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We integrate web-based learning with team based learning,which is called WTBL method (Web-based and team-based Learning). WTBL method is constructed and applied to the teaching practice of core curriculum group courses of clinical medicine. We build some small private online courses. The students can preview online and do the case discussions by teamwork in class. The application of WTBL teaching method has realized the flipping of classroom, and helps to enhance students' self-learning ability and teamwork ability.
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Objective To investigate the effects on self-renewal of pancreatic cancer stem cells by inhibiting hedgehog signaling pathway through cyclopamine. Methods PANC1 stem cells, PANC1 adherent cells and immortalized pancreatic ductal epithelial H6C7 cells were treated with 0.5, 1, 2, 5, 10 mol/L of cyclopamine for 24, 48, 72 h. The expression of Smo mRNA and Gli1 mRNA were detected by real-time PCR.Cell growth viability was measured by CCK 8. Cell cycle and apoptosis were determined by flow cytometry.Results Seventy-two hours after cyclopamine treatment, the Smo mRNA expressions of PANC1 stem cells,PANC1 adherent cells and H6C7 cells were 1,0.83 and 2.61; the expressions of Gli mRNA were 57.27,26.35,1; the inhibitory rates were ( 37.85 ± 13.69 ) %, ( 8.53 ± 4.43 ) %, (43.55 ± 28.98 ) %. Compared with PANC1, the expressions of Smo mRNA, Gli1 mRNA and the inhibitory rate of PANC1 stem cells significantly increased ( P ＜ 0.05 ). The proportion of G1 stage of PANC1 stem cells significantly decreased from (67.41 ±6.35)% to (36.53 ±6.03)% (P ＜0.05), and the apoptosis decreased from (10.95 ±5.68) % to ( 5.73 ± 1.42 ) % ( P ＞ 0.05 ). The proportion of G1 stage of PANC1 cells significantly decreased from ( 67.64 ± 6.88 ) % to ( 53.13 ± 1.10 ) % ( P ＜ 0.05 ); the apoptosis decreased from ( 12.08 ±4.12)% to (5.66 ± 1.33)% (P ＞0.05). While both the proportion of G1 stage and apoptosis of H6C7 cells was not significantly different. Conclusions Cyclopamine can inhibit the proliferation of PANC1 stem cells via blocking hedgehog signal pathway, and the mechanism may not be associated with cell apoptosis.
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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.
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Objective To isolate and identify side population (SP) cells like cancer stem cell from human pancreatic cancer cell line SW1990, for the purpose of further evaluation of their biological characteristics. Methods Cell suspension was stained with Hoechst 33342 and PI. Then SP cells were analyzed in the fluorescence activated cell sorter. Cell growth viability was measured by MTT. Stem cell marker CD133 was determined by flow cytometry. Cloning forming efficiency was determined by cloning plating. Expression of ABCG2 protein was detected by Western blot analysis. Results The proportion of SP cells was 2.7%, however it could be completely blocked by verapamil. 9 days later, the value of A492 of SP cells was 2.1, the cloning forming efficiency was (38.7 ± 6.8) % , the positive rate of CD133 was 69.63%, which were significantly higher than cells 0. 5, ( 15.5 ± 2.8)%, 16.71% of corresponding non-SP( P <0.05). The expression of ABCG2 in SP cells was significantly higher than that in non-SP cells. Conclusions SP cells existed in human pancreatic cancer cells SW1990.
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@# 目的了解住院精神病患者并发躯体疾病的特点及解决办法。方法对住院期间并发躯体疾病的精神病患者进行调查。结果和结论住院精神病患者所患躯体疾病常涉及多个系统和器官;患者及家属对检查配合度不高,躯体疾病与精神疾病的治疗可能存在矛盾,导致住院期间并发躯体疾病精神病患者诊断、治疗困难。
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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.
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Summary: 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 μm) 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.
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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 (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.
Subject(s)
Arsenicals/chemistry , Cross-Linking Reagents/pharmacology , Delayed-Action Preparations , Drug Carriers/chemistry , Drug Delivery Systems , Microspheres , Oxides/chemistry , Serum Albumin, Bovine/chemistry , Technology, PharmaceuticalABSTRACT
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 .
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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.
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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.