This study aims to investigate the virological impact of the stalk region and cysteine (C) in neuraminidase (NA) of influenza A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1) viruses. The NA of A/ Anhui/1/05 (H5N1), defined as AH N1, lacked 20 amino acids (including C, defined as s20) as compared with NA of A/Ohio/07/2009 (H1N1) (defined as 09N1). We deleted s20 of 09N1 to construct 09N1-s20, and inserted s20 into AH N1 to construct AH N1+s20. To investigate the impact of C on the biological function of NA, we deleted C in 09N1 to construct 09N1-C and inserted C into AH N1 to construct AH N1-C. The pseudo-type viral particle (pp) system was used to evaluate the impact of these mutants on virology. The combination of 09N1-C and 09H1 (defined as 09H1::09N1-C) showed an infectivity 8 times that of the wild type 09H1::09N1, while the infectivity of the combination of AH N1+C and AH H5 (defined as AH H5::AH N1+C) was much lower than that of the wild type AH H5::AH N1. The infectivity of the combination of 09N1-s20 and 09H1 (defined as 09H1::09N1-s20) was 4 times that of the wild type 09H1::09N1; the infectivity of the combination of AH N1+s20 and AH H5 (defined as AH H5:: AH N1+s20) was 1/7 that of the wild type AH H5::AH N1. The co-existence of 09N1-C and AH H5 displayed 6 times the infectivity of AH H5::09N1, while the infectivity of 09H1::AH N1+C was very low. Multimer analysis showed that in the wild type 09N1, the forms of NA were dimer > tetramer > monomer; the major component of NA in 09N1-C was monomer; in 09N1-s20, the forms of NA were monomer > dimer. AH N1 was mainly composed of monomer; in AH N1+s20, the forms of NA were dimer > monomer > tetramer; in AH N1+C, the forms of NA were dimer > tetramer. Deletion of C or s20 from 09N1 did not change the expression of NA. The study suggested that deletion of C from the stalk region of NA in A/Ohio/07/2009 (H1N1) increases infectivity. Insertion of C into NA's stalk region of A/ Anhui/1/05 (H5N1) significantly decreases infectivity. Cysteine deletion in the stalk region is important for the infectivity of A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1). It may interfere with the infectivity via changes in NA polymerization.
Amino Acid Motifs ; Humans ; Influenza A Virus, H1N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza A Virus, H5N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza, Human ; virology ; Neuraminidase ; chemistry ; genetics ; metabolism ; Viral Proteins ; chemistry ; genetics ; metabolism ; Virulence

In this study, the general bacterial probe and specific cellulolytic bacterial probes were used to quantify the bacteria in rumen. The total RNA were extracted and then hybridized with general bacterial probe after a dilution of concentration. The result showed that there was a high correlation between the hybridization signal and the dilution of total bacterial RNA. Based on the result above, the quantities of three cellulolytic bacteria in rumen sample were detected. The comparative RNA percentage of three cellulolytic bacteria to total bacterial RNA were similar to the previous reports. It can be concluded that the quantification of bacteria in rumen could be conducted by this approach, and which could be used in future research.

OBJECTIVETo explore the feasibility and safety of abdominosacral resection for patients with locally advanced primary low rectal cancer.

METHODSA total of 97 low rectal cancer patients were amenable to surgery but not anal sphincter preservation were included in this study and divided into the abdominoperineal resection group(n=49) and abdominosacral resection group(n=48) according to the order of alternative admission time between June 2010 and January 2012. Intraoperative and postoperative parameters were compared between the two groups.

RESULTSThe surgery went well and no perioperative mortality in the two groups. Compared with abdominoperineal resection group, the operative time of abdominosacral resection group(including the 2nd position adjustment time) was longer[(188±45) min vs. (143±48) min, P=0.000], the unexpected prostate or vagina injury incidence was lower [0 vs. 14.3%(7/49), P=0.032), and the perineal wound infection rate was lower [2.1% (1/48) vs. 18.4% (9/49), P=0.040].

CONCLUSIONAbdominosacral excision is feasible and safe for patients with locally advanced primary low rectal cancer.

Abdomen ; surgery ; Digestive System Surgical Procedures ; Humans ; Perineum ; Rectal Neoplasms ; surgery ; Treatment Outcome

OBJECTIVETo assess the reaction of cytokines induced killer (CIK) cells treatment in hematopoietic injury at different levels on patients with benzene poisoning and seek a novel, safe and effective immunotherapy for benzene poisoning.

METHODSCIK cells were in vitro activated by interleukin-2 (IL-2) and granulocyte-macrophage-colony-stimulating factor (GM-CSF) from the peripheral blood mononuclear cells (PBMC). Thirty-two patients with benzene poisoning were treated with CIK cells. Nineteen patients with mild or moderate benzene poisoning in the control group were treated with VitB4, batilol, leucogen, inosine and stanozolol. The results for treatment of 12 patients with aplastic anemia induced by severe benzene poisoning (the efficacy rate and the case fatality rate) were analyzed. The change of T-lymphocyte subset analyzed by flow cytometry was also observed before and after treatment.

RESULTSFor mild or moderate benzene poisoning, the increase of WBC and RLT in CIK group was higher than that in the control group (P < 0.05). The CD(4)/CD(8) levels were significantly increased after CIK treatment. And for severe benzene poisoning, the effective rate of the CIK group was 91.7% and the mortality rate was 0%.

CONCLUSIONCIK treatment is safe and effective for hematopoietic injury caused by benzene poisoning. The mechanism may be related with the immune modulation of CIK treatment on immunodeficiency of patients with benzene poisoning.

Adult ; Benzene ; poisoning ; Cytokine-Induced Killer Cells ; immunology ; Female ; Follow-Up Studies ; Humans ; Immunotherapy ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the effects of sub-micro emulsion composition on cellular uptake and disposition of incorporated lipophilic drug.

METHODSSub-micro emulsions containing 10 % oil, 1.2 % lecithin and 2.25 % glycerol were prepared, and the fluorescent agent coumarin 6 was used as a model drug. The effects of oil types, co-surfactants and cationic lipid on uptake and elimination kinetics of 6-coumarin in HeLa cells were studied. The uptake mechanism of sub-micro emulsions was further investigated.

RESULTSOil type and Tweens had no influence on the cellular uptake. Modifications of surfactants with Span series increased the cellular influx, among which Span 20 with hydrophilic-lipophilic balance (HLB) value of 8.6 was the best enhancer. The intracellular drug level reached up to (46.09 ± 1.98)ng/μg protein which had significant difference with control group [(38.54 ± 0.34)ng/μg protein]. The positively charged emulsions significantly increased the uptake rate constant and elimination rate constant which were 4 times and 1.5 times of those in anionic groups, respectively. The uptake enhancement was also observed in cationic emulsions, cellular concentrations at plateau were (42.73 ± 0.84)ng/μg protein, which was about 3 times of that in anionic emulsions [(15.71 ± 0.74)ng/μg protein], when extracellular drug concentration kept at 100 ng/ml. Cationic emulsions delivered the payload mainly by direct drug transfer to contacted cells, while the negative ones depended on both drug passive diffusion and clathrin-mediated endocytosis of drug containing oil droplets which accounted for 20% of the intracellular drug.

CONCLUSIONInterfacial characteristic of sub-micro emulsions such as co-surfactants HLB as well as zeta potentials can influence lipophilic drug both in cellular uptake and elimination.

Anions ; Cations ; Coumarins ; pharmacokinetics ; Emulsions ; Endocytosis ; HeLa Cells ; Humans ; Surface-Active Agents ; pharmacokinetics ; Thiazoles ; pharmacokinetics

A new algorithm of adaptive super-resolution (SR) reconstruction based on the regularization parameter is proposed to reconstruct a high-resolution (HR) image from the low-resolution (LR) image sequence, which takes into full account the inaccurate estimates of motion error, point spread function (PSF) and the additive Gaussian noise in the LR image sequence. We established a novel nonlinear adaptive regularization function and analyzed experimentally its convexity to obtain the adaptive step size. This novel algorithm can effectively improve the spatial resolution of the image and the rate of convergence, which is verified by the experiment on optical images.
Algorithms ; Image Processing, Computer-Assisted ; methods ; Motion ; Time Factors

OBJECTIVETo find the status and changes of the soil nutrients in rhizosphere of Atractylodes lancea.

METHODTotal nitrogen (total N), available K, available P, organic matter (ORG), available nitrogen and pH in rhizosphere soil of the wild growing A. lancea in 3 sites, MS, LT and MFS, and the cultivated ones with different ages in LT were detected.

RESULTThe contents of total nitrogen (total N), available K, available P, organic matter (ORG), available nitrogen and pH value in rhizosphere soil were significant different between MS, LT and MFS (P < 0.01). The results of the 6 detected parameters in MS were the lowest, in MFS were the highest and in LT were in the middle. The total N, ORG and available N in the cultivated A. lancea were lower than that in the wild ones (P < 0.01) and available P and pH value in the cultivated A. lancea were higher than that in wild ones (P < 0.01) and there was no difference in available K between the wild and cultivated ones in LT (P > 0.05); 3 available P in rhizosphere soil of the two years old A. lancea were higher than of the one year old A. lancea (P < 0.01) and there were no difference of total N, ORG, available N, available K and pH value in rhizosphere soil of A. lancea between one year and two years plant (P > 0.05).

CONCLUSIONIt is indicated that the growth of A. lancea in Mt. Mao is faced nutrient stress.

Atractylodes ; growth & development ; China ; Ecosystem ; Hydrogen-Ion Concentration ; Nitrogen ; analysis ; Organic Chemicals ; Phosphorus ; analysis ; Plants, Medicinal ; growth & development ; Potassium ; analysis ; Rhizome ; growth & development ; Soil ; analysis

OBJECTIVETo explore the effects of Huoxiang Zhengqi liquid (HXZQ) on enteric mucosal immune responses in mice with Bacillus dysenteriae and Salmonella typhimurium induced diarrhea (BSD).

METHODMice were randomly divided into four groups with 10 mice in each group: control group (control), BSD group, Huoxiang Zhengqi liquid treated BSD groups at high dosage and low dosage (HXZQ high, HXZQ low). HXZQ was administrated from the day of diarrhea induction at dosage of 5.21 g kg(-1) and 0.52 g kg (-1) respectively. Peyer's patch and periphery lymphocytes were prepared for flow cytometry, and level of TNF-alpha in periphery and enteric tissue homogenate were determined with ELISA. Student's t-test was used for statistics.

RESULTMice in BSD group started showing continuous diarrhea at the day of induction till the fourth day when the mice were sacrificed. Diarrhea in the mice of HXZQ high and low groups lasted for 36 and 54 h respectively. There were more CD4+ and CD8+ cells in periphery, less CD4+ cells in peyer's patch in BSD mice comparing to normal mice. In peyer's patch, there were more CD8+ cells in mice in HXZQ high and low groups and more CD4+ in mice in HXZQ high group. Higher level TNF-alpha in periphery and intestinal tissue homogenate in BSD group were observed. Mice in HXZQ high group showed the decreased level TNF-alpha in periphery and enteric tissue homogenate.

CONCLUSIONThe immune regulation on peyer's patch CD4+ and CD8+ cells and suppression on TNF-alpha level in enteric homogenate might partially explain the effect of HXZQ on improvement of BSD.

Animals ; CD4-CD8 Ratio ; Colon ; immunology ; metabolism ; pathology ; Diarrhea ; immunology ; metabolism ; microbiology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Dysentery, Bacillary ; immunology ; metabolism ; microbiology ; Immunity, Mucosal ; drug effects ; Intestinal Mucosa ; immunology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Peyer's Patches ; drug effects ; immunology ; pathology ; Plants, Medicinal ; chemistry ; Random Allocation ; Salmonella Infections ; immunology ; metabolism ; microbiology ; Salmonella typhimurium ; immunology ; Shigella dysenteriae ; immunology ; T-Lymphocyte Subsets ; drug effects ; immunology ; pathology ; Tumor Necrosis Factor-alpha ; blood ; metabolism

Objective To investigate the biofilm formation rate of Candida albicans strain and its sensitivity to antifungal agents.Methods Candida albicans strains were isolated from 165 patients with vulvovaginal candidiasis (VVC) or recurrent vulvovaginal candidiasis (RVVC) in our hospital.The sensitivity of C.albicans strains to antifungal agents was evaluated by drug sensitivity test.And C54 strains for subsequent experiments in vivo and vitro were selected.Biofilms of single microbe or polymicrobial combination were induced in vitro,and their minimal inhibitory concentration (MIC) values were determined in free and biofilm patterns,respectively.C.albicans and mixed microbe suspensions were injected into mice to establish a model of VVC.Using ELIASA,OD595 values were determined to observe biofilm growth rates across groups.Results The largest number of itraconazole-resistant on C.albicans strains (4.20％).MIC values of all groups were higher in a free pattern than in a biofilm pattern.The polymicrobial biofilms formed by co-culture of fugal and bacterial strains were markedly resistant to various antifungal agents.Mter induction in the biofilm pattern,for C54 + Escherichia coli group,MIC values of resistance to amphotericin B,itraconazole,5-flucytosine and fluconazole were 512,＞256,＞512,and ＞ 1024 μg · mL-1;for C54 + Streptococcus agalactiae group,MIC values of resistance to above antifungal agents were ＞512,＞256,＞512,and ＞ 1024 μg · mL-1.The biofilm formation rate was significantly higher in vaginal samples isolated from the mice who were injected with mixed microbe suspension than those who were injected with standard C.albicans suspension (P ＜ 0.05).For ATCC14053 group,OD595 values were 0.20 ± 0.11,0.24 ± 0.024,0.25 ± 0.06 at 24,48 and 72 h,respectively;for C54 + E.coli group,OD595 values were 0.69 ±0.88,0.79 ±0.65,1.10 ±0.64 respectively;for C54 +S.agalactiae group,OD595 values were 0.68 ±0.42,0.81 ± 0.77,1.10 ± 0.10,respectively.Conclusion Polymicrobial biofilm formation can improve the biofilm formation rate of C.albicans and its resistance to antifungal agents and will enhance vaginal injury in patients with RVVC.

OBJECTIVETo prepare reference samples of Mycobacterium tuberculosis culture filtrate protein-10 (CFP-10) and CFP10-streptavidin fusion proteins (CFP10/SA) for time-resolved fluoroimmunoassay (TRFIA).

METHODSThe CFP10 gene was amplified by PCR from Mycobacterium tuberculosis strain H37Rv and cloned into pET24b, pET24b-streptavidin (SA) or pET21a-SA expression vectors. The recombinant proteins CFP10, CFP10-SA and SA-CFP10 were expressed in Rosetta cells, purified via nickel affinity chromatography and refolded by dialysis. The sensitivity and stability of the resultant proteins as reference samples were evaluated by double-antibody sandwich TRFIA.

RESULTSCFP10-SA and SA-CFP10 fusion proteins were expressed as inclusion bodies, whereas CFP10 was expressed in a soluble form. The resultant purity of the 3 recombinant proteins all exceeded 95%. TRFIA results showed that CFP-SA fusion protein possessed the best sensitivity (0.02 µg/L) and stability.

CONCLUSIONThe reference samples of CFP10 for TRFIA detection have been successfully prepared and can be used in the development of a diagnostic kit for Mycobacterium tuberculosis.

Bacterial Proteins ; genetics ; standards ; Fluoroimmunoassay ; methods ; Gene Amplification ; Mycobacterium tuberculosis ; isolation & purification ; Reference Standards