BACKGROUND:Streptococcus mutans is an important pathogen of dental caries,and timely detection of its levels is of great significance for early detection and treatment of dental caries. OBJECTIVE:To evaluate the effect of loop-mediated isothermal amplification(FTA-LAMP)direct extraction of Streptococcus mutans DNA. METHODS:(1)Bacterial suspensions containing ATCC standard strains(Streptococcus mutans)were prepared and inoculated into the brain-heart leachate medium.After mixed thoroughly,the mixture was then diluted in a 10-fold gradient into seven concentrations(4.2×107,4.2×106,4.2×105,4.2×104,4.2×103,4.2×102,4.2×10 CFU/mL),two parallel controls were made for each dilution level,and sterile water was used as a blank control.(2)The DNA of Streptococcus mutans was extracted using FTA Elute card,boiling method,kit extraction and lysate extraction methods separately and then amplified using LAMP technology was amplified.A specificity test was also performed to compare the differences between the four DNA extraction methods.RESULTS AND CONCLUSION:The DNA extracted by all four methods met the requirements for LAMP amplification.Specificity test results showed that only Streptococcus mutans could specifically amplify the target gene.The detection limit value of the DNA concentration was 4.2×103 CFU/mL for the lysate method,4.2×104 CFU/mL for the FTA Elute card extraction method,4.2×106 CFU/mL for the kit extraction method,and 4.2×107 CFU/mL for the boiling method.In the other aspects of the four extraction methods,the kit extraction method had the highest experimental cost,number of steps and time;the other three methods had the same number of steps,with the FTA Elute card method requiring the least amount of instruments,the boiling method having the lowest single cost,and the lysate extraction method taking the least amount of time.Only a small amount of bacteria were needed for successful extraction using both the FTA Elute card and lysate extraction methods.Compared with the FTA Elute card method,the lysate extraction method was superior in terms of time,but it had a high single cost and required more equipment.To conclude,the FTA-LAMP technology established in this study has the advantages of ease of operation,high specificity,high sensitivity,and visualization,which is expected to be a new way for efficient extraction and detection of Streptococcus mutans.

As a mercury-containing elixir， Hongshengdan has been known as a sacred medicine for surgery by ancient medical practitioners because of its precise curative effects. It originated from Yizong Shuoyue in the Qing dynasty， Qing dynasty and modern medical practitioners have adapted and modified its formula for clinical application. Employing bibliometric methods， the authors systematically organized relevant ancient literature of the Qing dynasty and modern literature， and analyzed the composition and dosage， preparation method， and clinical application. Among the 25 ancient books concerning Hongshengdan， a total of 12 medicinal formulas， 15 refining methods and 9 clinical applications were obtained. Research confirms that Hongshengdan consisted of mercury， saltpeter， alum， soap alum， cinnabar and realgar. Using measurement conversion standards of Qing dynasty， the modern single-batch formulation comprised 37.30 g of mercury， 149.20 g of saltpeter， 37.30 g of alum， 22.38 g of soap alum， 18.65 g of cinnabar， and 18.65 g of realgar. In modern refining of Hongshengdan， most medical practitioners take the core medicines， with dosages approximately 30 g of mercury， 30 g of saltpeter， and 30 g of alum. Refining method involves pretreatment stewing the materials during preparation， and alum， soap alum， and saltpeter are first ground together， then combined with mercury， cinnabar， and realgar for grinding until mercury and other drugs grind to the degree of no star points. The mixture is then placed in a pot or vessel by cold-forming method. After covering， the opening is sealed using either raw gypsum salt mud or honey-dipped cotton paper strips. Sand is packed around the vessel and then pressurized. During the calcination process， begin with a low flame（30 min）， then increase to a medium flame（30 min）， followed by a high flame（30 min）， after removing fire toxins， collect the final product. Hongshengdan has the efficacy of lifting the poison， removing the corrosion， producing muscle and dispersing， and is often used in the treatment of surgical sore and carbuncle type of diseases. Modern research indicates that Hongshengdan is commonly used to treat skin system diseases such as ulcers and herpes. The aforementioned findings provide a reference basis for the subsequent refining method and clinical application of Hongshengdan.

Objective:To summarize and analyze the acupoint selection law of acupuncture and moxibustion for the treatment of schizophrenia using data mining techniques and network visualization technology.Methods:The literature related to the treatment of schizophrenia through acupuncture and moxibustion was retrieved from CNKI, Wanfang, VIP, SinoMed, Chinese Medical Journal Full-text Database, PubMed and Web of Science from the establishment of the databases to December 31, 2022. A database was established using Excel 2013 software and frequency analysis of acupoints was conducted; the Apriori algorithm was used to analyze the association law of acupoints; SPSS Statistics 25.0 and SPSS Modeler 18.0 software were used for acupoint clustering analysis to explore the acupoint selection law.Results:A total of 133 articles were included, involving 133 acupuncture prescriptions and 158 acupoints, with a total frequency of 1131 times. The most frequently used acupoints were Baihui (GV20) (71 times), Zusanli (ST36) (51 times), Neiguan (PC6) (45 times). Commonly used meridians were Governor Vessel, and stomach meridian, etc. According to the analysis of association rules, 108 association rules of acupoints were obtained, and common compatibility of acupoints was Baihui (GV20)-Yintang (GV24)-Zusanli (ST36), Baihui (GV20)-Shenting (GV24), Baihui (GV20)-Yintang (GV24), Daling (PC7)-Laogong (PC8), etc. Four major clusters were obtained through clustering analysis of commonly used acupoints.Conclusion:Modern acupuncture and moxibustion is mainly used to regulate yin and yang and balance qi and blood in the treatment of schizophrenia. The acupoint selection is based on Baihui-Yintang-Neiguan-Hegu acupoint prescription, and other acupoints are added according to concurrent syndromes, which can not only give full play to the synergistic effect of acupoints, but also promote the recovery of motor and sensory functions.

Acute cerebral infarction(ACI)has the characteristics of onset nasty and high mortality,and thus the rapid determination of the occurrence and development of ACI plays a key role in the diagnosis,treatment and prognosis of ACI patients.It has shown that the serum level of human haptoglobin(Hp)is related to ACI.In this study,surface enhanced Raman scattering(SERS)combined with immune recognition was applied to establish a quantitative analysis method for serum Hp.Firstly,the SERS substrate of silver nanoparticles was prepared on silicon wafer,and 4-mercaptobenzoic Acid(MBA)was used as a Raman probe by forming Ag—S bond and connecting it on the surface of nanoparticles.The carboxyl group of MBA was linked to amino group of self-made high-affinity antibody through forming CO—NH structure thus forming a SERS self-assembled chip of Hp(Ag/MBA/anti-Hp).Hp in serum could be specifically captured by antibodies on SERS substrate,which caused the shift of SERS characteristic peak of MBA.The results showed that there was a good linear relationship between the logarithm of Hp concentration and the SERS characteristic peak shift of MBA.The detection range was 1-1000 ng/mL(R2=0.988).The Hp concentrations in serum of 90 ACI patients were determined by this method,and the results were consistent with those of ELISA method,which proved the practicability and accuracy of this method.This method was highly specific,simple and convenient,which could realize the specific recognition and quantitative analysis of serum Hp,so as to be an effective means for clinical detection of serum Hp,thus providing a reference for the treatment and prognosis of ACI.

OBJECTIVE To evaluate the efficacy， safety and cost-effectiveness of vericiguat in the treatment of heart failure （HF） by rapid health technology evaluation method， and provide reference for the selection and decision-making of clinical treatment plans. METHODS Chinese and foreign databases such as CNKI， PubMed and related health technology evaluation websites were searched by computer. Relevant researchers independently screened literature， extracted data， and comprehensively analyzed the results of the included literature based on literature quality evaluation. RESULTS A total of 17 pieces of literature were included， involving 12 systematic reviews/meta-analyses and 5 pharmacoeconomic studies. The effectiveness analysis showed： for HF patients， compared with placebo， vericiguat （10 mg/d） significantly improved the EuroQol five dimensions questionnaire （EQ-5D） index and decreased the rate of hospitalization due to HF （P＜0.05）. For heart failure with reduced ejection fraction （HFrEF） patients， vericiguat reduced the incidence of hospitalization due to HF compared with sodium-glucose cotransporter 2 inhibitor （SGLT2i）（P＜0.05）； compared with angiotensin-converting enzyme inhibitor， vericiguat significantly reduced the occurrence risk of composite endpoints of cardiovascular death or hospitalization due to HF（P＜0.05）. For HFrEF patients with chronic kidney disease， vericiguat had a tendency to reduce the occurrence risk of composite endpoints of cardiovascular death or hospitalization due to HF compared with neurohormone inhibitors. Safety analysis showed： vericiguat did not increase drug-related adverse reactions compared to placebo （P＞0.05）. Economic analysis showed： domestic studies indicated that vericiguat had a higher incremental cost-effectiveness ratio. CONCLUSIONS Vericiguat has good safety and efficacy in the treatment of HF but does not possess an economic advantage in the Chinese population.

Purpose::Under-foot impact loadings can cause serious lower limb injuries in many activities, such as automobile collisions and underbody explosions to military vehicles. The present study aims to compare the biomechanical responses of the mainstream vehicle occupant dummies with the human body lower limb model and analyze their robustness and applicability for assessing lower limb injury risk in underfoot impact loading environments.Methods::The Hybrid III model, the test device for human occupant restraint (THOR) model, and a hybrid human body model with the human active lower limb model were adopted for under-foot impact analysis regarding different impact velocities and initial lower limb postures.Results::The results show that the 2 dummy models have larger peak tibial axial force and higher sensitivity to the impact velocities and initial postures than the human lower limb model. In particular, the Hybrid III dummy model presented extremely larger peak tibial axial forces than the human lower limb model. In the case of minimal difference in tibial axial force, Hybrid III's tibial axial force (7.5 KN) is still 312.5% that of human active lower limb's (2.4 KN). Even with closer peak tibial axial force values, the biomechanical response curve shapes of the THOR model show significant differences from the human lower limb model.Conclusion::Based on the present results, the Hybrid III dummy cannot be used to evaluate the lower limb injury risk in under-foot loading environments. In contrast, potential improvement in ankle biofidelity and related soft tissues of the THOR dummy can be implemented in the future for better applicability.

Polysaccharides are the main active ingredients in traditional Chinese medicine and ideal raw materials for new drug development with diverse structures,rich activity and low toxicity.Research has shown that traditional Chinese medicine polysaccharides(TCMP)exhibit outstanding associated activities in anti-tumor,immunomodulatory,anti-fibrotic,and Alzheimer's disease prevention,with clear mechanisms and targeting properties.In this review,we discuss research progress of the mechanisms and discovery of target molecules of TCMP in various areas,including anticancer,immunomodulation,gut microbiota regulation,anti-fibrosis,and Alzheimer's disease prevention.By analyzing relevant literature,it has been observed that TCMP exhibits various complex mechanisms of action,highlighting significant targeting properties in immunomodulation,anticancer effects and disrupts angiogenesis activities.At the same time,this review summarizes and analyzes the current shortcomings and challenges in the research on TCMP and proposes future directions,aiming to provide new ideas and strategies for understanding the basis of active substances in traditional Chinese medicine and the development of original new drugs based on polysaccharides.

Objective This study aims to investigate the structure and anti-angiogenic activity of homogeneous polysaccharides in Honeysuckle flowers,constructing the theoretical basis for its widespread application.Methods Homogeneous Honeysuckle polysaccharide LF-02-2 was obtained through water extraction,alcohol precipitation,anion exchange chromatography,and gel permeation chromatography.The structure of LF-02-2 was deduced through molecular weight determination,monosaccharide composition analysis,sugar residue linkage analysis,partial acid hydrolysis,and sugar aldonic acid reduction combined with NMR data.Additionally,in vitro tube formation experiments using human microvascular endothelial cells(HMEC-1)were conducted to assess its anti-angiogenic activity.Results Structural analysis of LF-02-2 revealed a homogeneous polysaccharide with a weight-average molecular weight of 74.1 kDa.The monosaccharide composition included rhamnose(Rha),galactose(Gal),galacturonic acid(GalA),and arabinose(Ara)in molar ratios of 10.43:14.94:6.66:67.97.The main chain consisted of 1,4-linked α-Galp A,1,2-linked α-Rhap,and 1,2,4-linked α-Rhap.Branches were connected to O-4 of 1,2,4-linked α-Rhap and included β-Galp with terminal connections and 1,4,6-linked β-Galp and α-Araf with terminal and 1,5-linked connections.Tube formation experiments demonstrated that LF-02-2 significantly inhibited tube formation in human microvascular endothelial cells(HMEC).Conclusion LF-02-2,an RG-I pectic polysaccharide from Honeysuckle flowers,exhibited significant anti-angiogenic activity,suggesting its potential for development as an anti-angiogenic drug.

Objective This study aims to elucidate the structure-activity domain of LBP1C-2 targeting Kvβ.2 through an exploration of the structure-activity relationship.This study may also provide the scientific basis for the development of drug candidate with anti-early-onset dementia activity.Methods After partial acid hydrolysis,various structural fragments were obtained and subjected to monosaccharide composition and molecular weight analysis.Potential target proteins were selected using a protein chip,followed by validation of the targeting specificity of each structural fragment using surface plasmon resonance(SPR)technology.Results Through high-throughput screening using the HuProtTM human protein array,potential target protein Kvβ.2 was identified for LBP1C-2.SPR experiments revealed a strong binding affinity between LBP1C-2 and Kvβ.2 protein,with a binding constant(KD)of 1.9×10-7 M.The various structural fragments of LBP1C-2 exhibited different binding strengths with the target protein Kvβ.2.Among them,the segment LBP1C-2-1I(18.1 k Da)with a molar ratio of rhamose to galecturonic acid of 1:1 showed a binding strength to Kvβ.2 similar to that of the polysaccharide LBP1C-2,with a KD of approximately 3.3×10-7 M.Structural analysis indicates that the structure of LBP1C-2-1I contains 1,2-linked Rha and 1,4-linked GalA which are alternatively linked.The acid-hydrolyzed extracellular portion corresponding to this segment,LBP1C-2-1O may also bind to Kvβ.2.However,compared to other segments,it demonstrated a higher tendency to dissociate from the protein.Knockdown of the KCNAB2 gene(Kvβ.2)in BV2 cells inhibited the uptake of Aβ in BV2 cells,suggesting that protein Kvβ.2 may be a functional protein in the development of Alzheimer's disease.Conclusion LBP1C-2-1I has been identified as the primary active domain through which LBP1C-2 targets Kvβ.2.This suggests that the active domain of LBP1C-2 predominantly resides on the main chain rather than the side chain.This study provides crucial insights for a deeper understanding of the anti-early-onset dementia activity of LBP1C-2 and lays an experimental foundation for the design and development of targeted drugs for anti-early-onset dementia based on Lycium barbarum polysaccharides.

Objective To elucidate the structural basis of polysaccharides with hypoglycemic activity in mulberry leaves,and provide a theoretical basis for the development and utilization of mulberry leaves.Methods The crude polysaccharides from mulberry leaves were extracted by hot water.The components of SY02,SY02-3,SY02-3A and SY02-3B were obtained by ion-exchange chromatography and gel permeation chromatography.We determined the physicochemical properties of homogeneous polysaccharide.The structure of mulberry leaf polysaccharide was identified by monosaccharide composition analysis,sugar residue linkage analysis and nuclear magnetic analysis.Palimitate(PA)induced INS-1 cell apoptosis model was used to determine the protective effect of mulberry leaf polysaccharide on INS-1E cell apoptosis,and Caspase3/7 activity kit was used to detect mulberry leaf polysaccharide on the key protein of INS-1E cell apoptosis Cleaved by Western blot method effect of Caspase 3 expression.Results The crude polysaccharide SY was extracted from mulberry leaves by hot water,which was separated by ion exchange resin column and purified by gel column to obtain SY02-3.After purification,we obtained homogeneous polysaccharide SY02-3A and proteoglycan SY02-3B,with molecular weights of 3.7×104 Da and 1.03×104 Da.The results of monosaccharide composition analysis showed that SY02-3A contained rhamnose,galacturonic acid,galactose,xylose and arabinose.The molar ratio of SY02-3A was 23.97:33.85:11.73:5.04:25.41,and the yield was 0.32%.SY02-3B is composed of rhamnose,galacturonic acid,glucose,galactose and arabinose.The molar ratio of SY02-3B is 19.83:10.34:45.42:9.01:2.23:13.17 with a yield of 2.00%.SY02-3B contains 14 amino acids,among which the content of aspartate,glutamic acid,alanine and glycine is relatively high.We also found that SY02-3 protects palmitic acid-induced INS-1E cell apoptosis,which may be associated with reduced Caspase 3 activity and down-regulated cleaved Caspase 3 protein expression.Conclusion Mulberry leaf polysaccharide SY02-3,which contains acid peptidoglycan(SY02-3A)and proteoglycan(SY02-3B)in mulberry leaves,has the effect of inhibiting PA-induced apoptosis of INS-1E cells.