Objective To develop a nested-real-time PCR method for the detection of FOXP3 mRNA in human peripheral blood mononuclear cells(PBMC).Methods The outer primers were designed by span intron,and the inner primers and TaqMan probe were designed according to the amplified sequence.During the nested real-time PCR,decrease primers concentration and cycle times were decreased in first PCR amplification to increase specific templates,then quantitative real-time fluorescence was detected in the second PCR amplification.The standard curve was created by FOXP3 and beta-actin recombined plasmid DNA respectively,and the results were presented as the ratios of FOXP3 mRNA to beta-actin mRNA.Results Total mean Ct(threshold cycle) in 6 tests was 14.66.The mean CV(coefficient of variation) was 5.81%.The standard curve showed a fine linear relationship between Ct and template concentration,and the correlation coefficient was 0.999.The sensitivity was 100 times higher than normal real-time PCR by reach 100 copy.Conclusions Nested-real-time-PCR for the detection of FOXP3 mRNA in human PBMC is a simple,repeatable,high specific and sensitive method to evaluate immune function and observe therapeutic effects in certain diseases for clinical routine application.

Objective To detect the ratio of CD4+ and CD8+ T cells and the number of CD4+CD25+ T regulatory cells(Treg)in human peripheral blood mononuclear cells(PBMC)of rheumatoid arthritis(RA)patients and evaluate the significance for diagnosis of RA.Methods The number of CD4+,CD8+ T cells and Treg cells were detected by flow cytometry and the ratio of CD4/CD8 was calculated.Results The ratio of CD4/CD8 and the number of CD4+CD25+ T cells detected in 49 RA patients were significantly higher than that in health group with statistical significance(P

An analytical method for determination of tetramethylpyrazine (TMP) in ligusticum chuanxiong hort by RP-HPLC coupled with countercurrent extraction used for the preconcentration of trace TMP was established. The column was packed with Zorbax ODS (250 mm×4 mm，5 μm). The mobile phase consisted of methanol water (1 % acetic acid)(35∶65)，the flow rate was 0.8 ml/min and detection was performed at 300 nm. The calibration curves showed good linearity over the range of 0.41 μg/ml to 4.1 μg/ml，γ＝0.9993，the recoveries were 92.3 %. The method was simple，fast，sensitive and accurate and could be applied to the analysis of ligusticum chuanxiong hort. The results showed that the content of TMP in ligusticum chuanxionghort were different with different sources.

Objective To observe the influence of the insulation blanket applied in general anesthesia on the incidence of postoperative delirium.Methods 60 patients with elective laparoscopic cholecystectomy under generalanesthesia with ASA of grade Ⅰ-Ⅱ were selected.According to the random number table,the patients were divided into group A(without insulation blanket)and group B (with insulation blanket),each group in 30cases.The dosage and drug batch number of anesthetic drugs were same.Evaluated each patient using the confusion assessment method-chinese revised (CAM-CR).Recorded the number of cases of delirium for each group when 24 hours,48 hours, 72 hours after operation.Results The incidence of postoperative delirium in group A was 6.7%,which in group B was 0.0%.The incidence of postoperative delirium of group B was lower than group A (P <0.05 ).Conclusion Using insulation blanket to protect body temperature in perioperative operation can reduce the incidence of postopera-tive delirium in perioperative operation.

Background:Helicobacter pylori( Hp)is the main cause of chronic gastritis( CG),and CagA and VacA are important pathogenic factors of Hp. Recent studies have indicated that CagA and VacA antibodies of Hp have close relationship with gastric cancer(GC)and peptic ulcer(PU). Aims:To investigate the relationship between different antibodies of Hp and GC,PU and CG in Xinjiang area. Methods:Eighty-four GC,108 PU and 196 CG patients from Jan. 2014 to Aug. 2014 were enrolled. 14 C-urea breath test was used to determine Hp infection. Hp was typed by immunoblotting technique. Relationship between different Hp antibodies and patients with GC,PU and CG was analyzed. Results:Type Ⅰ Hp-positive rate in GC,PU and CG groups was decreased stepwisely(47. 6% ,37. 0% ,21. 4% , respectively). The positive rates of Hp strains(CagA-128 kDa,VacA-110 kDa and UreB-66 kDa)differed significantly among GC,PU,and CG groups(P = 0. 013,P = 0. 011,P = 0. 002);GC was associated with CagA-128 kDa,VacA-110 kDa,VacA-90 kDa and UreB-66 kDa(P < 0. 05),PU was associated with VacA-110 kDa,UreB-66 kDa(P < 0. 05). UreB-66 kDa was the risk factor of PU and GC(P < 0. 05). Conclusions:GC is associated with CagA-128 kDa,VacA-110 kDa,VacA-90 kDa and UreB-66 kDa,PU is associated with VacA-110 kDa,UreB-66 kDa,and UreB-66 kDa is the risk factor of PU and GC.

Objective To explore the clinical experience of liver retransplantation in management of biliary complications after liver transplantation.Methods The clinical data of patients receiving liver transplantion in our department between April 2002 and Aug. 2004 were retrospectively analyzed, including 5 patients being subjected to liver retransplantation because of serious biliary complication.Results Three patients were cured with the survival time being 5 months, 6 months and 8 months respectively. Two patients died at 8th and 43rd day: one died of liver non-function combining with lung infection and heart failure, and another died of renal failure. Complications included liver absess, biliary infection, wound infection, hydrops abdominis and lung infection.Conclusions Liver retransplantation is an effective treatment for serious biliary complications after liver transplantation. Surgical procedure, indication, operative opportunity, perioperative monitoring and so on contribute to the increase of the survival rate of the patients receiving liver retransplantation.

Objective To isolate and purify the antioxidant compounds from fructus xanthii and to study their antioxidant activity.Methods Compounds were isolated by repeated silica gel,macroreticular resin Diaion HP-20 and Sephadex LH-20 column chromatography and their antioxidant property was determined by scavenging activity to DPPH and ?OH.Results Five antioxidant compounds were isolated from the ethyl acetate extracts of Xanthium sibiricum fruits and their structures were identified as 5,7,3',4'-tetramethoxyisoflavone,myricetin-3'-methyl ether,falcalindiol,1,8-heptadecadiene-4,6-diyne-3,10-diol and 3,4-dihydroxybenzoic acid.Among them,3,4-dihydroxybenzoic acid had the highest radical scavenging activity.Conclusion All above 5 compounds have high antioxidant activities,and antioxidant activity of 3,4-dihydroxybenzoic acid is the highest.

Objective To establish a method for extracting compounds in Fructus Xanthii by accelerated solvent extraction(ASE),and to simultaneously determine the compounds by GC-MS.Methods The extraction was performed by ASE in which the conditions were optimized by orthogonal test,then the compounds in different extracts with different polarity were determined by GC-MS.Results The optimized conditions of ASE for compounds in Xanthium sibiricum fruits were as follows: petroleum ether,ethyl acetate and methanol as extracting solvent in turn in 33 mL extracting tube and the volume of solvent being 65 mL;6 g sample,9 MPa of extracting pressure,90 ℃of extracting temperature,5 min of extracting time,two cycles and one time for each extraction.The GC-MS conditions were as follows: 1 ?L extract was analysed by GC-MS(Shimadzu Model QP-2010) instrument fitted with a flame ionisation detector,separation was achieved on a DB-5ms,DB-17ms and DB-WAX column(J&W Scientific,30 m,0.25 ?m id film thickness,1.4 ?m,USA) respectively,the column temperature was maintained at 60 ℃for 3 min and increased to 250 ℃,the scanning range was from m/z 40 to m/z 700(scanning interval: 0.5 s),and the compounds were identified by searching NIST and Wiley registry of mass spectral data.Conclusion There are 61 compounds found in the petroleum ether extract and in which 48 are identified,22 in the ethyl acetate extract and 18 identified,as well as 34 in the methanol extract and 29 identified with the methods of optimized ASE combined with GC-MS.

Aim To study the effect of Semen Sojae Preparatum isoflavone(SSPI)on proliferation and JAK2/STAT3 signal transduction pathway in rat vascular smooth muscle cell(VSMC)induced by Angiotensin Ⅱ(AngⅡ).Methods A cell proliferating model of VSMC induced by AngⅡ was established.The proliferation activity of VSMC was analyzed by MTT method.The expressions of angiotensin Ⅱ receptor 1(AT1R) were detected by RT-PCR method.The expressions of JAK2,STAT3 and phosphorylation protein were detected by Western blot.Results 100 ?g?L-1, 200 ?g?L-1 SSPI significantly inhibited the proliferation of VSMC induced by AngⅡ,and down-regulated the mRNA expression of AT1R.200 ?g?L-1 SSPI could significantly down-regulate the protein expressions of p-JAK2,p-STAT3.Conclusions The proliferation of VSMC induced by AngⅡcan be inhibited by SSPI.The mechanisms might be related to down-regulating the expressions of AT1R,and arresting the phosphorylation of JAK2/STAT3 signal transduction pathway.

OBJECTIVE To compare the treatment effects of secreory otitis media with artificial eustachian tube insertion and tympanotomy tube insertion.To provide useful information for the treatment of secretory otitis media.METHODS Seventy-one patients(86 ears) with secretory otitis media were randomly divided into tow groups:experimental group(36 cases,44 ears) and control group(35 cases 42 ears).The experimental group was treated by artificial eustachian tube insertion under nasal endoscope.Drugs were injected through the tube into the tympanic cavity everyday after flushing it. The control group were treated by traditional method—the tympanic ventilation tube.All patients were followed up for 6 months.RESULTS The experimental group: 21 ears(47.73%) were cured,19 ears(43.18%) were straightened up,and no effects observed in 4 ears (9.09%).The control group:12 ears(28.57%) cured,17 ears(40.48%) wered straightened up and no effects obsered in 13 ears(30.95%).The efficiency rate of the experimental group(90.91%) was much higher than the control group(69.05%,P