Objective:To explore the effect of mesoporous silica (SiO 2) chitosan (CS) hydrogel loaded with neurotrophin-3 (NT-3) on repair of spinal cord injury. Methods:After 2%, 4%, and 6% NT-3/SiO 2 were dissolved in CS solution, they were added into β-glycerophosphate sodium solution for chemical cross-linking to obtain hydrogel patches of different NT-3 loadings. The specific surface area, pore size and pore volume of mesoporous SiO 2 nanoparticles were detected by specific surface area analyzer. The morphology of mesoporous SiO 2 nanoparticles and the pore structure of freeze-dried hydrogel were detected by scanning electron microscopy. Adhesion of the hydrogel was verified by spinal cord tissue. After the NT-3/SiO 2@CS hydrogel was placed in the medium, the concentrations of NT-3 were measured for 1 to 20 days. Neural stem cells (NSCs) were isolated from fetal rats and identified. Cell counting kit 8 (CCK-8) assay was used to detect the proliferation of NSCs treated with different concentrations of hydrogel. Immunofluorescence staining was used to detect the effects of NT-3/SiO 2@CS hydrogel on differentiation of NSCs. Twenty-four 8-week-old C57BL/6JNifdc female mice were randomly divided into a sham operation group (sham), a spinal cord injury group (SCI), a chitosan hydrogel group (CS) and a mesoporous SiO 2-loaded NT-3 hydrogel group (NT-3/SiO 2@CS). In the sham group, the muscle and skin were sutured immediately after laminectomy without spinal cord injury. The CS hydrogel and NT-3/SiO 2@CS hydrogel patches were implanted without treatment after spinal cord injury in the other 3 groups, respectively. Basso Mouse Scale (BMS) was used to evaluate the mice every 7 days for 8 weeks after modeling. The hot and cold board test and Catwalk gait analysis were performed at 8 weeks after surgery. Results:The mesoporous SiO 2 nanoparticles showed typical spherical morphology and a uniform particle size (about 160 nm). The specific surface area, pore volume and pore size of the mesoporous SiO 2 nanoparticles loaded with NT-3 changed from 1,039 m 2/g, 0.726 cm 3/g and 2.754 nm to 779 m 2/g, 0.403 cm 3/g and 1.903 nm, respectively. The hydrogel had a uniform internal microporous structure and good porosity so that it easily adhered to the spinal cord and achieved long-term stable release of NT-3 for at least 20 days. CCK-8 results showed that at 24 hours after the neural stem cells were laid, the cell proliferative activities in the 4%NT-3/SiO 2@CS and 6% NT-3/SiO 2@CS groups were significantly lower than that in the group untreated ( P<0.05). The immunofluorescence staining showed that the fluorescence intensity of glial fibrillary acidic protein (GFAP), the NSC marker in NT-3/SiO 2@CS, insignificantly decreased in the groups with different concentrations of NT-3/SiO 2@CS compared with the group untreated ( P>0.05). The fluorescence intensities of GFAP, MAP2 and GFAP/MAP2 in the NT-3/SiO 2@CS group were significantly higher than those in the other groups ( P<0.05). The BMS scoring for mice showed sham group>NT-3/SiO 2@CS group>SCI group and CS group for 8 weeks of modeling, and the response time of mice to cold and hot stimulations in the NT-3/SiO 2@CS group was significantly shorter than that in the SCI and CS groups. The differences above were statistically significant ( P<0.05). The Catwalk gait analysis showed that the hindlimb footprints in the NT-3/SiO 2@CS group were significantly clearer and more coherent than those in the SCI and CS groups. Conclusions:The sustained-release gel patch based on CS, SiO 2 and NT-3 has a uniform pore structure, good biocompatibility and excellent drug sustained-release effect. It can promote the differentiation of NSCs into neurons, thus contributing to recovery of motor and sensory functions after spinal cord injury.

Objective:To explore the effect of mesoporous silica (SiO 2) chitosan (CS) hydrogel loaded with neurotrophin-3 (NT-3) on repair of spinal cord injury. Methods:After 2%, 4%, and 6% NT-3/SiO 2 were dissolved in CS solution, they were added into β-glycerophosphate sodium solution for chemical cross-linking to obtain hydrogel patches of different NT-3 loadings. The specific surface area, pore size and pore volume of mesoporous SiO 2 nanoparticles were detected by specific surface area analyzer. The morphology of mesoporous SiO 2 nanoparticles and the pore structure of freeze-dried hydrogel were detected by scanning electron microscopy. Adhesion of the hydrogel was verified by spinal cord tissue. After the NT-3/SiO 2@CS hydrogel was placed in the medium, the concentrations of NT-3 were measured for 1 to 20 days. Neural stem cells (NSCs) were isolated from fetal rats and identified. Cell counting kit 8 (CCK-8) assay was used to detect the proliferation of NSCs treated with different concentrations of hydrogel. Immunofluorescence staining was used to detect the effects of NT-3/SiO 2@CS hydrogel on differentiation of NSCs. Twenty-four 8-week-old C57BL/6JNifdc female mice were randomly divided into a sham operation group (sham), a spinal cord injury group (SCI), a chitosan hydrogel group (CS) and a mesoporous SiO 2-loaded NT-3 hydrogel group (NT-3/SiO 2@CS). In the sham group, the muscle and skin were sutured immediately after laminectomy without spinal cord injury. The CS hydrogel and NT-3/SiO 2@CS hydrogel patches were implanted without treatment after spinal cord injury in the other 3 groups, respectively. Basso Mouse Scale (BMS) was used to evaluate the mice every 7 days for 8 weeks after modeling. The hot and cold board test and Catwalk gait analysis were performed at 8 weeks after surgery. Results:The mesoporous SiO 2 nanoparticles showed typical spherical morphology and a uniform particle size (about 160 nm). The specific surface area, pore volume and pore size of the mesoporous SiO 2 nanoparticles loaded with NT-3 changed from 1,039 m 2/g, 0.726 cm 3/g and 2.754 nm to 779 m 2/g, 0.403 cm 3/g and 1.903 nm, respectively. The hydrogel had a uniform internal microporous structure and good porosity so that it easily adhered to the spinal cord and achieved long-term stable release of NT-3 for at least 20 days. CCK-8 results showed that at 24 hours after the neural stem cells were laid, the cell proliferative activities in the 4%NT-3/SiO 2@CS and 6% NT-3/SiO 2@CS groups were significantly lower than that in the group untreated ( P<0.05). The immunofluorescence staining showed that the fluorescence intensity of glial fibrillary acidic protein (GFAP), the NSC marker in NT-3/SiO 2@CS, insignificantly decreased in the groups with different concentrations of NT-3/SiO 2@CS compared with the group untreated ( P>0.05). The fluorescence intensities of GFAP, MAP2 and GFAP/MAP2 in the NT-3/SiO 2@CS group were significantly higher than those in the other groups ( P<0.05). The BMS scoring for mice showed sham group>NT-3/SiO 2@CS group>SCI group and CS group for 8 weeks of modeling, and the response time of mice to cold and hot stimulations in the NT-3/SiO 2@CS group was significantly shorter than that in the SCI and CS groups. The differences above were statistically significant ( P<0.05). The Catwalk gait analysis showed that the hindlimb footprints in the NT-3/SiO 2@CS group were significantly clearer and more coherent than those in the SCI and CS groups. Conclusions:The sustained-release gel patch based on CS, SiO 2 and NT-3 has a uniform pore structure, good biocompatibility and excellent drug sustained-release effect. It can promote the differentiation of NSCs into neurons, thus contributing to recovery of motor and sensory functions after spinal cord injury.

Indocyanine green (ICG) fluorescence imaging technology has been applied in laparoscopic surgery. It is possible to highly visualize the tumor cutting edge and liver segment boundary during laparoscopic hepatectomy. Although the application of this technique in liver tumor surgery has become more and more mature, the factors affecting the quality of fluorescence imaging are still not completely clear. In this paper, we analyzed and summarized the effects of different factors such as ICG administration scheme, imaging acquisition, tumor characteristics of patients and preoperative liver function indexes on the quality of intraoperative ICG imaging, in order to provide new ideas and practical experience for clinical practice and research.

Objective The study was designed to analyze the efficacy of standardized house dust mite allergen vaccine to allergic rhinitis by subcutaneous injection and investigate the possible mechanism of specific immunotherapy(SIT).Methods From January 2011 to December 2011 a prospective study was performed in the Department of Otorhinolaryngology Head and Neck Surgery,the Affiliated Hospital of Nantong University,involving 90 patients with perennial AR,of whom 60 patients received Der p-SIT +pharmacotherapy after their approval and 30 received only pharmacotherapy.All patients were allergic to house dust mites.Symptom and medication scores were recorded three times:before the treatment,at the middle of treatment and at the end of treatment.Over a period of 1 yr.prior to and at the end of treatment,CD4 + CD25 + Foxp3 + Treg cells and Th17 cells were measured by flow cytometry.SPSS 21.0 software was used to analyze the data.Results The symptom scores using VAS and medication scores in AR patients treated with SIT and medication were reduced,the differences were significant (14.25 ± 6.40,1.00 ± 0.84 vs.32.18 ± 7.78,3.12 ± 1.54,t value was 19.65,10.71,both P ＜ 0.05).The symptom of VAS score in medication group was reduced after treatment (30.30 ±5.97 vs.20.30 ±5.79,t =10.09,P ＜0.05),but the medication score had not significant difference (P ＞ 0.05).The frequency of Thl7 cells in peripheral blood mononuclear cells were decreased in patients treated with SIT,whereas the frequency of Treg cells were increased (x2 value was 2.81,2.80,both P ＜ 0.05),but not in medication group.Conclusions Both SIT and pharmacotherapy can improve symptoms of allergic rhinitis,but SIT can also reduce medication use.The effect of immunotherapy is better than drug treatment alone.The frequency of blood Th17 cells in peripheral blood mononuclear cells were decreased in patients treated with SIT,whereas the frequency of Treg cells were increased.

Objective To investigate the role of protein kinase C (PKC) in induction of vascular endothelial growth factor (VEGF) secretion by isoflurane in primary cultured rat cardiomyocytes. Methods Primary cultured neonatal rat cardiomyocytes were randomly divided into 6 groups ( n = 6 each): control group (group C), 3 different concentration isoflurane groups (group Ⅰ1-3 ), PKC inhibitor calphostin C group (group P), and PKC inhibitor + isoflurane group (group PI). The cells were exposed to 0.7%, 1.4% and 2.1% isoflurane for6 h in group Ⅰ1-3 respectivly. Calphostin C was added to the culture medium with a final concentration of 50 nmol/L in group P. Calphostin C was added to the culture medium with a final concentration of 50 nmol/L, then the cells were exposed to 1.4% isoflurane for 6 h in group PI. VEGF concentrations and expression of PKC isoforms were determined by ELISA and Western blot respectively. Results Compared with group C, the VEGF concentration was significantly increased in group Ⅰ2 and Ⅰ3, and PKCε expression was down-regulated in the cytoplasm while upregulated in the cytomembrane in group Ⅰ2 ( P ＜ 0.01 ), but no significant change was found in the parameters mentioned above in group Ⅰ2 ( P ＞ 0.05). PKCα, PKCδ and PKCζ expression was significantly higher in the cytoplasm than in the cytomembrane in group C and Ⅰ2. VEGF concentrations were gradually increased with the increase in isoflurane concentrations ( P ＜ 0.05). VEGF concentrations were significantly lower in group PI than in Ⅰ2 ( P ＜0.05) .Conclusion Isoflurane induces VEGF secretion in primary cultured rat cardiomyocytes through translocation of PKCε from the cytoplasm to the cytomembrane, suggesting that it is a mechanism of the cardioprotective effects of isoflurane.

Objective To investigate the effects of mediastinal block(MB)on coronary atherogenesis and hemodynamics in rabbits with hyperlipoidemia.Methods Forty-eight male New Zealand white rabbits were randomly divided into 4 groups(n=12 each):control group received normal diet 150 g/d for 16 weeks,hypercholesterol group received hypercholesterol diet 150 g/d for 16 weeks,thoracic epidurial block(TEB)group received hypercholesterol diet 150 g/d for 16 weeks and TEB was performed from 13th to 16th week with 2% lidocaine 2 mg/kg twice a day,and MB group received hypercholesterol diet 150 g/d for 16 weeks and MB was performed from 13th to 16th week with 2% lidocaine 2 mg/kg twice a day.MAP was measured before and after 1st block was performed.The serum levels of total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C)and low density lipoprotein cholesterol(LDL-C)were measured on 1st day,and on4th,6th,8th and 16th week during the experiment.At the end of 16 th week,all rabbits were killed by air embolism.Heart was removed and kept in 10% formalin for a week.The ventricles were transversely sectioned at the level of papillary muscle and slices from the cross section of the ventricles were obtained for determination of the degree of atherosclerosis by microscopy.Results MAP was decreased significandy after TEB in TEB group,while there was no significant changes in MAP after MB in MB group(P＜0.05).The serum levels of TC,TG and LDL-C were significandy higher in hypercholesterol.TEB and MB groups than in control group(P＜0.05 or 0.01).The ratios of atherogenesis and intimal thickening were significantly lower in TEB and MB groups than in hyperoholesterol group(P＜0.01),there was no significant difference between TEB and MB groups.Conclusion Mediastinal block can inhibit the development of coronary atherogenesis in rabbits with hyperlipoidemia to a great degree similar to that of thoracic epidural block,but has no effect on hemodynamics.

BACKGROUND: Based on previous technique prepared for encapsulating living cells with alginate-polysine- alginate (APA) microcapsules, it has been confirmed that microencapsulated chromaffin cells have good analgesic effects. The immunoisolated effects of such microcapsule materials need to be evaluated. OBJECTIVE: This study aimed to investigate the immunological rejections of APA microencapsulated chromaffin cells transplanted into rat anterior chamber of eyes and tendon of feet, and to evaluate the immunoisolated effect of microencapsulation.DESIGN: A randomized controlled animal experiment. SETTING: Department of Anesthesiology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Forty-eight female SD rats, with the age of 3 months, were provided by the Laboratory Animal Center, Tongji Medical College, Huazhong University of Science and Technology. The protocol was carried out in accordance with ethical guidelines for the use and care of animals. Alginate and polylysine used in the experiment were the products of Sigma Company, USA. Microcapsule generator was gifted by Germany. METHODS: This study was performed at the Department of Anesthesiology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology from September 2002 to September 2003. Suprarenal medulla was taken from 6 healthy adult cadavers of brain death. After isolated, digested and cultured, suprarenal medulla was prepared into chromaffin cell suspension. Written informed consents were obtained from the family members of donors, and the protocol was given approval by the Ethics Committee of the hospital. Empty microcapsules and microencapsulated cells were prepared by APA. The 48 rats were randomly divided into the human chromaffin cell (HCC) group, the empty microcapsule group and the microencapsulated HCC (ME-HCC) group. In each group, there were two transplanted regions of anterior chamber of eyes and tendon of feet, with 8 rats used for each region. Each rat in the HCC group was perfused 2×1010 L-1 cell suspension into the anterior chamber of eyes and tendon of feet. Those in the empty microcapsule group and the ME-HCC group were perfused 100 empty capsules and ME-HCCs (100 microcapsules, 400-500 HCCs per microcapsule) into the same regions, respectively. MAIN OUTCOME MEASURES: On day 7 after transplantation, serum interleukin (IL)-2 level was determined by ELISA. Serum IgG and IgM levels were determined with a laser turbidimeter. On day 28 after transplantation, rat right eyeball and left feet were harvested, routinely sliced and stained by haematoxylin-eosin (HE). Histo-morphological structure was observed under a 40×light microscope. RESULTS: Forty-eight rats were included in the final analysis. Serum IL-2, IgG and IgM levels were significantly lower in the empty microcapsule group and ME-HCC group than in the HCC group (t=8.544-21.64, P < 0.01). A lot of lymphocyte and neutrophile infiltration could be found in the anterior chamber of eyes and tendon of feet of rats in the HCC group, but a little seen in that of the empty microcapsule group and ME-HCC group. CONCLUSION: APA microencapsulation has an effective immunoisolated effect on immunological rejection due to its good biocompatibility and mechanical stability.