ObjectiveTo investigate the possible mechanism of Jishe Qushi Capsule （脊蛇祛湿胶囊， JQC） in treating rheumatoid arthritis （RA） from the perspective of macrophage polarization mediated by neutrophil extracellular traps （NETs）. MethodsTwenty-four female SD rats were randomly divided into four groups， blank control group， model group， JQC group， and peptidylarginine deiminase 4 （PAD4） inhibitor group with 6 rats in each group. All groups but the blank control group were subjected to the induction of collagen-induced arthritis （CIA）. After successful model establishment， rats in the JQC group received intragastric administration of JQC 1.47 g/kg daily； rats in the PAD4 inhibitor group received intraperitoneal injections of the PAD4 inhibitor 4 mg/kg weekly. Rats in the blank， model， and PAD4 inhibitor groups received 2 ml of pure water daily by gavage. All treatments lasted 4 weeks. Joint lesions of each group were assessed on day 7， 14， 21， 28， and 35 after model establishment， and arthritis index （AI） scores were recorded. At 24 h after the final administration， histopathology of knee joints， including HE staining， safranin O-fast green staining， and TRAP staining， was performed. Flow cytometry was used to detect the counts of M1 and M2 macrophages in peripheral blood. ELISA was used to determine serum levels of TRACP， NETs， TNF-α， IL-1β， and iNOS. Western Blotting and qRT-PCR were used to measure MPO， NE， RANKL， OPG， and p65 protein and mRNA expression in knee cartilage tissue. ResultsCompared with the blank control group， the model group showed increased AI scores （P＜0.05）， marked synovial inflammatory infiltration， angiogenesis， and bone-cartilage destruction， increased TRAP-positive osteoclasts， increased M1 macrophages and decreased M2 macrophages， elevated serum TRACP， NETs， TNF-α， IL-1β， and iNOS （P＜0.05）， elevated MPO， NE， RANKL， and p65 protein/mRNA expression and decreased OPG protein/mRNA expression in knee cartilage tissue （P＜0.05）. Compared with the model group， the JQC group exhibited improved synovial inflammation， angiogenesis， and bone-cartilage damage， reduced AI scores on day 21， 28， and 35， decreased osteoclast counts， decreased M1 macrophages and increased M2 macrophages， reduced serum TRACP， NETs， TNF-α， IL-1β， and iNOS （P＜0.05）， decreased MPO， NE， RANKL， and p65 protein/mRNA expression and increased OPG expression （P＜0.05）. Compared with the PAD4 inhibitor group， the JQC group showed significantly lower AI scores， reduced M1 macrophages， increased M2 macrophages （P＜0.05）， reduced serum TRACP， TNF-α， IL-1β， and iNOS， decreased MPO， RANKL， and p65 expression， and increased OPG levels （P＜0.05）. ConclusionThe therapeutic mechanism of JQC for RA may involve inhibition of NETs formation， downregulation of the RANKL/NF-κB signaling pathway， and regulation of macrophage M1/M2 polarization imbalance， thereby suppressing osteoclastogenesis and inflammatory bone destruction.

OBJECTIVES: To analyze the molecular mechanism of Xixian Pills for treatment of rheumatoid arthritis (RA). METHODS: Forty-eight rats were randomized into 6 groups (n=8), including a normal control group, a collagen-induced arthritis (CIA) model group, 3 Xixian Pills treatment (200, 400 and 800 mg/kg) groups, and a Tripterygium glycosides tablet (TGT) treatment group. In the latter 4 groups, the rats were treated with daily gavage of Xixian Pills or TGT 2 weeks after CIA modeling for 3 consecutive weeks. The differentially expressed proteins in high-dose Xixian Pills group and the model group compared with the normal control group were screened based on the tandem mass spectrometry tag (TMT) technology, and the core targets and signaling pathways were analyzed. The immune cell infiltration and gene expression data were analyzed using ggplot2 and tidyverse packages, and the correlation coefficients between the core targets and the immune cells were calculated. RESULTS: The CIA rats showed significantly increased serum levels of TNF-α and IL-6 and lowered serum IL-10 level. Treatments with high- and medium-dose Xixian Pills and TGT all significantly reduced serum TNF‑α and IL-6 and increased IL-10 levels in CIA rats. Proteomic analysis identified 160 differential proteins between the model group and high-dose Xixian Pills group, and the core targets included CCL5, STAT1, GZMB and IL7R. The areas under the ROC curve of CCL5 and STAT1 were both greater than 0.9. Immunohistochemical and immunofluorescence staining revealed increased levels of CCL5 and STAT1 in the ankle joints of CIA rats, which were significantly decreased after treatment with Xixian Pills. CONCLUSIONS Treatment with Xixian Pills offers protection of the joints in CIA rats possibly by inhibiting joint inflammation via regulating protein expressions of CCL5 and STAT1.
Animals ; Drugs, Chinese Herbal/pharmacology* ; Rats ; Arthritis, Rheumatoid/metabolism* ; Proteomics ; Tripterygium/chemistry* ; Arthritis, Experimental/metabolism* ; Tumor Necrosis Factor-alpha/blood* ; Interleukin-10/blood* ; Interleukin-6/blood* ; Male ; Rats, Sprague-Dawley ; Signal Transduction

OBJECTIVE: To investigate the clinical application of high-frequency color Doppler ultrasound (HFCDU) in detecting perforators in the deep adipose layers for harvesting super-thin anterolateral thigh flap (ALTF). METHODS: Between August 2019 and January 2023, 45 patients (46 sides) with skin and soft tissue defects in the foot and ankle were treated, including 29 males and 16 females, aged from 22 to 62 years, with an average of 46.7 years. The body mass index ranged from 19.6 to 36.2 kg/m ², with an average of 23.62 kg/m ². The causes of injury included traffic accident injury in 15 cases, heavy object crush injury in 20 cases, mechanical injury in 8 cases, heat crush injury in 1 case, and chronic infection in 1 case. There were 20 cases on the left side, 24 cases on the right side, and 1 case on both sides. After thorough debridement, the wound size ranged from 5 cm×4 cm to 17 cm×11 cm. All patients underwent free super-thin ALTF transplantation repair. HFCDU was used to detect the location of the perforators piercing the deep and superficial fascia, as well as the direction and branches of the perforators within the deep adipose layers before operation. According to the preoperative HFCDU findings, the dimensions of the super-thin ALTF ranged from 6 cm×4 cm to 18 cm×12 cm. The donor sites of the flaps were directly sutured. RESULTS: A total of 55 perforators were detected by HFCDU before operation, but 1 was not found during operation. During operation, a total of 56 perforators were found, and 2 perforators were not detected by HFCDU. The positive predictive value of HFCDU for identifying perforator vessels was 98.2%, and the sensitivity was 96.4%. Among the 54 perforators accurately located by HFCDU, the orientation of the perforators in the deep adipose layers was confirmed during operation. There were 21 perforators (38.9%) traveled laterally and inferiorly, 12 (22.2%) traveled medially and inferiorly, 14 (25.9%) traveled laterally and superiorly, 5 (9.3%) traveled medially and superiorly, and 2 (3.7%) ran almost vertically to the body surface. Among the 54 perforators accurately located by HFCDU, 35 were identified as type 1 perforators and 12 as type 2 perforators (HFCDU misidentified 7 type 2 perforators as type 1 perforators). The sensitivity of HFCDU in identifying type 1 perforators was 100%, with a positive predictive value of 83.3%. For type 2 perforators, the sensitivity was 63.2%, and the positive predictive value was 100%. The surgeries were successfully completed. The super-thin ALTF had a thickness ranging from 2 to 6 mm, with an average of 3.56 mm. All super-thin ALTF survived, however, 1 flap experienced a venous crisis at 1 day after operation, but it survived after emergency exploration and re-anastomosis of the veins; 1 flap developed venous crisis at 3 days after operation but survived after bleeding with several small incisions; 3 flaps had necrosis at the distal edge of the epidermis, which healed after undergoing dressing changes. All 45 patients were followed up 6-18 months (mean, 13.6 months). Three flaps required secondary defatting procedures, while the rest had the appropriate thickness, and the overall appearance was satisfactory. CONCLUSION Preoperative application of HFCDU to detect the perforator in the deep adipose layers can improve the success and safety of the procedure by facilitating the harvest of super-thin ALTF.
Male ; Female ; Humans ; Thigh/surgery* ; Plastic Surgery Procedures ; Prospective Studies ; Skin Transplantation ; Free Tissue Flaps ; Burns ; Soft Tissue Injuries/surgery* ; Ultrasonography, Doppler, Color ; Crush Injuries/surgery* ; Perforator Flap ; Treatment Outcome

Objective: To investigate the changes of microorganisms and metabolites in joint effusion of patients with Rheumatoid arthritis(RA), Osteoarthritis(OA) and Urarthritis(UA). To provide new ideas for the study of the effect of microbiota on the pathogenesis of arthritis. Methods: Joint effusion samples were collected from 20 patients with RA, 20 patients with OA, and 20 patients with UA. 16S rRNA gene sequencing and untargeted ultra-high performance Liquid chromatography-mass spectrometry(LC-MS) were used to explore the differences in microorganisms and metabolites among the three groups. Pearson correlation analysis was used to detect the correlation between effusion microbiota and metabolites. Results: There were differences in microbial diversity and microbiota composition among the three groups. Combined with VIP>1 from OPLS-DA andP<0.05 from two-tailed Students t-test, 45 differential metabolites(Between RA and OA groups), 38 differential metabolites(Between UA and OA groups) and 16 differential metabolites(Between RA and UA groups), were identified. GO analysis and KEGG pathway analysis showed that the differential metabolic pathways among the three groups were mainly concentrated in citric acid cycle(TCA cycle), nucleotide metabolism, amino acid metabolism and glycolysis pathway. Correlation analysis of joint effusion microbiota and metabolites suggested that bacteria enriched in the three groups of joint effusion, such asPrevotella,Clostridium ruminosus,Prevotellaceae＿UCG-001, were related to many key metabolites such as lysozyme, uric acid, glucose, and L-glutamine. Conclusion This study shows that there are a variety of bacterial flora in joint cavity effusion of RA, OA, and UA patients, and the differential metabolites produced by them are involved in the pathogenesis of the three types of arthritis by affecting a variety of metabolic pathways.

Objective To use the GEO dataset and bioinformatics techniques,such as LASSO logistic regression,ssGSEA,and WGCNA,to screen for RA diagnostic markers and investigate the impact of earthly flavonoids in Smi-lax glabra Roxb.on specific immune cell infiltration,to screen for rheumatoid arthritis(RA)diagnostic markers on specific immune cell infiltration and to analyze the combination of flavonoids in Smilax glabra Roxb.and diagnostic markers.Methods The normal control group and RA gene chip were obtained from the Gene Expression Omnibus database.The R 4.3.0 WGCNA software package was used to integrate and analyze the dataset,identify co-expres-sion modules and associated trait information,and screen key modules closely related to RA.LASSO regression a-nalysis was performed using the glmnet package in R to identify characteristic genes for RA.The area under the re-ceiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of the characteristic genes in RA.The gene expression data of the normal control group and RA group were subjected to quantitative immune cell infiltration analysis using the GSVA,limma,and GSEABase packages in R.The chemical components of earth-worm flavonoids in Smilax glabra Roxb.were analyzed based on UHPLC-Q-Exactive Orbitrap MS.The correlation between flavonoids and characteristic genes was assessed through molecular docking.Results The LASSO regres-sion algorithm selected 5 characteristic genes(apolipoprotein D,zinc finger and BTB domain containing 16,C-C chemokine receptor type 5,matrix metalloproteinase 1,coronin-1A).The area under ROC curve of all 5 character-istic genes was greater than 0.85,which exhibited positive correlations with various immune cells.Twenty earth-worm flavonoids of Smilax glabra Roxb.were identified using UHPLC-Q-Exactive/MS,and Mulberrin and Neobavaisoflavone were well combined with 5 immune characteristic genes.Conclusion Flavonoids compounds of Smilax glabra Roxb.have good combination with RA immune characteristic genes,providing a scientific basis for RA immunomodulation therapy and early diagnosis.

Objective To observe the value of multimodal models established combined 18F-FDG PET/CT radiomics with clinical data for evaluating response of locally advanced head and neck squamous cell carcinoma(LA-HNSCC)to neoadjuvant immuno-chemotherapy.Methods Totally 213 LA-HNSCC patients were retrospectively enrolled and randomized into training set(n=170)and test set(n=43)at the ratio of 8∶2.Radiomics features of tumors on 18F-FDG PET/CT were extracted and selected from training set,and the independent clinical predictors were screened with sequential univariate and multivariate logistic regressions.Radiomics models,clinical models and combined multimodal models were constructed using different algorithms,including adaptive boosting(AdaBoost),decision tree,naive Bayes,random forest(RF),support vector machine(SVM)and extreme gradient boosting(XGBoost),respectively.The receiver operating characteristic(ROC)curves were drawn,and the area under the curves(AUC)were calculated to assess the efficacy of each model for predicting the response of LA-HNSCC to neoadjuvant immuno-chemotherapy,and the decision curve analysis(DCA)was performed to explore the net benefit of each model.Results Totally 110 radiomics features were selected,and CD4/CD8 ratio was the independent clinical predictor of the response of LA-HNSCC to neoadjuvant immuno-chemotherapy.Models based on AdaBoost and XGBoost algorithms had high and stable efficacy for predicting tumor response to neoadjuvant immuno-chemotherapy,among which the multimodal models had better performance(AUC=0.943,0.930)than radiomics models(AUC=0.939,0.925)and clinical models(AUC=0.903,0.910)in test set(all P＜0.05).Conclusion Multimodal models established combined 18F-FDG PET/CT radiomics with CD4/CD8 ratio were more effective for predicting response of LA-HNSCC to neoadjuvant immuno-chemotherapy than any single model.

Objective:To investigate the dosimetric factors associated with acute nausea and vomiting (RINV) during intensity modulated radiotherapy for nasopharyngeal carcinoma.Methods:General clinical data and organs at risk (OAR) doses from 130 newly diagnosed early nasopharyngeal carcinoma patients who received radiation therapy alone in Henan Cancer Hospital from February 2018 to February 2023 were retrospectively analyzed. Dosimetric parameters were recorded, and the correlation between the parameters and the degree of nausea and vomiting was statistically analyzed using univariate and multivariate logistic regression models.Results:All 130 patients had symptoms of ≥ grade 1 nausea and vomiting. In the comparison of dosimetric parameters between patients with < grade 2 and ≥ grade 2 nausea, except the brainstem V 20 Gy, all parameters showed statistically significant differences (all P<0.05). The inner ear D max, and D max, D mean, V 10 Gy, V 20 Gy, V 30 Gy of the throat, oral cavity, pharyngeal constrictor, dorsal vagal complex (DVC) showed statistically significant differences between patients with grade 1 and grade 2 nausea (all P<0.05). The results of multivariate regression analysis showed that DVC V 30 Gy was a significant influencing factor in predicting the severity of nausea ( OR=73.95, 95% CI: 4.66-1172.60, P<0.001), and there was a significant correlation between oral V 30 Gy and the severity of vomiting ( OR=37.69, 95% CI: 1.26-1125.42, P=0.04). Conclusions:Even if OAR are exposed to lower doses of radiation, nausea or vomiting symptoms can still occur. The occurrence of RINV is significantly associated with DVC and oral radiation doses.

Objective:To investigate the prognosis and influencing factors of different treatment strategies in T 3-T 4 nasal sinus adenocarcinoma. Methods:The data of 93 cases of T 3-T 4 stage nasal sinus adenocarcinoma diagnosed from 2006 to 2018 were retrospectively analyzed. All patients were divided into combined operation group and non-operation group. The survival status and failure mode after corresponding treatment were analyzed. The enumeration data were analyzed by Chi-square test or Fisher's exact test. Survival analysis was performed by Kaplan-Meier method. Univariate analysis was conducted by log-rank test. Multivariate prognostic analysis was performed by Cox model. Results:The average follow-up time in the whole cohort was 81.3 months (18-156 months). By the end of follow-up, a total of 38.7% (36/93) of patients had local recurrence, 14.0% (13/93) had distant metastasis, 17.2% (16/93) had local recurrence complicated with distant metastasis, and 28.0% (26/93) were stable. The overall 2-, 5-, and 10-year overall survival (OS) and progression free survival (PFS) rates were 83.5%, 59.3%, 31.8% and 73.6%, 40.7% and 25.3%, respectively. In univariate analysis, the PFS and OS of patients aged 46-64 years old (all P<0.001), male ( P=0.022, P=0.001), patients with lesions located in the maxillary sinus ( P=0.001, P<0.001), adenoid cystic carcinoma ( P=0.001, P<0.001), non-invasion of orbital / clivus ( P=0.041, P<0.001), GTV P dose>64 Gy ( P=0.003, P=0.006) and N 1 stage ( P=0.014, P=0.014) were statistically different among different treatment modes. Multivariate analysis showed that age ≥65 years old ( P=0.012, P=0.005), orbital / clival invasion ( P<0.001, P=0.005), and GTV p dose ≤64 Gy ( P<0.001, P=0.011) were the independent adverse prognostic factors affecting PFS and OS in T 3-T 4 stage nasal sinus adenocarcinoma. Conclusions:The local failure rate of T 3-T 4 stage nasal sinus adenocarcinoma is high after treatment. Age, orbital / clival invasion, and GTV p dosage are the independent adverse prognostic factors. Surgery based intervention is superior to other treatment strategies.

Objective:To investigate the clinical value of changes in peripheral NKT cells and tumor abnormal proteins (TAPs) in stage Ⅲ-Ⅳ B head and neck squamous cell carcinoma (HNSCC) before and after radiotherapy. Methods:A retrospective analysis was performed using the data of 101 HNSCC patients, who were confirmed from January 2019 to December 2021 and treated with radical and postoperative radiotherapy. Flow cytometry and the agglutination method were used to determine the proportion of NKT cells in peripheral blood and the TAP coagulation area, respectively before and after radiotherapy. The relationships of clinical features and the cellular features such as changes in NKT cells and ATPs with local recurrence and long-term survival were analyzed. The χ2 test or Fisher′s exact test was employed for intergroup comparison. The Kaplan-Meier method and the Cox model were utilized for univariate and multivariate survival prognosis analyses, respectively. The bivariate Pearson linear correlation analysis was conducted to analyze the relationship between NKT and TAP. Results:The median follow-up time of the whole group was 25 months. Regarding the 1-, 2-, and 3-year survival rates, the local-regional recurrence-free survival (LRRFS) rates were 76.2%, 67.3%, and 64.4%, respectively, the distant metastasis-free survival (DMFS) rates 91.1%, 90.1%, and 89.1%, respectively, and the progression-free survival (PFS) rates 69.3%, 59.4%, and 55.4%, respectively. The 3-year overall survival (OS) rate was influenced by age, surgery, N stage, TNM stage, NKT cell ratio, and TAP, while the 3-year PFS rate was affected by TAP, sex, N stage, and TNM stage. Multivariate analysis suggests that independent adverse prognostic factors for HNSCC included sex, age, N stage, NKT cells, and TAP ( HR=3.00, 2.35, 2.27, 2.02, 2.56, P<0.05). The correlation analysis indicates a positive correlation between NKT cells and TAP ( r=0.26, P=0.009). Conclusions:Stage Ⅲ-Ⅳ B HNSCC treated with radical and postoperative radiotherapy is subjected to a high recurrence rate. Further research is required for the expression levels of NKT cells and TAP in peripheral blood, as well as the influence of their changes during radiotherapy on the 3-year OS, PFS, and LRRFS rates of locally advanced HNSCC.

Objective:To explore the clinical value of Ki-67 and human epidermal growth factor receptor 2 (HER-2) in salivary duct carcinoma in stage Ⅲ-Ⅳ A. Methods:The data of 52 cases of locally advanced salivary duct carcinoma(SDC) diagnosed from January 2012 to December 2020 were retrospectively analyzed. All patients underwent radical surgery and postoperative radiotherapy. Among them, 15.4% of patients had local recurrence, 28.8% had distant metastasis, 17.3% had regional recurrence with distant metastasis. The relationship between clinical features, pathological features such as Ki-67 and HER-2 and prognosis such as local recurrence and distant metastasis was analyzed.Results:The average follow-up time was 37.6 months. The 1- and 2-year local recurrence free survival, distant metastasis free survival, progression free survival were 86.5%, 73.1%, 65.4% and 67.3%, 55.8%, 46.2% respectively. The 3-year progression free survival rate was 33.3%. Comparison between groups showed that age ≥ 65 years old, T stage, TNM stage, vascular tumor thrombus, radiotherapy dose <60 Gy, Ki-67 positive index and HER-2 positive were related to the prognosis of different stages. In multivariate analysis, only age, Ki-67 positive index ≥ 60% and HER-2 protein (3+ ) were independent poor prognostic factors for locally advanced SDC ( t =5.16, 9.84, 8.23, P<0.05). Conclusions:In stage Ⅲ-Ⅳ A SDC, only radical surgery and postoperative radiotherapy have a high rate of distant metastasis. Ki-67 positive index and HER-2 positive are independent adverse prognostic factors.