Objective To study the changes of the rat's hearing threshold by long-term treatment with Vitamin-C and the reason of the changes. Methods Forty-two Wistar rats were randomly divided into two groups: the treatment group was treated with water added Vit-C and the placebo group was treated with drinking water.(1)Concentrations of malondialolehyde(MDA)of the cochlea with fluorescence chromatometry.(2) Auditory sensibilities with auditory brainstem response testing.(3) The frequency of the mtDNA~(4834)bp deletion in the cochlea with polymerase chain reaction technique. Results Hearing threshold of both groups become worse with aging,and the treatment group showed the better auditory sensitivities,the less concentrations of MDA and the less frequency of mtDNA~(4834)bp deletion compare with the placebo group. Conclusions Long-term treatment with Vitamin-C could reduce the degree of hearing loss of the rats and the concentrations of MDA of the cochlea and the frequency of mtDNA~(4834)bp deletion in the cochlea.The oxygen radicals theory should be the etiology of presbyacusis.

Aphasia is one of the major complications after stroke, which needs an effective assessment. Event-related potential (ERP) has been widely researched in neurology, the endogenous components, such as P300, N400, mismatch negativity (MMN), contingent nega-tive variation (CNV) have been widely used in diagnosis and evaluation of the impairment of brain function, including the language func-tion. This paper discussed the application of different endogenous components of ERP in aphasia after stroke, especially the comparative analysis of N400 and P300.

ObjectiveTo construct human embryonic kidney cells (HEK293) modified with human preproenkephalin (hPPE) gene.MethodshPPE gene fragments were obtained from recombinant plasmid pcDNA3.1( + )/hPPE by using restriction endonuelease Hind Ⅲ and Not Ⅰ.Homologous recombination of lentivirus and hPPE gene was produced by using recombinant DNA technology.HEK293 cells were then transfected with the recombinant lentivirus vectors.The expression of hPPE gene in HEK293 cells was detected by Western blot.ResultsThe results of DNA sequencing indicated that the positive clone of recombinant lentivirus was completely consistent with sequencing of hPPE in Genebank.The titer of the concentrated virus was 2.07 × 108 TU/ml.GFP fluorescence was not seen in HEK293 cells transfected with the lentiviral vector under fluorescence microscope.A strong fluorescence was seen in HEK293 cells transfected with Ubc-GFP-L.V.empty viral vector.Positive expression of hPPE was demonstrated in HEK293 cells transfected with lentiviral vector by Western blot.Conclusion HEK293 cells modified with hPPE gene were successfully constructed and the target gene hPPE was stably expressed in HEK293 cells.

ObjectiveTo compare the yield rate of rats islets between different collagenase digestion groups.MethodsThe SD rats were randomly divided into two groups as following by using random digits table:collagenase P group (pancreas digested by 1 mg/ml collagenase P) and type Ⅴ collagenase group (pancreas digested by 1 mg/ml type Ⅴ collagenase).After pancreas digestion,rat islet cells in two groups were culture,purified and stained with DTZ.The mean islet number and islet equivalent (IEQ) before and after purification were measured under an inverted microscope.The viability of purified islets was assessed by fluorescence staining of aridine orange (AO) and propidium iodide (PI) under the fluorescence microscopy.After purification and culture for two days,islets function was evaluated by insulin releasing tests in the two groups.ResultsBefore purification,there was no significant difference in the islets number obtained from the pancreas between two groups (P＞0.05),but there was significant difference in the IEQ (P＜0.05).After purification,the islets number in type Ⅴcollagenase group and collagenase P group was (485 ± 113)/pancrease and (643 ± 82)/pancrease,and IEQ was (674 ± 157)/pancreas and (989 ± 126)/pancreas,respectively (P＜0.05).Islet viability in type Ⅴcollagenase group and collagenase P group was (96.13 ±1.13) ％ and (96.38 ± 0.92) ％ respectively (P＞0.05).The results of insulin releasing tests revealed there was no significant difference in islet function stimulated by hypoglycemia and hyperglycemia between two groups (P＞0.05).ConclusionTwo types of collagenase are suitable for the islets digestion in rats.The stability of digestion and yield rate of purified islets in collagenase P group are higher than in type Ⅴ collagenase group.

A novel flow-injection chemiluminescence (FI-CL) method free of CL reagent was developed for the determination of captopril based on its enhancing effect on the CL derived from diperiodatoargentate(III)-sulfuric acid system. Compared with the conventional CL system, the CL system based on trivalent silver was characterized of good selectivity for the absence of CL reagent. The CL mechanism was discussed through CL spectra and UV–vis absorption spectra. The conditions of the FI-CL system were investigated and optimized. Under the optimal conditions, the relative CL intensity was linear with the captopril concentration in the range of 0.3–15.0 μg/mL. The detection limit for captopril was 0.05 μg/mL, and the relative standard deviation (n=11) was 2.0% for 5.0 μg/mL captopril. The proposed method was applied to the analysis of captopril in tablet and human urine with the recoveries of 83.1%–112.5%, and the relative standard deviations of 0.5%–4.4%. The results obtained by the proposed method agreed well with those obtained from HPLC method. The proposed method is fast, convenient, and cost-effective for the determination of captopril in medicine and biological samples.

Background:Gastric cancer is the second most frequently diagnosed cancer and the third cause of cancer death in China. Selection of candidates in high risk of gastric cancer by a simple and non-invasive marker with high sensitivity and then undergoing endoscopy is an optimal approach for large scale gastric cancer screening. Aims:To evaluate the clinical value of using trefoil factor 3(TFF3)as a serum biomarker for gastric cancer screening. Methods:Serum samples of 49 gastric cancer patients and 29 healthy subjects were collected for measurements of serum TFFs by ELISA from Jul. 2013 to Jan. 2014 at Ningbo Medical Treatment Center Lihuili Hospital. ROC curve and the area under curve(AUC)were used to verify the diagnostic performance of serum TFF1,TFF2 and TFF3 for gastric cancer. Correlations between TFFs and clinicopathological characteristics of gastric cancer were further analyzed. Results:Serum concentration of TFF3 in gastric cancer group was significantly higher than that in healthy controls[(43. 57 ± 19. 49)ng/ mL vs.(29. 97 ± 14. 20)ng/ mL, P < 0. 01],but no significant differences were found in serum concentrations of TFF1 and TFF2 between the two groups (P > 0. 05). AUC of serum TFF1,TFF2 and TFF3 for diagnosis of gastric cancer were 0. 56,0. 56 and 0. 83, respectively,which indicated that the performance of TFF3 was the best. Taken 33. 0 ng/ mL as the cut off value of TFF3, the sensitivity and specificity were 63. 3% and 82. 8% ,respectively,and the odds ratio for predicting gastric cancer was 8. 27. Significant correlations were existed between serum concentration of TFF3 and TNM stage,differentiation and lymph node metastasis of gastric cancer(P < 0. 05). Conclusions:Serum TFF3 is a promising non-invasive biomarker for gastric cancer screening.

To study the effect of glucagon-likepeptide 1(GLP-1)receptor agonist on insulin resistance and hepatic oxidative stress in rats with diabetes mellitus combined with nonalcoholic fatty liver disease. 36 male SD rats were served as the experimental animal and randomly divided into control group, model group, and GLP-1 group. The rats of control group were given routine diet with intraperitoneal injection of normal saline, those in model group were given high fat diet and intraperitoneal injection of normal saline, while GLP-1 group rats were fed with high fat diet and intraperitoneal injection of liraglutide. After 4 weeks of treatment, insulin resistance, lipid metabolism, liver injury and oxidative stress were all assessed. Serum fasting blood glucose, fasting insulin, total cholesterol, triglyceride, alanine transaminase(ALT), aspartate transaminase(AST)levels and total cholesterol, triglyceride contents in liver tissue, and as well as homeostasis model assessment for insulin resistance(HOMA-IR)levels of model group were significantly higher than those of control group, complex insulin sensitivity index(ISIcomp)level was significantly lower than that of control group; serum fasting blood glucose, fasting insulin, total cholesterol, triglyceride, ALT, AST contents and HOMA-IR levels of GLP-1 group were significantly lower than those of model group, ISIcomp level was significantly higher than that of model group; superoxide dismutase(SOD), glutathione peroxidase(GSH-Px), catalase(CAT)contents in liver tissue of model group were significantly lower, while malondialdehyde content and SOD, GSH-Px, CAT, NF-E2 related factor-2(Nrf-2), antioxidant response element(ARE), heme oxygenase-1(HO-1), quinone oxidoreductase-1(NQO-1), glutathione thiol transferase(GST)mRNA expression were significantly higher than control group; SOD, GSH-Px, CAT contents and SOD, GSH-Px, CAT, Nrf-2, ARE, HO-1, NQO-1, GST mRNA expression in the liver tissue of GLP-1 group were significantly higher, while malondialdehyde content was significantly lower than that of model group. GLP-1 receptor agonist reduces insulin resistance and liver oxidative stress injury in diabetic rats with nonalcoholic liver disease.

BACKGROUND: Lumbar spondylolysis is the common cause of teenagers' low back pain. It should apply internal fixation if conservative treatment is invalid. There are a variety of surgeries, which aims to alleviate pain and bony fusion of pars defect.OBJECTIVE: To observe clinical outcome of bone graft of pars defect plus temporary single segmental pedicle screw rod fixation for adolescent lumbar spondylolysis.METHODS: A total of 32 adolescent patients of lumbar spondylolysis were treated by bone graft of pars defect plus temporary single segmental pedicle screw rod fixation. All the patients had bilateral spondylolysis. 20 patients had no lumbar spondylolisthesis, while the others had I° spondylolisthesis. All the patients received lumbar radiograph, CT and MRI. Visual analogue scale and Oswestry disability index were utilized to evaluate pain improvement before and after operation. MacNab was used to assess efficacy. Bone graft healing at isthmus was observed with lumbar CT after fixation. The internal fixation was removed after bone fusion, then the motion of the fixed segment and the degeneration of adjacent intervertebral disc occurred were recorded.RESULTS AND CONCLUSION: (1) Completed bone fusion of pars defect was achieved in all the patients. The average period of bone union was 7.4 months. (2) The visual analogue scale and Oswestry disability index scores were significantly improved after surgery in all patients (P < 0.05). (3) The fixed segment reserved the motion after internal fixation removal. The signal of adjacent intervertebral disc of fixed segment had no changes compared to preoperative MRI. (4) Bone graft to repair isthmus defect plus temporary single segmental screw rod fixation for adolescent lumbar spondylolysis is very effective, the bone fusion is completed, and temporary fixation may effectively alleviate the degeneration of adjacent disc.

Objective To study the important virulence regulation genes of Brucella,and to understand their function.Methods Quantitative RT-PCR was used to quantify their relative transcription profiles under stress conditions and during macrophage cell infection.Results These genes were activated at different levels under these conditions and during cell infection,indicating their roles in pathogenesis at different srage of infection.Conclusion The transcription profiles of these genes have different effects about their functions.

Objective To investigate the effects of 20 Hz repetitive transcranial magnetic stimulation (rTMS) with different intensities on neurobehavior and expression of glial fibrillary acidic protein (GFAP) in ischemic penumbra of rats with cerebral infarction,so as to explore the probable mechanism. Methods Forty-three rats were randomly divided into a blank control group( n =7 ),a model control group( n =7),a sham stimulation control group(n =8) and a rTMS group (n =21) ; the rTMS group was further subdivided into 3 subgroups:80％ MT subgroup,100％ MT subgroup and 120％ MT subgroup,with 7 rats in each subgroup.The cerebral infarction model was established by right middle cerebral artery occlusion (MCAO) in each group except the blank control group.The 3 rTMS subgroups were given 14 successive blocks of 20 Hz rTMS with corresponding intensity.The sham stimulation control group received sham treatment (without any output).The model control group was given no stimulation,and the blank control group did not receive any special treatment.Functional assessments were performed at 3 different time points.After 14-day treatment,the expression of GFAP proteins in ischemic penumbra were detected by immunohistochemistry technique. Results Functional outcome reflected from 3 behavioral tests in 100％ MT subgroup after 14-day stimulation was better than 1 day after operation,while in the other rTMS subgroups functional outcomes were just better in 2 behavioral tests.The expressions of GFAP in 3 rTMS subgroups were all less than that in model control group. Conclusions The 20 Hz rTMS with 80％ MT and 100％ MT might be safe and effective to improve the functional outcome in rats with acute cerebral infarction,especially 100％ MT.Decrease of expression of GFAP in ischemic penumbra might be one of the mechanisms of beneficial effects of rTMS in ischemia brain injury.