statistic difference between surgery group and control group. On the other side, there were no statistic differenc?es between sham surgery group and control group. 4 weeks after treatment, surgery group presented a higher apoptotic percentage compared with control group;this value between sham surgery group and control showed no significant differences. There was an increased mRNA expression level of MMP?13 and a decreased mRNA expression level of AGG and Col?II in surgery group com?pared with control group;no statistic differences of all these values was found between sham surgery group and control group. His?tological Mankin score and apoptotic percentage presented positive correlation (r=0.922, P<0.001), the regression equation:Y=-0.548+0.404X, R2=0.844 (F=157.735, P<0.001); the mRNA expression level of MMP?13 and apoptotic percentage presented positive correlation (r=0.942, P<0.001), the regression equation:Y=0.951+0.116X, R2=0.883 (F=219.054, P<0.001). There was a nega?tive correlation between the mRNA expression level of MMP?13 and the mRNA expression level of AGG as well as Col?II (r=-0.956,-0.945, P<0.001). Conclusion Damage of cartilage cells causes the up?regulation of the MMP?13 expression which could ex?acerbate the degeneration of cells. It could induce the down?regulation of AGG and Col?II mRNA expression, which will cause the extracellular matrix synthesis disorder.

Objective To assess the impact of miR-429 on lung adenocarcinoma cell SPC-A1 growth inhibition.Methods Pre-miRTM miR-429 precursor was synthesized and transfected to the SPC-A1 cells by liposome; qRT-PCR assay was used to quantify the miR-429 expression levels; The proliferation of SPC-A1 cells was evaluated by Cell Counting Kit-8 (CCK8).The cell apoptosis was evaluated by Annexin V Assay; The cell cycles of each group were assayed by flow cytometry;Western-blot was used to analyze the expression of cylines.Results The expression level of miR-429 was highly induced after transfection (P ＜ 0.001) ;CCK-8 assay showed the cell proliferation activity of pre-miR-429 group was lower than that of blank and control group 48h and 72 h after transfection(P =0.0167,0.0383,P =0.0320,0.0465),whereas the apoptosis rate had no significant difference between pre-miR-429 and control 24h after transfection by Annexin V Assay(P ＞ 0.05) ; The flow cytometry at 48h after transfection showed that miR-429 decreased the percentage of cells in G1 phase,but increased in S phase,indicating the cell cycle arrest at S phase(P =0.0010,0.0010 ; P =0.0068,0.0133) ; however,the expression level of Cyclin E in pre-miR-429 group had no difference compared with control.Conclusion miR-429 could inhibit cell proliferation and promote cell cycle arrest of lung adenocarcinoma cell SPC-A1.miR-429 may play a potential tumor suppressor role in lung adenocarcinoma cell SPC-A1.

Objective To study the clinical significance of CD39 on regulatory T (Treg) cells in the peripheral blood (PB) of patients following liver transplantation and to evaluate the relationship between the levels of CD39+ Treg cells in the PB and acute rejection.Methods A prospective study was conducted to compare the CD39+ Treg cells from 76 liver transplant patients with those coming from 20 age-matched healthy individuals.The PB samples were collected within one year at different time points post-transplant.Blood samples and liver biopsies were collected at the time when acute rejection was diagnosed.The percentages of CD39 within the CD4+ CD25+ T cells were measured by using flow cytometry.The liver transplant patients were classified into two groups:the rejection group which consisted of 17 patients who an episode of acute rejection,and the non-rejection group consisted of the remaining 59 patients who had no acute rejection episodes.The percentages of CD39 within the CD4 + CD25 + T cells and the inhibition function of the CD39+ Treg cells were compared between the two liver transplant groups.Results The percentages of CD39 within the CD4+ CD25+ cells were significantly lower in the rejection group during acute rejection as compared to the non-rejection group (P ＜ 0.05).The percentages of CD39 within the CD4 + CD25 + cells were negatively correlated with the Rejection Activity Index (r =-0.86,P ＜ 0.05).The inhibition rate regarding the CD4+ CD25+ CD39+ Treg cells in patients with acute rejection was significantly lower than those without rejection (P ＜ 0.05).Conclusions The percentages of CD39 within the CD4+ CD25+ T cells were significantly lower in the rejection group during acute rejection and were negatively correlated with the RAI.The inhibition rate regarding the CD4+ CD25+ CD39+ Treg cells in patients with acute rejection was significantly lower than those without rejection.

Objective To investigate the partial safety of the recombinant adenovirus containing the triple-point mutants HIF-1αgene (Ad-HIF-1α-Trip)transfection in a rabbit model of hind limb ischemia. Methods After ligation of left femoral artery, 22 New Zealand whites rabbits were randomly divided into 3 groups: saline group(n=6), Ad-Null group(n=6) and Ad-HIF-1α-Trip group(n=10). After operation, saline, Ad-Null and Ad-HIF-1αwere injected intramuscularly respectively. The expression of transferred HIF-1αat mRNA level in the ischemic skeletal muscle and other important organs were detected by Real-time PCR 10 days after gene transfection. The body temperature, weight, blood, liver and renal function, as well as the myocardial enzymes were detected before operation, and on the 3th, 7th, 14th and 28th day after gene transfection, so that pathological changes could be observed. Results On the 10th day after gene transfection, obvious expressions of HIF-1αat mRNA level in the ischemic limb were found, but no expression in other important organs was detected in Ad-HIF-1α-Trip group. The blood routine, liver and renal function were all in the normal range (P > 0.05). No abnormalities were found in heart, liver, kidney, and lung HE transfection in Ad-HIF-1α-Trip group. Conclusion Single intramuscular injection of Ad-HIF-1α-Trip can be expressed obviously in the ischemic limb without detected damage of liver , cardiac and kidney.

A method for simultaneous determination of 20 Perfluorinated alkyl substances ( PFAS) in animal liver using QuEChERs and HPLC_MS/MS technique was developed. The samples were extracted with 0. 1%hydrochloric acetonitrile and cleaned up with C18 , N_Propylethylendiamine ( PSA ) and graphitized carbon blacks ( GCB ) . The analytes were separated by a reversed phase C18 column and gradiently eluted with a mixed solution of 5 mmol/L ammonium acetate methanol and 5 mmol/L ammonium acetate. The samples were quantified using isotope internal standard and external standard with the matrix matched standard calibration curve method. Good linearity was obtained for all the 20 PFAS at the concentration of 0. 1-10 μg/L with the linear correlation coefficients more than 0. 9995. The limits of detection (LOD) and the limits of quantification ( LOQ) for PFAS were 0. 05-0. 2 μg/kg and 0. 4-0. 5 μg/kg, respectively. The recoveries at three different concentration levels ( 0 . 5 , 2 and 5 μg/kg ) were in the range of 70 . 3% -108 . 1%. The repeatability expressed as relative standard deviations (RSD) was ranged from 2. 1% to 11. 9% (n=6).

OBJECTIVE:To study the activity of 10 chemical parts extracted from Isatis indigotica against herpes simplex virus(HSV)in vitro and to determine the antiviral active parts.METHODS:10 crude fractions and refined portions were extracted from Isatis indigotica using solvent isolation and absorption chromatography.Their inhibitory effects against HSV-Ⅰ and HSV-Ⅱ were determined by MTT using virus inhibition ratio and therapeutic index(TI)as appraisal targets to compare the antiviral effects among all the chemical parts.RESULTS:The parts(Ⅷ,Ⅹ)with EtOH(10% and 50% in concentration)elution via macroporous resin absorption showed the most potent activities,whose inhibitory rates against HSV-Ⅰ were 53.21% and 56.28% respectively,similar the action of its positive control acyclovir(62.55%),and their TI were 4.61 and 18.62 respectively.CONCLUSION:There were significant differences in the antiviral activity among all the chemical parts extracted from Isatis indigotica,