A method was developed to elucidate the structures of sulfated oligosaccharides through establishment of an effective reversed phase ion pair ultra-performance liquid chromatography coupled with electrospray ionization time of flight mass spectrometry( RPIP-UPLC-ESI-TOF-MS). Heptylamine (20 mmol/L,pH4) has been selected as the ion-pairing agent.κ-Carrageenan oligosaccharides have been separated on BEH C_(18) column using MeOH/H_2O with 25% heptylammonium formate as eluent in linear gradient mode. Mass spectra were obtained by ESI-Q-TOF-MS in both positive and negative modes. κ-Carrageenan oligosaccharides were well separated up to pentatetrasaccharide,and ESIMS analysis for κ-carrageenan oligosaccharides up to hepta-cosasccharide. The results showed that all acid hydrolyzed κ-carrageenan oligosaccharides were odd sugars,which was further confirmed by polyacrylamide gel electrophoresis ( PAGE). The characteristic fragmentation pattern of ion-pair oligosaccharides in mass spectra can be applied for rapid structure identification.

Objective To investigate the therapeutic effects of recombinant yeast hepatitis B virus(HBV)vaccine combined with interferon(IFN)α-1b and determine the rational dosage of HBV vaccine for the further clinical study with larger sample.Methods Two hundreds and sixteen patients with chronic hepatitis B(CHB)were enrolled in this randomized,multi-center,double-blinded and placebo-controlled clinical trial.All the subjects were not treated with antiviral drugs within 6 months and randomly divided into 90μg,60μg and placebo groups with a ratio of 1:1:1.All the patients were intramuscularly administrated with 90μg or 60μg recombinant HBV vaccine or placebo at week 0,2,6,10,14,18,22,respectively.Meanwhile,they were also treated with IFNα-1b 50μg,3 times a wcek for 24 weeks.All patients were followed up for 24 weeks after withdrawal of anti-HBV therapy.Serum HBV DNA level,HBeAg titer and liver function were monitored frequently.Interferon-γ secreting lymphoeytes were detected by Enzyme-linked immunospot(ELISPOT)in part of the patients.Results The serum HBV DNA levels were(6.03±1.79),(5.52±1.82)and(6.29±1.70)log10 copy/mL at week 24 of treatment in high dose,low dose and placebo groups,respectively (P=0.458).And the serum HBV DNA levels were(5.92±1.98),(5.49±1.99)and(6.16±1.76)log10 copy/mL at weck 24 after withdrawal of treatment,respectively(P=0.720).The rates of patients whose HBV DNA＜1×105 copy/mL in these three groups were 30.4%,39.4% and 20.8% at week 24 of treatment,respectively and there was significant difference between high dose group and low dose group(P=0.015).The rate of patients whose HBV DNA＜1×105 copy/mL at week 24 after withdrawal was highest in low dose group,but no significant differences before and after treatment in these three groups(P=0.257).The HBV DNA negative rates were 17.4%,25.4% and 6.9% in these three groups,respectively,which were significantly different(P=0.012).At week 24 of treatment and week 24 after withdrawal of treatment,the alanine aminotransferase normalization rate,HBeAg seroconversion rate were highest in low dose group,but no significant differences in these three groups.ELISPOT positive rates at week 24 of treatment and week 24 after withdrawal of treatment in high close and low dose groups were higher than that in placebo group(P＜0.05).The incidence of adverse events was similar and there was no drug related severe adverse events in each group.Conclusion Recombinant HBV vaccine maybe contribute to anti-HBV therapy and 60μg of dosage seems to be rational.

OBJECTIVETo analyze the potential risks of fungal contaminants on Panax notoginseng and Amomum tsaoko.

METHODThe primary investigation was conducted in the P. notoginseng and A. tsaoko major production areas in Yunnan. Samples of P. notoginseng and A. tsaoko were collected from drugstores and markets in 3 cities of Yunnan. Dilution-plate method was applied for the isolation of fungi, the obtained species were identified according to morphological and molecular approaches.

RESULTPaecilomyces lilacinus and Penicillium citrinum were dominant on samples of Panax notoginseng. P. lilacinus and Aspergillus flavus were dominant on samples of Amomum tsaoko.

CONCLUSIONIn Yunnan province, the major fungal contaminants on P. notoginseng are P. lilacinus and P. citrinum and the major fungal contaminants on A. tsaoko are P. lilacinus and Aspergillus flavus. There exists a potential contamination risk of citrinin on P. notoginseng and aflatoxin on A. tsaoko.

Amomum ; microbiology ; China ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; Mitosporic Fungi ; isolation & purification ; physiology ; Panax notoginseng ; microbiology ; Risk

Collected and preserved germplasm resource of Dendrobium chrysotoxum to lay the foundation for screening fine germplasm. Through refering literatures, visiting and field survey to investigate the distribution, botanic characters and apply status of D. chrysotoxum, furthermore to collect the germplasm resource. The result show that wild germplasm resource of D. chrysotoxum has obvious differences in stem characters, leaf shape as well as flower color aspects. In addition, in recent ten years, the reserves of D. chrysotoxum germplasm resource seriously descended. Through this study, we can draw a conclusion that D. chrysotoxum germplasm resource exist diversity in biology. In these germplasm resource, there are high yield and good quality variety.
Conservation of Natural Resources ; Dendrobium ; growth & development

Objective To analyze the frequency of pelvis peristalsis in children with hydronephrosis and its correlation with postoperative percentage of improvement in anteroposterior diameter (PI－APD).Methods From Oc-tober 2015 to September 2017,the data of the children with unilateral ureteropelvic junction obstruction (UPJO)hydro-nephrosis who underwent laparoscopic pyeloplasty were collected,and their pelvis peristalsis frequencies of 2 min were recorded after exposing the pelvis,while the APD and PI－APD were followed 1 year posto-peratively.The degrees of hydronephrosis were divided into mild,moderate and severe according to the Society of Fetal Urology(SFU)classifica-tion system.ANOVA was used to compare the pelvis peristalsis frequency and postoperative PI－APD between different degrees of hydronephrosis.Spearman analysis was used to analyze the correlation between pelvis peristalsis frequency and degree of hydronephrosis.Meanwhile,Pearson test was used to analyze the correlation between pelvis peristalsis fre-quency,hydronephrosis degree and PI－APD.Results A total of 50 children were included,of which 36 patients got follow－up for 1 year postoperatively.Pelvis 2 min peristalsis frequency between different degrees of hydronephrosis [mild:(8.1 ± 3. 3)times;moderate:(6.3 ± 3.5)times;severe:(7.8 ± 3.9)times]had no significant difference (F＝0.65,P＝0. 527);no statistical correlation was observed between pelvis peristalsis frequency and degree of hydro-nephrosis or PI－APD (all P ＞0.05).Preoperative hydronephrosis degree was positively correlated with PI －APD (r＝0.54,P＝0. 001).PI－APD in severe hydronephrosis (0.48 ± 0.29)was significantly higher than that of mild hydronephrosis (0.21 ± 0.20)(P ＝0.001 ). Conclusions For children with different degrees of hydronephrosis caused by UPJO,there was no significant difference in the pelvis peristalsis frequency recorded during laparoscopic pyeloplasty.Pelvis peristalsis frequency recorded during surgery was not correlated with PI －APD,but preoperative hydronephrosis degree was positively correlated with postoperative PI－APD.

Isopeptide bond-mediated molecular superglue is the irreversible covalent bond spontaneously formed by the side chains of lysine (Lys) and asparagine/aspartic acid (Asn/Asp) residues. The peptide-peptide interaction is specific, stable, and can be achieved quickly without any particular physicochemical factor. In the light of recent progress by domestic and foreign researchers, here we summarize the origin, assembly system and mechanism of isopeptide bond reaction, as well as the molecular cyclization and protein topological structure mediated by it. The prospect for its application in synthetic vaccine, hydrogel and bacterial nanobiological reactor is further discussed.
Cyclization ; Lysine ; Peptides ; chemistry ; Proteins

The capacity for thermal tolerance is critical for industrial enzyme. In the past decade, great efforts have been made to endow wild-type enzymes with higher catalytic activity or thermostability using gene engineering and protein engineering strategies. In this study, a recently developed SpyTag/SpyCatcher system, mediated by isopeptide bond-ligation, was used to modify a rumen microbiota-derived xylanase XYN11-6 as cyclized and stable enzyme C-XYN11-6. After incubation at 60, 70 or 80 ℃ for 10 min, the residual activities of C-XYN11-6 were 81.53%, 73.98% or 64.41%, which were 1.48, 2.92 or 3.98-fold of linear enzyme L-XYN11-6, respectively. After exposure to 60-90°C for 10 min, the C-XYN11-6 remained as soluble in suspension, while L-XYN11-6 showed severely aggregation. Intrinsic and 8-anilino-1-naphthalenesulfonic acid (ANS)-binding fluorescence analysis revealed that C-XYN11-6 was more capable of maintaining its conformation during heat challenge, compared with L-XYN11-6. Interestingly, molecular cyclization also conferred C-XYN11-6 with improved resilience to 0.1-50 mmol/L Ca²⁺ or 0.1 mmol/L Cu²⁺ treatment. In summary, we generated a thermal- and ion-stable cyclized enzyme using SpyTag/SpyCatcher system, which will be of particular interest in engineering of enzymes for industrial application.
Animals ; Cyclization ; Endo-1,4-beta Xylanases ; chemistry ; metabolism ; Enzyme Stability ; Industrial Microbiology ; methods ; Microbiota ; Protein Engineering ; Rumen ; enzymology ; microbiology ; Temperature

BackgroundSchizophrenic patients have metabolic disorders， impaired language and information processing function. Inflammatory factors may play an important role in the occurrence and development of schizophrenia. ObjectiveTo explore the relationship of the inflammatory factor levels with metabolic levels， language fluency and information processing function in patients with schizophrenia， so as to provide references for clinical understanding of the neuropathological mechanisms of schizophrenia. MethodsA total of 96 patients with schizophrenia were included in the study group， who were hospitalized in the Fourth People's Hospital of Hefei from January 2021 to December 2022 as well as met the diagnostic criteria of Diagnostic and Statistical Manual of Mental Disorders， fifth edition （DSM-5） and Mini-International Neuropsychiatric Interview （MINI） 6.0 .Meanwhile， population who underwent physical examination at the same hospital were included in the control group （n=42）. A high-sensitivity multi factor electrochemiluminescence analyzer was used to detect the levels of inflammatory factors IL-4， IL-5， IL-7， IL-8， IL-10 and IL-13. A fully automated biochemical analyzer was used to detect the levels of metabolic indicators such as fasting blood glucose， triglycerides， high-density lipoprotein， apolipoprotein A， creatinine and urea nitrogen. Verbal fluency and information processing function of all participants were assessed by using Verbal Fluency Test （VFT） and Stroop Color Word Test （SCWT）. ResultsThere were statistically significant differences in the levels of IL-4， IL-5， IL-7， IL-8， IL-10， IL-13 and IL-15 between the study group and the control group （P<0.05）. There were statistically significant differences in BMI， waist circumference， fasting blood glucose， triglycerides， high-density lipoprotein， urea nitrogen， apolipoprotein A and creatinine levels between the two groups （P<0.05）. The differences in the correct number of household appliances， animals， fruits， vegetables， names starting with "water" and "self" in VFT between the two groups were statistically significant （P<0.05）. The differences in point reaction time， character reaction time and character color reaction time in SCWT between the two groups were statistically significant （P<0.05）. Correlation analysis showed that except for creatinine levels， the levels of IL-4 and IL-5 in patients with schizophrenia were correlated with other indicators （P<0.05）. IL-7 levels were correlated with creatinine levels， household appliances， animals， fruits， correct number of names starting with "water" in VFT， point reaction time and word reaction time in SCWT （P<0.05）. IL-8 levels were correlated with triglyceride levels， household appliances， animals， fruits， vegetables， correct number of names starting with "water" and "self" in VFT and word reaction time in SCWT （P<0.05）. Except for creatinine levels and the correct number of names starting with "self"， IL-10 levels were correlated with all other indicators （P<0.05）. Except for creatinine and urea nitrogen levels， IL-13 levels were correlated with other indicators （P<0.05）. ConclusionThe levels of inflammatory factors in patients with schizophrenia may be related to their metabolic levels， language fluency and information processing function. ［Funded by Anhui Province Key Research and Development Plan Project （number， 2022e07020002）］