1.Effect of Huangqin Qingre Chubi Capsules-containing Serum on CircRNA_0001543/NF-κB Expression in Co-cultured PBMCs and Human FLSs from Patients with Ankylosing Spondylitis
Yajun QI ; Jian LIU ; Qiao ZHOU ; Yuedi HU ; Xiang DING ; Chengzhi CONG ; Xu LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):87-95
ObjectiveThis study aims to explore the effects of Huangqin Qingre Chubi capsules-containing serum on the expression of CircRNA_0001543/nuclear factor-kappa B (NF-κB) in co-cultured peripheral blood mononuclear cells (PBMCs) and human fibroblast-like synoviocytes (FLSs) from patients with ankylosing spondylitis (AS). MethodsVenous blood was collected from patients with AS to isolate PBMCs. FLSs were co-cultured with AS patients' PBMCs, and FLSs were harvested after co-culture for subsequent experiments. The normal control group consisted of normal FLSs, while the model group comprised co-cultured AS PBMCs and FLSs to simulate AS pathology. The Huangqin Qingre Chubi capsules group involved adding Huangqin Qingre Chubi capsules-containing serum to the co-cultured cells(6.48 g·kg-1). To investigate the effect of HQC-containing serum on the viability of co-cultured cells, and the experiment was divided into the following groups based on the dilution concentration: blank group, 10% HQC group, 20% HQC group, and 30% HQC group.To study the influence of the optimal concentration of HQC-containing serum on cytokine and pathway indicators in each group, the experiment was divided into three groups: normal group, model group, and optimal concentration HQC-containing serum group.For the validation of the transfection efficiency of the CircRNA_0001543 interference plasmid, the experiment was divided into the following groups: blank group, si-NC group (with transfection reagent), si-circ_0001543-1 group (with transfection reagent and interference plasmid No. 1 targeting circ_0001543), si-circ_0001543-2 group (with transfection reagent and interference plasmid No. 2 targeting circ_0001543), and si-circ_0001543-3 group (with transfection reagent and interference plasmid No. 3 targeting circ_0001543).For the validation of the transfection efficiency of the CircRNA_0001543 overexpression plasmid, the experiment was divided into the following groups: blank group, OE-NC group (with transfection reagent), and OE-circ_0001543 group (with transfection reagent and overexpression plasmid targeting circ_0001543).To study the effects of CircRNA_0001543 interference/overexpression on cytokine and pathway indicators in each group, the experiment was divided into the following groups: si-NC group, si-CircRNA_0001543 group, OE-NC group, and OE-CircRNA_0001543 group. Enzyme-linked immunosorbent assay (ELISA) was used to detect levels of interleukin-1β (IL-1β), IL-10, IL-37, and tumor necrosis factor-α (TNF-α). Real-time quantitative polymerase chain reaction (Real-time PCR) was utilized to measure the expression of CircRNA_0001543, IκBα, and NF-κB p65. ResultsAfter 48 hours, 30% Huangqin Qingre Chubi Capsules-containing serum significantly inhibited the proliferation of co-cultured PBMCs and FLSs, which was determined to be the optimal experimental drug-containing serum concentration. Compared with those in the normal group, the expressions of NF-κB p65 mRNA, IκBα mRNA, IL-1β, and TNF-α in the model group were significantly increased (P<0.01), while the expressions of CircRNA_0001543 mRNA, IL-10, and IL-37 were significantly decreased (P<0.01). Compared with those in the model group, the expressions of NF-κB p65 mRNA, IκBα mRNA, IL-1β, and TNF-α in the Huangqin Qingre Chubi Capsules-containing serum group were significantly decreased (P<0.05), and the expressions of CircRNA_0001543 mRNA, IL-10, and IL-37 were significantly increased (P<0.05), with the most prominent changes in the 30% drug-containing serum group (P<0.01). Compared with that in the si-NC group, the expression of CircRNA_0001543 was significantly reduced in the si-CircRNA_0001543 group (P<0.01). Compared with that in the OE-NC group, the expression of CircRNA_0001543 was significantly increased in the OE-CircRNA_0001543 group (P<0.01), indicating that the si-CircRNA_0001543 and OE-CircRNA_0001543 plasmids were successfully transfected. Based on the optimal drug-containing serum of Huangqin Qingre Chubi Capsules, si-CircRNA_0001543 transfection led to significantly increased expressions of NF-κB p65 mRNA, IκBα mRNA, IL-1β, and TNF-α and decreased the expressions of IL-10 and IL-37 (P<0.01). In contrast, OE-CircRNA_0001543 transfection significantly decreased the expressions of NF-κB p65 mRNA, IκBα mRNA, IL-1β, and TNF-α (P<0.01) and increased the expressions of IL-10 and IL-37 (P<0.01). ConclusionHuangqin Qingre Chubi capsules-containing serum can improve immune inflammation in AS by increasing the expression of CircRNA_0001543, regulating the NF-κB pathway, suppressing pro-inflammatory cytokines, and enhancing anti-inflammatory cytokine expression.
2.Study on the Predictive Value of Serum Hsa-miR-30c-5p Level Expression in Patients with Type 2 Diabetes Mellitus for Micro vascular Complications
Xiaoqin XIONG ; Xiaoyu FENG ; Aijun MAO ; Ding MA ; Yajun SHI
Journal of Modern Laboratory Medicine 2024;39(5):63-68
Objective To explore the predictive value of serum homo sapiens microRNA(hsa-miR)-30c-5p expression in patients with type 2 diabetes mellitus(T2DM)for microvascular complications.Methods A total of 205 T2DM patients admitted to Bazhong Central Hospital from May 2021 to September 2022 were selected as the diabetes group,and the diabetes group was further divided into diabetes with combined group(n=124)and non combined group(n=81)according to the microvascular complications of the patients.In addition,205 healthy people who underwent physical examination during the same period were selected as the control group.The expression of hsa-miR-30c-5p in serum was detected by reverse transcription-polymerase chain reaction(RT-PCR)and compared.The factors affecting microvascular complications were analyzed using multivariate logistic regression analysis,and receiver operating characteristic(ROC)curves were plotted to predict the value of serum hsa-miR-30c-5p expression in predicting microvascular complications in T2DM patients.Results The expression of serum hsa-miR-30c-5p in the combined group(0.58±0.06)and the non-combined group(0.72±0.08)were lower than that in the control group(0.89±0.21),and the differences were significant(t=16.038,7.079,all P=0.001).The combined group was lower than the non-combined group,and the difference was significant(t=14.289,P=0.001).The course of diabetes[(OR(95%CI):3.873(2.976~4.770)],uric acid[(OR(95%CI):2.125(1.211~3.040)]and glycosylated hemoglobin[(OR(95%CI):2.680(1.745~3.616)]were independent risk factors for microvascular complications in T2DM patients(all P<0.05),while the time within the target range of glucose[(OR(95%CI):0.491(0.135~0.846)]and serum hsa-miR-30c-5p[(OR(95%CI):0.532(2.976~4.770)]were protective factors for microvascular complications in T2DM patients(all P<0.05).The sensitivity,specificity and area under the curve(95%CI)of serum hsa-mir-30c-5p expression in predicting microvascular complications in T2DM patients were 81.45%,85.19%and 0.802(0.741~0.854),respectively.Conclusion The expression of serum hsa-miR-30c-5p in patients with T2DM is abnormally reduced,and serum hsa-miR-30c-5p is a protective factor for microvascular complications in patients with T2DM.It may have a certain predictive value for microvascular complications in patients with T2DM.
3.Research progress of the role of intestinal microbiota-mediated bile acids in inflammatory bowel disease
Xixi XIA ; Keke DING ; Huiheng ZHANG ; Xufei PENG ; Yimin SUN ; Yajun TANG ; Xiaofang TANG
Journal of Shanghai Jiaotong University(Medical Science) 2024;44(7):839-846
It is estimated that approximately seven million people worldwide are affected by inflammatory bowel disease(IBD),causing a huge burden on healthcare systems and society.In the occurrence,progression,and treatment of IBD,the intestinal microbiota and its key metabolic product,bile acids,play a crucial role.The intestinal microbiota not only participates in the biotransformation of bile acids,enriching the diversity of bile acids,but also regulates their synthesis and transport through the farnesoid X receptor(FXR).Meanwhile,bile acids contribute to regulating the structure and function of the intestinal microbiota by supporting microbial diversity,exerting direct toxicity,participating in indirect antimicrobial pathways,and influencing microbial metabolic capabilities.Furthermore,under normal physiological conditions,intestinal microbiota-derived bile acids facilitate the repair process of the intestinal epithelial barrier.They also promote the balance of the immune system by modulating the functions of various immune cells including helper T(Th)cells 17,regulatory T(Treg)cells,CD8+T cells and natural killer T(NKT)cells,thereby slowing down the development of IBD.This article focuses on exploring the role of intestinal microbiota and bile acids in the onset and progression of IBD,and investigating new effective treatment strategies by targeting intestinal microbiota and bile acids,such as bile acid receptor modulators,probiotics,prebiotics,fecal microbiota transplantation(FMT),and phage therapy.
4.The effect of B-ultrasound-guided lower abdominal transverse nerve block on patients after radical resection of colorectal cancer
Peipei KANG ; Zhiyun LI ; Shan ZHU ; Yajun DING ; Lei ZHANG
Chinese Journal of Postgraduates of Medicine 2024;47(11):983-987
Objective:To investigate the effect of B-ultrasonic-guided lower abdominal transverse nerve block on patients after radical resection of colorectal cancer.Methods:Ninety-eight patients who were underwent radical resection of colorectal cancer in Affiliated Tumor Hospital of Nantong University from September 2020 to September 2023, according the numerical random table method divided into two groups, 49 patients in control group were treated with intravenous controlled analgesia pump, 49 patients in observation group were treated with lower abdominal transverse nerve block guided by B-ultrasound, the analgesic effect, oxidative stress reaction, immune function and surgical complications were compared between the two groups.Results:The visual analogue score (VAS) 2, 12, 24, 48 and 72 h after operation in observation group was significantly lower than that in control group: (1.03 ± 0.25) scores vs. (1.32 ± 0.28) scores, (2.78 ± 0.42) scores vs. (3.52 ± 0.47) scores, (2.69 ± 0.38) scores vs. (3.21 ± 0.44) scores, (2.11 ± 0.31) scores vs. (2.65 ± 0.32) scores and (2.05 ± 0.27) scores vs. (2.43 ± 0.31) scores, and there was statistical difference ( P<0.05). The superoxide enzyme (SOD) in observation group was significantly higher than that in control group: (72.65 ± 4.28) kU/L vs. (67.58 ± 4.31) kU/L, and the malondialdehyde (MDA) was significantly lower than that in control group: (16.51 ± 1.23) mg/L vs. (18.82 ± 1.21) mg/L, and there were statistical differences ( t = 5.84 and 8.57, P<0.05). The CD 4+ and CD 8+ in observation group were significantly higher than those in control group (0.334 ± 0.03 vs. 0.282 ± 0.032 and 0.292 ± 0.030 vs. 0.252 ± 0.030), and there were statistical differences ( t = 7.90 and 6.55, P<0.05). The complication rate in observation group was significantly lower than that in control group: 4.08% (2/49) vs. 18.37% (9/49), and there was statistical difference ( P<0.05). Conclusions:B-ultrasound-guided lower transverse abdominal nerve block can further enhance the analgesic effect, effectively improve the oxidative stress reaction and immune function, as well as reduce the occurrence of surgical complications in patients with colorectal cancer after radical surgery.
5.Prednisone combined with icariin enhances the therapeutic effect of steroid resistant nephrotic syndrome
Shuwen DUAN ; Yajun WEI ; Tiankai WU ; Xiaohui WANG ; Zhaoran DING ; Can LIU ; Enlai DAI
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(10):1081-1090
AIM:To explore the therapeutic effect and possible molecular mechanisms of prednisone combined with icariin(ICA)on hormone resistant nephrotic syndrome(SRNS).METHODS:In the in vi-vo experiment,rats were divided into control group,SRNS group,prednisone group,and P+I group.Each group was given corresponding drugs for 6 weeks.Detection of 24-hour urinary protein in rats using CBB;The blood biochemistry analyzer de-tects rat albumin,total cholesterol,triglycerides,creatinine,and urea nitrogen;HE and Masson were used to detect morphological changes in rat kidney tissue;Immunohistochemical detection of GR-α,GR-β,NLRP3,caspase-1,GSDMD,IL-1β.In the in vi-tro experiment,HK-2 cell injury model with doxoru-bicin,divided into control group,SRNS group,pred-nisone group,P+l group.GR-α,GR-β,NLRP3,cas-pase-1,GSDMD were detected by Rt-PCR and West-ern blot.RESULTS:In the in vivo experiment,com-pared with the control group,the SRNS group showed weight loss,increased 24-hour urine pro-tein,decreased albumin,increased total cholester-ol,triglycerides,creatinine,and urea nitrogen,renal tubular atrophy,increased renal interstitial area,sig-nificant infiltration of inflammatory cells,fibrous tis-sue proliferation,and GR-β,NLRP3,caspase-1,GSD-MD,IL-1 β in renal tissue decreased(P<0.01);Com-pared with the SRNS group,the combined group showed weight gain,decreased 24-hour urine pro-tein,increased albumin,decreased total cholester-ol,triglycerides,creatinine,and urea nitrogen,re-duced renal tubular atrophy,reduced interstitial in-flammatory cell infiltration,reduced fibrosis,and and GR-α,NLRP3,caspase-1,GSDMD in renal tissue decreased increased(P<0.01).In vitro experiments,compared with the control group,the model group showed GR-β,NLRP3,caspase-1,and GSDMD in-creased(P<0.01),GR-α decreased(P<0.01);Com-pared with the SRNS group,GR-β,NLRP3,caspase-1,and GSDMD decreased(P<0.01),GR-α increased in the P+I group.CONCLUSION:The combination of prednisone and ICA has a protective effect on the kidneys of SRNS rats and can improve the therapeu-tic effect.The mechanism may be related to the NL-RP3/Caspase-1/GSDMD pathway.
6.Effect and mechanism of paeoniflorin on LPS-induced bronchial epithelial cell inflammatory response based on NF-κB pathway
Fang HE ; Min DING ; Haining ZHEN ; Yajun CHEN ; Xinxin XUE
Chinese Journal of Immunology 2024;40(7):1387-1391
Objective:To explore the effect of paeoniflorin on LPS-induced bronchial epithelial cell inflammatory response by regulating NF-κB pathway.Methods:BEAS-2B cells were cultured in vitro for paeoniflorin toxicity assay and concentration screening.BEAS-2B cells were divided into control group,LPS group,LPS+CAPE group,LPS+PF group and LPS+CAPE+PF group.Inflammatory responses were induced in BEAS-2B cells using LPS(1 μg/ml),and cell viability was detected by CCK-8 assay after administration of paeoniflorin or CAPE interventions.Apoptosis and cell cycle were detected by flow cytometry.The levels of IFN-γ,IL-4,IL-17C and IL-10 were detected by ELISA.The protein expression levels of p53,Bcl-2,Bax,Cyclin1,NF-κB and p-p65 were detected by Western blot.Results:Paeoniflorin increased cell viability,inhibited apoptosis,increased IFN-γ and IL-10 levels(P<0.01),de-creased IL-4 and IL-17C levels(P<0.01),down-regulated the protein expression levels of p53,Bax,NF-κB and p-p65(P<0.01),and up-regulated the protein expression levels of Bcl-2 and Cyclin1(P<0.01).The effect of paeoniflorin was more significant after the intervention of NF-κB inhibitor CAPE.Conclusion:Paeoniflorin reduces LPS-induced inflammatory factor levels in bronchial epithelial cells by regulating the NF-κB pathway,thereby suppressing the asthmatic inflammatory response.
7.Non-contact vital signs monitoring systems in the ICU based on imaging photoplethysmography technology
Chen ZHU ; Danli CAI ; Ying SHI ; Caiping SONG ; Yajun MAO ; Jianhui DING ; Qiangfang LU ; Dafen WANG ; Shuo ZHANG ; Lingcong WANG
Chinese Journal of Physical Medicine and Rehabilitation 2024;46(9):825-829
Objective:To explore the application of photoplethysmography (iPPG) for contactless vital signs monitoring in the intensive care unit (ICU).Methods:Ten tracheostomy patients in intensive care had their heart rate, oxygen saturation, and diastolic and systolic pressures monitored using iPPG technology and a 24-hour bedside monitor. The readings included periods at rest, during turning, during suctioning, and when undergoing vigorous physical therapy and occupational therapy. The monitoring lasted 3 consecutive days. The data collected by the two methods were compared to analyze the accuracy of the contactless vital signs monitoring system.Results:The oxygen saturation readings of the two systems showed no significant differences. The heart rates, diastolic pressures, and systolic pressures did, however, differ significantly.Conclusions:In the situations tested, contactless monitoring of oxygen saturation is effective, but there is still significant room for improvement in the three indicators of heart rate, systolic pressure, and diastolic pressure.
8.Effects of ACEI/ARB Agent on the Prognosis of Patients with Advanced Gastric Cancer Complicated with Hypertension
Yajun QI ; Haiying DING ; Guonong YANG ; Zeng WANG ; Peihua LUO
China Pharmacy 2017;28(32):4517-4520
OBJECTIVE:To explore the effects of angiotensin converting enzyme inhibitors (ACEI)/angiotensin Ⅱ receptor blockers (ARB) on the prognosis in patients with advanced gastric cancer complicated with hypertension.METHODS:By retrospective study,clinical information of histologically confnrned Ⅲ-Ⅳ stage advanced gastric cancer patients with hypertension receiving 2 cycles of first-line chemotherapy plan containing fluorouracil at least were collected from Zhejiang Tumor Hospital during Jan.1st,2009-Dec.31th,2012.The data were summarized and analyzed.The patients receiving ACEI/ARB drugs were included in trial group,and the patients receiving other antihypertensive drugs were included in control group.Telephone follow-up was conducted in 2 groups.RESULTS:A total of 124 patients meeting the inclusion criteria were enrolled in this study.There was no statistical significance in age,gender,smoking history,ECOG score,family history of disease,family history of tumor,TNM staging,pathological type,tumor site or chemotherapy plan between trial group (23 cases) and control group (101 cases),except for history of alcohol intake (P>0.05).Total response rate of first-line chemotherapy was 73.9% in trial group,which was significantly higher than 41.6% of control group,with statistical significance (P<0.05).There was no statistical significance in 1-year or 3-year survival rate between 2 groups (P>0.05).Median survival time of trial group was 669 d,which was significantly longer than 410 d of control group,with statistical significance (P<0.05).Results of COX regression analysis showed that therapeutic efficacy of first-line chemotherapy influenced the survival of patients in different antihypertensive agent groups (P<0.05).CONCLUSIONS:Compared with other antihypertensive drugs,ACEI/ARB agents significantly improve response rate of first-line chemotherapy in patients with advanced gastric cancer complicated with hypertension,significantly prolong median survival time,but have no significant effect on survival rate;at the same time,statistical comparison of definite drinking amount are absent.It needs to be further confirmed by multi-center,prospective and large-scale sample.
9.Preliminary clinical observation of concurrent radiochemotherapy for 31 patients with stage Ⅱ nasal cavity natural killer/T cell lymphoma
Lu HUANG ; Yunhong HUANG ; Tao WU ; Yunfei HU ; Ling DING ; Yajun MIAO ; Yongling WANG
Journal of Leukemia & Lymphoma 2015;24(8):475-478,482
Objective To observe the primary clinical effect of concurrent radiochemotherapy for patients with nasal cavity natural killer (NK)/T cell lymphoma and to analyze the prognostic factors.Methods 31 primary untreated patients with stage Ⅱ nasal cavity NK/T cell lymphoma were enrolled for this study.All patients underwent concurrent radiochemotherapy with intensity-modulated radiotherapy technique + asparaginase based chemotherapeutic agents and adjuvant chemotherapy.Results The main toxicities were mouth mucocitis,myelosuppression and xerosmia at grade 1 or 2.31 patients achieved good clinical shortterm effect with high local complete remission rate at the 3rd month after radiotherapy [83.9 % (26/31)],and the 2-year overall survival rate was 77 %.Univariate and multivariate analysis suggested IPI score and clinical short-term effect were the significant independent survival prognostic factors (P < 0.05).Conclusions Concurrent radiochemotherapy for stage Ⅱ nasal cavity NK/T cell lymphoma can be well tolerated by patients with mild toxicities,and can improve both clinical short-term effect and overall survival by high local complete remission rate.IPI score and clinical short-term effect are the important survival prognostic factors.
10.The changes of serum miR-21 expression level in patients with HCC before and after TACE and its ;clinical significance
Yilang WANG ; Yafei WANG ; Liang ZHANG ; Yajun MIAO ; Zhuo CHEN ; Chen ZHOU ; Dian YIN ; Wenbin DING
Journal of Interventional Radiology 2014;(5):406-410
Objective To investigate the changes of serum miR-21 expression level in patients with HCC before and after transcatheter arterial chemoembolization (TACE) and to discuss its clinical significance. Methods Before and after TACE the levels of serum miR-21 in 42 patients with HCC and 42 healthy subjects were determined by reverse transcriptase quantitative PCR (RT-PCR), and the levels of serum AFP were also estimated by enzyme-linked immunosorbent assay (ELISA). The results were analyzed. Results The serum miR-21 level in patients with HCC was (12.9 ± 3.5) times of that in normal subjects(t=19.430 7, P < 0.01). One month after TACE, the serum miR-21 level became (7.2 ± 1.7) times of that of normal reference value, which was remarkably lower than that obtained before the treatment (t=9.493 7, P<0.01). The serum miR-21 level was closely correlated with the tumor size, the presence of tumor thrombus and HBV infection. One month after TACE the serum miR-21 levels in patient groups showing partial response, stable disease and progressive disease were (4.0 ± 0.3), (6.0 ± 1.5) and (8.6 ± 1.5) times, respectively, of that of normal reference value, and statistically significant difference existed between each other among the three groups (F=38.168, P=0.000). ROC-AUC value of MiR-21 in diagnosing HCC was 0.910 ± 0.041, which was significantly higher than that of AFP (0.860 ± 0.037, t=6.3042, P<0.01). The specificity of miR-21 in detecting HCC was 88.1%, which was remarkably higher than that of AFP (69%, χ2= 4.5253, P = 0.033).Conclusion After TACE the serum MiR-21 level in HCC patients is significantly decreased, which is very helpful in predicting the therapeutic efficacy of TACE. Therefore, MiR- 21 can be regarded as a potential molecular marker of HCC.

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