Objective:To investigate the possible roles of the negative co-stimulation molecule PD-L(ligand) in the T-cell mediated immune response against tumor.Methods:Flow cytometric analysis was used to examine murine tumor cell line L929 expresses PD-L(L1 and L2).Anti-PD-L blocking monoclonal antibodies combined with injection i.p.of irradiated L929 into C57BL/6 mice was employed to sensitize the T cells of the animals.2 weeks after sensitization,mice were sacrificed,and splenocytes were prepared to perform the examinations of T cell proliferation,cytotoxic T cells(CTL) activity,and apoptosis through in vitro coculture with L929 in the presence of blocking PD-L antibodies.Results:The expression of PD-L1 by L929 was detectable,whereas there was no expression of PD-L2 on these cells.Application of blocking antibody against PD-L1 or PD-L2 together with irradiated L929 cells in the process of in vivo sensitization as well as in vitro stimulation,it was shown that blockade of PD-L1 significantly enhanced the T cell proliferation induced by L929 cells and the CTL activity against L929,evidenced by the increased apoptosis and death rate of L929 cells.In control,blockade of PD-L2 had small effects on these terms.Conclusion:The negative co-stimulation molecule PD-L1 plays a role in the escape of tumor cells from host T-cell mediated tumor immunity.Therefore,blockade of PD-L1 has potential significance for tumor immunotherapy.

AIM: To analyze curative effect of four regulating-intestines prescriptions (Wumei Wan, Baitouweng Tang, Shenling Baizhu San and Tongxie Yao-fang) on the treatment of ulcerative colitis (UC) through summing up the previous animal experimental results.METHODS: We collected the conclusions in the papers related to four regulating-intestines prescriptions for the treatment of UC which were published recently, made comparisons from the aspectsof symptoms, physical sign, pathological change, cytokine and its protein expression, blood adhesion molecule, cell apoptosis and controlling gene and analyzed the pathogenesis of UC and action mechanism of four regulating-intestines prescriptions. Four regulating-intestines prescriptions included Wumei Wan,Baitouweng Tang, Shenling Baizhu San and Tongxie Yaofang. Wumei Wan is from Treatise on Exogenous Febrile Diseases and Synopsis of the Golden Chamber, which consists of dark plum, asarum herb, dried ginger, Chinese goldthread, Chinese angelica root, aconite root, pricklyash peel, cassia twig,ginseng (sun-dried ginseng) and bark of cork tree and has marked effect in treating colic caused by ascariasis and persistent dysentery. Baitouweng Tang, from Treatise on Exogenous Febrile Diseases, consists of medicinal herbs such as pulsatilla root, Chinese goldthread, bark of cork tree and ash bark, which has functions of clearing away the heat-evil, expelling superficial evils and relieving dysentery. In addition, it has marked effect in treating heat-type dysentery. Shenling Baizhu San, from Prescriptions of Peaceful benevolent Dispensary, consists of medicinal herbs such as pulp of lotus seed, coix seed, amomum fruit, balloon flower root, white hyacinth bean, poria, ginseng (sun-dried ginseng), glycyrrhiza, bighead atractylodes and rhizoma dioscoreae, which has the nature of replenishing qi to invigorate the spleen and eliminate wetness to arrest diarrhea and has marked effect on treating diarrhea due to the hypofunction of spleen. Tongxie Yao-fang which is from The Complete Works of Zhang Jingyue consists of four herbals of agehead atractylodes, root of herbaceous peony, dried tangerine peel and ledebouriella root and has the functions of soothing liver and invigorating spleen and stopping diarrhea, and has marked effect on treating liver sthenia and deficient spleen, borborygmus and abdominal pain and diarrhea.RFSULTS: ① Because UC is a chronic protracted dysentery with deficiency of vital energy and existing of evil energy, the vital energy will be harmed if its treatment is specialized in removing and dissolving the stagnation, evil energy will continue to exist and stagnation will continue to accumulate if its treatment is specialized in strengthening vital energy and inducing astringency. Only supporting healthy energy and expelling evil energy is the correct therapy method. This is in accordance with the main treatment of Wumei Wan ②Eliminating dampness and pathogenic heat from the blood to treat diarrhea is the main treatment method of Baitouweng Tang, and this is incompletely suitable for the treatment of UC.③Shenling Baizhu San had the effect on soothing liver and invigorating spleen and stopping diarrhea, and this is also incompletely suitable for the treatment of UC. ④ The prescription of Tongxie Yaofang is used for treating diarrhea caused by deficient spleen and liver sthenia, and spleen controlled by liver, and abnormal ascent and descent. It accords with main pathogenesis of UC, deficient spleen, excessive dampness of deficient spleen and it is weaker in invigorating the spleen in catabasis of UC than Shenling Baizhu San. Therefore, Wumei Wan has the best curative effect,Baitouweng Tang the second, Shengling Baizhu San the third and Tongxie Yaofang a little poor.CONCLUSION: UC belongs to recurrent dysentery, and its pathogenesis is in accordance with the main treatment of Wumei Wan, but not the other three prescriptions, so Wumei Wan is the most efficient prescription in treating ulcerative colitis. Diagnosis and treatment based on an overall analysis of signs and symptoms of TCM is the premise of obtaining the best curative effect. Modeling of animal tests must be consistent with the type of syndrome of the traditional Chinese medicine. Ating ulcerative colitis (UC) from aspects such as symptom, physical sign, pathological changes, adherence factor, cytokine and its protein expression, apoptosis and its controlling gene by means of modeling, which proves their functions and effects are different and their curative effect are also different due to their different ingredient though they all have the functions of treating. Results of this test show Wumei Wan has the best curative effect, Baitouweng Tang the second, Shenling Baizhu San the third and Tongxie Yaofang a little poor[1-11] Mechanism of four regulating-intestines prescription in the treatment of ulcerative colitis is discussed from the viewpoints of traditional Chinese medicine as follows.

The variable heavy chain region (VH) genes of 3 untreated patients with B-cell chronic lymphocytic leukemia (B-CLL) were cloned and analyzed. The VH family used was VH3-11, VH3-72 and VH3-33. More than 2% difference from the corresponding germline gene was detected in all the 3 obtained potential functional genes (average 16.7). Mutation pattern analysis indicated evidence of antigen selective pressure observed in 1 of 3 cases. Our findings suggested that the tumor cells originate from post-GC cells.
Amino Acid Sequence ; Gene Rearrangement ; Immunoglobulin Variable Region/*genetics ; Leukemia, B-Cell, Chronic/*genetics ; Molecular Sequence Data ; *Mutation

The relationship between bone mineral density (BMD) and Zn, Cu, Ca levels in the meal and hair of urban and rural elderly people were studied. 470 subjects above 60 years old (urban 205 and rural 265), 178 males with an average age of 65.70 +/- 3.48 and 292 females with an average age of 65.90 +/- 4.02, were inquired. The BMD and Zn, Cu, Ca levels in the meal and hair were measured. The detected BMD in urban and rural female old people was significantly lower than that of the males; The contents of Ca and Zn in the meal of the urban females were significantly lower than those of the urban males; The Ca, Zn in the meal and Zn in the hair of the rural females were significantly lower than those of rural males (P < 0.05 or 0.01). The BMD, Ca intakes, Ca and Zn in the hair of the rural old people were significantly lower than those of the urban old people (P < 0.05 or 0.01). There was a correlation between BMD with the Ca, Zn of the hair and dietary Ca, Zn, Cu or between dietary Zn with Ca, Zn in the hair and Ca, Cu intakes. The Zn, Cu and Ca levels in the meal nutrients were correlated with BMD to some degrees. Lack of Ca and Zn in the meal can cause the reduction of BMD.
Bone Density ; Cadmium/*analysis ; Copper/*analysis ; Diet Surveys ; Hair/*chemistry ; Nutritional Status ; Osteoporosis/prevention & control ; Rural Health ; Zinc/*analysis

Objectives To study the effects of pU- vascular endothelial growth factor (VEGF)- short interfering RNA (siRNA) on the formation and apoptosis of malignant melanoma in models of nude mice and its mechanism. Methods The siRNA eukaryotic expression vector for VEGF was constructed, then transfected into A375 (a human malignant melanoma cell line) by electroporation. The nude mice models of malignant melanoma were constructed. The protein expression of VEGF and factor Ⅷ related antigen (FⅧRAg) specific for vascular endothelial cells was detected by immunohistochemical technique and morphological quantitative analysis. Microvessel density (MVD) was counted based on the endothelial cells positively stained with anti-FⅧRAg antibody. The apoptosis of the neoplasms in nude mice was quantitatively determined by TdT mediated dUTP nick end labeling (TUNEL). Results Both the number and the growth speed of the neoplasm formation were lower in the experimental group than those in the controls (both P

Objective:To predict the basic physicochemical properties ,structure and function ,and linear B-cell epitopes of the capsid protein VP1 of coxsackievirus A6(CVA6).Methods: The amino acid sequence of the CVA6 VP1 was analyzed using Bioedit software and various online tools including SubLoc ,TargetP and the others from ExPASy Bioinformatics Resource Portal.Results: The CVA6 VP1 protein was a hydrophilic protein with a relative molecular weight of 33.6 kD and an isoelectric point of 7.92.This protein containsed 24 phosphorylation sites , but no signal peptide , transmembrane domains and possible fatty acylation sites.Its secondary structure was characterized by the richest random coils , and 48.52 percent of its amino acid residues exposed at the solution inter-face.Epitope prediction by Bepipred showed a number of potential B cell epitopes in the protein ,the highest antigenicity index among them located in the region of amino acids residue 155-165.Conclusion:The basic physicochemical properties ,structure and function characteristics ,and potential linear B-cell epitopes of CVA 6 VP1 were successfully predicted , which laid foundations for the further study on the protein and the preparation of vaccines and immunological diagnostic reagents for CVA 6 infection.

Objective To explore the effects of SLC concentration gradient on suppression of tumor immune escape. Methods According to different SLC concentration, there were six groups. The HLA-Ⅰexpression and apoptosis of MCF-7 were detected with FCM,and intracellular BCL-2 expression was analyzed by western blot. The production of TGF-? was detected with ELISA. Results In a certain range of concentration gradient, following SLC increase, HLA-Ⅰexpression level on MCF-7 was improved, and apoptosis was induced but BCL-2 expression was enhanced. Moreover, the secretion of TGF-? was suppressed. Conclusion SLC inhibites tumor immune escape.

Objective:To study the effect of increased expression of Fas on the killing of ? cells Methods:The level of Fas expression and apoptosis of human ? cells strain (NIT) were detected by FACS Results:There is no Fas expression on NIT But Fas expression can detected by culturing NIT with IFN ? and/or TNF ? The apoptosis of NIT induced by sFasL and anti Fas correlates with the level of Fas expression Conclusion:Fas expression induced by cytokine may represent a factor for the initiation of ? cells destruction

Objective:To explore the mechanism of anti CD4 antibodies effect on SEB induced proliferation of the peripheral blood mononuclear cells (PBMC) Methods:Using MTT method to observe anti CD4 antibodies effect on PBMC proliferation in different conditions Using morphologic, biochemistry and flow cytometer methods to detect CD4 +T cells apoptosis induced by anti CD4 antibodies Results:Both anti CD4 human/murine chimeric antibodies and murine McAbs could inhibit the SEB induced PBMC proliferation Anti CD4 chimeric antibodies could specially induced CD4 +T cells apoptosis Conclusion:Most important inhibitory effects of the anti CD4 antibody appear to direct at TCR induced early activation signals, and their inhibitory effect of anti CD4 chimeric antibodies is close related to the monocytes The further cross linking of anti CD4 antibodies is important for inducing CD4 +T cells apoptosis

In order to further investigate the effect of annexin II (Ann-II) on tissue plasminogen activator (t-PA)-dependent plasminogen (PLG) activation and its interactive mechanism, recombinant native Ann-II bound t-PA, PLG and plasmin with high affinity was examined. The flow cytometric assay showed that the ann-II expression rate was higher in the human umbilical vein endothelial cell (HUVEC) (87.65%) than in the HL-60 cells as controls (35.79%). Two irrelevant proteins, bovine serum albumin (BSA) and equine IgG (EIG) had no effect on the production of plasmin. Ann-II-mediated enhancement of t-PA-dependent PLG activation was inhibited by epsilon-aminocaproic acid or by pretreatment of Ann-II with carboxypeptidase B with the inhibitive rate being 77.8% and 77.0%, respectively. It was revealed that the effect of Ann-II on PLG activation was specific for t-PA. Urokinase didn't bind to Ann-II, demonstrating the role of receptor-related lysine residues on activation of PLG, showing that the Ann-II-PLG interaction was dependent upon carboxyl-terminal lysine residues. These findings suggest that annexin II-mediated co-assembly of t-PA and PLG may promote plasmin generation and play a key role in modulating fibrinolysis on the endothelial surface.
Annexin A2/*pharmacology ; Cells, Cultured ; Endothelium, Vascular/cytology ; Endothelium, Vascular/*metabolism ; Fibrinolysis ; Plasminogen/*metabolism ; Recombinant Proteins/pharmacology ; Tissue Plasminogen Activator/*metabolism ; Umbilical Veins/cytology