Response surface methodology (RSM) based on a three-level three-factor Box-Behnken design of experiments was used to optimize the extracellular polysaccharide content and the mycelium biomass by submerged cultivation using Pholiota squarrosa AS 5.245. The critical factors selected for the investigation were temperature, time of cultivation and volume of medium, based on the results of our previous Plackett-Burman design. The objectives of this present work were to locate optimum levels of these process parameters, and to find out interactions among them for enhancement of the yield of extracellular polysaccharide and mycelium biomass. By solving the regression equation and also by analyzing the response surface contour plots, the optimal process conditions were determined: under conditions of temperature, 28.07℃; cultivation time, 8.79 d and volume of medium, 68.51 mL, the prediction of extracellular polysaccharide content (EPC) by Pholiota squarrosa AS 5.245 was 1062.69 ?g per milliliter of fermentation liquor. While cultivation temperature, time and volume of medium were 27.60℃, 9.42 d and 54.20 mL respectively, the mycelium biomass expressed as dry cell weight (DCW) was 11.32 mg?mL -1. In order to simultaneously obtain the maximum yield of EPC and DCW, the above conditions would be located at 27.62℃, 9.19 d and 64.10 mL. In these conditions, the maximum predicted yield of EPC and DCW were found to be 1050.64 ?g?mL -1 and 11.10 mg?mL -1, respectively. These predicted values were also verified by validation experiment.

Objective To investigate the protective effect of extracellular ATP on motoneurons of spinal anterior horn from spinal root avulsion induced cell injurey. Methods Thirty-six Wister rats were randomized into 2 groups: single root avulsion and root avulsion with ATP. The left C5-C8 nerve roots were avulsed, the experiment group was gioven extracellular ATP (2 mg/kg) 0.4 ml by intraperitoneal injection and control group physiological saline water was gioven 0.4ml as the same method, all one time daily, contin-ue injection ATP or saline water 2 weeks. At 2,4 and 6 week postoperatively, C5-C8 spinal cord was har-vested after the rats were executed. The change of nitric oxide synthase (NOS) and neurofilament protein (NF-200) were observed after histochemistrical examination. Results At 2、4 and 6 week postoperatively, the survival of motorneurons in the spinal anterior was 80.48%, 73.55% and 53.43% in experiment group, was 68.90%、63.58% and 37.72% in control group;compare experiment group with control group, the le-don-induced motomeurons death in the spinal anterior bern was decreased by 11.58%、9.78% and 15.71% respectively (P < 0.01) ; the activity changes of NOS expression rate was 17.85% ,40.20% and 18.03% in experiment group, was 25.53%,53.88% and 25.58% in control group, compare experiment group with con-trol group, the activity changes of NOS expression rate was decreased by 7.68% (P < 0.01), 13.68% and 7.55% (P < 0.05) respectively. The motoneurons expressed NF-200 in the experiment group was larger than the control group, there was significant difference was found between the control group and the experiment group. Conclusion ATP might play certain protective role in survival of motoneurous after spinal root avulsion.

Objective To study the neuropsychological behavior development of preterm infants and low birth weight infants,and to provide a reference to the early prevention and intervention on developmental retardations.Methods A total of 101 preterm infants and/or low birth weight infants received the infant development test of 0 ～ 6 year-old children intelligence developmental scale for neurological development and autism behavior checklist(ABC).Results 25 boys and 5 girls suffered from different psychological mental disorders.The occurrences were as follows:10 cases with mental retardation,9 cases with the language development delay,9 cases with motor retardation,1 case with cerebral palsy and 1 case with autism spectrum disorder.The incidence of intelligence problems were that language retardation (18.9％),the fine motor (16.8％),the adapative ability (12.6％),social action (9.5 ％) and the motor delay (3.2％).There were significant differences in the scores of social communication(x2=8.88,P=0.003),adaptive ability(x2=7.41,P=0.007),the fine motor(x2 =6.22,P=0.01) and total developmental quotient(x2 =5.58,P=0.02) between city children'and rural area.The behavioral problems more consisted in self-care ability and language retardation.Conclusion Preterm infants and low birth weight infants are exposed to language,fine motor,adaptive and communication ability problems,especially the children living in country.It is necessary to improve the early education and intervention for the rural preterm infants and low birth weight infants.

The effect of Selenium(Se) on regression of atherosclerosis(AS) induced by feeding quails -with high cholesterol diet was observed. The result indicated that serum cholesterol, triglyceride and lipid peroxide significantly decreased and the ratio of high density lipoprotein cholesterol/total cholesterol increased markedly in Se treated group, that the cholesterol content of aortic tissue and AS index were significantly reduced at the end of the 15th week. It was suggested that Se had a powerful accelerated effect on regression of experimental AS.

OBJECTIVETo construct microRNA-223 overexpression and suppression lentivirus vectors and determine their effects after infecting oral squamous cell carcinoma (OSCC) cell line.

METHODSLentivirus vectors GV229 and GV232 were cut by the restriction sites of Age I and EcoR I and connected to the target gene, which contained mature microRNA-223 and microRNA-223 oligonucleotide. Real-time polymerase chain reaction (PCR) method was used to detect the microRNA-223 expression level after infecting the recombinant lentivirus vector into the OSCC cell line.

RESULTSThe successful construction of microRNA-223 recombinant lentivirus vectors was confirmed by the PCR method and DNA sequencing. HN-30 cell infected with microRNA-223 overexpression vector showed a significant increased in microRNA-223 expression, whereas HN-30 cell infected with microRNA-223 inhibitor vector suppressed microRNA-223 expression.

CONCLUSIONThe microRNA-223 overexpression and suppression lentivirus vectors are successfully constructed. These vectors could alter the expression level of microRNA-223 in OSCC cell line significantly, and provide a stable cell line for functional studies in the future.

Objective To investigate the relationship between vascular endothelial function and insulin resistance(IR) and the incidence of metabolic syndrome(MS).Methods A seven-year follow-up study was conducted in 583 simple obese subjects who were chosen in 2000.Body fat,blood pressure,lipid profile,plasma glucose,insulin resistance index(HOMA-IR) and endothelium- dependent dilatation(EDD) were measured before and after the follow-up.Results A total of 177 new cases with MS were found during this seven years,which was 33.78% of all subjects who were followed up in 2007.Cumulative incidence of MS among seven-year was 38.14% in the impaired EDD group and 48.11% in the impaired EDD with IR group,both rates were higher than that of the IR group(25.00%)(P

Objective To synthesize glycylproline p-nitroanilide tosylate as a substrate for glycylprolyl dipeptidyl-aminopeptidase(GPDA),and to study its clinical application.Methods The title compound was prepared by DCC-HOBt synthesis.GPDA in human serum was detected by continuous monitoring method using substrate. Results The substrate with purity of 98.5% was obtained.The linearity of the method was up to 358.1 U/L.Intraassay CV and interassay CV of GPDA in diverse serum samples were 3.01% and 5.04%,respectively.It was shown that GPDA level in patients with hepatic cancer was significantly increased,while those in patients with gastric carcinoma and chronic gastritis were significantly decreased compared with that in the control group in clinical detecting(P

Objective To explore the effects of AP on pulmonary fibrosis. Methods Alveolar macrophages (AM) were treated by AP for 24 hours. Pulmonary fibroblasts (FB) were cultured with the supernatant of AM medium. The protein of transforming growth factor beta 1 (TGF-?1) of AM, the proliferative activity and hydroxyproline (Hyp) content of FB were examined. Rats were treated with AP by intratracheal instillation and sacrificed at 3 days. The TGF-?1 mRNA content in the lung was examined. Results The positive staining macrophages in low and high AP groups and the quantity of TGF-?1 in high AP group were obviously higher than those in the control group (P

Objective To observe the protective effect of asiatic acid on hyperlipidemia golden hamsters induced by high fat diet and explore its mechanism.Methods Hyperlipidemic golden hamsters fed with high-fat diet were administered orally with asiatic acid (8,16,32 mg/kg)for 4 weeks.Levels of serum lipid content,liver histology,hepatic GSH-PX and SOD levels,serum ALT and AST activities were evaluated in golden hamsters,fluorescence quantitative polymerase chain reaction(RT-PCR)is using to examine the liver lecithin cholesterol fatty acyl transferase(LCAT) and class B typeⅠscavenger receptor(SR-BⅠ)mRNA expression. Results Compared with model group,the levels of TC,TG,LDL-C,TC/HDL,ALT and AST in low and mediate asiatic acid groups were all significantly decreased but the levels of SOD and GSH-PX were significantly increased(P<0.01).HE staining results showed that fat deposition in the liver were improved by administration of asiatic acid,and the expression of LCAT and SR-BⅠmRNA were increased.Conclusion Asiatic acid can effectively reduce blood lipid levels,and alleviate liver damage of the hyperlipidemia golden hamsters by lipid regulation and antioxidative ability augmentation.The increased levels of LCAT and SR-BⅠmRNA expression maybe involved in the mechanism of hypolipidaemic effect of asiatic acid.

Objective To explore the imaging findings of branch duct intraductal papillary mucinous neoplasms (BD-IPMNs). Methods A total of 45 BD-IPMNs were confirmed by pathology after surgery.The clinical data and imaging findings were retrospec-tively reviewed.Image analysis included characteristics and the nature of the lesions.Finally,an ROC curve was performed to evaluate the differential diagnostic efficacy between benign and malignant lesions using the imaging findings.Results There were statistically significant differences for differential diagnosis of malignant and benign BD-IPMNs in the maximum diameter of the lesions and the MPD,walls,wall nodules,shape,and the atrophy of pancreatic parenchyma (P < 0.05 ).The diagnostic sensitivity,specificity, AUC and Kappa value of benign and malignant BD-IPMNs were 94.3%,90%,92.1% and 0.81 (P =0.000 1)respectively.Conclu-sion The imaging technology is valuable in diagnosing and differentiating malignant from benign BD-IPMNs.