ObjectiveThis paper aims to investigate and evaluate the staged efficacy and safety of the representative empirical prescription of the “Zhu Ke Jiao” theory， Qijia Rougan prescription， combined with entecavir in the treatment of hepatic fibrosis in chronic hepatitis B. MethodsA multicenter randomized controlled clinical study was conducted， and 101 patients diagnosed with chronic hepatitis B-related hepatic fibrosis （CHB-HF） who met the diagnosis and inclusion criteria were randomly assigned to an observation group （Qijia Rougan prescription + entecavir） and a control group （entecavir）. The treatment duration was 24 weeks. Liver stiffness measurement （LSM）， fibrosis-4 index （FIB-4）， portal vein diameter， hepatitis B serology， biochemical indicators， hepatic fibrosis markers in serum ［hyaluronic acid （HA）， laminin （LN）， procollagen Ⅲ peptide （PⅢP）， and type Ⅳ collagen （Ⅳ-C）］， and traditional Chinese medicine syndrome scores were used as efficacy evaluation indicators. Efficacy assessments and explorations of different staged subgroups of Qijia Rougan prescription were conducted according to LSM values based on the Metavir pathological staging standard. ResultsA total of 98 cases were included for statistical analysis， with 49 cases in the observation group and 49 in the control group. The general data of the patients in both groups were comparable. Compared with the same group before treatment， the observation group showed a significant reduction in LSM and FIB-4 （P<0.01）， as well as notable improvements in LN， Ⅳ-C， and various TCM syndrome scores （P<0.05， P<0.01）. When compared to the control group after treatment， the observation group demonstrated significant improvements in LSM， FIB-4， and various TCM syndrome score indicators （P<0.05， P<0.01）， indicating that the observation group performed better than the control group. Subgroup analysis of the regression of hepatic fibrosis stages showed that compared to the same group before treatment， the observation group had better improvement in regression of stages F2 and F3 （P<0.05）. When compared to the control group after treatment， the observation group exhibited superior improvement in regression of stage F3 （P<0.05）. No adverse events occurred in either group during the treatment period. ConclusionCompared with entecavir alone， the combination of Qijia Rougan prescription and entecavir significantly improves the degree of hepatic fibrosis and clinical TCM symptoms in patients. The optimal intervention period is primarily during stage F3， which is a potential “interception” point of the “Zhu Ke Jiao” theory.

OBJECTIVE To focus on the classic NOD-like receptor protein 3 （NLRP3）/Caspase-1/gasdermin D （GSDMD） pyroptosis pathway and explore the mechanism by which Huatan qushi huoxue formula （HQHF） inhibits macrophage pyroptosis to ameliorate metabolic dysfunction-associated steatohepatitis （MASH）. METHODS RAW264.7 cells were divided into 5 groups： Control group （10% blank serum）， Model group [10% blank serum+5 μg/mL lipopolysaccharide （LPS）]， HQHF-L group （2.5% drug-containing serum+7.5% blank serum+5 μg/mL LPS）， HQHF-M group （5% drug-containing serum+5% blank serum+5 μg/mL LPS）， and HQHF-H group （10% drug-containing serum+5 μg/mL LPS）. After 24 h of routine culture post-administration， cells and supernatants were collected for assays. Cell morphology was observed via scanning electron microscopy and phase-contrast microscopy； localization and expression of gasdermin D-N （GSDMD-N） were observed by immunofluorescence. Interleukin-1β （IL-1β） and IL-18 contents in supernatants were detected by ELISA； mRNA and protein expressions of NLRP3， Caspase-1， and GSDMD were measured using real-time PCR and Western blot. RESULTS Compared with the Control group， the Model group showed typical pyroptotic morphology （cell membrane bulging and pore formation）， increased aggregation and fluorescence intensity of GSDMD-N on the cell membrane （ P ＜0.05）， significantly increased the contents of IL-1β and IL-18 in cell supernatants （ P ＜0.05）， and significantly up-regulated mRNA and protein expressions of NLRP3， Caspase-1， and GSDMD in cells （ P ＜0.05）. Compared with the Model group， the HQHF-L， HQHF-M and HQHF-H groups showed improved pyroptotic morphology， reduced membrane localization and significantly weakened fluorescence intensity of GSDMD-N （ P ＜0.05）， significantly decreased the contents of IL-1β and IL-18 in cell supernatants （ P ＜0.05）， and significantly down-regulated mRNA and protein expressions of NLRP3， Caspase-1， and GSDMD in cells （ P ＜0.05）. CONCLUSIONS HQHF inhibits LPS-induced macrophage pyroptosis， and its mechanism of improving MASH may be associated with the suppression of the activation of the classical NLRP3/Caspase-1/GSDMD pyroptosis pathway.

Oral squamous cell carcinoma (OSCC) is a common head and neck malignancy. Approximately 50% to 60% of patients with OSCC are diagnosed at a locally advanced stage (clinical staging III-IVa). Even with comprehensive and sequential treatment primarily based on surgery, the 5-year overall survival rate remains below 50%, and patients often suffer from postoperative functional impairments such as difficulties with speaking and swallowing. Programmed death receptor-1 (PD-1) inhibitors are increasingly used in the neoadjuvant treatment of locally advanced OSCC and have shown encouraging efficacy. However, clinical practice still faces key challenges, including the definition of indications, optimization of combination regimens, and standards for efficacy evaluation. Based on the latest research advances worldwide and the clinical experience of the expert group, this expert consensus systematically evaluates the application of PD-1 inhibitors in the neoadjuvant treatment of locally advanced OSCC, covering combination strategies, treatment cycles and surgical timing, efficacy assessment, use of biomarkers, management of special populations and immune related adverse events, principles for immunotherapy rechallenge, and function preservation strategies. After multiple rounds of panel discussion and through anonymous voting using the Delphi method, the following consensus statements have been formulated: 1) Neoadjuvant therapy with PD-1 inhibitors can be used preoperatively in patients with locally advanced OSCC. The preferred regimen is a PD-1 inhibitor combined with platinum based chemotherapy, administered for 2-3 cycles. 2) During the efficacy evaluation of neoadjuvant therapy, radiographic assessment should follow the dual criteria of Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 and immune RECIST (iRECIST). After surgery, systematic pathological evaluation of both the primary lesion and regional lymph nodes is required. For combination chemotherapy regimens, PD-L1 expression and combined positive score need not be used as mandatory inclusion or exclusion criteria. 3) For special populations such as the elderly (≥ 70 years), individuals with stable HIV viral load, and carriers of chronic HBV/HCV, PD-1 inhibitors may be used cautiously under the guidance of a multidisciplinary team (MDT), with close monitoring for adverse events. 4) For patients with a poor response to neoadjuvant therapy, continuation of the original treatment regimen is not recommended; the subsequent treatment plan should be adjusted promptly after MDT assessment. Organ transplant recipients and patients with active autoimmune diseases are not recommended to receive neoadjuvant PD-1 inhibitor therapy due to the high risk of immune related activation. Rechallenge is generally not advised for patients who have experienced high risk immune related adverse events such as immune mediated myocarditis, neurotoxicity, or pneumonitis. 5) For patients with a good pathological response, individualized de escalation surgery and function preservation strategies can be explored. This consensus aims to promote the standardized, safe, and precise application of neoadjuvant PD-1 inhibitor strategies in the management of locally advanced OSCC patients.

BACKGROUND:Distinct risk factors for new adjacent vertebral fractures following percutaneous vertebroplasty in the elderly patients with osteoporotic vertebral compression fractures and underlying diseases may be different,yet there is a scarcity of pertinent research on this topic.OBJECTIVE:To investigate the high-risk factors for refracture in patients with osteoporotic vertebral compression fractures following percutaneous vertebroplasty and to delve deeper into the correlation between these risk factors and the recurrence of fractures in the operated vertebra as well as its adjacent vertebrae post-percutaneous vertebroplasty.METHODS:According to the inclusion criteria,412 patients with osteoporotic vertebral compression fractures who underwent percutaneous vertebroplasty at the Sixth Affiliated Hospital of Xinjiang Medical University between January 2018 and December 2023 were retrospectively screened.According to whether re-fracture occurred during the follow-up period,the patients were divided into a re-fracture group(n=75)and a non-re-fracture group(n=337).The following variables of the two groups of patients were reviewed:age,gender,body mass index,cement leakage,bone density T value,cement dosage,occupation,smoking,drinking,adverse reactions,medical insurance,hypertension,type 2 diabetes,chronic obstructive pulmonary disease,cerebral hemorrhage,coronary heart disease,psychological trauma,psychiatric treatment,osteoarthritis,gout,scoliosis,spinal surgery,chronic kidney,anti-osteoporosis treatment,previous fracture history(fracture caused by osteoporosis)and fracture injury plane.The above factors were analyzed using univariate analysis.The factors with P＜0.05 were subjected to binary logistic analysis to explore the related factors of postoperative vertebral recompression fracture.RESULTS AND CONCLUSION:(1)Univariate analysis showed that after percutaneous vertebroplasty,vertebral refracture was associated with age,smoking history of＞10 years,cement leakage,hypertension,type 2 diabetes,scoliosis,spinal surgery,and chronic kidney disease,with statistically significant differences(P＜0.05).(2)By binary Logistic regression,bone cement leakage condition(OR=2.547,95％CI:1.283-5.048,P＜0.05),smoking history＞10 years(OR=2.336,95％CI:1.157-4.701,P＜0.05),a history of hypertension(OR=4.657,95％CI:2.137-10.242,P＜0.05),a history of type 2 diabetes mellitus(OR=8.956,95％CI:3.941-21.301,P＜0.05),and a medical history of scoliosis(OR=3.754,95％CI:1.755-8.619,P＜0.05),medical history of spinal surgery(OR=2.700,95％CI:1.058-6.725,P＜0.05),and a history of chronic kidney disease(OR=2.812,95％CI:1.078-7.739,P＜0.05)were risk factors for refracture of the operated vertebral body and adjacent vertebrae.(3)The results showed that cement leakage,smoking for＞10 years,hypertension,type 2 diabetes mellitus,scoliosis,spinal surgery,and chronic kidney were risk factors for recurrent fractures of the operated vertebrae and adjacent vertebrae in patients with osteoporotic vertebral compression fractures.Surgeons should conduct a comprehensive assessment of patients before percutaneous vertebroplasty in order to more accurately predict the possibility of re-fracture and provide a basis for the formulation of personalized treatment plans to reduce the risk of future recurrent fractures.

BACKGROUND:Distinct risk factors for new adjacent vertebral fractures following percutaneous vertebroplasty in the elderly patients with osteoporotic vertebral compression fractures and underlying diseases may be different,yet there is a scarcity of pertinent research on this topic.OBJECTIVE:To investigate the high-risk factors for refracture in patients with osteoporotic vertebral compression fractures following percutaneous vertebroplasty and to delve deeper into the correlation between these risk factors and the recurrence of fractures in the operated vertebra as well as its adjacent vertebrae post-percutaneous vertebroplasty.METHODS:According to the inclusion criteria,412 patients with osteoporotic vertebral compression fractures who underwent percutaneous vertebroplasty at the Sixth Affiliated Hospital of Xinjiang Medical University between January 2018 and December 2023 were retrospectively screened.According to whether re-fracture occurred during the follow-up period,the patients were divided into a re-fracture group(n=75)and a non-re-fracture group(n=337).The following variables of the two groups of patients were reviewed:age,gender,body mass index,cement leakage,bone density T value,cement dosage,occupation,smoking,drinking,adverse reactions,medical insurance,hypertension,type 2 diabetes,chronic obstructive pulmonary disease,cerebral hemorrhage,coronary heart disease,psychological trauma,psychiatric treatment,osteoarthritis,gout,scoliosis,spinal surgery,chronic kidney,anti-osteoporosis treatment,previous fracture history(fracture caused by osteoporosis)and fracture injury plane.The above factors were analyzed using univariate analysis.The factors with P＜0.05 were subjected to binary logistic analysis to explore the related factors of postoperative vertebral recompression fracture.RESULTS AND CONCLUSION:(1)Univariate analysis showed that after percutaneous vertebroplasty,vertebral refracture was associated with age,smoking history of＞10 years,cement leakage,hypertension,type 2 diabetes,scoliosis,spinal surgery,and chronic kidney disease,with statistically significant differences(P＜0.05).(2)By binary Logistic regression,bone cement leakage condition(OR=2.547,95％CI:1.283-5.048,P＜0.05),smoking history＞10 years(OR=2.336,95％CI:1.157-4.701,P＜0.05),a history of hypertension(OR=4.657,95％CI:2.137-10.242,P＜0.05),a history of type 2 diabetes mellitus(OR=8.956,95％CI:3.941-21.301,P＜0.05),and a medical history of scoliosis(OR=3.754,95％CI:1.755-8.619,P＜0.05),medical history of spinal surgery(OR=2.700,95％CI:1.058-6.725,P＜0.05),and a history of chronic kidney disease(OR=2.812,95％CI:1.078-7.739,P＜0.05)were risk factors for refracture of the operated vertebral body and adjacent vertebrae.(3)The results showed that cement leakage,smoking for＞10 years,hypertension,type 2 diabetes mellitus,scoliosis,spinal surgery,and chronic kidney were risk factors for recurrent fractures of the operated vertebrae and adjacent vertebrae in patients with osteoporotic vertebral compression fractures.Surgeons should conduct a comprehensive assessment of patients before percutaneous vertebroplasty in order to more accurately predict the possibility of re-fracture and provide a basis for the formulation of personalized treatment plans to reduce the risk of future recurrent fractures.

Vaccine immunization is the most effective and cost-efficient method for infectious disease prevention and control.Since the outbreak of novel coronavirus pneumonia(caused by the novel coronavirus,COVID-19)at the end of 2019,third generation mRNA nucleic acid vaccines has been applied to stop viral spread.mRNA vaccines,rather than relying on the immune activation mode of traditional vaccines,are an innovative breakthrough using the body's own cells to produce antigens,thereby activating double specific immunity,forming immune memory,and providing more lasting specific immunity.Com-pared with the traditional first-generation(inactivated)and second-generation(genetically engineered)vaccines,mRNA vac-cines,because of the advantages provided by this platform,play important roles in the prevention and control of major sudden infectious diseases.Consequently,mRNA vaccines were the world's first COVID-19 vaccines to be applied clinically,thus ser-ving as a barometer in the field of vaccine research and development.Herein,the molecular design and presentation of mRNA vaccines,and the molecular mechanisms of their activation of the immune response are reviewed,to provide a theoretical basis for future application of novel mRNA vaccines in the prevention and control of animal infectious diseases.

Objective To analyze the research status,hot directions and frontier trends of traditional Chinese medicine in the prevention and treatment of diseases by regulating Notch signaling pathway based on bibliometrics.Methods Based on Citespace and Vosviewer,the literature on the regulation of Notch signaling pathway by traditional Chinese medicine in CNKI and WoSCC was visually analyzed.Results 362 and 85 related literatures were published in Chinese and English respectively until January 2024.Since 2013,the number of literatures published in this field has shown a fluctuating increasing trend.China is the country with the most publications;Hunan University of Chinese Medicine were the institutions with the most publications in the Chinese database,and Beijing University of Chinese Medicine was the institution with the most publications in the English literature database.Combined with the research direction of each research team and keyword clustering and burst analysis,the research hotspots of traditional Chinese medicine regulating Notch signaling pathway are focused on cerebral ischemia,myelosuppression and hepatic fibrosis.Diseases such as Asthma,colorectal cancer have become emerging research directions in recent years.Electroacupuncture therapy to promote stem cell proliferation and treat neurological diseases is one of the frontier research trends in this field.Conclusion Recent years have seen a rapid development of traditional Chinese medicine's disease prevention and treatment that targets Notch signaling pathway.Various expert teams have obtained rich research results,and the research hotspots show a diversified trend.In-depth exploration of this can provide strong evidence for the molecular mechanism of traditional Chinese medicine in the treatment of various diseases.

The SVA and different serotypes of VSV(VSNJV and VSIV)are susceptible to infect pigs and cause blister injuries to the lips and hoof of pigs.The clinical symptoms of diseases caused by these viruses are very similar,which is easy to cause misdiagnosis.Therefore,a multiplex nano-PCR method was developed for the simultaneous defection of VSV,VSNJV and VSIV.In this stud-y,three pairs of specific primers were designed according to the SVA-P gene,VSNJV-N gene and VSIV-N gene.The optimal annealing temperature and optimal primer concentration were tested,and the reaction system and conditions were optimized.We have developed a novel,rapid and sensitive multiple nano-PCR detection method for simultaneous detection of SVA,VSNJV and VSIV,which was developed by using nano-metal materials.The specific test results showed that the method could specifically amplify the target genes of SVA,VSNJV and VSIV,with no cross-reactivity to PRV,ASFV,PCV2 and PHEV.The sensitivity test results showed that the minimum nucleic acid detection of the method was 10 copies/μL,which sensitivity was great.In addition,the optimal primers showed good reactivity and stability to different batches of enzymes and plasmids.There were 7 among 50 of diseased pig samples were SVA positive by multiple nano-PCR detec-tion method,and 5 out of 50 of diseased pig samples were SVA positive by ordinary single PCR method.Moreover,no VSNJV and VSIV were detected by the two methods.In conclusion,this es-tablished multiple nano-PCR detection method has higher specificity and sensitivity in the detec-tion of SVA,VSNJV and VSIV.And this study could provide technical support for the rapid differ-ential diagnosis,prevention and control of swine viral vesicular diseases in clinical settings.

Oropharyngeal squamous cell carcinoma(OPSCC)is a malignant tumor originating from the squamous epithelium of the oro-pharyngeal mucosa,accounting for more than 90％of oropharyngeal malignancies.In recent years,human papillomavirus(HPV)infec-tion has become one of the primary etiological factors of oropharyngeal squamous carcinoma.The incidence of HPV-associated oropharyn-geal squamous carcinoma has been rising annually,with a noticeable trend toward younger populations,posing a significant threat to hu-man health.Due to the distinct biological behavior and clinical characteristics of HPV-associated oropharyngeal squamous carcinoma com-pared to its non-HPV-related counterpart,the diagnostic and treatment strategies for oropharyngeal squamous carcinoma have undergone substantial changes.Prevention and screening for oropharyngeal squamous carcinoma are of critical importance.The diagnostic and treat-ment process involves multi-disciplinary collaboration,including oral and maxillofacial surgery,otolaryngology,head and neck surgery,oncology,radiology and pathology.Based on evidence from clinical practice,a comprehensive,integrated diagnostic and therapeutic ap-proach has been established,centered around the concept of"prevention,screening,diagnosis,treatment,and rehabilitation",covering the entire patient lifecycle and providing a valuable reference for clinical practice.

Objective To establish a highly sensitive and specific method for detecting human DNA based on real time quantitative PCR(qPCR)technique for the rapid detection of potential DNA con-tamination sources in DNA laboratories.Methods Primers and probes were designed with Primer Ex-pressTM software using the reference sequence of human 18S rRNA gene as a template,and the opti-mal prime-probe combination was screened by matrix method.The PCR products of the target se-quence of human 18S rRNA gene were used to construct the plasmid,and a plasmid standard was used to draw the standard curve of the qPCR system.According to the Minimum Information for Pub-lication of Quantitative Real-time PCR Experiments(MIQE)guidelines,the specificity,sensitivity,re-peatability and application effect of the qPCR system were evaluated.Results The sensitivity of the qPCR system established in this study was 5.3×10-5 ng/μL,which showed good specificity for human DNA samples.The correlation coefficient of the qPCR system was-0.999,and amplification efficiency was 100％.Both the intra-batch and inter-batch variation coefficients were less than 2％.Conclusion The established human DNA detection method based on qPCR technique has good specificity,high sen-sitivity,and robust stability.It can be used for rapid detection of DNA contamination and daily moni-toring of the accumulated human DNA in the laboratory environment.