Objective : To explore the expression of prohibitin2(PHB2) in breast cancer and its effect on the biological behaviors of tumor cells. Methods : Immunohistochemistry was used to detect the expression of PHB2 protein in breast cancer tissues and its relationship with clinicopathologic features. Breast cancer stable transient cell lines were constructed with knockdown and overexpression ofPHB2, respectively. The effects of PHB2 on cell proliferation, migration and invasion ability were detected by clone formation assay, scratch assay and Transwell assay. Western blot(WB) was used to detect the effects of PHB2 on the expression of epithelial-mesenchymal transition(EMT)-related markers, including E-cadherin, N-cadherin, Snail family transcriptional repressor 1(Snail) protein, Vimentin, and Claudin-1. The effect of PHB2 on tumorigenicityin vivowas detected by subcutaneous tumor formation assay in nude mice. Results: The result of immunohistochemical showed that PHB2 was highly expressed in breast cancer and the expression of PHB2 was significantly positive correlated with tumor size, human epidermal growth factor receptor-2(HER-2) status and proliferation index Ki-67 levels(P<0.05). Clone formation assay, scratch assay and Transwell assay revealed that knockdown ofPBH2significantly inhibited the proliferation, migration and invasion ability of breast cancer cells(P<0.01), while the overexpression ofPHB2significantly promoted cell proliferation, migration and invasion(P<0.01). The result of subcutaneous tumor formation experiment in nude mice revealed a significant decrease in tumor volume and weight in knockdownPHB2mice(P<0.000 1), whilePHB2overexpression tumors significantly increased in volume and weight(P<0.001).WB assay showed that the protein expression of epithelial marker E-cadherin increased, while the expressions of mesenchymal markers N-cadherin, Snail and Vimentin decreased significantly afterPHB2knockdown with them in control cells(P<0.01). The expression of Claudin-1 decreased, while the expressions of N-cadherin, Snail and Vimentin increased significantly inPHB2overexpression cells(P<0.05). Conclusion PHB2 is highly expressed in breast cancer and promotes multiple malignant biological behaviors in tumor cells, suggesting PHB2 may be a potential target for breast cancer diagnosis and treatment.

BACKGROUND:Eucommia ulmoides has a certain osteogenic effect,which can promote the proliferation and differentiation of osteoblasts.However,it is unclear whether Eucommia ulmoides has effects on alveolar bone formation and Wnt/β-Catenin signaling pathway. OBJECTIVE:To investigate the mechanism by which Eucommia ulmoides promotes alveolar bone formation in ovariectomized rats based on the Wnt/β-Catenin signaling pathway. METHODS:Sixty female Sprague-Dawley rats were selected and randomly divided into five groups:blank control group,sham-operation group,model group,low-dose group Eucommia ulmoides group,and high-dose Eucommia ulmoides group,with twelve rats in each group.Osteoporosis animal models were constructed by bilateral oophorectomy in the model group and the low-dose and high-dose Eucommia ulmoides groups.The sham-operation group underwent the same method to remove adipose tissue of equal mass around the bilateral ovaries.Three months after surgery,the low-and high-dose Eucommia ulmoides groups were given 2.1 g/kg/d and 4.2 g/kg/d Eucommia ulmoides by gavage,respectively.The sham-operation group and model group were given the same amount of physiological saline by gavage.After 12 weeks of drug intervention,the changes in alveolar bone mass of rats in each group were observed through Micro-CT;hematoxylin-eosin staining was used to observe the pathological structural changes of alveolar bone in rats;enzyme linked immunosorbent assay was used to detect the expression levels of alkaline phosphatase and osteocalcin in the serum of rats;western blot was used to detect the expression levels of β-Catenin and Frizzled9 receptor proteins in the alveolar bone of rats;and real-time fluorescence quantitative PCR was used to detect the expression of osteocalcin,Runt-related transcription factor 2(Runx2),alkaline phosphatase,β-catenin,and frizzled9 mRNAs in alveolar bone tissues of rats. RESULTS AND CONCLUSION:Compared with the blank control group,bone volume fraction,trabecular number,trabecular thickness,and bone mineral density were reduced in the model group(P＜0.05),and trabecular separation was elevated(P＜0.05).Pathological observation showed that the arrangement of trabeculae was disordered and irregular,the trabeculae were thinned or broken,and the marrow cavity was enlarged in the model group,with a significant reduction in bone volume;the level of alkaline phosphatase in the serum was increased(P＜0.05),and the level of osteocalcin was decreased(P＜0.05);mRNA expression of alkaline phosphatase,osteocalcin,Runx2,β-catenin,and frizzled9 were decreased(P＜0.05);protein expression of β-Catenin and Frizzled9 was decreased(P＜0.05).Compared with the model group,the low-and high-dose Eucommia ulmoides groups showed an increase in bone volume fraction,trabecular number,trabecular thickness,and bone mineral density(P＜0.05)and a decrease in trabecular separation(P＜0.05).In the low-and high-dose Eucommia ulmoides groups,bone trabeculae were slightly aligned and thickened,with a significant increase in bone mass.Compared with the model group,the serum level of alkaline phosphatase was reduced(P＜0.05)and the serum level of osteocalcin was elevated(P＜0.05)in the low-and high-dose Eucommia ulmoides groups.Compared with the model group,the mRNA expression of alkaline phosphatase,osteocalcin,Runx2,β-catenin,and frizzled9 were increased in the low-and high-dose Eucommia ulmoides groups(P＜0.05).Compared with the model group,the protein expression of Frizzled9 was increased in the low-dose Eucommia ulmoides group(P＜0.05),while the protein expression of β-Catenin and Frizzled9 was increased in the high-dose Eucommia ulmoides group(P＜0.05).Compared with the low-dose Eucommia ulmoides group,the high-dose Eucommia ulmoides group had a more significant improvement in the above indexes.To conclude,Eucommia ulmoides can effectively promote the alveolar bone formation,and its mechanism of action might be related to the activation of the Wnt/β-catenin signaling pathway.

BACKGROUND:Rev-erbα is involved in the regulation of inflammation,but pharmacological activation of Rev-erbα increases the risk for cardiovascular diseases.To reduce the relevant risk,an exploration on SR9009,a Rev-erbα agonist,combined with other drugs to relieve inflammation in skeletal myoblasts was conducted,laying the theoretical foundation for the treatment of inflammation-associated skeletal muscle atrophy. OBJECTIVE:To investigate the relationship of SR9009,indolepropionic acid and nuclear factor-κB signaling pathways in lipopolysaccharide-induced C2C12 myoblasts. METHODS:(1)C2C12 myoblasts were induced to differentiate in the presence of lipopolysaccharide(1 μg/mL).RNA-seq and KEGG pathway analysis were used to study signaling pathways.(2)C2C12 myoblast viability was assessed using the cell counting kit-8 assay to determine optimal concentrations of indolepropionic acid.Subsequently,cells were categorized into control group,lipopolysaccharide(1 μg/mL)group,SR9009(10 μmol/L)+lipopolysaccharide group,indolepropionic acid(80μmol/L)+lipopolysaccharide group,and SR9009+indolepropionic acid+lipopolysaccharide group.ELISA was employed to measure protein expression levels of interleukin-6 in the cultured supernatant.Real-time quantitative PCR were employed to measure mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Western blot assay were employed to measure protein expression levels of NF-κB p65 and p-NF-κB p65.(3)After Rev-erbα was knocked down by siRNA,knockdown efficiency was assessed by RT-qPCR.And mRNA levels of interleukin-6 and tumor necrosis factor α were also measured. RESULTS AND CONCLUSION:Compared with the blank control group,lipopolysaccharide time-dependently inhibited myofibroblast fusion to form myotubes,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were elevated,and the level of interleukin-6 in the cell supernatant was significantly increased.The results of KEGG pathway showed that the nuclear factor-κB signaling pathway was activated by lipopolysaccharide.Indolepropionic acid exhibited significant suppression of C2C12 myoblasts viability when its concentration exceeded 80 μmol/L.Indolepropionic acid and SR9009 inhibited the activation of NF-κB signaling pathway,thereby played an anti-inflammatory role,and suppressed the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.Compared with the lipopolysaccharide group,the ratio of p-NF-κB p65/NF-κB p65 protein expression were downregulated.SR9009 combined with indolepropionic acid notably reduced lipopolysaccharide-induced inflammation,further downregulated the mRNA expression levels of interleukin-6,tumor necrosis factor α,TLR4 and CD14.The ratio of p-NF-κB p65/NF-κB p65 protein expression was significantly lower than that in the SR9009+lipopolysaccharide group or indolepropionic acid+lipopolysaccharide group.Rev-erbα increases time-dependently with lipopolysaccharide induction.The knockdown efficiency of Rev-erbα by siRNA reached over 58%,and lipopolysaccharide was added after Rev-erbα was successfully knocked down.Compared with the lipopolysaccharide group,the mRNA expression levels of interleukin-6 and tumor necrosis factor α were significantly up-regulated.These results conclude that Rev-erbα may act as a promising pharmacological target to reduce inflammation.SR9009 targeted activation of Rev-erbα combined with indolepropionic acid significantly inhibits the nuclear factor-κB signaling pathway and attenuates the inflammatory response of C2C12 myofibroblasts.Moreover,the combined anti-inflammatory effect is superior to that of the intervention alone.

Objective:To investigate the regulatory effect of electroacupuncture on visual cortex plasticity in adult amblyopic mice and its mechanism.Methods:Forty-eight SPF male healthy 3-week-old Kunming mice were randomly divided into a normal control group, a monocular form deprivation (MD) group, and an electroacupuncture intervention group by the random number table method, with 16 mice in each group.Except for the normal control group, mice in the other groups had their right eyelids sutured for two weeks to establish an adult MD amblyopia model.The electroacupuncture intervention group received electroacupuncture stimulation at three acupoints, Taiyang (EX-HN5), Jingming (BL1), and Fengchi (GB20) for four weeks at five weeks of age.The subjective visual function (paw probing success rate) of each group of mice was measured at five and nine weeks of age, respectively.At nine weeks of age, the changes in flash-visual evoked potential (F-VEP) of mice in each group was detected.The expression of plasticity related proteins synaptophysin (SYP), synaptic protein 1 (SYN1), neurite overgrowth inhibitor A (Nogo-A), and Nogo receptor (NgR) proteins in the contralateral cortex of the deprived mouse eyes was detected by Western blot.Expression of early growth response gene 1 (Egr-1) in the visual cortex of mice was detected by immunohistochemical staining.This study was approved by the Animal Ethics Committee of Hunan Children's Hospital (No.HCHDWLL-2022-17), and the management and use of animals were in accordance with the Laboratory Animal Management and Use Guide of Hunan Children's Hospital.Results:At nine weeks of age, the success rates of paw probing in the normal control group, MD group, and electroacupuncture intervention group were (71.69±10.60)%, (25.54±10.09)%, and (58.25±8.39)%, respectively, with a statistically significant overall difference ( F=5.987, P=0.006).Among them, the success rate of paw probing was significantly lower in the MD group than in the normal control group, and the electroacupuncture intervention group was significantly higher than in the MD group (both P<0.05).There was a significant overall difference in P2 wave amplitude in F-VEP examination among different groups of mice ( F=63.710, P<0.001), with lower P2 wave amplitude in the deprived eye of the MD group than in the normal control group and the electroacupuncture intervention group, and the differences were statistically significant (both P<0.001).There were significant differences in the expression levels of SYP and SYN1 proteins in the contralateral cortex of the deprived mouse eyes ( F=5.451, 3.871; both P<0.05).The relative expression levels of SYP and SYN1 proteins were significantly lower in the MD group than in the normal control group and electroacupuncture intervention group (all P<0.05).There were significant differences in levels of Nogo-A and NgR proteins in the contralateral cortex of the deprived mouse eyes ( F=4.188, 3.942, both P<0.05).The relative expression levels of Nogo-A and NgR proteins were significantly higher in the MD group than in the normal control group and the electroacupuncture intervention group (all P<0.05).The immunohistochemical staining results showed that compared with the normal control group, the MD group mice had a decrease in the expression of Egr-1 in the contralateral cortical neurons of the deprived eye, and the brown neuron protrusions were indistinguishable.Compared with the MD group, the electroacupuncture intervention group showed significant positive cell expression in the contralateral cortical area of the deprived eye, but the expression intensity was weaker than that of the normal control group. Conclusions:Electroacupuncture treatment of Taiyang (EX-HN5), Jingming (BL1), and Fengchi (GB21) acupoints can reactivate the plasticity of the visual cortex in adult amblyopic mice and improve their visual function.The mechanism may be related to the regulation of Nogo-A/NgR signaling pathway.

Objective To construct a mammography image classification assistant system suitable for Chinese population,and explore the potential of artificial intelligence technology to assist early screening of breast cancer in China.Methods Curated breast imaging subset of digital database for screening mammography(CBIS-DDSM),Mammographic image analysis society database(MIAS)and other international open datasets were used to conduct model training respectively in order to reproduce the mainstream in-depth learning methods in the current literature.The model was also tested on the Chinese breast mammography database(CBMD)provided by Huajiao Technology Co.,Ltd,and the performance was compared.Aiming at the problem that the Chinese population data are not ideal in the performance test of the open dataset training model,an optimization strategy based on the sliding window adjustment mechanism was implemented in combination with the characteristics of Chinese population data.Then a two-stage migration learning method was designed to improve the overall performance of the model,and then development of our system was carried out.Results With the sliding window adjustment mechanism and the CBMD training model after two-stage transfer learning,the accuracy of our developed system was improved from 0.50 of the open datasets to 0.80,precision from 0.54 to 0.82,sensitivity from 0.52 to 0.80,F1 value from 0.52 to 0.80,and AUC value from 0.51 to 0.89 based on the Chinese population dataset as the test set.Conclusion Through the introduction of sliding window adjustment mechanism and two-stage migration learning strategy,the performance of the breast molybdenum target image classification model has been significantly improved in the Chinese population dataset,and our system primarily achieves the purpose of assisting the classification of breast molybdenum target images for the Chinese population.

Clinical chemotherapy for prostate cancer is still compromised by high treatment thresholds and severe off-target toxicity of drugs. Given the limited progress in improving therapeutic outcomes and reducing toxicity with the existing toolbox, efforts to broaden the chemotherapeutic window are highly desired. Here, we discover that gossypol (GSP, a natural compound) dramatically enhances the chemosensitivity of cabazitaxel (CTX), even at previously ineffective concentrations. Based on this interesting finding, we exploit a carrier-free chemotherapeutic nano-booster for prostate cancer treatment, which is molecularly co-assembled by GSP and cabazitaxel (CTX). GSP not only readily forms nanoassembly with CTX, but also functions as a chemotherapeutic enhancer that unlocks an ultra-low-dose chemotherapeutic window. Not only that, precise dual-drug nanoassembly confers CTX a significantly larger maximum tolerable dose. As expected, the nano-booster exerts striking therapeutic benefits in mouse prostate tumor xenograft models. This study advances chemotherapeutic window expansion and self-sensitized chemotherapy toward clinical applicability.

Acute lung injury (ALI) has been a kind of acute and severe disease that is mainly characterized by systemic uncontrolled inflammatory response to the production of huge amounts of reactive oxygen species (ROS) in the lung tissue. Given the critical role of ROS in ALI, a Fe₃O₄ loaded bovine serum albumin (BSA) nanocluster (BF) was developed to act as a nanomedicine for the treatment of ALI. Combining with NIR irradiation, it exhibited excellent ROS scavenging capacity. Significantly, it also displayed the excellent antioxidant and anti-inflammatory functions for lipopolysaccharides (LPS) induced macrophages (RAW264.7), and Sprague Dawley rats via lowering intracellular ROS levels, reducing inflammatory factors expression levels, inducing macrophage M2 polarization, inhibiting NF-κB signaling pathway, increasing CD4⁺/CD8⁺ T cell ratios, as well as upregulating HSP70 and CD31 expression levels to reprogram redox homeostasis, reduce systemic inflammation, activate immunoregulation, and accelerate lung tissue repair, finally achieving the synergistic enhancement of ALI immunotherapy. It finally provides an effective therapeutic strategy of BF + NIR for the management of inflammation related diseases.

Objective:To explore the association between healthy lifestyle and co-morbidity of overweight/obesity and myopia in adolescents.Methods:Based on the Shanghai Municipal Dynamic Cohort of Student Common Diseases, we used the group-based trajectory model (GBTM) to establish a model for the development of adolescents' healthy lifestyle trajectories aged 11-14 years and the Cox proportional hazard regression model to assess the effects of different healthy lifestyles on the co-morbidity of overweight/obesity and myopia in adolescents.Results:A total of 2 331 adolescents were included in the analysis. The average follow-up time was 1.87 years, with age of (11.44±0.45) years at the initial follow-up, among 1 106 boys and 1 225 girls. During the 2-year follow-up period, the incidence rates of overweight/obesity, myopia, and co-occurrence of overweight/obesity and myopia were 25.9%, 77.0%, and 13.7%, respectively. GBTM identified 3 healthy lifestyles: the unhealthy group (7.3%), the relatively healthy group (50.2%), and the healthy group (42.5%). The risk of overweight/obesity was lower in the relatively healthy group than in the unhealthy group ( HR=0.694, 95% CI: 0.508-0.947). There was no statistically significant association between the GBTM lifestyle subgroups and myopia ( P>0.05). The risk of co-occurrence of overweight/obesity and myopia was lower in both relatively healthy group and healthy group than in the unhealthy group (relatively healthy group: HR=0.515, 95% CI: 0.348-0.763; healthy group: HR=0.571, 95% CI: 0.384-0.849). Sensitivity analyses showed this result to be stable for myopia, overweight/obesity and myopia co-morbidities. Conclusion:Persistent unhealthy lifestyle increases the risk of overweight/obesity and myopia co-occurrence in adolescents.

Objective:To investigate the effects of insulin-like growth factor 2 (IGF-2) on the expression of postsynaptic density protein 95 (PSD95), synaptophysin-1 (SYN1), and synaptophysin (SYP) in the mouse visual cortex and visual function after monocular deprivation (MD).Methods:Sixty-four SPF male Kunming mice aged 3 weeks were randomly divided into 4 groups: normal control group, MD group, MD+ IGF-2 recombinant protein (MD+ IGF-2) group, and MD+ fluoxetine (FLX) group, with 16 mice in each group.The MD group, MD+ IGF-2 group and MD+ FLX group were treated with right eyelid suturing at the beginning of 3 weeks old and eyelid opening at the end of 5 weeks old.The MD+ IGF-2 group was intraperitoneally injected with IGF-2 recombinant protein during MD.The MD+ FLX group was given fluoxetine via drinking water for 4 weeks after eyelid opening.The normal control group and MD group were injected intraperitoneally with bovine serum albumin every day from 3 to 5 weeks of age.At the end of 5 and 9 weeks of age, subjective visual function was evaluated by fore paw touching ground reflex experiment.At the end of 9 weeks of age, objective visual function was assessed by flash visual evoked potentials.After the mice were sacrificed, the left visual cortex of mice in each group was taken, and the expression of PSD95, SYN1, and SYP was assessed by Western blot.This study was approved by the Ethics Committee of Hunan Children's Hospital (No.HCHDWLL-2022-16). The handling of experimental animals was carried out in accordance with the Guidelines for the Management and Use of Laboratory Animals in Hunan Children's Hospital.Results:At the end of 5 and 9 weeks of age, there were overall significant differences in the success rate of fore paw touching ground among different groups of mice ( F=4.83, 3.36; both P<0.05). At the end of 5 weeks of age, the success rate was lower in MD group and MD+ FLX group than in normal control group, and significantly higher in MD+ IGF-2 group than in MD group, with statistically significant differences (all P<0.05). At the end of 9 weeks of age, the success rate was lower in MD group than in normal control group, and significantly higher in MD+ IGF-2 group and MD+ FLX group than in MD group (all P<0.05). There was a significant overall difference in P2 wave amplitude in F-VEP examination among different groups of mice ( F=13.99, P<0.01). The P2 wave amplitude was significantly lower in MD group than in normal control group and MD+ IGF-2 group (both P<0.01). There was no significant difference in the P2 wave latency of F-VEP among the four groups of mice ( F=2.83, P=0.07). The relative expression levels of PSD95, SYN1 and SYP proteins were 1.00±0.41, 1.00±0.10 and 1.00±0.27 in normal control group, 0.32±0.27, 0.68±0.20 and 0.56±0.28 in MD group, 0.78±0.32, 0.91±0.18 and 0.94±0.22 in MD+ IGF-2 group, 0.89±0.65, 0.98±0.28 and 0.94±0.47 in MD+ FLX group, respectively.There were significant differences in levels of PSD95, SYN1 and SYP in mice visual cortex among different groups ( F=4.24, 5.32, 3.40; all P<0.05). The expressions of PSD95, SYN1 and SYP proteins in the visual cortex were lower in MD group than in normal control group, and higher in MD+ IGF-2 group than in MD group (all P<0.05). Conclusions:Administration of exogenous IGF-2 to mice that underwent MD during the critical period can maintain visual cortex plasticity and protect the visual function to a certain extent.

Objective:To study the impact of fluoride exposure through drinking water on the risk of hypertension among residents in Jishan County, Shanxi Province.Methods:From March to April 2023, a cluster sampling method was used to select permanent residents aged ≥18 years and residing for ≥10 years in 12 villages in drinking water-borne endemic fluorosis areas of Jishan County, Shanxi Province as the survey subjects. A questionnaire survey, physical examination, and morning urinary fluoride level testing were conducted. The least absolute shrinkage and selection operator (Lasso) regression were used to analyze the key influencing factors of hypertension. Restricted cubic spline was used to evaluate the linear relationship between urinary fluoride and hypertension. Logistic regression was used to analyze the impact of urinary fluoride on hypertension.Results:Finally, 2 453 survey subjects were included, aged (62 ± 10) years, including 1 565 patients (63.80%) with hypertension. There were significant differences in the distribution of age, gender, education level, annual household income, body mass index (BMI), and the level and distribution of urinary fluoride between hypertension group and normal blood pressure group ( P < 0.05). The Lasso regression results showed that age, education level, BMI, and urinary fluoride were the key influencing factors of hypertension, with coefficients of 1.04, - 0.12, 0.24 and 0.01, respectively. The results of the restricted cubic spline showed that there was a linear relationship between urinary fluoride and hypertension after adjusting for age, education level, and BMI ( Poverall = 0.018, Pnonlinear = 0.482). The logistic regression results showed that after adjusting for age, education level, and BMI, urinary fluoride > 4.68 mg/L was a risk factor for hypertension ( OR = 1.42, 95% CI: 1.10 - 1.84, P = 0.007). Conclusion:High urinary fluoride is a risk factor for hypertension in drinking water-borne endemic fluorosis areas of Jishan County, Shanxi Province.