AIMTo investigate the formation mechanism, macromolecular drug loading capacity and release property of alginate-chitosan microcapsules (ACM).

METHODSACM was prepared by emulsification-gelation method and its formation mechanism was studied by DSC analysis. Using bovine serum albumin (BSA) as model drug, the drug loading and release properties of the microcapsules on macromolecular drug were investigated.

RESULTSThe results of DSC analysis showed that there is electrostatic interaction between materials encapsulated in the microcapsule. With the increase of BSA microcapsule ratio, the BSA loading percentage rose from 9.20% to 35.08%; and with the ascent of chitosan (CTS) concentration, the BSA loading percentage increased from 30.29% to 38.12%. The BSA microcapsules whowed a two-phase release in both 0.1 mol.L-1 HCl and phosphate buffer saline (pH 7.4). With the increase of CTS concentration, the BSA release more and more slowly in 0.1 mol.L-1 HCl.

CONCLUSIONSpheric and well-dispersed alginate-chitosan microcapsules were prepared. The microcapsule showed good loading capacity to BSA as well as sustained release to a certain degree.

Alginates ; chemistry ; Biopolymers ; Calorimetry, Differential Scanning ; Capsules ; Chitin ; analogs & derivatives ; chemistry ; Chitosan ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Serum Albumin, Bovine ; administration & dosage ; Technology, Pharmaceutical ; methods

AIMTo prepare thrombus-targeted urokinase liposomes and observe its improved thrombolytic efficacy on thrombus model rats.

METHODSThe ligand H-Arg-Gly-Asp-Ser-OH (RGDS) which has specific affinity to thrombus was synthesized by liquid phase method and anchored on the surface of liposomes by incorporating its conjugate with DSPE-PEG3,500-COOH into liposomal lipid bilayers, thus thrombus-targeted liposomes were produced. Urokinase (UK) liposomes were prepared at room temperature through method modification using hydrogenated soy phosphatidylcholine (HSPC); the in vivo thrombolysis of the obtained thrombus-targeted UK liposomes and its comparison with TBS (Tris-HCl buffered solution) control, free UK and UK liposomes were assessed on common carotid artery model rats.

RESULTSThe obtained liposomes were characteristic of high UK entrapment efficiency, small mean diameter and good storage stability. At the same dose (60,000 U.kg-1), compared to the wet thrombi weights of TBS control group, those of free UK group and UK liposome group showed no statistical difference, while those of targeted UK liposomes group were significantly decreased (P < 0.001); when evaluated in term of dry thrombi weights the result was slightly different. Compared to UK liposomes of the same dose, the targeted UK liposomes showed significantly improved thrombolytic efficacy (P < 0.01 in wet weights decrease and P < 0.05 in dry weights decrease respectively).

CONCLUSIONThe targeted UK liposomes displayed good targeted thrombolytic effect.

Animals ; Disease Models, Animal ; Drug Carriers ; Drug Delivery Systems ; Fibrinolytic Agents ; administration & dosage ; therapeutic use ; Liposomes ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Technology, Pharmaceutical ; Thrombosis ; drug therapy ; Treatment Outcome ; Urokinase-Type Plasminogen Activator ; administration & dosage ; therapeutic use

AIMTo target for hepatocytic cell, liposomes was modified by special ligand.

METHODSSterically stabilized liposomes (SSL) was conjugated with asialofeticin (AF), the ligand of asialoglycoprotein receptor (ASGP-R) of hepatocyte. ASGP-R-BLM is the ASGP-R reconstructed on bilayer lipid membrane (BLM). The recognition reaction between AF-SSL and ASGP-R-BLM can be monitored by the varieties of membrane electrical parameters. The targetability of AF-SSL mediated to hepatocyte was detected by radioisotopic labeled in vitro and in vivo. The therapeutic effect of antihepatocarcinoma was observed also.

RESULTSThe lifetime of ASGP-R-BLM decreased with the added amount of AF-SSL. It was demonstrated that there was recognition reaction between AF-SSL and ASGP-R-BLM. The combination of AF-SSL with hepatocyte was significantly higher than that of SSL without AF-modified in vitro and in vivo. The survival time of rat for AF-SSL carriered ADM (adriamycin) group was much longer and the toxicities on heart, kidney and lung were lower than those SSL carried ADM group.

CONCLUSIONIt is possible to actively target the cell with specific receptor by ligand modified liposomes. The result prvide scientific basis of hepatocyte targeted liposomes.

Animals ; Antineoplastic Agents ; administration & dosage ; therapeutic use ; Asialoglycoprotein Receptor ; Asialoglycoproteins ; chemistry ; Doxorubicin ; administration & dosage ; therapeutic use ; Drug Carriers ; Drug Delivery Systems ; Fetuins ; Hepatocytes ; metabolism ; Ligands ; Lipid Bilayers ; Liposomes ; chemistry ; metabolism ; Liver ; metabolism ; Liver Neoplasms, Experimental ; drug therapy ; Male ; Mice ; Random Allocation ; Rats ; alpha-Fetoproteins ; chemistry

Objective: To investigate the effect of brain derived neurotrophic factor (BDNF) on mesenchymal stem cells (MSC) inhibiting follicular helper T cells (Tfh cells). Methods: The contents of indoleamine 2,3-dioxygenase (IDO), IL-10, TGF-β and IL-21 in MSC culture supernatant were detected by ELISA; The peripheral blood of healthy volunteers were collected, and lymphocyte in peripheral blood was separated by human lymphocyte separation solution; Co-cultures of MSC and lymphocyte were performed by Transwell chamber, and the proportion of CD4(+)CXCR5(+) Tfh cells and their subtypes were detected by flow cytometry. Results: ①The concentrations of IL-10, TGF-β, and IDO in the supernatant of BDNF group (BDNF-stimulated MSC) were higher than those of the control ones (adding PBS with the same volume) [IL-10: (42.1±4.4) ng/ml vs (19.3±2.1) ng/ml, t=4.761, P=0.009; TGF-β: (13.9±1.7) ng/ml vs (5.3±0.6) ng/ml, t=5.129, P=0.008; IDO: (441.3±56.9) ng/ml vs (226.7±37.6) ng/ml, t=3.130, P=0.035]; ②The comparisons between BDNF (co-culture of lymphocyte and BDNF-stimulated MSC) and MSC groups (co-culture of lymphocyte and MSC) were detailed as of follows: the proportion of CD4(+) CXCR5(+)Tfh cells were lower [(3.37±0.21)% vs (6.51±0.27)%, t=9.353, P<0.001], the proportion of CD4(+) CXCR5(+)CXCR3(+) CCR6(-) Tfh cells were higher [(41.14±2.04)% vs (26.72±2.57)%, t=4.383, P=0.012], CD4(+)CXCR5(+)CXCR3(-)CCR6(-) Tfh2 cells and CD4(+)CXCR5(+)CXCR3(-)CCR6(+) Tfh17 cells were lower [Tfh2: (30.16±5.38)% vs (43.26±4.11)%, t=4.426, P=0.012; Tfh17: (15.61±1.52)% vs (22.32±0.72)%, t=4.202, P=0.014], the proportion of CD4(+)CXCR5(+)Foxp3(+) Tfr cells were higher [(4.95±0.22)% vs (2.32±0.16)%, t=10.241, P<0.001], the concentration of IL-21 in the lymphocyte supernatant was lower [(0.28±0.03) ng/ml vs (0.85±0.08) ng/ml, t=6.675, P=0.003]. Conclusion: BDNF could enhance the inhibitory effect of MSC on Tfh cells through inhibiting the increasing of Tfh cells and the differentiations of Tfh2 and Tfh17 cells.
Brain-Derived Neurotrophic Factor ; Cell Differentiation ; Flow Cytometry ; Humans ; Mesenchymal Stem Cells ; T-Lymphocytes, Helper-Inducer

According to the cancer stem cell theory, cancers can be initiated by cancer stem cells. This makes cancer stem cells prime targets for therapeutic intervention. Eradicating cancer stem cells by efficient targeting agents may have the potential to cure cancer. In this review, we summarize recent breakthroughs that have improved our understanding of cancer stem cells, and we discuss the therapeutic strategy of targeting cancer stem cells, a promising future direction for cancer stem cell research.
Antineoplastic Agents ; pharmacology ; therapeutic use ; Biomarkers, Tumor ; metabolism ; Cell Differentiation ; drug effects ; Drug Carriers ; Gold ; administration & dosage ; Humans ; Nanostructures ; Neoplasms ; drug therapy ; metabolism ; pathology ; Neoplastic Stem Cells ; drug effects ; metabolism ; pathology

OBJECTIVETo investigate the clinical therapeutic results of allograft tendon for anatomical reconstruction of medial patellofemoral ligament (MPFL) for the treatment of patellar dislocations.

METHODSFrom September 2008 to June 2013, 16 patients with patellar dislocation underwent MPFL reconstructions. There were 2 males and 14 females, aged 11 to 27 years old (16 years old on average). Patellar dislocations occurred in 11 left and 5 right knees. The disease course ranged from 3 to 10 years. The frequency of dislocation ranged from 9 to 33 times (19 times on average). Affected knee joints showed patellar instability; the range of action for patella obviously increased. The X-ray films showed patellar dislocation. The preoperative Q angle was (36 ± 9)°, and the congruence angle was (63 ± 18)°. Reconstruction was performed via allograft tendon. Allograft tendon was fixed through the superomedial pole of the patella, and the other end was fixed at the natural MPFL insertion site near the medial femoral condyle with an interference screw in a bone tunnel. All the patients were evaluated postoperatively; Kujala patellofemoral scores, objective knee function, complications, and reoperations were assessed.

RESULTSPrimary healing was achieved in all cases. No infection or necrosis and absorption of grafts was observed. All the patients were followed up for an average of 16.4 months (ranged, 10 to 24 months) postoperatively. At the latest follow-up, all the patients had no pain, swelling and patellar instability; neither patella redislocation nor fracture occurred. The X-ray films showed good position of tunnel 6 months after operation, and the congruence angle was (5 ± 9)°, showing statistically significant difference when compared with preoperation (P < 0.05). The postoperative Q angle was (17 ± 8)°, the Kujala knee function score improved significantly from 45.20 ± 9.20 to 89.30 ± 6.40 at the latest follow-up, showing statistically significant difference (P < 0.05).

CONCLUSIONMPFL reconstruction improves clinical symptoms. Anatomical MPFL reconstruction is effective for patellar dislocation, and it offers good recovery of the premorbid patella mechanics. The interference screw provides firm fixation. Allograft can avoid the graft harvest site morbidity, but it increases the cost of the surgery.

Adolescent ; Adult ; Allografts ; Child ; Female ; Humans ; Ligaments, Articular ; surgery ; Male ; Patellar Dislocation ; surgery ; Patellofemoral Joint ; surgery ; Reconstructive Surgical Procedures ; methods ; Tendons ; transplantation

BACKGROUNDMicroRNAs (miRNAs) control gene expression by destabilizing target transcripts and inhibiting their translation. Aberrant expression of miRNAs has been described in many human diseases, including schizophrenia. However, the effects on miRNA expression in response to antipsychotic treatment in peripheral circulation have not been thoroughly examined.

METHODSUsing quantitative real-time PCR (qRT-PCR), We quantified the expression of seven candidate miRNAs in plasma samples of 40 first-episode schizophrenics before and after antipsychotic treatment. The patients were all treated with risperidone and achieved remission in 1 year.

RESULTSCompared with the baseline, the expression levels of miR-365 and miR-520c-3p were significantly down-regulated after 1 year of risperidone treatment (P < 0.001). There were no significant correlations between the clinical symptoms and the expression levels of these two miRNAs (P > 0.05).

CONCLUSIONSThis study analyzed possible circulating miRNAs in response to antipsychotic monotherapy for schizophrenia, the further mechanism need to be confirmed.

Adult ; Antipsychotic Agents ; therapeutic use ; Female ; Humans ; Male ; MicroRNAs ; blood ; Risperidone ; therapeutic use ; Schizophrenia ; drug therapy ; genetics

OBJECTIVETo explore the role of reversal multidrug resistance (MDR) using short hairpin RNA (shRNA) expression vectors in multidrug resistance human leukemia cell line K562/ADM.

METHODSThe oligonucleotides with 19-mer hairpin structure were synthesized. The shRNA expression vectors were constructed and introduced into K562/ADM cells. Expression of mdr1 mRNA was assessed by RT-PCR, and P-gp expression was determined by Western blot. The apoptosis and sensitivity of the K562/ADM cells to doxorubicin were quantified by flow cytometry and methyl thiazolyl tetrazolium (MTT) assays, respectively. Cellular daunorubicin accumulation was assayed by laser confocal scanning microscope (LCSM).

RESULTSIn positive clones of K562/ADM cells stably transfected with pSilencer 3.1-HI neo mdr1-A and mdr1-B shRNA expression vectors, RT-PCR showed that mdr1 mRNA expression was significantly reduced to 35.9% (P < 0.05), 27.5% (P < 0.01), respectively. Western blot showed that P-gp expression was significantly and specifically inhibited. Resistance against doxorubicin was decreased from 79-fold to 38-fold (P < 0.05), 30-fold (P < 0.01) respectively. Furthermore, the fluorescence intensity of K562/ADM cells was increased significantly compared with the control. shRNA vectors significantly enhanced the cellular daunorubicin accumulation. The percent of the apoptosis cell was significantly enhanced to 18.1% (P < 0.05) , 54.4% (P < 0.01) respectively.

CONCLUSIONSshRNA expression vectors can effectively reverse MDR, and restore the sensitivity of drug-resistance K562/ADM cells to conventional chemotherapeutic agents.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Apoptosis ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Genetic Vectors ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; Transfection

Objective To investigate the modulation effects of mesenchymal stem cells (MSC) implantation on the myofibroblasts congregating in the infarct region after myocardial infarction (MI).Methods MI was induced in SD rats by left anterior descending coronary artery ligation,and the experimental animals were assigned randomly into the sham group,MI + PBS group and MI + MSC group (myocardial injection of 0.1 ml 2 × 107/ml in four locations in the infarct region).Echocardiography,hemodynamic examinations and Masson trichrome staining were performed.Implanted MSC differentiation and myofibroblasts congregating in infarct region were investigated by immunofluorescence staining.TGF-β1-Smad2 signaling pathway was examined by real-time RT-PCR and Western blot.Results (1) Four weeks late,heart-weight/body-weight ratio [(3.04 ± 0.16) mg/g vs.(3.34 ± 0.14) mg/g,P ＜ 0.01] and myocardial infarction size [(38.72 ± 2.38) ％ vs.(46.36 ± 2.81) ％,P ＜ 0.01] were significantly reduced in MI + MSC group than in MI + PBS group,while scar thickness of infarct region was thicker [(0.93 ±0.17) mm vs.(0.65 ±0.16) mm,P=0.01],and LVEF was higher [LVEF:(32.5 ±5.9)％ vs.(26.5 ±4.5) ％,P =0.03] in MI + MSC group than in MI + PBS group.(2) Myofibroblasts congregating in the infarct region was significantly enhanced in MI + MSC group compared with MI + PBS group [(196 ± 20) cells/mm2 vs.(89 ± 25) cells/mm2,P ＜ 0.01],and part of implanted MSC expressed α-SMA +.(3) TGF-β1 expression and the phosphorylating of Smad2 in the infarct region were significantly upregulated in MI + MSC group compared with MI + PBS group (all P ＜ 0.05).Conclusions MSC could improve myocardial function and promote myofibroblasts congregating in the infarct region via activating the TGF-β1-Smad2 signaling pathway in this model.

Objective:To analyze the effects of two decomposition algorithms of dual-energy cone beam CT (DECBCT) (direct decomposition and iterative decomposition) on the image quality and material decomposition accuracy of different sizes of phantoms.Methods:Different sizes of imaging parts of patients were simulated using the combination of CatPhan604 phantoms and customized annuluses. CBCT with high energy of 140 kVp and low energy of 100 kVp were acquired using the Varian Edge CBCT system. Then the material decomposition of DECBCT images was performed using the two algorithms. The electron density (ED) and contrast-to-noise ratio (CNR) of each material in the CTP682 module were calculated. They were used to assess the decomposition accuracy and image quality of the two algorithms.Results:Based on the values in the Catphan604 manual, both algorithms have high ED accuracy. Only the ED accuracy of four materials of the smallest sized phantom showed statistical difference ( z = -4.21, 4.30, 2.87, 5.45, P < 0.05), but the average relative error was less than 1%. The CNR of the iterative decomposition algorithm was significantly higher than that of the direct decomposition, increasing by 51.8%-703.47%. The increase in the phantom size significantly reduced the accuracy of ED, and the increased amplitude of the relative error was up to a maximum of 2.52%. The large phantom size also reduced the image quality of iterative decomposition, and the decreased amplitude of CNR was up to a maximum of 39.71. Conclusions:Compared with the direct decomposition, the iterative decomposition algorithm can significantly reduce the image noise and improve the contrast without losing the accuracy of electron density in the DECBCT construction of different sizes of phantoms.