1.Effects of ursolic acid on iron death in endometrial stromal cells via JAK2/STAT3 signaling pathway
Cong WANG ; Guixiang MEN ; Shuai SHAO ; Lina MA
Chinese Journal of Endocrine Surgery 2023;17(3):317-322
Objective:To investigate the effects of ursolic acid (UA) on proliferation, migration and iron death of ectopic endometrial stromal cells (EESCs) and its mechanism.Methods:Mouse model of endometriosis was established and the primary EESCs were isolated. The cells were treated with UA at different concentrations (0, 2.5, 5, 10, 20, 40, 50, 80, 100, 200 μmol/L). The cells were divided into Control group (normal culture), 2.5 μmol/L UA group (2.5 μmol/L UA treatment), 5.0 μmol/L UA group (5.0 μmol/L UA treatment), 10.0 μmol/L UA group (10 μmol/L UA treatment), and UA+DUSP19 group (10 μmol/L UA+50 μmol/L JAK2/STAT3 signal pathway activator DUSP19 treatment). Cell survival rate was detected by CCK-8 method. Cell proliferation was detected by plate cloning method. Transwell chamber assay was used to detect cell migration. The levels of Fe 2+ and the contents of malondialdehyde (MDA), reactive oxygen species (ROS) and superoxide dismutase (SOD) were detected by kit. Protein expression levels of Ki67, PCNA, CyclinD1, p-JAK2, p-STAT3, JAK2 and STAT3 were detected by western blot. Results:The number of clones in Control, 2.5 μmol/L UA, 5.0 μmol/L UA and 10.0 μmol/L UA groups were as follows: 152.22±15.47, 121.22±11.54, 92.00±5.54, 66.44±6.88; Ki67 protein expression was 1.08±0.10, 0.73±0.07, 0.61±0.06, 0.45±0.02, respectively; The expression of PCNA protein was 0.85±0.07, 0.64±0.05, 0.41±0.03, 0.31±0.05, respectively; CyclinD1 protein expression levels were 0.98±0.11, 0.65±0.06, 0.51±0.05, 0.42±0.07, respectively. The migration numbers were 92.78±6.27, 62.22±2.20, 50.22±4.59 and 39.11±4.33, respectively; Fe 2+ levels were (1.06±0.07) μmol/g, (1.21±0.11) μmol/g, (1.33±0.08) μmol/g, (1.47±0.09) μmol/g, respectively; MDA content was (0.48±0.06) μmol/g, (0.65±0.07) μmol/g, (0.85±0.08) μmol/g, (1.03±0.11) μmol/g, respectively; ROS contents were (19.85±1.21) %, (24.83±2.79) %, (29.04±1.86) %, (33.87±2.45) %, respectively; SOD content were (36.41±3.56) U/mg, (31.03±2.81) U/mg, (25.63±2.84) U/mg, (19.62±1.67) U/mg, respectively; p-JAK2 protein expression was 0.85±0.10, 0.75±0.06, 0.53±0.05, 0.31±0.03, respectively; p-STAT3 protein expression was 1.08±0.11, 0.79±0.06, 0.63±0.07, 0.42±0.03, respectively. The p-JAK2 protein content in UA group and UA+DUSP19 group was 0.38±0.05 and 0.75±0.08, respectively; p-STAT3 protein expression was 0.46±0.04 and 0.80±0.03, respectively; The cell survival rates were (52.55±2.44) % and (82.18±4.72) %, respectively; Fe 2+ levels were (1.57±0.06) μmol/g and (1.21±0.13) μmol/g, respectively. The differences in the above indicators between the Control group and the 2.5 μmol/L UA group, 5.0 μmol/L UA group and 10.0 μmol/L UA group were statistically significant ( P<0.05). There were statistically significant differences among 2.5 μmol/L UA group, 5.0 μmol/L UA group and 10.0 μmol/L UA group ( P<0.05). There were statistically significant differences in p-JAK2, p-STAT3, cell survival rate and Fe 2+ levels between UA group and UA+DUSP19 group ( P<0.05) . Conclusion:Ursolic acid can inhibit the proliferation and migration of EESCs cells and induce iron death by regulating JAK2/STAT3 signaling pathway, thus playing a protective role in endometriosis.
2.Effect of single nucleotide variation of OPG gene on osteoporosis in patients with gestational diabetes mellitus
Lina MA ; Cong WANG ; Guixiang MEN ; Yongheng ZHAO ; Shuai SHAO
Chinese Journal of Endocrine Surgery 2023;17(2):209-213
Objective:To investigate the effect of single nucleotide variation of osteoprotegerin (OPG) gene on the occurrence of osteoporosis (OP) in patients with gestational diabetes mellitus (GDM) .Methods:From Apr. 2018 to Apr. 2022, 276 pregnant women with GDM who underwent prenatal examination and gave birth in Linyi People’s Hospital were collected for analysis, general data were collected and bone mineral density was tested. According to the bone mineral density test results, they were divided into normal group and OP group. The OPG genotype was tested, and the general information, OPG genotype and allele frequency of the two groups were compared. The differences in bone mineral density among different genotypes of OPG were compared, and the genotypes affecting the risk of OP in GDM patients were analyzed.Results:There was no significant difference in the general data of the two groups of patients (all P>0.05). The allelic distribution of the rs3134069 and rs2073618 loci of the OPG gene in the two groups of patients conformed to the Hardy-Weinberg equilibrium law (all P>0.05). There was a statistically significant difference in the frequency of the AC genotype at rs3134069 between the two groups ( χ2=7.75, P=0.005). Taking patients with the AA genotype as a reference, patients with the AC genotype had a lower risk of developing OP ( OR=0.15, 95% CI: 0.03-0.59). There was a statistically significant difference in the frequency of CC genotype at rs2073618 between the two groups ( χ2=11.30, P=0.001). Taking patients with GG genotype as a reference, patients with CC genotype had a higher risk of developing OP ( OR=7.42, 95% CI: 2.19-27.18). Comparing rs3134069 and rs2073618 loci, there was no significant difference in bone mineral density at each part of the three genotypes (all P>0.05). The multivariate Logistic regression model showed that the AC genotype of rs3134069 ( OR=0.18, 95% CI: 0.03-0.70, P=0.029) was a protective factor for the induction of OP, while GC genotype of rs2073618 ( OR=6.86, 95% CI: 1.57-27.15, P=0.007) were the risk factors for OP in GDM patients. Conclusion:The CC genotype of rs2073618 is significantly positively correlated with the susceptibility to OP in GDM patients.
3.Prediction of clinical efficacy of breast cancer neoadjuvant chemotherapy using shear wave elastography
Dianxia MEN ; Chunsong KANG ; Jiping XUE ; Xiaoyan KANG ; Shuai LI ; Lili SU
Chinese Journal of Ultrasonography 2017;26(9):781-786
Objective To investigate the value of shear wave elastography(SWE)to early predict clinical efficacy of breast cancer neoadjuvant chemotherapy(NAC).Methods Fifty-three patients(55 lesions)with breast cancer who underwent NAC were enrolled in this study.SWE was performed at baseline and after the end of 2,4,6 cycles of NAC.According to the postoperative pathological results,the lesions were divided into major pathologic response group and minor pathologic response group.The maximum diameter of the lesions between the baseline and the end of the last cycle of the two groups were measured and the maximum diameter changes(ΔDmax)of the two groups were compared.The maximum elastic modulus(Emax)and the maximum elastic modulus changes(ΔEmax)of the two groups were measured and compared at baseline and after the end of 2,4,6 cycles of NAC.The ROC curves of ΔEmax at the end of 2 and 4 cycles were used to evaluate the predictive value of NAC response.Results The ΔDmax of the major pathologic response group[(64.82±21.06)%]was higher than that of the minor pathologic response group[(26.49±26.1 1)%,P <0.001].With the prolongation of NAC,the values of Emax were significantly decreased in the two groups,but the decreasing degrees were different.The Emax was significantly decreased at the end of 2 cycles in the major pathologic response group(P <0.05),however, at the end of 4 cycles in minor pathologic response group(P <0.05).The value of ΔEmax in the major pathologic response group was higher than that in the minor pathologic response group in every cycle significantly(all P <0.05).The ΔEmax threshold of the 2 cycles for predicting the NAC response was 26.1%(81.8% of sensitivity,75% of specificity),while the threshold of the 4 cycles was 35.5%(87.9%of sensitivity,83.3% of specificity),there was no significant difference between the areas under the two curves(P =0.264).Conclusions The value of ΔEmax in breast cancer lesions can predict the efficacy of NAC early.SWE can provide a valuable complement for two-dimensional ultrasound in the evaluation of NAC efficacy in breast cancer.
4.Expression and clinical significance of runt-related transcription factor 3 in children with bronchiolitis.
Shuai MEN ; Yan-Yan YU ; Yu-Hong ZHANG ; Yi-Fen WANG ; Qian QIAN ; Wei LI ; Chuang YIN
Chinese Journal of Contemporary Pediatrics 2019;21(10):1005-1011
OBJECTIVE:
To study the mRNA level of runt-related transcription factor 3 (RUNX3) in children with bronchiolitis and its clinical significance in bronchiolitis.
METHODS:
A total of 54 young children with bronchiolitis were enrolled as the bronchiolitis group, among whom 28 with atopic constitution were enrolled in the atopic bronchiolitis group and 26 with non-atopic constitution were enrolled in the non-atopic bronchiolitis group. A total of 48 healthy young children were enrolled as the healthy control group, among whom 24 with atopic constitution were enrolled in the atopic healthy control group and 24 with non-atopic constitution were enrolled in the non-atopic healthy control group. Quantitative real-time PCR was used to measure the mRNA level of RUNX3 in peripheral blood mononuclear cells. ELISA was used to measure the serum levels of interleukin-4 (IL-4) and interferon gamma (IFN-γ).
RESULTS:
The bronchiolitis group had a significantly lower mRNA level of RUNX3 than the healthy control group, and the atopic bronchiolitis group had a significantly lower mRNA level of RUNX3 than the non-atopic bronchiolitis, atopic healthy control, and non-atopic healthy control groups (P<0.05). The bronchiolitis group had a significantly higher serum level of IL-4 than the healthy control group, and the atopic bronchiolitis group had a significantly higher serum level of IL-4 than the non-atopic healthy control group (P<0.05). The bronchiolitis group had a significantly lower serum level of IFN-γ than the healthy control group, and the atopic bronchiolitis group had a significantly lower serum level of IFN-γ than the non-atopic bronchiolitis, atopic healthy control, and non-atopic healthy control groups (P<0.05). The correlation analysis showed that the mRNA level of RUNX3 was negatively correlated with the serum level of IL-4 and was positively correlated with the serum level of IFN-γ (P<0.05).
CONCLUSIONS
Measurement of RUNX3 gene expression in peripheral blood mononuclear cells has a certain value in identifying children with atopic constitution at high risk of asthma among children with bronchiolitis.
Asthma
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Bronchiolitis
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Child
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Child, Preschool
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Core Binding Factor Alpha 3 Subunit
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genetics
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Humans
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Interferon-gamma
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Leukocytes, Mononuclear