Objective: To study on in vitro release of tetanus toxoid contained in polylactide microsp heres and the influence of additives on the drug release. Methods: Some parameters affecting the rate of release were asse ssed during the in vitro experiments lasted 84 days: (1)the molecular mass of the poly mers , (2)the protein loading of the microspheres, (3)the particle sizes, (4)co-enca p sulation of additives. Results: The polylactide microspheres con taining tetanus toxoid presented considerable sustained release effect. Release kinetics of the microspheres fitted in with zero order equations. The rate of release was shown to dep end on the four parameters mentioned above. Conclusion: The poly lactide microsph eres investigated may be prospective for the development of controlled release v accines.

The hypothesis of tumor stem cells believes that a small proportion of cells which have the stemness property reside in tumor. This population is the cause of tumor formation, growth, metastasis and relapse. Only the therapy targeting this population could make tumors curable. At present, most anti-cancer protocols only kill the majority of differentiated tumor cells and hardly affect the tumor stem cells. That is why most of the therapies do not achieve good results. Disrupting the pathways and niche which regulate the tumor stem cells’ self-renewal, or inducing the tumor stem cells’ differentiation, or targeting the surface markers which distinguish the tumor stem cells from normal stem cells are all probable strategies.

Arsenic trioxide albumin microspheres (As2O3-BSA-NS) were prepared by using methods of chemical cross-linking. The desirability function (DF), calculated according to the size (<1 microm) distribution, drug loading and drug trapping efficiency, was introduced as a total index for the microspheres formulation. Four factors, inculding W/O ratio, decentralization speed, BSA concentration and stirring stabilization time, were selected and arranged in an orthogonal experimental table. The release characteristic was studied by the drug release experiment in vitro. The four factors affected DF differently. Decentralization speed behaved as the maximum (P<0.01), followed by BSA concentration (P<0.05) and the W/O ratio dose (P<0.05). Stirring stabilization time did not influence DF (P>0.05). The release experiment in vitro showed that As2O3 in As2O3-BSA-NS was released more slower than pure As2O3. It was concluded that regular As2O3-BSA-NS may be prepared by the methods of chemical cross-linking, which was optimized by orthogonal experimental analysis of different factors, and the microspheres can release As2O3 slowly.
Arsenicals/*chemistry ; Cross-Linking Reagents/pharmacology ; Delayed-Action Preparations ; Drug Carriers/*chemistry ; Drug Delivery Systems ; Microspheres ; Oxides/*chemistry ; Serum Albumin, Bovine/*chemistry ; Technology, Pharmaceutical

Objective: To investigate the preparation of pulsatile release tablets, the release of the drug in vitro and the pharmacokinetics in vivo . Methods: Dil tiazem hydrochloride（DIL） was used as model drug. The pulsatile release tabl e ts were prepared by dry-coated method with carnauba wax, bee wax and hydrophil i c cellulose as coating materials. The effects of formulation and technology on t he release characteri stic of diltiazem hydrochloride was investigated. The mechanism of pulsatile rel ease of the drug was proved by erosion test. The pharmacokinetic study on four h uman subjects was done by means of HPLC measurement. Results: In vitro ， delayed-release ti me t 10 was 2.1　h, the maximum release time t rm 4.0　h and t he pulsed-releas e time t 10-90 1.7　h. In vivo ， delayed-release time t la g was 5.7　h, the p eak time 8.5　h and the pulsed-release time 2.6　h. Conclusion: The rele ase of drug from pulsatile-released tablets of diltiazem hydrochloride was in a pulsed way both in vitro and in vivo .

Objective To optimize in vitro amplification of human γδ T cells with cytokines for tumor adoptive immunotherapy.Methods On the basis of the immobilized anti-TCR γδ antibody plus IL-2 system, other γ chain receptor family cytokines, including IL-7, IL-15 and IL-21, were tested to amplify human peripheral blood γδ T cells either alone or in diversity combination.The percentage of γδ T cells was measured by flow cytometry, and the proliferation efficiency of γδ T cells was calculated.The expression of proliferation-or cytotoxicity-related molecules on γδ T cells was examined by flow cytometry in order to explore the relevant mechanisms.The cytotoxicity of γδ T cells to Daudi cells was detected by lactate dehydrogenase.Results IL-15 alone but not IL-7 or IL-21 increases the γδ T cell purity, amplification efficiency and cytotoxicity to reach comparable levels to those of IL-2.IL-2 plus IL-15 up-regulates the expression of CD69 on γδ T cells and significantly increases their amplificationefficiency (P<0.05).IL-2 plus IL-21 enhanced the cytotoxicity of γδ T cells against Daudi cells by increasing the expression of granzyme A (P<0.001).The combination of IL-2, IL-15 and IL-21 significantly improves cytotoxicity of γδ T cells but reduces their amplification efficiency.In addition, when IL-21 was applied for a short time, it also enhanced the cytotoxicity of γδ T cells (P<0.05).Conclusions The combination of IL-2 and IL-15 as well as a short time addition of IL-21 is the best cytokine recipe to amplify human peripheral blood γδ T cells in vitro with immobilized anti-TCR γδ antibody, which can increase both the proliferation efficiency and the cytotoxicity to tumor cells of γδ T cells.

Objective To investigate diagnostic approaches of cervical glandular intraepithelial neoplasia (CGIN) for improving the diagnostic levels of CGIN.Methods Clinical data of 106 cases with CGIN admitted in hospital from Jan.2008 to Dec.2010 were analyzed retrospectively.All data from preoperative thin-prep cytologic test (TCT),cervical biopsies and postoperative pathological examination of the excised cervical tissues were reviewed.Results Among 106 patients,62 cases (58.5％,62/106) were low grade CGIN (L-CGIN),44 cases (41.5％,44/106) were high grade CGIN (H-CGIN) ; 25 cases (23.6％,25/106) were pure CGIN and 81 cases (76.4％,81/106) were CGIN mixed with cervical intraepithelial neoplasia (CIN).Fifteen cases (14.2％,15/106) were found atypical glandular cell (AGC)by TCT..In the 15 cases,there were 4 cases (6.5％,4/62) L-CGIN,and 11 cases (25.0％,11/44)H-CGIN,there was significant difference between the two groups (P ＜ 0.05) ; among 15 cases with AGC,11 cases of them (44.0％,11/25) were pure CGIN,4 cases (4.9％,4/81) mixed with CIN,in which there were significant difference (P ＜0.01).Seven cases (25.0％,7/28) were detected glandular lesions in 28 cases by endocervical curettage (ECC).Totally 23 cases (22.8％,23/101) were detected CGIN by colposcopy-directed biopsy,11 cases (19.0％,11/58) were with L-CGIN,12 cases (27.9％,12/43)H-CGIN,there was no significant difference between them (P ＞ 0.05).Among the 23 cases,13 eases(52.0％,13/25) were pure CGIN,10 cases (12.3％,10/81) CGIN mixed with CIN,which showed significant difference (P ＜ 0.01).All 106 patients were treated,101 cases treated with cervical conization and 5 cases performed hysterectomy; 23 cases were diagnosed CGIN preoperation,the ratio of preoperative diagnosis was 21.7％ (23/106),83 cases (80.3％,83/106) diagnosed postoperatively.Conclusions Routine diagnostic methods of CGIN were not satisfaction,most CGIN were diagnosed after cervical resection.Cervical conization may play a very important role in diagnosis of CGIN.The positivity of TCT in H-CGIN was higher than L-CGIN.There was no different in diagnosing different CGIN grades by colposcopy-directed biopsy.The ratio of preoperative diagnosis of pure CGIN was higher than those with CGIN mixed with CIN.

OBJECTIVE: To establish a quality standard for Jieyinling lotion. METHODS: Cortex Phellodendri and Epimedium Herb were identified by TLC, and the content of Icariin was determined by HPLC. RESULTS: The TLC spots were fairly clear and a good reproducibility was obtained. The linear range of Icariin was 2.0 ～10.0?g(r=0.999 6),with average recovery at 100.1%,RSD =1.92%. CONCLUSION: The established method is reliable and accurate, and suitable for the quality control of Jieyinling lotion.

AIM To investigate the hypoglycemic effect of buccally delivered insulin solution (INS SOL) in normal rats. METHODS The hypoglycemic response was examined after buccal delivery of INS SOL co administered with various absorption enhancers. The pharmacological bioavailability (PA) was used to evaluate the absorption enhancement of INS SOL from the buccal cavity under various conditions compared with subcutaneous injection. RESULTS In the absence of enhancers, the PA was low(6 8%) after buccal delivery of INS SOL. However, the concomitant administration of sodium lauryl phosphate, sodium deoxycholate, Brij78 as well as lecithin appeared to be more effective in increasing the hypoglycemic effects of insulin. INS SOL (5 ??kg -1 ) containing 5% Brij 78 had the highest PA of 33%. CONCLUSION The use of proper absorption enhancers is useful for improving the buccal absorption of insulin.

Objective To explore the effects of pigmentary epithelium derived factor (PDEF) on the phenotypic and immunologic function of murine-derived dentritic cells(BMDCs) .Methods Mononuclear cells(MNCs) isolated from murine bone marrow were cultured in RPMI1640 medium containing rmGM-CSF and rmIL-4 for 5 d ,and were divided into five groups .MNCs were stimulated for 3 d with either 50 ,100 ,200ng/mL PEDF ,1 μg/mL LPS(positive control) or RPMI1640(negative control) .The expression of CD11c ,CD80 and CD86 on DCs surface were analyzed by the fluorescence activated cell sorting (FCM ) .The ability of PEDF-induced BMDCs to stimulated T cell maturation were determined by the CCK-8 method and the level of IL-12 in the culture supernatant was detected by ELISA .Results The PEDF-treated BMDCs expressed high levels of CD11c ,CD80 and CD86 ,enhanced the immunolog-ical activities of T lymphocyte and its secretion of IL-12 when compared with untreated DCs .Conclusion PEDF can significantly up-regulate the expression of DCs immunological labelled molecule in in vitro cultured murine and increase its immunological com-petence .

Objective To establish the method of suspension culture for stem cells from human pancreatic cancer cell line PANC1.Methods PANC1 cells were cultured in serum-free medium under floating-culture system.Tumor cell spheres were observed by optical microscope.Expression of CD133 and cell cycle were detected by flow cytometry.Cancer stem cells were induced to differentiate with 10%FBS,and expression of CK18,was evaluated with immunofluorescence microscope.Spheres cells were injected into the subcutaneous space of NOD/SCID rat and tumor formation was monitored weekly.Results PANC1 cells could form the stem cells spheres,and the rate of sphere formation was stable between 4%0 and 5%0 after 20 passages in vitro.The expression of CD133(5.91±0.7%)and proportion of G0/G1 phase cell(80.99±2.60%)was significantly increased in spheres cells compared with parental PANC1 cells(1.44±0.52%and 69.01±5.03%),and the difference was statistically significant(P＜0.05).When these spheres cells were cultured in media with serum,these cells gradually returned to the status of parental cells and expressed CK18,2×103 sphere cells injection could initiate tumor fornmtion in NOD/SCID rat.Conclusions Tumor spheres stem cellscould be generated under serum-free floating-culture system.The sphere cells possessed the capacities of self renew,difierentiation,and tumorigenic potential.