Objective To explore the locus of control and the relationship with parenting patterns of ado-lescents with first-onset major depressive disorder (MDD). Methods Patients with first-onset MDD (n= 98) and controls (n=98) were investigated by Sphere of Control Questionaire and EMBU, the results were compared and analyzed correlatively. Results The scores of the individual strength scale ( 37.47±13.66) and the relations with people scale ( 38.65±13.19 ) of patients were significantly lower(P<0.05 ) than the controls [(45.39± 12.84) and (45.90±11.20 ) each], while the scores of social-political management (39.01±10.28) were not significantly lower (P>0.05)than the controls' (39.12±9.50). The scores of patients' paternal excessive inter-ference factor(2.66±0.49) , maternal excessive protection/interference (2.79±0.55 ) and rejection/denial fac-tor(2.55±0.39) of EMBU were significantly higher(P<0.05)than controls' [(2.17±0.58),(2.17±0.49) and( 1.75±0.51 ) each]. The scores of patients' paternal affection/understanding factor were significantly posi-tively correlative with patients' scores of relations with people (r=0.27, P<0.05), maternal affection/under-standing factor was significantly positively correlative with patients' scores of individual strength (r=0.21, P <0.05) and relations with people(r=0.23, P<0.05 ) , while maternal excessive interference/excessive protection and individual strength (r=-0.24, P<0.05 ) and relations with people (r=-0.22, P<0.05), maternal rejec-tion/denial and individual strength (r=-0.26, P<0.05) were significantly negatively correlative. Conclusion Adolescents with MDD have abnormal locus of control,which might be correlative with their parenting patterns.

Objective To study the change of TLR4 on peripheral blood monoeytes (PBMCs) and its role in the pathogenesis of chronic severe hepatitis B. Methods The expression of TLR4 on 10000 CDI4 + PBMCs was determined by flow eytometer in 30 healthy control,31 patients with chronic hepatitis B and 30 patients with chronic severe hepatitis B. The level of serum tumor necrosis factor α(TNF- α) was determined by ELISA. Results The values of TLR4 on PBMCs and serum TNF-αof the groups of healthy control, patients with chronic hepatitis B and patients with chronic severe hepatitis B were 2.3±1.1,3.7±2.3, (6.9±4.1 ) mean fluorescence intensity (MFI) and (53.8±38.1 ), ( 164.3±89.9) and (359.8±140.0) ng/L. The TLR4 value in patients with chronic severe hepatitis B was signifi- cant higher than those in healthy control and the patients with chronic hepatitis B ( P <0.05). However, there was no significant difference between the patients with chronic hepatitis B and healthy control ( P > O. 05 ). TNF-α increased gradually and significantly from the healthy control to the patients with chronic hepatitis B and patients with chronic severe hepatitis B. There was a significant positive correlation be- tween the value of TLR4 and the value of serum TNF-αin the patients with chronic severe hepatitis B( r=0.666, P <0.01). Conclusion There may be a role of TLR4 in the pathogenesis of chronic severe hepatitis B.

Objective To study the effect of matrine on the expression of a proliferation-inducing ligand (APRIL) in colorectal cancer cell line (SW480 cell). Methods MTT assay was used to evaluate the inhibitory effect of matrine on SW480 cells. The protein and mRNA levels of APRIL in SW480 cells were determined by immunohistochemistry and real-time fluorescence quantitative PCR (RFQ-PCR). SW480 cells were treated with 0.5,1.0,2.0 mg/ml of matrine for 24 h, 48 h and 72 h. FU and blank were served as drug control and blank control groups, respectively. Results Matrine had obviously inhibitory effect on proliferation of SW480 cells in a time- and dose-dependant manner. The expression of APRIL was strong in SW480 cells. When treated with 50,100,200 ug/ml of FU, the APRIL mRNA levels in SW480 cells raised gradually and reached the highest levels at 72 h after treatment, which were significantly higher than those in blank control group (all P value<0.001). When treated with 0. 5,1.0, 2.0 mg/ml of matrine, the APRIL mRNA levels in SW480 cells increased at 24 h after treatment, which were significantly higher than those in blank control group (all P value<0. 001), and then decreased gradually and almost equal to level of blank control group at 72 h. Conclusion In treatment with FU, the survival cells.may have stronger ability of proliferation due to higher expression of APRIL in SW480 cells. Anti-APRIL therapy might be an important assistant treatment to counter the impact of APRIL. Matrine will not cause persistent increase of APRIL mRNA levels in SW480 cells, so it might be a helpful drug in anti-tumor theraphy.

AIM:To investigate the expression of TLR2 and TLR4 in hepatocellular carcinoma(HCC),and to analyze their correlations to clinicopathologic features of HCC.METHODS:The protein and mRNA levels of TLR2 and TLR4 in HCC and para-tumor tissue were determined by immunohistochemistry and real-time fluorescence quantitative PCR(RFQ-PCR).RESULTS:The protein and mRNA levels of TLR2 and TLR4 in HCC were lower than those in para-tumor tissue(P

Objective To observe the effects of 5-fluorouraeil(5-FU)and eisplatin(DDP)on the expression of Toll-like receptor 2 (TLR2)and Toll-like receptor4(TLR4)in hepatocellular carcinoma cell lines HepG2 and HepG2.2.15.Methods Direct immanotlaorescenee flow cytometry was used to detect mean flubrescence intensity(MFI)of TLR2 and TLR4,and TLR2 and TLR4 positive cell percentage in HepG2 and HepG2.2.15 cells before and after treated with 5-FU.and DDP at various concentrations for 24h,48h and 72h.Results MFI of TLR2 and TLR4.and TLR2 and 11LR4 positive cell percentage in HepG2.2.15 cells were significantly higher than those in HepG2(P＜0.01).After HepG2 and HepG2.2.15 cells were treated with different concentration of 5-FU and DDP,MFI of TLR2 and TLR4,TLR2 and TLR4 positive cell percentage in HepG2 and HepG2.2.15 cells almost had no change.only MFI of TLR2 in HepG2.2.15 cells decreased after cells were treated with 5-FU at the concentrations of 100,200μg/ml and DDP at the concentrations of 20μg/ml for 72h(P＜0.05 for all).Conclusions 5-FU and DDP can not activate TLR2 and TLR4 signal pathway in hepatocellular carcinoma cell lines HepG2 and HepG2.2.15.To find the activated pathway in TLR2 and TLR4 signal pathway,some other methods should be used,and this will be helpful in antieancer therapy.

Objective To study the clinical diagnostic value of transvaginal ultrasonic in early cervical carcinema and cervical neoplasis.Methotis The suspected patients with cervical carcinoma and cervical neoplasis were detected with transvaginal ultrasonography,liquid based cytology and cervical biopsy.The sonograms of transvaginal ultrasonic were retrospectively analyzed.Results In early cervical carcinoma and cervical neoplasis,the diagnostic sensitivity of transvaglnal ultrasonic was 90.9%and 83.3%;and the diagnostic specifity of transvaginal ultrasonic was 70.6% and 60.0%;the rate of missed diagnosis of transvaginal ultrasonic was 9.1% and 16.7%.Conclusion Transvaginal ultrasonic plays an important part in the clinical diagnosis of early cervical carcinoma.

Objective To investigate the role of TLR4 in the pathogenesis of chronic hepatitis B(CHB) by study the expression of TLR4 in liver tissues in patients with CHB, and the relationship among TLR4 and serum HBV DNA level, clinical severity degrees and histo-logical grades and stages. Methods Expression of TLR4 in liver tissues was semi-quantitatively determined by immunohistochemistry and e-valuated by a scoring system in 75 patients with CHB and 10 health controls. Results The positive staining of TLR4 mainly located in the cytoplasm and some on cell membrane of bepatocytes. Expression of TLR4 in the liver tissues of patients with CHB was stronger than that of health controls. The scores of TLR4 expression in patients with mild, moderate and severe CHB were 1.0±0.5,2.3±0.5 and 2.9±0.4. The scores increased gradually and significantly along with the increase of clinical severity degrees( F = 104.8, P<0.01). The scores of TLR4 expression in the liver tissues of patients with CHB were positively correlated with the clinical severity degrees (r=0.838, P<0.01) and histological grades (r=0.579, P<0.05), but not correlated with Lg (serum HBV DNA) or histological stages. Conclusion TLR4 was up-regulated in the hepatocytes of patients with CHB. There may be a role of TLR4 in the pathogenesis of CHB.

Objective To investigate the expression of CD40 and intercellular adhesion molecule-1 (ICAM-1) treated with lipopolysaccharide (LPS) in rat peritoneal mesothelial cells(RPMC) and the role of NF-κB signal transduction pathway. Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were exposed respectively to different concentrations of LPS for 12 h or treated with LPS (5 μg/ml) for different time points. To observe the effect of LPS on the expression of CD40 and ICAM-1, the RPMCs were treated with LPS (5 μg/ml) for different time points. To observe the effect of LPS on the expression of NF-κB and p-NF-κB protein, the RPMCs were treated by LPS or pretreated with BAY11-7085 (5 μmol/L or 1 μmol/L ) for 3 h, then treated with LPS for another 3 h, respectively. Expression of CD40 and ICAM-1 mRNA was examined by RT-PCR. Expression of NF-κB and p-NF-κB protein was detected by Western blot. Results Compared with medium control group, stimulation of RPMCs with 1 μg/ml and 5 μg/ml of LPS resulted in a significant increase in the expression of CD40 and ICAM-1 mRNA(P<0.05). 10 μg/ml of LPS had strongest effect on CD40 and ICAM-1 expression compared with that of 1 μg/ml and 5 μg/ml of LPS. Treatment with 5 μg/ml of LPS resulted in time-dependent increase in the gene level of CD40 and ICAM-1, with the peak at 3 h. However, after that time point, the gene level of them was gradually attenuated. Following treatment with LPS (5 μg/ml), the level of p-NF-κB began to increase at 15 min, gradually reached the peak at 1 h, and then decreased. But the level of p-NF-κB at 2 h was still significantly higher than that of medium control. 5 μmol/L of BAY11-7085 decreased significantly the up-regulation of CD40 and ICAM-1 induced by LPS. Conclusion LPS enhanced the expression of CD40 and ICAM-1 on RPMCs in a concentration-dependent and a time-dependent manner. LPS induced expression of CD40 and ICAM-1 depend on the NF-κB signal transduction pathway.

Objective Delayed xenograft rejection (DXR) is a major barrier to the long-term xenograft survial.This study evaluated the interaction between human peripheral blood mononuelear cells (PBMC) and porcine endothelial cells (PEC),and the effects of new generation of rabbit antihuman leukocyte polyclonal antibody (newRALG) inhibiting xenogeneic cell-mediated immune responses.Methods newRALG was obtained from rabbits after immunization with activated lymphocytes and monoeytes.PEC were isolated from aorta,and human PBMC were isolated from peripheral blood.Co-cultures of PKH-26 labeled PEC with PBMC were established,newRALG,thymoglobulin,isotype Ig and scavenger receptor (SR) ligand poly G were added into the co-cultures.Cells were collected,then FACS analysis was carried out to detect the up-take of PEC membrane by monocytes and the expression of costimulatory molecules.Lymphocyte proliferative responses to PEC with or without antibody were evaluated by a xenogeneie mixed lymphocyte-endothelial cell reaction (xMLER).Results FACS analysis revealed that monocytes from PBMC-PEC co-cultures became positive for PKH-26 following their interaction with PKH-26 labeled PEC,indicating that they engulfed PEC membranes during activation.PKH-26 positive monocytes up-regulated the CD40 and CD80 expression.Furthermore,SR blockade with poly-G prevented PEC membrane up-take by monocytes,newRALG greatly reduced SR-mediated PEC membrane up-take.The effects of thymoglobulin in inhibiting PEC membrane uptake were limited.xMLER demonstrated strong lymphocyte proliferation in response to PEC,and lymphocyte proliferation was dramatically inhibited by newRALG but not isotype Ig at a dosmdependent manner.Conclusions Monocytes play an important role in xenogeneic immune responses.SR ligand poly G inhibits PEC membrane up-take.newRALG inhibits PEC membrane up-take by monocytes,suggesting that newRALG blocks SR.Additionally,newRALG inhibits lymphocyte proliferation in response to PEC.These results suggest that this new polyclonal preparation may thus impair the initiation of xeno-specific immune responses and prevent xenograft rejection.

AIM: To investigate the dynamic variety of frequency and function of FoxP3~+ regulatory T cells in patients with acute hepatitis B (AHB). METHODS: Peripheral blood mononuclear cells (PBMCs) from 15 AHB patients at acute phase (week 1 of illness), convalescent phase (primary occurrence of both ALT level normalization and HBsAg negative conversion), resolved phase (at least 8 weeks after both ALT normalization and HBsAg seroconversion, and 15 health subjects were analyzed for FoxP3 (Forkhead/winged helix transcription factor) mRNA expression in MACS magnetic beads-purified CD4~+ T cells by real-time RT-PCR assay. The effects of Treg cells on the proliferation of CD4~+CD25~- T cells were examined by a ~3[H]-thymidine incorporation assay. RESULTS: AHB patients presented a significantly higher FoxP3 mRNA expression at convalescent phase than acute phase (t=-6.04, P<0.01) and resolved phase (t=4.45, P<0.01), and healthy controls (t=3.44, P<0.01). We also observed that the suppression efficiency of Treg cells on proliferation of CD4~+CD25~- T cells was lower at acute phase than convalescent phase (t=-5.30, P<0.01) and resolved phase (t=-3.20, P<0.05), but there was no significant difference between healthy controls and any phase of AHB. CONCLUSION: AHB patients presented lower circulating Treg frequency and suppression function at acute phase, and both of them are increased at convalescent phase, and then return to normal level along with disease resolved. This follow-up study furthers our understanding of Treg' s role in immunopathogenesis of hepatitis B.