More and more research proved that CD4+CD25+ regulatory T cells (Treg cells) play an important role in immune tolerance in these years. Known as a T cell growth factor, Interleukin2 (IL-2) regulated the immune tolerance induced by Treg cells. IL-2 sustained the activity of Treg cells both in thymus and periphery while it was dispensable for the development of Treg cells in thymus. In addition, IL-2 signal affected the function of Treg cells and maintained their competitive fitness. Therefore, CD4+CD25+ Treg cells and IL-2 formed an immune network to regulate the processes of immune response, immune tolerance and autoimmune homeostasis.

OBJECTIVE:To establish a method for the contents determination of ginsenoside Rg1,ginsenoside Re,ginsenoside Rb1 and geniposide in Panax ginseng processed by Gardenia jasminoides. METHODS:HPLC was performed on the column of Agi-lent TC18 with mobile phase of acetonitrile-0.1% phosphoric acid,(gradient elution,ginsenoside Rg1,ginsenoside Re,ginsenoside Rb1)and acetonitrile-water(13∶87,V/V,geniposide)at a flow rate of 1.0 ml/min,the detection wavelength was 203 nm for ginsen-oside Rg1,ginsenoside Re,ginsenoside Rb1 and 238 nm for geniposide,column temperature was 40 ℃,and injection volume was 20 μl. RESULTS:The linear range was 0.418-3.762 μg for ginsenoside Rg1(r=0.999 8),0.270-2.430 μg for Re(r=0.999 8), 0.398-3.582 μg for ginsenoside Rb1(r=0.999 7)and 0.606-3.030 μg for geniposide(r=0.999 7);RSDs of precision,stability and reproducibility tests were less than 2.0%;recoveries were 97.86%-101.47%(RSD=1.29%,n=6),96.21%-100.11%(RSD=1.42%,n=6),96.24%-100.48%(RSD=1.57%,n=6)and 97.76%-102.44%(RSD=1.71%,n=6). CONCLUSIONS:The meth-od is simple with good precision,stability and reproducibility,and can be used for the contents determination of ginsenoside Rg1, ginsenoside Re,ginsenoside Rb1 and geniposide in P. ginseng processed by Gardenia jasminoides.

Glial cell line-derived neurotrophic factor(GDNF) is a kind of protein factor which can regulate the enteric nervous system in survival,differentiation,colonization and injury repair.It has been confirmed in the disorders of the enteric nervous system,such as hirschsprung and anorectal malformations,but the specific mechanism in regulation of this factor is still unknown.Studies have found that GDNF/GFRa1/RET and GDNF/GFRa1 / NCAM pathway may be involved in the growth and maturation of the enteric nervous system,the disorders of those pathways above may lead to diseases by affecting the differentiation,proliferation and migration of intestinal neural stem cells,causing dysfunctions in the anatomical structure and function of the intestinal neurons.

Objective: To evaluate the in vivo pharmacokinetics of strychnos alkaloids [brucine, total alkaloids in Strychni Semen (TASS), and optimal total alkaloids in Strychni Semen (OTASS)] by ig administration to rats. Methods: The rats were ig administered with strychnos alkaloids and their blood samples were taken. Huperzine A was used as an internal standard. The brucine in plasma of rats was detected by HPLC method. The compartment model was fitted and the pharmacokinetic parameters were calculated by DAS1.0 program. Results: The pharmacokinetic analysis showed that brucine behaved as a two-compartment model in the three solutions after ig administration in rats. Compared with brucine monomer administration, the solution of TASS and OTASS could obviously increase the absorption of brucine in vivo, but other pharmacokinetic parameters had no significant difference. Conclusion: After ig administration with strychnos alkaloids, the other alkaloids in Strychni Semen could promote the absorption of brucine.

Objective To establish the disease-syndrome integrated animal models suitable for the studies of TCM through differen- tiating the property of TCM syndromes of spontaneous diseased animal models.Methods With the observation on general behaviors, irritable degree,turning endurance time,pain threshold,urine and stools,luster of hair,growing speed of hair,body weight,tongue condition,degree of eyeball protruding,conjunctiva chroma,blood pressure,heart rate,etc.of spontaneous hypertension rats(SHR) and the comparison with normal rats,the study was carried out on the macroscopic description of property of TCM syndrome of SHR (14～18 weeks of age)and their ethology.Results The blood pressure of SHR at the early stage tended to raise with age growing. Compared with the normal group,the heart rate of SHR rats was obviously quicker(P

Objective To investigate the therapeutic effects of arsenic trioxide(ATO) plus vitamin K2(VK2) on proliferation of HL-60 cells from acute promyelocytic leukemia cell line and explore the possible mechanism.Methods ①HL-60 cells were exposed to ATO(0.0,0.5,1.0,2.0,4.0 μmol/L),VK2(0.0,2.5,5.0,10.0,20.0μmol/L),or both of different concentrations (0.5 μmol/L ATO + 2.5 μmol/L VK2,1.0 μmol/L ATO + 5.0μmol/L VK2,2.0 μmol/L ATO + 10.0 μmol/L VK2,4.0 μmol/L ATO + 20.0 μmol/L VK2) for 24,48 or 72 h,respectively.The method of CCK-8 was used to assess the proliferation of HL-60 cells and the half inhibitory concentration(IC50) of ATO or VK2 was calculated,respectively.②Combination index (CI) was used to evaluate the combinative effect of the two treatments:CI ＜ 1,=1 or ＞ 1 indicated synergistic,additive,or antagonistic effect,respectively.③After HL-60 cells were treated with 1.0 μmol/L ATO or 5.0 μmol/L VK2 individually or simultaneously for 48 h,Annnexin V/PI staining was performed to identify the apoptosis rate of each group.Untreated cells were used as control group.Results ①ATO or VK2 alone inhibited the proliferation of HL-60 cells in a concentration and time dependent manner.The IC50 of ATO or VK2 at time of 24,48,72 h were (22.86 ± 2.44),(6.66 ± 0.34),(4.14 ± 0.41) and (18.40 ± 1.12),(13.48 ± 0.73),(8.95 ± 0.40) μmol/L,respectively; ②The combination of ATO and VK2 illustrated a synergistic effect with CI ＜ 1.③No statistical difference was found among control group [(4.38 ± 0.56)％],1.0 μmol/L ATO group [(5.76 ± 1.63)％] and 5.0 μmol/L VK2 group [(6.38 ± 1.42)％] in the apoptosis rate(all P ＞ 0.05).However,the apoptosis rate of combined group did rise to (44.18 ± 8.42)％,with a significant improvement to that of VK2 or ATO group alone (all P ＜ 0.01).Conclusions The combination of VK2 and ATO exhibits an enhanced synergistical inhibitive effect on proliferation of HL-60 cells,and apoptosis may be involved in this synergy in part.

Aneurysm ; diagnosis ; Carotid Artery, Internal ; pathology ; Diagnostic Errors ; Female ; Humans ; Hypertrophy ; diagnosis ; Middle Aged ; Palatine Tonsil ; pathology

Objective To observe the expressions of DNA methyltransferases (DNMTs) 1,3a and 3b in retinoblastoma (RB).Methods Sixty-two RB samples and six normal retinas were studied,including 17 poorly differentiated and 45 well differentiated samples; 16 invasive and 46 non-invasive samples.The expressions of DNMT1,3a,and 3b,and Ki-67 were detected using immunohistochemical analysis.Brown staining of nuclei was considered to represent the positive stain for DNMT1,3a and 3b,and ki-67,blue staining as negative.The level of high expression of nuclear staining was,positive cells in DNMT1≥65 ％,in DNMT3a≥60％ and in DNMT3b≥40％.The correlations of DNMT1,3a and 3b expression in RB samples,and MIB-1 labeling index were analyzed.Results Viewed under the light microscope,negative expressions of DNMT1,3a and 3b were demonstrated in normal retinas,however,positive expression was observed in RB samples,with 100％ in DNMT1,98％ in DNMT3a and 92％ in DNMT3b.Comparing well differentiated RB samples with poorly differentiated samples,significant differences were found in high expression of DNMT1 (x2 =12.57,P＜0.05) and DNMT3a (x2 =10.54,P＜0.05) ; also in the positive cells of DNMT1 (U=179,P＜0.05) and DNMT3a (U=198,P＜0.05).No significant difference was found comparing high expression (x2=1.5,P＞0.05) and positive cells (U=307,P＞0.05) of DNMT3b.When comparing invasive tumor tissues with non-invasive tumors,significant differences were shown between high expression (x2 =4.72,P＜0.05) and positive cells comparing DNMT1 (U=236,P＜0.05).No significant difference was shown in high expression (x2=3.53,0.84; P＞0.05) in DNMT3a and DNMT3b,or in comparison with positive cells (U=338,257; P＞0.05).The expression of DNMTs was positively correlated with the MIB-1 labeling index in RB tissues (R2=0.554,0.376,0.219; P＜0.05).Conclusion There are high expressions of DNMT1,3a,and 3b in RB.

Objective To observe the expression and relationship of high-mobility group A(HMGA) 1,HMGA2,MIB-1 labeling index (LI) and let-7 in retinoblastoma (RB).Methods Forty-four RB samples were studied,including 11 poorly differentiated samples,33 well-differentiated samples; eight invasive and 36 non-invasive samples.The expression of HMGA1,HMGA2 and MIB-1 LI in RB were analyzed by immunohistochemitry.The HMGA1,HMGA2 were scored on a scale of 0 to high expression.0: no expression ; low: 1％ - 10 ％ ; medium: 11 ％ - 50 ％ ; high: ＞50 ％.The MIB LI were scored on a scale of 0to high expression.O: no expression;low: 1％ - 40％;high: ＞ 40％.Semiquantitative reverse transcription-polymerase chain reaction was used to assay the let-7 expression level: ≥ 80％ showed no significantly decreased expression; 60％ - 79％ showed medium decrease in expression; ＜ 60％ highly decreased in expression.Results In 44 RB samples,there were 14 cases with no HMGA1 expression (32％),11 cases with low expression (25％),10 cases with medium expression (23％),and nine cases with high expression (20％).Expression level of HMGA1 was significantly higher in poorly differentiated RB than in well-differentiated RB (x2 =11.3,P＜0.01) ; however,no statistically significant difference was found between invasive tumors and noninvasive tumors (x2 -5.9,P＞0.05).There were 11 cases with no HMGA2 expression (25％),11 cases with low expression-(25％),nine cases with medium expression (20％),and 13 cases with high expression (30％).Expression level of HMGA2 was significantly higher in poorly differentiated and invasive RB than in well differentiated and noninvasive RB respectively (x2=20.9,8.7; P＜0.05).There were 4 cases with no MIB-1 LI expression (9％),18 cases with low expression (41％),and 22 cases with high expression (50％).Expression level of MIB1 LI was significantly higher in poorly differentiated RB than in well-differentiated RB (t=5.2,P＜0.05).Higher expression of MIB-1 LI was found in invasive tumors than in noninvasive tumors,with no significant difference (t=-1.1,P＞0.05).Twenty seven cases had no significantly decreased expression of let-7 (61％).There were eight cases with medium decreased expression (18％) and nine cases with highly decreased expression (21％).Correlation analyses revealed that MIB1 LI expression significantly correlated with HMGA1and HMGA2 proteins (r=0.327,0.602; P＜0.05).A significantly inverse correlation existed between let-7 expression and HMGA1,HMGA2 proteins and MIB-1LI respectively (r=-0.247,-0.310,-0.392; P＜0.05).Conclusions Overexpression of HMGA1,HMGA2 and MIB-1 LI and down regulation of let-7 were demonstrated in RB.Supplying let-7 to RB cells can possibly inhibit HMGA1 and HMGA2 expression.