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.

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 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.

ObjectiveTo evaluate the feasibility and efficacy of ex-vivo liver resection combined liver autotransplantation for patients with massive primary liver cancer who underwent complex liver resection.Methods The clinical data of 4 patients suffering from massive primary liver cancer who were admitted to the Beijing Chaoyang Hospital from January 2008 to May 2010 were retrospectively analyzed.Regular liver resection could not be carried out because the first,second and third hepatic hilum of the 4 patients were invaded by the tumors,so ex-vivo liver resection combined liver autotransplantation were performed.ResultsThe operation was successfully carried out for the 4 patients.The operation time,the duration of anhepatic phase and the volume of operative blood loss were 690-840 minutes,250-300 minutes and 400-1400 ml,respectively.Portacaval bypass operation was not performed.After ex-vivo liver resection,the inferior vena eava or hepatic vein and portal vein of the 4patients were repaired,and the allogenous blood vessels were kept to extend the superior vena cava of the remnant liver so as to facilitate the anastomosis of blood vessels and reconstruction of the first hepatic hilum. After operation,the hepatic function of 1 patient was back to normal; 1 patient who stfffered from abdominal hemorrhage received reoperation for hemostasia; 1 patient was found with hepatic dysfunction; 1 patient died of hepatorenal dysfunction at postoperative day 5.Compensatory hypertrophy was observed in the 3 patients who survived at postoperative months 1-2.Of the 3 patients,2 were found with multiple pulmonary metastases at postoperative months 8 and 9,and they died at postoperative mouths 13 and 15.Until April 2012,1 patient survived for 37 months with no tumor recurrence or metastasis. ConclusionsEx-vivo liver resection combined liver autotransplantation provides the technical feasibility for performing complex liver resection for patients. The incomplete compensation of liver function and the short-term recurrence of tumors after operation are still the main issues which hinder the development of this technique.

Objective To investigate the curative effect of percutaneous transhepatic cholangial drainage(PTCD) on patients with non-anastomotic stricture after liver transplantation when treated with endoscopic retrograde cholangio-pancreatography(ERCP) unsuccessfully.Method The clinical data of 29 patients with non-anastomotic biliary stricture after liver transplantation were retrospectively analyzed,who failed to respond to ERCP and underwent PTCD from January 2005 to December 2007.Result All patients were performed PTCD successfully including cholangiography in 141 cases,drainage tube replacement in 115 cases,and balloon dilation of bile duct stricture in 39cases.The intubation time ranged from 2 months to 65 months.The mean levels (x ± SD) of alanine aminotransferase,aspartate aminotransferase and total bilirubin were 68.0 ± 29.1 U/L,52.6 ± 34.8 U/L,63.2 ± 33.3 μmol/L after treatment in comparison to 178.3 ± 63.3 U/L,144.0 ± 59.1 U/L,154.2 ± 92.0 μmol/L before treatment.Conclusion PTCD,which could improve the symptoms and prolong the survival time of both grafts and patients in spite of inconvenience of intubation,is suggested for patients with non-anastomotic biliary stricture if they are not suitable for liver retransplantation.

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 investigate the expression of peripheral blood (PB) CD4+ CD25+ Foxp3+ regulatory T cells (Tregs) in patients with benign end-stage liver disease after liver transplantation and the relationship between levels of PB Tregs and acute rejection. Methods A prospective analysis was performed on 55 consecutive patients who underwent liver transplantation.Fourteen out of 55 cases suffered from acute rejection after liver transplantation were defined as rejection group,while the rest patients were classified into no acute rejection group. PB was obtained from liver transplant patients at different time points longitudinally: pre-transplant, post-transplant within one year and acute rejection. The circulating CD4+ CD25+ Foxp3+ Tregs in PB were measured by flow cytometry. Blood samples were drawn during acute rejection, at the same time, liver biopsies were performed. The circulating CD4+ CD25+ Foxp3+ Tregs were compared between two groups.Results There was no difference between two groups in levels of circulating CD4+ CD25+ Foxp3 + Tregs cells pre-transplant. However, the levels of circulating CD4+ CD25+ Foxp3+ Tregs in rejection group were decreased significantly as compared with no-rejection group (2. 23 % ± 0. 54 % vs. 2. 99 % ±0. 86 %,P＜0.01). The frequency of CD4+ CD25+ Foxp3+ T cells was negatively correlated with rejection activity index (RAI) (r = - 0. 80, P＜0. 01 ). Conclusion Monitoring PB CD4+ CD25+ Foxp3+ Tregs levels may be helpful in evaluating the immune state and act as a more sensitive marker for acute rejection diagnosis in the patients following liver transplantation.

Objective To study the mechanism of unscheduled Cyclin B1 expression at G_1 phase which is usually at G_2/M phase.Methods Human peripheral blood lymphocytes(PBL) from healthy volunteers were firstly activated by PHA and then went into cell cycle.The cells were collected at 0,36,48 and 60 h after activation and divided into two parts:one for Cyclins/DNA muhiparameter assay,and another for Post-sorting Western blot.Results After activation by PHA,Cyclin B1 and CDK1 of lymphocytes were expressed at G_1 phase.Conclusion Unscheduled Cyclin B1/CDKl probably contributes to lymphocytes in vivo into cell cycle.