BACKGROUND:To prepare glucose-responsive microcapsules which can control insulin release as changing the glucose concentration in the medium is of great significance to control the occurrence and development of diabetes mel itus. OBJECTIVE:To study the performance of glucose-responsive alginate/modified-chitosan/alginate microcapsules carryingβ-TC3 cells. METHODS:Glucose-responsive alginate/modified-chitosan/alginate microcapsules were prepared by layer-by-layer self-assembly method to evaluate the performance. And the glucose-responsive microcapsules carryingβ-TC3 cells were prepared to observe the cel proliferation within the microcapsules. RESULTS AND CONCLUSION:The integrity rate of glucose-responsive alginate/modified-chitosan/alginate microcapsules could be 95%after 48 hours oscil ation, and the hardness of microcapsules lowered, but the elasticity increased. The permeability test showed that microcapsules intercepted macromolecular substances such as bovine serum albumin and immuno-globulin G. The microcapsules could release more insulin with the increase of glucose concentration. As described above, the glucose-responsive alginate/modified-chitosan/alginate microcapsules had good mechanical strength, immunoisolation effect and glucose sensitivity. Theβ-TC3 cells entrapped in the glucose-responsive microcapsules could grow wel and the peak of cel proliferation lagged behind as compared with non-microencapsulated cells, indicating the glucose-responsive microcapsules had good biocompatibility.

Objective To establish a Loop-mediated Isothermal Amplification (LAMP) for Pseudomonas aeruginosa rapid detection. Method 152 P. aeruginosa strains isolated from nasal swabs and 30 reference strains were applied. P. aeruginosa ATCC15442 was used to develop LAMP amplification and evaluate sensitivity and specificity. Results Sensitivity of LAMP was 103 times higher than PCR, with DNA amount as 132 fg. When LAMP was applied to 30 reference strains and 152 P. aeruginosa strains , the specification was 100% while iden-tification rate reached 94.7%. Conclusion The establishment LAMP showed a promising prospect in P. aerugi-nosa rapid detection.

Objective To investigate the effect of fusidic acid cream on inflammatory reaction caused by skin barrier damage.Methods Eight male SKH-1 hairless mice were included in this study.The back of each of these mice were equally divided into six regions measuring 1 cm × 2 cm in size,which were then assigned into six groups:blank control group remaining untreated,barrier-impaired group,barrier-impaired and fusidic acid-treated group,barrier-impaired and vehicle-treated group,barrier-unimpaired and fusidic acid-treated group,barrierunimpaired and vehicle-treated group.Stratum corneum was removed by adhesive tape stripping to establish an animal model of acute skin barrier damage in the corresponding skin regions of these mice,and fusidic acid cream or vehicle was topically applied to the corresponding regions once.Twelve hours later,skin surface swab samples were collected from the back of these mice followed by bacterial culture and colony counting.Mice were then sacrificed,and skin tissue specimens were resected from these mice,and subjected to real-time fluorescence-based quantitative PCR for the measurement of the mRNA expressions of myeloid differentiation factor 88 (MyD88),interleukin-1α (IL-1α),IL-6,epidermal antibacterial peptides S100a8 and S100a9.Statistical analysis was carried out by repeated-measures analysis of variance (ANOVA) and least significant difference (LSD) test.Results The mRNA expressions of MyD88,IL-1α,IL-6,S100a8 and S100a9 were all significantly higher in the barrier-impaired group than in the blank control group (all P ＜ 0.05).Specifically,the mRNA expression level of MyD88 in the barrier-impaired group was 8 times that in the blank control group (8.3 ± 3.0 vs.0.8 ± 0.4).Compared with the barrier-impaired group,the barrier-impaired and fusidic acid-treated group showed a significant decrease in the mRNA expressions of IL-1α (2.8 ± 0.3 vs.20.1 ± 10.0,F =47.11,P ＜ 0.01),IL-6 (1.6 ± 2.3 vs.9.4 ± 4.0,F =16.18,P＜ 0.01),S100a8 (1.5 ± 1.4 vs.5.0 ± 1.6,F=59.71,P＜ 0.05) and S100a9 (1.2 ± 0.7 vs.3.4 ± 1.6,F=21.94,P ＜ 0.05).Conlusions Fusidic acid cream could attenuate the inflammatory reaction caused by acute skin barrier damage,which might partly explain its action mechanism in the treatment of inflammatory skin diseases.

Objective To evaluate the inhibitory effect of butyl flufenamate (BT) on ultraviolet (UV)-induced acute skin phototoxic reaction,and to investigate its possible mechanisms.Methods Eight SKH-1 hairless mice were included in this study.The back of each SKH-1 hairless mouse was divided into six regions,which were then randomly classified into six groups:blank group receiving no treatment,UV group receiving UV radiation only,BT + UV group and vehicle + UV group topically treated with BT ointment and vehicle respectively followed by UV radiation,UV + BT group and UV + vehicle group topically treated with BT ointment and vehicle respectively after UV radiation.Skin samples were obtained from these mice at 24 hours after treatment.Subsequently,hematoxylin-eosin (HE) staining was performed,real-time PCR was carried out to detect mRNA expressions of caspase-3,p53,COX-2,PGER1,interleukin (IL)-1β,IL-6,and an immunofluorescence assay was conducted to observe the expression of caspase-3.Statistical analysis was carried out by repeated-measures analysis of variance (ANOVA).Results Compared with the UV group,both BT + UV group and UV + BT group showed a decrease in the degree of skin edema and number of apoptotic cells at 24 hours after UV radiation.Real-time PCR showed that the mRNA expressions of caspase-3,p53,COX-2,PGER1,IL-l β and IL-6 were significantly higher in the UV group than in the blank group (all P ＜ 0.05),but significantly lower in the BT + UV group than in the UV group (all P ＜ 0.05),and only the expressions of caspase-3 and p53 mRNAs were significantly decreased in the UV + BT group compared with the UV group (both P ＜ 0.05).The immunofluorescence assay revealed that the expression of caspase-3 increased in the UV group compared with the blank group,but decreased in both BT + UV group and UV + BT group compared with the UV group.Conclusion BT could partially inhibit UV-induced acute skin phototoxicity in SKH-1 hairless mice.

This study demonstrates that nerve growth factor (NGF) plays a protective role in myocardial infarction and early reperfusion by reducing the myocardial cell apoptosis and by improving ventricular remodeling and seeks to assess the effects and mechanisms of NGF on late reperfusion after myocardial infarction. The models of late reperfusion were established by ligating the left main coronary artery and then cutting the suture 2 hours after coronary artery ligation. The rats in NGF treatment group were injected 10 µL Ad-NGF (by constructing the adenovirus vector Ad-NGF containing NGF gene) at four locations around infarction. The rats in adenoviral vector (Adv) group were injected 10 µL adenoviral cector as the NGF group. The late reperfusion group and the sham group were given normal saline as above, and the sham group underwent thracotomy without coronary ligation. On the 3rd, 7th, 14th and 28th day after operation, we investigated the role of NGF on late reperfusion by recording cardiac structure and function with echocardiography, by examining the expression of NGF and VIII factor with immunohistochemical method, and by evaluating the myocardial cell apoptosis with terminal dUTP nick end-labeling method (TUNEL). We found that the NGF group had higher expression of NGF protein (P < 0.01) and lower apoptosis index (AI) (P < 0.01 or P < 0.05) compared to the late reperfusion group and Adv group on all time points. The NGF group had remarkably higher level of neovascularization compared to the late reperfusion group on the 14th day (P < 0.01) and the 28th day (P < 0.05). The NGF group also had higher LVEF and FS levels compared to the late reperfusion group on the 14th day (P = 0.006, P = 0.006) and on the 28th day (P = 0.000, P = 0.000). Whereas the NGF group had lower LVEDD, LVESD (P = 0.038, P = 0.000) and lower LVEDV, LVESV (P = 0.001, P = 0.000) on the 28th day compared to late reperfusion group. In this experiment, the NGF gene carried by adenovirus vector had been transfected and obviously increased the expression of NGF protein in NGF group. NGF may help postpone the myocardial remodeling and improve the heart function by promoting the myocardial neovascularization and inhibiting myocardial apoptosis.
Adenoviridae ; Animals ; Apoptosis ; Disease Models, Animal ; Echocardiography ; Genetic Therapy ; Myocardial Infarction ; therapy ; Myocardium ; pathology ; Myocytes, Cardiac ; cytology ; Nerve Growth Factor ; pharmacology ; Rats ; Reperfusion Injury ; therapy

Objective To explore the action mechanism of basic fibroblast growth factor (bFGF) on vascular smooth muscle cells (VSMCs) in spontaneously hypertensive rats (SHR).Methods Ts were obtained from SHR and SD rats.The aortic VSMCs were cultured in vitro by tissue explant method.VSMCs were treated with different concentration of exogenous bFGF (0 ng/ml,20 ng/ml,40 ng/ml,60 ng/ml,80 ng/ml,100 ng/ml) for 48 h,then cell proliferation was detected by 3 (4,5 dimethylthiazol)2,5 diphenyltetrazolium bromide (MTT) colorimetric assay.VSMCs from SHR in control group were treated with bFGF (100 ng/ml) for 48h.VSMCs from SHR in treatment group were treated with bFGF (100 ng/ml) plus Proteinkinase C(PKC) inhibitor (staurosporine) for 48 h.Results After treatment with different concentration (0 ng/ml,20 ng/ml,40 ng/ml,60 ng/ml,80 ng/ml,100 ng/ml) of bFGF for 48 h,the values measured by MTT colorimetric method were 0.402 ± 0.103,0.605 ±0.090,0.696 ± 0.131,0.812 ± 0.080,0.901 ± 0.065,1.056±0.078 respectively in aortic VSMCs from SD rats,and 0.404±0.065,0.507±0.078,0.608±0.057,0.704 ± 0.107,0.812 ± 0.097,0.908 ± 0.032 respectively in aortic VSMCs from SHR.Compared with control group,the values measured by MTT colorimetric method were decreased in treatment group (P＜0.05).The proliferative effect of bFGF in aortic VSMCs from SHR was attenuated after administration of PKC inhibitor staurosporine.Conclusions Exogenous bFGF administration promotes VSMCs proliferation in SHR and SD rats in a concentration-dependent manner.PKC plays an important role in the signal transduction mechanism in VSMCs proliferation by exogenous bFGF.

Objective To evaluate the relationship between myocardial perfusion and diastolic function with velocity vector imaging (VVI) combined with myocardial contrast echocardiography (MCE) in dog models of coronary artery stenosis at rest and stress. Methods Different stenoses in anterior descending branch were made in 8 dogs. Before and after coronary artery stenosis, VVI evaluation was made on short axis image, then MCE were performed in the left ventricular mastoid muscle section at rest and in the peak dose of dobutamine. The myocardial blood flow A·β value and peak diastolic strain rate (SR_(dia)) on the direction of the circumference of the short view were measured, and the relationship between them was analyzed. Results At rest, no significant difference of A·β value nor SR_(dia) was found between the stenotic bed and normal bed when coronary stenosis was mild or moderate. However, A·β value and SR_(dia) of the stenotic bed were smaller than those in the normal bed when coronary stenosis was severe (P<0.05). At dobutamine stress, A·β value and SR_(dia) of the stenotic bed were already less than those in the normal bed when coronary stenosis was mild or moderate. A·β values and SR_(dia) of the stenotic bed decreased further compared to the normal bed (P<0.05) when coronary artery was severe. At both rest and stress, the standard A·β value was strongly correlated with SR_(dia) (r_(rest)=0.57,r_(stress)=0.72,P<0.01). Conclusion VVI can not only evaluate the diastolic function of myocardial segments on the short axis view, but also reflect changes of myocardial perfusion to a certain extent.

Objective To study the effect of combination of Hydrochlorothiaside (HCTZ) with Captopril or spironolactone on serum high-sensitivity C-reactive protein (hsCRP) in hypertension patients. Methods A multi-centre, random and parallel control study was applied in this study. Slight and moderate hypertension patiens were se-lected. The patients were treated with placebo for two weeks and HCTZ 12.5 mga day for 6 weeks,wbo were then randomly divided into HCTZ group(12.5 mg once a day), spironolactone group(HCTZ 12.5 mg once a day + Spi-ronolactone 20 rag once a day) and captopril group(HCTZ 12.5 mg once a day + Captopril 25 rag twice a day) . By the end of one-year follow up, HCTZ group was randomly added to Spironolactone group and Captopril group because combination therapy was superior to single medication,which was recognized. During the treatment, the patients were followed up once a month ,for monitoring blood pressure, and serum hsCRP level was measured every year. Follow-up last for 4 years. By the end of 4 years, the patients were divided into treatment group and control group in order to compare the changes of serum hsCRP levels. Results At the end of 4 years, the blood pressure and serum hsCRP level were significantly decreased as compared with baseline, and were statistically different from that of control group (P <0.05 or 0.01). Multi-factor analysis showed that pre-treatment systolic blood pressue and serum hsCRP level, post-treatment decrease value of systolic blood pressue and age were the major influencing factors for the de-crease of serum hsCRP level(P < 0.05 for each). Conclusion The long-term combinaion of HCTZ with Spironolactone or Captopril not only effectivley decreases blood pressure but also decreases serum hsCRP level. The decrease value of systolic blood pressure is the major factor for influencing serum hsCRP level.

Objective To evaluate the value of L-(methyl-11C)-labeled methionine positron emissions tomography (MET PET) and MRI in target volume delineation for postoperative radiotherapy for brain high grade glioma (HGG).Methods Thirty-seven patients with supratentorial HGG were included.Both MRI and MET PET scan were performed in the same treatment position for all patients.The consistency to determine residual tumor between MRI and MET PET was analyzed.Imaging data of MET PET and MRI were coregistered using the BrainLAB image fusion software.The extension of the volume with high uptake (VMET) on MET PET were compared quantitately with the enhancing area on MRI T1W gadolinium enhancement (VGd) and the hyperintensity area on MRI T2W (VT2).Results Both MET PET and MRI were positive for 19 patients and negative for 7 patients.The consistency between these two scans was 70.3%.MET PET was integrated with MRI in 30 patients with positive MET uptake.VMET were partially or entirely outside VGd in 29 patients and VT2 in 17 patients, whereas VGd and VT2 were partially or entirely outside VMET in all patients.The maximal distance from the margin of VMET to VGd was ≥ 2.0 cm in 50%patients and the corresponding distance of VMET to VT2 was ≥ 1.0 cm in 33% patients.Conclusions The differences are existing between MET PET and MRI in determination and identification of the location and extension of residual tumor for patients with HGG.The integration of MET PET and MRI can accurately delineate radiation target volume.

Objective To evaluate the effect of intensity-modulated radiation therapy(IMRT) on parotid function in nasopharyngeal carcinoma(NPC). Methods Eighty-three NPC patients received prima-ry IMRT between 2001 and 2003. Xerostomia before radiotherapy, at the end of radiotherapy, at 6-month, 1-,2-,3-,4- and 5-year after radiotherapy were investigated, respectively. The relation between xerostomia and parotid dose distribution was analyzed. Results Of all the patients,4,31,31 and 17 had stage Ⅰ,Ⅱ,Ⅲ and ⅣA disease, respectively. Sixteen patients received chemo-radiotherapy. The median followed-up time was 65 months. The 5-year local control and regional control rate were 96% and 95% ,respectively. The 5-year overall survival rate was 80%. The mild xerostomia rate at the seven time points was 42%, 51%, 71%, 77%, 58%, 38% and 26%. The corresponding moderate xerostomia rate was 52%, 53%, 21%,8%, 3%, 2% and 2%, respectively. No serious xerostomia was observed. The mean dose of the bilateral parotid glands was 34.34 Gy. Xerostomia at 6-month after radiotherapy was positively correlated with the mean dose of the parotid glands, and D50 was the independent factor in predicting the xerostomia. Parotid function was well protected when the mean dose and D50 were no more than 33 Gy and 29 Gy,respectively. Conclusions IMRT can improve the local-regional control of NPC and protect the parotid glands from radiation-induced in-jury.