1.The Effect of Qishao Tongbi Capsule (芪芍通痹胶囊) on the Wnt/β-catenin Pathway in a Rat Model of Intervertebral Disc Degeneration
Yumen XUE ; Xilin XU ; Wei HAN ; Jiaben XU ; Wenting XU ; Zelin LIU ; Xiaofeng ZHANG
Journal of Traditional Chinese Medicine 2025;66(1):79-88
ObjectiveTo explore the possible mechanism of Qishao Tongbi Capsule (芪芍通痹胶囊, QTC) in the treatment of intervertebral disc degeneration (IDD). MethodsSeventy-five rats were randomly divided into control group, model group, low-dose QTC group, high-dose QTC group, high-dose QTC +agonist group, with 15 rats in each group. Except for the control group, all other groups were subjected to a fibrous ring puncture to prepare an IDD model. After modeling, rats in low-dose QTC group and high-dose QTC group were given QTC at doses of 0.2 and 0.8 g/(kg·d) by gavage, respectively. Rats in high-dose QTC+ agonist group was given QTC at 0.8 g/(kg·d) and SKL2001 solution at 10 mg/(kg·d) by gavage. The control group and model group were given 10 ml/(kg·d) distilled water by gavage. All treatments were given once a day for 4 consecutive weeks. After treatment, X-ray and magnetic resonance imaging (MRI) were used to detect IDD degree. Hematoxylin-eosin (HE) staining and Safranin O-Fast Green staining were used to observe the morphological changes of the intervertebral disc tissue. Immunohistochemical staining was performed to examine the levels of proteoglycan, type Ⅱ collagen (COL Ⅱ), and matrix metalloproteinase-3 (MMP-3) in the intervertebral disc tissue. Western blotting was used to detect the extracellular matrix (ECM)-related proteins (proteoglycan, COL Ⅱ, MMP-3, MMP-9, MMP-13), aging-related proteins (P53, P21, P16), apoptosis related proteins, including B-cell lymphoma/leukemia 2 (BCL-2), BCL-2 related X protein (BAX), Cleaved Caspase-3, and Wnt/β-catenin pathway related proteins such as Wnt3a, glycogen synthase kinase-3β (GSK-3β) and β-catenin in the intervertebral disc nucleus pulposus (NP) tissue. Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) was used to assess the mRNA expression of Wnt3a, GSK-3β, and β-catenin in intervertebral disc tissue. ResultsCompared with the model group, rats in the low-dose QTC group and high-dose QTC group exhibited improved DHI, decreased Pfirmann grading, and alleviated IDD. The structural integrity of the NP and annulus fibrosus increased, and the number of the NP increased. The levels of proteoglycan, COL Ⅱ, BCL-2 and GSK-3β increased, while the levels of MMP-3, MMP-9, MMP-13, P53, P21, P16, BAX, Cleaved Caspase-3, Wnt3a and β-catenin protein decreased. The mRNA expression of Wnt3a and β-catenin mRNA decreased, while GSK-3β mRNA expression increased (P<0.05). Compared with the low-dose QTC group, the high-dose QTC group showed further improvements in DHI, decrease in Pfirrmann grading (P<0.05), and greater alleviation of IDD. The structural integrity of NP and annulus fibrosus was further enhanced, and the number of NP cells further increased. The levels of proteoglycan, COL Ⅱ, BCL-2 and GSK-3β were higher, while the levels of MMP-3, MMP-9, MMP-13, P53, P21, P16, BAX, Cleaved Caspase-3, Wnt3a and β-catenin were lower. The mRNA expression of Wnt3a and β-catenin decreased, while GSK-3β mRNA expression increased (P<0.05). Compared with the high-dose QTC group, the high-dose QTC +agonist group showed a decrease of DHI, an increase of Pfirmann grading (P<0.05), significant aggravation of IDD, reduction in structural integrity of the NP and annulus fibrosus, a decrease of NP cell count, lower levels of proteoglycan, COL Ⅱ, BCL-2 and GSK-3β, and higher levels of MMP-3, MMP-9, MMP-13, P53, P21, P16, BAX and Cleaved Caspase-3. Additionally, GSK-3β mRNA expression decreased (P<0.05). ConclusionQTC can inhibit NP cell aging, apoptosis, and ECM degradation in IDD rats, and its therapeutic effect may be mediated through the inhibition of the Wnt/β-catenin pathway.
2.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
3.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
4.Primary prostatic signet ring cell carcinoma:a report of 6 cases and literature review
Xiaofeng WANG ; Chengbiao CHU ; Xun WANG ; Tingzheng WANG ; Feifei ZHANG ; Wei CHEN ; Linfeng XU ; Qing ZHANG ; Hongqian GUO
Journal of Modern Urology 2025;30(4):290-295
Objective: To explore the diagnosis, treatment and prognosis of primary prostatic signet ring cell carcinoma (SRCC), so as to provide reference for the clinical diagnosis and treatment. Methods: A retrospective analysis was conducted on the clinical data of 6 patients with primary prostatic SRCC treated in Nanjing Drum Tower Hospital during Nov.2020 and Sep.2024.The clinical manifestations, imaging features, treatment methods, histological characteristics and prognosis were summarized. Results: The average age of the patients was (72.00±4.28) years.Varying degrees of dysuria occurred in 4 patients. All patients underwent multi-parametric magnetic resonance imaging (mpMRI) examination before surgery, and the results indicated typical prostate cancer.Preoperative biopsies showed high-grade (Gleason 8-10) prostate acinar adenocarcinoma.Postoperative pathological diagnoses were mixed types of prostate acinar adenocarcinoma and SRCC, and no metastasis was found in the pelvic lymph nodes.All patients were followed up for 1 to 46 months after surgery and are currently alive.Robot-assisted laparoscopic radical prostatectomy only was performed in 3 cases; apalutamide and leuprolide/triptorelin was administered after surgery in 2 cases; bicalutamide + goserelin was administered after surgery in 1 case, who developed bladder metastasis of prostate cancer 24 months later, and the serum prostate-specific antigen (PSA) concentration decreased to a safe level (<0.2 ng/mL) after the use of darolutamide with radiotherapy.No recurrence or metastasis was found in the remaining patients. Conclusion: Primary prostatic SRCC is a rare and highly aggressive malignant tumor of the prostate.The diagnosis depends on pathological examinations due to lack of specific imaging features and clinical manifestations.The prognosis is poor, and there is currently no standardized treatment.The combined use of surgery, hormonotherapy and radiotherapy can help improve the survival rate of patients.
5.RP11-789C1.1 inhibits gastric cancer cell proliferation and accelerates apoptosis via the ATR/CHK1 signaling pathway
Wenwei LIU ; Wei FENG ; Yongxin ZHANG ; Tianxiang LEI ; Xiaofeng WANG ; Tang QIAO ; Zehong CHEN ; Wu SONG
Chinese Medical Journal 2024;137(15):1835-1843
Background::Long non-coding RNAs (lncRNAs) plays an important role in the progression of gastric cancer (GC). Their involvement ranges from genetic regulation to cancer progression. However, the mechanistic roles of RP11-789C1.1 in GC are not fully understood.Methods::We identified the expression of lncRNA RP11-789C1.1 in GC tissues and cell lines by real-time fluorescent quantitative polymerase chain reaction. A series of functional experiments revealed the effect of RP11-789C1.1 on the proliferation of GC cells. In vivo experiments verified the effect of RP11-789C1.1 on the biological behavior of a GC cell line. RNA pull-down unveiled RP11-789C1.1 interacting proteins. Western blot analysis indicated the downstream pathway changes of RP11-789C1.1, and an oxaliplatin dosing experiment disclosed the influence of RP11-789C1.1 on the drug sensitivity of oxaliplatin. Results::Our results demonstrated that RP11-789C1.1 inhibited the proliferation of GC cells and promoted the apoptosis of GC cells. Mechanistically, RP11-789C1.1 inhibited checkpoint kinase 1 (CHK1) phosphorylation by binding ataxiatelangiectasia mutated and Rad3 related (ATR), a serine/threonine-specific protein kinase, promoted GC apoptosis, and mediated oxaliplatin sensitivity.Conclusion::In general, we discovered a tumor suppressor molecule RP11-789C1.1 and confirmed its mechanism of action, providing a theoretical basis for targeted GC therapy.
6.Research progress of large-channel spinal endoscopic posterior lumbar interbody fusion in the treatment of lumbar degenerative diseases
Lin SUN ; Xiaofeng LI ; Zeyu HUANG ; Yingjie WEI ; Ji MA ; Bo SHI ; Lijun LI
International Journal of Surgery 2024;51(7):499-504
Lumbar interbody fusion is a surgical method for treating lumbar degenerative diseases. By establishing the stability of the lumbar segment, it solves the related symptoms caused by lumbar degenerative diseases. Minimally invasive transforaminal lumbar interbody fusion(MIS-TLIF) is a mature technology for treating lumbar degenerative diseases and improving the stability of the lumbar segment. In recent years, the emergence of lumbar interbody fusion under the small-channel working tube has made it have more minimally invasive characteristics compared to MIS-TLIF, with smaller incisions, less bleeding, and shorter recovery time. However, due to its long operation time, low efficiency of endplate treatment, and high complications incidence rate, it has not been widely popularized. At present, the large-channel endoscopic system, because of its larger field of view under the endoscope and more efficient endoscopic operation tools, reduces the operation time, improves the efficiency of endplate treatment, and reduces the postoperative related complications incidence rate. According to the surgical approach, it can be divided into transforaminal approach, posterior approach, oblique anterior approach, etc. According to the channel mode, it can also be divided into uniaxial endoscopy and unilateral dual-channel endoscopy, and each has its own advantages and disadvantages. Nowadays, the safety and effectiveness of spinal endoscopic posterior lumbar interbody fusion(Endo-PLIF) under the large-channel have achieved satisfactory results. This article reviews the research progress of Endo-PLIF under the large-channel, including surgical indications and contraindications, anatomical basis, surgical techniques, the choice of cages, the choice of fixation methods, safety and effectiveness, advantages and disadvantages, and explores its clinical application and prospects.
7.Comparison on radiation doses of in vitro pre-fenestration and in situ fenestration thoracic endovascular aortic repair in treatment of aortic disease
Fang XUE ; Xiaofeng HAN ; Gang WANG ; Lei SHAO ; Guangrui LIU ; Tiezheng LI ; Xi GUO ; Wei QIU ; Xiaohai MA
Chinese Journal of Interventional Imaging and Therapy 2024;21(9):553-556
Objective To compare the radiation dose of in vitro pre-fenestration and in situ fenestration thoracic endovascular aortic repair(TEVAR)in treatment of aortic disease.Methods Data of 51 patients with aortic diseases who received in vitro pre-fenestration(group A)and 21 cases who underwent in situ fenestration(group B)TEVAR were retrospectively analyzed.The fluoroscopy duration,total reference air kerma(AK),total dose area product(DAP)and TEVAR time were compared between groups.Results TEVAR was successfully completed in all 72 patients.Fluoroscopy duration([21.42±8.04]min vs.[34.57±9.07]min)and total DAP(44315.0[31157.0,56307.5]μGy·m2 vs.72153.0[45460.0,82354.0]μGy·m2)in group A were both significantly lower than those in group B(both P<0.05),while total AK(2423[1638,3533]mGy vs.3600[1898,3921]mGy)and TEVAR time([83.41±22.89]min vs.[81.00±22.13]min)in group A were not significant different from those in group B(both P>0.05).Conclusion Compared with in situ fenestration TEVAR,both the fluoroscopy time and total DAP of in vitro pre-fenestration TEVAR significantly reduced for treating aortic diseases.
8.Assessment of malnutrition diagnosed with GLIM criteria in liver cirrhosis patients and its influencing factors
Shenhui LUO ; Lini WEI ; Qun XIANG ; Xiaofeng SHI ; Juan WANG ; Xuefeng LI
Chinese Journal of Clinical Nutrition 2024;32(4):232-238
Objective:To evaluate the incidence of malnutrition in hospitalized patients with liver cirrhosis using the Global Leadership Initiative on Malnutrition (GLIM) criteria, and to identify the risk factors of malnutrition.Methods:A total of 305 inpatients with liver cirrhosis were included from the Department of Gastroenterology of the People's Hospital of Xiangxi Tujia and Miao Autonomous Prefecture between January 1, 2023 to September 30, 2023. The patient characteristics, clinical indicators, complications, unplanned readmission within 90 days and death outcomes were collected. Malnutrition was diagnosed according to the GLIM criteria, the differences in various indicators were compared between the malnutrition group and the well-nourished group, and the influencing factors of malnutrition were analyzed. The relationship between malnutrition and short-term prognosis was also discussed.Results:Among the enrolled patients, 188 (61.6%) were diagnosed with malnutrition per GLIM criteria, of whom 18 were with Child-Turcotte-Pugh (CTP) Grade A liver function, 108 Grade B, and 62 Grade C. Compared with the well-nourished group, the malnutrition group showed prolonged length of hospital stay ( P<0.001), lower body mass index P=0.003), lower levels of red blood cell count, hemoglobin, serum albumin, serum albumin/globulin ratio, and serum calcium ( P<0.001), higher level of aspartate aminotransferase ( P=0.025), and increased levels of C-reactive protein, total bilirubin, international normalized ratio, plasma prothrombin time and model for end-stage liver disease score ( P<0.001). The proportion of patients with ascites, esophageal gastric varices, upper gastrointestinal bleeding, hepatic encephalopathy and unplanned readmission within 90 days were significantly increased ( P<0.001) in malnutrition group, and the mortality within 90 days was also higher ( P=0.042). Logistic regression analysis identified lower body mass index level ( OR=0.910, P=0.038), ascites ( OR=0.065, P<0.001) and upper gastrointestinal bleeding ( OR=0.184, P=0.001) as the influencing factors of malnutrition in liver cirrhosis patients. Conclusions:The prevalent malnutrition in patients with liver cirrhosis may affect the short-term prognosis. Lower body mass index, ascites and upper gastrointestinal bleeding are influencing factors for malnutrition in liver cirrhosis patients. Therefore, patients with liver cirrhosis should be screened for malnutrition to allow timely nutritional intervention measures.
9.Immune effect of H9N2 subtype AIV NP protein by prokaryotic expression
Xiaofeng LI ; Zhixun XIE ; Zhihua RUAN ; Meng LI ; Dan LI ; Minxiu ZHANG ; Zhiqin XIE ; Sisi LUO ; You WEI ; Liji XIE ; Tingting ZENG ; Yanfang ZHANG ; Jiaoling HUANG ; Sheng WANG
Chinese Journal of Veterinary Science 2024;44(6):1113-1119
The aim of this study is to investigate the immune effect of H9 subtype avian influenza virus(AIV)NP protein on mice and lay the foundation for the development of avian influenza vi-rus(AIV)vaccine.The H9N2 virus NP gene amplification product was cloned into the pET-32a expression vector,and the protein expression was verified by SDS-PAGE and Western blot,and the immune effect was evaluated by measuring the secretion of supernatant multicytokines in mouse splenocytes culture.The results showed that the total length of the coding region sequence of NP gene was 1 497 bp,NP recombinant proteins exist in both soluble and insoluble protein forms,and the specific bands were visible in Western blot.After immunizing mice,serum produces IgG-bind-ing antibodies with antibody titers of 1∶40 000.Compared with the control group,IL-2,IL-5 and IL-13 were significantly increased(P<0.001),and the secretion of IL-6 was significantly increased compared with the control group.IL-4 and IL-12 p70 secretions were elevated compared with con-trols,but there was no significant difference.Compared with the control group,the secretions of IL-1β,IL-18,GM-CMF,TNF-α and IFN-γ were inhibited,but the difference was not significant(P>0.05).The results showed that NP recombinant protein is a good immunogen,laying a foundation for in-depth research on influenza vaccine.
10.Effect of permissible hypercapnia combined with remote ischemic preconditioning on brain oxygen saturation and postoperative cognition in patients undergoing thoracoscopic lung cancer surgery
Wei LU ; Danyan ZHU ; Xiaofeng JIANG ; Xiao RAO ; Wenyong PENG
China Modern Doctor 2024;62(14):19-23
Objective To explore the effects of remote ischemic preconditioning combined with permissive hypercapnia on brain oxygen saturation and postoperative cognition in patients which undergoing thoracoscopic lung cancer surgery.Methods A collection of 64 patients elective requiring thoracoscopic lung cancer surgery who were divided into control group and combined group according to the randomized grouping method,with 32 cases in each group.The PaCO2 in the control group of patient was maintained at normal,and patients in the combination group were given permissive hypercapnia ventilation strategies and performed remote ischemic preconditioning,PaCO2 is maintained at 45-50mmHg(1mmHg=0.133kPa).Record the cerebral oxygen saturation(rSO2)at the five time points before operation(T0),10min after one lung ventilation(T1),30min after one lung ventilation(T2),10 min after lung recruitment(T3)and the end of surgery(T4),measured the internal jugular venous blood oxygen saturation(SjvO2)and calculated cerebral arteriovenous blood oxygen content difference(CaO2-CjvO2),brain oxygen uptake rate(CERO2).Monitored the average arterial pressure(MAP)and heart rate(HR)of the hemodynamic indicators at the above five time points.The scores of cognitive function were recorded 1 day before operation and 1 day and 3 days after operation;detected the levels of serum neuron-specific enolase(NSE),amyloid β(Aβ)and S100β protein(S100β)in 1 day before surgery,24hours after surgery and 48hours after surgery;Comparison of postoperative related indicators and adverse reactions between the patients of two groups.Results The rSO2 and SjvO2 of combined group were higher than control group in the T1-T4,but CaO2-CjvO2 and CERO2 were lower than those of control group.There was no significant difference in HR and MAP between two groups from T0-T4.The mini-mental state examination(MMSE)score of the combined group was significantly higher than that of the control group on the 1 day after operation.The level of serum NSE,Aβ and S100β in the combined group was lower than those of control group at 24hours and 48hours after operation.There was no significant difference in incidence of postoperative adverse reactions and postoperative related indexes between the two groups.Conclusion Permissive hypercapnia combined with remote ischemic preconditioning can increase cerebral oxygen saturation in patients undergoing thoracoscopic lung cancer surgery,improve cerebral oxygen metabolism and reduce the levels of serum neuron-specific enolase,β-amyloid protein and S100β protein,decrease the postoperative cognitive dysfunction.

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