1.Effect of Oral Sodium Butyrate on Skeletal Muscle Atrophy via The Gut-muscle Axis in Antibiotic-pretreated CT26 Tumor-bearing Mice and Its Mechanism
Shu-Ling ZHANG ; Jun-Wei WANG ; Shi-Liang HU ; Tu-Tu WANG ; Shun-Chang LI ; Jia FAN ; Jun-Zhi SUN
Progress in Biochemistry and Biophysics 2026;53(3):724-739
ObjectiveTo explore the effect of oral sodium butyrate on skeletal muscle atrophy in CT26 tumor mice through the gut microbiota-skeletal muscle axis and its potential mechanism. MethodsSixty SPF BALB/c male mice aged 8 weeks were randomly divided into a normal control group (NC, n=18) and a ABX-depleted group (ABX, n=42). The ABX mice were pretreated with a quadruple antibiotic cocktail via oral gavage (0.2 ml per administration, once daily, 6 d per week, for 2 weeks), whereas NC received an equal volume of sterile water. The quadruple antibiotic cocktail consisted of metronidazole (1 g/L), vancomycin (0.5 g/L), ampicillin (1 g/L), and gentamicin (1 g/L). Following successful pretreatment, six mice from each group were randomly selected for gut microbiota sequencing analysis and designated as the Abx group and the NC0 group, respectively. Theremaining mice in ABX were subcutaneously inoculated in the dorsum with 0.2 ml of CT26 cell suspension (at a cell density of 1×107/ml). Then these mice were randomly allocated into three subgroups: a control tumor bearing model group (0_NaB, n=12), a tumor-bearing model group receiving low-dose oral sodium butyrate (L_NaB, n=12), a tumor-bearing model group receiving high-dose oral sodium butyrate (H_NaB, n=12). And mice in NC were inoculated at the same site with 0.2 ml of normal saline. The administration dose for L_NaB was 0.3 g/(kg·d), that for H_NaB was 0.5 g/(kg·d), while NC and 0_NaB were given the same volume of normal saline (0.2ml per time, once daily, 6 d per week, for 4 weeks). The general condition of mice was monitored, and forelimb grip strength gastrocnemius muscle mass and its muscle fiber cross-sectional area were measured for each group. The structural changes in gut microbiota were assessed by 16S rRNA sequencing of cecal contents. Pathological alterations in the intestinal wall were examined via HE staining. Serum and gastrocnemius muscle levels of TNF‑α, IL-6, IL-1β, and LPS were quantified using ELISA. The protein expression of ZO-1 and occludin in the small intestine, as well as proteins associated with the TLR4/MyD88/NF-κB signaling pathway in the gastrocnemius muscle, were detected by Western blot analysis. Results(1) The alpha-diversity in Abx was significantly lower than that in NC0 (P<0.01), a significant decrease of the mass and muscle fiber cross-sectional area of the gastrocnemius (P<0.01), with the majority of gut microbiota being effectively depleted. (2) Compared with NC, the subcutaneous tumors of mice in 0_NaB were prominent, a significant increase of the mass and muscle fiber cross-sectional area of the gastrocnemius, accompanied by a significant decrease in body weight at the end of the 3th and 4th week (P<0.05), and a significant weakening of the forelimb grasping strength at the 5th and 6th week (P<0.01). Compared with 0_NaB, the tumor mass of mice in L_NaB and H_NaB showed a significant decreasing trend, and the grip strength of the forelimbs significantly increased at the 5th and 6th week (P<0.05, P<0.01). (3) Compared with 0_NaB, the Shannon and Observed species indices in α diversity of L_NaB and H_NaB were significantly increased (P<0.05). At the genus level, compared with 0_NaB, L_NaB exhibited a significant decrease in the relative abundance of Parasutterella (P< 0.01), while H_NaB showed significant reductions in the relative abundances of both Escherichia-Shigella and Parasutterella (P < 0.01). (4) Compared with 0_NaB, the small intestinal tissue structure in L_NaB and H_NaB was more intact, the infiltration of inflammatory cells was significantly reduced, and the capillaries were slightly dilated. The expression levels of ZO-1 and occludin proteins in L_NaB were significantly increased (P<0.01). (5) The LPS concentration in the gastrocnemius muscle and the protein expression levels of TLR4, MyD88, p-IκBα, and p-NF‑κB p65 in L_NaB and H_NaB were significantly lower than those in 0_NaB (P<0.05). The serum TNF‑α concentration in H_NaB and TNF-α concentration in the gastrocnemius muscle of the L_NaB and H_NaB were significantly lower than those in 0_NaB (P<0.05, P<0.01, P<0.01). ConclusionOral administration of NaB can improve gut microbiota α diversity, adjusting its composition, improving intestinal mucosal barrier function, reducing the LPS-induced pro-inflammatory response, and delaying skeletal muscle atrophy. The underlying mechanism may involve down regulation of TLR4/MyD88/NF-κB signaling in skeletal muscle.
2.Standards for the Application of Hemodynamic Monitoring Technology in Critical Care
Hua ZHAO ; Hongmin ZHANG ; Xin DING ; Huan CHEN ; Jun DUAN ; Wei DU ; Bo TANG ; Yuankai ZHOU ; Dongkai LI ; Xinchen WANG ; Cui WANG ; Gaosheng ZHOU ; Xiaoting WANG
Medical Journal of Peking Union Medical College Hospital 2026;17(1):73-85
With the rapid advancement of hemodynamic indices and monitoring technologies, their classification methods and application processes have become increasingly complex. Currently, no unified standard hasbeen established, making it difficult to fully meet the clinical requirements for hemodynamic management. To assist in hemodynamic monitoring assessment and therapeutic decision-making in critically ill patients, the Critical Hemodynamic Therapy Collaborative Group, in conjunction with the Critical Ultrasound Study Group, has jointly developed the Standard for the Application of Hemodynamic Monitoring Techniques in Critical Care. The first part of this standard systematically categorizes hemodynamic indicators into flow indicators, pressure and its derivative indicators, and tissue perfusion indicators, while elaborating on the clinical application of each. The second part establishes a standardized clinical implementation pathway for hemodynamic monitoring. It proposes a tiered monitoring strategy-comprising basic, advanced, indication-specific, and special scenario monitoring-tailored to different clinical settings. It emphasizes the central role of critical care ultrasound across all levels of monitoring and establishes hemodynamic assessment standards for organs such as the brain, kidneys, and gastrointestinal tract. This standard aims to provide a unified framework for clinical practice, teaching, training, and research in critical care medicine, thereby promoting standardized development within the discipline.
3.The effect of body mass index and inferior pulmonary ligament division on the residual lung expansion after right upper lobectomy: A retrospective cohort study in a single center
Guang MU ; Wenhao ZHANG ; Hongchang WANG ; Yan GU ; Chenghao FU ; Wentao XUE ; Shiyuan XIE ; Tong WANG ; Ke WEI ; Yang XIA ; Liang CHEN ; Jun WANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(02):261-266
Objective To analyze the effect of releasing the lower pulmonary ligament on right residual lung expansion after right upper lobe resection under different body mass index (BMI) levels. Methods The clinical data of patients who underwent thoracoscopic right upper lobe resection in the First Affiliated Hospital with Nanjing Medical University from 2021 to 2022 were retrospectively analyzed. Patients were divided into a group A (17 kg/m2<BMI≤23 kg/m2), a group B (23 kg/m2<BMI≤29 kg/m2) and a group C (BMI>29 kg/m2) according to BMI. The presence of residual cavity was judged by chest X-ray at 7-10 days after operation, the degree of compensation change of the right main bronchus angle was measured, and the changes in lung volume were determined by CT three-dimensional reconstruction. Results A total of 157 patients who underwent thoracoscopic right upper lobe resection were included, including 71 males and 86 females, with an average age of (59.7±11.2) years. There were 50 patients in the group A, 75 patients in the group B, and 32 patients in the group C. In the group A, compared with those without releasing the lower pulmonary ligament, patients with releasing had a lower incidence of postoperative residual cavity (P=0.016), greater changes in bronchus angle (P<0.001), and smaller changes in lung volume (P<0.001). In the group B and C, there was no significant effect of releasing the lower pulmonary ligament on postoperative residual cavity, bronchus angle, and lung volume changes (P>0.05). Conclusion For patients with thin and long body shape and low BMI, releasing the lower pulmonary ligament is helpful to promote the expansion of the residual lung after right upper lobe resection and reduce the occurrence of postoperative residual cavity in patients.
4.Clinical Efficacy of Modified Linggui Zhugan Tang in Patients with Obstructive Sleep Apnea-Hypopnea Syndrome of Spleen Deficiency and Dampness Obstruction with Blood Stasis Type and Its Effect on MIF, miR-223, and IL-18
Jun ZHANG ; Mengmei WEI ; Bo LI ; Yi YANG ; Changhui LINGHU ; Mingchang ZHANG ; Zhengxing GE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):171-179
ObjectiveTo investigate the intervention effects of modified Linggui Zhugan Tang (LGZGT) on patients with obstructive sleep apnea-hypopnea syndrome (OSAHS) of the spleen deficiency and dampness obstruction with blood stasis type, reveal its possible mechanisms, and provide a theoretical basis for the clinical treatment of OSAHS with traditional Chinese medicine (TCM). MethodsEighty OSAHS patients with spleen deficiency and dampness obstruction with blood stasis were randomly assigned to a control group and an observation group (1∶1) using a random number table, with 40 patients in each group. The control group received standard basic treatment combined with oral Doxofylline tablets, while the observation group received standard basic treatment combined with modified LGZGT. Serum levels of macrophage migration inhibitory factor (MIF), microRNA-223 (miR-223), and interleukin-18 (IL-18) were measured by enzyme-linked immunosorbent assay (ELISA), and the mRNA expression levels of MIF, miR-223, and IL-18 were measured by real-time quantitative polymerase chain reaction (Real-time PCR). After two months of treatment, the total clinical efficacy, apnea-hypopnea index (AHI), lowest nocturnal oxygen saturation (LSpO2), body mass index (BMI), TCM syndrome scores, and expression levels of MIF, miR-223, and IL-18 before and after treatment were compared between the two groups. Correlations between MIF, miR-223, IL-18 and AHI and LSpO2 were also analyzed. ResultsCompared with the control group, the observation group showed a significantly higher total clinical effective rate (P<0.01, Z=-3.49). Within the control group, no significant changes were observed in AHI, LSpO2, BMI, TCM syndrome scores, or MIF, miR-223, IL-18 levels and their mRNAs after treatment. In the observation group, AHI, BMI, TCM syndrome scores, and MIF and IL-18 levels and their mRNAs decreased significantly, while LSpO2 increased significantly (P<0.01). After treatment, compared with the control group, the observation group exhibited significantly lower AHI, BMI, TCM syndrome scores, and MIF and IL-18 levels and their mRNAs, and significantly higher LSpO2 (P<0.01). Correlation analysis showed that MIF and IL-18 were positively correlated with AHI (P<0.01) and negatively correlated with LSpO2 (P<0.01), whereas miR-223 was negatively correlated with AHI (P<0.01) and positively correlated with LSpO2 (P<0.01). ConclusionModified LGZGT may improve OSAHS of the spleen deficiency and dampness obstruction with blood stasis type by reducing airway inflammatory factors, alleviating airway inflammation, relieving airway edema and stenosis, and improving airway obstruction.
5.A Case of Tuberous Sclerosis Complex with Multiple Organ Involvement Caused by TSC2 Gene Mutation
Hongli ZHANG ; Jiayuan DAI ; Yan WANG ; Weihong ZHANG ; Wenbin MA ; Hanhui FU ; Chunxia HE ; Jun ZHENG ; Wenda WANG ; Wei ZUO ; Yaping LIU ; Min SHEN
JOURNAL OF RARE DISEASES 2026;5(1):60-67
Tuberous sclerosis complex (TSC) is an autosomal dominant genetic disorder primarily caused by pathogenic variants in the
6.Expert consensus on neoadjuvant PD-1 inhibitors for locally advanced oral squamous cell carcinoma (2026)
LI Jinsong ; LIAO Guiqing ; LI Longjiang ; ZHANG Chenping ; SHANG Chenping ; ZHANG Jie ; ZHONG Laiping ; LIU Bing ; CHEN Gang ; WEI Jianhua ; JI Tong ; LI Chunjie ; LIN Lisong ; REN Guoxin ; LI Yi ; SHANG Wei ; HAN Bing ; JIANG Canhua ; ZHANG Sheng ; SONG Ming ; LIU Xuekui ; WANG Anxun ; LIU Shuguang ; CHEN Zhanhong ; WANG Youyuan ; LIN Zhaoyu ; LI Haigang ; DUAN Xiaohui ; YE Ling ; ZHENG Jun ; WANG Jun ; LV Xiaozhi ; ZHU Lijun ; CAO Haotian
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(2):105-118
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.
7.Statistical analysis of vector monitoring in port areas of Shandong Province from 2017 to 2023
Huan-mei HAN ; Tao ZHANG ; Zhi-ping SU ; Rong-jun YAN ; Wei HUANG ; Wen-wen ZHANG
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):121-127
Objective To provide a scientific basis for vector prevention and control in port areas, vector monitoring has been conducted continuously to characterize the composition, density, and seasonal fluctuations of vectors across 21 port-areas of Shandong province from 2017 to 2023. Methods In accordance with the Regional Vector Monitoring Plan for Frontier Ports, vector surveillance of mosquitoes, flies, cockroaches, rodents, and other disease vectors was performed. Data were analyzed to describe vector community composition and identify seasonal fluctuations. Results A total of 601 rodents belonging to 1 family, 4 genera, and 4 species were captured. Three dominant rodent species inhabit port areas. In addition, 88 104 mosquitoes were captured and classified into 3 subfamilies, 4 genera, and 12 species; 38 959 flies were trapped and classified into 8 families,31 genera, and 51 species, including 9 dominant fly species in port-areas; and 54 672 captured cockroaches were classified into 2 families,4 genera, and 4 species, with Blattella germanica as the dominant species. Conclusions Significant vector community changes(P<0.005)have been identified after the epidemic. To improve the pertinence and efficacy of port health quarantine work, continuous and strong attention should be paid to vector compositions and population densities at Shandong ports.
8.Analysis of prediction of incidence and epidemiological characteristics of hand, foot and mouth disease among children aged 0-9 years in Guangzhou, 2013-2025
Chinese Journal of School Health 2026;47(7):1026-1030
Objective:
To analyze the epidemiological characteristics of hand, foot and mouth disease (HFMD) among children aged 0-9 years in Guangzhou, so as to provide reference basis for further exploring scientifically effective prevention and control measures.
Methods:
Reported cases of HFMD in children aged 0-9 years in Guangzhou City from 2013-2025 were selected from China disease prevention and control information system, and the incidence in different years was analyzed descriptively; the trends of the composition ratio and incidence in various age groups were analyzed by using the Joinpoint regression model, spatial autocorrelation analyses were performed at the district level, and the seasonal autoregressive integrated moving average model (SARIMA) was applied to predict annual incidence rate for 2026.
Results:
From 2013 to 2025, the average annual reported incidence rate of HFMD among children aged 0-9 years in Guangzhou was 2 513.98/100 000, showing a fluctuating trend. Peaks of incidence occurred from May to July and from September to October. The male to female ratio was 1.49∶1. The average annual incidence rate was 4 743.99/100 000 in the 0-1 years age group, 3 298.19/100 000 in the 2-5 years age group, and 544.22/ 100 000 in the 6-9 years age group. Joinpoint regression analysis showed a decreasing trend in the proportion of the 0-1 years age group (APC= -6.02 %, t =-3.77) and an increasing trend in the proportion of the 6-9 years age group (APC=16.39%, t =5.67) (both P < 0.05 ). No significant global spatial clustering was observed at the district level; however, local scattered clusters were identified in certain areas. Using the SARIMA(2,0,0)(2,0,0) 12 model, the predicted annual incidence rate of HFMD among children aged 0-9 years in Guangzhou in 2026 was 2 680.56/100 000.
Conclusions
From 2013 to 2025, the annual incidence rate of HFMD among children aged 0-9 years in Guangzhou exhibited a fluctuating trend; the highest incidence rate is observed in the 0-1 years age group, while the proportion of cases in the 6-9 years age group increase year by year;and local spatial clusters of abnormal distribution are identified. It is essential to enhance risk monitoring and health education in key populations and areas to mitigate the risk of infection in children.
9.Epidemiological characteristics of imported malaria cases in Guangxi Zhuang Autonomous Region from 2017 to 2024
Kangming LIN ; Shujiao WEI ; Jun LI ; Weiwei ZHANG ; Hui YAN ; Jun MENG
Chinese Journal of Schistosomiasis Control 2026;38(4):389-398
Objective To analyze the epidemiological characteristics of imported malaria cases in Guangxi Zhuang Autonomous Region from 2017 to 2024, so as to formulate the targeted control strategies for imported malaria. Methods The demographic and epidemiological data of malaria cases reported in Guangxi Zhuang Autonomous Region from 2017 to 2024 were collected through the China Disease Prevention and Control Information System. The source of malaria cases acquiring infections, composition of malaria parasite species, regional distribution of malaria cases, population characteristics of malaria cases, temporal distribution of malaria cases, time of diagnosis, deaths and institutions reporting malaria cases were statistically analyzed with a descriptive statistical method. Results A total of 1 308 malaria cases were reported in Guangxi Zhuang Autonomous Region from 2017 to 2024, and all cases were overseas imported cases, with no secondary cases identified. Plasmodium falciparum malaria (57.11%, 747/1 308) and P. ovale malaria (27.68%, 362/1 308) were two most common forms of malaria, and three deaths occurred among P. falciparum malaria cases. The number of malaria cases appeared an overall tendency towards a decline from 2017 to 2022, followed by a rise after 2023. The malaria cases predominantly acquired infections from Africa (1 284 cases, accounting for 98.17%), with Ghana (432 cases, accounting for 33.03%) and Cameroon (145 cases, accounting for 11.09%) as two most frequent origin countries, and P. falciparum malaria (58.02%, 745/1 284) was the predominant form of malaria among African imported malaria cases; however, P. vivax malaria cases comprised 89.58% (86/96) of all malaria cases derived from Ethiopia. In terms of regional distribution, malaria cases were highly concentrated in Nanning City (1 075 cases, accounting for 82.19%), with the most prominent situated in Shanglin County (903 cases, accounting for 69.04%), and these cases were predominantly found among individuals at ages of 20 to 59 years (1 280 cases, accounting for 97.86%), males (1 265 cases, accounting for 96.71%), and gold/coal mining workers (1 042 cases, accounting for 79.66%). In terms of temporal distribution, malaria cases were reported throughout the year, with the peak incidence in April (146 cases, accounting for 11.16%). The median duration from onset of disease to initial diagnosis was 1 (interquartile range, 2) day, and the median duration from initial diagnosis to definitive diagnosis was 1 (interquartile range, 1) day. Malaria cases were predominantly reported by medical institutions (1 075 cases, accounting for 82.19%), and the proportion of malaria cases reported by medical institutions increased from 75.92% in 2017 to 96.27% in 2024, while the proportion of malaria cases reported by disease control and prevention institutions reduced from 24.08% in 2017 to 3.73% in 2024. There was a significant difference in the proportion of malaria cases reported by medical institutions and disease control and prevention institutions from 2017 to 2024 (both P < 0.001). The 3 dead imported P. falciparum malaria cases predominantly received initial diagnosis at townshipand lower-level institutions (66.67%, 2/3), with 1 (interquartile range, 2) day median duration from onset of disease to initial diagnosis, 2 (interquartile range, 4) days median duration from initial diagnosis to definitive diagnosis, and 1 (interquartile range, 19) day median duration from definitive diagnosis to death. Conclusions Imported P. falciparum malaria from Africa is predominant among malaria cases in Guangxi Zhuang Autonomous Region, and malaria cases are highly concentrated in specific regions and among specific occupational groups. It is recommended to strengthen inter-departmental collaboration, implement refined management among labors going to Africa, improve the integration of medical treatment and disease prevention, and enhance the diagnosis and treatment of severe malaria cases.
10.Establishment and Evaluation of Different Disease-Syndrome Combination Mice Models of Knee Osteoarthritis with Cold-Dampness Stagnation Syndrome
Enrui HU ; Yibao WEI ; Deren LIU ; Aobuliaisan MAIMAITITUXUN ; Licheng ZHANG ; Jiachen SONG ; Jun MAO ; Taiyang LIAO ; Peimin WANG
Journal of Traditional Chinese Medicine 2026;67(19):2120-2129
ObjectiveTo establish and evaluate different disease-syndrome combination mice models of knee osteoarthritis (KOA) with cold-dampness obstruction syndrome, and to identify the optimal modeling method. MethodsEighty mice were randomly divided into a control group, a treadmill plus climate chamber group, an anterior cruciate ligament transection (ACLT) plus climate chamber group, an ACLT group, a destabilization of the medial meniscus (DMM) plus climate chamber group, a DMM group, an intra-articular injection plus climate chamber group, and an intra-articular injection group, with 10 mice in each group. Mice in the ACLT group underwent surgical transection of the knee anterior cruciate ligament to establish the model, while those in the DMM underwent surgical transection of the medial meniscal tibial ligament of the knee, and those in the intra-articular injection group were modeled by an intra-articular injection of 50 μl of normal saline containing 1mg of monosodium iodoacetate (MIA) into the medial side of the patellar ligament. Mice in the ACLT plus climate chamber, DMM plus climate chamber, and intra-articular injection plus climate chamber groups were placed in an artificial climate chamber (relative humidity 60%, temperature 4 ℃, wind speed 6 m/s) simultaneously with aforementioned interventions, once daily for 2 h each time, for 14 consecutive days. Mice in the control group underwent sham surgery, involving only the incision of the skin and subcutaneous tissue at the knee joint to expose the articular cavity, followed by layer-by-layer suturing. Mice in the treadmill plus climate chamber group were placed on an animal treadmill starting on the 4th day after the sham operation, undergoing high-intensity treadmill exercise at a speed of 20 m/min, an inclination of 5°, for 30min per session, 5 times per week for 28 consecutive days, while being housed in the artificial climate chamber. Cold pain thresholds of the mice in each group were compared using the cold plate test and the acetone low-temperature test before and after modeling. Following modeling, knee joint pathological changes were observed using HE and Safranin O-Fast Green staining, and quantified using Mankin and Osteoarthritis Research Society International (OARSI) scores. Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured by ELISA. Protein and mRNA expression levels of pain-related markers, including calcitonin gene-related peptide (CGRP), protein gene product 9.5 (PGP9.5), and cellular proto-oncogene Fos (c-Fos), and the cold hyperalgesia-related markers including transient receptor potential ankyrin 1 (TRPA1) and transient receptor potential melastatin 8 (TRPM8) in the dorsal root ganglion (DRG) tissues were detected by Western Blot and qPCR. TRPA1 and TRPM8 fluorescent intensity in the DRG was detected by immunofluorescence assay. ResultsCompared to the control group, the remaining groups showed decreased cold pain thresholds in the cold plate tese, increased cold pain thresholds in the acetone low-temperature test, elevated Mankin and OARSI scores, and increased serum TNF-α and IL-1β levels, upregulated protein and mRNA expression levels of CGRP, PGP9.5, c-Fos, TRPA1, and TRPM8 in the DRG, and increased immunofluorescence intensities of TRPA1 and TRPM8 in the DRG tissue (P<0.05 or P<0.01), along with severe pathological damage and cartilage degeneration in the knee joints. Compared to the ACLT, DMM, and intra-articular injection groups, the ACLT plus climate chamber, DMM plus climate chamber, and intra-articular injection plus climate chamber groups exhibited more significant changes in all the aforementioned indicators (P<0.01). Furthermore, changes in the aforementioned indicators were more pronounced in the ACLT plus climate chamber group compared to the Treadmill plus climate chamber group, DMM plus climate chamber and intra-articular injection plus climate chamber group (P<0.01), exhibiting the most severe degree of pathological damage and cartilage degeneration. ConclusionThe combination of an artificial climate chamber can successfully establish a disease-syndrome combined animal model of KOA with cold-damp obstruction syndrome, with the ACLT combined with the artificial climate chamber being the optimal modeling approach.


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