1.Optimizing extraction of microbial DNA from urine: Advancing urinary microbiome research in bladder cancer
Chuang-Ming ZHENG ; Ho Won KANG ; Seongmin MOON ; Young Joon BYUN ; Won Tae KIM ; Yung Hyun CHOI ; Sung-Kwon MOON ; Xuan-Mei PIAO ; Seok Joong YUN
Investigative and Clinical Urology 2025;66(3):272-280
Purpose:
This study aimed to evaluate and optimize microbial DNA extraction methods from urine, a non-invasive sample source, to enhance DNA quality, purity, and reliability for urinary microbiome research and biomarker discovery in bladder cancer.
Materials and Methods:
A total of 302 individuals (258 with genitourinary cancers and 44 with benign urologic diseases) participated in this study. Urine samples were collected via sterile catheterization, resulting in 445 vials for microbial analysis. DNA extraction was performed using three protocols: the standard protocol (SP), water dilution protocol (WDP), and chelation-assisted protocol (CAP). DNA quality (concentration, purity, and contamination levels) was assessed using NanoDrop spectrophotometry.Microbial analysis was conducted on 138 samples (108 cancerous and 30 benign) using 16S rRNA sequencing. Prior to sequencing on the Illumina MiSeq platform, Victor 3 fluorometry was used for validation.
Results:
WDP outperformed other methods, achieving significantly higher 260/280 and 260/230 ratios, indicating superior DNA purity and reduced contamination, while maintaining reliable DNA yields. CAP was excluded due to poor performance across all metrics. Microbial abundance was significantly higher in WDP-extracted samples (p<0.0001), whereas SP demonstrated higher alpha diversity indices (p<0.01), likely due to improved detection of low-abundance taxa. Beta diversity analysis showed no significant compositional differences between SP and WDP (p=1.0), supporting the reliability of WDP for microbiome research.
Conclusions
WDP is a highly effective and reliable method for microbial DNA extraction from urine, ensuring high-quality and reproducible results. Future research should address sample variability and crystal precipitation to further refine microbiome-based diagnostics and therapeutics.
2.Pathogenesis and treatment of "inflammation cancer transformation" of ulcerative colitis based on "Kenang" theory.
Jia-Kang XIE ; Xiao-Ning XU ; Feng-Ting AI ; Shao-Xi LI ; Yun AN ; Xuan GONG ; Yong CAO
China Journal of Chinese Materia Medica 2025;50(8):2298-2304
Ulcerative colitis(UC) is a recurrent, chronic, nonspecific inflammatory bowel disease. The longer the course of the disease, the higher the risk of cancerization. In recent years, the incidence and mortality rates of colon cancer in China have been increasing year by year, seriously threatening the life and health of patients. Therefore, studying the mechanism of "inflammation cancer transformation" in UC and conducting early intervention is crucial. The "Kenang" theory is an important component of traditional Chinese medicine(TCM) theory of phlegm and blood stasis. It is based on the coexistence of phlegm and blood stasis in the body and deeply explores the pathogenic syndromes and characteristics of phlegm and blood stasis. Kenang is a pathological product formed when long-term Qi stagnation leads to the internal formation of phlegm and blood stasis, which is hidden deep within the body. It is characterized by being hidden, progressive, and difficult to treat. The etiology and pathogenesis of "inflammation cancer transformation" in UC are consistent with the connotation of the "Kenang" theory. The internal condition for the development of UC "inflammation cancer transformation" is the deficiency of healthy Qi, with Qi stagnation being the key pathological mechanism. Phlegm and blood stasis are the main pathogenic factors. Phlegm and blood stasis accumulate in the body over time and can produce cancer toxins. Due to the depletion of healthy Qi and a weakened constitution, the body is unable to limit the proliferation and invasion of cancer toxins, eventually leading to cancer transformation in UC. In clinical treatment, the focus should be on removing phlegm and blood stasis, with syndrome differentiation and treatment based on three basic principles: supporting healthy Qi to strengthen the body's foundation, resolving phlegm and blood stasis to break up the Kenang, and regulating Qi and blood to smooth the flow of energy and resolve stagnation. This approach helps to dismantle the Kenang, delay, block, or even reverse the cancerization process of UC, reduce the risk of "inflammation cancer transformation", improve the patient's quality of life, and provide new perspectives and strategies for early intervention in the development of colon cancer.
Humans
;
Colitis, Ulcerative/immunology*
;
Medicine, Chinese Traditional
;
Drugs, Chinese Herbal/therapeutic use*
;
Cell Transformation, Neoplastic
3.Rutaecarpine Attenuates Monosodium Urate Crystal-Induced Gouty Inflammation via Inhibition of TNFR-MAPK/NF-κB and NLRP3 Inflammasome Signaling Pathways.
Min LI ; Zhu-Jun YIN ; Li LI ; Yun-Yun QUAN ; Ting WANG ; Xin ZHU ; Rui-Rong TAN ; Jin ZENG ; Hua HUA ; Qin-Xuan WU ; Jun-Ning ZHAO
Chinese journal of integrative medicine 2025;31(7):590-599
OBJECTIVE:
To investigate the anti-inflammatory effect of rutaecarpine (RUT) on monosodium urate crystal (MSU)-induced murine peritonitis in mice and further explored the underlying mechanism of RUT in lipopolysaccharide (LPS)/MSU-induced gout model in vitro.
METHODS:
In MSU-induced mice, 36 male C57BL/6 mice were randomly divided into 6 groups of 8 mice each group, including the control group, model group, RUT low-, medium-, and high-doses groups, and prednisone acetate group. The mice in each group were orally administered the corresponding drugs or vehicle once a day for 7 consecutive days. The gout inflammation model was established by intraperitoneal injection of MSU to evaluate the anti-gout inflammatory effects of RUT. Then the proinflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and the proportions of infiltrating neutrophils cytokines were detected by flow cytometry. In LPS/MSU-treated or untreated THP-1 macrophages, cell viability was observed by cell counting kit 8 and proinflammatory cytokines were measured by ELISA. The percentage of pyroptotic cells were detected by flow cytometry. Respectively, the mRNA and protein levels were measured by real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot, the nuclear translocation of nuclear factor κB (NF-κB) p65 was observed by laser confocal imaging. Additionally, surface plasmon resonance (SPR) and molecular docking were applied to validate the binding ability of RUT components to tumor necrosis factor α (TNF-α) targets.
RESULTS:
RUT reduced the levels of infiltrating neutrophils and monocytes and decreased the levels of the proinflammatory cytokines interleukin 1β (IL-1β) and interleukin 6 (IL-6, all P<0.01). In vitro, RUT reduced the production of IL-1β, IL-6 and TNF-α. In addition, RT-PCR revealed the inhibitory effects of RUT on the mRNA levels of IL-1β, IL-6, cyclooxygenase-2 and TNF-α (P<0.05 or P<0.01). Mechanistically, RUT markedly reduced protein expressions of tumor necrosis factor receptor (TNFR), phospho-mitogen-activated protein kinase (p-MAPK), phospho-extracellular signal-regulated kinase, phospho-c-Jun N-terminal kinase, phospho-NF-κB, phospho-kinase α/β, NOD-like receptor thermal protein domain associated protein 3 (NLRPS), cleaved-cysteinyl aspartate specific proteinase-1 and cleaved-gasdermin D in macrophages (P<0.05 or P<0.01). Molecularly, SPR revealed that RUT bound to TNF-α with a calculated equilibrium dissociation constant of 31.7 µmol/L. Molecular docking further confirmed that RUT could interact directly with the TNF-α protein via hydrogen bonding, van der Waals interactions, and carbon-hydrogen bonding.
CONCLUSION
RUT alleviated MSU-induced peritonitis and inhibited the TNFR1-MAPK/NF-κB and NLRP3 inflammasome signaling pathway to attenuate gouty inflammation induced by LPS/MSU in THP-1 macrophages, suggesting that RUT could be a potential therapeutic candidate for gout.
Animals
;
NF-kappa B/metabolism*
;
Male
;
Indole Alkaloids/therapeutic use*
;
Signal Transduction/drug effects*
;
Mice, Inbred C57BL
;
Inflammation/complications*
;
Uric Acid
;
Quinazolines/therapeutic use*
;
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*
;
Humans
;
Gout/chemically induced*
;
Inflammasomes/metabolism*
;
Cytokines/metabolism*
;
THP-1 Cells
;
Mitogen-Activated Protein Kinases/metabolism*
;
Mice
;
Molecular Docking Simulation
;
Lipopolysaccharides
;
Quinazolinones
4.Predicting Postoperative Circulatory Complications in Older Patients: A Machine Learning Approach.
Xiao Yun HU ; Wei Xuan SHENG ; Kang YU ; Jie Tai DUO ; Peng Fei LIU ; Ya Wei LI ; Dong Xin WANG ; Hui Hui MIAO
Biomedical and Environmental Sciences 2025;38(3):328-340
OBJECTIVE:
This study examines utilizes the advantages of machine learning algorithms to discern key determinants in prognosticate postoperative circulatory complications (PCCs) for older patients.
METHODS:
This secondary analysis of data from a randomized controlled trial involved 1,720 elderly participants in five tertiary hospitals in Beijing, China. Participants aged 60-90 years undergoing major non-cardiac surgery under general anesthesia. The primary outcome metric of the study was the occurrence of PCCs, according to the European Society of Cardiology and the European Society of Anaesthesiology diagnostic criteria. The analysis metrics contained 67 candidate variables, including baseline characteristics, laboratory tests, and scale assessments.
RESULTS:
Our feature selection process identified key variables that significantly impact patient outcomes, including the duration of ICU stay, surgery, and anesthesia; APACHE-II score; intraoperative average heart rate and blood loss; cumulative opioid use during surgery; patient age; VAS-Move-Median score on the 1st to 3rd day; Charlson comorbidity score; volumes of intraoperative plasma, crystalloid, and colloid fluids; cumulative red blood cell transfusion during surgery; and endotracheal intubation duration. Notably, our Random Forest model demonstrated exceptional performance with an accuracy of 0.9872.
CONCLUSION
We have developed and validated an algorithm for predicting PCCs in elderly patients by identifying key risk factors.
Aged
;
Aged, 80 and over
;
Female
;
Humans
;
Male
;
Middle Aged
;
Cardiovascular Diseases/etiology*
;
Machine Learning
;
Postoperative Complications/etiology*
;
Risk Factors
;
Randomized Controlled Trials as Topic
;
Secondary Data Analysis
5.Changes in Esophageal Cancer Survival: A Global Review of Survival Analysis from Cancer Registration Data over the Past Three Decades.
Zhuo Jun YE ; Dan Ni YANG ; Yu JIANG ; Yu Xuan XIAO ; Zhuo Ying LI ; Yu Ting TAN ; Hui Yun YUAN ; Yong Bing XIANG
Biomedical and Environmental Sciences 2025;38(5):571-584
OBJECTIVE:
To describe survival trends and global patterns of esophageal cancer (EC) using survival data from population-based cancer registries.
METHODS:
We systematically searched PubMed, EMBASE, Web of Science, SEER, and SinoMed databases for articles published up to 31 December 2023. Eligible EC survival estimates were evaluated according to country or region, period, sex, age group, pathology, and disease stage.
RESULTS:
After 2010, Jordan exhibited the highest age-standardized 5-year relative survival rates (RSRs)/net survival rates (NSRs) at 41.1% between 2010 and 2014, while India had the lowest, at 4.1%. Survival rates generally improved with diagnostic age across most countries, with significant increases in South Korea and China, of 12.7% and 10.5% between 2000 and 2017, respectively. Survival was higher among women compared to men, ranging from 0.4%-10.9%. Survival rates for adenocarcinoma and squamous cell carcinoma were similar, differing by about 4%. In China, the highest age-standardized RSRs/NSRs was 33.4% between 2015 and 2017. Meanwhile, the lowest was 5.3%, in Qidong (Jiangsu province) between 1992-1996.
CONCLUSION
Global EC survival rates have improved significantly in recent decades, but substantial geographical, sex, and age disparities still exist. In Asia, squamous cell carcinoma demonstrated superior survival rates compared to adenocarcinoma, while the opposite trend was observed in Western countries. Future research should clarify the prognostic factors influencing EC survival and tailor prevention and screening strategies to the changing EC survival patterns.
Humans
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Esophageal Neoplasms/mortality*
;
Registries
;
Male
;
Female
;
Survival Analysis
;
Middle Aged
;
Survival Rate
;
Aged
;
Global Health
6.Development of low-oxygen mixed gas generator for pilot hypoxia testing
Lin-Xia LI ; Jia-Ling XU ; Guo-Yun MAO ; Yao-Xuan JI ; Jin MA ; Yun-Ying WANG
Chinese Medical Equipment Journal 2024;45(7):24-28
Objective To develop a low-oxygen mixed gas generator to make up for the deficiencies of low-pressure chambers and load-resistant hypoxia trainers during pilot hypoxia tolerance testing.Methods The device prepared the low-oxygen gas with the principle of gas separation,which was composed of a Sunsource OLF1100D-220AF air compressor,a SMC IDG75SAM4-03 filter,a buffer tank,an AIR Products PA3010 integrated assembly,a control box,sensors and regulators.The sensors included the pressure sensor,flow sensor,concentration sensor and dew point temperature sensor,and the control box consisted of a main control board,a power supply module,a transmission module,a communication module,a display and a housing.The embedded control software of the device was developed with KEIL 5 and C++.Results The device developed prepared the low-oxygen gas with the volume fraction being 4%to 18%and the maximum error of volume fraction being 0.05%,and the main components of the prepared gas met the technical requirements of medical oxygen as stipulated in GB 8982-2009 Oxygen supplies for medicine and aircraft breathing.Conclusion The low-oxygen gas prepared by the device has its volume fraction precisely controlled and can be used for hypoxia tolerance testing and acclimation training for pilots.[Chinese Medical Equipment Journal,2024,45(7):24-28]
7.COL6A1 expression as a potential prognostic biomarker for risk stratification of T1 high grade bladder cancer: Unveiling the aggressive nature of a distinct non-muscle invasive subtype
Kyeong KIM ; Young Joon BYUN ; Chuang-Ming ZHENG ; Sungmin MOON ; Soo Jeong JO ; Ho Won KANG ; Won Tae KIM ; Yung Hyun CHOI ; Sung-Kwon MOON ; Wun-Jae KIM ; Xuan-Mei PIAO ; Seok Joong YUN
Investigative and Clinical Urology 2024;65(1):94-103
Purpose:
T1 high grade (T1HG) bladder cancer (BC) is a type of non-muscle invasive BC (NMIBC) that is recognized as an aggressive subtype with a heightened propensity for progression. Current risk stratification methods for NMIBC rely on clinicopathological indicators; however, these approaches do not adequately capture the aggressive nature of T1HG BC. Thus, new, more accurate biomarkers for T1HG risk stratification are needed. Here, we enrolled three different patient cohorts and investigated expression of collagen type VI alpha 1 (COL6A1), a key component of the extracellular matrix, at different stages and grades of BC, with a specific focus on T1HG BC.
Materials and Methods:
Samples from 298 BC patients were subjected to RNA sequencing and real-time polymerase chain reaction.
Results:
We found that T1HG BC and muscle invasive BC (MIBC) exhibited comparable expression of COL6A1, which was significantly higher than that by other NMIBC subtypes. In particular, T1HG patients who later progressed to MIBC had considerably higher expression of COL6A1 than Ta, T1 low grade patients, and patients that did not progress, highlighting the aggressive nature and higher risk of progression associated with T1HG BC. Moreover, Cox and Kaplan–Meier survival analyses revealed a significant association between elevated expression of COL6A1 and poor progression-free survival of T1HG BC patients (multivariate Cox hazard ratio, 16.812; 95% confidence interval, 3.283–86.095; p=0.001 and p=0.0002 [log-rank test]).
Conclusions
These findings suggest that COL6A1 may be a promising biomarker for risk stratification of T1HG BC, offering valuable insight into disease prognosis and guidance of personalized treatment decisions.
8.Application of group guidance based on intelligent communication matrix in patients with type 2 diabetes mellitus
Cuixiang XUAN ; Yan HU ; Lanlan ZHANG ; Fang LI ; Guangfeng TANG ; Daoqin ZHANG ; Yun CHEN ; Shiguang LI
Chinese Journal of Modern Nursing 2024;30(3):384-388
Objective:To explore the application effect of group guidance based on intelligent communication matrix in patients with type 2 diabetes mellitus (T2DM) .Methods:Using the convenient sampling method, a total of 60 T2DM patients with poor self-management ability who scored less than 19.6 on Summary of Diabetes Self Care Activities (SDSCA) in the Affiliated Chuzhou Hospital of Anhui Medical University were selected as the research objects from May to June 2020. They were randomly divide into the observation group ( n=30) and the control group ( n=30). The observation group was given continuous group guidance based on intelligent communication matrix, while the control group was given routine health guidance. The SDSCA score, body mass index and glycosylated hemoglobin (HbA1c) before and after intervention were compared between the two groups. Results:After intervention, the SDSCA score of the observation group was higher than that of the control group, HbA1c was lower than that of the control group and body mass index was lower than that of the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The group guidance based on intelligent communication matrix can improve the self-management level of T2DM patients and reduce their body mass index and HbA1c levels, which has clinical application value.
9.Study on improvement effect of Danggui Shaoyao San on edema in rats with nephrotic syndrome and its mechanism
Xiao-Wen MA ; Sheng-Nan FAN ; Zai-Ping XU ; Qing-Zhen XIANG ; Zi-Hua XUAN ; Yun-Lai WANG ; Fan XU
Chinese Pharmacological Bulletin 2024;40(6):1172-1178
Aim To explore the effect of Danggui Shaoyao San on edema in rats with nephrotic syndrome and the underlying mechanism.Methods Rats were randomly divided into control group,model group,Danggui Shaoyao San group(17.2 g·kg-1·d-1),losartan group(30 mg·kg-1·d-1)and tolvaptan group(3 mg·kg-1·d-1).The rat model of nephrot-ic syndrome was established by tail vein injection of adriamycin.After four weeks of treatment,the levels of renal function and 24 h urinary protein were detected.The distribution of aquaporin 2(AQP2)and pS256-AQP2 in renal tissue was detected by immunohisto-chemistry.The levels of plasma arginine vasopressin(AVP)and angiotensin Ⅱ(Ang Ⅱ)were measured by radioimmunoassay.The expressions of renal AQP2,pS256-AQP2,angiotensin type 1 receptor(AT1R),arginine vasopressin receptor 2(V2R)protein and mRNA were measured by Western blot and RT-PCR,respectively.Results The three drugs could improve renal function,reduce proteinuria,decrease plasma AVP and Ang Ⅱ levels,and down-regulate AQP2 and pS256-AQP2 protein and mRNA expression in model rats.Danggui Shaoyao San and tolvaptan were more ef-fective than losartan in reducing plasma AVP levels.Conclusions Danggui Shaoyao San may regulate the expression of AQP2 by reducing the levels of AVP and Ang Ⅱ,and improve the edema of nephrotic syndrome rats.
10.Preliminary study on delaying aging induced thymus degeneration in SAMP6 mice with Bazi Bushen capsule
Zhao-Dong LI ; Yin-Xiao CHEN ; Bo-Yang GONG ; Zhe XU ; Zhi-Xian YU ; Yue-Xuan SHI ; Yan-Fei PENG ; Yu-Hong BIAN ; Yun-Long HOU ; Xiang-Ling WANG ; Shu-Wu ZHAO
Chinese Pharmacological Bulletin 2024;40(6):1186-1192
Aim To explore the improvement effect of Bazi Bushen capsule on thymic degeneration in SAMP6 mice and the possible mechanism.Methods Twenty 12 week old male SAMP6 mice were randomly divided into the model group(SAMP6)and the Bazi Busheng capsule treatment group(SAMP6+BZBS).Ten SAMR1 mice were assigned to a homologous control group(SAMR1).The SAMP6+BZBS group was oral-ly administered Bazi Bushen capsule suspension(2.8 g·kg-1)daily,while the other two groups were orally administered an equal amount of distilled water.After nine weeks of administration,the morphology of the thymus in each group was observed and the thymus in-dex was calculated;HE staining was used to observe the structural changes of thymus tissue;SA-β-gal stai-ning was used to detect thymic aging;flow cytometry was used to detect the proportion of thymic CD3+T cells in each group;Western blot was used to detect the levels of p16,Bax,Bcl-2,and cleaved caspase-3 proteins in thymus;immunofluorescence was applied to detect the proportion of cortical thymic epithelial cells in each group;ELISA was employed to detect IL-7 lev-els in thymus.Results Compared with the SAMP6 group,the thymic index of the SAMP6+BZBS group significantly increased(P<0.05);the disordered thy-mic structure was significantly improved;the positive proportion of SA-β-gal staining significantly decreased(P<0.01);the proportion of CD3+T cells apparently increased(P<0.05);the level of p16 protein signifi-cantly decreased(P<0.05);the level of Bcl-2 pro-tein significantly increased(P<0.05),while the lev-el of cleaved caspase-3 protein markedly decreased(P<0.05);the proportion of cortical thymic epithelial cells evidently increased;the level of IL-7 significantly increased(P<0.01).Conclusions Bazi Bushen capsule can delay thymic degeneration,inhibit cell ap-optosis in thymus and promote thymic cell development in SAMP6 mice,which may be related to increasing the proportion of cortical thymic epithelial cells and promoting IL-7 secretion.

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