A novel raw-starch-digesting glucoamylase producer,Rhizopus sp.W-08,was used in a novel fermentation system of solid-state followed by submerged,and high enzyme activity of 72 IU/mL was obtained.In the following simultaneous saccharification and fermentation process, Saccharomyces cerevisiae Z-06 directly converted raw corn flour to ethanol with the concentration of 21 % (V/V) at 30℃ after 48h.The conversion efficiency of raw corn flour to ethanol was 94.5 % of the theoretical ethanol yield.

BACKGROUND:The method in repair of primary lumbar intervertebral infection is different in different positions, mainly containing anterior, posterior and anteroposterior pathways. In posterior pathway, muscle gap approach is recognized by many fel ows. This program has its special advantage compared with conventional posteromedial approach. OBJECTIVE:To evaluate the clinical effects of debridement, bone graft and internal fixation of pedicle screw placement in repair of primary lumbar intervertebral infection through posterior paraspinal muscle approach. METHODS:Clinical data of 13 patients with primary lumbar intervertebral infection were analyzed retrospectively. There were one case of L 2-L 3 , two cases of L 3-L 4 , four cases of L 4-L 5 and six cases of L 5-S 1 . Lumbar pain was obvious in al patients. Nine cases suffered from radioactive lower limb pain. Al patients received debridement, bone graft and internal fixation of pedicle screw placement through paraspinal muscle approach via posteromedial incision. After treatment, clinical effects were evaluated using Visual Analog Scale and lumbar Japanese Orthopaedic Association scores. RESULTS AND CONCLUSION:Al patients were fol owed up for 12-18 months, no recurrence. X-ray review demonstrated bony fusion, so loosening and breakage were not found in al patients. Visual Analog Scale scores revealed that Visual Analog Scale score was 8.15 preoperatively, 2.15 at 1 week postoperatively, 1 at final fol ow-up, showing significant difference (P<0.05). There was significant difference in pain between pre-treatment and post-treatment, and pain apparently relieved after treatment. Lumbar Japanese Orthopaedic Association score showed that effective outcomes were found in al patients after treatment, including nine cases of excel ent effects, three cases of good effects, one case of average effects, with the excel ent and good rate of 92%. Above findings confirmed that one-stage debridement, bone graft and internal fixation of pedicle screw placement through posterior muscle gap approach provides a good repair method for patients with primary lumbar intervertebral infection. It can achieve intervertebral space directly through intervertebral foramen, retain the central spinous process and lamina, reduce the injury to paraspinal muscle, and keep spinal ligament complex. However, strong spine fixation contributes to bone fusion, and keeps the stability of the spine after repair.

Objective To search for the gene encoding specific antigen of Ancylostoma caninum that induces host′s protective immunity. \ Methods\ A lambda ZAPⅡ cDNA library of A.canium was screened with sera from dogs \{immunized\} subcutaneously with hookworm larvae(L3). After sequencing, insert gene (AcAg) from positive clones was \{ligated\} into PUC18 and PET28C. Recombinant pET28C plasmid was induced to expressed protein in the E.coli BL21. The \{characteristic\} of recombinant protein is analyzed by SDS\|PAGE and Western bloting assay. \ Results \ Five positive \{clones\} were obtained, and proved to be the same. The insert gene (AcAg) in pET28C vector expressed a recombinant \{protein\} of about 43 kDa. Using Western bloting assay, this protein was recognized by the sera from dog immunized with \{Ancylostoma\} caninum third\|stage infected larvae and used for screening library. \ Conclusion \ The AcAg, which \{exhibits\} 35% homologous to Caenorhabditis elegans gene unc\|89, is a novel specific antigen of A.caninum. Its ability to elicit the \{protective\} immunity and the probability of the recombinant protein as a vaccine need to be further evaluated.

Objective:To investigate the regulatory effects of acupuncture and moxibustion on chromatin remodeling complex core catalytic subunit of Brahma-related gene 1(Brg1),histone deacetylase(HDAC)3,HDAC9,and males absent on the first(MOF)in the colon tissue of rats with Crohn disease(CD). Methods:Using the random number table method,60 male Sprague-Dawley rats were divided into 5 groups,including a normal group,a model group,an acupuncture group,a medicinal cake-insulated moxibustion group,and an acupuncture-moxibustion group,with 12 rats in each group.CD rat models were prepared using 2,4,6-trinitrobenzene sulfonic acid(TNBS)in all groups except the normal group.The normal and model groups received no interventions.In the acupuncture group,rats were intervened with acupuncture at bilateral Zusanli(ST36)and Shangjuxu(ST37),20 min/session,once a day.The medicinal cake-insulated moxibustion group received medicinal cake-insulated moxibustion at Qihai(CV6)and bilateral Tianshu(ST25)with 2 cones per point per session,once a day.The acupuncture-moxibustion group received both acupuncture and moxibustion interventions simultaneously.Each intervention was performed for 10 consecutive days.Observations included general condition,disease activity,macroscopic damage,and pathological changes in the rat's colon tissue.Real-time fluorescence quantitative polymerase chain reaction was used to measure the mRNA expression of chromatin remodeling-related enzymes Brg1,HDAC3,and HDAC9,while Western blotting detected the protein expression of Brg1,HDAC3,HDAC9,and MOF in rat's colon tissue. Results:The model group showed significantly increased diarrhea score,occult blood score,macroscopic damage score of colon tissue,and colon macroscopic damage index(CMDI)score,as well as elevated mRNA expression levels of HDAC3 and HDAC9,protein expression levels of HDAC3,HDAC9,and MOF,and decreased mRNA and protein expression levels of Brg1 compared to the normal group(P<0.01).In contrast,compared to the model group,the diarrhea score,occult blood score,macroscopic damage score,CMDI score,mRNA expression levels of HDAC3 and HDAC9,and protein expression levels of HDAC3,HDAC9,and MOF were significantly reduced(P<0.05 or P<0.01),while the mRNA and protein expression levels of Brg1 were significantly increased(P<0.01)in the acupuncture group,the medicinal cake-insulated moxibustion group,and the acupuncture-moxibustion group. Conclusion:Both medicinal cake-insulated moxibustion and acupuncture,either used alone or in combination,can regulate the abnormal expression of chromatin remodeling-related enzymes Brg1,HDAC3,HDAC9,and MOF in the colon tissue,thus reducing colon inflammation in CD rats.

Objective To explore the clinical selection of surgical methods for pure stenosis at the end of the common bile duct.Methods The clinical data of 43 cases of pure stenosis at the end of the common bile duct treated in this hospital from October 2003 to June 2009 were retrospectively analyzed.Of the 43 cases, 12 underwent endoscopic sphincterotomy(EST), 9 sphincteroplasty of Oddi, 5 pancreaticoduodenectomy,8 Roux-en-y choledochojejunostomy and 5 choledocoduodenostomy,4 underwent drainage of the T tube after exploration of the biliary passage.Results Among 37 patients did not undergo the radical pancreaticoduodenectomy,6 had space-occupying lesions 5-19 months postoperatively.Five of them received pancreaticoduodenectomy again.Conclusion Pure stenosis at the end of the common bile duct should be carefully managed.

To investigate the impact of respiratory movement to determine the target volume on cone beam CT (CBCT) for lung tumor, we used CIRS dynamic thorax phantom (Model-CIRS008) to simulate the sinusoidal motion of lung tumor. With a constant amplitude, the ratio of the time of near-end-expiratory and near-end-inspiratory (E/I) changed when it was scanned with CBCT. We analyzed the contrast changes of target by extracting the CT value of each pixel on the center line of the target movement direction. The targets were contoured with region growing method and compared with the motion volume generated by the tumor trajectory method. The result showed that the contrast of near-end-expiratory increased and the contrast of near-end-inspiratory decreased with increasing E/I. The contoured volume generated by region growing method decreased with increasing E/I. When E/I = 4, the amplitude A = 1 cm, diameter of 1 cm and 3 cm target volumes were reduced by 48.2% and 22.7%. The study showed that CBCT was not suitable to be used to accurately determine the range of lung tumor movement. The internal target volume (ITV) may be underestimated in CBCT images.
Cone-Beam Computed Tomography ; Humans ; Lung Neoplasms ; diagnostic imaging ; Movement ; Phantoms, Imaging ; Respiration

Objective To evaluate the characteristics and clinical suitability of the ArcCHECK diode array for volumetric-modulated arc therapy (VMAT) verification.Methods The intrinsic sensitivity,short and long term reproducibility,dose and dose rate dependence,dose per pulsed dependence,field sizes dependence and directional response of the diodes were measured.The results of the diodes were compared with the measurement results of an ionization chamber and calculated results of treatment planning system.Gamma index was used to analyze the dose difference between the calculation and measurement for random selected 211 verified VMAT plans.Results The ArcCHECK performed well for all tests except directional dependence,which varies from a minimum of-3.9％ (seen only when the beam was incident on the diode at 180°) to a maximum of 7.7％ (approximately at 255°).Average gamma analysis passing rates with 3 mm/3％ for 113 nasopharyngeal cancer,48 cervical cancer and 50 rectal cancer patients VMAT plans were 93.5％,95.7％ and 97.5％,respectively,statistical significance has been found between either two of the groups (t=-12.69-4.88,all P＜ 0.01).Conclusions With proposed calibration method,the ArcCHECK diode array is very suitable for VMAT pretreatment verification,the complexity of VMAT plan is the main factor affecting the passing rate.

【Objective】 To study the differentiate expression genes and regulation signaling pathways related to the occurrence and development of dilated cardiomyopathy with heart failure, we used bioinformatics methods to explore gene chip in gene expression omnibus(GEO) and genes related to dilated cardiomyopathy in online mendelian inheritance in man(OMIM) . 【Methods】 GSE21610 and GSE29919 chip gene sets sequenced by Herz- &Diabeteszentrum NRW laboratory of the University of Bochum, Germany with myocardial biopsy specimens from clinical cases under the platform GPL570 and uploaded to GEO public database were selected to perform our study. Myocardial biopsy specimens from dilated cardiomyopathy with heart failure and normal cardiac function were regarded as the experimental group(14 and 21 cases, respectively) and the control group(12 and 8 cases, respectively). Up-regulated expression genes with the criteria: P < 0.05, were screened in GEO2R with the selected samples of two chips, which were then used to perform kyoto encyclopedia of genes and genomes(KEGG) pathways analysis, gene ontology(GO) function analysis, and protein-protein interaction(PPI) analysis, and the results were displayed through Volcano map, Venn map, Heat map, and Bubble charts with enrichment pathways drawn by R language packages. Meanwhile, KEGG pathways with the criteria: NOM. P < 0.05, and core genes relating to dilated cardiomyopathy with heart failure were performed through gene set enrichment analysis (GSEA). All candidate genes were then intersected with the reported genes in the OMIM, respectively, and clinical significance of these candidate genes was explored in relevant literatures. 【Results】 A total of 173 up-regulated expression genes with P < 0.05, were obtained by GEO2R, which are mainly related to inflammatory signals, cell proliferation and differentiation regulating, and classical apoptotic signaling pathways. These genes were intersected with the 754 reported genes in OMIM, and three reported up-regulated expression genes were obtained: NPPA for diagnosis of heart failure, APOA1 that associates with cytokine action, and COL6A1 that regulates lateral tubular remodeling. 158 and 46 core genes from KEGG pathways with NOM. P < 0.05 were obtained by GSEA, respectively, which were then intersected with the 754 reported genes in OMIM, and two core genes were obtained: PRKCA, enhancing myocardial contractility, and BMP2, promoting the development of heart failure. 【Conclusions】 Based on bioinformatics analysis, we found that the candidate genes PRKCA, BMP2, NPPA, and COL6A1 are likely to be closely related to the occurrence and development of dilated cardiomyopathy with heart failure, which can be used to reveal meaningful therapeutic clues and directions for the clinical treatment of dilated cardiomyopathy with advanced cardiac dysfunction.

Objective To determine the expression of miR-486-3p in psoriatic lesions and healthy human skin and to estimate its effect on keratin 17 (K17) expression in HaCaT human keratinocytes.Methods Bioinformatics was used to predict microRNAs that may affect the expression of K17.Tissue samples were obtained from the lesions of 10 patients with psoriasis and normal skin of 10 healthy human controls.RNA was extracted from these tissue samples and reversely transcribed into cDNA with the addition of a Poly (A) tail.Then,real time quantitative PCR was performed to measure the expression of miR-486-3p.Cultured keratinocytes were transfected with miR-486-3p mimics or negative control,and Western blot was performed to determine K17 expression at 48 hours after the transfection.To evaluate the inhibitory effect of miR-486-3p on K17 expression,cultured 293T cells were transfected with the plasmid containing K17 3' untranslated region (UTR) seed sequence,three plasmids containing the complete deletion,interval mutation or double repeats of the seed sequence,or negative control plasmid.At 24 hours after the transfection,a dualluciferase reporter (DLR) assay was performed to quantify the expression of K17.Results Real time PCR showed that the expression level of miR-486-3p was significantly lower in psoriatic lesions than in the normal skin (0.211 ± 0.120vs.0.555 ± 0.425,t =2.62,v =9,P ＜ 0.05).The HaCaT cells transfected with the mimics of miR-486-3p exhibited decreased expression of K17 compared with those transfected with the negative control.DLR assay revealed that the expression level (fluorescence intensity) of K17 in the negative control group was significantly higher than that in the 293T cells transfected with the seed sequence and those with the double repeats of the seed sequence (100.00％ vs.65.31％ ± 6.32％ and 54.18％ ± 10.01％ respectively,both P ＜ 0.05),but did not differ from that in the 293T cells transfected with the complete deletion and interval mutation of the seed sequence (100.00％ vs.114.77％ ± 16.14％ and 110.21％ ± 12.99％ respectively,both P ＞ 0.05).Conclusions The expression of miR-486-3p,which may inhibit K17 expression by binding to the seed sequence of K17 3'UTR,is lower in psoriatic lesions than in normal skin.The decreased expression and inhibitory effect of miR-486-3p may be implicated in the initiation and progression of psoriasis.

0.05).The expression of NF-?Bp65 protein in epithelial ovarian cancer was significantly correlated with FIGO stage(P0.05).There was obviously negative correlation between KISS- 1 and MMP-9 expression in ovarian cancer(rs=-0.547,P