1.Effect of Salvianolic Acid B Combined with Puerarin on Pyroptosis of SH-SY5Y Cells Injured by Oxygen-glucose Deprivation/Reoxygenation
Shuxin TU ; Yan WANG ; Xiaoli HE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):54-63
ObjectiveTo investigate the effect and mechanism of salvianolic acid B combined with puerarin in protecting the SH-SY5Y cells from the damage by oxygen-glucose deprivation/reoxygenation (OGD/R) based on pyroptosis. MethodSH-SY5Y cells were used to establish the model of OGD/R, and cells were classified into the control, OGD/R, 10 μmol·L-1 salvianolic acid B, 100 μmol·L-1 puerarin, 10 μmol·L-1 salvianolic acid B + 100 μmol·L-1 puerarin, and 10 μmol·L-1 NOD-like receptor protein 3 (NLRP3) inhibitor MCC950 groups. Except the control group, other groups were rapidly reoxygenated for 12 h after 6 h OGD for modeling. The cell survival rate was determined by the methyl thiazolyl tetrazolium (MTT) assay. An optical microscope was used to observe the cell morphology. A spectrophotometer was used to determine the content of lactic dehydrogenase (LDH) in culture supernatant. Cell damage was measured by Hoechst/PI staining. The mRNA levels of NLRP3, cysteinyl aspartate specific proteinase-1 (Caspase-1), gasdermin D (GSDMD), apoptosis-associated speck-like protein (ASC), and interleukin-1β (IL-1β) were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). The protein activation of Caspase-1 and NLRP3 was detected by immunofluorescence. Western blot was employed to determine the protein levels of IL-1β, ASC, NLRP3, Caspase-1, and cleaved Caspase-1. ResultCompared with the control group, the OGD/R group showed decreased cell survival rate (P<0.01), damaged cell morphology, increased leakage rate of LDH (P<0.01), up-regulated mRNA levels of NLRP3, Caspase-1, GSDMD, ASC, and IL-1β (P<0.01), and up-regulated protein levels of IL-1β, ASC, NLRP3, Caspase-1, and cleaved Caspase-1 (P<0.01). Compared with the OGD/R group, salvianolic acid B, puerarin, and salvianolic acid B combined with puerarin improved cell survival rate (P<0.01), and the combined treatment group outperformed salvianolic acid B and puerarin used alone (P<0.01). Salvianolic acid B combined with puerarin and MCC950 both improved cell morphology, reduced the leakage of LDH (P<0.01), alleviated cell damage, and down-regulated the mRNA levels of NLRP3, Caspase-1, GSDMD, ASC, and IL-1β (P<0.05, P<0.01) and also the protein levels of IL-1β, ASC, NLRP3, Caspase-1, and cleaved Caspase-1 (P<0.05, P<0.01). ConclusionThe results indicated that salvianolic acid B combined with puerarin can alleviate the OGD/R-induced damage of SH-SY5Y cells by inhibiting pyroptosis.
2.Chinesization of the HEMO-FISS-QoL questionnaire and its reliability and validity
Songpeng SUN ; Shan JIA ; Fangfang XU ; Tianyu LI ; Zhiyun ZHANG ; Qiaorong CAO ; Xinjian LI ; Yao WU ; Weiping WAN ; Bin SHI ; Jianguo WANG ; Hong NI ; Longyu LIANG ; Xingxiao HUO ; Tianqing YANG ; Lei TIAN ; Ying TIAN ; Mei LIN ; Zhanjun WANG ; Yangyang ZHOU ; Hongchuan CHU ; Riyu LIAO ; Kuerban XIEYIDA ; Junhong LONG ; Shuxin ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(1):75-82
Objective:To evaluate the reliability and validity of the Chinese version of HEMO-FISS-QoL(HF-QoL) questionnaire (HF-QoL-C) in the Chinese population with hemorrhoids.Methods:From November 2021 to November 2022, a self-constructed general information questionnaire, HF-QoL-C, and the 36-item short form health survey (SF-36), Goligher classification, and Giordano severity of hemorrhoid symptom questionnaire (GSQ) were used to conduct a questionnaire survey on 760 hemorrhoid patients in the anorectal department of six hospitals. The data was analyzed for reliability and validity using SPSS 21.0 and AMOS 26.0 software.Results:The Cronbach's α coefficient of HF-QoL-C and its dimension ranged from 0.831 to 0.960, and the split coefficient was 0.832-0.915. Four common factors were extracted through principal component exploratory factor analysis. Confirmatory factor analysis indicated acceptable structural validity( χ2/ df=8.152, RSMEA=0.097, CFI=0.881, IFI=0.881, NFI=0.867). HF-QoL-C was correlated with SF36 and GSQ( r=-0.694, 0.501, both P<0.01). There were differences in the total score and dimensional scores of HF-QoL-C between surgical and drug treated patients, different grades of Goligher classification for hemorrhoidal disease, and different ranges of hemorrhoid prolapse (all P<0.001). No ceiling effect was found in the total score and the scores of each dimension(0.3%-2.0%). There was a floor effect in both psychological function and sexual activity dimensions (16.7%, 35.1%). Conclusion:HF-QoL-C has good reliability and validity, which can be used to measure the quality of life of Chinese hemorrhoid patients.
3.Discussion on clinical application of Dazhui acupoint based on data mining
Yaqi YAO ; Xiang ZHANG ; Yinxiang XU ; Shuxin XIONG ; Kaiqi MIAO ; Guang'an WANG
International Journal of Traditional Chinese Medicine 2024;46(2):243-248
Objective:To discuss dominant symptoms and compatibility rules of Dazhui(GV14) based on data mining.Methods:Literature related to Dazhui (GV14) was retrieved from CNKI, Wangfang, VIP, China Biomedical Literature Database (CBM) and Pubmed databases from January 1, 2012 to August 15, 2022, and the main symptoms of Dazhi (GV14) and the compatibility of acupoints were summarized. Gephi 0.9.5 software was used for complex network analysis to compare the treatment for dominant symptoms with single acupoint of Dazhi (GV14) and the compatibility of the acupoint. SPSS Modeler 18.0 software was used to analyze the association rules of acupoint combination based on Apriori algorithm. The clustering analysis of high frequency acupoints was carried out by SPSS Statistics 26.0 software.Results:A total of 722 articles were included, involving 732 prescriptions. The dominant symptoms of single acupoint were cervical spondylosis, acne, and cold; the treatment for dominant symptoms with compatibility included 14 types, such as cervical spondylosis, allergic rhinitis, ischemic stroke sequelae. The meridian compatibility was dominated by bladder meridian, and the frequency of yang meridians was higher than yin meridians. The compatibility of specific acupoints such as Xiahe acupoint, Beishu acupoint and Bahui acupoint were the main acupoints, and the high frequency acupoints were 33 acupoints such as Feishu (BL13), Baihui (GV20), Fengchi (GB20) and Zusanli (ST36), obtaining 4 series and 8 types of compatible combinations of Dazhui (GV14).Conclusions:Dazhui (GV14) is widely used in the treatment of internal diseases, such as respiratory diseases, nervous system diseases and vertebral artery type of cervical spondylosis. It tends to be flexibly used with multiple compatibility and clustering combination of specific acupoints.
4.Effects of Point Ligation Therapy Combined with Acupoint Catgut Embedment on Facial Nerve Electromyo-graphy and Serum Neurotrophic Factors for Patients with Refractory Facial Paralysis
Hongzhu LI ; Guoan LAI ; Shiyu LIN ; Xiuyi MA ; Guangxian CHEN ; Yuemei LI ; Shuxin WANG
Journal of Traditional Chinese Medicine 2024;65(15):1578-1585
ObjectiveTo observe the clinical effectiveness of point ligation therapy combined with catgut embedment in acupoint for patients with refractory facial paralysis, and to explore the possible mechanisms from the perspective of neurotrophic factors. MethodsTotally 168 patients with intractable facial paralysis were randomly divided into point ligation plus catgut embedment group and electroacupuncture group, with 84 cases in each group. Both groups took methylcobalamin tablets orally, on the basis of which the point ligation plus catgut embedment group gave point ligation therapy at Quanliao (SI 18), Dicang (ST 4), Jiache (ST 6), and catgut embedment in the local acupoints on the affected side combined with the distal acupoints, and the point ligation therapy combined with catgut embedment in acupoint was performed once every 30 days for three treatments; the electroacupuncture group took the same acupoints and gave electroacupuncture, with sparse and dense waves, once every other day. Both groups were treated for 90 days. Before treatment and after 30, 60, 90 and 120 days (follow-up), patients of both groups were observed for House-Brackmann Facial Nerve Grading (HB) and modified Portmann score; facial nerve electromyography was performed before treatment and after 90 days of treatment to record the wave amplitude and latency of facial nerve motor conduction, and serum brain-derived nerve growth factor (BDNF), nerve growth factor (NGF), fibroblast growth factor (FGF2) levels were determined. The clinical effectiveness of both groups after 90 days of treatment was compared, and the adverse events occurred in the course of treatment in all patients were recorded. ResultsAt 30 days, 60 days, 90 days and follow-up visits, the HB grading of patients in both groups improved compared with that before treatment (P<0.05); the distribution of the number of patients with HB grading in the point ligation plus catgut embedment group was superior to that in the electroacupuncture group after 60 days and 90 days of treatment and at follow-up visits (P<0.05). The maximal amplitude of the facial nerve electromyography in both groups after 90 days of treatment was large, and the latency was shorter when compared with those before treatment (P<0.05), and the maximum wave amplitude of the point ligation plus catgut embedment group was higher than that of the electro-acupuncture group, and the latency period was shorter than that of the electro-acupuncture group (P<0.05). The modified Portmann scores of the both groups of patients after 30, 60 and 90 days of treatment and follow-up were higher than those before treatment, and the scores of the point ligation plus catgut embedment group were higher than those of the electroacupuncture group (P<0.05). Serum BDNF, NGF and FGF2 levels in both groups after 90 days of treatment were higher than before, and the point ligation plus catgut embedment group was significantly higher than the electroacupuncture group (P<0.05). The total clinical effective rate of the point ligation plus catgut embedment group (92.68%) was higher than that of the electroacupuncture group (81.01%, P<0.05). Adverse reactions occurred in both groups, mainly including local haematoma or subcutaneous bleeding, allergic reaction, severe pain, and needle fainting, all of which disappeared after symptomatic treatment. ConclusionThe point ligation therapy combined with catgut embedment in acupoint for patients with refractory facial paralysis are effective. The therapy improve the facial nerve function, and its mechanism of action may be related to increasing the level of serum neurotrophic factor and thus promoting facial nerve repairment.
5.Expert consensus on surgical treatment of oropharyngeal cancer
China Anti-Cancer Association Head and Neck Oncology Committee ; China Anti-Cancer Association Holistic Integrative Oral Cancer on Preventing and Screen-ing Committee ; Min RUAN ; Nannan HAN ; Changming AN ; Chao CHEN ; Chuanjun CHEN ; Minjun DONG ; Wei HAN ; Jinsong HOU ; Jun HOU ; Zhiquan HUANG ; Chao LI ; Siyi LI ; Bing LIU ; Fayu LIU ; Xiaozhi LV ; Zheng-Hua LV ; Guoxin REN ; Xiaofeng SHAN ; Zhengjun SHANG ; Shuyang SUN ; Tong JI ; Chuanzheng SUN ; Guowen SUN ; Hao TIAN ; Yuanyin WANG ; Yueping WANG ; Shuxin WEN ; Wei WU ; Jinhai YE ; Di YU ; Chunye ZHANG ; Kai ZHANG ; Ming ZHANG ; Sheng ZHANG ; Jiawei ZHENG ; Xuan ZHOU ; Yu ZHOU ; Guopei ZHU ; Ling ZHU ; Susheng MIAO ; Yue HE ; Jugao FANG ; Chenping ZHANG ; Zhiyuan ZHANG
Journal of Prevention and Treatment for Stomatological Diseases 2024;32(11):821-833
With the increasing proportion of human papilloma virus(HPV)infection in the pathogenic factors of oro-pharyngeal cancer,a series of changes have occurred in the surgical treatment.While the treatment mode has been im-proved,there are still many problems,including the inconsistency between diagnosis and treatment modes,the lack of popularization of reconstruction technology,the imperfect post-treatment rehabilitation system,and the lack of effective preventive measures.Especially in terms of treatment mode for early oropharyngeal cancer,there is no unified conclu-sion whether it is surgery alone or radiotherapy alone,and whether robotic minimally invasive surgery has better func-tional protection than radiotherapy.For advanced oropharyngeal cancer,there is greater controversy over the treatment mode.It is still unclear whether to adopt a non-surgical treatment mode of synchronous chemoradiotherapy or induction chemotherapy combined with synchronous chemoradiotherapy,or a treatment mode of surgery combined with postopera-tive chemoradiotherapy.In order to standardize the surgical treatment of oropharyngeal cancer in China and clarify the indications for surgical treatment of oropharyngeal cancer,this expert consensus,based on the characteristics and treat-ment status of oropharyngeal cancer in China and combined with the international latest theories and practices,forms consensus opinions in multiple aspects of preoperative evaluation,surgical indication determination,primary tumor re-section,neck lymph node dissection,postoperative defect repair,postoperative complication management prognosis and follow-up of oropharyngeal cancer patients.The key points include:① Before the treatment of oropharyngeal cancer,the expression of P16 protein should be detected to clarify HPV status;② Perform enhanced magnetic resonance imaging of the maxillofacial region before surgery to evaluate the invasion of oropharyngeal cancer and guide precise surgical resec-tion of oropharyngeal cancer.Evaluating mouth opening and airway status is crucial for surgical approach decisions and postoperative risk prediction;③ For oropharyngeal cancer patients who have to undergo major surgery and cannot eat for one to two months,it is recommended to undergo percutaneous endoscopic gastrostomy before surgery to effectively improve their nutritional intake during treatment;④ Early-stage oropharyngeal cancer patients may opt for either sur-gery alone or radiation therapy alone.For intermediate and advanced stages,HPV-related oropharyngeal cancer general-ly prioritizes radiation therapy,with concurrent chemotherapy considered based on tumor staging.Surgical treatment is recommended as the first choice for HPV unrelated oropharyngeal squamous cell carcinoma(including primary and re-current)and recurrent HPV related oropharyngeal squamous cell carcinoma after radiotherapy and chemotherapy;⑤ For primary exogenous T1-2 oropharyngeal cancer,direct surgery through the oral approach or da Vinci robotic sur-gery is preferred.For T3-4 patients with advanced oropharyngeal cancer,it is recommended to use temporary mandibu-lectomy approach and lateral pharyngotomy approach for surgery as appropriate;⑥ For cT1-2N0 oropharyngeal cancer patients with tumor invasion depth>3 mm and cT3-4N0 HPV unrelated oropharyngeal cancer patients,selective neck dissection of levels ⅠB to Ⅳ is recommended.For cN+HPV unrelated oropharyngeal cancer patients,therapeutic neck dissection in regions Ⅰ-Ⅴ is advised;⑦ If PET-CT scan at 12 or more weeks after completion of radiation shows intense FDG uptake in any node,or imaging suggests continuous enlargement of lymph nodes,the patient should undergo neck dissection;⑧ For patients with suspected extracapsular invasion preoperatively,lymph node dissection should include removal of surrounding muscle and adipose connective tissue;⑨ The reconstruction of oropharyngeal cancer defects should follow the principle of reconstruction steps,with priority given to adjacent flaps,followed by distal pedicled flaps,and finally free flaps.The anterolateral thigh flap with abundant tissue can be used as the preferred flap for large-scale postoperative defects.
6.Effect of total flavones of Dracocephalum moldavica L.on high-glucose in-duced oxidative damage to retinal ganglion cells and its mechanism
Liying GU ; Shengfu YANG ; Qiming ZHANG ; Shuxin WANG
Recent Advances in Ophthalmology 2024;44(12):937-942
Objective To investigate the effects of total flavones of Dracocephalum moldavica L.(TFDM)on oxida-tive damage to retinal ganglion cells(RGCs)induced by high glucose(HG)and its mechanism.Methods RGCs of mice were taken as the research subjects.RGCs were inoculated in 24-well plates(with 2.5 × 104 cells in each well)and divided into control group(cultured with medium containing 10%fetal bovine serum for 48 h),HG group(cultured with medium containing 30 mmol·L-1 glucose for 48 h),HG+TFDM-L group(cultured with medium containing 30 mmol·L-1 glucose and 25 mg·L-1 TFDM for 48 h),HG+TFDM-M group(cultured with medium containing 30 mmol·L-1 glucose and 50 mg·L-1 TFDM for 48 h),HG+TFDM-H group(cultured with medium containing 30 mmol·L-1 glucose and 100 mg·L-1 TFDM for 48 h),miR-NC group(transfected with miR-NC and then cultured with medium containing 30 mmol·L-1 glu-cose for 48 h),miR-93-5p group(transfected with miR-93-5p mimics and then cultured with medium containing 30 mmol·L-1 glucose for 48 h),anti-miR-NC group(transfected with anti-miR-NC and then cultured with medium containing 100 mg·L-1 TFDM and 30 mmol·L-1 glucose for 48 h),and anti-miR-93-5p group(transfected with anti-miR-93-5p and then cultured with medium containing 100 mg·L-1 TFDM and 30 mmol·L-1 glucose for 48 h).Levels of RGCs oxidative stress indexes in each group were detected according to the kit instructions.The thiobarbituric acid method was used to measure the level of malondialdehyde(MDA),the colorimetric method was adopted to detect the levels of catalase(CAT)and 8-hydroxydeoxyguanosine(8-OHdG),apoptosis rate was detected by flow cytometry,the messenger ribonucleic acid(mR-NA)expressions were analyzed by real-time quantitative polymerase chain reaction,the targeting regulation of miR-93-5p and E2F transcription factor 1(E2F1)were verified by dual luciferase reporter assay,and the protein expressions were de-tected by Western blot.Statistical analysis was conducted on the data of each group.Results In the HG group,the lev-els of MDA and 8-OHdG,apoptosis rate and Bax protein expression of RGCs were higher than those in the control group(all P<0.05);the CAT level and Bcl-2 protein expression were lower than those in the control group(both P<0.05).In the HG+TFDM-L group,HG+TFDM-M group and HG+TFDM-H group,the levels of MDA and 8-OHdG,apoptosis rate and Bax protein expression of RGCs were lower than those in the HG group(all P<0.05);the CAT level and Bcl-2 protein expression were higher than those in the HG group,and those indexes gradually tended to those in the control group with the increase of TFDM concentrations(all P<0.05).In the HG group,the miR-93-5p expression in RGCs was lower than that in the control group,and the mRNA and protein expressions of E2F1 were higher than those in the control group(all P<0.05).In the HG+TFDM-L group,HG+TFDM-M group and HG+TFDM-H group,the miR-93-5p expression in RGCs was higher than that in the HG group,and the mRNA and protein expressions of E2F1 were lower than those in the HG group(all P<0.05).The protein expression of E2F1 in RGCs of the miR-93-5p group was lower than that in the miR-NC group,and the protein expression of E2F1 in RGCs of the anti-miR-93-5p group was higher than that in the anti-miR-NC group(both P<0.05).In the miR-93-5p group,the miR-93-5p expression was higher than that in the miR-NC group,the levels of MDA and 8-OHdG,apoptosis rate,and protein expressions of Bax and E2F1 were lower than those in the miR-NC group,and the CAT level and Bcl-2 protein expression were higher than those in the miR-NC group(all P<0.05).Con-clusion TFDM can inhibit oxidative stress and cell apoptosis,and then reduce the damage to RGCs induced by HG.The mechanism may involve the regulation of miR-93-5p/E2Fl expression.
7.Correlation analysis between AIP and vulnerable plaques of coronary CT
Xuankai ZHOU ; Xiaowen HAN ; Guan WANG ; Zhi ZHU ; Shuxin ZHAO ; Fan CUI ; Jiahang SHI ; Chenghao PIAO
China Medical Equipment 2024;21(10):46-50
Objective:To explore the relationship between atherosclerotic index of plasma(AIP)and vulnerable plaque of coronary under computed tomography(CT)based on coronary CT angiography(CCTA).Methods:Data were retrospectively collected on 213 patients with coronary heart disease(CHD)who underwent CCTA examination from January 2021 to February 2024 at the Second Affiliated Hospital of Shenyang Medical College,and they were divided into a vulnerable plaque group(123 cases)and a non-vulnerable plaque group(90 cases)according to whether existed vulnerable plaque of coronary artery.General clinical data such as age,gender,history of smoking,history of alcohol consumption,history of diabetes,and serum indicators such as AIP were collected.The differences in AIP and other factors between the two groups were compared.The independent influencing factors of vulnerable plaque of coronary artery were determined by multifactorial logistic regression analysis,and the predictive value of AIP for vulnerable plaque was assessed by drawing a receiver operating characteristic(ROC)curve.Results:AIP of vulnerable plaque group was 0.22±0.31,which was higher than that 0.05±0.27 of the vulnerable plaque group,and the difference of AIP between two groups was significant(t=4.223,P<0.001).Multifactorial logistic analysis showed there was independent correlation between AIP and vulnerable plaque under CT(OR=7.556,95%CI:2.442~23.385,P=0.002).The ROC curve showed that the best cut-off value of AIP was 0.20 in predicting vulnerable plaque under CT,and the value of area under curve(AUC)was 0.665,and the sensitivity was 55.56%and the specificity was 73.98%.Conclusion:AIP is an independent influencing factor for CHD patients who complicate vulnerable plaques,and it has a certain of predictive value for vulnerable plaques.
8.Meta-analysis of comparison of efficacies between low-dose and standard-dose radiotherapy in human papillomavirus-associated oropharyngeal cancer
Jiewen HAO ; Shuxin WEN ; Li ZHANG ; Chen WANG ; Shuntu CHE ; Junlong LI ; Meifang LUO
Chinese Journal of Radiation Oncology 2024;33(7):599-605
Objective:To compare the efficacy and safety of low-dose and standard-dose radiotherapy (LD-RT vs. SD-RT) in the treatment of oropharyngeal cancer patients with positive human papillomavirus (HPV). Methods:All comparative studies of low-dose versus standard-dose radiotherapy in the treatment of HPV-positive oropharyngeal cancer were searched from PubMed, Web of Science, Cochrane Library, EMbase, Chinese Biomedical Literature Database (CBM), CNKI, Chongqing VIP, and Wanfang databases from January 1, 2000 to February 9, 2023. According to the inclusion and exclusion criteria, the data were strictly screened, and the RevMan 5.4 software was used for meta-analysis.Results:A total of 8 studies were included. The pooled results showed that the overall survival (OS) and progression-free survival (PFS) of patients in the LD-RT group were similar to those in the SD-RT group ( HR=0.83, 95% CI=0.59-1.18, P=0.31; HR=0.97, 95% CI=0.53-1.78, P=0.92), but the rate of percutaneous endoscopic gastrostomy (PEG) insertion was significantly reduced ( RR=0.45, 95% CI=0.28-0.72, P=0.001). Conclusion:LD-RT yields similar efficacy and lower rate of PEG insertion compared with SD-RT in HPV-positive oropharyngeal neoplasm patients.
9.Construction and validation of a prognostic model for colon cancer based on inflammatory response-related genes
Tao ZHANG ; Shiying LI ; Tao JING ; Zihao LIU ; Shuangshuang JI ; Mingxing LIU ; Huiru JI ; Lihong WANG ; Shuxin ZHANG
Cancer Research and Clinic 2023;35(5):353-360
Objective:To screen the differentially expressed genes (DEG) related to inflammatory response associated with the prognosis of colon cancer based on the bioinformatics approach, and to construct and validate a prognostic model for colon cancer.Methods:RNA sequencing and clinical data of 472 colon cancer patients and normal colon tissues of 41 healthy people were retrieved from the Cancer Genome Atlas (TCGA) database. Gene expression related to prognosis of colon cancer and clinical data were retrieved from the International Cancer Genome Consortium (ICGC) database. The retrieval time was all from the establishment of library to November 2022. A total of 200 genes associated with inflammatory response obtained from the Gene Set Enrichment Analysis (GSEA) database were compared with the RNA sequencing gene dataset of colon cancer and normal colon tissues obtained from the TCGA database, and then DEG associated with inflammatory response were obtained. The prognosis-related DEG in the TCGA database were analyzed by using Cox proportional risk model, and the inflammatory response-related DEG were intersected with the prognosis-related DEG to obtain the prognosis-related inflammatory response-related DEG. The prognostic model of colon cancer was constructed by using LASSO Cox regression. Risk scores were calculated, and colon cancer patients in the TCGA database were divided into two groups of low risk (< the median value) and high risk (≥the median value) according to the median value of risk scores. Principal component analysis (PCA) was performed on patients in both groups, and survival analysis was performed by using Kaplan-Meier method. The efficacy of risk score in predicting the overall survival (OS) of colon cancer patients in the TCGA database was analyzed based on the R software timeROC program package. Clinical data from the ICGC database were applied to externally validate the constructed prognostic model, and patients with colon cancer in the ICGC database were classified into high and low risk groups based on the median risk score of patients with colon cancer in the TCGA database. By using R software, single-sample gene set enrichment analysis (ssGESA), immunophenotyping difference analysis, immune microenvironment correlation analysis, and immune checkpoint gene difference analysis of immune cells and immune function were performed for prognosis-related inflammation response-related DEG in the TCGA database.Results:A total of 60 inflammatory response-related DEG and 12 prognosis-related DEG were obtained; and 6 prognosis-related inflammatory response-related DEG (CCL24, GP1BA, SLC4A4, SRI, SPHK1, TIMP1) were obtained by taking the intersection set. LASSO Cox regression analysis showed that a prognostic model for colon cancer was constructed based on 6 prognosis-related inflammatory response-related DEG, and the risk score was calculated as = -0.113×CCL24+0.568×GP1BA+ (-0.375)×SLC4A4+(-0.051)×SRI+0.287×SPHK1+0.345×TIMP1. PCA results showed that patients with colon cancer could be better classified into 2 clusters. The OS in the high-risk group was worse than that in the low-risk group in the TCGA database ( P < 0.001); the area of the curve (AUC) of the prognostic risk score for predicting the OS rates of 1-year, 3-year, 5-year was 0.701, 0.685, and 0.675, respectively. The OS of the low-risk group was better than that of the high-risk group in the ICGC database; AUC of the prognostic risk score for predicting the OS rates of 1-year, 2-year, 3-year was 0.760, 0.788, and 0.743, respectively. ssGSEA analysis showed that the level of immune cell infiltration in the high-risk group in the TCGA database was high, especially the scores of activated dendritic cells, macrophages, neutrophils, plasmacytoid dendritic cells, T helper cells, and follicular helper T cells in the high-risk group were higher than those in the low-risk group, while the score of helper T cells 2 (Th2) in the high-risk group was lower compared with that in the low-risk group (all P < 0.05); in terms of immune function, the high-risk group had higher scores of antigen-presenting cell (APC) co-inhibition, APC co-stimulation, immune checkpoint, human leukocyte antigen (HLA), promotion of inflammation, parainflammation, T-cell stimulation, type Ⅰ interferon (IFN) response, and type ⅡIFN response scores compared with those in the low-risk group (all P < 0.05). The results of immunophenotyping analysis showed that IFN-γ-dominant type (C2) had the highest inflammatory response score, and the differences were statistically significant when compared with trauma healing type (C1) and inflammatory response type (C3), respectively (all P < 0.05). Immune microenvironment stromal cells and immune cells were all positively correlated with prognostic risk scores ( r values were 0.35 and 0.21, respectively, both P < 0.01). The results of immune checkpoint difference analysis showed there was a statistically significant difference in programmed-death receptor ligand 1 (PD-L1) expression level between high-risk group and low-risk group ( P = 0.002), and PD-L1 expression level was positively correlated with prognostic risk score ( r = 0.23, P < 0.01). Conclusions:Inflammatory response-related genes may play an important role in tumor immunity of colon cancer and can be used in the prognostic analysis and immunotherapy of colon cancer patients.
10.Removal of anti-phospholipase A2 receptor autoantibodies in primary membranous nephropathy by recombinant phospholipase A2 receptor tandem epitope immunosorbent
Kexin WANG ; Shuxin LIU ; Fangling JI ; Jiahang QIAN ; Jun REN ; Lulu HAN ; Lingyun JIA
Chinese Journal of Nephrology 2023;39(11):840-845
Objective:To investigate the role of recombinant phospholipase A2 receptor (PLA2R) tandem dominant epitopes (PLA2RTD) in the removal of anti-PLA2R autoantibodies (anti-PLA2R) from primary membranous nephropathy (PMN).Methods:The recombinant protein PLA2RTD (cysteine-rich domain, C-type lectin like domain 1 and C-type lectin like domain 7) was expressed in bacmid-insect cell expression system. Circular dichroism was used to determine the secondary structure of PLA2RTD. Enzyme-linked immunosorbent assay and immunofluorescence were used to determine the biological activity of PLA2RTD. Epoxy activation method was used to couple the recombinant PLA2RTD and agarose gel CL-6B microspheres for preparing specific immune adsorbent of anti-PLA2R.Results:The study achieved the expression of PLA2RTD in the first time from the bacmid-insect cell system, demonstrating the good immunogenicity and high binding specificity of PLA2RTD. A single in vitro adsorption of PLA2RTD could averagely eliminate 76.66% of anti-PLA2R [(6.66±0.30) RU/ml vs. (28.54±2.10) RU/ml], the changes of IgG, IgA, albumin, β2 microglobulin, interleukin 6, and tumor necrosis factor α were all less than 4% after completion of adsorption, and the second or third repeated use of PLA2RTD could maintain the adsorption efficiency of about 65%. Conclusion:PLA2RTD-based specific immunosorbent can effectively remove anti-PLA2R in plasma, which provides a new way to specifically remove PMN-related autoantibodies.


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