To systematically analyzed the reporting status of core elements in publicly available clinical practice guideline(hereafter referred to as "guideline") protocols published domestically and internationally over the past decade, identified existing problems, and provided evidence to inform the standardized writing and publication of future guideline protocols.

Objective To explore the prevalence of Helicobacter pylori (Hp) infection and its association with dietary and lifestyle habits among the adult physical examination population in Xuzhou area. Methods Retrospectively selected the physical examination population who underwent HP testing at our hospital's physical examination center from May 2021 to December 2023 as the research object. The prevalence of Hp infection in the population was analyzed based on the physical examination results. A questionnaire survey was used to collect information on the eating and living habits of all study subjects. Logistic regression was used to analyze the relationship between eating and living habits and Hp infection. Results A total of 1 354 physical examination people were included in the study, and the Hp infection rate was 37.30% (505/1354). The difference in Hp infection rates among people of different age groups is statistically significant (P<0.05), with the middle-aged population (41-59 years old) having the highest Hp positive infection rate (45.38%).High salt (41.11%), hot diet (40.56%), history of smoking (45.23%) and drinking (43.80%), less consumption of fruits and vegetables (43.73%), irregular exercise (41.29%), irregular diet People who frequently eat out (43.56%) and eat out frequently (42.57%) have a higher Hp infection rate (P<0.05).After adjusting for demographic factors such as gender, age, place of residence and education level, multivariate Logistic regression results showed that high-salt diet (OR=3.975, 95%CI: 2.670-5.917) and hot diet (OR=3.357, 95%CI: 2.291-4.919), smoking (OR=1.458, 95%CI: 1.082-1.964), drinking alcohol (OR=1.654, 95%CI: 1.279-2.138), eating fruits and vegetables (OR=1.759, 95%CI: 1.345-2.301), regular exercise (OR=1.822, 95%CI: 1.371-2.421), regular diet (OR=1.893, 95%CI: 1.391-2.575), eating out (OR=1.690, 95%CI: 1.277-2.237) were associated with the risk of Hp infection (P<0.05). Conclusion The positive infection rate of Hp among the physical examination population in Xuzhou is slightly lower than the average epidemic level in China. Cultivating healthy eating and living habits can effectively reduce the risk of Hp infection.

Animal experimentation constitutes a critical link between basic research and clinical application, making its research quality and translational efficiency paramount. Although considerable progress has been made in standardizing operational procedures and ethical guidelines, the standardization of outcome evaluation systems has significantly lagged, creating a key bottleneck that constrains the quality of biomedical research and evidence synthesis. This deficiency is manifested by pronounced heterogeneity in outcome selection across similar studies, incomplete methodological reporting, and disparate criteria for result interpretation, which severely impairs the comparability of findings and the evidence integration. To cope with this challenge, this paper systematically introduces a mature methodological tool from clinical research–the core outcome set (COS)–and explores its construction strategies and application potential in the field of animal experimentation. Given the extensive diversity of animal experiments, a pragmatic strategy of "focusing on key areas, implementing phased pilots, and promoting gradual expansion" should be adopted. This approach prioritizes the development of domain-specific COS for disease areas characterized by high research volume, urgent translational needs, and well-established animal models. A multi-source integration pathway for COS development is detailed, comprising systematic literature searches, methodological appraisals, and expert consensus, with the feasibility of leveraging artificial intelligence (AI) to enhance efficiency also being examined. The development and promotion of such COS are not intended to restrict scientific exploration; rather, they aim to establish a new, tiered evaluation paradigm consisting of "core outcomes" (mandatory), "recommended outcomes" (encouraged), and "exploratory outcomes" (optional). This framework is expected not only to enhance research quality through standardization and to adhere to the "3R" principles but also to accelerate the accumulation of high-quality evidence. This, in turn, provides a solid foundation for higher-level evidence synthesis, ultimately facilitating the effective translation of basic research findings into clinical practice and providing an essential methodological framework for scientific advancement in relevant disciplines.

Objective To express the capsid(Cap) protein of porcine circovirus 2(PCV2) by insect-baculovirus expression system, evaluate its immunogenicity as a vaccine candidate, and explore whether it can induce humoral and cellular immune responses in BALB/c mice.Methods Following codon optimization, the PCV2 Cap gene was cloned into pFastBac Dual vector and transformed into E.coli DH10 Bac cells. After Bacmid extraction, PCR amplification was carried out with M13 universal primers. The appropriately identified Bacmid was then transfected into Sf9 cells to express Cap protein, and the protein was quantified. Forty-five female BALB/c mice were randomly divided into three groups: PBS control, Cap and commercial vaccine YI Cap group, 15 mice in each group, which were immunized subcutaneously on the back once at 14-day intervals, for a total of three times, with PBS 100 ??L, Cap 10 ??g, and commercial vaccine YI Cap 10 ??g respectively each time. The serum IgG level of mice was measured by ELISA, the cellular immune response type was analyzed by flow cytometry, and the histocompatibility of the subunit vaccine was observed by H&E staining.Results The insect-baculovirus expression system effectively expressed the target protein, and the Cap protein with enhanced green fluorescent protein(eGFP) colocalized with the cell's plasma membrane. Transmission electron microscopy revealed that the Cap protein could self-assemble to produce virus-like particles(VLPs) before constructing another recombinant vector sans the eGFP fluorescent flag. SDS-PAGE and Western blot analysis revealed the conspicuous bands at the desired area. The IgG content of Cap vaccine candidate mice reached the highest level at 42 days. The levels of IFNγ and IL-4 were(27. 75 ± 0. 99) and(17. 63 ± 1. 13) pg/mL, respectively, both of which were significantly higher than those of PBS group(F = 14. 89 and 0. 211 9,P < 0. 05 and < 0. 001, respectively). The stimulation index(SI) of splenic lymphocytes in the Cap group reached 6. 72, which was significantly higher than that in the PBS group(F = 1. 228, P < 0. 001). The Cap vaccine candidate could induce Th1-type cellular immune response and exhibited good histocompatibility.Conclusion The Cap protein was successfully expressed in the insect-baculovirus expression system. The Cap vaccine candidate can induce humoral and cellular immune responses in mice after immunization, which is a potential vaccine candidate for further development of more effective vaccines against PCV-related diseases.

ObjectiveHeadspace solid-phase microextraction-gas chromatography-mass spectrometry（HS-SPME-GC-MS） and GC-triple quadrupole MS（GC-QqQ-MS） in combination with non-targeted and targeted metabolomics were employed to systematically analyze the chemical composition differences of Xihuangwan prepared with natural musk and artificial musk， and establish an identification system for them. MethodsThe volatile components of 9 batches of Xihuangwan samples from 8 manufacturers were analyzed by HS-SPME-GC-MS non-targeted metabolomics， and identified by comparing their MS data with the National Institute of Standards and Technology（NIST） spectral library. Orthogonal partial least squares-discriminant analysis（OPLS-DA） was used to identify differential volatile components of Xihuangwan prepared with natural musk and artificial musk. Additionally， GC-QqQ-MS targeted metabolomics was applied to quantify the levels of α-pinene， β-elemene， muscone， dehydroepiandrosterone， bornyl acetate， and octyl acetate in 27 batches of samples from 9 manufacturers. Cluster analysis， principal component analysis（PCA）， and partial least squares-discriminant analysis（PLS-DA） were conducted to further explore the differences in volatile components between Xihuangwan samples prepared with natural musk and artificial musk. ResultsNon-targeted metabolomics identified 291 volatile compounds in Xihuangwan， including alkanes， esters， alkanes， alcohols， ketones， naphthalenes and others. OPLS-DA analysis revealed distinct separation between Xihuangwan samples containing artificial musk（A1， C1， D1， E1， F1， G1， I1） and those containing natural musk（H1， H3）. A total of 30 differential metabolites were identified. The relative contents of these 30 differential metabolites were visualized using a radar chart， revealing significant differences in the levels of octanol， borneol acetate and muscone. Cluster analysis and PCA results from targeted metabolomics indicated that Xihuangwan could be classified into two distinct groups：one composed of natural musk（H1， H3） and the other of artificial musk， sample H2. PLS-DA identified muscone， octyl acetate， and dehydroepiandrosterone as key differential volatile components. Although no significant difference was observed in the content of octyl acetate between the two groups， statistically significant differences were found for muscone and dehydroepiandrosterone（P<0.05）. ConclusionMuscone and dehydroepiandrosterone can be used for the differentiation of Xihuangwan samples containing natural musk from those containing artificial musk. This study systematically and comprehensively analyzed the differences in the types and contents of major volatile components in Xihuangwan prepared with natural musk and artificial musk， providing a scientific basis for quality evaluation and control of Xihuangwan.

Polycystic ovary syndrome （PCOS） is a common endocrine and metabolic disorder among women of reproductive age. Hyperandrogenism （HA）， one of its core pathological features， is closely associated with the clinical manifestations and metabolic complications of the disease. Current western medical treatments for PCOS-HA mainly include anti-androgen therapy and ovulation induction， such as short-acting oral contraceptives like Diane-35 and Yasmin. However， long-term use of these medications may result in adverse reactions like increasing the risk of liver dysfunction and exacerbating lipid metabolism disorders， with unsatisfactory long-term efficacy when used alone. Traditional Chinese medicine offers unique advantages in the treatment of PCOS-HA due to its holistic approach and multi-target regulatory mechanisms. In the view of traditional Chinese medicine， PCOS-HA is classified under the categories such as "delayed menstruation"， "amenorrhea"， and "infertility"， with kidney deficiency as the root， as well as liver stagnation and spleen deficiency as the manifestations. Phlegm and blood stasis are considered to be intertwined throughout the disease course. Modern studies have shown that traditional Chinese medicine is significantly effective in improving the androgen levels， restoring ovulation， and improving insulin resistance in PCOS-HA patients. Representative prescriptions， such as Erxian Tang， Jiawei Xiaoyaosan， Guizhi Fulingwan， and Cangfu Daotantang， exert therapeutic effects through various mechanisms including regulation of the hypothalamic-pituitary-ovarian axis， reduction of ovarian androgen synthase activity， improvement of insulin signaling pathways， and inhibition of inflammation and oxidative stress， which demonstrates the characteristics of comprehensive treatment with traditional Chinese medicine. Based on the perspectives of etiology and pathogenesis of traditional Chinese medicine， modern medical cognition， typical prescriptions， and action mechanisms， this paper reviewed the research progress of traditional Chinese medicine in the treatment of PCOS-HA， aiming to provide a reference for in-depth research and clinical applications in this field.

ObjectiveTo prepare Mycobacterium tuberculosis ESAT6-Fc fusion protein and evaluate its efficacy and safety on ovalbumin(OVA)-induced allergic rhinitis(AR) in mice, so as to provide experimental basis for the development of novel AR immunothera-peutic agents.MethodsThe recombinant plasmid pcDNA3.1(+)-Rv3875-Fc was transfected into CHO-K1 cells to obtain ESAT6-Fc fusion protein, which was purified by Protein G affinity chromatography. The female C57 BL/6J mice of AR model sensitized by OVA were randomly divided into five groups: normal control, OVA, CpG, ESAT6-Fc + CpG and dexamethasone(DEX) group, six mice each group, which were challenged with atomized OVA after the corresponding preparation was given to the nasal cavity for intervention. HE staining method was used to evaluate the inflammatory infiltration of mouse nasal mucosa, and ELISA method was used to detect the content of histidine(HIS), leukotriene B4(LTB4), IgE, IL-4,IL-13 and IL-17A in the nasal lavage fluid. The structural changes of heart, liver, spleen and kidney tissues were observed by HE staining, and the levels of glutamic oxaloacetic transaminase(AST) and lactate dehydrogenase(LDH) in mouse heart,liver, spleen and kidney were measured by ELISA.ResultsESAT6-Fc fusion protein with high purity(> 95%) was successfully obtained. Compared with the normal control group, the inflammatory cell infiltration in nasal mucosa of the OVA group was significantly aggravated(t = 11. 180, P < 0. 001), and the levels of HIS, LTB4, IgE, IL-4, IL-13 and IL-17A in the nasal lavage fluid significantly increased(t = 3. 843, 4. 237, 5. 996, 3. 561, 4. 544, 5. 773, respectively, each P < 0. 05). Compared with the OVA group, the infiltration of inflammatory cells in nasal mucosa of mice in the ESAT6-Fc + CpG and DEX groups was significantly reduced(t = 4. 472 and 5. 582, respectively, each P < 0. 001), and the levels of HIS, IgE, LTB4, IL-4, IL-13 and IL-17A in the nasal lavage fluid significantly decreased(t = 0. 000-1. 061, each P < 0. 05). There was no significant difference in the levels of HIS, IgE, LTB4, IL-4, IL-13 and IL-17A between the ESAT6-Fc + CpG and DEX groups(t = 0. 048, 0. 000,0. 968, 1. 061, 0. 375 and 0. 638, respectively, each P > 0. 05). Intranasal immunization with ESAT6-Fc did not cause pathological damage to the heart, liver, spleen, and kidney of mice, and the levels of AST and LDH in the above organs showed no significant difference from those in the normal control group(t = 0. 001-1. 036, each P > 0. 05).ConclusionThe prepared ESAT6-Fc fusion protein has high purity, and intranasal immunization with the fusion protein can safely and effectively alleviate OVA-induced AR symptoms in mice, and inhibit the expression of related inflammatory factors, without obvious organ toxicity observed, indicating that ESAT6-Fc fusion protein is expected to become a new type of immunotherapeutic agent for AR.

ObjectiveTo evaluate the safety, efficacy and economy of equine tetanus immunoglobulin F(ab')_2, so as to provide reference for its clinical application.MethodsPatients who were admitted to six research centers for trauma from November 23, 2021 to October 31, 2022 and required a tetanus immunoglobulin injection were selected as the research subjects to observe the positive rate of skin sensitization test of equine tetanus immunoglobulin F(ab')_2 and the incidence of adverse events(AEs) after injection. In addition, one of the study centers was selected to evaluate the incidence of tetanus in patients receiving equine tetanus immune globulin F(ab')_2 injection in this center. Meanwhile, a pharmacoeconomic model per 10 000 patients was constructed to compare the difference in drug costs of switching to human tetanus immunoglobulin(TIG) due to a positive skin sensitization test when patients preferred tetanus antitoxin(TAT) or equine tetanus immunoglobulin F(ab')_2.ResultsA total of 422 patients were enrolled in the study, of whom 419 underwent an equine tetanus immunoglobulin F(ab')_2 skin sensitization test and 416 received a test result, with a positive skin sensitization test rate of 5. 05%(21/416). Among the 395 patients who had a negative skin sensitization test and completed equine tetanus immunoglobulin F(ab')_2 injection, five(1. 27%) patients developed AEs related to equine tetanus immunoglobulin F(ab')_2, mainly including rash and pruritus, and two(0. 51%) patients had grade ≥ 3 AEs related to equine tetanus immunoglobulin F(ab')_2, including one case of immediate anaphylaxis and one case of pruritus. None of the patients developed anaphylactic shock or death related to equine tetanus immunoglobulin F(ab')_2. None of the patients in the selected research center developed tetanus after administration of equine tetanus immunoglobulin F(ab')_2. In addition, the pharmacoeconomic evaluation model showed that equine tetanus immunoglobulin F(ab')_2 had a higher drug economic value compared to TAT and TIG.Conclusion Equine tetanus immunoglobulin F(ab')_2 has shown good efficacy and safety in preventing tetanus in the real world with high pharmacoeconomic value.

Organ preservation after neoadjuvant therapy for esophageal cancer has gained significant attention. While the CROSS trial established neoadjuvant chemoradiotherapy (nCRT) followed by surgery as standard care, approximately 30% of patients achieve pathological complete response (pCR), prompting exploration of active surveillance (AS). The landmark SANO phase Ⅲ trial (2025) demonstrated non-inferior 2-year overall survival (74% AS vs. 71% surgery), with 31% of patients avoiding surgery. Multimodal assessment (endoscopic deep biopsy+endoscopic ultrasound+PET-CT) reduced residual disease misdiagnosis to 10%. The Asian-led NEEDS trial is evaluating definitive chemoradiotherapy with salvage surgery. Although immunotherapy boosts pCR rates to 40%-55%, challenges persist, including 8%-12% false-negative cCR assessments, limited long-term data, and East-West histological disparities. The 2024 NCCN guidelines conditionally recommend AS (Category 2B, prioritized for squamous cell carcinoma), emphasizing centralized implementation. Future directions involve circulating tumor DNA and radiomics for risk stratification to advance precise organ-preserving strategies.

INTRODUCTION: Glaucoma is a leading cause of permanent blindness, often going undetected in its early, asymptomatic stages, especially in older age groups. In Malaysia, glaucoma is a growing public health issue due to an increase in the ageing population. While screening is essential for early glaucoma detection, the most suitable strategy for Malaysia's healthcare system remains unclear. This study explored the perspectives of eye healthcare professionals on the most suitable glaucoma screening strategies for Malaysia. MATERIAL AND METHODS: This qualitative study used semi-structured interviews with 19 eye health professionals (ophthalmologists, optometrists, nurses, ophthalmic technicians, and assistant medical officers) practicing in the Klang Valley. The interviews were conducted face-to-face in their workplace or via a video conferencing platform. All interviews were recorded, transcribed, and analysed using thematic analysis. RESULTS: Six major themes were identified: types of glaucoma screening programmes, accessible screening locations, target screening population, instruments and use of digital technology, trained personnel, and referral criteria. Opportunistic case finding and population-based programmes were identified as the glaucoma screening programmes in which trained personnel conducted screening at accessible locations. Glaucoma screening for high-risk individuals was recommended, focusing on visual acuity testing, tonometry, anterior chamber angle assessment, funduscopy, perimetry, and retinal nerve fibre assessment. A lack of clear referral criteria due to low awareness and poor implementation of existing guidelines was observed. CONCLUSION: Further investigations are required to identify the best combination of components for glaucoma screening. This will enable policymakers to develop an effective glaucoma screening programme in Malaysia.