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.

Aim To evaluate the effectiveness and safety of remimazolam tosylate for administering general anesthesia in morbidly obese patients.Methods This clinical trial was conducted at a single center from De-cember 2021 to October 2023.It assessed 108 morbid-ly obese patients(body mass index,BMI≥40)who underwent laparoscopic sleeve gastrectomy.Patients were randomly assigned to either the remimazaolam group(Group R)or the propofol group(Group P)for general anesthesia induction and maintenance.The primary outcome was to compare the incidence of ad-verse events and postoperative recovery characteristics between the two groups.Results During induction pe-riod,the incidence of adverse events was higher in group P,including hypotension(P＜0.01),hypox-emia(P＜0.05),bradycardia(P＜0.01),and in-creased vasopressor requirement(P＜0.05).The time to loss of consciousness and BIS falling to 60 was shor-ter in group P than in group R(P＜0.01).There were no statistically significant differences between the two groups in terms of postoperative quality of recovery(QoR-40 score),24-hour postoperative pain visual an-alogue scale(VAS)scores and morphine consump-tion.In conclusion,remimazolam tosylate,utilized for anesthesia induction in morbidly obese patients,signif-icantly reduced hypotension and hypoxemia compared to propofol,while it could also maintain similar postop-erative recovery quality.Conclusions Remimazolam is effective in reducing the incidence of hypotension and hypoxaemia during the induction period of general anaesthesia in morbidly obese patients and it is compa-rable to propofol in terms of quality of postoperative re-covery.

Regardless of the underlying etiology, renal fibrosis is the final histological outcome of progressive kidney disease. Unilateral ureteral obstruction (UUO) is an ideal and reproducible experimental rodent model of renal fibrosis, which is characterized by tubulointerstitial inflammatory responses, accumulation of extracellular matrix, tubular dilatation and atrophy, and fibrosis. The magnitude of UUO-induced renal fibrosis is experimentally manipulated by the species chosen, animal age, and the severity and duration of the obstruction, while relief of the obstruction allows the animal to recover from fibrosis. The pathogenesis of renal fibrosis is complex and multifactorial and is orchestrated by activation of renin-angiotensin system (RAS), oxidative stress, inflammatory response, transforming growth factor beta 1-Smad pathway, activated myofibroblasts, cell death (apoptosis, autophagy, ferroptosis, and necroptosis), destruction of intracellular organelles, and signaling pathway. The current therapeutic approaches have limited efficacy. Inhibition of RAS and use of antioxidants and antidiabetic drugs, such as inhibitors of sodium-glucose cotransporter 2 and dipeptidyl peptidase-4, have recently gained attention as therapeutic strategies to prevent renal scarring. This literature review highlights the state of the art regarding the molecular mechanisms relevant to the management of renal fibrosis caused by UUO.

Regardless of the underlying etiology, renal fibrosis is the final histological outcome of progressive kidney disease. Unilateral ureteral obstruction (UUO) is an ideal and reproducible experimental rodent model of renal fibrosis, which is characterized by tubulointerstitial inflammatory responses, accumulation of extracellular matrix, tubular dilatation and atrophy, and fibrosis. The magnitude of UUO-induced renal fibrosis is experimentally manipulated by the species chosen, animal age, and the severity and duration of the obstruction, while relief of the obstruction allows the animal to recover from fibrosis. The pathogenesis of renal fibrosis is complex and multifactorial and is orchestrated by activation of renin-angiotensin system (RAS), oxidative stress, inflammatory response, transforming growth factor beta 1-Smad pathway, activated myofibroblasts, cell death (apoptosis, autophagy, ferroptosis, and necroptosis), destruction of intracellular organelles, and signaling pathway. The current therapeutic approaches have limited efficacy. Inhibition of RAS and use of antioxidants and antidiabetic drugs, such as inhibitors of sodium-glucose cotransporter 2 and dipeptidyl peptidase-4, have recently gained attention as therapeutic strategies to prevent renal scarring. This literature review highlights the state of the art regarding the molecular mechanisms relevant to the management of renal fibrosis caused by UUO.

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.

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.

Zuoguiwan， one of the classic formulas of traditional Chinese medicine （TCM）， has the effects of nourishing genuine Yin， supplementing essence， and replenishing marrow， and it is widely used in the clinical treatment of diseases caused by congenital essence deficiency. This article systematically reviewed the ancient books involving congenital deficiency and the modern research reports on the treatment of congenital deficiency with Zuoguiwan. From the formulation principle of Zuoguiwan and the TCM concept of congenital deficiency， this article discussed the theoretical basis of treating congenital deficiency with Zuoguiwan based on the modern research results. Ancient physicians have discovered that the deficiency of parents can lead to abnormal physical development and weakness of the offspring. ZHANG Jingyue divided congenital essence Qi into genuine Yin and primordial Yang and formulated Zuoguiwan based on the principle of Yin-Yang mutual assistance for supplementing congenital genuine Yin. Experimental studies have shown that Zuoguiwan can up-regulate the expression of connexin 43 （Cx43） protein and improve gap junction intercellular communication （GJIC） function to promote osteogenic differentiation and maturation of bone marrow mesenchymal stem cells and ameliorate abnormal bone development caused by congenital essence deficiency. Zuoguiwan can enhance the expression and activity of Reelin to ameliorate abnormal brain development. It can upregulate the expression of Cx43 protein to intervene in the phosphatidylinositol 3 kinase/protein kinase B （PI3K/Akt） pathway， thus repairing the reproductive functions. Zuiguiwan can promote the development and maturation of T cells and activate the Wnt/β-catenin signaling pathway in the thymus to improve immune functions. In addition， it can promote the expression of β-catenin and inhibit the expression of microtubule-associated proteins 1 light chain 3 （LC3） to attenuate skin dysfunctions. Moreover， Zuoguiwan can guide the differentiation of stem cells via the correlation between essence and vitality in TCM. Zuoguiwan has demonstrated significant therapeutic effects on some diseases in the pediatric， andrological， gynecological， geriatric， internal medicine， orthopedic and skin disease departments. On the basis of the results of experiments and clinical applications， this paper analyzes the specific connotation of congenital deficiency proposes that congenital deficiency should be subdivided into the four aspects of essence， Qi， Yin， and Yang. Congenital essence deficiency as the deficiency of genuine Yin can lead to deficiency of kidney essence in the offspring and dysfunction of kidney storing essence. Pharmacological studies have discovered that Cuscutae Semen， Lycii Fructus， and Achyranthis Bidentatae Radix in Zuoguiwan contain active components such as quercetin and kaempferol， which act on the targets such as recombinant prostaglandin endoperoxide synthase 2 （PTGS2）， interleukin-6 （IL-6）， mitogen-activated protein kinases （MAPK14）， tumor necrosis factor （TNF）， TP53， Vascular permeability factor A （VEGFA）， and albumin （ALB） to play a role in reproductive system development and hormone responses. Zuoguiwan has unique advantages over Liuwei Dihuangwan in nourishing congenital genuine Yin in the kidney and can achieve better therapeutic effects on the syndromes and diseases caused by congenital essence deficiency. This review is expected to enrich the knowledge about the efficacy and clinical application of Zuoguiwan， provide new perspectives and methods for the prevention and treatment of congenital deficiency and congenital essence deficiency， and give insights into the application of Zuoguiwan in modern healthcare， especially in the nurturing of offspring.

Objective To analyze the clinical significance of elevated CHMP2B expression in pancreatic cancer,and to explore the oncogenic mechanism of CHMP2B in pancreatic cancer.Methods A transcriptomic analysis conducted on the GEPIA website revealed CHMP2B expression in over 20 types of malignant tumors.The relation-ship between CHMP2B expression and disease-free survival and overall survival of pancreatic cancer was analyzed.The correlation between CHMP2B,MAP1LC3B and P62 expression was analyzed by TCGA database.The overex-pression plasmid of CHMP2B factor was transfected with Lipofectamine 2000 in MIA PaCa-2 cells,and the overex-pression of CHMP2B factor in MIA PaCa-2 was verified by Western blot.The effect of overexpression of CHMP2B on tumorigenic ability of MIA PaCa-2 cells was verified by clonal formation experiment.The effect of overexpression of CHMP2B on proliferation,migration and invasion of MIA PaCa-2 cells was verified by CCK-8 and Transwell ex-periments.The localization of CHMP2B factor was detected by immunofluorescence staining.The correlation be-tween CHMP2B,MAP1LC3B and P62 expression was analyzed by TCGA database.The expression of MAP1LC3B and P62 in CHMP2B overexpression was verified by qPCR.Results The expression of CHMP2B factor increased in pancreatic cancer(P＜0.01).Overexpression of CHMP2B factor was associated with overall survival and dis-ease-free survival(P＜0.01).The high expression of CHMP2B promoted the tumorigenesis of MIA PaCa-2 cells in pancreatic cancer,and also promoted the proliferation,migration and invasion of pancreatic cancer.Immunofluo-rescence staining showed that CHMP2B was distributed in both nucleus and cytoplasm and correlated with vesicle secretion.Moreover,CHMP2B was positively correlated with the expression of autophagy marker MAP1LC3B while inhibiting the expression of P62.Conclusion The high expression of CHMP2B in clinical pancreatic cancer tissue samples is consistent with the high expression in vitro pancreatic cancer cell lines.CHMP2B is localized in both nu-cleus and cytoplasm,involving in vesicle transport and lysosomal autophagy.In the pancreatic cancer cell line MIA PaCa-2,CHMP2B is positively correlated with the expression of autophagy marker MAP1LC3B and inhibits the ex-pression of P62.It is concluded that CHMP2B is related to autophagy in promoting the malignant degree of pancre-atic cancer.CHMP2B meets the high nutritional metabolic requirements of pancreatic cancer cells by regulating ve-sicular transport and autophagy,thus promoting the malignant biological behavior of pancreatic cancer.

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.