Soluble cello-oligosaccharide with 2-6 oligosaccharide units is a kind of oligosaccharide with various biological functions, which can promote the proliferation of intestinal probiotics such as Bifidobacteria and Lactobacillus paracei. Therefore, it has a regulatory effect on human intestinal microbiota. In this study, a Cc 01 strain was constructed by expressing cellodextrin phosphorylase (CDP) in Escherichia coli. By combining with a previously constructed COS 01 strain, a three-enzyme cascade reaction system based on strains COS 01 and Cc 01 was developed, which can convert glucose and sucrose into cello-oligosaccharide. After optimization, the final titer of soluble cello-oligosaccharides with 2-6 oligosaccharide units reached 97 g/L, with a purity of about 97%. It contained cellobiose (16.8 wt%), cellotriose (49.8 wt%), cellotetrose (16.4 wt%), cellopentaose (11.5 wt%) and cellohexose (5.5 wt%). When using inulin, xylo-oligosaccharide and fructooligosaccharide as the control substrate, the biomass (OD₆₀₀) of Lactobacillus casei (WSH 004), Lactobacillus paracei (WSH 005) and Lactobacillus acidophilus (WSH 006) on cello-oligosaccharides was about 2 folds higher than that of the control. This study demonstrated the efficient synthesis of cello-oligosaccharides by a three-enzyme cascade reaction and demonstrated that the synthesized cello-oligosaccharides was capable of promoting intestinal microbial proliferation.
Humans ; Oligosaccharides ; Biomass ; Escherichia coli/genetics* ; Gastrointestinal Microbiome ; Glucose

Objective:To determine the seroprevalence of celiac disease in susceptible population, and to analyze the relationship between demographic characteristics, dietary habits, lifestyle and serological positivity so as to provide guidance for the prevention and treatment of celiac disease in Southern China.Methods:A total of 1 273 individuals who participated in Guangdong Province Health Screening Program in 2015, were selected as serologically positive subjects of celiac disease, including people with irritable bowel syndrome, colitis, diarrhea, anemia, low BMI, short stature, type 1 diabetes mellitus (T1DM), rheumatoid arthritis (RA), ankylosing spondylitis, psoriasis and bristol grade=6 or 7. All subjects were tested for serum IgA anti-tissue transglutaminase antibodies (TTGA), IgA antibodies against deamidated gliadin peptides(DGPA) and IgG against deamidated gliadin peptides (DGPG). Dietary habits, lifestyle and demographic characteristics were compared in subgroups.Results:The seroprevalence of celiac disease in susceptible population was 0.94% (95% CI 0.54%-1.64%) including 0.08% (1/1 273) for TTGA, 0.47% (6/1 273) for DGPA, and 0.39% (5/1 273) for DGPG. The seropositive rate was 3.6% (1/28) in patients with psoriasis, 2.1% (2/95) in the low BMI group, 1.9% (1/53) in T1DM group, 1.8% (3/169) in diarrhea group and 1.1% (5/463) in RA group. No significant difference was found in age, gender, high carbohydrate diet or lifestyle between the negative and the positive subjects. Conclusions:In Southern China, the seropositive rate of celiac disease is 0.94% in susceptible population, which prompts an urgent need of serological screening for early diagnosis.

Objective To optimize the detection test of Pig-a gene mutation in peripheral blood of rats by enriching and detecting mutant erythrocytes, using immunomagnetic separation technique in combination with flow cytometry. Methods SD rats were administered with 20,40 and 80 mg/(kg·bw)doses of N-ethyl-N-nitrosourea(ENU) continually for 3 days. The peripheral blood samples of rats were collected on the 7th,14th and 28th days, respectively, after treatment. Immunomagnetic separation columns were used to enrich RETCD59- and RBCCD59- cells, and then flow cytometry was used to count the number of pre-column and post-column peripheral erythrocytes. Results Compared with the control group,the frequencies of RETCD59- and RBCCD59- were significantly increased in each ENU group(P<0.05). With immunomagnetic separation technique, the test of Pig-a gene mutation of a sample could be completed within 3 minutes,and the number of detected RETCD59- or RBCCD59-cells was up to 2×104or 9×104, respectively. Conclusions In this study,immunomagnetic separation in combination with flow cytometry is used to establish and optimize the Pig-a gene mutation test in rat peripheral blood,showing a high-throughput detection and improved accuracy and efficiency of the experiment.

Objective To explore the feasibility of intravoxel incoherent motion DWI (IVIM-DWI) in differential diagnosis of hepatocellular carcinoma (HCC) and focal nodular hyperplasia (FNH).Methods A total of 407 patients with clinically-suspected HCC or FNH underwent conventional and dynamic enhanced MRI and IVIM-DWI,60 patients (40 cases of HCC,20 cases of FNH) were enrolled.Parameters of ADC,slow apparent diffusion coefficient (D),fast apparent diffusion coefficient (D*) and fraction of fast apparent diffusion coefficient (f) were obtained by monoexponential model and biexponential model respectively.Results The values of ADC,D,D* and f in FNH group were (1.60±-0.25) × 10-3mm2/s,(1.12±0.17)×10-3mm2/s,(44.89±18.23)× 10-3 mm2/s and (34.80 ± 9.68)％,and those in HCC group were (1.32 ± 0.21) × 10-3 mm2/s,(0.82±-0.21) × 10-3mm2/s,(49.82±20.11) × 10 3mm2/s and (28.72±13.84) ％,respectively.Significant inter-group differences were observed in ADC and D (both P＜0.001),however,there were no significant differences in D* and f (both P＞0.05).The areas under the ROC curve of D were 0.90,and taking D=0.96 × 10-3 mm2/s as cut-off value,the sensitivity and specificity of D in diagnosis of HCC were 84.44％ and 90.02％.Conclusion IVIM-DWI is useful to distinguish FNH from HCC,and the D value in biexponential model has the best diagnostic efficacy for differentiations.

OBJECTIVEPsoriasis is an autoimmune-related chronic inflammatory skin disease strongly associated with the dysfunction of Th17 cells. Retinoic acid-related orphan nuclear receptor γt (RORγt) plays a critical role in the differentiation and maturation of Th17 cells and in cell-derived immunologic derangement. We conducted this study to investigate potential mechanism by which the derivative of digoxin selectively antagonizes RORγt transcriptional activity.

METHODUsing molecular docking in combination with molecular electrostatic potential (MEP), we detected the interaction between the derivative of digoxin (Dhd) and ROR transcription factor (RORα,RORβ and RORγt), and the results were further confirmed by bioluminescent assay.

RESULTMolecular docking demonstrated that Dhd could exclusively inhibit the conformation of RORγt; bioluminescent assay further indicated that RORγt was selectively antagonized by Dhd in a dose- and time-dependent manner.

CONCLUSIONDhd can selectively suppress RORγt transcriptional activity.

Digoxin ; analogs & derivatives ; pharmacology ; Humans ; Models, Chemical ; Molecular Docking Simulation ; Nuclear Receptor Subfamily 1, Group F, Member 1 ; antagonists & inhibitors ; genetics ; Transcription, Genetic

Objective To study the feasibility and clinical effect of trans umbilical single incision laparoscopic right hemicolectomy for right hemicolon carcinoma.Methods The clinical data of 35 patients with right hemicolon carcinoma were retrospectively analyzed,the 15 cases were received the trans umbilical single incision laparoscopic right hemicolectomy (single incision group) and 20 cases were received laparotomy right hemicolectomy (laparotomy group).The clinical indexes were compared between the 2 groups.Results The length of incision,intraoperative bleeding volume,passage of gas by anus time,feeding time,hospitalized time in single incision group were significantly better than those in laparotomy group [(5.5 ± 0.6) cm vs.(17.6 ± 2.2) cm,(84.0 ± 31.1) ml vs.(155.5 ± 43.1) ml,(2.00 ± 0.76) d vs.(3.75 ± 0.63) d,(5.3 ± 0.6) d vs.(6.5 ± 0.6) d,(9.3 ± 1.4) d vs.(13.5 ± 1.5) d],the operation time in single incision group was significantly longer than that in laparotomy group [(238.4 ± 19.3) min vs.(165.3 ± 25.8) min],there were statistical differences (P ＜ 0.05).There was no statistical difference in number of incision lymph node between the 2 groups (P ＞ 0.05).Postoperative complication in single incision group occurred in 2 cases,postoperative complication in laparotomy group occurred in 3 cases,there was no statistical difference (P ＞0.05).The follow-up time was 1-55 months,the median follow-up time was 28 months,local recurrence and distant metastasis were found in 2 cases in single incision group,and 4 cases was found in laparotomy group,there was no statistical difference (P ＞ 0.05).Conclusions Single incision laparoscopic right hemicolectomy for right hemicolon carcinoma is safe and feasible,which has the advantages of minimal trauma,aesthetic outlook,less bleeding,quick recovery and short hospitalization time,etc.It can be developed in the hospital which has some basis of laparoscopic surgery.

Objetive Psoriasis is an autoimmune-related chronic inflammatory skin disease strongly associated with the dysfunction of Th17 cells. Retinoic acid-related orphan nuclear receptorγt (RORγt) plays a critical role in the differentiation and maturation of Th17 cells and in cell-derived immunologic derangement. We conducted this study to investigate potential mechanism by which the derivative of digoxin selectively antagonizes RORγt transcriptional activity. Method Using molecular docking in combination with molecular electrostatic potential (MEP), we detected the interaction between the derivative of digoxin (Dhd) and ROR transcription factor (RORα,RORβ and RORγt), and the results were further confirmed by bioluminescent assay. Result Molecular docking demonstrated that Dhd could exclusively inhibit the conformation of RORγt;bioluminescent assay further indicated that RORγt was selectively antagonized by Dhd in a dose- and time-dependent manner. Conclusion Dhd can selectively suppress RORγt transcriptional activity.

Objetive Psoriasis is an autoimmune-related chronic inflammatory skin disease strongly associated with the dysfunction of Th17 cells. Retinoic acid-related orphan nuclear receptorγt (RORγt) plays a critical role in the differentiation and maturation of Th17 cells and in cell-derived immunologic derangement. We conducted this study to investigate potential mechanism by which the derivative of digoxin selectively antagonizes RORγt transcriptional activity. Method Using molecular docking in combination with molecular electrostatic potential (MEP), we detected the interaction between the derivative of digoxin (Dhd) and ROR transcription factor (RORα,RORβ and RORγt), and the results were further confirmed by bioluminescent assay. Result Molecular docking demonstrated that Dhd could exclusively inhibit the conformation of RORγt;bioluminescent assay further indicated that RORγt was selectively antagonized by Dhd in a dose- and time-dependent manner. Conclusion Dhd can selectively suppress RORγt transcriptional activity.

Objective To investigate the extracellular polysaccharide distribution and components of Ureaplasma urealyticum (Uu) after biofilm having been developed in.Methods The standard serotype 3 and serotype 14 belong to biovar Parvo,and the standard serotype 4 and serotype 8 belong to biovar T960 were employed to form biofilrns in vitro.Scanning electron microscope and confocal laser scanning microscope were used to analysis the biofilms and extracellular polysaccharide.We used combination of two different labeled lectins,Canavalia ensiformis(FITC-ConA) and Erythrina cristagalli(ECA) which bind to specific polysaccharide residues to visualize extracellular polysaccharide in biofilms,and average uorescence intensity was evaluated Results All the strains can form the biofilmsin vitro.The biofilm was honeycomb-Like structures mainly,and extracellular polymeric substances accounts for majority of proportions.All the extracellular polysaccharide could be combined with FITC-ConA and ECA,and the total average fluorescence intensity of FITC-ConA was higher than ECA( P＜0.001 ).Conclusion Ureaplasma urealyticum biofilm is honeycomb-like structures mainly.The extracellular polysaccharide contains,galactose,and N-acetyl glucan residual,and the glucose,mannose residual are the main components.

Objective To investigate the role of cathepsin G in photoaged fibroblasts. Methods Human fibroblasts were cultured and induced to premature senescence using UVA + MOP methods. Senescence-associated-β-galactosidase (SA-β-gal) stain was used to evaluate the positive rate of aged cells. The mRNA and protein expression of cathepsin G in photoaged fibroblasts were detected by real-time RT-PCR and Western blot techniques. Results Over 98 % induced cells presented a positive SA-β-gal straining. The expression of cathepsin G, detected by Western blot, was increased to (1. 70±0. 028) times of the control. And RT-PCR revealed that the synthesis of cathepsin G mRNA was also up-regulated to 1. 42±0. 09. Conclusion The results of our study demonstrates a significant correlation between photoaged fibroblasts and cathepsin G. The up-regulation of cathepsin G may play an important role in the damages of extracellular matrix and activation of MMPS in photoaged human skin.