1.Study on the effect and mechanism of Qiwei dongqingye powder against bronchial asthma based on transcriptomics
Jiacheng JIN ; Wenyan CHEN ; Xin LI ; Qing XU ; Hangyu WANG ; Ke ZHANG ; Pinghua SUN ; Jinhui WANG
China Pharmacy 2026;37(5):595-601
OBJECTIVE To investigate the therapeutic effect and mechanism of Qiwei dongqingye powder (QDP) on bronchial asthma in mice. METHODS The mice were divided into blank group (normal saline), model group (normal saline), dexamethasone group (2 mg/kg), and QDP low-, medium-, and high-dose groups (200, 400, 800 mg/kg), with 14 mice in each group. Except for the blank group, mice in all other groups were given ovalbumin via intraperitoneal injection followed by aerosol inhalation to induce a bronchial asthma model. During the modeling process, mice in each group were administered corresponding drug solutions or normal saline intragastrically/intraperitoneally. After the last medication, the number of cells in the bronchoalveolar lavage fluid (BALF) of the mice was observed and counted; the pathological changes of the bronchus and lung tissue were observed; the levels of malondialdehyde (MDA), nitric oxide (NO), total superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-Px) in the lung tissue of the mice were determined, and the level of interleukin-17 (IL-17) in the BALF and serum was determined. Transcriptomics was employed to predict and validate the mechanism of action of QDP against bronchial asthma. RESULTS Compared with the model group, the total cell count, neutrophil count, lymphocyte count, and macrophage counts in the BALF of the QDP high-dose group were all significantly reduced ( P <0.05); the levels of MDA and NO in the lung tissue, and the levels of IL-17 in the BALF and serum were all decreased significantly ( P <0.05); the levels of T-SOD and GSH-Px were significantly increased ( P <0.05); the arrangement of lung tissue cells tended to normalize, with reduced infiltration of inflammatory cells and decreased exfoliation of bronchial simple columnar epithelial cells. The transcriptomic results revealed that the differentially expressed genes were B-cell receptor signaling pathway, nuclear factor κB (NF-κB) signaling pathway, ferroptosis signaling pathway, and others. Further validation revealed that, compared with the model group, the expression levels of NF-κB p65 and chemokine ligand 20, as well as the phosphorylation level of NF-κB inhibitor protein α, were significantly decreased in the lung tissues of the mice in all QDP groups ( P <0.05). Conversely, the protein expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1 (HO-1) were significantly increased ( P <0.05). CONCLUSIONS QDP can effectively alleviate bronchial asthma by inhibiting the NF-κB signaling pathway, activating the Nrf2/HO-1 signaling pathway, regulating oxidative stress, and reducing inflammatory responses.
2.In Vitro Anti-psoriatic Effect of Kangfuxin Liquid via Inhibiting Cell Proliferation and Migration Ability and Blocking JAK3/STAT3 Signaling Pathway
Shuai LI ; Xuan LIU ; Wenyan TANG ; Zhenqi WU ; Chunhui CHEN ; Dadan QIU ; Yi XU ; Chenggui ZHANG ; Jianquan ZHU ; Jiali ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):123-133
ObjectiveThis paper aims to explore the in vitro anti-psoriasis activity and potential mechanism of Kangfuxin liquid (KFX liquid), providing experimental evidence for the anti-psoriasis effect of KFX liquid. MethodsFirstly, the uninduced human immortalized keratinocyte cells (HaCaT cells) were divided into seven groups, namely the control group and KFX liquid groups with different doses (5, 10, 20, 40, 80, 160 g·L-1). After being treated with different concentrations of KFX liquid, the effect of KFX liquid on the normal cell proliferation was detected by using the cell counting kit-8 (CCK-8) method. Secondly, the uninduced HaCaT cells were divided into six groups, namely the control group and recombinant human interleukin-7A (rh-IL-7A) groups with different doses (5, 10, 50, 100, 120 g·L-1). After being treated with different concentrations of recombinant human interleukin-17A (rh IL-17A) liquid, the effect of rh IL-17A on cell proliferation was detected. The optimal induction concentration was screened. Then, normal HaCaT cells were divided into a control group and KFX liquid groups with different doses (5, 10, 20, 40, 80, 160 g·L-1). Except for the control group, the other groups established psoriasis cell models with the optimal induction concentration of rh IL-17A. After being treated with different concentrations of KFX liquid, the effects of KFX liquid on the psoriasis-like HaCaT cell proliferation were investigated. Finally, the uninduced HaCaT cells were divided into six groups, namely the control group, rh IL-17A group, methotrexate (MTX) group, and KFX liquid groups with different doses (20, 40, 80 g·L-1). Except for the control group, the other groups used the optimal induction concentration of rh IL-17A to establish psoriasis cell models. After being treated with different drugs, the cell migration levels were detected through scratch assays, and real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the relative mRNA expression levels of Ki-67 antigen (Ki67), S100 calcium-binding protein A7 (S100A7), S100 calcium-binding protein A8 (S100A8), and S100 calcium-binding protein A9 (S100A9), thereby comprehensively evaluating the in vitro anti-psoriasis activity of KFX liquid. By detecting the relative mRNA expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and chemokine-20 (CXCL-20) inflammatory-related factors in psoriasis-like HaCaT cells and the protein expression levels of Janus kinase 3 (JAK3), phosphorylated Janus kinase 3 (p-JAK3), signal transducer and activator of transcription 3 (STAT3), and phosphorylated signal transducer and activator of transcription 3 (p-STAT3), the mechanism was explored. ResultsCompared with that of control group, when treated with 80 g·L-1 KFX liquid for 72 h (P<0.05) and at different times with 160 g·L-1 KFX liquid, the HaCaT cell proliferation activity was significantly affected (P<0.01), while the other concentrations of KFX liquid had no significant differences in cell morphology and cell proliferation activity at different times, indicating that the KFX liquid is relatively safe for HaCaT cells and has no obvious toxic side effects. Compared with that of control group, when treated with different concentrations of rh IL-17A for 24 h, the HaCaT cell proliferation activity was significantly enhanced, and the cell activity was the strongest when the concentration was 100 μg·L-1 (P<0.05), with a density close to 100% and intact cell morphology, indicating that 100 μg·L-1 is the optimal concentration for inducing HaCaT cell proliferation. The results of the KFX liquid treatment on rh IL-17A-induced psoriasis-like cells show that the KFX liquid not only effectively inhibits the rh IL-17A-induced psoriasis-like HaCaT cell proliferation activity (P<0.01), but also significantly reduces the migration ability of rh IL-17A-induced psoriasis-like HaCaT cells (P<0.01), and the relative mRNA expression levels of Ki67, S100A7, S100A8, and S100A9 (P<0.01). Moreover, the KFX liquid can significantly reduce the relative mRNA expression levels of IL-1β, IL-6, and CXCL-20 in rh IL-17A-induced psoriasis-like cells (P<0.01), and significantly inhibit the phosphorylation levels of JAK3 and STAT3 proteins (P<0.05, P<0.01). ConclusionThe KFX liquid has no obvious toxicity to uninduced HaCaT cells. It can inhibit rh IL-17A-induced psoriasis-like HaCaT cell proliferation, reduce the cell migration ability, and has good in vitro anti-psoriasis activity. Its action mechanism may be related to downregulating the expression levels of inflammation-related cytokines in the JAK3/STAT3 signaling pathway and inhibiting the phosphorylation levels of JAK3 and STAT3 proteins.
3.In Vitro Anti-psoriatic Effect of Kangfuxin Liquid via Inhibiting Cell Proliferation and Migration Ability and Blocking JAK3/STAT3 Signaling Pathway
Shuai LI ; Xuan LIU ; Wenyan TANG ; Zhenqi WU ; Chunhui CHEN ; Dadan QIU ; Yi XU ; Chenggui ZHANG ; Jianquan ZHU ; Jiali ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):123-133
ObjectiveThis paper aims to explore the in vitro anti-psoriasis activity and potential mechanism of Kangfuxin liquid (KFX liquid), providing experimental evidence for the anti-psoriasis effect of KFX liquid. MethodsFirstly, the uninduced human immortalized keratinocyte cells (HaCaT cells) were divided into seven groups, namely the control group and KFX liquid groups with different doses (5, 10, 20, 40, 80, 160 g·L-1). After being treated with different concentrations of KFX liquid, the effect of KFX liquid on the normal cell proliferation was detected by using the cell counting kit-8 (CCK-8) method. Secondly, the uninduced HaCaT cells were divided into six groups, namely the control group and recombinant human interleukin-7A (rh-IL-7A) groups with different doses (5, 10, 50, 100, 120 g·L-1). After being treated with different concentrations of recombinant human interleukin-17A (rh IL-17A) liquid, the effect of rh IL-17A on cell proliferation was detected. The optimal induction concentration was screened. Then, normal HaCaT cells were divided into a control group and KFX liquid groups with different doses (5, 10, 20, 40, 80, 160 g·L-1). Except for the control group, the other groups established psoriasis cell models with the optimal induction concentration of rh IL-17A. After being treated with different concentrations of KFX liquid, the effects of KFX liquid on the psoriasis-like HaCaT cell proliferation were investigated. Finally, the uninduced HaCaT cells were divided into six groups, namely the control group, rh IL-17A group, methotrexate (MTX) group, and KFX liquid groups with different doses (20, 40, 80 g·L-1). Except for the control group, the other groups used the optimal induction concentration of rh IL-17A to establish psoriasis cell models. After being treated with different drugs, the cell migration levels were detected through scratch assays, and real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the relative mRNA expression levels of Ki-67 antigen (Ki67), S100 calcium-binding protein A7 (S100A7), S100 calcium-binding protein A8 (S100A8), and S100 calcium-binding protein A9 (S100A9), thereby comprehensively evaluating the in vitro anti-psoriasis activity of KFX liquid. By detecting the relative mRNA expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and chemokine-20 (CXCL-20) inflammatory-related factors in psoriasis-like HaCaT cells and the protein expression levels of Janus kinase 3 (JAK3), phosphorylated Janus kinase 3 (p-JAK3), signal transducer and activator of transcription 3 (STAT3), and phosphorylated signal transducer and activator of transcription 3 (p-STAT3), the mechanism was explored. ResultsCompared with that of control group, when treated with 80 g·L-1 KFX liquid for 72 h (P<0.05) and at different times with 160 g·L-1 KFX liquid, the HaCaT cell proliferation activity was significantly affected (P<0.01), while the other concentrations of KFX liquid had no significant differences in cell morphology and cell proliferation activity at different times, indicating that the KFX liquid is relatively safe for HaCaT cells and has no obvious toxic side effects. Compared with that of control group, when treated with different concentrations of rh IL-17A for 24 h, the HaCaT cell proliferation activity was significantly enhanced, and the cell activity was the strongest when the concentration was 100 μg·L-1 (P<0.05), with a density close to 100% and intact cell morphology, indicating that 100 μg·L-1 is the optimal concentration for inducing HaCaT cell proliferation. The results of the KFX liquid treatment on rh IL-17A-induced psoriasis-like cells show that the KFX liquid not only effectively inhibits the rh IL-17A-induced psoriasis-like HaCaT cell proliferation activity (P<0.01), but also significantly reduces the migration ability of rh IL-17A-induced psoriasis-like HaCaT cells (P<0.01), and the relative mRNA expression levels of Ki67, S100A7, S100A8, and S100A9 (P<0.01). Moreover, the KFX liquid can significantly reduce the relative mRNA expression levels of IL-1β, IL-6, and CXCL-20 in rh IL-17A-induced psoriasis-like cells (P<0.01), and significantly inhibit the phosphorylation levels of JAK3 and STAT3 proteins (P<0.05, P<0.01). ConclusionThe KFX liquid has no obvious toxicity to uninduced HaCaT cells. It can inhibit rh IL-17A-induced psoriasis-like HaCaT cell proliferation, reduce the cell migration ability, and has good in vitro anti-psoriasis activity. Its action mechanism may be related to downregulating the expression levels of inflammation-related cytokines in the JAK3/STAT3 signaling pathway and inhibiting the phosphorylation levels of JAK3 and STAT3 proteins.
4.Investigating the effects and mechanisms of Yiqi Jiedu Decoction in protecting against ionizing radiation—induced small intestinal functional damage in mice based on ferroptosis
Yan WANG ; Minhao XU ; Wenyan ZHANG ; Yuankai GAO ; Qing XU ; An WANG ; Wenhui XU ; Sumin HU
Space Medicine & Medical Engineering 2025;36(5):389-395
Objective To observe the protective effects of Yiqi Jiedu Decoction on ionizing radiation-induced small intestinal functional injury in mice,and explore whether it alleviates such injury by inhibiting small intestinal ferroptosis,thereby providing scientific support for the discovery and development of intestinal radiation protection drugs in aerospace medicine.Methods A total of 378 male Balb/c mice were randomly divided into 7 groups:blank control group,model group,positive drug group,high-dose Yiqi Jiedu Decoction group,low-dose Yiqi Jiedu Decoction group,Liproxstatin-1 pre-irradiation administration group,and Liproxstatin-1 post-irradiation administration group,with 54 mice in each group.Each group was further divided into 3 batches,with 18 mice per batch.Seven days after preventive administration,all groups except the blank control group were subjected to a single whole-body irradiation with 2.0 Gy 60Co γ-rays.The general condition and morphological structure of the small intestine were observed at 1,3,and 7 days post-irradiation.The small intestinal charcoal propulsion rate,serum D-xylose content,and lactic acid content were measured,along with the levels of Fe,LPO,MDA,GSH,and SOD activity in the small intestine.Results Yiqi Jiedu Decoction could mitigate the decrease in body weight of mice after 2.0 Gy 60Co γ-ray irradiation,improve the morphological structure of the small intestine,reduce the small intestine charcoal propulsion rate,increase serum D-xylose levels,and decrease total serum lactate levels.It also alleviated mitochondrial shrinkage in the small intestine and reduced the contents of Fe and MDA in small intestine tissues.Conclusion Yiqi Jiedu Decoction may alleviate ionizing radiation-induced small intestinal functional injury by inhibiting ferroptosis in the small intestine,providing a new strategy for intestinal radiation injury in deep space exploration missions such as manned spaceflight.
5.Correlation between psoas major index of diffuse large B-cell lymphoma patients and prognosis after R-CHOP chemotherapy
Wei JIANG ; Ke ZHAO ; Cuiping BAO ; Yongchun JIA ; Zugui LI ; Wenyan ZHANG ; Liang XU
Chinese Journal of Medical Imaging Technology 2025;41(7):1125-1128
Objective To explore the correlation between psoas major index(PMI)in patients with diffuse large B-cell lymphoma(DLBCL)and prognosis after rituximab,cyclophosphamide,doxorubicin,vincristine,prednisone(R-CHOP)chemotherapy.Methods Totally 148 DLBCL patients who received R-CHOP chemotherapy were retrospectively enrolled.Based on axis CT image at L3 vertebral transverse process,bilateral psoas muscles were delineated,and PMI was calculated.Then the patients were divided into muscle reduction group(≤lower quartile,n=37)and control group(>lower quartile,n=111)according to the lower quartile of PMI.Clinical and imaging data were compared between groups,and the correlations of PMI with the above indexes were analyzed.The impact factors of prognosis of DLBCL were observed.Results Significant differences of body mass index(BMI),Eastern Cooperative Oncology Group(ECOG)score,international prognostic index(IPI),the proportion of B symptoms,mortality rate,3-year survival rate,β2-microglobulin,hemoglobin,albumin and PMI were found between groups(all P<0.05).PMI of DLBCL patient was negatively correlated with age,ECOG score,IPI,β2-microglobulin and lactate dehydrogenase(LDH)(r=—0.126,-0.225,—0.262,—0.232,—0.229,all P<0.05),but positively correlated with BMI and hemoglobin(r=0.282,0.376,both P<0.05).The age,ECOG score,IPI and PMI were all independent impact factors of overall survival in DLBCL patients(all P<0.05).Conclusion PMI was an independent impact factor of prognosis of DLBCL after R-CHOP chemotherapy.
6.Effect of dexmedetomidine on viability of dopaminergic neurons in ventral tegmental area of morphine-addicted mice
Guanhua LU ; Jian HE ; Wenyan PAN ; Fuyu WANG ; Hongsen XU ; Huailong ZHANG ; Wanyou HE ; Hanbing WANG
Chinese Journal of Anesthesiology 2025;45(7):834-839
Objective:To evaluate the effect of dexmedetomidine on the viability of dopaminergic neurons in the ventral tegmental area (VTA) of morphine-addicted mice.Methods:Experiment Ⅰ Thirty SPF healthy adult male C57BL/6 mice, aged 8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) using the random number table method: normal saline group (NS group), dexmedetomidine 50 μg/kg group (DEX50 group), and dexmedetomidine 100 μg/kg group (DEX100 group). A morphine addiction model was established by intraperitoneal injection of increasing doses of morphine (10, 20, 30, 40, 50 and 50 mg/kg) for 6 consecutive days in mice. After the successful establishment of the model, dexmedetomidine 50 and 100 μg/kg were intraperitoneally injected for 14 consecutive days in group DEX50 and group DEX100 respectively, while normal saline was given instead in group C. The conditioned place preference (CPP) experiment was conducted every other day. Experiment Ⅱ Thirty SPF healthy adult male C57BL/6 mice, aged 8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) by the random number table method: control group (C group), morphine group (Mor group) and dexmedetomidine 50 μg/kg group (DEX50 group). Normal saline was intraperitoneally injected for 10 consecutive days in group C. Morphine with increasing doses was intraperitoneally injected for 6 days, and then normal saline was intraperitoneally injected for 4 consecutive days in group Mor. Morphine with increasing doses was intraperitoneally injected for 6 days, and then dexmedetomidine 50 μg/kg was intraperitoneally injected for 4 consecutive days in group DEX50. The mice were anesthetized at 90 min after the last intraperitoneal injection, brain tissues were harvested, and the corresponding brain slices of the VTA were selected for c-Fos immunofluorescence staining. Experiment Ⅲ Ten dopamine transporter-Cre recombinase mice were divided into 2 groups ( n=5 each) by the random number table method: morphine group (Mor group) and morphine+ dexmedetomidine 50 μg/kg group (Mor+ DEX group). Stereotaxic viral injection was performed in the brain. rAAV-EF1α-DIO-GCaMP6s was injected into the VTA and an optical fiber was implanted. Three weeks later, a morphine addiction model was established based on Experiment Ⅰ for the CPP experiment, morphine was intraperitoneally injected in group Mor, and morphine and dexmedetomidine were intraperitoneally injected in group Mor+ DEX. The viral fluorescence signals were recorded at 5 min before and 20 min after the drug administration in the three groups. Results:Experiment Ⅰ There was no statistically significant difference in the CPP scores after developing the morphine addiction model among the three groups ( P>0.05). Compared with group NS, the CPP scores were significantly decreased at 4-14 days of the continuous administration in group DEX50 and group DEX100 ( P<0.05). Experiment Ⅱ Compared with group C, the number of c-Fos positive cells in the VTA was significantly increased in group Mor ( P<0.05). Compared with group Mor, the number of c-Fos positive cells in the VTA was significantly decreased in group DEX ( P<0.05). Experiment Ⅲ Compared with that before administration, the calcium signals of dopaminergic neurons in the VTA were significantly enhanced in group Mor ( P<0.05), and no statistically significant difference was found in the calcium signals of dopaminergic neurons in the VTA in group Mor+ DEX ( P>0.05). Compared with group Mor, no statistically significant difference was found in the calcium signals of dopaminergic neurons in the VTA before drug administration ( P>0.05), and the calcium signals of dopaminergic neurons in the VTA were significantly weakened after administration in group Mor+ DEX ( P<0.05). Conclusions:The mechanism by which dexmedetomidine promotes the extinction of morphine addiction is related to the inhibition of the viability of dopaminergic neurons in the VTA of mice.
7.Develop an evaluation system for quality palliative care based on the guidelines of National Consensus Project for Quality Palliative Care
Minling XU ; Weiying DENG ; Lijun ZHAN ; Wenyan CHEN ; Lan WAN
Modern Clinical Nursing 2025;24(9):57-65
Objective To develop an evaluation system for quality palliative care and provide an evaluation tool for quality palliative care service.Methods A preliminary evaluation system was drafted by using literature review and group discussion,aligning with the U.S.National Consensus Project for Quality Palliative Care(NCP).The system was revised and refined by two rounds of Delphi consultation with 15 palliative care experts(including specialists in clinical practice,nursing management,nursing research and education)from Tier-IIIA hospitals in Guangdong Province.Indicator weights were determined via consensus.Results Both rounds of expert consultation achieved 100.00%response rates.Expert authority coefficient(Cr)was 0.855.The importance scores of the level-1,level-2 and level-3 indicators of the second round of expert consultation were 4.90-5.00,4.80-5.00 and 4.37-5.00,respectively.Coefficients of variation were 0-0.06,0-0.1 and 0-0.19,respectively.The full score ratio ranged between 0.93 and 1.00,0.8 and 1.00,and 0.67 and 1.00.Kendall's W coefficients were 0.214,0.287 and 0.245,respectively(all P<0.01).The nine level-1 indicators were identified as care structure and process,physiological care,psychological care,social care,mental care,cultural care,end-of-life care,ethical care and quality improvement,with the weight coefficients of 0.123,0.153,0.110,0.106,0.098,0.082,0.119,0.092 and 0.117,respectively.The final evaluation system for quality palliative care included 9 indicators in level-1,22 in level-2 and 69 in level-3.Conclusion The evaluation system for quality palliative care developed on the basis of NCP is scientifically innovative and valid in content.Further studies are required to evaluate its validity..
8.Association between albumin and recompensation in patients with hepatitis B/C virus-related decompensated liver cirrhosis
Danqing XU ; Yingyuan ZHANG ; Jingru SHANG ; Caifen SA ; Wenyan LI ; Li LIU ; Zhijian DONG
Journal of Clinical Hepatology 2025;41(11):2323-2328
ObjectiveTo investigate the association between albumin (Alb) and recompensation by comparing recompensation rate between hepatitis B/C virus-related decompensated liver cirrhosis patients with different Alb levels, and to provide guidance for the identification and management of high-risk patients in clinical practice. MethodsRelated clinical data were collected from 734 patients with hepatitis B/C virus-related decompensated liver cirrhosis who attended The Third People’s Hospital of Kunming from January 1, 2016 to December 31, 2022, and they were divided into three groups based on the level of Alb. The linear regression analysis and chi-square test were used for trend tests. The Kaplan-Meier curve was plotted for the cumulative incidence rate of recompensation in the three groups, and the log-rank test was used for comparison between groups. A Cox proportional-hazards regression model analysis was used to investigate the association between Alb and recompensation in patients with hepatitis B/C virus-related decompensated liver cirrhosis. ResultsAmong the 734 patients with hepatitis B/C virus-related decompensated liver cirrhosis, 270 achieved recompensation, with a recompensation rate of 36.8%. All patients had a median Alb level of 29.90 (25.90 — 34.80) g/L on admission, and according to the level of Alb, they were divided into <25.9 g/L group with 177 patients, 25.9 — 34.8 g/L group with 377 patients, and >34.8 g/L group with 180 patients; 36 patients (20.3%) in the <25.9 g/L group, 138 (36.6%) in the 25.9 — 34.8 g/L group, and 96 (53.3%) in the >34.8 g/L group achieved recompensation, and the recompensation rate increased with the increase in Alb level (χ2=41.730, P<0.001). After adjustment for all confounding factors, compared with the <25.9 g/L group, there was a significant increase in the incidence rate of recompensation in the 25.9 — 34.8 g/L group (hazard ratio [HR]=1.842, 95% confidence interval [CI]: 1.274 — 2.663) and the >34.8 g/L group (HR=2.336, 95% CI: 1.575 — 3.463). The Kaplan-Meier survival analysis showed that there was a significant difference in the cumulative incidence rate of recompensation between the three groups (χ2=41.632, P<0.001). ConclusionAlb level is an influencing factor for recompensation in patients with hepatitis B/C virus-related decompensated liver cirrhosis, and the recompensation rate increases with the increase in Alb level.
9.Effects of lncRNA DUXAP8 in lung cancer A549 cells-derived exosomes on lung cancer cell growth and its mechnism
Xiaoshuang HE ; Lina XU ; Mei CUI ; Yu ZHAO ; Bei WANG ; Zheng HUANG ; Yuchao WANG ; Wenyan XIN ; Chao WU
Journal of Jilin University(Medicine Edition) 2025;51(4):958-967
Objective:To discuss the effect of long non-coding RNA(lncRNA)DUXAP8 in exosomes(Exo)derived from the lung cancer A549 cells on the growth and immune escape of the lung cancer cells,and to clarify the mechanism.Methods:The human lung cancer cell line A549 was cultured,and its exosomes were extracted and identified.The A549 cells were treated with PKH67-labeled Exo to observe the uptake of Exo by A549 cells.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression level of lncRNA DUXAP8 in A549 cells before and after Exo treatment.The A549 cells were divided into control group(no treatment),Exo group(A549 cells treated with Exo),Exo+sh-NC group(A549 cells treated with Exo and then transfected with sh-NC),and Exo+sh-DUXAP8 group(A549 cells treated with Exo and then transfected with sh-DUXAP8).RT-qPCR method was used to detect the expression level of lncRNA DUXAP8 in A549 cells in various groups;colony formation assay was used to detect the colony formation abilities of the A549 cells in various groups;5-ethynyl-2'-deoxyuridine(EdU)staining method was used to detect the proliferation abilities of the A549 cells in various groups.After co-culturing A549 cells in various groups with human peripheral blood lymphocytes,flow cytometry was used to detect the percentages of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in various groups;3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)method was used to detect the killing rates of human peripheral blood lymphocytes on the A549 cells in various groups.Results:The diameter of Exo vesicles was 50-150 nm,and the exosome-specific marker proteins cluster of differentiation 63(CD63),cluster of differentiation 9(CD9),tumor susceptibility gene 101(TSG101),and heat shock protein 70(HSP70)were positively expressed,indicating successful exosome extraction.A549 cells efficiently took up PKH67-labeled Exo.The RT-PCR results showed that compared with A549 cells cultured alone,the expression level of lncRNA DUXAP8 in the A549 cells was increased after treatment with Exo derived from A549 cells(P<0.05).compared with control group,the expression level of lncRNA DUXAP8 in the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the expression level of lncRNA DUXAP8 in the A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there were no significant difference in the expression level of IncRNA DUXAP8 in the cells in Exo+sh-NC group(P>0.05).The colony formation assay results showed that compared with control group,the number of colony formation of the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the number of colony formation of the A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there was no significant difference in the number of colony formation of the A549 cells in Exo+sh-NC group(P>0.05).The EdU staining results showed that compared with control group,the EdU-positive rate of the A549 cells in Exo group was increased(P<0.05);compared with Exo group,the EdU-positive rate in A549 cells in Exo+sh-DUXAP8 group was decreased(P<0.05),while there was no significant difference in the EDU-positive rate in the cells in Exo+sh-NC group(P>0.05).The flow cytometry results showed that compared with control group,the percentage of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in Exo group was decreased(P<0.05);compared with Exo group,the percentage of activated CD8+T lymphocytes in the human peripheral blood lymphocytes in Exo+sh-DUXAP8 group was increased(P<0.05),while there was no significant difference in the percentage of activated CD8+T lymphaytes in Exo+sh-NC group(P>0.05).The MTT assay results showed that compared with control group,the killing rate of human peripheral blood lymphocytes on the A549 cells in Exo group was decreased(P<0.05);compared with Exo group,the killing rate of human peripheral blood lymphocytes on A549 cells in Exo+sh-DUXAP8 group was increased(P<0.05),while no significant difference was observed in Exo+sh-NC group(P>0.05).Conclusion:The lncRNA DUXAP8 in exosomes derived from the lung cancer A549 cells promotes the proliferation of lung cancer cells and tumor immune escape.
10.Prospective randomised controlled observation of tympanic chamber injection of gangliosides in the treatment of refractory sudden deafness.
Mengyuan WANG ; Qi DONG ; Yuqin XU ; Yaxiu LI ; Jing LIU ; Jie LI ; Wenyan ZHU ; Wandong SHE
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2025;39(3):218-222
Objective:This study aimed to evaluate the therapeutic effect of intratympanic injection of ganglioside in patients with refractory sudden deafness. Methods:A total of 120 patients with sudden deafness, aged 18-65 years, whose onset was within 11-42 days, failed to respond to conventional treatment, and had an average hearing threshold(500-4 000 Hz)>60 dB were selected. They were prospectively and randomly divided into a control group of 61 cases and an experimental group of 59 cases. The control group was treated according to the recommended protocol of the Chinese Medical Association(postauricular injection of methylprednisolone), while the experimental group was treated with intratympanic injection of monosialotetrahexosylganglioside sodium+postauricular injection of methylprednisolone. Both groups were simultaneously administered oral ginkgo biloba extract and citicoline tablets. Hearing was re-examined two weeks after the completion of treatment, and the therapeutic effects of the two different treatment methods were compared and analyzed. Results:The effective rate was 29.51% in the control group and 54.24% in the experimental group(P<0.01). The average hearing threshold improved by 11.57 dB HL in the control group and 22.50 dB HL in the experimental group(P<0.05). Conclusion:The combination of postauricular injection of methylprednisolone and intratympanic injection of ganglioside is more effective than postauricular injection of methylprednisolone alone in the treatment of refractory sudden deafness. The earlier the treatment, the better the therapeutic effect.
Humans
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Middle Aged
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Hearing Loss, Sudden/drug therapy*
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Adult
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Prospective Studies
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Young Adult
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Aged
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Adolescent
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Male
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Female
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Injection, Intratympanic
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Gangliosides/administration & dosage*
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Methylprednisolone/therapeutic use*
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Treatment Outcome

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