Objective To investigate the inhibitory effects of cationic polymeric liposomes (CPLs) vector-based RNA interference (RNAi) technology on the expression of rat MHCⅡ transactivator ( CⅡTA) and MHCⅡgenes .Methods According to the genetic information of CⅡTA downloaded from GenBank, three short hairpin RNA (shRNA) sequences targeting CⅡTA sequences were designed .CPLs vectors were constructed and coupled to shRNA plasmid vectors to form pCⅡTA-CPLs vectors .Six groups including control group , CPLs control group , pHK-CⅡTA control group and three pCⅡTA-CPLs groups were set up.Rat dendritic cells (DCs) were transfected in vitro.Real time PCR and flow cytometry analysis were used to detect the expression of CⅡTA and MHCⅡat mRNA and protein levels in DCs after transfec-tion.Results The pCⅡTA-CPLs vectors were successfully constructed .Compared with control groups ,the transcription level of CⅡTA and MHCⅡand the expression of MHCⅡat protein level were significantly in-hibited in all pCⅡ TA-CPLs groups ( P<0 .01 ) .The strongest inhibitory effects of pCⅡTA-CPLs on the ex-pression of CⅡTA and MHCⅡgenes were observed in the second pCⅡTA-CPLs group.There was a positive correlation between the expression of CⅡTA and MHCⅡ.Conclusion CPLs vectors were effective gene carriers.The constructed pCⅡTA-CPLs vectors significantly inhibited the in vitro expression of rat CⅡTA and MHCⅡ, which provided evidences for further investigation on pCPLs-CⅡTA vectors in vivo.

Objective To explore the clinical effect of the small incision with laser comprehensive treatment for patients with the great saphenous vein varices. Methods Small incision surgical ligation was done at the top and end of the great saphenous vein. Then took out the whole part or subsection of the great saphenous vein, with laser to treat the main part of the great saphenous vein and varicose vein group. Results 210 cases of 230 limbs all cured, cure rate was 100%. Conclusion The small incision with laser comprehensive treatment was an effective method for the great saphenous vein varices. It was worthy of clinical application and dissemination.

Objective To analyze postoperative complications and its prevention of small incision with laser comprehensive treatment the great saphenous vein varices. Methods Clinical data of 210 cases (total 230 limbs) in our hospital used small incision with laser comprehensive treatment the great saphenous vein varices were retrospectively analyze. Results 210 cases of 230 limbs,60 cases have concurrent subcutaneous congestion after surgery,6 cases have funicular induration,red and swallon,pain in the local crus,6 cases have skin paraesthesia due to saphenous nerve injure,4 cases have skin burns,2 cases have ulcer amalgamate infection. All of the cases have healed after the corresponding treatment. Conclusion This is a effective method for the great saphenous vein varices but may have complications. It should be taken seriously and processing.

Objective To establish the mouse bone marrow-derived dendritic cells expressing FasL protein and explore the mechanism of inducing allogeneic mouse spleen T lymphocyte apoptosis. MethodsMouse myeloid DCs were cultured in selective medium zontaining essential cytokines for DC growth in vitro. The mouse DCs were transfected with liposome-mediated FasL gene. The levels of FasL mRNA before and after transfection were assayed by real-time quantitative PCR. The expression levels of FasL protein were assayed by flow cytometry(FCM)and Western blot. Non-transfected DC，empty plasmid transfected DC and FasL transfected DC were infused intravenouslY into allogeneic mouse. After 7 days, the apoptosis in spleen T lymphocytes was evaluated by TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling)method and FCM. ResultsCultured in vitro, the mature myeloid DCs from mouse could be obtained.The expressions of FasL mRNA and protein in FasL transfected DCs were significantly higher. Through the detection of spleen T lymphocyte apoptosis with TUNEL，the apoptosis index(AI)was higher in FasL transfected DC(11.67±1.53)，compared with non-transfected DC(2.67±0.58)and empty plasmid transfected DC(3.33±0.58)，P＜0.01. ConclusionA large quantity of myeloid DCs can be obtained through in vitro culture in selective medium. The liposome-mediated FasL gene transfected DCs could successfully express high levels of FasL protein. Intravenous infusion of FasL gene transfected DCs could induce apoptosis of allogeneic mouse spleen T lymphocytes.

Objective To study the impact of narrow band ultraviolet B (NB-UVB) irradiation on the expression of T helper 17 (Thl7) and CD4,CD25 and regulatory T (Treg) cells and their related cytokines transfor-ming growth factor beta 1 (TGF-β1) and interleukin-6 (IL-6) to further clarify how NB-UVB treatment helps patients with psoriasis vulgaris.Methods Ninety patients with active stage and stationary stage psoriasis vulgaris (45 cases each) were treated with NB-UVB for 8 weeks.Fifty healthy persons were used as normal controls.Peripheral blood levels of Th17 and Treg were measured by flow cytometry.An enzyme-linked immunosorbent assay (ELISA) was used to measure the serum levels of TGF-β1 and IL-6 before and after treatment.Clinical efficacy was evaluated in terms of the area of psoriasis and severity index (PASI) scores.Restlts Before treatment,the patients showed significantly higher levels of Thl7 cells in their peripheral blood than the controls.Their ratios of Th17 to Treg cells and their serum levels of IL-6 were also significantly higher.The percentage of Treg cells and the serum level of TGF-β1 were significantly lower in the patients.After the NB-UVB treatment,the Th17 cells,the ratio of Th17 to Treg cells and IL-6 had all decreased significantly.The percentage of Treg cells and TGF-β1 levels were significantly high-er compared with before phototherapy.The total effectiveness rate was 86.7％,and the average PASI scores had de-creased significantly.The PASI scores were positively correlated with the percentage of Th17,the Th17 to Treg ratio,and the serum level of IL-6,and negatively correlated with the percentage of Treg cells and TGF-β1.Conclusion The imbalance between Th17 and Treg cells and their cytokines may play an important role in the pathogenesis of pso-riasis.NB-UVB is able to significantly down-regulate the levels of Th17,IL-6 and the progression of Treg levels and TGF-β1 expression.It can regulate the balance between Thl7and Treg cells,which may be one of the mechanisms of NB-UVB treatment for psoriasis.The clinical data demonstrate that NB-UVB treatment is a safe and effective therapy for psoriasis vulgaris.

Aim To investigate the method of cell culture for smooth muscle cells from rat cerebral basilar artery and understand cells growth and biological characteristics.Methods The explant attached method was applied for cell culture of rat basilar artery smooth muscle cells(BASMCs).The cultured BASMCs were identified by immunocytochemical staining.The activities of cells were indicated by the dynamic changes of intracellular calcium concentration observed by RF-5 000 fluorospectro-photometer.Results BASMCs grew out of tissue blocks by 5 days,reached confluency could be subcultured after 2 weeks.Cultured cells were identified by intensely positive immunocytochemical staining to smooth muscle actin-specific.Introduction of calcium channel agonists induced significant increase in Fura-2 fluorescence ratio(F340/F380)and cells were in good condiction.Conclusion Explant attached method is simple,efficient and economic.It provides an ideal cell model for the study of pathogenesis of the cerebral vascular diseases.

Objective To investigate the inhibitory effects of histidine grafted poly (β-amino es-ter) ( HGPAEs) vector-based RNA interference ( RNAi) on the expression of gene encoding myeloid differ-entiation factor 88 (MyD88) in rat liver tissues.Methods The sequence of small hairpin RNA (shRNA) was designed based on the genetic information of MyD 88.HGPAEs vector was constructed and coupled with shRNA plasmid targeting MyD88 to construct pMyD88-HGPAEs vector.Rats were divided into five groups including control group , HGPAEs treatment group , pHK-HGPAEs treatment group , shRNA treatment group and pMyD88-HGPAEs treatment group .The rats in each group were transfected with the corresponding inter-ventions through portal vein injection .Real-time PCR and Western blot assay were performed to detect the expression of MyD88 in liver tissues 3 days after transfection .Results The pMyD88-HGPAEs vector was successfully constructed .The expression of gene encoding MyD 88 was inhibited in rats from shRNA treat-ment group and pMyD88-HGPAEs treatment group (P<0.05).Significantly decreased expression of gene encoding MyD88 at mRNA and protein levels were observed in rats from pMyD 88-HGPAEs treatment group as compared with those from other groups (P<0.01).Conclusion HGPAEs vector might be used as a po-tential gene carrier .The expression of gene encoding MyD 88 in rat liver tissues could be significantly inhibi-ted through portal vein injection of pMyD 88-HGPAEs vector .This study provided evidences for further re-search on pMyD88-HGPAEs vector in a high responder model of rat orthotopic liver transplantation .

Objective To investigate the clinical value of serum Haptoglobin (Hp)levels in cervical cancer patients.Methods Serum Hp (with immunoturbidimetry method),SCC,CA125 (with electrochemiluminescence immunoassay method)level were detected in 45 patients with cervical cancer,37patients with cervical benign disease and 41 female healthy controls.Re-sults Serum Hp level of cervical cancer patients (mean:1.69 g/L)was strongly higher than the level of cervical benign dis-ease patients (mean:0.90g/L)and healthy subjects (mean:0.69 g/L)(t=3.26,4.42,P=0.001,0.000,respectively).Hp level rises highly in the early stage of cervical cancer.The area under receiver operating characteristic curve (AUCROC)of Hp was 0.853 which was higher than SCC (AUC=0.751)and CA125 (AUC=0.494).The sensitivity of Hp,SCC,and CA125 were 87%,49% and 29% respectively.The specificity of Hp,SCC and CA125 were 68%,88% and 83% respective-ly.Three tumor markers combined together could produce a positive detection rate of 93%.Conclusion The detection of Hp has clinic significance on diagnosis of cervical cancer,and it’s possible to become a serum tumor marker.

Objective To explore the CD38 expression in T-cens(CD38-T)of chronic lymphocytic leukemia(CLL).Methods Multi-parameter flow cytometry was used to detect the expression of CD38,ZAP-70 and CD4/CD8 in 83 CLL patients.Results CD38 was positively expressed in 49.4%(41/83)T-cells(CD38-T)and 50.6%(42/83)tumor cells(CD38-B)of all CLL patients.The expression of the CD38-T was highly correlated with CD38-B(r=0.553,P＜0.01).ZAP-70+CD38+-T and ZAP-70-CD38-T constituted 67.5%(56/83)of CLL patients.There was significant correlation between CD38-T and ZAP-70 (r=0.349,P＜0.01).There was 33.3%(14/42)patients with CD38-T in Binet A patients,and 65.9% (27/41)in Binet B+C patients.There was also significant correlation between CD38-T and Binet B+C(r=0.312,P＜0.05),and CD4/CD8(r=0.453,P＜0.05).Conclusions Aberrant expression of CD38 in T-cells might be of prognostic relevance in CLL Most of the patients with high level CD38-T expression are accompanied with disorder or imbalance of immune function.CD38 expression in T cells might be a new prognostic factor in CLL.