The microbial populations and community structure characterization technologies of the enhanced biological phosphate removal system were reviewed comprehensively in this paper, and their future research directions were outlined.

Objective:In the present study,Bac-to-Bac baculovirus expression system was used to obtain recombinant human Cosmc extracellular domain protein,which can lay the foundation for the research about the structure and function of Cosmc protein in vitro,and simultaneously provide ideas for the research of O-glycosylation and related diseases. Methods: The Cosmc extracellular domain ( Cosmc-ED) gene was cloned into a transfer vector pFastBac1 to form the recombinant donor plasmid pFastBac1-Cosmc ED, which was transformed into competent cells DH10Bac. By using blue-white selection and PCR analysis,we could obtain recombinant shuttle vector rBacmid-Cosmc ED. Then, the recombinant gene DNA of rBacmid-Cosmc ED was used to transfect Sf-9 mediated by cationic lipid formulation,and the recombinant baculovirus bacmid was obtained,which was further used to infect the serum-free cell Sf-9 to express Cosmc-ED in the supernatant. Then the protein of interest was detected by SDS-PAGE and Western blot and purified with Ni-NTA affinity column. Results:SDS-PAGE and Western blot analysis showed a specific band about 33 kD,consistent with the interest protein. Mass spectrometry results further prove that the protein was Cosmc extracellular domain protein. Conclusion: Human Cosmc-ED protein can be successfully expressed in Sf-9 insect cells and laid basis for subsequent studies.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; pathology ; radiotherapy ; Female ; Humans ; Lung Neoplasms ; pathology ; radiotherapy ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Radiotherapy, Conformal ; methods ; Remission Induction

Objective To investigate the effects of sex hormone-binding globulin (SHBG)gene in the apoptosis of human trophoblastic cells.Methods The siRNA specific-targeting SHBG gene was transfected into human trophoblastic cells and they were divided into six groups:trophoblasts without transfection in normal control groups(group Ⅰ);transfect liposome in blank control groups(group Ⅱ);transfect nonspecific siRNA in negative control groups(group Ⅲ);transfect SHBG siRNA-Ⅰ,SHBG siRNA-Ⅱ,SHBG siRNA-Ⅲ respectively in trans-fection group(group Ⅳ,Ⅴ,Ⅵ).Hoechst 33258 dying method was used to detect cell apoptosis.SHBG and Caspase-3 mRNA profiling and the level of SHBG and caspase-3 protein were detected by real-time PCR and Western blot.Results There was no statistical significant difference in the gene expression and protein level of SHBG and caspase-3 in group Ⅰ,Ⅱ and Ⅲ (P >0.05).In Ⅳ,Ⅴ and Ⅵ group,there was no statistical significant difference in the expression level of SHBG and caspase 3 (P >0.05).Compared with group Ⅰ,Ⅱ and Ⅲ,the a-mount of SHBG gene expression decreased obviously,the caspase-3 mRNA and protein level increased obviously and the trophoblast cell ap-optosis increased markedly (P <0.05).Conclusion Through siRNA interference technology can reduce SHBG gene expression in human trophoblastic cells,and it can lead to excessive apoptosis of human trophoblasts cells.

Although the essential oil of Xiangfu Siwu decoction (XFSWD) has strong pharmacological activity, its special physical and chemical properties restrict the clinical application and curative effect. In this paper, Xiangfu Siwu decoction essential oil (XFS-WO) was prepared by forming inclusion complex with β-cyclodextrin (β-CD). The present study is to investigate the effect of β-CD inclusion complex on the transport of major components of XFSWO using Caco-2 cell monolayer model, thus to research the effect of this formation on the absorption of drugs with low solubility and high permeability, which belong to class 2 in biopharmaceutics classification system. A sensitive and rapid UPLC-MS/MS method was developed for simultaneous quantification of senkyunolide A, 3-n-butylphthalide, Z-ligustilide, dehydrocostus lactone and α-cyperone, which are active compounds in XFSWO. The transport parameters were analyzed and compared in free oil and its β-CD inclusion complex. The result revealed that the formation of XFSWO/β-CD inclusion complex has significantly increased the transportation and absorption of major active ingredients than free oil. Accordingly, it can be speculated that cyclodextrin inclusion complex can improve bioavailability of poorly water-soluble drugs. Above all these mentioned researches, it provided foundation and basis for physiological disposition and pharmaceutical study of XFSWD.
Biological Transport ; Caco-2 Cells ; Drugs, Chinese Herbal ; analysis ; Humans ; Oils, Volatile ; analysis ; beta-Cyclodextrins ; pharmacology

Objective To evaluate enteral nutrition treatment on nutritional status of patients with esophageal cancer during radiotherapy in Chinese population. Methods We searched domestic and foreign relevant databases (PubMed, Medline, Cochrane Library, Wanfang Data, Chinese Journal Database, VMIS) by key words (esophageal cancer, chemoradiotherapy, enteral nutrition) , meeting the conditions of the clinical randomized controlled trials, and evaluated the quality of each document. We used meta-analysis method to analysis the published literatures about clinical randomized controlled trials, which were enteral nutrition treatment on nutritional status of patients with esophageal cancer during radiotherapy in Chinese population.RevMan5.1 statistical analysis software were used. Results Seven separate clinical randomized controlled trials including 503 cases were included into the meta-analysis. The following five indexes of nutritional status in enteral nutrition treatment (ENT) group were superior to routine treatment (RT) group (P<0.05): body weight, Body Mass Index (BMI), hemoglobin, serum albumin, serum prealbumin. The MD Values and 95％CI (confidence interval) were 5.54 (3.61-7.46), 2.20 (2.02-2.39), 12.46 (5.86-19.05), 4.04 (2.48-5.61), 49.53 (20.93-78.13) .The funnel plot showed no significant publication bias in each indexes. Conclusion Enteral nutrition treatment can improve the nutritional status of patients with esophageal cancer during radiotherapy compare only with routine treatment.

Objective To explore the effect of calcium dobesilate on the apoptosis of retinal cells in diabetic rats .Methods Thirty-six Wistar rats were randomly into three groups:normal group ( n =12 ) , model group(n=12), test group(n=12).The rat in model and test groups of diabetes was given with high fat diet for 8 weeks, and then was made by intraperitoneal injection of streptozotocin ( STZ ) .After successfully es-tablished the diatetes animal model , the rats in test group was lavaged with 150 mg? kg -1? d-1 calcium dobesilate continuous 8 weeks, the rats in normal and model group were given same amount of normal saline . Neuronal apoptosis in retina was detected by TdT -mediated dUTP nick end labeling ( TUNEL ) . The expression of B -cell lymphoma -2 ( Bcl-2 ) , Bcl -2 assaciated X protein ( Bax ) and p53 protein and mRNA were assayed by Western blot and reverse transcription -polymer-ase chain reaction.The activity of cysteinyl aspartate specific proteinase 3 ( Caspase 3) was measured by spectrophotometry .Results Compared with normal group , apoptotic index of retina , the expression of Bax , p53 protein and mRNA , the activity of Caspase 3 was increased ( P<0.01 ) , the expression of Bcl -2 protein and mRNA was decreased in model group ( P<0.01 ) . The expression of Bax , Bcl -2 and p53 mRNA was ( 0.20 ±0.03 ) vs ( 1.00 ±0.09 ) , ( 1.39 ±0.10 ) vs (0.21 ±0.02);(0.28 ±0.05) vs (0.88 ±0.08), in normal group and model group, respectively.Compared with model group,apoptotic index of retina (12.40 ±1.24), the expression of Bax (0.57 ±0.05) and p53 (0.35 ±0.03) protein, the expression of Bax ( 0.56 ±0.05 ) and p53 ( 0.36 ±0.04 ) mRNA, the activity of Caspase 3 (1.72 ±0.17) in test group were all decreased.While the expression of Bcl -2 protein (0.63 ±0.06) and mRNA (0.74 ±0.04 ) were increased in test group ( all P<0.01 ) .Conclusion The calcium dobesilate inhibit neuronal apoptosis by regulation expression of cell apoptotic related protein .

BACKGROUNDIt has been reported that CD8(+) regulatory cells could be induced upon oral tolerance. The purpose of this study was to investigate the changes of CD8α(+) T cells in dextran sulfate sodium (DSS)-induced colitis mice pretreated by oral immune regulation.

METHODSThe effects of five low oral doses of colitis-extracted proteins (CEP) on colitis were evaluated by clinical manifestation and histological lesions. The percentages of CD8α(+) T cells gating on CD3(+) T cells were evaluated in the gut-associated lymphoid tissues (GALT) and the spleens by flow cytometry. Differences between the two groups were compared by Student's t test or Mann-Whitney U test.

RESULTSCompared to bovine serum albumin (BSA)-fed control mice, administration of CEP resulted in marked alleviation of colitis. The proportion of CD8α(+) T cells, not only in intraepithelial lymphocytes (IELs) and lamina propria lymphocytes (LPLs) of the large intestine (LI) but also in spleen from CEP-fed colitis mice, was significantly higher than that from BSA-fed colitis mice (LI-IELs: (71.5 ± 5.4)% vs. (60.1 ± 4.3)%, P < 0.01; LI-LPLs: (60.7 ± 5.2)% vs. (51.9 ± 4.7)%, P < 0.01; spleen: (24.1 ± 3.6)% vs. (20.3 ± 4.1)%, P < 0.05; n = 8). Mucosal repair in repair-period mice five days after termination of DSS treatment was also accompanied by an increase of CD8α(+) T cells in large intestinal mucosal lymphocytes (LI-IELs: (72.1 ± 3.7)% vs. (61.5 ± 4.5)%, P < 0.01; LI-LPLs: (62.1 ± 5.7)% vs. (52.7 ± 3.6)%, P < 0.01; n = 8). The proportion of CD3(+) T cells increased in Peyer's patches (PPs) and decreased in mesenteric lymph nodes (MLNs) from colitis mice compared to untreated mice, whereas the change pattern of CD3(+) T cells in PPs and MLNs from CEP-fed colitis mice was just on the contrary.

CONCLUSIONImprovement of DSS-induced colitis resulted from oral immune regulation is associated with an increase in CD8α(+) T cells in spleen and large intestinal mucosa.

Administration, Oral ; Animals ; CD8-Positive T-Lymphocytes ; metabolism ; Colitis ; chemically induced ; complications ; Dextran Sulfate ; toxicity ; Flow Cytometry ; Lymphocytes ; cytology ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Proteins ; administration & dosage ; immunology ; Spleen ; cytology ; metabolism

Objective: To investigate hepatitis C virus(HCV)genotyping and the serum HCV-RNA concentration in patients infected with different HCV genotypes and to provide information for evaluation of disease condition and anti-viral treatment efficacy. Methods: A total of 60 anti-HCV positive serum samples were collected before antiviral treatment. RT-PCR was performed for the 5′ non-cording region and was followed by nucleotide sequencing for HCV genotyping. Meanwhile, serum HCV-RNA concentration was detected by quantitative PCR. SPSS21.0 and Graphpad Prism 5.0 software were used for data analysis. Analysis of variance (ANOVA) was used for comparison among multi-groups and the t-test was used for comparison between two groups. Results: The frequencies of HCV genotypes 1b, 3a, 1a and 2a were 48.3% (29/60), 23.3% (14/60), 16.7% (10/60) and 10% (6/60), respectively. And, there is one subtype 2c was detected in this study. The mean serum viral concentration with standard deviation of HCV in genotype 1a, 1b, 2a, and 3 a were 5.46±1.19, 6.22±0.78, 5.47±0.65, and 5.38±0.98 log₁₀ (IU/ml) respectively. Conclusions The infection rate of HCV genotype 1 was significantly higher than that of genotype 2 and 3 (P<0.01). The statistical analysis showed the serum HCV-RNA concentration in patients with subtype 1b was significantly higher than that of the subtype group 1 a, 2 a, and 3 a (P<0.05). The study of the relationship between HCV genotypes and the serum HCV-RNA concentration may contribute to anti-viral treatment prescription for hepatitis C patients.

Major surface protein (p30) and Dense Granule Antigen GRA6 of Toxoplasma gondii have good antigenicity, and could be used for detection of IgM against Toxoplasma gondii. GRA6 may complement P30 to reach more high sensitivity for detection of antibodies to Toxoplasma gondii, so, we try to express the chimeric protein of GRA6 and P30 by genetic engineering, identify its antignenicity and use for developing diagnosis reagent. Antigenic domains of p30 and GRA6 of Toxoplasma gondii were screened by analyzing their sequences using the software ANTHEWIN. Two DNA fragments encoding respectively antigenic domains of p30 and GRA6 were cloned, they were inserted into the same expression vector pET28a( + ) and expressed as a chimeric protein in Escherichia coli. BL21(DE3), the expressed chimeric protein of p30 with GRA6 in a form of inclusion body was about 25% of total proteins of E. coli. BL21(DE3). The inclusion body was washed once with 0.5% Triton X-100 and dissolved with 0.5% SKL, after renaturation by gradient dialysis, the recombinant protein was purified by DEAE-Sepharose FF cation column and then detected with 12% SDS-PAGE, it exists mainly in the eluted peak with 300 mmol/L NaCl and has high purity. By using enzyme-linked immunosorbent assay (ELISA), the recombinant protein was examined for reactivity with immunoglobulin M (IgM) antibodies in 6 sera from patients infected with Toxoplasma gondii ., it was reactive with all the 6 sera but not with sera from normal people, these results showed that the recombinant chimeric antigen has good antigenicity and specificity and could be used for detection of IgM against Toxoplasma gondii. The expressed chimeric protein could be used for epidemic investigation of Toxoplasma gondii, blood donor screening, especially for detection of pregnant women, and is of great significance in prevention of Toxoplasma gondii infection.
Animals ; Antigens, Protozoan ; genetics ; immunology ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin M ; immunology ; Models, Genetic ; Polymerase Chain Reaction ; Pregnancy ; Protozoan Proteins ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Toxoplasma ; genetics ; immunology