Objective To distinguish Cremastra appendiculata(D.Don)Makino,Pleione yunnanensis Rolfe and Pleione bulbocodioides,and its easily confusing products Oreorchis patens and Iphigenia indica Kunth using the ITS sequence in DNA barcodes;To explore the genetic diversity of Cremastra appendiculata germplasm resources.Methods Three different original Cremastra appendiculata,Pleione yunnanensis Rolfe and Pleione bulbocodioides,and their easily confusing products Cremastrae Pseudobulbus of Oreorchis patens and Iphigenia indica Kunth were selected as the research objects,and the genomic DNA of the above samples were extracted by the modified CTAB method,and then the ITS sequences were amplified,sequenced and spliced by PCR technology.The Kimura 2-Parameter(K2P)model was used to calculate the genetic distance,and the phylogenetic tree was constructed with the help of neighbour joining method(NJ)for genetic relationship analysis.Results Except for the Iphigenia indica Kunth species that were not found during the BLAST search,the BLAST comparison results of the other samples were higher than 95%.At the same time,the results of phylogenetic tree showed that Cremastra appendiculata,Pleione yunnanensis Rolfe and Pleione bulbocodioides were clustered into one branch,respectively,and the easily confusing products were also respectively clustered into one branch.Conclusion The ITS sequence in DNA barcodes can be used to accurately distinguish Cremastra appendiculata,Pleione yunnanensis Rolfe and Pleione bulbocodioides,and its easily confusing products Oreorchis patens and Iphigenia indica Kunth.

Objective:To evaluate the feasibility of a novel flexible ultrasound imaging catheter for high-resolution real-time imaging of deep abdominal structures via percutaneous intervention.Methods:A self-developed flexible ultrasound catheter system（outer diameter：3.3 mm，integrated with a 10.5 MHz high-frequency linear array transducer and 4-degree-of-freedom active deflection）was used to image abdominal organs（liver，gallbladder，pancreas，etc.）in two healthy beagles via percutaneous puncture，with comparison to conventional surface convex array probes（3.6 MHz）. Image quality was assessed using a double-blind design，independently scored by five ultrasound physicians with over 10 years of experience on a 6-point scale. Mixed-effects ordinal logistic regression（adjusted for dog，organ，and evaluator variability）was employed for inter-group comparisons，with effect sizes quantified by Cliff's delta. Characteristic anatomical structure detection rates were analyzed using generalized linear mixed models（Firth correction for zero events）. Inter-rater reliability was assessed via intraclass correlation coefficient（ICC，two-way random model）and Kendall's coefficient of concordance.Results:The flexible catheter group demonstrated significantly superior image quality compared to the surface probe group . The most pronounced improvement was observed in adrenal imaging（5 points vs. 1 point， P<0.001，δ=0.92），while pancreatic imaging showed no significant difference（2 points vs. 2 points， P=0.812，δ=0.05）. Other organs exhibited significant quality enhancements（all P<0.05，δ=0.63～0.81）. The catheter significantly improved detection rates of characteristic structures：gastric wall stratification（90% vs. 10%， OR=36.0， P<0.001），adrenal corticomedullary differentiation（80% vs. 0， OR=∞， P<0.001），and gallbladder/small intestinal wall trilaminar structure（80% vs. 20%， OR=12.0， P=0.003）. The high-quality image rate（≥5 points）reached 77.8%，representing an 11.6-fold improvement over surface ultrasound（6.7%， P<0.001）. Inter-rater reliability was excellent（ICC=0.85，W=0.79）. Conclusions:Percutaneous flexible ultrasound catheter imaging significantly enhances resolution and anatomical detail visualization of deep abdominal structures. Combining minimally invasive access with operational flexibility，this technology shows promise for providing precise image guidance in minimally invasive procedures.

Objective To distinguish Cremastra appendiculata(D.Don)Makino,Pleione yunnanensis Rolfe and Pleione bulbocodioides,and its easily confusing products Oreorchis patens and Iphigenia indica Kunth using the ITS sequence in DNA barcodes;To explore the genetic diversity of Cremastra appendiculata germplasm resources.Methods Three different original Cremastra appendiculata,Pleione yunnanensis Rolfe and Pleione bulbocodioides,and their easily confusing products Cremastrae Pseudobulbus of Oreorchis patens and Iphigenia indica Kunth were selected as the research objects,and the genomic DNA of the above samples were extracted by the modified CTAB method,and then the ITS sequences were amplified,sequenced and spliced by PCR technology.The Kimura 2-Parameter(K2P)model was used to calculate the genetic distance,and the phylogenetic tree was constructed with the help of neighbour joining method(NJ)for genetic relationship analysis.Results Except for the Iphigenia indica Kunth species that were not found during the BLAST search,the BLAST comparison results of the other samples were higher than 95%.At the same time,the results of phylogenetic tree showed that Cremastra appendiculata,Pleione yunnanensis Rolfe and Pleione bulbocodioides were clustered into one branch,respectively,and the easily confusing products were also respectively clustered into one branch.Conclusion The ITS sequence in DNA barcodes can be used to accurately distinguish Cremastra appendiculata,Pleione yunnanensis Rolfe and Pleione bulbocodioides,and its easily confusing products Oreorchis patens and Iphigenia indica Kunth.

Objective:To evaluate the feasibility of a novel flexible ultrasound imaging catheter for high-resolution real-time imaging of deep abdominal structures via percutaneous intervention.Methods:A self-developed flexible ultrasound catheter system（outer diameter：3.3 mm，integrated with a 10.5 MHz high-frequency linear array transducer and 4-degree-of-freedom active deflection）was used to image abdominal organs（liver，gallbladder，pancreas，etc.）in two healthy beagles via percutaneous puncture，with comparison to conventional surface convex array probes（3.6 MHz）. Image quality was assessed using a double-blind design，independently scored by five ultrasound physicians with over 10 years of experience on a 6-point scale. Mixed-effects ordinal logistic regression（adjusted for dog，organ，and evaluator variability）was employed for inter-group comparisons，with effect sizes quantified by Cliff's delta. Characteristic anatomical structure detection rates were analyzed using generalized linear mixed models（Firth correction for zero events）. Inter-rater reliability was assessed via intraclass correlation coefficient（ICC，two-way random model）and Kendall's coefficient of concordance.Results:The flexible catheter group demonstrated significantly superior image quality compared to the surface probe group . The most pronounced improvement was observed in adrenal imaging（5 points vs. 1 point， P<0.001，δ=0.92），while pancreatic imaging showed no significant difference（2 points vs. 2 points， P=0.812，δ=0.05）. Other organs exhibited significant quality enhancements（all P<0.05，δ=0.63～0.81）. The catheter significantly improved detection rates of characteristic structures：gastric wall stratification（90% vs. 10%， OR=36.0， P<0.001），adrenal corticomedullary differentiation（80% vs. 0， OR=∞， P<0.001），and gallbladder/small intestinal wall trilaminar structure（80% vs. 20%， OR=12.0， P=0.003）. The high-quality image rate（≥5 points）reached 77.8%，representing an 11.6-fold improvement over surface ultrasound（6.7%， P<0.001）. Inter-rater reliability was excellent（ICC=0.85，W=0.79）. Conclusions:Percutaneous flexible ultrasound catheter imaging significantly enhances resolution and anatomical detail visualization of deep abdominal structures. Combining minimally invasive access with operational flexibility，this technology shows promise for providing precise image guidance in minimally invasive procedures.

Objective:To investigate the etiological characteristics of hand, foot and mouth disease (HFMD) and the VP1 gene evolution of coxsackievirus A6 (CVA6) in Qingdao from 2020 to 2022.Methods:Total enterovirus (EV), enterovirus 71 (EV71), coxsackievirus A16 (CVA16), CVA6 and coxsackievirus A10 (CVA10) were detected by fluorescent RT-PCR in throat swab samples from HFMD patients in Qingdao from 2020 to 2022. The full-length sequence amplification of VP1 gene and gene sequence determination of CVA6 isolates were performed, and the phylogenetic and molecular characteristics were analyzed by MEGA-X software.Results:The overall positive rate of enteroviruses in 1 645 samples collected during 2020-2021 was 80.73%(1 328/1 645). The proportions of EV71, CVA16, CVA6, CVA10 and other enteroviruses were 0.08%(1/1 328), 19.65%(261/1 328), 68.15%(905/1 328), 0.98%(13/1 328) and 11.14%(148/1 328), respectively. Children under 5 years old accounted for 75.68% (1 245/1 645) of all cases and 100% (2/2) of severe cases. A phylogenetic tree was constructed based on the VP1 region sequences of 45 CVA6 strains showing all strains belonging to the D3a subtype. There were 10 non-synonymous mutations of amino acids in the VP1 protein by gene evolution analysis.Conclusions:CVA6 D3a subtype is the main epidemic strain causing HFMD in Qingdao from 2020 to 2022, especially in children under 5 years old.

OBJECTIVE To investigate the intervention effect of kushenol F （KSC-F） on ulcerative colitis （UC） mice. METHODS Totally 30 male C57BL/6J mice were randomly divided into the normal group， model group， positive drug group （sulfasalazine， 703 mg/kg）， KSC-F 50 mg/kg group （KSC-F50 group）， and KSC-F 100 mg/kg group （KSC-F100 group）， with 6 mice in each group. Except for the normal group， the mice in the remaining groups were given 3% dextran sulfate sodium solution continuously for 7 days to induce UC model. Concurrently， administration groups received corresponding drug solution intragastrically， once a day， for 10 consecutive days. During the experiment， the changes in body weight and bowel movements of the mice were observed. Disease activity index scoring was performed after the last administration. The histopathological morphology of colonic tissue was examined. The levels of inflammatory factors in the serum and colon tissue were measured. Additionally， the mRNA expression of inflammatory factors， and the protein expressions of inflammation-related proteins [interleukin-1β （IL-1β）， forkhead box O1（FOXO1）， phosphoinositide 3-kinase（PI3K）， phosphorylated PI3K（p-PI3K）， p38 mitogen-activated protein kinase（p38 MAPK）， phosphorylated p38 MAPK（p-p38 MPAK） and phosphorylated protein kinase B（p- Akt）] were determined in colonic tissue. RESULTS KSC-F could alleviate weight loss and colonic tissue damage in UC mice. KSC- F reduced the levels of IL-1β， IL-6， IL-8 and tumor necrosis factor-α （TNF-α） in serum， as well as IL-1β， IL-6， IL-17 and TNF- α in colonic tissue to varying degrees and increased the levels of IL-10 in both serum and colonic tissue （P＜0.05 or P＜0.01）. Moreover， KSC-F decreased the expression levels of IL-1β， IL-17 and TNF-α mRNA， as well as p-PI3K， p-p38 MAPK， and p- Akt proteins in colonic tissue to varying degrees， and increased the expression levels of IL-10 mRNA and FOXO1 protein in colonic tissue （P＜0.05 or P＜0.01）. CONCLUSIONS KSC-F effectively alleviates UC symptoms in mice by inhibiting PI3K， Akt and p38 MAPK activation， mitigating the release of pro-inflammatory factors such as IL-1β， IL-6， TNF- α，promoting the anti-inflammatory factor IL-10 secretion， and reducing inflammation-induced colonic tissue damage.

Both natural ginsenoside F2 and unnatural ginsenoside 3β,20S-Di-O-Glc-DM were reported to exhibit anti-tumor activity. Traditional approaches for producing them rely on direct extraction from Panax ginseng, enzymatic catalysis or chemical synthesis, all of which result in low yield and high cost. Metabolic engineering of microbes has been recognized as a green and sustainable biotechnology to produce natural and unnatural products. Hence we engineered the complete biosynthetic pathways of F2 and 3β,20S-Di-O-Glc-DM in Saccharomyces cerevisiae via the CRISPR/Cas9 system. The titers of F2 and 3β,20S-Di-O-Glc-DM were increased from 1.2 to 21.0 mg/L and from 82.0 to 346.1 mg/L at shake flask level, respectively, by multistep metabolic engineering strategies. Additionally, pharmacological evaluation showed that both F2 and 3β,20S-Di-O-Glc-DM exhibited anti-pancreatic cancer activity and the activity of 3β,20S-Di-O-Glc-DM was even better. Furthermore, the titer of 3β,20S-Di-O-Glc-DM reached 2.6 g/L by fed-batch fermentation in a 3 L bioreactor. To our knowledge, this is the first report on demonstrating the anti-pancreatic cancer activity of F2 and 3β,20S-Di-O-Glc-DM, and achieving their de novo biosynthesis by the engineered yeasts. Our work presents an alternative approach to produce F2 and 3β,20S-Di-O-Glc-DM from renewable biomass, which lays a foundation for drug research and development.

There are direct and indirect interactions between gut microbiota and host immune response, which can have a multifaceted impact on host health. Dysbiosis caused by disturbances in the gut microbiota is associated with susceptibility to many diseases, especially immune-related diseases. Based on the research results in recent years, this paper introduced the mechanism of the interaction between gut microbiota and host immunity, and expounded the role of gut microbiota in the occurrence and development of immune-related diseases, including intestinal system diseases such as inflammatory bowel disease and other systemic diseases such as rheumatoid arthritis, and summarized disease treatment strategies targeting gut microbiota. A better understanding of the research progress of gut microbiota and immune-related diseases will help us in the prevention and management of such diseases, and broaden our path to discover disease intervention targets.
Gastrointestinal Microbiome/physiology* ; Humans ; Inflammatory Bowel Diseases ; Intestines

Objective:To investigate the changes of serum catecholamines (CST) and oxidative stress index in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) with hypertension and the correlation with blood pressure.Methods:Total of 100 OSAHS patients admitted to Jingmen No.1 People′s Hospital from February 2018 to August 2020 were selected. According to whether patients combined with hypertension, they were divided into OSAHS group (54 cases) and OSAHS combined with hypertension group (46 cases). The changes of serum CST and oxidative stress index in the two groups were compared and analyzed the correlation of blood pressure and CST and oxidative stress index.Results:The level of myeloperoxidase(MPO) in the OSAHS combined with hypertension group was higher than that in the OSAHS group: (41.67 ± 3.02) U/L vs. (39.11 ± 2.65) U/L; the levels of catalase (CAT) and CST in the OSAHS combined with hypertension group were higher than those in the OSAHS group: (11.47 ± 2.54) kU/L vs. (15.88 ± 2.31) kU/L, (23.63 ± 4.38) ng/L vs. (28.61 ± 5.42) ng/L, there were statistically differences ( P<0.05). The levels of respiratory disturbance index, lowest oxygen saturation, triacylglycerol, total cholesterol, low density lipoprotein cholesterin, high density lipoproteincholesterol, epinephrine, noradrenaline in the two groups had no significant differences ( P>0.05). Spearman correlation analysis showed that the systolic pressure and diastolic pressure had positive correlation with MPO ( P<0.05), and had negative correlation with CAT and CST ( P<0.05). Conclusions:The serum CST and CAT are down-regulated in patients with OSAHS with hypertension, and serum MPO is up-regulated. Oxidative stress may be related to OSAHS with hypertension.

Triterpenoid saponins are widely used in medicine, health cares, cosmetics, food additives and agriculture because of their unique chemical properties and rich pharmacological activities. UDP-dependent glycosyltransferases (UGTs) are the key enzymes involved in triterpenoid saponin biosynthesis, and play important roles in the diversity of triterpenoid saponin structures and pharmacological activities. This review summarized the UGTs involved in plant triterpenoid saponin biosynthesis based on the sources of UGTs and the types of receptors. Moreover, the application of UGTs in heterologous biosynthesis of triterpenoid saponins based on synthetic biology was also discussed.
Glycosyltransferases/genetics* ; Plants ; Saponins/chemistry* ; Triterpenes