Characters of one Candida intermedia yeast strain which isolated from nature can produce ethanol from xylose-fermenting been systemic studied. In conditions 28?C, 120 r/min, 72 h, it can produce 6.480 g/L ethanol from 7% xylose and 43.70% theoretical production of ethanol from 3% xylose. It can produce up to 21.225 g/L ethanol when incubation time prolong to 156 h from 8% xylose. It also can ferment 13% glucose produce 47.647 g/L ethanol and reach 76.90% of theoretical ethanol production, respectively. Compared to CK, ethanol productivity can be improved 9.91% when add 8% xylose in three times as 3%, 2% and 3%, respectively. Glucose can be first utilized in the mixture sugar medium. When the ratio of xylose vs. glucose is 3:1in mixture sugar, the productivity of ethanol can be improving 25%.

We developed an R function named "microarray outlier filter" (MOF) to assist in the identification of failed arrays. In sorting a group of similar arrays by the likelihood of failure, two statistical indices were employed: the correlation coefficient and the percentage of outlier spots. MOF can be used to monitor the quality of microarray data for both trouble shooting, and to eliminate bad datasets from downstream analysis. The function is freely avaliable at http://www.wriwindber.org/applications/mof/.
Algorithms ; Computational Biology ; methods ; Internet ; Oligonucleotide Array Sequence Analysis ; methods ; statistics & numerical data ; Reproducibility of Results

We developed an R function named "microarray outlier filter" (MOF) to assist in the identification of failed arrays. In sorting a group of similar arrays by the likelihood of failure, two statistical indices were employed: the correlation coefficient and the percentage of outlier spots. MOF can be used to monitor the quality of microarray data for both trouble shooting, and to eliminate bad datasets from downstream analysis. The function is freely avaliable at http://www.wriwindber.org/applications/mof/.

OBJECTIVETo study the effects of Qingxin Kaiqiao Recipe (QKR) saponin on the expressions of Bax, Bcl-2, beta-amyloid (Abeta), and beta-amyloid precursor protein (betaAPP) in the cortex and hippocampus of Alzheimer's disease (AD) rats.

METHODSThirty-two SD rats of SPF grade were selected. Abeta 25 - 35 was injected into the bilateral amygdala to prepare the AD model. After modeling rats were randomly divided into the model group, the donepezil hydrochloride group (Donepezil Hydrochloride Tablet, 1.67 mg/kg), the QLR group (QLR Decoction, 12.67 mL/kg), and the saponin group (saponin, 6.30 mg/kg), 8 rats in each group. Another 8 rats were selected as the normal group. Rats in the normal group and the model group were given with equal volume of double distilled water by gastrogavage. The intervention was performed once daily for 2 successive weeks. The Morris water maze test was carried out by the end of medication. The escape latency and the platform crossing times were recorded during the 1 -5 days. The expressions of Bax, Bcl-2, Abeta, and betaAPP in the cortex and hippocampus were detected using immunohistochemical assay.

RESULTSCompared with the model group, the 3rd - 5th day escape latency were all shortened in each medication group. The expressions of Bax, Abeta, and betaAPP decreased in the cortex and hippocampus. The numbers of platform crossing increased. The expression of Bcl-2 in the cortex increased. The expression of Bcl-2 in the hippocampus increased in the donepezil hydrochloride group and the QLR group with statistical difference (P<0.01, P<0.05). Compared with the donepezil hydrochloride group, the expression of betaAPP increased in the cortex and hippocampus of the saponin group. The expression of Abeta in the cortex and hippocampus decreased, the expression of Bax in the hippocampus decreased, and the expression of Bcl-2 in the cortex increased in the QLR group. The escape latency was obviously postponed at day 3 -5, the platform crossing times decreased, the expression of Bcl-2 in the cortex and hippocampus decreased in the saponin group, showing statistical difference (P<0.05, P<0.01).

CONCLUSIONQKR could significantly improve AD rats' learning, memory, and spatial capabilities, which might be achieved through decreasing the expressions of Bax, Abeta, and betaAPP in the cerebral cortex and hippocampus, and elevating the expression of Bcl-2.

Alzheimer Disease ; drug therapy ; metabolism ; Amyloid beta-Peptides ; metabolism ; Amyloid beta-Protein Precursor ; metabolism ; Animals ; Cerebral Cortex ; drug effects ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Hippocampus ; drug effects ; metabolism ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology ; bcl-2-Associated X Protein ; metabolism

Objective To construct the Neisseria surface protein A (NspA) DNA vaccine of Neisseria gonorrhoeae and evaluate the humoral and cellular immune responses induced by this vaccine in mice model.Methods The recombinant expression vector pcDNA3.1 (+)/NspA was constructed by inserting NspA gene into the eukaryotic expression vector pcDNA3.1 (+) and confirmed by poly merase chain reaction (PCR),restriction enzymes HindⅢ,XbaⅠand DNA sequencing.NspA mR- NA in transfected RAW264.7 cells and NspA protein expression in transfected COS-7 cells were de- tected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical stai- ning,respectively.Forty-five male BALB/c mice were immunized with pcDNA3.1 (+)/NspA recom binant plasmid.The level of serum anti-Neisseria gonorrhoeae antibody of the immunized mice was detected by tube agglutination test,and the level of interieron (IFN)-?was assayed by enzyme-linked immunosorbent assay (ELISA).The proliferation of splenocytes was determined by methyl thiazolyl tetrazolium (MTT) colormetry.The NspA gene in BALB/c mice was identified by PCR with the total DNA extracted from quadriceps femoris in immunized sites.Results Restriction enzymes digestion a- nalysis and DNA sequencing results revealed that the pcDNA3.1 (+)/NspA had been constructed successfully.NspA gene had been transcripted and expressed in mammalian cells.The peak titer of specific antibody was 1:640 in pcDNA3.1(+)/NspA immunized group and there was no specific an- tibody detected in both pcDNA3.1 (+) immunized group and PBS group.The IFN-?level in pcD NA3.1 (+) immunized group was (23.79?11.85)pg/mL and that in pcDNA3.1 (+)/NspA immu- nized group was(169.71?30.52)pg/mL (P

BackgroundThe quest to look for seed cells is a hot spot of cornea transplant research in solving the problem of the lack of donor. Bone marrow mesenchymal stem cells(BMSCs) have been successfully induced into retinal ganglion cells(RGCs) in vivo,but the successful induction of BMSCs into corneal endothelial cells has not been reported.Objective This experiment was to study the transplantation of BMSCs on corneal endothelial surface using the splitting Descemet's membrane. MethodsFour healthy adult rhesus monkeys were divided into the experimental group ( 3 monkeys) and control group ( 1 monkey). Mesenchymal stem cells (MSCs) were isolated from bone marrow by density gradient centrifugation combined with adhering means. The cultured cells were identified by flow cytometry and its ability to differentiate was determined by allowing them to differentiate into adipocytes in vitro and labeled by 5-bromodeoxyuridine ( BrdU ) for subsequent identification. Corneal grafts of 7 mm in size with tearing of the Descemet' s membrane were prepared in the experimental group and control group. After labeling by 5-bromodeoxyuridine( BrdU ) ,cultured cells were transplanted onto the endothelial surface of cornea grafts in the experimental group, but no cultured cells were seeded in the graft of the control group. The corneal grafts were then sutured in situ, and were removed 1,2 or 3 months after operation to examine the distribution and connection between transplanted cells and their morphologic changes under the electron microscope. Results High purity MSCs were harvested by density gradient centrifugation combined with adhering method. Cultured cells reached confluency after 12 to 16 days, presenting with a spindle shape and parallel or swirling arrangement. Flow cytometry analysis showed that 94.26％ of cells were positive for CD29,7. 51％ for CD34 and 4. 02％ for CD45. Larger nuclei filled with plastosomes, golgiosomes and rough endoplasmic reticula were found on the graft under the transmission electron microscope( TEM ). After 3 weeks, MSCs were differentiated into adipocytes where Oil Red O staining resulted in an orange-red staining in the cytoplasm and blue staining in the nuclei. The transplanted cells attached loosely on the endothelial surface of the corneal graft and came in contact with each other in one month. The shape of the cells appeared as spindle-shaped and polygonal after 2 months and became tightly packed after 3 months. The positive cells retained the BrdU label and presented with brown nuclei. No endothelia cells grew in the cornea graft in the control group, with an absence of BrdU labeling. Conclusions Mesenchymal stem cells can be transplanted onto the corneal endothelial surface successfully and form a monolayer using the centrifugation method, and present with good survival and proliferation ability.

Design space approach is applied in this study to enhance the robustness of first ethanol precipitation process of Codonopsis Radix (Dangshen) by optimizing parameters. Total flavonoid recovery, dry matter removal, and pigment removal were defined as the process critical quality attributes (CQAs). Plackett-Burman designed experiments were carried out to find the critical process parameters (CPPs). Dry matter content of concentrated extract (DMCE), mass ratio of ethanol to concentrated extract (E/C ratio) and concentration of ethanol (CEA) were identified as the CPPs. Box-Behnken designed experiments were performed to establish the quantitative models between CPPs and CQAs. Probability based design space was obtained and verified using Monte-Carlo simulation method. According to the verification results, the robustness of first ethanol precipitation process of Dangshen can be guaranteed by operating within the design space parameters. Recommended normal operation space are as follows: dry matter content of concentrated extract of 45.0% - 48.0%, E/C ratio of 2.48-2.80 g x g(-1), and the concentration of ethanol of 92.0% - 92.7%.
Chemical Precipitation ; Chemistry, Pharmaceutical ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification

OBJECTIVETo study the chemical constituents of an active fraction (DSS-A-N-30) from Danggui Shaoyao San.

METHODDSS-A-N30 was prepared by macroporous resin chromatography, the compound was isolated by column chromatography on silica gel and RPC-18, the structure was elucidated by spectroscopic methods.

RESULTA new monoterpene glycoside was isolated and identified from DSS-A-N-30.

CONCLUSIONThe new monoterpene glycoside was identified as 4"-hydroxyl-albiflorin.

Bridged-Ring Compounds ; analysis ; isolation & purification ; Chromatography, Gel ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Monoterpenes ; analysis ; isolation & purification

OBJECTIVETo study the correlation between the expression of Egr-1 and NF-kappaB and the up-regulation of TNF-alpha and TGF-beta1 in macrophages after stimulation by silica in-vitro.

METHODSMacrophages were treated with antibodies against Egr-1 and NF-kappaB and antisense oligonucleotides. The level of TNF-alpha protein in the cell supernatant was then measured using enzyme-linked immunoadsorbent assay (ELISA). The expression of TGF-beta1 protein was detected by immunocytochemistry. The expression of TNF-alpha and TGF-beta1 mRNAs was also monitored by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSCompared with silica-stimulated macrophages untreated with antibodies, the cells treated with 10 micro g/ml of Egr-1 or NF-kappaB antibodies were associated with reduced expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05). Compared with silica-stimulated untransfected group, the antisense group was associated with obvious reduction in the expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05).

CONCLUSIONThe expression of TNF-alpha and TGF-beta1 mRNAs and proteins are associated with activation of Egr-1 and NF-kappaB in macrophages, after stimulation by silica. It is possible that the corresponding antibodies and antisense oligonucleotides may become a potential therapeutic tool in the management of silicosis in the future.

Animals ; Antibodies ; immunology ; Cells, Cultured ; DNA-Binding Proteins ; genetics ; immunology ; Early Growth Response Protein 1 ; Immediate-Early Proteins ; genetics ; immunology ; Macrophages ; cytology ; metabolism ; Mice ; NF-kappa B ; genetics ; immunology ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Silicon Dioxide ; pharmacology ; Silicosis ; etiology ; Transcription Factors ; genetics ; immunology ; Transforming Growth Factor beta ; biosynthesis ; genetics ; Transforming Growth Factor beta1 ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVETo analyze the outcomes of simultaneous liver resection for patients who have primary colorectal cancer with synchronous hepatic metastases to see if there is any advantage for doing so.

METHODSWe retrospectively analyzed the medical records (1999 - 2009) of 53 consecutive patients with synchronously recognized primary colorectal carcinoma and hepatic metastases who underwent simultaneous (40 patients) or two-stage (13 patients) colonic and hepatic resections performed at our hospital.

RESULTSThere was no thirty-day mortality in both groups. The two groups had significant differences in mean operation duration [(212.9 ± 72.3) min vs. (326.5 ± 140.2) min, P = 0.014], mean blood loss [(337.5 ± 298.0) ml vs. (594.6 ± 430.5) ml, P = 0.020], post-operative hospital stay [(16.2 ± 8.1) day vs. (25.8 ± 8.5) day, P = 0.001]. The incidence rates of post-operative complications were 25.0% (10/40) and 53.8% (7/13), respectively, in the two groups (P = 0.053). The 1-, 3-, 5-year survival rates in the simultaneous resection group were 95.0%, 57.0% and 37.4%, respectively, with a median overall survival of 40.0 months and median disease-free survival of 14.0 months. The 1-, 3-, 5-year survival rates in the two-stage resection group were 92.3%, 58.7% and 36.7%, respectively, with a median overall survival of 38.0 months and median disease-free survival of 13.0 months. There were no significant differences between the two groups in respect of their survivals (P > 0.05).

CONCLUSIONSSimultaneous colectomy and hepatectomy are safe and efficient for colorectal cancer patients who have synchronous colorectal liver metastases, with less complications and blood loss, and shorter hospital stay compared with the two-stage resection.

Blood Loss, Surgical ; Colectomy ; methods ; Colonic Neoplasms ; pathology ; surgery ; Disease-Free Survival ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Humans ; Length of Stay ; Liver Neoplasms ; secondary ; surgery ; Male ; Operative Time ; Postoperative Complications ; Rectal Neoplasms ; pathology ; surgery ; Retrospective Studies ; Survival Rate