Objective To design and synthesize novel Hsp90 inhibitors with dual functions of synergistically enhancing the antifungal activity of fluconazole （FLC） against drug-resistant fungi and anti-tumor activity based on the Hsp90 inhibitor Ganetespib. Methods The previous research found that Ganetespib had a good synergistic anti-resistant fungal activity with FLC, with a fractional inhibitory concentration index （FICI） of 0.023 to 0.039. In this study, structural modifications were made to Ganetespib by replacing its indole ring with a phenyl ring containing different substituents to design and synthesize a series of new compounds. The in vitro synergistic anti-resistant fungal activity against C. albicans 0304103 in combination with FLC, anti-tumor activity （against HEL, HL60 and A549 cells）, and Hsp90α inhibition activity were determined to explore their structure-activity relationship and mechanism of action. Results The chemical structures of 19 new compounds were confirmed by ¹H NMR, ¹³C NMR and HRMS. Most of the compounds exhibited strong Hsp90α inhibitory activity, good synergistic activity against drug-resistant fungi in combination with FLC and anti-tumor activity. The substitution of electron-donating groups on the benzene ring was beneficial to enhancing the synergistic activity against drug-resistant fungi in combination with FLC. Among them, compounds F3 and F5 showed excellent synergistic activity against drug-resistant fungi in combination with FLC （FICI were both 0.047） and anti-tumor activity （IC₅₀ were 0.025 to 0.15 μmol/L and 0.021 to 0.23 μmol/L respectively）, and could down-regulate the expression levels of drug resistance genes and efflux pump genes in fungi, inhibit the formation of fungal biofilms, and arrest the cell cycle of HEL cells at G0/G1 phase. Conclusion The novel Hsp90 inhibitors such as F3 and F5 could both effectively exert the dual activities of synergizing with FLC to combat drug-resistant fungi and fight against tumors, which provided a new idea for the development of new drugs with dual functions of synergizing with FLC to combat drug-resistant fungi and fight against tumors.

ObjectiveTo collate and compare the characteristics and differences in the methods for constructing animal models of different subtypes of gastroesophageal reflux disease （GERD） based on literature， providing a reference for researchers in this field regarding animal model construction. MethodsExperimental studies related to GERD including reflux esophagitis （RE）， nonerosive reflux disease （NERD） and Barrett's esophagus （BE） model construction from January 1， 2014 to January 27， 2024， were retrieved from databases such as CNKI， Wanfang， VIP， Web of Science， and Pubmed. Information on animal strains， genders， modeling methods including disease-syndrome combination models， modeling cycles were extracted； for studies with model evaluation， the methods of model evaluation were also extracted； then analyzing all those information. ResultsA total of 182 articles were included. SD rats were most frequently selected when inducing animal models of RE （88/148， 59.46%） and NERD （9/14， 64.29%）. For BE， C57BL/6 mice were most commonly used （11/20， 55.00%）. Male animals （RE： 111/135， 82.22%； NERD： 11/14， 78.57%； BE： 10/12， 83.33%） were the most common gender among the three subtypes. The key to constructing RE animal models lies in structural damage to the esophageal mucosal layer， gastric content reflux， or mixed reflux， among which forestomach ligation + incomplete pylorus ligation （42/158， 26.58%） was the most common modeling method； the key to constructing NERD animal models lies in micro-inflammation of the esophageal mucosa， visceral hypersensitivity， and emotional problems， and intraperitoneal injection of a mixed suspension of ovalbumin and aluminum hydroxide combined with acid perfusion in the lower esophagus （8/14， 57.14%） was the most common modeling method； the key to constructing BE animal models lies in long-term inflammatory stimulation of the esophageal mucosa and bile acid reflux， and constructing interleukin 2-interleukin 1β transgenic mice （7/25， 28.00%） was the most common modeling method. Adverse psychological stress was the most common method for inducing liver depression. ConclusionsThe construction key principles and methodologies for RE， NERD， and BE animal models exhibit significant differences. Researchers should select appropriate models based on subtype characteristics （e.g.， RE focusing on structural damage， NERD emphasizing visceral hypersensitivity）. Current studies show insufficient exploration of traditional Chinese medicine disease-syndrome combination models. Future research needs to optimize syndrome modeling approaches （e.g.， composite etiology simulation） and establish integrated Chinese-Western medicine evaluation systems to better support mechanistic investigations of traditional Chinese medicine.

ObjectiveThis study aims to explore the effects of Huangqin Qingre Chubi capsules-containing serum on the expression of CircRNA_0001543/nuclear factor-kappa B （NF-κB） in co-cultured peripheral blood mononuclear cells （PBMCs） and human fibroblast-like synoviocytes （FLSs） from patients with ankylosing spondylitis （AS）. MethodsVenous blood was collected from patients with AS to isolate PBMCs. FLSs were co-cultured with AS patients' PBMCs， and FLSs were harvested after co-culture for subsequent experiments. The normal control group consisted of normal FLSs， while the model group comprised co-cultured AS PBMCs and FLSs to simulate AS pathology. The Huangqin Qingre Chubi capsules group involved adding Huangqin Qingre Chubi capsules-containing serum to the co-cultured cells（6.48 g·kg^-1）. To investigate the effect of HQC-containing serum on the viability of co-cultured cells， and the experiment was divided into the following groups based on the dilution concentration： blank group， 10% HQC group， 20% HQC group， and 30% HQC group.To study the influence of the optimal concentration of HQC-containing serum on cytokine and pathway indicators in each group， the experiment was divided into three groups： normal group， model group， and optimal concentration HQC-containing serum group.For the validation of the transfection efficiency of the CircRNA_0001543 interference plasmid， the experiment was divided into the following groups： blank group， si-NC group （with transfection reagent）， si-circ_0001543-1 group （with transfection reagent and interference plasmid No. 1 targeting circ_0001543）， si-circ_0001543-2 group （with transfection reagent and interference plasmid No. 2 targeting circ_0001543）， and si-circ_0001543-3 group （with transfection reagent and interference plasmid No. 3 targeting circ_0001543）.For the validation of the transfection efficiency of the CircRNA_0001543 overexpression plasmid， the experiment was divided into the following groups： blank group， OE-NC group （with transfection reagent）， and OE-circ_0001543 group （with transfection reagent and overexpression plasmid targeting circ_0001543）.To study the effects of CircRNA_0001543 interference/overexpression on cytokine and pathway indicators in each group， the experiment was divided into the following groups： si-NC group， si-CircRNA_0001543 group， OE-NC group， and OE-CircRNA_0001543 group. Enzyme-linked immunosorbent assay （ELISA） was used to detect levels of interleukin-1β （IL-1β）， IL-10， IL-37， and tumor necrosis factor-α （TNF-α）. Real-time quantitative polymerase chain reaction （Real-time PCR） was utilized to measure the expression of CircRNA_0001543， IκBα， and NF-κB p65. ResultsAfter 48 hours， 30% Huangqin Qingre Chubi Capsules-containing serum significantly inhibited the proliferation of co-cultured PBMCs and FLSs， which was determined to be the optimal experimental drug-containing serum concentration. Compared with those in the normal group， the expressions of NF-κB p65 mRNA， IκBα mRNA， IL-1β， and TNF-α in the model group were significantly increased （P<0.01）， while the expressions of CircRNA_0001543 mRNA， IL-10， and IL-37 were significantly decreased （P<0.01）. Compared with those in the model group， the expressions of NF-κB p65 mRNA， IκBα mRNA， IL-1β， and TNF-α in the Huangqin Qingre Chubi Capsules-containing serum group were significantly decreased （P<0.05）， and the expressions of CircRNA_0001543 mRNA， IL-10， and IL-37 were significantly increased （P<0.05）， with the most prominent changes in the 30% drug-containing serum group （P<0.01）. Compared with that in the si-NC group， the expression of CircRNA_0001543 was significantly reduced in the si-CircRNA_0001543 group （P<0.01）. Compared with that in the OE-NC group， the expression of CircRNA_0001543 was significantly increased in the OE-CircRNA_0001543 group （P<0.01）， indicating that the si-CircRNA_0001543 and OE-CircRNA_0001543 plasmids were successfully transfected. Based on the optimal drug-containing serum of Huangqin Qingre Chubi Capsules， si-CircRNA_0001543 transfection led to significantly increased expressions of NF-κB p65 mRNA， IκBα mRNA， IL-1β， and TNF-α and decreased the expressions of IL-10 and IL-37 （P<0.01）. In contrast， OE-CircRNA_0001543 transfection significantly decreased the expressions of NF-κB p65 mRNA， IκBα mRNA， IL-1β， and TNF-α （P<0.01） and increased the expressions of IL-10 and IL-37 （P<0.01）. ConclusionHuangqin Qingre Chubi capsules-containing serum can improve immune inflammation in AS by increasing the expression of CircRNA_0001543， regulating the NF-κB pathway， suppressing pro-inflammatory cytokines， and enhancing anti-inflammatory cytokine expression.

Objective: To investigate the effects of different storage temperatures and durations on the activities of coagulation factor Ⅷ (Factor Ⅷ, FⅧ) and coagulation factor Ⅸ (Factor Ⅸ, FⅨ) after whole blood collection, so as to provide data support for the optimal storage conditions. Methods: A total of 16 mL of whole blood was collected from each of the 20 healthy volunteers at our blood center and aliquoted into 8 sodium citrate anticoagulant tubes. Two tubes were immediately centrifuged for the measurement of FⅧ and FⅨ activity levels. The remaining 6 tubes of whole blood were respectively stored under room temperature and low-temperature conditions. At 2, 4, and 6 h, the whole blood samples were centrifuged and analyzed for FⅧ and FⅨ activity levels. The mean values of the two immediately tested tubes were used as the control group, while other tubes were designated as the experimental groups for comparison. Statistical analysis was performed using SPSS 26.0. Results: The activity of FⅧ in whole blood remained stable after 4 hours of storage at both room temperature and low temperature (116.53±25.95 vs 125.22±27.33, 109.77±23.23 vs 125.22±27.33) (P>0.05 for both). However, by 6 hours, FⅧ activity showed a statistically significant decline compared to the control group (108.65±22.92 vs 125.22±27.33, 100.46±20.19 vs 125.22±27.33) (P<0.05 for both), though the room temperature group results were closer to the control values. The activity of FⅨ in whole blood remained stable after 6 hours of storage under both conditions (97.14±19.48 vs 96.76±19.67, 97.10±17.45 vs 96.76±19.6) (P>0.05 for all comparisons). Conclusion: For whole blood samples after collection, storage at either room temperature or low temperature for up to 4 hours does not compromise the accuracy of test results. When stored for 6 hours, FⅨ activity remains stable, whereas FⅧ activity decreases significantly. Notably, FⅧ activity demonstrates better stability at room temperature than under low-temperature conditions within the 6-hour storage.

Although symptomatic treatment is widely applied in clinical practice， it is often regarded as a relatively low-level therapeutic method. Based on Traditional Chinese Medicine （TCM） state theory， the macroscopic， mesoscopic， and microscopic characterization parameters of TCM symptomatology are horizontally integrated， the full life cycle of states （pre-disease， incipient disease， manifest disease， post-disease） is vertically covered， and the cognitive system of "symptoms" is reconstructed from multiple dimensions. Accordingly， the application approach of symptomatic treatment at different state stages is proposed： implementing preventive intervention in the pre-disease state， strengthening the interception of disease progression in the incipient disease state， regulating dynamic development and treatment in the manifest disease state， and formulating a staged diagnosis and treatment strategy which focuses on functional rehabilitation in the post-disease state.

AIM: To analyze the recovery of stereopsis and its influencing factors in children with intermittent exotropia(IXT)after binocular vision training.METHODS: A total of 166 cases of IXT children who were treated in our hospital from October 2021 to October 2023(2 cases lost their follow-up, and 164 cases were finally included)were included as the research object, taking 36 cases with no stereopsis after binocular vision training in eye position correction surgery as no stereopsis group, and other 128 cases as stereopsis group. All the children underwent eye position correction surgery under general anesthesia, and all received binocular vision training for 6 mo after surgery. The recovery of stereoscopic vision of IXT children after binocular vision training was counted, and the influencing factors of stereoscopic vision recovery of IXT children after binocular vision training were analyzed by single factor and multi-factor Logistic regression analysis.RESULTS: The incidence of postoperative no stereopsis was 22.0%. The proportion of children with an age ≥9 years old, course of disease ≥1 a and anisometropia in the group without stereoscopic vision after operation was larger than the group with stereoscopic vision(all P<0.05). Multivariate Logistic regression analysis showed that the course of disease ≥1 a, age ≥9 years old and anisometropia were independent influencing factors for the recovery of stereoscopic vision in IXT children after binocular vision training(OR=1.470, 1.626, 1.539, all P<0.05).CONCLUSION: Age ≥9 years old, course of disease ≥1 a, and anisometropia are the independent influencing factors of stereopsis recovery of IXT children after binocular vision training. Therefore, targeted intervention measures can be given to high-risk children to improve the stereopsis recovery of IXT children after binocular vision training.

Objective: To construct prediction models of female stress urinary incontinence (SUI), and evaluate the efficacy of each model, so as to provide reference for the early diagnosis of SUI. Methods: Female SUI patients treated in our hospital during Oct. 2019 and Oct. 2023 and healthy women undergoing physical examination during the same period were involved. Women 42 days after delivery were included in the postpartum group (n=611), and perimenopausal and postmenopausal women were included in the non-postpartum group (n=409). The number of random seeds was set and the participants were divided into the training and verification sets in a ratio of 7∶3. Relevant clinical data were collected, and meaningful variables were screened using single factor and Lasso regression, which were then incorporated into the K-nearest neighbor method (KNN), support vector machine (SVM)，decision tree (DT) and random forest (RF) algorithms. The sensitivity, specificity, accuracy and area under the receiver operating characteristic curve (AUC) of the models were calculated to screen out the optimal model. Results: There were 352 SUI patients (57.6%) in the postpartum group. According to single factor and Lasso regression, significant variables included age, body mass index (BMI), maximum rapid muscle stage, parity, bladder neck mobility (BND), urethral rotation angle (URA), lateral perineal incision, past incontinence, and constipation. In the verification set, the AUC of KNN，SVM，DT and RF models were 0.881，0.878，0.750 and 0.905,respectively; the AUC, accuracy, F1 index and Kappa value of RF model were the largest. In the non-postpartum group, there were 260 SUI patients, accounting for 63.6%. The significant variables were age，BMI, maximum value and recovery time of fast muscle stage, mean value of slow muscle stage, post-resting stage variability, vaginal delivery, past incontinence, and constipation. In the verification set, the AUC of KNN,SVM，DT and RF models were 0.819,0.805，0.603 and 0.830, respectively; the AUC, accuracy, Kappa value of the RF model were the largest. Conclusion: This study successfully established 4 prediction models for the incidence of SUI in women at 42 days postpartum, perimenopausal and postmenopausal women based on machine learning. Among them, the model adopting the RF algorithm had the best prediction efficiency.

[摘 要] 目的：探讨烯醇化酶1（ENO1）对胃癌细胞增殖、迁移及侵袭的影响及其分子机制。方法：通过WB法检测ENO1在人胃癌细胞HGC27、MKN-45、N-87、MGC803、BGC-823及人胃黏膜上皮细胞GES-1中的表达水平，利用CRISPR、过表达等基因编辑工具分别构建ENO1敲低及敲低—恢复表达细胞株，将MKN-45和BGC-823细胞分别分组为Ctrl组、ENO1 KD组、ENO1 KD-OE组。通过克隆形成实验、EdU染色、划痕实验、Transwell实验、流式细胞术检测敲低或敲低—恢复表达ENO1对胃癌细胞增殖、迁移、侵袭及凋亡的影响。构建胃癌细胞裸鼠移植瘤模型，通过小动物活体成像技术及肿瘤组织块测量观察ENO1对肿瘤生长的影响。在MKN-45细胞中通过RNA干扰技术沉默ENO1，利用RNA免疫共沉淀-转录组测序（RIP-Seq）结合生物信息学分析技术鉴定ENO1下游的靶基因，分析ENO1调控胃癌细胞增殖、迁移及侵袭的分子机制。结果：ENO1在胃癌细胞中表达水平均显著升高（P < 0.01或P < 0.001）。在MKN-45和BGC-823细胞中敲低ENO1可显著降低胃癌细胞的增殖、迁移、侵袭能力并促进细胞凋亡（P < 0.001或P < 0.000 1）；回复实验结果显示，恢复ENO1的表达后，胃癌细胞的增殖、迁移、侵袭能力均显著提高并减少细胞凋亡（P < 0.05或P < 0.01或P < 0.001或P < 0.000 1）。裸鼠荷瘤实验结果表明，敲低ENO1的表达可以显著抑制裸鼠移植瘤的生长（P < 0.000 1）。与ENO1蛋白具有相互作用的差异基因主要富集于RNA剪切相关通路，且ENO1蛋白与PKM基因具有相互作用，胃癌组织中ENO1与PKM2基因表达事呈正相关（r = 0.886）。结论：ENO1在胃癌细胞中呈高表达，其通过与PKM的前体mRNA相互作用影响PKM的RNA剪接过程，进而调控PKM2的表达水平并促进胃癌的进展。

ObjectiveTo investigate the potential machanism of Xiaozhong Zhitong Mixture （消肿止痛合剂， XZM） in the treatment of diabetes foot ulcer （DFU）. MethodsFifty SD rats were randomly divided into blank group， model group， XZM group， inhibitor group， XZM plus inhibitor group （combination group）， with 10 rats in each group. Except for the blank group， rats were fed with high-sugar， high-fat， high-cholesterol diet， intraperitoneally injected with streptozotocin， and subjected to skin defect to establish DFU model. After successful modeling， the XZM group and the combination group were given 1 ml/（100 g·d）of XZM by gavage， while the blank group， model group， and inhibitor group were all given an equal volume of 0.9% sodium chloride injection by gavage. Thirty minutes later， the inhibitor group and the combination group were intraperitoneally injected with 5 mg/（kg·d） of Notch1 inhibitor DAPT. All groups were treated once a day. After 14 days of administration， the skin tissue from the dorsal foot of the blank group rats and wound tissue from the other groups were collected. The pathological changes of granulation tissue in the wound were detected using hematoxylin eosin （HE） staining. The microvascular density （MVD） in wounds was detected through immunohistochemical staining. Real time fluorescence quantitative polymerase chain reaction （RT-PCR） and western blotting were used to detect the mRNA and protein levels of Notch1 homolog （Notch1）， Delta-like ligand 4 （Dll4）， Delta-like ligand 4 （VEGF）， and angiopoietin 2 （Ang-2）， respectively. ResultsHistological results showed that the epidermal structure in the dorsal foot skin tissue of the rats in the blank group was intact. In the wound tissue of the model group， the epidermis exhibited excessive keratinization， vacuolar cytoplasm， and a large number of inflammatory cells infiltrating the tissue， while in the XZM group， a large amount of scab formation was observed in the epidermis， with no significant inflammatory cell infiltration and a noticeable increase in fibroblasts. In the combination group and the inhibitor group， partial epidermal scab formation was observed in the wound tissue with a small amount of inflammatory cell infiltration. Compared to those in the blank group， the MVD in the wound tissue increased in the model group， as well as the mRNA expression and protein levels of Notch1 and Dll4， while VEGFA and Ang-2 mRNA expression and protein levels significantly decreased （P＜0.05 or P＜0.01）. Compared to those in the model group， the MVD in the wound tissue of all medication groups significantly increased， and the mRNA and protein levels of Notch1 and Dll4 decreased， while VEGFA and Ang-2 mRNA expression and protein levels increased （P＜0.05 or P＜0.01）. Compared to the XZM group， the inhibitor group and the combination group showed decreased MVD in wound tissue， increased Notch1 and Dll4 mRNA and protein levels， and decreased expression of VEGFA and Ang-2 mRNA and proteins （P＜0.05 or P＜0.01）. ConclusionXZM can effectively promote wound healing in DFU rats， and its mechanism of action may be related to the inhibition of Dll4/Notch1 signaling pathway in the wound tissue， therey promoting angiogenesis.

The field of post-marketing benefit-risk assessment for traditional Chinese medicine（TCM） is still in its nascent stage， lacking a universally accepted and cohesive evaluation framework and standards. This study presented a strategy developed for the benefit-risk assessment of post-marketing of TCM， and explored the critical techniques and specific implementation steps involved in the assessment process. Initially， appropriate qualitative assessment frameworks and quantitative analysis models were selected for the integrated qualitative and quantitative benefit-risk assessment. Subsequently， key technologies were outlined， including the establishment of a benefit-risk indicator system， the assignment of indicator weights， and the definition of criteria attributes. Furthermore， the implementation steps were elaborated， which involved defining decision-making issues， data collection， evaluation methodologies， variability factors， and sensitivity analysis. Finally， a case study of the benefit-risk assessment of a TCM injection for hepatitis B treatment was conducted to validate the feasibility of the proposed strategy. The objective of this research was to provide theoretical support and practical references for the development of a comprehensive post-marketing benefit-risk assessment system for TCM.