OBJECTIVE To explore the effectiveness and possible mechanism of Shangke Lengtong Patch in treating knee osteo-arthritis with cold-dampness arthralgia obstruction.METHODS A total of 70 patients who met the inclusion criteria of knee osteoar-thritis with cold-dampness arthralgia obstruction in the Orthopedics Department of Affiliated Hospital of Nanjing University of Chinese Medicine from November to December 2024 were randomly divided into an experimental group and a control group,with 35 cases in each group.During the treatment,1 case dropped out of the experimental group,3 cases dropped out of the control group,and 1 case was discontinued.The experimental group was treated with Shangke Lengtong Patch,and the control group was treated with Compound Nanxing Zhitong Ointment.The WOMAC scores and TCM syndrome scores of the two groups before and after treatment were compared to comprehensively evaluate the clinical efficacy.The changes in the expression levels of CGRP,substance P,HMGB1,IL-1β,CX-CL12,and CXCR4 in the serum of the two groups were detected by ELISA.RESULTS After 3,7,and 14 d of treatment,the WOMAC scores and TCM syndrome scores of the two groups were significantly reduced(P<0.05,P<0.01),and the score of aggrava-ted cold in the experimental group was better than that in the control group at 7 d of treatment(P<0.05);after 14 d of treatment,the expression levels of CGRP,substance P,HMGB1,IL-1β,CXCL12,and CXCR4 in the serum of the two groups were significantly re-duced(P<0.05),and there was no statistical difference between the two groups.CONCLUSION Shangke Lengtong Patch can sig-nificantly relieve the pain symptoms of knee osteoarthritis patients with cold-dampness arthralgia obstruction and improve the joint func-tion of patients.It may improve synovial inflammation by inhibiting the HMGB1/CXCL12/CXCR4 pathway,thereby exerting a thera-peutic effect.

OBJECTIVE To explore the intervention mechanism of Shangke Lengtongtie on cold hyperalgesia in KOA mice based on the HMGB1/CXCL12/CXCR4 signaling axis.METHODS Monosodium iodoacetate(MIA)was used for the intra-articular injec-tion into the knee joint to establish mice model of knee osteoarthritis(KOA).Peripheral blood monocytes were extracted from mice,cultured,and then reinfused into the tail vein of the mice.Subsequently,in vivo animal imaging was used to observe the recruitment sites of these monocytes.The cold hyperalgesia threshold was measured at various time points in each group of mice.Hematoxylin and eosin(HE)staining was used to evaluate the level of synovial pathological changes.ELISA was employed to detect the expression of in-flammatory factors IL-1β,TNF-α,and pain mediators CGRP and Substance P in mouse serum.Western blot and qPCR methods were used to detect the protein and gene expression of cold hyperalgesia-related indicators such as TRPA1,TRPM8,HMGB1,CXCL12,CXCR4,Collagen Ⅰ,and Netrin-1 in synovial tissue,as well as DCC in dorsal root ganglia(DRG)tissue.RESULTS In vivo ima-ging showed that after the monocytes were reinfused into KOA mice,they were recruited to the knee joint area,with the HMGB1 group exhibiting a greater recruitment of circulating monocytes at the knee joint.Additionally,compared to the control group,the KOA group and HMGB1 group showed inflammatory pathological changes in the synovium,increased expression of serum inflammatory factors and pain mediators,reduced cold hyperalgesia threshold,and upregulated protein and gene expression of cold hyperalgesia-related indica-tors in synovial and DRG tissues.The changes were more significant in the HMGB1 group compared to the KOA group(P<0.05).Af-ter treatment with Shangke Lengtongtie or GL intervention,synovial inflammation was alleviated,serum inflammatory factors and pain mediators decreased,cold hyperalgesia threshold increased,and the upregulation of cold hyperalgesia-related indicator protein and gene expression levels was significantly reversed(P<0.05).CONCLUSION Shangke Lengtongtie exerts a beneficial effect on the mitigation of synovitis and cold hyperalgesia in KOA mice,a therapeutic mechanism that possibly mediated through the inhibition of the HMGB1/CXCL12/CXCR4 signaling axis.

OBJECTIVE To explore the effectiveness and possible mechanism of Shangke Lengtong Patch in treating knee osteo-arthritis with cold-dampness arthralgia obstruction.METHODS A total of 70 patients who met the inclusion criteria of knee osteoar-thritis with cold-dampness arthralgia obstruction in the Orthopedics Department of Affiliated Hospital of Nanjing University of Chinese Medicine from November to December 2024 were randomly divided into an experimental group and a control group,with 35 cases in each group.During the treatment,1 case dropped out of the experimental group,3 cases dropped out of the control group,and 1 case was discontinued.The experimental group was treated with Shangke Lengtong Patch,and the control group was treated with Compound Nanxing Zhitong Ointment.The WOMAC scores and TCM syndrome scores of the two groups before and after treatment were compared to comprehensively evaluate the clinical efficacy.The changes in the expression levels of CGRP,substance P,HMGB1,IL-1β,CX-CL12,and CXCR4 in the serum of the two groups were detected by ELISA.RESULTS After 3,7,and 14 d of treatment,the WOMAC scores and TCM syndrome scores of the two groups were significantly reduced(P<0.05,P<0.01),and the score of aggrava-ted cold in the experimental group was better than that in the control group at 7 d of treatment(P<0.05);after 14 d of treatment,the expression levels of CGRP,substance P,HMGB1,IL-1β,CXCL12,and CXCR4 in the serum of the two groups were significantly re-duced(P<0.05),and there was no statistical difference between the two groups.CONCLUSION Shangke Lengtong Patch can sig-nificantly relieve the pain symptoms of knee osteoarthritis patients with cold-dampness arthralgia obstruction and improve the joint func-tion of patients.It may improve synovial inflammation by inhibiting the HMGB1/CXCL12/CXCR4 pathway,thereby exerting a thera-peutic effect.

Objective:To explore the correlation between serum carbohydrate antigen 72-4 (CA72-4) level and pathological differentiation degree of gastric cancer, and to provide reference for early diagnosis, prognosis evaluation and efficacy monitoring of gastric cancer.Methods:A retrospective study was conducted on 136 patients with gastric cancer admitted to Zhejiang Provincial People′s Hospital from April 2022 to December 2024. According to postoperative pathological diagnosis, patients were divided into well-differentiated group ( n=9), moderately differentiated group ( n=36) and poorly/undifferentiated group ( n=91). Preoperative CA72-4 levels and clinical characteristics of patients were collected to analyze the relationship between differentiation degree and CA72-4 level, and to compare the changes of CA72-4 before and after surgery in gastric cancer patients with different differentiation degrees. Results:The preoperative CA72-4 level in the poorly/undifferentiated group was significantly higher than that in the well-differentiated group and moderately differentiated group (all P<0.001). Preoperative CA72-4 level in gastric cancer patients was weakly negatively correlated with differentiation degree ( R=-0.341). After surgery, the CA72-4 level in the poorly/undifferentiated group decreased significantly compared with that before surgery ( P=0.043), while the changes in the well-differentiated group and moderately differentiated group were not significant. Conclusions:There are significant differences in CA72-4 levels among gastric cancer groups with different differentiation degrees. Preoperative CA72-4, as a tumor marker, is of great significance for early diagnosis, prognosis evaluation and postoperative monitoring of gastric cancer. Especially in patients with poorly/undifferentiated gastric cancer, the CA72-4 level is relatively high, which can be used for auxiliary diagnosis and formulation of treatment plans.

OBJECTIVE To explore the intervention mechanism of Shangke Lengtongtie on cold hyperalgesia in KOA mice based on the HMGB1/CXCL12/CXCR4 signaling axis.METHODS Monosodium iodoacetate(MIA)was used for the intra-articular injec-tion into the knee joint to establish mice model of knee osteoarthritis(KOA).Peripheral blood monocytes were extracted from mice,cultured,and then reinfused into the tail vein of the mice.Subsequently,in vivo animal imaging was used to observe the recruitment sites of these monocytes.The cold hyperalgesia threshold was measured at various time points in each group of mice.Hematoxylin and eosin(HE)staining was used to evaluate the level of synovial pathological changes.ELISA was employed to detect the expression of in-flammatory factors IL-1β,TNF-α,and pain mediators CGRP and Substance P in mouse serum.Western blot and qPCR methods were used to detect the protein and gene expression of cold hyperalgesia-related indicators such as TRPA1,TRPM8,HMGB1,CXCL12,CXCR4,Collagen Ⅰ,and Netrin-1 in synovial tissue,as well as DCC in dorsal root ganglia(DRG)tissue.RESULTS In vivo ima-ging showed that after the monocytes were reinfused into KOA mice,they were recruited to the knee joint area,with the HMGB1 group exhibiting a greater recruitment of circulating monocytes at the knee joint.Additionally,compared to the control group,the KOA group and HMGB1 group showed inflammatory pathological changes in the synovium,increased expression of serum inflammatory factors and pain mediators,reduced cold hyperalgesia threshold,and upregulated protein and gene expression of cold hyperalgesia-related indica-tors in synovial and DRG tissues.The changes were more significant in the HMGB1 group compared to the KOA group(P<0.05).Af-ter treatment with Shangke Lengtongtie or GL intervention,synovial inflammation was alleviated,serum inflammatory factors and pain mediators decreased,cold hyperalgesia threshold increased,and the upregulation of cold hyperalgesia-related indicator protein and gene expression levels was significantly reversed(P<0.05).CONCLUSION Shangke Lengtongtie exerts a beneficial effect on the mitigation of synovitis and cold hyperalgesia in KOA mice,a therapeutic mechanism that possibly mediated through the inhibition of the HMGB1/CXCL12/CXCR4 signaling axis.

Objective:To explore the correlation between serum carbohydrate antigen 72-4 (CA72-4) level and pathological differentiation degree of gastric cancer, and to provide reference for early diagnosis, prognosis evaluation and efficacy monitoring of gastric cancer.Methods:A retrospective study was conducted on 136 patients with gastric cancer admitted to Zhejiang Provincial People′s Hospital from April 2022 to December 2024. According to postoperative pathological diagnosis, patients were divided into well-differentiated group ( n=9), moderately differentiated group ( n=36) and poorly/undifferentiated group ( n=91). Preoperative CA72-4 levels and clinical characteristics of patients were collected to analyze the relationship between differentiation degree and CA72-4 level, and to compare the changes of CA72-4 before and after surgery in gastric cancer patients with different differentiation degrees. Results:The preoperative CA72-4 level in the poorly/undifferentiated group was significantly higher than that in the well-differentiated group and moderately differentiated group (all P<0.001). Preoperative CA72-4 level in gastric cancer patients was weakly negatively correlated with differentiation degree ( R=-0.341). After surgery, the CA72-4 level in the poorly/undifferentiated group decreased significantly compared with that before surgery ( P=0.043), while the changes in the well-differentiated group and moderately differentiated group were not significant. Conclusions:There are significant differences in CA72-4 levels among gastric cancer groups with different differentiation degrees. Preoperative CA72-4, as a tumor marker, is of great significance for early diagnosis, prognosis evaluation and postoperative monitoring of gastric cancer. Especially in patients with poorly/undifferentiated gastric cancer, the CA72-4 level is relatively high, which can be used for auxiliary diagnosis and formulation of treatment plans.

OBJECTIVES: To investigate the effect of nuclear protein (Nupr) 1 on the pathological progression of osteoarthritis and its relationship with ferroptosis of chondrocytes. METHODS: Chondrocytes from mouse knees were divided into small interfering RNA (siRNA) control group, small interfering RNA targeting Nupr1 (siNupr1) group, siRNA control+IL-1β group (siRNA control interference for 24 h followed by 10 ng/mL IL-1β) and siNupr1+IL-1β group (siNupr1 interference for 24 h followed by 10 ng/mL IL-1β). The protein and mRNA expressions of Nupr1 were detected by Western blotting and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cell proliferation viabilities were measured using the cell counting kit-8 method. The levels of ferrous ions were detected by FerroOrange staining. Lipid peroxidation levels were detected by C11-BODIPY-591 fluorescence imaging. The contents of malondialdehyde (MDA) and glutathione (GSH) were detected by enzyme-linked immunosorbent assay. The protein expressions of acyl-CoA synthetase long-chain family (ACSL) 4, P53, glutathione peroxidase (GPX) 4 and solute carrier family 7 member 11 gene (SLC7A11) were detected by Western blotting. The osteoarthritis model was constructed by destabilization of the medial meniscus (DMM) surgery in 7-week-old male C57BL/6J mice. The mice were randomly divided into four groups with 10 animals in each group: sham surgery (Sham)+adeno-associated virus serotype 5 (AAV5)-short hairpin RNA (shRNA) control group, Sham+AAV5-shRNA control targeting Nupr1 (shNupr1) group, DMM+AAV5-shRNA control group, and DMM+AAV5-shNupr1 group. Hematoxylin and eosin staining and Safranin O-Fast Green staining were used to observe the morphological changes in cartilage tissue. The Osteoarthritis Research Society International (OARSI) osteoarthritis cartilage histopathology assessment system was used to evaluate the degree of cartilage degeneration in mice. The mRNA expressions of matrix metallopeptidase (MMP) 13, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) 5, cyclooxy-genase (COX) 2, and GPX4 were detected by qRT-PCR. RESULTS: In vitro experiments showed that knocking down Nupr1 alleviated the decrease of chondrocyte proliferation activity induced by IL-1β, reduced iron accumulation in mouse chondrocytes, lowered lipid peroxidation, downregulated ACSL4 and P53 protein expression and upregulated GPX4 and SLC7A11 protein expression (all P<0.01), thereby inhibiting ferroptosis in mouse chondrocytes. Meanwhile, in vivo animal experiments demonstrated that knocking down Nupr1 delayed the degeneration of articular cartilage in osteoarthritis mice, improved the OARSI score, slowed down the degradation of the extracellular matrix in osteoarthritis cartilage, and reduced the expression of the key ferroptosis regulator GPX4 (all P<0.01). CONCLUSIONS Knockdown of Nupr1 can delay the pathological progression of osteoarthritis through inhibiting ferroptosis in mouse chondrocytes.
Animals ; Ferroptosis ; Mice ; Chondrocytes/metabolism* ; Osteoarthritis/pathology* ; RNA, Small Interfering/genetics* ; Basic Helix-Loop-Helix Transcription Factors/genetics* ; Interleukin-1beta/metabolism* ; Phospholipid Hydroperoxide Glutathione Peroxidase/genetics* ; Coenzyme A Ligases/genetics* ; Tumor Suppressor Protein p53/metabolism* ; Mice, Inbred C57BL ; DNA-Binding Proteins ; Neoplasm Proteins ; Amino Acid Transport System y+ ; Nuclear Receptor Subfamily 1, Group D, Member 1

Objective To compare short-term therapeutic outcomes and the safety of percutane-ous radiofrequency ablation (PRFA) with left single lung ventilation (LSLV) for liver cancer of the hepatic dome (LCHD) and that of PRFA for right liver carcinoma in favorable location. Methods Thirty one patients with hepatocellular carcinoma (belonging to LCHD) receiving PRFA with LSLV (Group LCHD) between January 2006 and January 2009 in our hospital were selected, and 45 control patients with right lobe HCC ≥1 cm away from the liver capsule, gallbladder, and main portal bran-ches were also included. One month after PRFA, residual tumors were followed up with contrast en-hanced CT and alpha fetal protein and PRFA was repeated in the presence of residual foci. Tumor-free survival time was defined as the duration from complete ablation to diagnosed local tumor progression.The Mann-Whitney test was used to compare age, tumor diameter, and average number of punctures between LCHD patients and controls. A χ2 test was used for comparison of the incidence of complica-tions and incomplete tumor ablation rate. The Kaplan-Meier's method was used for calculation of local tumor-free survival rate compared with a log-rank test. Results The incidence of right shoulder pain was significantly higher in LCHD patients than in controls (87. 1% vs 11. 1%, P<0. 01). LCHD pa-tients showed no difference from controls in the average number of punctures (2. 8±. 5 vs 3. 2±. 5,P>0. 05). Meanwhile, there was no difference between the 2 groups in average duration of treatment and hospitalization, and the complete tumor ablation rate at first PRFA. No differences were observed in the 1-, 2- and 3-year local tumor-free survival rates between LCHD patients (85. 5% , 65. 8% , and 36. 4% ,respectively) and controls (87.7%, 62. 3% , and 34.0% , respectively). Conclusion PRFA with LSLV for LCHD seems to promise comparable short-term outcomes and safety to PRFA for right liver carcinoma of fa-vorable location and should be preferred as one of the therapeutic options for LCHD patients with tumor di-ameters≤5 cm regardless of its unique location.

Objective To analyze and evaluate the effect addition of albumin to the extracorporeal circulation (CPB) in patients undergoing valve replacement. Methods 62 patients under 60 years of age, with the blood level of albumin nearly normal, undergoing mitral valve replacement or aortic valve replacement were randomly divided into two groups. In 34 patients albumin was added to the priming fluid of extraporeal circulation, and in 28 patients it was not. The pre-operative and postoperative serum albumin levels, the duration of assisting ventilation, and the amount of albumin needed between the time of operation to 7 am of the first postoperative day were compared. In both groups the primary priming fluid consisted of balanced electrolyte solution, hydroxyethyl starch, 5% sodium bicarbonate, and 25% mannitol. Results All the indexes, including the preoperative level of albumin, the amount of albumin needed from operation to 7 am of the first postoperative day, and the albumin level at 7 am of the first postoperative day showed no notable differences. Conclusion For patients with no hypoalbuminemia, mitral valve replacement or aortic valve replacement is safe to withhold the addition of albumin to the priming fluid for CPB.