Objective To investigate the effect of rosiglitazone on the CD4+regulatory T cells in the patients with latent autoimmune diabetes in adults(LADA).Methods The CIM+T cells from IADA patients were isolated with anti-CD4-dynal magnetic beads.The expression of Foxp3 mRNA,along with peroxisome proliferators activator receptors gamma(PPARγ)mRNA and TGF-131 mRNA was determined.The effect of rosiglitazone on CD4+T cells was measured,after treated with rosiglitazone for 48 h.Cell viability was assessed by Mtit assay.The proliferation was assayed with 3 H-TdR.Two-color staining(anti-CD4,anti-CD25)flow cytometric analysis was employed to measure the percentage of CD4+CD25+T cells of Deriph eral blood.Resuits PPARγmRNA was expressed in peripheral CD4+T lymphocytes.RosiglitazoBe inhibited phytohemagglutinin(PHA)-induced human CD4+T cell proliferation in dose dependence.The percentage of CD4+CD25+T cells showed no significant change after the peripheral blood culture with 1 μmol/Land 10μmot/L rosiglitazone.10 μmol/L of rosiglitazone induced Foxp3 mRNA expression in vitro (3.27fold,P<0.05),whereas TGF-β1 mRNA expression did not change.Furthermore,only 1 μmol/L of rosiglitazone could promote Foxp3 mRNA expression if adding IL-2(10 U/m1)in cultures(3.48 fold.P

Objective To develop a new system of computer-aided design(CAD) for individual metacarpophalangeal joint prosthesis based on rapid prototyping technique. Methods A hand of cadaver was scanned with PLUS-4 spiral computed tomography (CT).Then the transaxial 2D image data of digitus metacarpophalangeal joint were reconstructed into 3D digitized contour data by 3DMSR that was designed by ourselves. Then,the intramedullary stem was designed in software of Surface 9.0. Results The 3D contour image of metacarpophalangeal joint presented was reconstructed by 3DMSR and edited by Surface 9.0 easily for CAD of individual metacarpophalangeal joint.The intramedullary cavity was like choanoid. The intramedullary stem longitude of articular head and fossa were 47.31 and 35.20 mm.The intramedullary stem fit cavity.The model fit anatomical shape. Conclusion The 3D contour image of metacarpophalangeal joint can be obtained by spiral CT scanning,and the digitized data can be applied directly to CAD of individual artificial joint and subsequently rapid prototyping fabricating.In addition,the reconstruction method is simple and can be applied widely to clinical implant fabricating practice of orthopaedics.

Objective To discuss the anticancer role of arsenic trioxide (ATO) with cisplatin on human oral carcinoma HSQ-89 cells. Methods The human oral epidermoid HSQ-89 cells were chosen as the subjects. Different concentrations of ATO were added into Cisplatin(DDP)-treated cells. The inhibition rate of tumor cells was detected by MTT assay. Results Different concentrations of ATO (0,2.5,5,7.5,10,12.5 μmol/mL) were added into oral cancer HSQ-89 cells which have been treated with DDP (15 μg/mL). The inhibition rate of tumor cells were 26.9%, 67.5%, 73.0%, 88.5%, 90.4%, 98.7%respectively; The combined application of ATO with cisplatin could improve the inhibition rate of HSQ-89 cells in a dose-dependent relation. Conclusion The combined application of ATO and DDP can produce a synergistic action of inhibition on oral cancer cell.

Objective Paraquat(PQ) is an effective herbicide which is widely used in agricultural production .PQ poisoning is frequently seen in humans with the lung as the target organ ,but the poisoning mechanisms is not very clear .Studies show that endoplasmic reticulum stress(ERS) is closely associated with poisoning , but there are few reports on the relationship between ER stress and PQ poi-soning.This article was to investigate the effects of ERS-induced apoptosis and total flavonoids from astragalus complanatus (FAC) in a-cute lung injury(ALI) following paraquat poisoning in rats . Methods A total of 30 adult healthy Sprague-Dawley (SD) rats were ran-domly divided into 3 groups:control group, ALI group, ALI+FAC group and ALI+saline group.Biochemical method was applied to de-tect superoxide dismutase (SOD), catalase (CAT) and malondialdehyde (MDA) in lung tisssue,TdT-mediated dUTP nick end labeling (TUNEL) assay in observing lung apoptosis, Western blotting and real-time PCR(RT-PCR) in detecting the changes in expressions of C/EBP homologous peotein (CHOP), activating transcription factor 4 (ATF4) and X-box binding protein 1 (XBP) following ALI, and HE staining in observing the pathological changes of lung tissue . Results Compared with control group , the expression of MDA content was enhanced in ALI group ([3.26 ±0.24] vs [5.04 ±0.36],P<0.01), along with significantly decreased activity of SOD and CAT ([300.26 ±35.69] vs [187.21 ±25.66]), ([5.78 ±1.28] vs [2.15 ±1.12],P<0.01), increased cell apoptosis , upregulated pro-tain level of CHOP ([0.74 ±0.20] vs [0.23 ±0.07],P<0.01) and mRNA expression of ATF4, XBP1 and CHOP.However, FAC sig-nificantly attenuated ALI following PQ , as showed by reduced MDA content , enhanced activity of SOD and CAT , decreased cell apopto-sis, inhibited protain level of CHOP and mRNA expression of ATF 4, XBP1 and CHOP ([5.04 ±0.36] vs [3.99 ±0.27],P<0.01). Furthermore, the activity of SOD and CAT were higher in FAC pretreatment group than those in ALI group ([ 0.74 ±0.20 ] vs [0.42 ±0.11],P<0.01). Conclusion From the research, ERS-induced cell apoptosis is involved in ALI following PQ , and the protec-tive role of FAC in lung tissue following PQ is due to its effect in atten-uating ERS-induced apoptosis .

OBJECTIVE: To establish a method for the determination of the peroxide value of hydroxycamptothecin freeze-dried liposome. METHODS: Colorimetric method with ferric thiocyanate and UV spectrophotometry were adopted for the content determination of peroxide value at detection wavelength of 500 nm. RESULTS: The linear range of iron was 0.2～4 ?g?mL-1 (r=0.999 0) with an average recovery of 99.2% (RSD=1.2%, n=9). The peroxide value of 3 batches of hydroxycamptothecin freeze-dried liposome was 0.48,0.45,0.43 mmol?kg-1. CONCLUSION: The method is sensitive and precise for the determination of the peroxide value of liposome.

AlM: To study the effects of different concentrations of Coix seed oil on human retinal capillary endothelial cells ( HRCECs ) proliferation and vascular endothelial growth factor ( VEGF) expression in high glucose environment.
METHODS: HRCECs extracted from human fresher eyeball and cultured in vitro, and ultimately used in the experiment were the growth of 3rd ~ 4th cells, the experimental were divided into blank control group, low glucose control group, high glucose control group, high glucose + ( 50ü L/mL, 100ü L/mL, 200ü L/mL ) different concentrations Coix seed oil group. Detecting the multiplication of HRCECs by MTT, the immunocytochemical method was employed to detect the each group HRCECs of VEGF expression.
RESULTS:MTT assay results showed that: different concentrations of coix seed oil acted at HRCECs for 48h, inhibition of cell proliferation was significant difference compared with high glucose control group (P<0. 05). Within 48h showed concentration dependence. There was no statistical difference between the low glucose group and high glucose control group (P>0. 05). lmmunocytochemical assay showed that:50ü L/mL, 100ü L/mL, 200ü L/mL Coix seed oil acted at HRCECs 48h, the expression of VEGF decreased significantly compared with the high glucose control group ( P< 0. 05 ), and in a dose- dependent manner. However, in high glucose control group, the expression of VEGF was obvious higher than that of low glucose control group (P<0. 05).
CONCLUSlON:Coix seed oil can inhibit the HRCECs proliferation and suppress the VEGF expression in high glucose environment.

Objective: The HPLC fingerprints of water extracts and alcohol extracts of Glycyrrhiza uralensis from different producing areas were established in order to provide reference for the quality control and the extraction and processing methods. Methods: The HPLC fingerprints of water extracts and alcohol extracts of 10 batches of G. uralensis from different producing areas were established by HPLC method, and the similarity evaluation and hierarchical cluster analysis of the obtained fingerprint data were carried out. Results: The fingerprint of water extracts and ethanol extracts of G. uralensis was established. There were 16 common peaks were identified from fingerprints of water extracts, and six components were identified, meanwhile, the fingerprints of water extracts showed that the similarity of 10 batches of G. uralensis was in the range of 0.030 and 0.999. Sixty common peaks were identified from fingerprints of ethanol extracts, and seven components were identified, meanwhile, the fingerprints of ethanol extracts showed that the similarity of 10 batches of G. uralensis were all greater than 0.85. The results of hierarchical cluster analysis showed that the water extracts and alcohol extracts of G. uralensis in different areas could be well classified, suggesting that natural factors such as the climate, soil conditions and other natural factors had a significant impact on the internal quality of licorice. Conclusion: A stable and reliable HPLC fingerprint evaluation method for water extracts and alcohol extracts of G uralensis has been established, which can provide a powerful theoretical basis and a guidance for the study of quality standard of G. uralensis and the optimization of extraction and processing technology, and provide a scientific basis for the choice of clinical medication.

Objective To observe the expression of axon guidance cues Slit2 and Robo4 in lung tissue of rat with acute lung injury (ALI) and explore the function of Slit2 and Robo4 in ALI.Methods Forty-eight Sprague-Dawley rats were randomly (random number) divided into control group (n =24) and ALl group (n =24).ALI model was reproduced by cecum ligation and puncture (CLP).The control group only experienced a simulated operation without CLP.Both groups were further divided into 3 subgroups with 8 rats in each subgroup:12 h,24 h,and 48 h subgroups.artery blood gas analysis,lung tissue wet/dry weight (W/D) ratio,lung histopathologic changes,pulmonary microvascular permeability were observed.The serum tumor nocrosis factor-α (TNF-α) was measured with enzyme linked immunosorbent assay (ELISA).The expression of Slit2 and Robo4 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR).The expression of Slit2 and Robo4 protein in lung tissues was assessed by immunohistochemistry.Date were analyzed by one-way ANOVA with SPSS version 13.0 software.Statistical significance was established at a P value of less than 0.05.Results Compared with the control group,in ALI rats at different time points,partial pressure of oxygen in arterial blood (PaO2) decreased significantly,lung W/D weight ratio and pulmonary microvascular permeability,the serum TNF-α increased significantly (all P ＜ 0.05),histopathology of lung revealed signs of injury.The expression of Slit2 mRNA in lung tissues was decreased markedly after CLP compared with control group [(0.56±0.13) vs.(0.87±0.05),F=41.39,P＜0.05,(0.42±0.10) vs.(0.85±0.07),F=93.54,P＜0.05,(0.26±0.08) vs.(0.89 ±0.09),F=227.05,P＜0.05].but there were no significant difference in expression of Robo4 mRNA in lung tissue between ALI group and control group [(0.86±0.07) vs.(0.83±0.05),F=0.695,P＞0.05,(0.82±0.05) vs.(0.89±0.08),F=2.061,P ＞ 0.05,(0.86 ± 0.08) vs.(0.86 ± 0.05),F =0.035,P ＞ 0.05].Immunohistochemistry study showed Slit2 protein was mainly expressed on the extracellular surface of vascular endothelial cells,while lung epithelial cell nuclei and endochylema.Robo4 protein was only expressed on the extracellular surface of vascular endothelial cells.Compared with the control group,expression of Slit2 protein in lung tissue in ALI group decreased markedly [(0.37 ± 0.05) vs.(0.45 ± 0.07),F =6.82,P ＜ 0.05,(0.32±0.06) vs.(0.47±0.09),F=23.54,P＜0.05,(0.28±0.07) vs.(0.46±0.06),F=28.01,P ＜ 0.05].As good as RT-PCR,there were no significant difference in expression of Robo4 protein in lung tissue between two groups [(0.53±0.04) vs.(0.52±0.05),F=0.155,P＞0.05,(0.53± 0.09) vs.(0.50±0.05),F=0.498,P＞0.05,(0.55±0.06) vs.(0.56±0.07),F=0.073,P ＞ 0.05].Conclusions Lung tissues of control group rats express Slit2 and Robo4.The decreased Slit2 mRNA and protein expressions in the lung tissue of rat with ALI caused by CLP may be associated with the occurrence of ALI.

Objective To evaluate the clinical efficacy and safety of treatment of non-small cell lung cancer (NSCLC) with autologous and half consistency allograft cytokines induced killer cells combined with chemotherapy. Methods We selected 42 patients with NSCLC patients as the research object. According to the group matching principle, the cases were divided into three groups: autologous CIK cells combination chemotherapy group, half consistency allograft CIK cells combination chemotherapy group, pure chemotherapy group. The autologous and allograft CIK cellular immune therapy of security, flow cytometric analysis technique (FCM) comparisons between before and after the treatment group infusion in vivo T lymphocyte subsets changes, and three treatment group clinical short-term curative effect were used in the comparison.Results FCM detection results show that CIK cell infusion after, CD+3, CD+4 / CD+8 ratio, NK cells (CD+3 CD+56)and CIK cells (CD+3 CD+56) ratio obviously higher than before treatment, autologous infusion before treatment,respectively (47.2±10.1) %, 1.0±0.1, (15.1±2.7) %, (0.7±0.2) %. After treatment respectively (58.8±12.3) %,1.3±0.2, (24.6±7.1) %, (3.8±2.2) %; Allograft infusion before treatment for (49.4±11.4) %, 0.9±0.2, (14.8±3.2) %, (0.9±0.3) % for after treatment (57.3±9.2) %, 1.4±0.3, (25.4±6.7) %, (4.3 ± 2.6) % (t = 22, 20, 19,P ＜ 0.05), and the pure chemotherapy group before and after the treatment T lymphocyte subsets level has not seen the obvious change. Clinical short-term curative effect comparison, autologous and allograft CIK cell therapy group objective efficient and disease control rates are slightly higher than the pure chemotherapy group, but the difference was not statistically significant. Respectively 21.4 %, 57.1%, and 35.7 %, 28.6 %,64.3 %, 71.4 % (x2=38.85, x2=41.24, P ＞ 0.05). Conclusion Autologous or half consistency allograft CIK cellular immune therapy is good safety and low toxicity, have certain short-term curative effect, which can effectively slowed tumor recurrence, is a worthy of popularizing clinically tumor adjuvant treatment mode.

AIM: To construct a recombinant adenovirus vector carrying AFP promoter to specifically express a targeting gene in hepatocellular carcinoma HepG2 cells. METHODS: Based on the Adeno-X~TM Expression system, the CMV promoter was replaced by a 300 bp a-fetoprotein promoter. The EGFP(enhanced green fluorescent protein) gene as a report gene was inserted to the multiple-cloning site(MCS). The normal liver LO2 cells, hepatocellular carcinoma HepG2 cells and HeLa cells were infected by the recombinant adenovirus, respectively. Northern blotting and fluorescence microscope were used to detect the transcription level of EGFP gene and its protein expression, respectively. RESULTS: Northern blotting showed that the target gene was markedly transcribed in HepG2 cells, but slightly in LO2 and HeLa cells. Under the fluorescence microscope, strong EGFP expression was seen in HepG2 cells but very weakly in HeLa and LO2 cells. CONCLUSION: Under the control of the 300 bp human AFP promoter, the target gene carried by the recombinant adenovirus was expressed in the AFP-producing HepG2 cells at a very high level, but not or very weakly in AFP negative cells. This adenovirus system can be used as a new, potent and specific approach for the gene-targeting therapy for the AFP producing primary hepatoma. [