Objective To study the promotive effect of chitosan liquid on wound healing.Methods Wound model was established on the back of 60 rats,which were classified into 3 groups to be treated with topical chitosan liquid (group A),basic fibroblast growth factor (group B),and sodium chloride physiological solution (group C),respectively.The time required for the healing of wound was recorded.Regenerated tissues were resected from the rats on day 3,7 and 14 after the establishment of wound model,and observed with light microscopy.Results The time required for the healing of wound was 17.3 ± 1.35 days in group A,18.2 ± 1.15 days in group B,and 24.0 ± 1.37 days in group C.For the time required for the healing of wound,no significant difference was observed between the group A and B (P ＞ 0.05),but group C significantly differed from group A and B (both P ＜ 0.05).Chitosan liquid accelerated the generation of capillary sprouts and vascular endothelial cells at the early stage of wound healing,promoted the production of fibroblasts and collagen fibers at the middle stage,and improved the organization of collagen fibers at the late stage.Conclusion Chitosan liquid exerts a promising promotive effect on wound healing.

Objective To detect the expression of survivin protein in intramedullary gliomas and evaluated the clinical significance of the survivin expression. Method Seventeen cases of intramedullary gliomas were removed by using microsurgical technique.It composed of 8 cases of ependymomas and 9 ones of astrocytomas.The patients were followed up by MRI scanning periodically.The survivin protein expression of the intramedullary gliomas were examined by immunohistochemical stain (SP). Results Total resection of the tumor was obtained in 7 cases of ependymomas and subtotal resection was undertaken in the other one case.For 9 cases of astrocytomas,total resection of the tumor achieved in 3 cases,subtoal resection in 5 cases and partial resection in one case.Survivin expression was detected in 2 samples of ependymomas and 6 ones of astrocytomas.2 astrocytoma cases were moderate positive staining,who suffering from intracranial and vertebral subarachnoid dissemination of the tumor. Conclusion The result of microsurgical treatment for intramedullary ependymomas is satisfactory.The survivin expression in astrocytoma samples is significantly higher than that in ependymoma.Moderate positive staining may correlated with subarachnoid dissemination of the tumor.

Objective To investigate the MSCT(multi-slice computed tomography) manifestation of different kinds of malignant tumor in Renal Sinus.Methods MSCT data of 31 patients with diferent kinds of malignent tumor in Renal Sinus were analyzed retrospectively.Results In our series,15 cases were renal pelvic carcinoma,and central mass in renal sinus with light or middle ehancement were revealed in their MSCT investigation,as well as pelvic filling defect and hydronephrosis to some extent were found in secretory phase of enhanced MSCT.10 cases were renal cell carcinoma with renal sinus invasion,and their MSCT muti-planar reconstruction showed the mass mainly located in renal parenchymal,with dramatical and heterogenous CT enhancement mostly,besides local oppression & latral destruction of pelvic wall were caused by renal pelvic invasion.2 cases leiomyosaocoma in renal sinus were big and had a sharp edge,after adminisration they demonstrated dramatical and heterogenous CT enhancement.3 cases were lymphnode metastasis located in renal sinus or renal gate,appeared as nodular lesion with CT enhancement,and hydronephrosis could be exsisted if renal pelvic were obstructed.1 case were retroperitoneal lymphoma,MSCT muti-planar recon-struction revealed a large retroperitoneal mass derectly invaded into renal sinus,with light or middle homogenous ehancement.Conclusion MSCT has advantages of high speed scan,excellent discrimination,and aplenty post-process technique,and is of diagonotic value to malignant tumor in renal sinus.

Using cDNA from Rehmannia glutinosa leaf as template, a 972 bp fragment of expansin gene which containing a 762 bp ORF that encoded 253 amino acids, was cloned, named RgEXPA10, which GenBank accession number for this gene is KF011918. A 1 207 bp genomic sequence of RgEXPA10 was amplified by PCR with leaf DNA as template, sequencing analysis revealed that three exons and two introns in RgEXPA10 genomic sequence, and which GenBank accession number is KF011919. Molecular and bioinformatic analyses indicated that RgEXPA10 protein have DPBB_1 and Pollen_allerg_1 domain, also including a 26 aa nuclear localization signal and a 19 aa transmembrane region. Phylogenetic analysis revealed that RgEXPA10 showed the highest homology with AtEXPA8 among the 26 α-expansins in Arabidopsis thaliana. However, the RgEXPA10 indicated the highest homology with the expansin from Solanum lycopersicum among 22 plant species. Expression patterns using qRT-PCR analysis showed that RgEXPA10 mainly expressed in unfolded leaf, followed by the tuberous root at stage of expanding period, and rarely expressed in senescing leaf. And RgEXPA10 showed higher expression level in tuberous root at 60 and 90 days after emergence. The transcription level of RgEXPA10 significantly reduced under all the three stresses including continuous cropping conditions, salinity and waterlogging. This study will lay foundations for molecular function in development and regulation of different stresses for R. glutinosa.

Objectives To investigate the occipital cortex metabolite alterations in repetitive and severe neonatal hypoglycemia rats treated with sodium pyruvate and to reveal the protective role of sodium pyruvate using high resolution 1H nuclear magnetic resonance spectroscopy.Methods Thirty-six 2-dayold Sprague-Dawley rats were randomly divided into hypoglycemia group and pyruvate group with 18 rats in each group.Rats in both groups received intraperitoneal injections of insulin (40 U/kg body weight) at 2,4 and 6 days of age to induce severe hypoglycemia (blood glucose value ≤ 1.4 mmol/L).In the hypoglycemia group,2.5 hours after insulin injection,intraperitoneal injection of 50％ glucose (2 ml/kg) was administered to terminate hypoglycemia,while in the pyruvate group,50％ glucose (2 ml/kg) and sodium pyruvate solution 2.5 ml/kg (500 mg/kg) were injected.Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay was used to observe the status of injured neurons in six neonatal rats,and metabolite changes in occipital cortex of the other 12 rats were detected by 1H nuclear magnetic resonance spectroscopy.The difference between the two groups was compared by independent-samples t test.Results Neonatal rats of both groups reached severe hypoglycemia level 2.5 hours after insulin injection.Compared with hypoglycemia group,pyruvate group had fewer injured neurons (45±5 vs 113 ± 12,t=0.782,P=0.013) and lower injured index in the occipital cortex (0.15 ± 0.03 vs 0.36 ± 0.06,t=l.143,P=0.020).Pyruvate group showed significant decreases in the concentration of taurine [(13.31 ± 2.06) vs (18.44 ± 3.86) mol/kg,t=8.231],glutamine[(1.50 ± 0.24) vs (2.02 ± 0.40) mol/kg,t=3.137],glutamate[(7.04 ± 0.95) vs (9.40 ± 1.73) mol/kg,t=6.449],aspartate[(1.51 ± 0.28) vs (2.15 ± 0.58) mol/kg,t=2.561] and creatine [(6.37±0.99) vs (8.46± 1.77) mol/kg,t =4.226] in the occipital cortex (all P'＜0.017).Conclusions Simultaneous use of glucose and sodium pyruvate to terminate hypoglycemia in repetitive and severe neonatal hypoglycemia rats can effectively alleviate severe hypoglycemia-induced occipital lobe damage via regulating excitatory amino acid neurotransmitters,energy metabolism and other metabolic pathways.

Objective: To investigate the effect of cobalt chloride (CoCl2) on transgene expression and viral particle titers in tumor cells infected by conditionally replicating adenovirus expression vector with hypoxia response elements(HRE)-regulated E1AE1B expression (Ad-5HRE-E1AE1B-RFP) and non-HRE regulated replication-deficient adenovirus expression vector (Ad-EGFP, Ad-Luc) in vitro and in vivo. Methods: Ad-5HRE-E1AE1B-RFP had five duplicated HRE and mini CMV acted as a promoter to drive E1AE1B expression and constitutive expression of RFP as reporter. The hypoxia model was optimized by exposing tumor cells to different concentrations of CoCl2. The hypoxia-inducible factor 1 alpha (HIF-1α) protein expression was determined by Western blotting. Under the optimized hypoxia model, the positive expression of exogenous gene and virus replication of Ad-5HRE-E1AE1B-RFP or Ad-EGFP-infected tumor cells were examined by conversed microscopic observation, FACS analysis and plaques formation test. Furthermore, transgene expression induced by combined application of hypoxia-inducible replicative adenovirus and replication deficient adenovirus (Ad-Luc) was also evaluated by examining the lucifererse activity in xenografted tumor models in nude mice by micro PET. Results: Western blotting results showed that CoCl2 at 0.4 and 0.08 μg/mL could stabilize and acumulate HIF-1α protein in gastric cancer SGC7901 cells, which could better mimic hypoxia condition. The microscopic observation and FACS analysis showed that CoCl2 at 0.4 μg/mL could remarkably increase the transduction efficacy of Ad-5HRE-E1AE1B-RFP, which was verified by significant increase in the percentage of positive expression of exogenous gene RFP and fluorescence intensity. But plaques formation test showed that Ad-5HRE-E1AE1B-RFP had no replication. CoCl2 0.4 μg/mL augmented the tranduction efficacy and expression levels of non-HRE regulated replication deficient adenovirus Ad-EGFP and Ad-Luc. Combined intratumoral injection of Ad-5HRE-E1AE1B-RFP and Ad-Luc significantly increased the expression of Ad-Luc in nude mice.Conclusion: CoCl2 markedly enhances transgene expression of recombinant adenovirus. However, the underlying mechanism is not only related to the CoCl2-induced hypoxia, but also probably related to regulation of gene transcription.

BACKGROUND: Genetically engineered cells have been used in the replacement therapy and gene therapy. However, how to select proper donor cells, target cells, and corresponding viral vectors is the most difficult in the therapy.OBJECTIVE: To compare the transduction efficiency of recombinant adenovirus Ad5 and AdSF35, adeno-associated virus rAAV1/2 and rAAV2, and lentivirus LV in bone marrow mesenchymal stem cells (BMSCs) and exogenous gene expression level so as to select the vectors, which can efficiently transduce BMSCs.DESIGN, TIME AND SETTING: Gene engineering controlled observation, performed in the Central Laboratory, Shanghai First People's Hospital between October 2006 and March 2007.MATERIALS: Ten Sprague Dawley rats of clean grade were used to prepare BMSCs. All recombinant viral vectors carded enhanced green fluorescent protein (EGFP) report gene. Ad5 was prepared by the Central Laboratory, Shanghai First People's Hospital. Ad5F35 was gifted by professor Qian Qi-jun from the Second Military Medical University of Chinese PLA. rAAV2 and rAAVI/2 were the products of Benyuan Zhengyang Gene Technique Co.,Ltd. LV was gifted by professor Cuo Li-be from Shanghai Institute of Biochemistry and Cytobiology, Chinese Academy of Sciences.METHODS: Rat BMSCs were in vitro isolated and cultured by density gradient centrifugation. BMSCs of passage 4 were inoculated into 24-well plate at lxlO5/well. One day later, ceils adhered to the wall and allocated to 5 groups. Ad5-EGFP [10, 100,1 000 multiplicity of infection (MOI)], Ad5F35-EGFP (10,10, 1 000 MOI), rAAVI/2-EGFP (1×104,1x10× vg), rAAV2-EGFP(1×104, 1x105vg), and LV-EGFP (30 TU) were respectively added into the 5 groups. BMSCs were transduced for 2 days with Ad virus and separately for 6 days with rAAV and LV virus.MAIN OUTCOME MEASURES: EGFP-positive expression and fluorescence intensity.RESULTS: After twenty-four hours of Ad5-EGFP transduction, EGFP-positive cells were visible under the microscope. With virus dose increasing, EGFP-positive cells increased and fluorescence intensity strengthened. Twelve days later, EGFP-positive cells gradually reduced and fluorescence intensity weakened. For Ad5F35-EGFP, its transduction was basically similar to Ad5-EGFP, but EGFP-positive cell number and fluorescence intensity were increased. After 6 days of rAAV1/2-EGFP or rAAV2-EGFP transduction, EGFP-posidve cell number and fluorescence intensity were decreased. For LV-EGFP transduction, a small amount of EGFP-positive cells could be visible on the second day, and then EGFP-positive ceils and fluorescence intensity were gradually increased until the sixth day.CONCLUSION: Ad5, Ad5F35 and LV could effectively transduce BMSCs cultured in vitro and express exogenous gene.Furthermore, transduction efficiency was correlated with virus dose in dose-dependent manner, rAAVI/2 and rAAV2 had poor/in vitro transduction efficiency.

ObjectiveTo investigate the effect of vasostatin on the migration of pancreatic cancer endothelial cells.Methods Ad-vasostatin with different concentrations of vasostatin was used to transfect pancreatic cancer endothelial cells.Ad-LacZ transfection and PBS was used as control.The effect of vasostatin gene mediated by adenovirus on the migration of pancreatic cancer endothelial cells was measured by woundhealing assay,transwell migration assay,and tube formation assay.ResultsThe scratched lines in PBS group and Ad-LacZ group were almost healed 48 hours later,while the lines in Ad-vasostatin group were rarely healed.At the MOI of 1,2,5,the migration rate of Ad-Laz group was ( 84.7 ± 2.6) ％,(80.7 ± 1.7 ) ％ and (81.3±4.0)％,while the corresponding values were (77.7 ±2.1)％,(67.3 ±2.1)％ and (38.8 ±2.1 ) ％ in Ad-vasostatin group.Transwell migration assay indicated that the number of migrated cells in Advasostatin group was inhibited in a dose-dependant manner,at the MOI of 5,the migration became significantly decreased (F=180.88,P ＜0.05).At the MOI of 1,5,10,the number of tubes in Ad-LacZ group was 118±6,120±6 and 82±5,while the corresponding values were 65±4,21±4 and 4 ±1 in Ad-vasostatin group.The number of tubes of pancreatic cancer endothelial cells was inhibited by Ad-vasostatin in a dose-dependant manner,at the MOI of 10, it was difficult to form the tubes (F-300.85,P＜0.05). Conclusions The vasostatin gene mediated by adenovirus has a significant inhibitory effect on the migation of pancreatic cancer endothelial cells in vitro in a dose-dependent manner.

Objective To investigate the clinical application and safety of DSA-guided infusion port implantation via internal jugular vein,subclavian vein and axillary vein.Methods A total of 827 patients with malignant tumors,who underwent DSA-guided intravenous infusion port implantation at the Jiangyin Municipal People's Hospital of China between March 28,2016 and June 28,2018,were enrolled in this study.According to the used approach in the port implantation,the patients were divided into internal jugular vein group(group A,n=125),subclavian vein group(group B,n=87),and axillary vein group(group C,n=615).The success rate of puncturing and the incidence of complications were compared between each other among the three groups.Results The success rates of the DSA-guided first-time puncturing in group A,B and C were 98.40％(123/125),94.25％(82/87),and 97.89％(602/615)respectively,the difference was statistically significant(P＜0.05);the success rate of puncturing in group B was lower than that in group C,and among the other groups there was no statistically significant difference(P＞0.05).All patients were followed up for a mean period of(15.56±2.91)months(range of 3-26 months).The overall incidence of postoperative early complications was 3.75％,which in group A was 4.00％(5/125),in group B was 9.20％(8/87),and in group C was 2.93％(18/615),and the incidence of complications in group B was obviously higher than that in group C(P＜0.0167).There was no statistically significant difference in the incidence of various complications between each other among the three groups(P＞0.05).The overall incidence of postoperative long-term complications was 4.72％(39/827),which in group A was 4.80％,in group B was 14.95％,and in group C was 3.25％,and the incidence of complications in group B was strikingly higher than that in group A and group C,the difference was statistically significant(P＜0.05).Comparison of groups for the rates of various complications showed that the incidence of catheter fracture in group B was higher than that in group C,and the clipping syndrome occurred only in group B,the difference was statistically significant(P＜0.0167).There was no significant difference in the incidence of other complications between each other among the three groups(P＞0.05).The abnormal port-taking rate was 1.45％,including 3 patients in group A,7 patients in group B,and 2 patients in group C.The postoperative abnormal port-taking rate in group B was the highest(8.05％),which was significantly different from that in group C(P＜0.0167).Conclusion For intravenous infusion port implantation,axillary vein approach is clinically safe,comfortable,minimally-invasive and highly-efficient method,it is superior to internal jugular vein approach and subclavian vein approach in effectively reducing the incidence of complications and improving the postoperative abnormal port-taking rate of patients.Therefore,this technique can be regarded as a first choice in clinical practice.(J Intervent Radiol,2023,32:1212-1216)