The dialysis method was employed to prepare blank and doxorubicin (DOX) loaded micelles formed by temperature- and pH- sensitive polyhistidine-co-polyDL-lactide-co-glycolide-co-polyethyleneglycol-co-polyDL-lactide-co-glycolide-co-polyhistidine (PHis-b-PLGA-b-PEG-b-PLGA-b-PHis). The critical micelle concentrations (CMC) of the copolymers were measured with Pyrene Fluorescent Probe Technique. The temperature- and pH- sensitive properties of the blank micelles solution were investigated by optical transmittance measurement. The morphology and diameter of DOX micelles were characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The entrapment rate and drug-loading rate were determined with dialysis method. The in vitro release study was further performed to examine the temperature- and pH-responsive drug release behavior from DOX-loaded micelles. The results indicated that the CMC, entrapment efficiency and drug-loaded amount of the micelles were 7.5 x 10(-3) g x L(-1), 85.2 +/- 3.1% and 10.4 +/- 4.5%, respectively. The DOX micelle was globular-shaped with a mean diameter of 91.1 +/- 15.8 nm. The transmittance of micelle solution consistently increased with the increasing temperature or decreasing pH. In comparison to the drug release profile at physiological conditions (37 degrees C, pH 7.4), the DOX-loaded micelles showed faster drug release rate at higher temperature (41 degrees C), lower pH (pH 7.0, pH 6.5, pH 5.0) or higher temperature and lower pH (41 degrees C, pH 5.0). This indicated that the micelles showed a temperature and pH-triggered drug release pattern. Base on the above results, it can be concluded that PHis-b-PLGA-b-PEG-b-PLGA-b-PHis block copolymer micelles which respond to temperature and pH stimuli are promising smart carriers for anti-tumor drugs with the advantages of temperature- and pH- triggered drug release.

Objective To investigate the effects of insulin-like growth factor-1（IGF-1）on the cell proliferation of human non-small-cell lung cancer（NSCLC） and the possible molecular mechanism.Methods MTT assay was used to examine the effects of IGF-1 （0.1,1,10,100 ng/mL）on the cell proliferation of NSCLC cell lines（A549,LK2,H460）,Flow cytometry（FCM）and Western blot to ana-lyze the cell cycles and the protein expression of S-Phase Kinase-Associated Proteins 2（Skp2）and CDC20 homolog 1（CDH1）,respectively.Results The cell proliferation of NSCLC cell lines（A549,LK2,H460）could be promoted by the IGF-1 at different concentrations and the proliferation rate peaked when the cells were treated with 1 ng/mL IGF-1.Compared with control,the percentage of the S-phase cell population was significantly increased after the treatment of IGF-I（P 〈 0.01）and the protein expression of SKP2 also increased obviously（P 〈0.05）.However,there was no change in the CDH1 protein expression（P 〉 0.05）.Conclusion IGF-1 may accelerate the cell-cycle pro-gression of NSCLC cells by negatively modulating p27 protein via the up-regulation of SKP2 protein expression.

AIM: To investigate whether viable apoptotic cells and phagocytosis of them affect the activation of T lymphocytes. METHODS: Ultraviolet irradiation was used to induce apoptotic cells in vitro and the model of phagocytosis of these cells was established. Cytokine TGF ?1 was detected by ELISA. The rate of apoptotic cells and phagocytosis of them were assessed by flow cytometry. Furthermore, flow cytometry was also employed to examine the expression of activation signs, such as CD69, CD25, CD71, of T lymphocytes under the intervention of apoptotic cells and macrophage which ingested apoptotic cells, to reflect whether the apoptotic cells and the phagocytosis of these cells could influence the activation of lymphocytes stimulated by Con A. RESULTS: Ingestion of apoptotic cells increased TGF ?1 secretion. Only the macrophages that had ingested apoptotic cells could suppress the activation of lymphocytes. The expression of the markers of lymphocytes activation such as CD69, CD25, CD71 had been restrained. These inhibition effects were abolished by monoclonal anti-TGF ?1 antibody. CONCLUSION: The macrophages that have ingested apoptotic cells inhibit expression of CD69, CD25 and CD71 of T lymphocytes stimulated by ConA. This effect is dependent on the increase in TGF ?1 secretion in local site. [

objective To observe whether the killing effect on HCC SMMC-7721 cell of the antihepatocellular carcinoma scFv reconstructed by pharmacy was enhanced or not.Methods Prokarycytic expression vector containing PET32a-RC-RNase was induced to express by IPTG.The inclusion body purified and Western-blotting was used.PC.CHOL and CHS was added in chloroform.Dry membrane was formed after chloroform was removed.RC-RNase protein solution was added to dissolute the membrane.Then pass the solution over a Sephadex G-50 column after ultrasound and filtrated to detect the encapsulation efficiency of the liposome.The solution reacted in EDC.SSNHS and MES for 30 minutes.Then add hdscFv to the solution in 4 ℃ over night.MTT method was used to detect the killing effect on HCC cell of immunoliposome RC-RNase,immunotoxin RC-RNase and liposome RC-RNase in vitro.Resuits The killing effect on HCC cell of immunoliposome RC-RNase is the best.but that of Iiposome RC-RNase is the worst.The respective JC50 are:3.28μg/ml,22.44μg/ml and 98.26μg/ml.Conclusion The anti-hepatocellular carcinoma scFv relomtructed by pharmacy can promote the killing effect on HCC cell and may have potential in the treatment of hepatocarcinoma.

Objective:To study the effect of rotational errors on the positioning accuracy (PA) and to assess whether correcting rotation in patients with head-neck tumors in radiotherapy or not.Methods:The image information of 34 patients with head-neck tumors treated at Zhongnan Hospital of Wuhan University between August 2019 and January 2020 was collected. Mega-voltage computed tomography (MVCT) images of each patient were taken before radiotherapy, and were registered with planned kilo-voltage computed tomography (KVCT) images by two registration methods. All information was divided into control group (translation only) and intervention group (translation and rotation) according to different registration methods, there were 144 fractioned registered images for each group, respectively. The position errors of the two registration methods were recorded and compared. Data were carried out with Wilcoxon signed rank test and Spearman rank correlation.Results:Translational errors of the control group and the intervention group were 0.10 (5.35) mm and 0.00 (5.78) mm in right-left direction, and there was a statistically significant difference ( Z=-2.675, P=0.007); 0.75 (2.78) mm and 0.60 (2.68) mm in superior-inferior direction, and there was a statistically significant difference ( Z=-2.819, P=0.005); 0.10 (0.90) mm and 0.20 (1.28) mm in anterio-posterior direction, and there was a statistically significant difference ( Z=-3.984, P<0.001). Rotational errors of the intervention group were -0.20 (0.60)°, 0.35 (2.00)°, 0.00 (0.98)° in pitch, roll, yaw, respectively. The distribute of 3D vector corrected frequency for two groups was positively skewed. The corrected cumulative frequency (CCF) varied with 3D vector, 3D vector was 8.0 mm, and 19 F and 16 F fractioned treatments of the control group and the intervention group were not corrected, respectively; 3D vector was between 8.0-13.5 mm, the corrected tendency of the intervention group was slower and fractioned treatment was completed later. The analytical results of Spearman rank correlation showed that rotational errors in pitch were negatively correlated with translational errors of the control group in superior-inferior direction ( r=-0.182, P=0.029) and the intervention group in anterio-osterior direction ( r=-0.484, P<0.001); rotational errors in roll were negatively correlated with translational errors of the intervention group in right-left direction ( r=-0.334, P<0.001); rotational errors in yaw which were positively correlated with translational errors of the intervention group in right-left direction ( r=0.370, P<0.001) were negatively correlated with translational errors of the control group in superior-inferior direction ( r=-0.171, P=0.040) and the same was true for the intervention group ( r=-0.203, P=0.015); total angles were positively correlated and negatively correlated with translational errors of the control group in superior-inferior direction ( r=0.246, P=0.003) and anterio-posterior direction ( r=-0.188, P=0.024), and positively correlated with 3D vector of the control group ( r=0.198, P=0.017), total angles were positively correlated with translational errors of the intervention group in superior-inferior direction ( r=0.170, P=0.041) and with 3D vector of the intervention group ( r=0.239, P=0.004); there were no correlations between rotational errors and the other translational errors (all P>0.05). Conclusion:Although the corrected rotation increases translational errors in anterio-posterior direction and 3D vector, it improves PA for head-neck tumors in radiotherapy. When rotational errors are not corrected, rotational offsets are present with corrected translation to decrease its effect on PA.

Objective To evaluate the PSD-007-mediated photodynamic effect on mouse osteosarcoma cell line LM-8, both in vitro and in vivo. Methods LM-8 cells were incubated with different concentrations of PSD-007 for 4 hours and then followed different laser irradiations. After photodynamic therapy (PDT), cell viability was measured using MTT assay and the optical density in each experiment was measured at 450 nm with a micro plate reader. The inhibition rate of cell growth was calculated. Four-week-old female C3H mice were used for implantation of LM-8 cells. When the diameter of tumor reached up to 7-8 mm, the mice were randomly divided into following groups: 1) control group, including untreated control, saline with laser irradiation, PSD-007 without laser irradiation; 2) PDT group, PSD-007 (5 and 10 mg/kg) was injected intravenously into the mice, and the tumor site was irradiated with laser light 6 hours after injection. Seven days after PDT, the size and weight of the tumors were measured. The inhibition rate of tumor was calculated, and all tumor specimens were taken for pathologic examination. After the diameter of tumor was 10-12 mm, the tumors were performed a marginal resection and subsequently followed 3 different treatments: without PDT (control), PDT with 240 J/cm2 or 360 J/cm2 laser irradiation. After 4 weeks treatment, the tumor recurrence rates were analyzed. Results MTT assay revealed that the cytotoxic effect of PDT on the LM-8 cells was positively correlated with the concentration of PSD-007 and the level of laser irradiation. When the concentration exceeded 4μg/ml, and the energy exceeded 6 J/cm2, the inhibition ratio was over 50%. No anti-tumor effect was observed in the cells treated with only laser irradiation or PSD-007 injection. Compared with the control group, the size and weight of the tumors were obviously decreased after PDT. PDT performed after marginal resection of the tumor reduced the rate of local recurrence. Conclusion PDT with PSD-007 showed cytotoxic effect on the LM-8 cells, and which performed after marginal resection of the tumor reduced the rate of local recurrence.

Objective To explore the mechanical properties of the hybrid patch made of porcine acellular small intestinal submucosa (P-SIS) and polydioxanone (PDO) and its effect on repairing the abdominal wall defects of rats.Methods The PDOSIS patch and 8SIS patch were constructed by weaving and vacuum lamination and the mechanical properties were measured.In the both sides of abdomen of 36 healthy female SD rats,a partially defect (lcm × lcm) was created by surgery and repaired with a same area of PDO-SIS patch or 8SIS patch,each 12 rats were randomly sacrificed at 2,4 and 8 weeks after surgery,respectively,and tissue regeneration was evaluated.Results The bending length and tensile strength of the PDO-SIS patch were stronger than the 8SIS patch (P＜0.05) with the increase of diameter.No acute inflammation occurred at the repaired sites of the two groups,but a moderate chronic inflammation was observed 2 weeks after surgery,and the inflammatory response reduced gradually over time,no obvious chronic inflammation was found in the 8th week after surgery,with no statistical differences between the two groups (P＞0.05).Two patch materials were degraded gradually in the repaired area,and the regenerated collagen tissues were deposited continuously,but no significant differences existed between the two groups in the collagen content and collagen arrangement (P＞0.05).Conclusion The PDO-SIS patch has better mechanical properties than the 8SIS patch,and does not cause strong immune rejection when used to repair abdominal partially defect of SD rats.

Objective To study the pinoresinol diglucoside (PDG) on gene regulation role of ESF-1 cells in collagen secretion, to reveal PDG repair mechanisms on scalded skin.Methods The cells cultured in vitro were divided into the control group, the estradiol group and the three different PDG doses groups. The concentration of the high, medium and low dose groups were 100, 10, 1μmol/L, and that of estradiol group were 10-3μmol/L. The activity of proliferation was detected by MTT. Then collagen type I (Col I), collagen typeⅢ (ColⅢ), tissue inhibitors of metalloproteinase 1 (TIMP-1), tissue inhibitors of metalloproteinase 2 (TIMP-2) and matrix metalloproteinase 1 (MMP-1) expression levels of mRNA after administration of cells were detected by RT-PCR.Results Compared with the control group, the proliferation of ESF-1 cells (0.559 ± 0.027, 0.552 ± 0.034vs. 0.489 ± 0.027,P<0.05) in the estradiol and medium-dose PDG was significantly higher. The expression level of mRNA of ColⅠ(0.958 ± 0.021, 0.929 ± 0.031, 0.916 ± 0.015vs. 0.844 ± 0.022), ColⅢ (0.783 ± 0.038, 0.918 ± 0.021, 0.855 ± 0.017vs. 0.678 ± 0.024), TIMP-1 (0.939 ± 0.025, 0.889 ± 0.036, 0.853 ±  0.015 vs. 0.780 ± 0.023), TIMP-2 (0.507 ± 0.024, 0.655 ± 0.037, 0.572 ± 0.025vs. 0.405 ± 0.062) in the estradiol, low-, medium-dose PDG groups were significantly higher than those in the control group (P<0.05 or P<0.01). Besides, the MMP-1 (0.343 ± 0.038, 0.407 ± 0.046, 0.435 ± 0.037vs.0.519 ± 0.041) mRNA expression level in the middle and low dose PDG groups significantly decrease (P<0.05 orP<0.01). Conclusions The PDG could enhance the activity of ESF-1 cell proliferation, increase the expression of related collagen and tissue inhibitor of metalloproteinases and inhibit that of matrix metalloproteinases to repair scalded skin.

Objective To construct recombinant adenovirus vector containing mouse CD40Ig fusion gene for the study of induction of donor-specific tolerance. Methods CD40Ig fusion gene was constructed by PCR overlapping technique, and was cloned into the shuttle plasmid pAdTtrack-CMV. The linearized shuttle plasmid was co-transfected into the E.coli strain BJ5183 with bone plasmid pAdeasy1, then the recombinant adenovirus plasmid was generated. The adenovirus was packaged and amplified in Cells 293. Results The recombinant virus AdCD40Ig was successfully constructed and its titer was 5?109 efu/ml. Conclusion AdCD40Ig can be used as an agent in experiment to induce donor-specific tolerance.

Object To investigate the pharmacokinetic parameters of Musk Protecting Heart Pellets with pH-dependent gradient-release (MPHP-pH) and Musk Protecting Heart Pills (MPHP). Methods The cardiac muscle nutritional blood flow in rat was measured as effective index. Results It was one-compartment model when the rat was ig MPHP. The minimal effecting dose was 0.54 mg/kg, the present half-life, the elimination half-life, medicinal effect and the peak time of effective action were 0.53, 1.21, 3.48 and 1.13 h, respectively. The absorption half-life, elimination half-life and peak time of effective dose were 0.23, 1.47 and 0.88 h, respectively. Statistical moment analysis showed that the mean residence time mean residence time (MRT) of effective action were 5.05 h for MPHP-pH and 2.33 h for MPHP, the MRT of effective dose were 7.70 h for MPHP-pH and 3.21 h for MPHP. The relative bioavailability of effective dose for MPHP-pH was 104.03%. Conclusion MPHP has the characteristics of fast absorption, fast eliminationand short effective action time. Whereas MPHP-pH has the characteristics of fast absorption, protonged and relaxed effective action compared with MPHP.