AIM: To develop a simple, convenience, and inexpensive method on primary culture model of pancreatic islets in rats for the study of anti-diabetic drugs. METHODS: The pancreases of SD rats were separated from the pancreatic duct with cold Hank' s solution and picked. Then the pancreases were cut into pieces and repeatedly digested by collagenase at 37℃ for the short durations of the experiment. The isolated islets were identified by dithizone staining and the viability was evaluated by trypan blue staining. Pancreatic islets were incubated in RPMI 1640 or DMEM for 14 - 16 h, then they were transferred to new culture plates with the same medium mentioned above. Determination of insulin content of su-pernate and cell lysate and the experiment of insulin secretion by stimulation of glucose and implantation of micewith STZ-induced diabetes were used for evaluated the function of islets. RESULTS: The viability of isolated pancreatic islets was more than 95% and the purity of cultured islets was about 85% . The insulin synthesis, secretion and sensitivity of islets stimulate by glucose which were cultured in RPMI 1640 were higher than that in DMEM. The levels of blood glucose recovered to normal in type 1 diabetic mice after islets implantation. CONCLUSION : The islets got in this study have higher purity and viability with the normal biological activity for about 7 days by this method and they can be used as a cell model for the study of diabetes in vitro .

A new teaching reform project in experiment of Human Parasitology is performed by improving experiment content and adopting a new teaching method.The project elucidates that teaching reform in experiment can enhance students'abilities in the experiment,self-teaching and problem-solving and problem-analyzing,and creative ability.

Objective:To construct the Helicobacter pylori infected C57BL/6 mice model to observe the activation of NOD1/NF-κB signaling pathways in the gastric tissues,and study its roles in inflammatory response during Hp infection.Methods:6-8 week-old C57BL/6 mice were randomly divided into two groups,the Hp infection group and the control group,and mice were given by gavage every 48 h for five times with Hp or PBS,respectively.All the animals were sacrificed at different time point and the gastric tissue were stained with hematoxylin-eosin( HE);The mRNA expression of NOD1 and RIP2 in gastric tissues were examined by RT-PCR;Levels of IFN-βand IP-10 in mice serum were assessed by ELISA;Nuclear translocation of p65 in gastric tissue was detected by Western blot.Results:Hp infection elicits an inflammatory cell response,glands in gastric tissue were reduced or atrophic,as compared with that in the control group.The levels of IP-10 and IFN-βincreased in the model group, and peaked at 16 weeks after Hp infection.Hp infection increased the mRNA expression of NOD1 and the p65 content in nuclear between 24-120 h(P<0.05),and the highest level at 48 h,subsequently the expression levels were began to decrease.The mRNA expression level of RIP2 was up-regulated after Hp was administrated, peaked at 48 h and declined after 72 h.However, the expression levels would rise again at 120 h.Conclusion: Hp infection can activate the NOD1/NF-κB signaling pathways and induce the production of IFN-βand IP-10 in gastrics of mice.

BACKGROUND:Bone marrow is the main source of mesenchymal stem cells(MSCs)at present,but its application has been limited,because of some reasons such as inconvenience of isolation,and the quantity of cells decreases with human increased age.Umbilical cord as a new source of MSCs has been widespread concerned recently.OBJECTIVE:To explore the approach of isolating and culturing MSCs from Wharton's jelly of human umbilical cord,and the methods of identifying the surface antigens and the differentiation potential.METHODS:MSCs were isolated and amplified via tissue-cultivation,and cultured by FasGrow medium.Morphology of MSCs from Wharton's jelly of human umbilical cord was observed under the optical microscope.Its immunophenotypes were detected using immunohistochemistry.The differentiation of MSCs into the osteoblasts was determined utilizing Gomori calcium-cobalt alkaline phosphatase staining,von Kossa calcium node staining,and tetracyclinefluorescence labeling.The differentiation of MSCs into the adipocytes was detected using oil red O staining.RESULTS AND CONCLUSION:MSCs were easily obtained from Wharton's jelly of human umbilical cord via the proposed approach.The primary cells grew up to 70%-80% confluence after 12-16 days of culture,and meanwhile the undifferentiated state was maintained and proliferation was stabilized after passage.The cell cycle of double increase was about 2 days,and proliferation in vitro reached twenty generation above.Surface antigen analysis showed that CD44,CD105,CD133,MHC-I were positively expressed,while CD34,CD45 were negatively expressed.Experiments of differentiation in vitro indicated that the obtained cells were capable of differentiating into fat,osteoblast and nerve-like cells.

Objective To observe the changes of tongue and pulse parameters in the patients with chronic hepatitis B(CHB)after pegylated interferon alpha-2a(PEG-IFNα-2a)treatment,to investigate its value in evaluation clinical efficacy of PEG-IFNα-2a treatment.Methods 120 patients with CHB who confirmed to the standard received PEG-IFNα-2a antiviral therapy for 48 weeks,and followed up for 24 weeks.The tongue and pulse parameters were detected by DS01-A type digital tongue and pulse presentation analyzer.The changes of liver function,serum HBV markers,HBV DNA,tongue and pulse parameters were observed before and after treatment.Results 113 patients completed the course of treatment,46 cases received complete response(response rate 40.7%).The response rate of liver stagnation and spleen deficiency group was higher than that of blood stasis group(95%CI:0.010-0.677,P <0.05).The baseline of tongue and pulse parameters had no significant difference between response group and non response group(h3 /h1,t =1.799,P =0.074;h4 /h1,t =1.383,P =0.169;h5 /h1,t′=0.461,P >0.05;W/t,t′=0.688,P >0.05;R,t =1.317,P =0.190;G,t =0.346,P =0.729;B,t =1.720,P =0.088).After 48 weeks treat-ment,and followed up for 24 weeks,the tongue and pulse parameters of response group and non response group were compared with baseline,h3 /h1,h4 /h1 decreased,R value and G value increased,the differences were statistically sig-nificant(Response group:h3 /h1,t =3.004,P =0.003;h4 /h1,t =2.702,P =0.008;R,t′=2.258,P <0.05;G,t′=3.052,P <0.05.Non response group:h3 /h1,t =1.978,P =0.049;h4 /h1,t =2.487,P =0.014;R,t′=2.661,P <0.05;G,t′=2.318,P <0.05).But there were no significant differences between the response group and no response group after treatment(h3 /h1,t′=0.191,P >0.05;h4 /h1,t =0.390,P =0.697 2;h5 /h1,t′=0.957,P >0.05;W/t,t =0.149,P =0.881;R,t =1.343,P =0.181;G,t =0.994,P =0.322;B,t =0.565,P =0.572).Conclusion The changes of tongue and pulse parameters have improved after treatment with PEG-IFNαin patients with CHB. However,the value in predicting the efficacy of antiviral therapy may be limited.

Objective To investigate correlation between Helicobacter pylori infection and incidence of myocardial infarction and cerebral infarction ,and serum homocysteine(Hcy) level .Methods A total of 72 patients with myocardial infarction ,86 patients with cerebral infarction and 80 healthy subjects were enrolled and detected for Hp‐IgG and serum Hcy by using enzyme‐linked im‐munosorbent assays and enzymatic cycling method .Relationship between Hp infection and serum level of Hcy were analyzed .Results The positive rate of Hp‐IgG and serum Hcy level in patients with myocardial infarction were significantly higher than healthy subjects(P<0 .05) .Among patients with myocardial infarction ,serum level of Hcy in Hp‐IgG positive patients was higher than that in Hp‐IgG negative patients(P<0 .05) .The positive rate of Hp‐IgG between patients with cerebral infarction and healthy subjects was without significant difference(P>0 .05) .Among patients with cerebral infarction ,serum level of Hcy between Hp‐IgG positive patients and Hp‐IgG negative patients were without significant difference(P>0 .05) .But serum Hcy level in patients with cerebral infarction was significantly higher than that in healthy subjects(P<0 .05) .Conclusion Hp infection might promote the occurrence of myocardial infarction by affecting serum level of Hcy .However ,there might be without obvious correlation between Hp infection and the occurrence of cerebral infarction .And there could be no direct association between high serum level of Hcy in patients with cerebral infarction and Hp infection .

Objective To investigate the relationship of external hydrocephl(EH) with brain-injure in the neonate.Methods By performing on the patients with brain-injure with CT and making the clinical study,the change of brain following injure and its relationship with EH were analyzed.Results Hypoxic-ischemic encephalopathy(HIE) were 36 cases,EH were 19 in the cases,6 get the cerebrospinal symptom;SAH were 22 cases,EH were 15 in the cases,3 get the cerebrospinal symptom;Intracranial hemorrhage were 8 cases,EH were 3 in the cases,3 got the cerebrospinal symptom.Conclusion All the kinds of brain-injure can get EH and the rate of EH with any brain-injure is no different.But the outcome is significantly different.

Objective To observe the therapeutic effect of arthroscopic debridement combined with oral use of Chinese medicine Guanjietong Tablets ( GT) for knee osteoarthritis in the early and middle stage. Methods Ninety knee osteoarthritis patients in the early and middle stage were evenly randomized into three groups, namely GT group, glucosamine sulfate group and blank control group. All of the three groups received arthroscopic debridement, and additionally, GT group and glucosamine sulfate group were given the corresponding medicine orally. We observed Lysholm knee scores of the three groups before operation, and 2 weeks, 3 months and one year after operation, and compared the knee joint space before operation and one year after operation showed by digital X-ray photograpy. Results ( 1) Lysholm knee scores in the three groups were increased 2 weeks, 3 months and one year after operation when compared with those before operation, the difference being significant (P<0.05) . Three months and one year after operation, the scores of GT group and glucosamine sulfate group differed from those of the blank control group (P<0.01). One year after operation, the scores of GT group differed from those of glucosamine sulfate group ( P<0.05). ( 2) The results of knee joint space showed by digital X-ray photography at knee position of weight loading indicated that the difference was insignificant between GT group and glucosamine sulfate group one year after operation ( P> 0.05) , but the difference between the two groups and the blank control group was significant (P<0.05). The difference of knee joint space was insignificant before operation and one year after operation in GT group and glucosamine sulfate group ( P>0.05) , while was significant in the blank control group ( P<0.05). Conclusion Arthroscopic debridement alone has certain short-term curative effect in treating knee osteoarthritis in the early and middle stage, but the effect will decline with the extension of time. Arthroscopic debridement combined with Guanjietong Tablets could delay the narrowing of joint space and maintain the clinical curative effect, which will delay the time for artificial joint replacement.

Animals ; Asthma ; drug therapy ; metabolism ; Genistein ; pharmacology ; therapeutic use ; Guinea Pigs ; Interleukin-4 ; genetics ; metabolism ; Janus Kinase 1 ; genetics ; metabolism ; Lung ; metabolism ; Male ; Protein Kinase Inhibitors ; pharmacology ; therapeutic use

Astragalus polysaccharides was lounded to 4-(2-aminoethylphenol), followed by labeling the APS-Tyr with fluorescein-5-isothiocyanate (FITC) at the secondary amino group. The absorption enhancement effects of low molecular weight chitosan and protamine on astragalus polysaccharides were evaluated via Caco-2 cell culture model. The results show that the fluorecent labeling compound has good stability and high sensitivity. On the other hand low molecular weight chitosan and protamine also can promoted absorption of the astragalus polysaccharides without any cytotoxity, and the absorption increase was more significant with increasing the amount of low molecular weight chitosan and protamine. At the same time, the low molecular weight chitosan has slightly better effect. The transepithelial electric resistance (TEER) of Caco-2 cells show that absorption enhancers could improve its membrane transport permeability by opening tight junctions between cells and increasing the cell membrane fluidity.
Absorption ; Astragalus Plant ; chemistry ; Biological Transport ; Caco-2 Cells ; Fluorescein-5-isothiocyanate ; chemistry ; Fluorescent Dyes ; chemistry ; Humans ; Plant Extracts ; chemistry ; pharmacokinetics ; Polysaccharides ; chemistry ; pharmacokinetics