1.Protective effect of sericin on streptozotocin-induced INS-1 cell damage by regulating PI3K/Akt/NF-κB signaling pathway through Akt1 and its mechanism
Cheng CHEN ; Jingyao LI ; Wanxiang HU ; Donghui LIU ; Zhihong CHEN
Journal of Jilin University(Medicine Edition) 2025;51(3):590-598
Objective:To discuss the effect of sericin on the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/nuclear factor-κB(NF-κB)signaling pathway and apoptosis in the streptozotocin(STZ)-damaged INS-1 cells,and to clarify its mechanism.Methods:The INS-1 cells were cultured with complete medium containing 0,0.1,0.3,1.0,3.0,and 10.0 μmol·L-1 Akt1 inhibitor A-674563,10 mmol·L-1 STZ,and 600 mg·L-1 sericin,and divided into 0,0.1,0.3,1.0,3.0,and 10.0 μmol·L-1 A-674563 groups,and the control group(complete medium without drugs)was set up.Cell counting kit-8(CCK-8)method was used to detect the survival rates of the INS-1 cells,and the half-maximal inhibitory concentration(IC50)value was calculated to determine the optimal inhibitory concentration of A-674563,which was further verified by Western blotting method.The INS-1 cells were divided into normal control group(complete medium),model group(10 mmol·L-1 STZ+complete medium),and low,medium,and high doses of sericin groups(10 mmol·L-1 STZ+150 mg·L-1 sericin+complete medium,10 mmol·L-1 STZ+300 mg·L-1 sericin+complete medium,and 10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium).CCK-8 method was used to detect the survival rates of the INS-1 cells in various groups to determine the optimal concentration of sericin.Additionally,the INS-1 cells were divided into normal control group(complete medium),model group(10 mmol·L-1 STZ+complete medium),sericin group(10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium),and A-674563 group(10 mmol·L-1 STZ+600 mg·L-1 sericin+0.3 μmol·L-1 A-674563+complete medium).Flow cytometry was used to detect the apoptotic rates of the INS-1 cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Akt1,NF-κB,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)mRNA in the INS-1 cells in various groups;Western blotting method was used to detect the expression levels of phosphorylated Akt1(p-Akt1)and NF-κB proteins in the INS-1 cells in various groups;enzyme linked immunosorbent assay(ELISA)method was used to detect the levels of TNF-α and IL-6 in the INS-1 cells in various groups.Results:The survival rates of the INS-1 cells in control group was 100.00%±0.00%;in 0,0.1,0.3,1.0,3.0,and 10.0 μmol·L-1 A-674563+10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium groups,which were 82.50%±2.28%,69.47%±1.94%,51.51%±1.74%,38.94%±1.57%,24.79%±1.14%,and 19.85%±1.03%,respectively.The IC?? value of A-674563 for INS-1 cells was 0.3 μmol·L-1,and 0.3 μmol·L-1 A-674563 was selected for subsequent experiments.Compared with 0 μmol·L-1 A-674563,the expression level of p-Akt1 protein in the INS-1 cells after treated with 0.3 μmol·L-1 A-674563+10 mmol·L-1 STZ+600 mg·L-1 sericin+complete medium was significantly decreased(P<0.05).The CCK-8 results showed that compared with normal control group,the survival rate of the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the survival rates of the INS-1 cells in low,medium,and high doses of sericin groups were significantly increased(P<0.05);compared with low and medium doses of sericin groups,the survival rate of the INS-1 cells in high dose of sericin group was significantly increased(P<0.05).Thus,600 mg·L-1 sericin was selected for subsequent experiments.The CCK-8 results showed that compared with normal control group,the survival rate of the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the survival rate of the INS-1 cells in sericin group was significantly increased(P<0.05);compared with sericin group,the survival rate of the INS-1 cells in A-674563 group was significantly decreased(P<0.05).The flow cytometry results showed that compared with normal control group,the apoptotic rate of the INS-1 cells in model group was significantly increased(P<0.05);compared with model group,the apoptotic rate of the INS-1 cells in sericin group was significantly decreased(P<0.05);compared with sericin group,the apoptotic rate of the INS-1 cells in A-674563 group was significantly increased(P<0.05).The RT-qPCR results showed that compared with normal control group,the expression level of Akt1 mRNA in the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the expression levels of Akt1 mRNA in the INS-1 cells in low,medium,and high doses of sericin groups were significantly increased(P<0.05);compared with low and medium doses of sericin groups,the expression level of Akt1 mRNA in the INS-1 cells in high dose of sericin group was significantly increased(P<0.05).Compared with normal control group,the expression levels of NF-κB,TNF-α,and IL-6 mRNA in the INS-1 cells in model group were significantly increased(P<0.05);compared with model group,the expression levels of NF-κB,TNF-α,and IL-6 mRNA in the INS-1 cells in sericin group were significantly decreased(P<0.05);compared with sericin group,the expression level of NF-κB mRNA in the INS-1 cells in A-674563 group was significantly increased(P<0.05).The Western blotting results showed that compared with normal control group,the expression level of p-Akt1 protein in the INS-1 cells in model group was significantly decreased(P<0.05);compared with model group,the expression levels of p-Akt1 protein in the INS-1 cells in low,medium,and high doses of sericin groups were significantly increased(P<0.05);compared with low and medium doses of sericin groups,the expression level of p-Akt1 protein in the INS-1 cells in high dose of sericin group was significantly increased(P<0.05).Compared with normal control group,the expression level of NF-κB protein in the INS-1 cells in model group was significantly increased(P<0.05);compared with model group,the expression level of NF-κB protein in the INS-1 cells in sericin group was significantly decreased(P<0.05);compared with sericin group,the expression level of NF-κB protein in the INS-1 cells in A-674563 group was significantly increased(P<0.05).The ELISA results showed that compared with normal control group,the levels of TNF-α and IL-6 in the INS-1 cells in model group were significantly increased(P<0.05);compared with model group,the levels of TNF-α and IL-6 in the INS-1 cells in sericin group were significantly decreased(P<0.05);compared with sericin group,the levels of TNF-α and IL-6 in the INS-1 cells in A-674563 group were significantly increased(P<0.05).Conclusion:Sericin alleviates the PI3K/Akt/NF-κB signaling pathway-mediated inflammatory response and apoptosis by targeting Akt1,exerting a protective effect against STZ-induced damage in INS-1 cells.
2.Analysis of clinical study registration characteristics of periodontitis based on ClinicalTrials.gov and ChiCTR databases
Jiacheng DAI ; Cong LI ; Liye QIN ; Guihua YE ; Ziyu YE ; Wanxiang YE ; Jincheng ZENG
Chongqing Medicine 2025;54(7):1597-1603
Objective To extract and summarize the clinical registration information of periodontitis registered in the US ClinicalTrials.gov and Chinese Clinical Trial Registry(ChiCTR),and further analyze the registration characteristics of periodontitis clinical trials.Methods The ClinicalTrials.gov and ChiCTR data-bases were searched and compiled for periodontitis clinical registration information from 2000 to December 26,2024,including registration number,country/region of registration,annual number of registered projects,sample size,study type and design,study phase,and trial progress status.Results As of December 26,2024,a total of 520 242 registered clinical trials were retrieved from the ClinicalTrials.gov registry platform,of which 1 542(0.30%)were related to periodontitis.There were 189(12.26%)studies on periodontitis-related pro-jects in Turkey,while a total of 37(2.4%)projects were initiated by researchers in China,which ranked ninth.The Chinese Clinical Trial Register(ChiCTR)had 92 954 registered projects,of which 165 were on pe-riodontitis,and most of them were conducted by well-known affiliated hospitals and stomatology hospitals.The number of registrations in the ClinicalTrials.gov database increased year by year and reached a peak in 2022(146 registrations).Trial designs were focused on interventional and observational studies.ClinicalTri-als.gov study phases were focused on phases 2 and 4,while ChiCTR was clustered at phase 0(pre-registra-tion).Conclusion The number of clinical registrations for periodontitis in China's database is relatively low,and despite a steady upward trend,there is still a gap compared with other countries internationally.Future re-search should focus on how to encourage more oral health related research institutions to register on the plat-form and how to increase the sample size.
3.Research progress on the mechanism of ferroptosis on osteosarcoma
Yize YAN ; Long YUAN ; Wanxiang LI ; Sen LI ; Jichao BIAN ; Guodong WANG
Journal of Chinese Physician 2024;26(1):142-146
Osteosarcoma is the most common malignant bone tumor disease in young children and young people. It usually has strong invasiveness, and conventional treatment cannot achieve the expected results. Therefore, studying the mechanism of tumor cell death and exploring more effective treatment methods is of great significance. As a new form of cell death, ferroptosis has been found to have three main regulatory pathways closely related to tumor cell molecular mechanisms, genes, etc. This provides a theoretical basis for the application of ferroptosis in the treatment of osteosarcoma. This article reviews recent research on the interaction between ferroptosis and osteosarcoma in regulating molecules, genes, and other factors, as well as the application of ferroptosis in the treatment of osteosarcoma.
4.Discovery of a potent and selective cell division cycle 7 inhibitor from 6-(3-fluoropyridin-4-yl)thieno3,2-dpyrimidin-4(3H)-one derivatives as an orally active antitumor agent.
Mingwei FU ; Min GE ; Wanxiang YANG ; Chunchen HU ; Xiaowei LI ; Yuanjiang WANG ; Shaohua GOU
Acta Pharmaceutica Sinica B 2024;14(2):893-896
5.Research progress on the relationship between exosomes and pain
Long YUAN ; Sen LI ; Wanxiang LI ; Jichao BIAN ; Guodong WANG
Journal of Chinese Physician 2022;24(12):1901-1904
Exosomes are extracellular microbubbles related to intercellular communication, which have the ability to transport and transfer biological macromolecules. Pain will lead to a sharp decline in the quality of life of patients, and will give patients and society a heavy medical burden. More and more evidences show that the exosomes plays an important role in the pathological process of pain related diseases.Summarizing the exosomes and the biomolecules carried by them under different pain conditions, and identifying the specific exosomes related to the pain state will be helpful for early diagnosis and treatment of pain related diseases. In addition, the ability of exosomes to transmit information and its widespread characteristics in the body indicate that they have broad prospects as a new diagnosis and treatment tool in the field of pain. A better understanding of the relationship between the exosomes and pain will provide a novel and promising treatment for patients with pain.
6. Individualized prediction of the risk of inguinal lymph node metastasis in patients with squamous cell carcinoma of penile
Guangdong HOU ; Yu ZHENG ; Fuli WANG ; Xi’an LI ; Geng ZHANG ; Longlong ZHANG ; Wanxiang ZHENG ; Jianlin YUAN
Chinese Journal of Urology 2019;40(12):909-914
Objective:
To investigate the independent predictors of inguinal lymph node metastasis (LLM) in patients with penile squamous cell carcinoma (SCCP), and to establish a nomogram for predicting individual LLM risk.
Methods:
The data of patients with SCCP diagnosed at the department of urology, Xijing Hospital from July 2009 to June 2019 were analyzed retrospectively. A total of 101 patients were included in this study, with age of 55 (26-84) years. There were 25 (24.8%) and 76 (75.2%) patients with and without palpable inguinal lymph node enlargement, respectively. There were 47 cases (46.5%), 40 cases (39.6%) and 14 cases (13.9%) in T1, T2 and T3 stages, respectively; there were 67 cases (66.3%), 21 cases (20.8%) and 13 cases (12.9%) in Broder 1, 2 and 3, respectively. The average value (or median) of fibrinogen was 2.84 (1.72-5.00)g/L; alkaline phosphatase was 80(32±214)U, hemoglobin was 147(81-180)g, platelet count was (193.74±65.68×109/L, absolute value of neutrophils, monocytes and lymphocytes were 3.98(1.19-11.85)×109/L, 0.44(0.17-1.90)×109/L and 1.68(0.58-4.13)×109/L, respectively. The average (or median) value of PLR, NLR and LMR were 113.38(18.80-418.42), 2.42(0.59-10.22) and 3.84 (1.08-9.89), respectively. There were 26 cases (25.7%) with LLM and 75 cases (74.3%) without LLM. The independent predictors of LLM were identified by univariate and multivariate logistic regression analyses. The R software was used to establish the nomogram by integrating all independent predictors, and the bootstrap method was used to internally validated our nomogram, where the value of AUC (area under the ROC curve) was calculated and the calibration plot was drawn.
Results:
Clinical inguinal lymph node status (
7.Structural analysis of atypical class 1 integrons in clinical Proteus isolates
Quhao WEI ; Linlin XIAO ; Qingfeng HU ; Wanxiang LI ; Weiwei LIU
Chinese Journal of Microbiology and Immunology 2018;38(6):440-445
Objective To analyze the structures of atypical class 1 integrons in clinical Proteus isolates. Methods This study included 26 class 1 integron integrase gene-positive clinical Proteus isolates, from which the variable regions of class 1 integrons could not be amplified. Six isolates were chosen from them to amplify the flanking DNA segments of class 1 integron integrase gene using inverse PCR. The se-quences of PCR products were analyzed with BLAST to identify the target homologous sequences as well as their accession numbers in GenBank. Primers for overlap PCR were designed according to the flanking se-quences. Then the 26 clinical Proteus isolates were analyzed with overlap PCR and sequencing analysis. Re-sults The variable regions of class 1 integrons in 25 out of the 26 clinical Proteus isolates were completely or partly identified by using inverse PCR,overlap PCR and sequencing analysis. A gene cassette array of estX-psp-aadA2-cmlA1-aadA1a-qacI-tnpA-sul3 was detected in 22 isolates. The variable regions of class 1 integrons in the other three isolates were identified to be estX-psp-aadA2-cmlA1,dfrA14 and IS26,respectively. All of the 25 isolates lacked the 3'conserved segements in class 1 integron. Conclusion Inverse PCR can be used to analyze the structures of atypical class 1 integrons. Gene cassette psp and gene cassette arrays of estX-psp-aadA2-cmlA1-aadA1a-qacI-tnpA-sul3 and estX-psp-aadA2-cmlA1 in clinical Proteus isolates are reported for the first time.
8.Multicenter study on risk factors of peripherally inserted central catheter-related infections among neonates
Zhenzhu LIN ; Xiaoyun XIONG ; Shaoling HE ; Yuanhong LYU ; Li JIN ; Li SUN ; Xiaoyan FANG ; Wanxiang HE
Chinese Journal of Modern Nursing 2018;24(28):3371-3376
Objective To explore the situation and risk factors of peripherally inserted central catheter (PICC)-related infections among neonate so as to provide a nursing reference for preventing catheter-related infections. Methods From September 2015 to June 2017, a prospective study was carried out to 811 neonates with PICC from 7 hospitals in Shenzhen City to observe the incidence of catheter-related infections. Simple correlation and multiple factors unconditional Logistic regression was used to analyze the correlative factors of catheter-related infections. Results Among 811 neonates, there were 770 (94.9%) without and 41 (5.1%, 1.95/1 000 catheter days) with catheter-related infections along with 20 cases with exit-site infection and 21 cases with catheter related bloodstream infections (CRBSI). Top three pathogens of CRBSI included Staphylococcus epidermidis, fungus and klebsiella pneumonia. Simple correlation showed that there were statistical differences in gestational age, birth weight of the neonate, disinfection methods of infusion connector, sterile protective barrier during maintenance of catheter between infection group and non-infection group (χ2=4.026,4.964, 4.369,7.463;P< 0.05). Multiple factors unconditional Logistic regression revealed that the risk factor contained birth weight< 1 200 g (OR=2.099, 95%CI: 1.103-3.996, P< 0.05), and the protective factors consisted of sterile protective barrier during maintenance of catheter (OR=0.393, 95%CI: 0.206-0.749,P<0.01). Conclusions Birth weight <1 200 g, sterile protection during maintenance of catheter are the influencing factors of neonatal PICC catheter-related infections. Sterile protective barrier during maintenance of PICC for neonate should include wearing sterile mask, round hat, using aseptic packets and wearing sterile gloving to maintain the catheter. Aseptic technique should be paid more attention to during indwelling catheter and maintaining catheter for premature with birth weight <1 200 g.
9.Clinical features and treatment of residual gallbladder lesions after cholecystectomy
Yajun GENG ; Shudong LI ; Xingkai MENG ; Chengwang YANG ; Zefeng WANG ; Tao HE ; Wanxiang WANG ; Zhenxia WANG ; Jianjun REN
Chinese Journal of Digestive Surgery 2017;16(4):375-379
Objective To investigate the clinical features and treatment of residual gallbladder lesions after cholecystectomy.Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 83 patients with residual gallbladder lesions after cholecystectomy who were admitted to the Affiliated Hospital of Inner Mongolia Medical University between January 2009 and April 2016 were collected.Among the 83 patients,74 had residual gallbladder (41 combined with bile duct stones and 33 with simple residual gallbladder)and 9 had simple residual bile duct stones.Patients received laboratory and imaging examinations,and then selected suitable surgery according to residual lesions.Observation indicators included:(1) clinical features:medical history,clinical manifestation,features of laboratory and imaging examinations;(2) surgical and postoperative situations:surgical procedure,operation time,volume of intraoperative blood loss,time of drainagetube removal,postoperative complications and duration of postoperative hospital stay;(3) postoperative pathological examination;(4) follow-up situation.Follow-up using outpatient examination and telephone interview was performed to detect survival of patients and recurrence of lesions using abdomen color Doppler ultrasound up to October 2016.Measurement data with normal distribution were represented as ~ ±s.Results (1) Clinical features:① medical history:all the 83 patients had histories of cholecystectomy,including 57 with emergency operation and 26 with selective operation.② Clinical manifestation:initial clinical manifestation occurred at 1-324 months postoperatively,with an average time of 96 months.Of 83 patients,49 had right upper abdominal colic associated with right shoulder or back rediating pain,fever,nausea and vomiting,24 had jaundice as the main symptoms,8 had gradually worse conditions,with right upper quadrant abdominal pain,chills and fever associated with jaundice in the advanced stage and 2 had abdominal discomfort associated with incomplete intestinal obstruction.③ Laboratory examination:elevated alanine transaminase (ALT) and aspartate transaminase (AST) levels were detected in 48 patients,elevated total bilirubin (TBil) level in 28 patients and elevated serum and urinary amylase level in 4 patients,respectively.④ Imaging examination:of 83 patients,77 had positive results of magnetic resonanced eholangio-pancreatography (MRCP),43 had positive results of abdominal color Doppler ultrasound and 39 (4 combined with pancreatitis) had positive results of abdominal computed tomography (CT).(2) Surgical and postoperative situations:all the 83 patients underwent surgery.Of 74 patients with residual gallbladder,72 underwent residual gallbladder resection (of 41 combined with bile duct stones,35 undergoing residual gallbladder resection + bill duct exploration and removing the stone + T-tube drainage,6 initially undergoing endoscopic sphincterotomy for removing bile duct stones and then residual gallbladder resection and 31 undergoing simple residual gallbladder resection),1 combined with residual gallbladder cancer underwent residual gallbladder resection + common bile duct resection + hepatic duct-jejunum internal drainage and 1 combined with residual gallbladder cancer and common bile duct space-occupying lesion underwent palliative biliary-enterostomy.Nine patients with simple residual bile duct stones underwent endoscopic sphincterotomy for removing bile duct stones.Operation time,volume of intraoperative blood loss,time of drainage-tube removal and duration of postoperative hospital stay were (92±39)minutes,(63±12)mL,(5±4)days and (9±5)days in 74 patients with residual gallbladder,respectively.There was no severe postoperative complication.Operation time,volume of intraoperative blood loss and duration of postoperative hospital stay were (57±33)minutes,(25±9)mL and (5 ± 3) days in 9 patients with simple residual bile duct stones,respectively.No severe complication was occurred in 9 patients.(3) Postoperative pathological examination:the length of residual cystic duct in 74 patients with residual gallbladder was (2.8 ± 1.0) cm,combining with stones.Of 74 patients,simple hyperplasia of residual gallbladder were detected in 54 patients,dysplasia in 14 patients,tumor-like hyperplasia and benign tumor in 4 patients and adenocarcinoma in 2 patients.(4) Follow-up situation:all the 83 patients were followed up for 6-71 months,with a median time of 33 months.Eighty-one patients were healthy survival,without recurrence of lesions.One patient had recurrence of bile duct stones at 34 months postoperatively,and then was cured by choledochojejunostomy,without recurrence up to the end of follow-up.One patient with residual gallbladder cancer complicated with common bile duct space-occupying lesion died at 9 months postoperatively.Conclusions Patients have recurrent right upper abdominal colic associated with right shoulder or back rediating pain,fever and jaundice after cholecystectomy,and lesions of residual gallbladder or bile duct stones should be considered.Reoperation is safe and feasible for patients,with a good clinical outcome.
10.Prevention of traditional Chinese medicine Gubiao Pixie prescription for nosocomial infection in elderly population
Wanxiang ZHANG ; Bochao WANG ; Zhijun LI
Chinese Critical Care Medicine 2017;29(5):469-472
Objective To observe the curative effect of traditional Chinese medicine (TCM) Gubiao Pixie prescription on the prevention of nosocomial infection in elderly patients with susceptible factors.Methods A prospective observational study was conducted. The elderly patients aged over 70 years admitted to the Department of Integrated TCM and Western Medicine of Tianjin First Center Hospital from March 2015 to March 2016 were enrolled. The patients were divided into experiment group and control group by random number table. The patients in control group were given conventional treatment without special intervention; and those in experimental group were given Gubiao Pixie prescription particles (prescription composition:Radix Astragali 30 g,Bran fried Rhizoma Atracylodis 20 g,Raidix Saposhnikoviae 12 g,Radix Scutellariae 10 g,Fructus Tsaoko 6 g) on the basis of conventional treatment. The Gubiao Pixie prescription particles were taken in warm water 300 mL, twice a day in morning and evening respectively, taking half an hour after meals, and were increased or decreased according to the disease condition. The changes in immune function parameters before and after treatment, as well as the incidence of nosocomial infection, the abnormal increase rate of body temperature, white blood cell (WBC), and C-reactive protein (CRP) after 10 days of treatment in the two groups were observed.Results A total of 110 elderly patients with susceptible factors during hospitalization were included. After the exclusion of vomiting, abdominal distension and failure to conform the trial requirements, hospitalization time less than 10 days of patients, a total of 100 patients were enrolled in the analysis finally, with 50 patients in control group and in experimental group respectively. There were no significant differences in immune function parameters including IgA, IgG, IgM before treatment between the two groups. After 10 days of treatment, the immune function parameters showed no significant improvement in control group, and those in experiment group were improved significantly, and IgA (g/L: 1.59±0.32 vs. 1.29±0.29), IgG (g/L: 12.07±2.37 vs. 10.23±1.91), and IgM (g/L: 1.01±0.29 vs. 0.88±0.24) were significantly increased as compared with those of control group (allP < 0.01). Compared with the control group, the incidence of nosocomial infection (20% vs. 38%) and the abnormal increase rate of body temperature (24% vs. 44%), WBC (28% vs. 52%), and CRP (28% vs. 50%) 10 days after treatment in experimental group were significantly decreased (allP < 0.05).Conclusions TCM Gubiao Pixie prescription has a role in enhancing immune function and antibacterial and bactericidal effect. It has certain preventive effect on nosocomial infection in susceptible people.

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