1.Mechanism of Shenqi Dihuangtang in Blocking Renal Fibrosis Injury in Diabetic Kidney Disease Mediated by Epithelial-mesenchymal Transition Through Inhibiting TGF-β1/Smad Signaling Axis
Liangjing LIU ; Haolan LIU ; Xiaoling MAO ; Min YU ; Weitong YAN ; Chao LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):32-45
ObjectiveThis paper aims to study the potential active compound components and action mechanism of Shenqi Dihuangtang in the treatment of diabetic kidney disease (DKD) through network pharmacology and in vivo experimental verification. MethodsUltra-high-performance liquid chromatography-Q-exactive orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap MS) technology was used to clarify the main active chemical components of Shenqi Dihuangtang, and it was combined with network pharmacology methods such as gene ontology (GO) functional annotations and Kyoto encyclopedia of genes and genome (KEGG) to predict the potential action mechanism of Shenqi Dihuangtang in treating DKD. Subsequently, the DKD model of db/db male mice was established, and the mice were randomly divided into model group, low-dose Shenqi Dihuangtang group (6.10 g·kg-1), medium-dose Shenqi Dihuangtang group (12.19 g·kg-1), high-dose Shenqi Dihuangtang group (24.38 g·kg-1), and daplizin group (1.25 mg·kg-1). During the same period, C57BL/6J male mice were selected into normal group and received drug intervention for 8 weeks, respectively. During this period, the body weight and fasting blood glucose (FBG) of the mice were dynamically monitored, and oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were performed at the end of dosing. The levels of serum creatinine (SCr), blood urea nitrogen (BUN), uric acid (UA), albumin (ALB), and total protein (TP) were measured by an automatic biochemical analyzer, and 24-hour urine protein was measured by a urine protein quantitative kit. Hematoxylin-eosin (HE), periodic-acid Schiff (PAS), and Masson staining were employed to observe the renal histopathology. The expression of nephrotic protein Nephrin was observed by immunohistochemistry. Western blot was used to detect the expression of epithelial-mesenchymal transition (EMT)-related proteins such as TGF-β1, Smad2/3, α-smooth muscle actin (α-SMA), neural-cadherin (N-Cadherin), and snail protein. ResultsUPLC-Q-Exactive Orbitrap MS identified 384 active compounds in the aqueous extract of Shenqi Dihuangtang. According to oral bioavailability≥30% and the five drug-like principles, 44 key active ingredients were screened out, and 169 intersection targets highly correlated with DKD were matched. Among them, there was a significant interaction relationship between tumor necrosis factor(TNF), interleukin(IL)-6, protein kinase B(Akt)1, Caspase-3, Jun proto-oncogene (JUN), hypoxia inducible factor-1α(HIF-1α), B cell lymphoma-2(Bcl-2), matrix metallopeptidase-9(MMP-9), IL-1β, and TGF-β1. GO functional annotations were significantly enriched in cellular components such as membrane rafts, membrane microdomains, and collagen-containing extracellular matrix, molecular functions such as DNA-binding transcription factor binding, R-Smad binding, and Smad protein binding, as well as biological processes such as reactions to lipopolysaccharides(LPS), reactions to bacteria-derived molecules, and wound healing. The KEGG pathway was significantly enriched in lipids and atherosclerosis, TGF-β signaling pathway, phosphatidylinositol 3 kinase (PI3K)/Akt signaling pathway, etc. In vivo experimental results showed that the high-dose Shenqi Dihuangtang group could significantly reduce FBG levels in db/db mice (P<0.01), improve OGTT (P<0.01) and ITT (P<0.01) levels, reduce SCr (P<0.01), BUN (P<0.01), UA (P<0.01) and 24-hour BUN (P<0.01), and increase ALB (P<0.01) and TP (P<0.01) levels. Pathological staining confirmed that the high-dose Shenqi Dihuangtang group could significantly reduce the glomerular mesangial matrix area and collagen deposition (P<0.01) and upregulate the positive expression rate of Nephrin (P<0.01). Western blot results showed that the high-dose Shenqi Dihuangtang group significantly downregulated the expression of TGF-β1 (P<0.01) and Smad2/3 (P<0.01) signal molecules and inhibited the protein levels of α-SMA (P<0.01), N-Cadherin (P<0.01), and Snail (P<0.01). ConclusionShenqi Dihuangtang can inhibit the TGF-β1/Smad signaling axis and block the renal EMT process, thereby improving DKD renal fibrosis damage. Further analysis of its key active components and clinical transformation pathways is needed in the future.
2.Epidemiological characteristics and spatial-temporal aggregation of scarlet fever in Nantong City in 2009 - 2023
Chao BAO ; Junfeng MIAO ; Enhui ZHAO ; Zhenzhen LIU ; Wuhong ZHANG ; Ye WEI
Journal of Public Health and Preventive Medicine 2026;37(2):40-44
Objective To analyze the epidemiological characteristics and spatial-temporal clustering trend of scarlet fever in Nantong from 2009 to 2023, and to provide a scientific basis for scarlet fever prevention and control. Methods The incidence data of scarlet fever in Nantong from 2009 to 2023 were analyzed. Descriptive analysis, seasonal index method and Joinpoint 5.2.0 software were used to analyze epidemiological characteristics. Spatial-temporal clustering was assessed with SaTScan 10.2.5 software. Results The average annual incidence of scarlet fever in Nantong from 2009 to 2023 was 6.54/100 000. The overall morbidity rate of scarlet fever in Nantong had an increasing trend from 2009 to 2019 with an average annual percentage change of 14.55% (t=3.36,P<0.05). The cases mainly occurred during late spring to early summer and late autumn to early winter. Students, preschool children and scattered children were the main scarlet fever population. The average annual incidence of males was significantly higher than that of females (χ2=7.00, P<0.05). Rugao City, Chongchuan District and Tongzhou District were identified as high-incidence areas, accounting for 76.51% of all reported cases. Spatial-temporal scan analysis indicated that Rugao City and Chongchuan District were primary cluster areas, spanning from 2015 to 2021 (RR=3.77, LLR=1 308.07, P<0.05). Conclusion The number of reported cases of scarlet fever in Nantong City from 2009 to 2023 shows epidemic and spatial clustering, mainly concentrated in the central urban area and adjacent counties (cities). It is necessary to strengthen health education and disease surveillance in high-incidence areas, as well as in key institutions and key populations before epidemic peaks.
3.Evaluation of myopia prevention and control effect of defocus incorporated multiple segments lenses in children with monocular myopia
Lu JIANG ; Mengling PENG ; Miao MA ; Xue YUAN ; Chao ZHOU
International Eye Science 2025;25(10):1683-1687
AIM:To compare the changes in diopters and axial length after 1 a of wearing defocus incorporated multiple segments(DIMS)lenses or single vision(SV)spectacle lenses in children with monocular myopia.METHODS:In this retrospective case group study, monocular myopia children aged from 6 to 14 years old in Hankou Aier Eye Hospital from October 2020 to October 2022, who were fitted with DIMS lens(n=52)or single-vision(SV)spectacle lenses(n=49)were collected. The spherical degree of myopia eyes ranged from -4.00 D to -0.50 D and the nonmyopic eyes ranged from 0 to +1.00 D, astigmatism in all eyes ranged from 0 to -2.00 D. The DIMS lens group was classified into DIMS-myopia group(the myopic eyes)and DIMS-nonmyopia group(the nonmyopic eyes). The SV lens group was also divided into SV-myopia group and SV-nonmyopia group. The changes in spherical equivalent refraction(SER)and axial length(AL)of each group were compare before and after wearing lenses for 1 a, and variations in SER and AL of both eye among groups were analzed.RESULTS: After wearing lenses for 1 a, the changes of SER in the DIMS-myopic group and the DIMS-nonmyopic group were -0.41±0.44 and -0.26±0.54 D, respectively, and the changes of AL were 0.18±0.20 and 0.15±0.15 mm, respectively. SER changes were -0.74±0.63 and -0.70±0.68 D in SV-myopic group and SV-nonmyopic group, and AL changes were 0.30±0.28 and 0.31±0.28 mm. The changes of SER and AL in the DMS-myopic and non-myopic groups were slower than those in SV group(all P<0.05). Compared with SV lenses, wearing DIMS lenses delayed and 44.6% in myopia eyes, and 62.9% in non-myopia eyes, AL delayed by 40.0% in myopia eyes and 51.6% in non-myopia eyes. The percentage of 1-year AL change ≤0.2 mm in the DIMS-myopic group and non-myopic group was 53.9% and 65.4%, respectively, which was higher than that in the SV myopic group(34.7% and 42.9%, all P<0.05). The percentage of AL change >0.4 mm in the DIMS-myopic group and nonmyopic group was 17.3% and 7.7%, respectively, which was lower than that in the SV myopic group(32.7% and 28.6%, all P<0.05). There was no significant correlation between the change of AL and age and baseline AL in the DIMS-myopic and non-myopic groups after wearing lens for 1 a(all P>0.05); the change of AL in SV-myopic group and non-myopic group was negatively correlated with age(r=-0.446, P=0.001; r=-0.312, P=0.029), and there was no significant correlation with baseline AL(all P>0.05).CONCLUSION: DIMS lens has a good effect on myopia control and prevention in both myopia and non-myopia children with monocular myopia. Children with early pre-myopia can wear DIMS to prevent myopia.
4.Overweight and obesity among primary and middle school students in Yuyao City
ZHANG Yi ; LI Mengdi ; SHI Yafeng ; MIAO Chao
Journal of Preventive Medicine 2025;37(9):955-958
Objective:
To investigate the status of overweight and obesity among primary and middle school students in Yuyao City, Zhejiang Province, so as to provide the basis for formulating weight management strategies for students.
Methods:
A census was conducted to investigate 72 primary schools and 23 middle schools students in Yuyao City in 2024, and data such as gender and age were collected through questionnaire surveys. Height and weight were measured to calculate the body mass index (BMI). Based on the BMI thresholds for different gender and age groups, overweight and obesity were determined.
Results:
Totally 75 082 individuals were surveyed, including 40 435 boys and 34 647 girls. Among them, 55 172 were primary school students and 19 910 were middle school students. A total of 8 677 overweight and 7 042 obese individuals were identified, with detection rates of 11.56% and 9.38%, respectively. The detection rates of overweight and obesity in boys were higher than in girls (14.01% vs. 8.69%, 11.58% vs. 6.81%, both P<0.05). The detection rate of obesity among students in rural schools was higher than students in urban schools (10.12% vs. 8.18%, P<0.05). The detection rates of overweight and obesity of students with non-local household registration were higher than students with local household registration (12.04% vs. 10.77%, 10.44% vs. 7.64%, both P<0.05). The detection rate of overweight showed an upward trend with age (P<0.05), while no significant age-related trend was observed for obesity (P>0.05).
Conclusions
The detection rates of overweight and obesity were notably high among primary and middle school students in Yuyao City. Boys, students in rural schools, students with non-local household registration, and older students are the key population.
5.Effect of carbon dioxide fractional laser combined with glucocorticoid on follicular helper T cells and B cell subsets in patients with advanced vitiligo
Lina ZHANG ; Chao LYU ; Zhixia FAN ; Xin WANG ; Juan SHEN ; Zhonglin ZHAO ; Jun′e FENG ; Guoying MIAO
Chinese Journal of Immunology 2025;41(5):1175-1181
Objective:To explore the effect of CO2 fractional laser combined with glucocorticoid(GC)on T follicular helper cell(Tfh)and B cell subsets in patients with advanced vitiligo(VL).Methods:Patients with advanced VL(128 cases)who were treated in Affiliated Hospital of Hebei University of Engineering from August 2019 to March 2022 were selected as research objects,and divided into group A(64 cases,CO2 fractional laser combined with GC)and group B(64 cases,CO2 fractional laser)according to different treatment methods,clinical efficacy and safety of the two groups were compared.Another healthy subjects(64 cases)were selected as control group.Collected clinical data of the three groups of subjects for comparative analysis.Randomized walking model was used to evaluate the effect of CO2 fractional laser combined with GC on immune function and inflammatory response in patients with advanced VL.Results:Compared with group B,group A had a higher clinical total effective rate(P<0.05)and a lower probability of adverse reactions(P<0.05).After treatment,compared with control group,proportion of Tfh2 and Tfh17 in group A had no significant difference(P>0.05),while proportion of Tfh2 and Tfh17 in group B still had significant difference(P<0.05);compared with group B,proportion of Tfh2 in group A after treatment was higher(P<0.05),while proportion of Tfh17 type was lower(P<0.05).After treat-ment,compared with control group,proportion of memory transformed B cell in group B was still significantly different(P<0.05).After treatment,compared with control group,levels of IgA,IgE and IgM in group B were still significantly different(P<0.05);compared with group B,level of IgM in group A was higher after treatment(P<0.05).After treatment,compared with control group,levels of IL-21 and IL-1β in group B were still significantly different(P<0.05);compared with group B,level of IL-10 in group A after treatment was higher(P<0.05),while levels of IL-21 and IL-1β were lower(P<0.05).Evaluation results of random walking model showed that the improvement of immune function and inflammatory reaction in group A was better than that in group B.Conclusion:CO2 fractional laser combined with GC can improve the immune function and inflammatory reaction of patients with advanced VL better after treatment,and the probability of adverse reactions is lower.
6.Mechanism of miR-518a-5p/HDAC6 axis in DNA oxidative damage in ovarian cancer SKOV3 cells
Ling ZHU ; Weili CAI ; Chao LIU ; Guoying XU ; Miao ZHANG ; Yunfeng YE
Chinese Journal of Comparative Medicine 2025;35(3):71-81
Objective To investigate the mechanism of the miR-518a-5p/histone deacetylase 6(HDAC6)axis in DNA oxidative damage in ovarian cancer(OC)SKOV3 cells.Methods Expression levels of miR-518a-5p and HDAC6 mRNA in OC tissues and in various cancer cells(A2780,SKOV3,CAOV3)were detected by qRT-PCR.SKOV3 cells were separated into Control,miR-NC,miR-518a-5p mimics,miR-518a-5p mimics+pcDNA-NC,and miR-518a-5p mimics+pc-HDAC6 groups.Cell proliferation and apoptosis were analyzed by colony-forming assay and Hoechst 33258 staining.Expression of phosphorylated histone H2AX(γ-H2AX)was detected by immunofluorescence assay and reactive oxygen species(ROS)were detected by flow cytometry.HDAC6,Bcl-2-associated X protein(Bax),and B-cell lymphoma-2(Bcl-2)protein expression were analyzed by Western blot.The regulatory relationship between miR-518a-5p and HDAC6 was analyzed by dual luciferase assay.The effect and mechanism of miR-518a-5p on oxidative DNA damage in OC cells were studied in a xenotransplantation tumor model.Results miR-518a-5p expression was decreased and HDAC6 expression was increased in OC tissues and A2780,SKOV3,and CAOV3 cells(P<0.001).Expression levels of miR-518a-5p were lowest and expression levels of HDAC6 were highest in SKOV3 cells,and SKOV3 cells were therefore selected for subsequent experiments.miR-518a-5p expression,apoptosis rate,number of γ-H2AX-positive cells,relative ROS fluorescence intensity,and expression of Bax were all higher in the miR-518a-5p mimics group compared with the miR-NC group,while HDAC6 mRNA and protein expression,Bcl-2 expression,and colony-formation number were all lower(P<0.001).HDAC6 mRNA and protein expression,colony-formation number,and expression of Bcl-2 were higher in the miR-518a-5p mimics+pc-HDAC6 group compared with the miR-518a-5p mimics+pcDNA-NC group,and the apoptosis rate,number of γ-H2AX-positive cells,relative ROS fluorescence intensity,and expression of Bax were all lower(P<0.001).HDAC6 had a targeted regulatory relationship with miR-518a-5p.Overexpression of miR-518a-5p decreased tumor volume,weight,and HDAC6 protein expression in tumor tissues,and increased γ-H2AX expression in vivo(P<0.001).Upregulation of HDAC6 expression by overexpression of miR-518a-5p increased graft tumor volume,weight,and HDAC6 protein expression and decreased γ-H2AX-positive expression(P<0.05).Conclusions miR-518a-5p expression is reduced and HDAC6 expression is increased in OC tissues and cells.Overexpression of miR-518a-5p can induce oxidative DNA damage in SKOV3 cells by inhibiting HDAC6 expression,thereby inhibiting cell proliferation and promoting cell apoptosis.
7.Feasibility analysis of laparoscopic total hysterectomy in patients with obliterated rectouterine pouch
Erniao LIU ; Fei MIAO ; Yingfang ZHOU ; Yan HUANG ; Lei ZHANG ; Chao PENG
Chinese Journal of Obstetrics and Gynecology 2025;60(5):355-362
Objective:To analysis the safety and feasibility of laparoscopic total hysterectomy in patients with obliterated rectouterine pouch.Methods:A retrospective analysis was conducted on 197 patients who underwent laparoscopic total hysterectomy at Peking University First Hospital, from July 2022 to July 2024. According to intraoperative observations, 197 patients were categorized into two groups: 58 cases with completely obliterated rectouterine pouch (obliterated group) and 139 cases with non-obliterated pouch (control group). General clinical characteristics, perioperative outcomes and 3-month follow-up after surgery were compared between two groups.Results:(1) Baseline characteristics: the age of the obliterated group was (45.7±3.7) years, and the control group was (48.0±3.8) years ( P<0.001). Primary complaint: dysmenorrhea prevalence was 53.4% (31/58) in the obliterated group, and was 35.3% (49/139; P<0.05) in the control group. Surgical indications: the proportion of adenomyosis and ovarian endometrioma was 77.6% (45/58) in the obliterated group, and was 51.1% (71/139; P<0.001) in the control group. No significant differences in body mass index, pelvic surgical history, or preoperative medication between the two groups (all P>0.05). (2) Intraoperative outcomes: the obliterated group demonstrated significantly higher rates of surgical interventions compared to the control group. Superficial endometriosis resection was performed in 91.4% (53/58) of the obliterated group versus 33.8% (47/139) in the control group ( P<0.001). Deep infiltrating endometriosis excision was required in 82.8% (48/58) of the obliterated group, contrasting sharply with 10.1% (14/139) in the control group ( P<0.001). Ureterolysis procedures were similarly elevated in the obliterated group (77.6%, 45/58) compared to the control group (7.9%, 11/139; P<0.001). Operative metrics revealed substantial intergroup disparities: the obliterated group exhibited a median surgical duration of 149.0 minutes (interquartile range: 114.0, 180.0 minutes), significantly prolonged relative to the control group′s 91.0 minutes (77.0, 107.0 minutes; P<0.001). Estimated blood loss followed a parallel pattern, with median volumes of 50.0 ml (20.0, 100.0 ml) in the obliterated group versus 20.0 ml (10.0, 20.0 ml) in the control group ( P<0.001). (3) Postoperative outcomes: the uterine weight of the obliterated group was 200.0 g (132.5, 260.0 g), and the control group was 240.0 g (180.0, 336.0 g; P<0.05). Hospital stay was prolonged in the obliterated group compared with the control group [7.0 days (6.0, 8.3 days) vs 6.0 days (5.0, 7.0 days); P<0.001]. The incidences of postoperative fever in the obliterated group and the control group were 20.7% (12/58) vs 12.2% (17/139; P>0.05). The incidences of minor complications in the obliterated group and the control group were 3.4% (2/58) vs 0 (0/139; P>0.05). No major complications (intraoperative hemorrhage, transfusion, visceral injury, conversion to laparotomy, or thromboembolism) occurred. (4) Follow up at 3 months after surgery: three months postoperatively, all patients underwent outpatient follow-up visits, during which symptoms such as pain were alleviated. Gynecological ultrasound and pelvic examinations were performed, and the vaginal stump exhibited good healing. Conclusion:Laparoscopic total hysterectomy performed by experienced minimally invasive gynecological specialists is safe and feasible for patients with obliterated rectouterine pouch.
8.Investigation of the prevalence of progestogen drug contraindications in patients with endometriosis and adenomyosis
Fei MIAO ; Erniao LIU ; Yingfang ZHOU ; Yan HUANG ; Lei ZHANG ; Chao PENG
Chinese Journal of Obstetrics and Gynecology 2025;60(7):534-540
Objective:To discuss the prevalence of contraindications to progestogens in patients with endometriosis and adenomyosis.Methods:Women of childbearing age aged 15-49 years, except for menopausal and malignant tumors, were enrolled in Peking University First Hospital from April to August 2024 for laparoscopic or open surgery, regardless of whether they had used or would be using progestogen drugs. The patients were divided into two groups: the case group was patients with pathologically confirmed endometriosis and adenomyosis, and the control group was patients with other benign gynecological diseases. The patient′s medical history and clinical data were collected.Results:A total of 745 patients were enrolled, including 362 cases (48.6%, 362/745) in the case group, 383 cases (51.4%, 383/745) in the control group; 61 patients with contraindications to progestogens, including 32 cases (8.8%, 32/362) in the case group, and 29 cases (7.6%, 29/383) in the control group ( χ2=0.398, P>0.05). Among them, there were 33 cases of liver disease, all of which were liver tumor (hepatic hemangioma only), including 18 cases (5.0%, 18/362) in the case group and 15 cases (3.9%, 15/383) in the control group. There were 11 cases of sex hormone-dependent or related tumors (breast cancer only), including 4 cases (1.1%, 4/362) in the case group and 7 cases (1.8%, 7/383) in the control group. There were 10 cases of vascular disease, including 4 cases of diabetic vascular disease, including 2 cases (0.6%, 2/362) in the case group and 2 cases (0.5%, 2/383) in the control group. There were 3 cases of venous thrombosis, including 2 cases (0.6%, 2/362) in the case group and 1 case (0.3%, 1/383) in the control group. There were 3 cases of cardiovascular and cerebrovascular diseases, including 1 case in the case group (0.3%, 1/362) and 2 cases (0.5%, 2/383) in the control group. There were 7 cases of renal insufficiency, including 3 cases of chronic nephritis complicated with renal insufficiency [including 2 cases (0.6%, 2/362) in the case group and 1 case (0.3%, 1/383) in the control group]; two cases of membranous nephropathy complicated with renal insufficiency were in the case group (0.6%, 2/362); one case of diabetic nephropathy complicated with renal insufficiency was in the control group (0.3%, 1/383); one case of uremia was in the case group (0.3%, 1/362). There were no statistically significant differences between the two groups (all P>0.05). Conclusions:There is no difference in the contraindications of progestogens in patients with endometriosis and adenomyosis compared with patients with other gynecological benign diseases. Liver tumors are more common in both two groups.
9.Effect of carbon dioxide fractional laser combined with glucocorticoid on follicular helper T cells and B cell subsets in patients with advanced vitiligo
Lina ZHANG ; Chao LYU ; Zhixia FAN ; Xin WANG ; Juan SHEN ; Zhonglin ZHAO ; Jun′e FENG ; Guoying MIAO
Chinese Journal of Immunology 2025;41(5):1175-1181
Objective:To explore the effect of CO2 fractional laser combined with glucocorticoid(GC)on T follicular helper cell(Tfh)and B cell subsets in patients with advanced vitiligo(VL).Methods:Patients with advanced VL(128 cases)who were treated in Affiliated Hospital of Hebei University of Engineering from August 2019 to March 2022 were selected as research objects,and divided into group A(64 cases,CO2 fractional laser combined with GC)and group B(64 cases,CO2 fractional laser)according to different treatment methods,clinical efficacy and safety of the two groups were compared.Another healthy subjects(64 cases)were selected as control group.Collected clinical data of the three groups of subjects for comparative analysis.Randomized walking model was used to evaluate the effect of CO2 fractional laser combined with GC on immune function and inflammatory response in patients with advanced VL.Results:Compared with group B,group A had a higher clinical total effective rate(P<0.05)and a lower probability of adverse reactions(P<0.05).After treatment,compared with control group,proportion of Tfh2 and Tfh17 in group A had no significant difference(P>0.05),while proportion of Tfh2 and Tfh17 in group B still had significant difference(P<0.05);compared with group B,proportion of Tfh2 in group A after treatment was higher(P<0.05),while proportion of Tfh17 type was lower(P<0.05).After treat-ment,compared with control group,proportion of memory transformed B cell in group B was still significantly different(P<0.05).After treatment,compared with control group,levels of IgA,IgE and IgM in group B were still significantly different(P<0.05);compared with group B,level of IgM in group A was higher after treatment(P<0.05).After treatment,compared with control group,levels of IL-21 and IL-1β in group B were still significantly different(P<0.05);compared with group B,level of IL-10 in group A after treatment was higher(P<0.05),while levels of IL-21 and IL-1β were lower(P<0.05).Evaluation results of random walking model showed that the improvement of immune function and inflammatory reaction in group A was better than that in group B.Conclusion:CO2 fractional laser combined with GC can improve the immune function and inflammatory reaction of patients with advanced VL better after treatment,and the probability of adverse reactions is lower.
10.Mechanism of miR-518a-5p/HDAC6 axis in DNA oxidative damage in ovarian cancer SKOV3 cells
Ling ZHU ; Weili CAI ; Chao LIU ; Guoying XU ; Miao ZHANG ; Yunfeng YE
Chinese Journal of Comparative Medicine 2025;35(3):71-81
Objective To investigate the mechanism of the miR-518a-5p/histone deacetylase 6(HDAC6)axis in DNA oxidative damage in ovarian cancer(OC)SKOV3 cells.Methods Expression levels of miR-518a-5p and HDAC6 mRNA in OC tissues and in various cancer cells(A2780,SKOV3,CAOV3)were detected by qRT-PCR.SKOV3 cells were separated into Control,miR-NC,miR-518a-5p mimics,miR-518a-5p mimics+pcDNA-NC,and miR-518a-5p mimics+pc-HDAC6 groups.Cell proliferation and apoptosis were analyzed by colony-forming assay and Hoechst 33258 staining.Expression of phosphorylated histone H2AX(γ-H2AX)was detected by immunofluorescence assay and reactive oxygen species(ROS)were detected by flow cytometry.HDAC6,Bcl-2-associated X protein(Bax),and B-cell lymphoma-2(Bcl-2)protein expression were analyzed by Western blot.The regulatory relationship between miR-518a-5p and HDAC6 was analyzed by dual luciferase assay.The effect and mechanism of miR-518a-5p on oxidative DNA damage in OC cells were studied in a xenotransplantation tumor model.Results miR-518a-5p expression was decreased and HDAC6 expression was increased in OC tissues and A2780,SKOV3,and CAOV3 cells(P<0.001).Expression levels of miR-518a-5p were lowest and expression levels of HDAC6 were highest in SKOV3 cells,and SKOV3 cells were therefore selected for subsequent experiments.miR-518a-5p expression,apoptosis rate,number of γ-H2AX-positive cells,relative ROS fluorescence intensity,and expression of Bax were all higher in the miR-518a-5p mimics group compared with the miR-NC group,while HDAC6 mRNA and protein expression,Bcl-2 expression,and colony-formation number were all lower(P<0.001).HDAC6 mRNA and protein expression,colony-formation number,and expression of Bcl-2 were higher in the miR-518a-5p mimics+pc-HDAC6 group compared with the miR-518a-5p mimics+pcDNA-NC group,and the apoptosis rate,number of γ-H2AX-positive cells,relative ROS fluorescence intensity,and expression of Bax were all lower(P<0.001).HDAC6 had a targeted regulatory relationship with miR-518a-5p.Overexpression of miR-518a-5p decreased tumor volume,weight,and HDAC6 protein expression in tumor tissues,and increased γ-H2AX expression in vivo(P<0.001).Upregulation of HDAC6 expression by overexpression of miR-518a-5p increased graft tumor volume,weight,and HDAC6 protein expression and decreased γ-H2AX-positive expression(P<0.05).Conclusions miR-518a-5p expression is reduced and HDAC6 expression is increased in OC tissues and cells.Overexpression of miR-518a-5p can induce oxidative DNA damage in SKOV3 cells by inhibiting HDAC6 expression,thereby inhibiting cell proliferation and promoting cell apoptosis.


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