ObjectiveTo investigate the immunomodulatory effect of polysaccharides from Astragali Radix-Angelicae Sinensis Radix herb pair（Qi-gui polysaccharides） on lipopolysaccharide（LPS）-induced RAW264.7 macrophages and to characterize the structure of the active component Qi-gui homogeneous polysaccharide（AAPS-4a）， and evaluate its protective effect on ulcerative colitis（UC）. MethodsThe effects of six Qi-gui polysaccharides（0.01-100 mg·L^-1） on the proliferation of RAW264.7 cells were assessed by cell proliferation and activity assay（CCK-8）， and enzyme-linked immunosorbent assay（ELISA） was used to investigate the effects of the six polysaccharides（3， 10 mg·L^-1） on the secretion levels of tumor necrosis factor（TNF）-α， interferon（IFN）-β， and nitric oxide（NO） in LPS-induced RAW264.7 cells. After screening for active polysaccharides， high-performance size-exclusion chromatography（HPSEC） was used to determine its homogeneity and relative molecular weight， then its characteristic functional groups were identified by Fourier transform infrared spectroscopy（FT-IR）， monosaccharide composition was analyzed by high performance liquid chromatography（HPLC）， methylation analysis combined with gas chromatography-mass spectrometry（GC-MS） was performed to determine the types and linkage modes of sugar residues， and one- and two-dimensional nuclear magnetic resonance（NMR） were used to identify the sugar residue composition and configuration of the active polysaccharide. Finally， experimental animals were divided into the normal group， model group， AAPS-4a low-dose group（50 mg·kg^-1）， AAPS-4a high-dose group（100 mg·kg^-1）， and sulfasalazine（SASP） group （75 mg·kg^-1）. Except for the normal group， the acute UC mouse model was induced using 3.5% dextran sulfate sodium salt（DSS）. Each treatment group was administered the corresponding dose via oral gavage for 7 days， and changes in body weight were recorded. After treatment， the spleen index and disease activity index（DAI） score were calculated， TNF-α and interleukin-6（IL-6） levels in the serum were detected by ELISA， and histopathological changes in colon tissue were observed by hematoxylin-eosin（HE） staining. ResultsAt the cellular level， AAPS-4a exhibited a dose-dependent inhibition of LPS-induced increases in TNF-α， IFN-β， and NO levels（P<0.01）. Structural characterization of AAPS-4a revealed that it was a homogeneous polysaccharide with a relative molecular weight of 7.6×10³ Da， consisting of mannose（Man）， glucose（Glc）， and galactose（Gal） in a molar ratio of 1.3∶23.9∶1.0. It was primarily composed of five sugar residues of 1，6-α-D-Glcp， T-α-D-Glcp， 1，3-β-D-Galp， 1，4-α-D-Manp， and 1，2-α-D-Galp. In vivo experiments showed that compared with the normal group， the model group demonstrated markedly increased DAI score and spleen index， significantly reduced colon length， and significantly elevated levels of TNF-α and IL-6（P<0.01）. Compared with the model group， the AAPS-4a high-dose group significantly reduced the DAI score and spleen index， as well as TNF-α and IL-6 levels， and improved colonic atrophy（P<0.05， P<0.01）. Pathological observations showed that AAPS-4a significantly inhibited inflammatory cell infiltration in colon tissue and alleviated pathological damage. ConclusionAAPS-4a， a neutral homogeneous polysaccharide composed of 1，6-α-D-Glcp， T-α-D-Glcp， 1，3-β-D-Galp， 1，4-α-D-Manp and 1，2-α-D-Galp， is identified as a key bioactive component contributing to the anti-UC effect of the Qi-gui herb pair. Its immunoregulatory and anti-UC properties suggest its potential as a therapeutic agent for UC.

OBJECTIVE To investigate the stability of 5-fluorouracil （5-FU） in human blood and to establish a standardized clinical sampling and delivery procedure for therapeutic drug monitoring （TDM） of 5-FU. METHODS The EDTA-anticoagulated whole blood was used as the matrix to prepare stability assessment samples of 5-FU at both low （200 ng/mL） and high （5 000 ng/mL） concentrations （with groups without stabilizer and with 1% volume ratio of stabilizer）. The stability assessment samples were placed under room temperature （[ 25±2） ℃] and refrigerated （2-8 ℃） conditions， with sampling at 0， 0.5， 1， 2， 4， 7， and 24 h. After vortexing and centrifugation， the upper plasma layer was collected； proteins were precipitated using methanol， and the concentration of 5-FU in plasma was determined by liquid chromatography-tandem mass spectrometry. Based on the whole blood stability results， clinical sampling and delivery procedures were established. RESULTS The concentration of 5-FU in blank whole blood samples without stabilizers was significantly lower than that in samples with stabilizers （P＜0.05）. However， varying volumes （10， 25， 50 μL） of stabilizers had no significant effect on the measured concentrations of 5-FU in stability assessment samples with low and high concentrations （P＞0.05）. Without the addition of a stabilizer， low- and high-concentration 5-FU whole blood samples remained stable at room temperature for 0.5 h and 1 h， respectively， and under refrigeration for 2 h and 7 h， respectively. After the addition of a 1% stabilizer， the whole blood samples remained stable for up to 24 h under both room temperature and refrigerated conditions. Based on these findings， the following procedure was established： after collection， whole blood samples could be temporarily stored at room temperature （≤0.5 h） or at 4　℃ （≤2 h）， and transported at 2-8　℃. Upon delivery to the laboratory， a 1% volume ratio of stabilizer must be added immediately， followed by centrifugation within 24 h. The resulting plasma should be stored at －20 ℃ . CONCLUSIONS 5-FU in whole blood exhibits poor stability at room temperature. Refrigeration at 2-8　℃ slightly improves stability ， but degradation still occurs rapidly. Adding a stabilizer at a 1% volume ratio significantly prolongs the refrigerated storage time. The established sampling and transport procedure for 5-FU TDM innovatively introduces the stabilizer addition step at the laboratory sample reception stage （rather than immediately after blood draw）. This approach ensures analytical quality while offering greater adaptability to real-world clinical sampling conditions， significantly improving practical feasibility.

To screen and identify a chitosanase with high stability, we cloned the chitosanase gene from Bacillus atrophaeus with a high protease yield from the barren saline-alkali soil and expressed this gene in Escherichia coli. The expressed chitosanase of B. atrophaeus (BA-CSN) was purified by nickel-affinity column chromatography. The properties including optimal temperature, optimal pH, substrate specificity, and kinetic parameters of BA-CSN were characterized. The results showed that BA-CSN had the molecular weight of 31.13 kDa, the optimal temperature of 55 ℃, the optimal pH 5.5, and good stability at temperatures below 45 ℃ and pH 4.0-9.0. BA-CSN also had good stability within 4 h of pH 3.0 and 10.0, be activated by K⁺, Na⁺, Mn²⁺, Ca²⁺, Mg²⁺, and Co²⁺, (especially by Mn²⁺), and be inhibited by Fe³⁺, Cu²⁺, and Ag⁺. BA-CSN showcased the highest relative activity in the hydrolysis of colloidal chitosan, and it had good hydrolysis ability for colloidal chitin. Under the optimal catalytic conditions, BA-CSN demonstrated the Michaelis constant K_m and maximum reaction rate V_max of 9.94 mg/mL and 26.624 μmoL/(mL·min), respectively, for colloidal chitosan. In short, BA-CSN has strong tolerance to acids and alkali, possessing broad industrial application prospects.
Bacillus/genetics* ; Hydrogen-Ion Concentration ; Escherichia coli/metabolism* ; Glycoside Hydrolases/biosynthesis* ; Substrate Specificity ; Enzyme Stability ; Chitosan/metabolism* ; Temperature ; Kinetics ; Cloning, Molecular ; Bacterial Proteins/biosynthesis* ; Recombinant Proteins/genetics*

Acute-on-chronic liver failure （ACLF） refers to severe liver dysfunction that occurs on the basis of chronic liver diseases， and it is characterized by rapid disease progression， poor prognosis， and high mortality rate. In recent years， inflammatory markers have become a research hotspot due to their significant role in assessing the prognosis of ACLF. This article reviews the advances in the application of inflammatory markers in assessing the prognosis of ACLF， such as systemic immune inflammatory index， lymphocyte-white blood cell ratio， neutrophil-lymphocyte ratio， and C-reactive protein， and discusses their clinical value and future research directions， in order to provide a theoretical basis for the early intervention and prognosis management of ACLF patients.

Environmental pollution is closely linked to the occurrence and development of cancer. Chemical carcinogens are the most important environmental factors causing cancer in humans. Among them, persistent organic pollutants (POPs) are characterized by their widespread distribution, persistence, and bioaccumulation. Research on the carcinogenic effects of POPs has received considerable attention in recent years. This article reviewed the internal exposure, association with cancer risk, and potential carcinogenic mechanisms of five typical classes of POPs in the environment, including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), per- and poly-fluoroalkyl substances (PFAS), brominated flame retardants (BFRs), and short-chain chlorinated paraffins (SCCPs). These five types of POPs have distinct carcinogenic mechanisms, including interfering with cell proliferation cycle, altering epigenetic inheritance, promoting oxidative stress, altering energy metabolism, and affecting immune function. The development of cancer is the result of interaction between intrinsic genetic factors and external environmental factors. In addition to focusing on how environmental POPs affect the genetic material of organisms, it is also important to consider their effects on the tumor microenvironment, including tumor immunity and angiogenesis. Understanding these effects is crucial for guiding future efforts in pollution control and precision medicine in cancer treatment.

Objective To evaluate the stability and aesthetic effect of a xenogeneic collagen matrix(mucograft)on achieving an adequate keratinized mucosa width(KMW)around implants and to provide a reference basis for the clini-cal application of xenogeneic collagen matrix materials.Methods The hospital ethics committee approved the study protocol,and the patients provided informed consent.Twenty patients with a KMW＜2 mm at the buccal implant site who were treated in Binzhou Medical University Affiliated Yantai Stomatological Hospital from July 2020 to September 2022 were included,and a total of 36 implants were included.The mean age of the patients was(52.0±10.4)years,of which 18 were females and 2 were males.They were divided into a free gingival graft group(FGG,control group)and a xenogeneic collagen matrix group(test group)according to different graft materials.The incremental effect of the KMW on the buccal side of the implant and the mucosal shrinkage rate was measured at 1 month and 3 months after the opera-tion.The mucosal scar index(MSI)was evaluated after the operation.Results At 3 months postoperatively,the KMW was(3.67±1.06)mm in the control group and(2.96±0.98)mm in the test group,and the difference was statistically significant(t=2.076,P＜0.05).The KMW shrinkage rate was(33.34±16.30)％in the test group and(22.05± 15.47)％in the control group at 1 month postoperatively and(51.95±12.60)％in the test group and(37.44±16.30)％in the control group at 3 months postoperatively,with statistically significant differences between the two groups at the same time points(P＜0.05).Three months after surgery,the test group showed significantly better outcomes than the control group in terms of the five scar indicators(scar width,scar convexity,scar color,scar trace,and overall appear-ance),and the difference was statistically significant(P＜0.05).Conclusion Xenogeneic collagen matrix can increase the peri-implant KMW and achieve a more natural and coordinated soft tissue aesthetic effect but with a higher shrink-age rate.

Digital intraoral scanning is a hot topic in the field of oral digital technology.In recent years,digital intra-oral scanning has gradually become the mainstream technology in orthodontics,prosthodontics,and implant dentistry.The precision of digital intraoral scanning and the accuracy and stitching of data collection are the keys to the success of the impression.However,the operators are less familiar with the intraoral scanning characteristics,imaging process-ing,operator scanning method,oral tissue specificity of the scanned object,and restoration design.Thus far,no unified standard and consensus on digital intraoral scanning technology has been achieved at home or abroad.To deal with the problems encountered in oral scanning and improve the quality of digital scanning,we collected common expert opin-ions and sought to expound the causes of scanning errors and countermeasures by summarizing the existing evidence.We also describe the scanning strategies under different oral impression requirements.The expert consensus is that due to various factors affecting the accuracy of digital intraoral scanning and the reproducibility of scanned images,adopting the correct scanning trajectory can shorten clinical operation time and improve scanning accuracy.The scanning trajec-tories mainly include the E-shaped,segmented,and S-shaped methods.When performing fixed denture restoration,it is recommended to first scan the abutment and adjacent teeth.When performing fixed denture restoration,it is recommend-ed to scan the abutment and adjacent teeth first.Then the cavity in the abutment area is excavated.Lastly,the cavity gap was scanned after completing the abutment preparation.This method not only meets clinical needs but also achieves the most reliable accuracy.When performing full denture restoration in edentulous jaws,setting markers on the mucosal tissue at the bottom of the alveolar ridge,simultaneously capturing images of the vestibular area,using different types of scanning paths such as Z-shaped,S-shaped,buccal-palatal and palatal-buccal pathways,segmented scanning of dental arches,and other strategies can reduce scanning errors and improve image stitching and overlap.For implant restora-tion,when a single crown restoration is supported by implants and a small span upper structure restoration,it is recom-mended to first pre-scan the required dental arch.Then the cavity in the abutment area is excavated.Lastly,scanning the cavity gap after installing the implant scanning rod.When repairing a bone level implant crown,an improved indi-rect scanning method can be used.The scanning process includes three steps:First,the temporary restoration,adjacent teeth,and gingival tissue in the mouth are scanned;second,the entire dental arch is scanned after installing a standard scanning rod on the implant;and third,the temporary restoration outside the mouth is scanned to obtain the three-di-mensional shape of the gingival contour of the implant neck,thereby increasing the stability of soft tissue scanning around the implant and improving scanning restoration.For dental implant fixed bridge repair with missing teeth,the mobility of the mucosa increases the difficulty of scanning,making it difficult for scanners to distinguish scanning rods of the same shape and size,which can easily cause image stacking errors.Higher accuracy of digital implant impres-sions can be achieved by changing the geometric shape of the scanning rods to change the optical curvature radius.The consensus confirms that as the range of scanned dental arches and the number of data concatenations increases,the scanning accuracy decreases accordingly,especially when performing full mouth implant restoration impressions.The difficulty of image stitching processing can easily be increased by the presence of unstable and uneven mucosal mor-phology inside the mouth and the lack of relatively obvious and fixed reference objects,which results in insufficient ac-curacy.When designing restorations of this type,it is advisable to carefully choose digital intraoral scanning methods to obtain model data.It is not recommended to use digital impressions when there are more than five missing teeth.

Objective To explore the value of pelvic CT angiography(CTA)digital three-dimensional reconstruction model(abbreviated as"three-dimensional model")in the diagnosis of female pelvic mass.Methods A total of 98 patients with pelvic mass who were hospitalized and operated in Xi'an People's Hos-pital(Xi'an Fourth Hospital)from January 2021 to April 2023 were selected.All patients underwent B-ultra-sound and CTA examination before operation,and the original data of CTA were collected.The digital three-dimensional model of pelvic mass was established by three-dimensional reconstruction software,and the source of pelvic mass was judged according to the blood supply of pelvic mass.Taking postoperative pathological di-agnosis as the gold standard,the coincidence rate between different preoperative diagnosis methods(B-ultra-sound,CTA examination and three-dimensional model)was compared.The receiver operating characteristic(ROC)curve was plotted to evaluate the efficacy of different preoperative diagnostic methods in judging the ovarian origin of pelvic tumors.Results A total of 130 pelvic masses were included in 98 patients,and the average maximum diameter of the mass was(71.61±3.03)mm,including 83 ovarian masses and 47 non-ovarian masses.Taking postoperative pathological diagnosis as the gold standard,the diagnostic coincidence rate of the preoperative three-dimensional model was 72.31％,which was higher than that of B-ultrasound(58.46％)and CTA(52.31％),and the differences were statistically significant(P＜0.001).The sensitivity,specificity,positive predictive value,negative predictive value,accuracy,Kappa value,and area under the ROC curve were 79.51％,91.49％,94.29％,71.67％,83.85％,0.67 and 0.855,respectively,when the three-dimensional model showed that the blood supply of the mass originated from ovarian artery or uterine artery-ovarian branch.Conclusion The three-dimensional model of pelvic CTA can directly display the blood supply source,characteristics of mass,and the relationship between mass and adjacent organs,which can guide the clinical treatment.It has certain clinical value to judge the ovarian origin of pelvic mass by using ovarian artery and uterine artery-ovarian branch.

Background Animal studies have shown that exposure to organophosphate esters (OPEs) disturbs the composition of gut microbiome in rodents and zebrafish. However, current associated evidence in humans is limited. Considering the importance of gut microbiome in neonatal development, we need to investigate the impact of OPEs exposure on the early development stage of neonatal microbiome. Objective To investigate the associations between umbilical OPEs exposure and the diversity and composition of gut microbiome in newborns. Methods Based on the Shanghai Maternal-Child Pairs Cohort (MCPC), 391 mother-infant pairs with comprehensive follow-up information and bio-samples were enrolled in this study. Concentrations of OPEs in neonatal cord blood were quantified using ultra performance liquid chromatography-tandem mass spectrometry. Meconium samples were collected after delivery and measured through 16S rRNA sequencing on the Illumina Miseq platform. Multiple linear regression models were used to assess the effects of OPEs exposure on the alpha diversity of meconium microbiome. Principal coordinate analysis and permutational multivariate analysis of variance based on unweighted UniFrac distance were used to compare the beta diversity differences between high and low exposure groups of OPEs. Linear discriminant analysis effect size (LEfSe) was utilized to analyze the differential gut microbiome taxa between high and low OPEs exposure groups. The functional pathways involved in the meconium microbiome were predicted based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, and multivariate analysis by linear models (MaAsLin2) were conducted to explore the effects of OPEs exposure on gut microbiome pathways. Results Seven OPEs were detectable in the neonatal cord blood samples, of which four were detected higher than 50% including tributyl phosphate (TBP), tris (2-butoxyethyl) phosphate (TBEP), 2-ethylhexyl diphenyl phosphate (EHDPP), and tris (2-chloro-1 (chloromethyl) ethyl) phosphate (TDCPP), and the median concentrations of these four congeners were as follows: 0.52 μg·L⁻¹ for TBP, 2.41 μg·L⁻¹ for TBEP, 0.13 μg·L⁻¹ for EHDPP, and 2.23 μg·L⁻¹ for TDCPP. A significant association was observed between umbilical TBEP and TDCPP exposure and alpha diversity indices in neonatal meconium microbiome. Beta diversity significantly differed across varied high and low OPEs exposure groups. The results of LEfSe analysis indicated a significant correlation between umbilical OPEs exposure and 27 genera, including Streptococcus, Corynebacterium, Neisseria, Haemophilus, and Parabacteroides. The MaAsLin2 analysis identified associations between OPEs exposure and upregulation of pathways related to linoleic acid metabolism, steroid biosynthesis, Toll and Imd signaling pathway, retinol metabolism, NOD like receptor signaling pathway, and fatty acid biosynthesis . Conclusion Umbilical OPEs exposure is associated increased alpha diversity indices, increased relative abundances of Neisseria, Streptococcus, Parabacteroides, and Corynebacterium in the gut microbiome, as well as predicted metabolic pathway alterations in linoleic acid metabolism, fatty acid biosynthesis, etc. These findings indicate that umbilical OPEs exposure may disrupt meconium microbiome equilibrium.

AIM: To observe the changes of corneal densitometry(CD)in patients with keratoconus after corneal cross-linking(CXL).METHODS: Retrospective study. A total of 32 patients(43 eyes)with keratoconus in Ningxia Eye Hospital from April 2020 to April 2022 were selected. Pentacam analysis system divided the cornea into three layers: anterior 120 μm, middle layer and posterior 60 μm, and divides it into five regions with diameters of 0-2, 2-6, 6-10, 10-12 mm and full diameter according to the diameter, and measures the CD in different ranges. The changes of CD were compared before operation and at 1, 3 and 6 mo after operation.RESULTS: There were differences in uncorrected visual acuity, best corrected visual acuity and intraocular pressure before and 6 mo after operation(all P<0.05), and there was no difference in corneal endothelial cells(P=0.477). CD reached its peak at 1 mo after operation, and decreased at 3 mo and 6 mo after operation, but it was still higher than that before operation. There is a significant positive correlation between CD and Kmax in the anterior layer and the whole layer(r=0.164, P=0.016; r=0.152, P=0.023).CONCLUSION: The values of CD peaked at 1 mo after CXL, then it gradually decreased, tending to become stable at 6 mo postoperatively.