Background/Aims: Regorafenib is recommended by guidelines and trials as a sequential second-line therapy following progression on first-line sorafenib or lenvatinib in hepatocellular carcinoma (HCC). However, efficacy is limited, highlighting the urgent need to screen suitable patients and develop sensitization strategies. Methods: Acquired sorafenib- or lenvatinib-resistant (SR or LR) HCC cell lines and organoids were established. Genome-wide CRISPR library screen was performed in SR or LR cell strains to identify synthetic lethal targets of regorafenib. RNA-seq and FITC-regorafenib efflux assay were used to elucidate ERBB3-driven downstream signaling. Preclinical mouse models of cell line- and patient-derived xenografts and clinical cohorts of HCC patients were employed to validate the efficacy of ERBB3-guided patient stratification. Results: Screening with CRISPR library, we showed that inhibition of ERBB3 was synthetic lethal with regorafenib in SR or LR cell strains and organoids. Mechanistically, SR or LR triggered feedback activation of ERBB3 signaling and mediated regorafenib efflux via ERBB3-HIF1A-ABCB1 cascade pathway, limiting sensitivity to regorafenib. Moreover, ERBB3-low tumors following SR or LR exhibited significant sensitivity to regorafenib, suggesting its potential as a predictive biomarker to screen optimal candidates for sequential therapy. Seribantumab, an ERBB3-targeting monoclonal antibody, inhibited ERBB3-HIF1A-ABCB1 cascade, and its combination with regorafenib exerted marked synergistic anti-tumor effects on ERBB3-high tumors resistant to sorafenib or lenvatinib both in vitro and in vivo. Conclusions This study revealed that ERBB3 was a key resistance factor driving limited efficacy to sequential regorafenib, but also an effective therapeutic target whose inhibition enhanced regorafenib sensitivity after SR or LR.

Objective To systematically evaluate the efficacy and safety of traditional Chinese medicine (TCM) compound in treating pulmonary nodules, providing evidence-based medical evidence for TCM intervention in pulmonary nodules. Methods Computer search of PubMed, CNKI, Wanfang, VIP, and SinoMed was conducted to select randomized controlled trials (RCTs) of TCM compound intervention in pulmonary nodules, with the retrieval time from the inception to November 29, 2023. The Cochrane bias risk assessment tool was used to evaluate the quality of the included studies, and Review Manager 5.4 was used for Meta-analysis. Results A total of 18 RCTs were included, covering 8 provinces across the country, with a total sample size of 1301 patients. The TCM compounds used in the included studies all incorporated the method of dissolving phlegm and dissipating nodules. There was a high risk of bias uncertainty in the included studies. Meta-analysis results suggested that TCM compound could significantly reduce the diameter of pulmonary nodules [MD=−1.41, 95%CI (−1.70, −1.13), P<0.001], decrease the number of nodules [MD=−0.37, 95%CI (−0.73, −0.01), P=0.05], alleviate clinical symptoms [MD=−4.84, 95%CI (−6.04, −3.64), P<0.001], and improve lung function [forced expiratory volume in one second (FEV1), MD=0.55, 95%CI (0.09, 1.01), P=0.02; FEV1/forced vital capacity, MD=6.12, 95%CI (4.47, 7.78), P<0.001]. However, there was no statistically significant difference in the probability of malignancy between the experimental group and the control group [MD=−0.01, 95%CI (−0.01, 0.00), P=0.09]. Conclusion TCM compound can significantly reduce the diameter of pulmonary nodules, decrease the number of nodules, alleviate clinical symptoms, and improve lung function, but future multicenter, large-sample, high-quality RCTs are still needed to further explore and verify this conclusion.

ObjectiveTo investigate the inhibitory effect of Huoluo Xiaolingdan on triple negative breast cancer （TNBC） in mice through its regulation of TCF1⁺CD8⁺ stem cell-like T cells infiltration. MethodsA mouse model of TNBC was established and the mice were randomly divided into the model group， low-dose （3.9 g·kg^-1）， medium-dose （7.8 g·kg^-1） and high-dose （15.6 g·kg^-1） Huoluo Xiaolingdan groups， and anti-PD-1 antibody treatment group. Each group was given a dose of 0.01 mL·g^-1， while the model group and the anti-PD-1 treatment group were also given an equivalent volume of normal saline. The drug was administered for 21 days. In the anti-PD-1 antibody group， mice were intraperitoneally injected with 100 μg of mouse anti-PD-1 antibody twice a week， for a total of five injections. The tumor volume， survival time and tumor mass were measured at different time points. Hematoxylin-eosin （HE） staining was used to observe the histological changes of the tumor. The expression of CD8⁺T cells and TCF1⁺CD8⁺ stem-like T cells in tumor tissues was detected by immunohistochemistry and immunofluorescence staining. Flow cytometry was used to detect the difference of immune cell subsets in tumors and the expression difference of TCF1⁺CD8⁺ stem cell-like T cells in tumors and peripheral blood. The expression level of PD-L1 in tumor tissues was detected by Western blot. ResultsCompared with model group， the tumor volume and mass of in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group were decreased （P<0.05， P<0.01）. The median survival time of mice in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group was as follows： 27.00 days （95%CI， 0.45-2.65）， 31.00 days （95%CI， 0.32-1.89）， 34.00 days （95%CI， 0.40-2.33）， and 35.00 days （95%CI， 0.42-2.47）. All of them were higher than that of the model group ［24.50 days （95%CI， 0.37-10.5）］. Flow cytometry showed that compared with the model group， the proportion and number of infiltrating CD8⁺ T cells in tumor were increased in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group （P<0.05， P<0.01）， while the proportion of tumor regulatory T cells （Treg） and M2 macrophages decreased （P<0.05）. Compared with the model group， the proportion of IFN-γ⁺CD8⁺ T and GrzB⁺CD8⁺ T cells in tumors in low-， medium- and high-dose Huoluo Xiaolingdan groups and anti-PD-1 group was increased （P<0.01）， and the proportion of TCF1⁺CD8⁺ T cells in tumor and peripheral blood was also increased. Immunofluorescence staining further showed that the number of TCF1⁺CD8⁺ T cells in tumor tissues increased in low-， medium- and high-dose Huoluo Xiaolingdan groups. Western blot analysis showed no significant decrease in the PD-L1 protein expression in tumor tissues between the Huoluo Xiaolingdan groups and the model group. ConclusionHuoluo Xiaolingdan can inhibit TNBC in mice by increasing tumor infiltration of TCF1⁺CD8⁺ stem-like T cells， enhancing CD8⁺ T cell activity， and regulating immune cell subgroups such as M2 macrophages and Treg cells to enhance anti-tumor immunity. This study provides a theoretical basis for the clinical application of Huoluo Xiaolingdan in breast cancer treatment and combination therapy.

OBJECTIVE: To evaluate the efficacy and safety of Shexiang Tongxin Dropping Pill (STDP) in treating stable angina patients with phlegm-heat and blood-stasis syndrome by exercise duration and metabolic equivalents. METHODS: This multicenter, randomized, double-blind, placebo-controlled clinical trial enrolled stable angina patients with phlegm-heat and blood-stasis syndrome from 22 hospitals. They were randomized 1:1 to STDP (35 mg/pill, 6 pills per day) or placebo for 56 days. The primary outcome was the exercise duration and metabolic equivalents (METs) assessed by the standard Bruce exercise treadmill test after 56 days of treatment. The secondary outcomes included the total angina symptom score, Chinese medicine (CM) symptom scores, Seattle Angina Questionnaire (SAQ) scores, changes in ST-T on electrocardiogram and adverse events (AEs). RESULTS: This trial enrolled 309 patients, including 155 and 154 in the STDP and placebo groups, respectively. STDP significantly prolonged exercise duration with an increase of 51.0 s, compared to a decrease of 12.0 s with placebo (change rate: -11.1% vs. 3.2%, P<0.01). The increase in METs was significantly greater in the STDP group than in the placebo group (change: -0.4 vs. 0.0, change rate: -5.0% vs. 0.0%, P<0.01). The improvement of total angina symptom scores (25.0% vs. 0.0%), CM symptom scores (38.7% vs. 11.8%), reduction of nitroglycerin consumption (100.0% vs. 11.3%), and all domains of SAQ, were significantly greater with STDP than placebo (all P<0.01). The changes in Q-T intervals at 28 and 56 days from baseline were similar between the two groups (both P>0.05). Twenty-five participants (16.3%) with STDP and 16 (10.5%) with placebo experienced AEs (P=0.131), with no serious AEs observed. CONCLUSION STDP could improve exercise tolerance in patients with stable angina and phlegm-heat and blood stasis syndrome, with a favorable safety profile. (Registration No. ChiCTR-IPR-15006020).
Humans ; Double-Blind Method ; Drugs, Chinese Herbal/adverse effects* ; Male ; Female ; Middle Aged ; Angina, Stable/physiopathology* ; Aged ; Syndrome ; Treatment Outcome ; Placebos ; Tablets

Objective:To investigate the safety and efficacy of urinary microbiota transplantation（UMT）in treating interstitial cystitis（IC）.Methods:A retrospective analysis of the clinical data of three patients with IC treated with UMT at the Central Hospital of Jiangnan University from May 2022 to August 2024. Three women（45，62，79 years）presented with urinary frequency（10 - 90 min intervals），urgency，dysuria，lower abdominal pain，and non-organic sleep disorder，with nocturia（2 - 15 episodes）causing sleep difficulty. Disease durations were 3，6，and 21 years. Prior antibiotic therapy failed. Preoperative urinalysis/culture（x2）ruled out bacterial cystitis，and diagnosed as interstitial cystitis（IC），with one of transient positive urine culture. Preoperative scores for case 1 were self-rating depression scale（SDS）of 49，self-rating anxiety scale（SAS）of 31，interstitial cystitis symptom index（ICSI）of 5，interstitial cystitis problem index（ICPI）of 2，brief urological problem scale of 50，genitourinary pain assessment scale of 10，and pain catastrophizing scale（PCS）of 0. For case 2，the scores were 60 of SDS，66 of SAS，9 of ICSI，11 of ICPI，50 of brief urological problem scale，28 of genitourinary pain assessment scale，and 34 of PCS. For case 3，the scores were 44 of SDS，48 of SAS，11 of ICSI，5 of ICPI，33 of brief urological problem scale，27 of genitourinary pain assessment scale，and 21 of PCS. Preoperative bladder hydro-distention showed mucosal hemorrhage including central block hemorrhage（Case 1），numerous spots（Case 2），and distinct sheets（Case 3）. Preoperative urine 16S rRNA sequencing revealed low diversity，increased Gardnerella/ Bacteroides（opportunistic），and decreased Bacillus/ Streptococcus（beneficial）. Two healthy young female donors had normal comprehensive tests covering blood/urine/stool，coagulation，CRP，immunity，liver/kidney function，lipids，infectious diseases，hormones，tumor markers，urine culture，TOUCH，and expanded quantitative urine culture. Donor urine 16S rRNA showed low pathogenic（ Gardnerella/ Bacteroides）abundance. Donor midstream morning urine was catheter-collected，centrifuged，and the bacterial pellet resuspended in saline. Recipients underwent bladder irrigation pre-UMT. On UMT day，donor bacterial suspension was instilled via catheter with adverse event monitoring. Follow-up included clinic visits at 1 month and 1 year for symptom assessment，scale scoring，cystoscopy evaluating mucosal inflammation，hydrodistension checks，and telephone tracking of urinary symptoms every 2 - 3 months. Prognosis was assessed by symptom relief，scale score reduction，and mucosal recovery. Results:No adverse reactions occurred within 24 hours post-UMT in all 3 cases. Two patients showed same-day urinary urgency reduction；three reported relief from urinary frequency/urgency on postoperative day 7，with 2 - 3 hour daytime intervals and 0 - 5 nocturnal voids. Two patients experienced lower abdominal pain alleviation. Postoperative Week 1 urinalysis and urine cultures revealed no abnormalities. The 16S rRNA test showed a decrease in the abundance of harmful bacteria（e.g.， Cupriavidus，Ureaplasm，Mycobacterium，Pseudomonas）and an increase in the abundance of beneficial bacteria（ Dietzia，Halomonas，and Streptococcus），and the overall urobacterial structure was significantly improved and similar to that of the donor. Case 1 was followed up for 20 months，and the lab tests were normal，with SDS scales of 38，SAS of 20，ICSI of 3，ICPI of 2，brief urological problem scale of 44，genitourinary pain assessment scale of 10，PCS of 0，at 9 months post-op follow-up，indicating physical and mental improvement. The bladder hydro-distention revealed intact mucosa with only mild inflammation，which improved versus preoperative situation. Case 2 was followed up for 15 months，and the lab tests were normal，with SDS scales of 44，SAS of 34，ICSI of 4，ICPI of 2，brief scale of urinary problems of 0，genitourinary pain assessment scale of 9，PCS of 7，at 2 months post-op follow-up，showing improved anxiety/depression and quality of life. The hydro-distention showed decreased scattered hemorrhagic dots and mucosal inflammation. Case 3 was followed up for 13 months，with an increased leukocytes of urine and the other being normal for lab tests，and SDS scales of 35，SAS of 46，ICSI of 13，ICPI of 13，brief scale of urinary problems of 10，genitourinary pain assessment scale of 33，PCS of 11，at 13 months post-op follow-up，indicating physical and mental improvement. The hydro-distention revealed mucosal congestion，marked submucosal vasodilation，and inflammation decreased compared with preoperative situation. Conclusions:UMT alleviates urinary frequency，urgency，and pain in IC patients with sustained effects，significantly improves urine microecology，and shows no adverse events，positioning it as a viable intervention for IC.

The “Technical Guideline for Epidemic Prevention and Control Disinfection in Large-Scale Events”(hereinafter referred to as “the Guideline”), organized and compiled by the National Disease Control and Prevention Administration, was officially released in April 2024. This guideline aims to ensure the effective implementation of large-scale group activities, mitigate the impact of infectious disease outbreaks on such events, and maintain hygiene and safety standards at event venues. During the compilation process, data were systematically collected in alignment with epidemic prevention requirements and disinfection principles, incorporating research findings from domestic and international disinfection practices. Information was gathered through field investigations, expert consultations in epidemiology and disinfection, and roundtable discussions with representatives from organizations responsible for disinfection operations at large-scale events, thereby ensuring the scientific rigor and practical applicability of the content. The Guideline provides comprehensive technical disinfection guidance for relevant authorities and event organizers, addressing critical aspects such as disinfection protocols, operational principles, emergency response strategies, and technical specifications. By standardizing hygiene assurance measures for large-scale events, including considerations of participant demographics, venue characteristics, and event scale, the guideline establishes a framework to proactively minimize the risk of infectious disease transmission.

Objective To analyze the stimulation parameters and acupoint selection law of electroacupuncture in the treatment of cervical spondylotic radiculopathy(CSR)through data mining.Methods The clinical research literature about electroacupuncture in the treatment of CSR was retrieved from CNKI,Wanfang Data,VIP,PubMed and Web of Science from the establishment of the databases to September 2024.According to the inclusion and exclusion criteria,the literature was screened and the stimulation parameters and acupoint prescriptions were extracted.The association rule analysis was performed using SPSS Moderler 18.0 software,and clustering analysis was performed using SPSS Statistics 27.0 software.Results A total of 122 acupoint selection prescriptions for electroacupuncture treatment of CSR were extracted.The most commonly used stimulation parameters in electroacupuncture treatment of CSR were 2 Hz of continuous wave,2/100 Hz of dilatational wave,intensity tolerance,30 min/time,1 time/day,10 times/course,and a total of 20 treatments.The top 3 most commonly used acupoints were Jingjiaji,Houxi(SI3)and Fengchi(GB20);the meridians of acupoints were mainly the large intestine meridian,small intestine meridian and gallbladder meridian;the core prescription of association rule analysis was"Jingjiaji,Houxi,Quchi,Fengchi,Jianjing,Waiguan,Tianzhu,Hegu,Dazhui",and the acupoint combination with the highest support was"Jingjiaji-Houxi".5 clusters were obtained through clustering analysis.Conclusion In electroacupuncture treatment for CSR,the selection of acupoints is primarily aimed at unblocking meridians to relieve pain and nourishing the liver and kidney.The core acupoints group is Jingjiaji-Houxi;the stimulation parameters demonstrate a certain degree of regularity and clustering,primarily consisting of low-frequency continuous wave and alternating density wave of low and high frequencies.