Objective:To investigate the in vitro the anti-prostate cancer effect of a novel polypeptide,APP216,so as to provide a basis for development of new drug for treatment of prostate cancer.Methods:The polypeptide drug included the amino sequences of BH3,K237 and DG2 domain and the peptide that could be digested by PSA.The anti-prostate cancer effects of the polypeptide prodrug on prostate cancer cell line LNCaP,22RV1(secreting PSA)and PC3,DU145(secreting no PSA)were determined by MTT test and Hoechst 33258 staining.Results:MTT test revealed that the surviving rates of LNCaP and 22PV1 cells were respectively 22% and 34% 48 h after APP216(270 ?g/ml)treatment,and 9.8% and 8.2% 72 h after APP216(270 ?g/ml)treatment.The surviving rates of PC3 and DU145 cells were respectively 90% and 95% 48 h after APP216(270 ?g/ml)treatment,and 87% and 92% 72 h after APP216(270 ?g/ml)treatment.Hoechst 33258 staining showed the typical features of cell apoptosis:cell shrinkage,chromatin condensation and hypodiploid genomic DNA content in LNCaP and PC3m cells.Flow cytometry showed an apotosis rate of 36.26% 48 h after APP216(270 ?g/ml)treatment in LNCaP cells,and of 1.63% after 48 h APP216(270 ?g/ml)treatment in PC3m cells.Conclusion:APP216 has a satisfactory in vitro cytotoxicity on human PSA-secreting prostate cancer cells and can induce tumor cell apoptosis,but not on non-PSA secreting prostate cancer cells,indicating APP216 polypeptide can be specifically digested by endonuclease enzyme.Meanwhile,BH3 domain can be transferred into cells and induce apoptosis through HIV-TAT.

Objective To investigate the characteristics of CD8+ stem memory T cells (CD8+Tscm) in patients with chronic HIV-1 infection before and after antiretroviral therapy (ART) and to analyze their associations with progression of HIV-1 infection.Methods Thirty-six patients with chronic HIV-1 infection and 20 healthy subjects were enrolled in this study.Flow cytometry was performed to detect the percentages and absolute numbers of CD8+Tscm in patients with chronic HIV-1 infection before and after antiretroviral therapy (ART) as well as in healthy subjects.Correlation analysis was used to demonstrate the relationships between CD8+Tscm and markers for progression of HIV-1 infection (CD4+T cell count, HIV-1 viral load and level of activated T cells).Results The percentages and the absolute numbers of CD8+Tscm in patients with chronic HIV-1 infection had no significant change before and after ART.They were respectively positively correlated with the percentages and the absolute numbers of CD4+Tscm.The percentage of CD8+Tscm was proportional to the percentage of CD8+ central memory T cells (CD8+Tcm), but was inversely proportional to the percentage of CD8+ effector memory T cells (CD8+Tem).In addition, the percentages of CD8+Tscm in patients with HIV-1 infection were negatively correlated with the viral loads before ART.Conclusion CD8+Tscm are responsible for maintaining the homeostasis of other CD8+T cell subsets.CD8+Tscm play an important role in inhibiting viral replication.

Objective To investigate the phenotypes and the HIV-1-specific T cell responses of KIR3DL1 positive CD8 cells in patients with early HIV-1 infection. Methods Fifty-six HIV-1 antibody negative individuals and thirty-two patients with early HIV-1 infection were enrolled in the study. Fluores-cence-activated cell sorting (FACS) was performed to detect the phenotypes of KIR3DL1 receptor expressed on the surface of CD8 cells. The levels of IFN-γwere measured by intracellular cytokine staining assay after the PBMCs were stimulated with an HIV-1 Gag peptide pool. Results The percentages of KIR3DL1+CD8 T cells in HIV-1 negative individuals and patients with early HIV-1 infection were 1. 45% (0. 12%-8. 4%) and 0. 82% (0. 14%-6. 14%), respectively, and there was no significant difference between them. The percentages of KIR3DL1+CD8 Temra cells in HIV-1 negative individuals and patients with early HIV-1 infec-tion were (4. 55±3. 84)% and (6. 71±8. 50)%, respectively, which were significantly higher than the per-centages of KIR3DL1+CD8 Tem cells, which were (0. 50±0. 59)% and (1. 18±1. 39)%, respectively (all P<0. 01). Moreover, the percentages of KIR3DL1+CD8 Tem cells in patients with early HIV-1 infection were higher than those in HIV-1 negative individuals (P=0. 001 2). The percentage of KIR3DL1+CD8 Temra cells was positively correlated with the HIV-1 viral load in patients with early HIV-1 infection ( rs=0. 576,P=0. 000 9). The percentages of KIR3DL1+CD8 Temra cells in HIV-1 patients, whose viral loads were larger than 4. 0log, were much higher than those in HIV-1 patients with viral loads less than 4. 0 log (P=0. 002). Additionally, the levels of IFN-γsecreted by KIR3DL1 positive CD8 cells were much lesser than those secreted by KIR3DL1 negative CD8 cells (P<0. 000 1). Conclusion The receptor of KIR3DL1 was mainly expressed on CD8 Temra cells in both HIV-1 negative subjects and patients with early HIV-1 infec-tion. High HIV-1 viremia was associated with the high percentage of KIR3DL1+CD8 Temra cells. The KIR3DL1 positive CD8 cells induced lower HIV-1-specific T cell responses.

Objective To investigate the cytotoxicity of γδ T cells to HIV-1 latency cells in patients with early HIV-1 infection.Methods Sixteen early HIV-1-infected patients were enrolled in this study.Peripheral blood mononuclear cells (PBMCs) of patients were isolated and γδ T cells were expanded using zoledronate (5 μmol/L) and interleukin (IL)-2 (1 000 IU/mL) ex vivo.Lactic dehydrogenase (LDH) was used to detect the cytotoxic role of γδ T cells to HIV-1 latency cells(J-Lat Full Length Clonel0.6).The phenotype of γδ T cells before and after expansion and the intensity of GFP in HIV-1 latency cells were detected by flow cytometry.Results Zoledronate plus IL-2 stimulated rapid and large γδ T cells proliferation ex vivo (P＜0.001).γδ T cells showed high cytotoxici ty to latency cells,and the intensity of GFP in latency cells was decreased significantly (P＜0.05).Moreover,expanded γδ T cells displayed cytotoxic NK-like phenotype,the frequency of CD56+ Vδ2 T cells in patients with early HIV-1 infection was significantly higher than that of healthy controls.Conclusions γδ T cell has an ability to eradicate HIV-1 latency,and γδ T cell-based autologous or xenogenous adoptive immunotherapy will have promise prospects to cure HIV-1 infection.

Objective To study the targeting ability of aptamer-modified PLGA nanoparticle encapsulated with gold nanorod and liquid perflurocarbon,and observe its mediated ultrasound/photoacoustic dual-modality imaging in vitro.Methods PLGA nanoparticle was prepared using double emulsion technique.Aptamer-modified PLGA nanoparticle was obtained by connecting,then PLGA nanoparticle was conjugated with MUC1 aptamer carbodiimide technique aimed to obtain aptamer-modified PLGA nanoparticle.Targeting ability of aptamer-modified PLGA nanoparticle to MCF-7 tumor cells in vitro was tested by fluorescence microscope.Besides,there were three control groups including PLGA nanoparticle group,aptamer group and HELA cell group.A commercial photoacoustic instrument was used to detect ultrasound/photoacoustic signal.Results A significant number of aptamer-modified PLGA nanoparticles gathered around the MCF-7 cells,however,the specific binding can not be observed in control groups.Both ultrasound and photoacoustic signal intensity of the aptamer-modified PLGA nanoparticles increased significantly when excited with laser.Conclusions The aptamer-modified PLGA nanoparticle was successfully prepared,and had superior targeting ability to MCF-7 cells,Morever,ultrasound/photoacoustic signal emitted the by nanoparticle increased significantly.These properties make the targeted nanoparticle a great potential to become a distinguished uhrasound/photoacoustic dual modality contrast agent,which lays good foundation for further targeted imaging application in vivo.

AIM:To observe the effect of Wnt/β-catenin signaling pathway on diabetic ulcer.METHODS:Diabetic animal model was established in the female Wistar rats by intraperitoneal injection of low-dose streptozotocin fol-lowing high-fat diet feeding.A circular wound was made on the dorsum of the rats in both control group and diabetic group. The condition of wound healing was recorded and the structures of the wound tissues were observed by HE staining in the 2 groups at 3, 7 and 14 d after wounding.The expression ofβ-catenin, GSK-3βand Rspo-3 at mRNA and protein levels in the wound tissues was detected by RT-PCR and ELISA.RESULTS:In diabetic group, the wound healing rate was lower (P<0.05), and the inflammatory cells, fibroblast cells and new capillaries in the wound tissues were fewer than those in control group.The expression of β-catenin and Rspo-3 at mRNA and protein levels in the wound tissues in control group was significantly higher than those in diabetic group, and the expression of GSK-3βwas exactly the opposite (P<0.05). CONCLUSION:The down-regulation of Wnt/β-catenin probably resultes from the decreased level of Rspo-3, which may be one of the reasons for delaying the diabetic ulcer healing.

Novel aminopropyl-functionalized periodic mesoporous organosilicas (APMO) were successfully prepared by co-condensation of 1,2-bis (triethoxysilyl) ethane (BTSE) and 3-aminopropyltriethoxysilane (APTES) with the template of cationic surfactants cetyltrimethylammonium chlorine (C_(18)TAC) in basic medium.With the characterization measurements of powder X-ray diffraction (XRD),FT-IR and scanning electron microscopy (SEM) so on,APMO had such advantages as good monodisperse microshperes,uniformly orderd mesopore and large specific surface areas.These mesoporous materials were directly considered to be the chromatographic stationary phases and analyzed by HPLC evaluation.During the research,the as-prepared APMO column had better permeability and three polycyclic aromatic compounds realized separation.In addition,the as-prepared APMO column had certain advantages in fast analysis.

Objective To investigate the expression of NF-k B and c-myc in keratinocytes of HPV infected condyloma acuminata (CA) lesions.Methods Fifty cases of HPV 6/11 positive CA were diagnosed by in situ hybridization.The expression and distribution of NF-k B and c-myc were detected by immunohistochemistry technique (EnVision) with CA lesions and 25 normal human skins (foreskins). Results②The expression of NF-k B and c-myc was significantly higher in keratinocytes of CA lesions than those of normal controls (P＜0.01 ).NF-k B was mainly expressed in spinous layers of CA lesions, and most positive signals located significantly in the cytoplasm.C-myc was mainly expressed in the whole epidermis or in spinous and basal layers in CA lesions,and positive signals located in the nuclei.②There was a positive correlation between the expression of NF-k B and c-myc in CA lesions (r=0.89,P＜0.01).Conclusions The study demonstrates that the expression of both NF-k B and c-myc is significant- ly higher in HPV 6/11 positive CA than in normal skin.The data suggest that transcriptional factor NF-k B and c-myc may play a role in the regulation of proliferation of HPV 6/11 infected cells.

Adolescent ; Adult ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Recovery of Function ; Tibial Fractures ; diagnostic imaging ; physiopathology ; surgery ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo explore the anesthetic effect of preemptive analgesia of frequency acupoint electrical stimulation on painless-induced abortion as well as its effect on anesthetics dosage.

METHODSNinety cases of early pregnancy who selected painless-induced abortion were randomly divided into two groups, 45 cases in each group. Frequency acupoint electrical stimulation at Ciliao (BL 32) and Shenshu (BL 23), disperse-densewave, 2 Hz/100 Hz in frequency for 15 to 20 min, was applied in the group A, which was followed by intravenous anesthesia of propofol. The intravenous anesthesia of propofol was applied in the group B. The blood pressure (BP), heart rate (HR) and SpO2 before, during and after surgery, anesthetic effect and dosage, waking time and adverse events were observed in the two groups.

RESULTSThe BP and HR during and after the surgery in the group A were not statistically different from those before the surgery (all P > 0.05). The BP was reduced and HR was slowed down during the surgery in the group B, which was significantly different from those before the surgery as well as those in the group A (all P < 0.05). The dosage of propofol was (114. 3-+6. 1) mg in the group A. obviously less than (193.2 +/- 8.9) mg in the group B (P < 0.05). The waking time was (5.6 +/- 1.2) min in the group A, obviously less than (10.1 +/- 3.9) min in the group B (P < 0.05). As for anesthetic effect, the incidence of Grade I in the group A was more than the group B (P < 0.05). The adverse events, including nausea, vomiting and contractions pain in the group A were evidently less than those in the group B (all P < 0.05).

CONCLUSIONThe preemptive analgesia of frequency acupoint electrical stimulation could significantly improve anesthetic effect of painless-induced abortion, reduce dosage of anesthetics, shorten waking time of surgery and guarantee the safety of surgery.

Abortion, Induced ; Acupuncture Analgesia ; Acupuncture Points ; Adult ; Electric Stimulation ; Female ; Humans ; Pain Management ; Pregnancy ; Young Adult