Background Arsenic is an environmentally harmful substance that causes hepatic insulin resistance and liver damage, increasing the risk of type 2 diabetes mellitus. Objective To explore whether the insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) is involved in insulin resistance in HepG2 cells after arsenic exposure through the peroxisome-proliferator-activated receptor γ (PPAR-γ) / glucose transporter 4 (GLUT4) pathway. Methods Cell viability was determined using cell counting kit 8 (CCK8) and an appropriate NaAsO₂ infection dose was determined. A cellular arsenic exposure model of HepG2 cells was established by four concentrations of NaAsO₂ solution for 24 h (the experiment was divided into four groups: 0, 2, 4, and 8 μmol·L⁻¹); HepG2 cells were firstly treated with pcDNA3.1-IGF2BP2 and pcDNA3.1-NC respectively for 6 h, then with 8 μmol·L⁻¹ NaAsO₂ for 24 h to establish a IGF2BP2 overexpression cell model (the experiment was divided into 4 groups: control, NaAsO₂, NaAsO₂+pcDNA3.1-IGF2BP2, and NaAsO₂+pcDNA3.1-NC); finally the cells were subject to 100 nmol·L⁻¹ insulin stimulation for 30 min. Glycogen and glucose in HepG2 cells were determined by glycogen and glucose assay kits; mRNA expression levels of IGF2BP2 were measured by quantitative real-time PCR; protein expression levels of IGF2BP2, PPAR-γ, and GLUT4 in HepG2 were detected by Western blot (WB); and the binding of IGF2BP2 to PPAR-γ and PPAR-γ to GLUT4 was verified by co-immunoprecipitation (CO-IP) experiment. Results The results of CCK8 experiment showed a dose-effect relationship between NaAsO₂ concentration and cell viability. When the concentration of NaAsO₂ was ≥4 μmol·L⁻¹ , the cell viabilities were lower than that of the control group (P <0.05). With the increasing dose of NaAsO₂ infection, reduced glucose consumption and glycogen levels in HepG2 cells were found in the 2, 4, and 8 μmol·L⁻¹ NaAsO₂ treatment groups compared to the control group (P <0.05). The difference between the mRNA expression level of IGF2BP2 in the HepG2 cells treated with 4 or 8 μmol L⁻¹ NaAsO₂ and the control group was significant (P <0.05). In the IGF2BP2 overexpression cell model, compared with the control group, glucose consumption and glycogen levels were lowered in the NaAsO₂ group (P <0.05), the mRNA expression level of IGF2BP2 and the protein expression levels of IGF2BP2, PPAR-γ, and GLUT4 in the cell membrane were all decreased (P <0.05). Compared with the NaAsO₂ group, the glucose consumption and glycogen levels were increased in the NaAsO₂+pcDNA3.1-IGF2BP2 group (P <0.05), and the mRNA expression level of IGF2BP2 and the protein expression levels of IGF2BP2, PPAR-γ, and GLUT4 in the cell membrane were all increased (P <0.05). The results of CO-IP experiments showed that IGF2BP2 interacted with PPAR-γ as well as PPAR-γ with GLUT4 protein. Conclusion IGF2BP2 is involved in arsenic exposure-induced insulin resistance in HepG2 cells by acting on the PPAR-γ/GLUT4 pathway.

Objective To compare the setup errors between abdominal deep inspiration breath hold (ADIBH) guided by real-time position management (RPM) and free breathing (FB) for breast cancer patients who were treated with intensity modulated radiation therapy (IMRT) after breast-conserving surgery. Methods The data of 60 patients who underwent breast-conserving surgery for left-sided breast cancer and completed IMRT were analyzed retrospectively. Of these patients, 30 received ADIBH technique guided by RPM and 30 received FB technique. Setup errors in translational (X, Y, Z) and rotational (Rx, Ry, Rz) directions were assessed by comparing planning CT and cone-beam CT (CBCT) images for both patient groups. Results Compared with FB group (232 sets of CBCT images), ADIBH (261 sets of CBCT images) significantly reduced setup errors in the translational directions (X, Z) and rotational directions (Rx, Ry, and Rz) (Z values were 3.14, 2.42, 1.45, 1.93, 1.37, respectively; all P<0.05). In the ADIBH group, the difference in setup errors between the patients with BMI <24 kg/m² and those with BMI ≥24 kg/m² was not statistically significantly different (P≥0.05); no significant change in setup errors was detected when comparing the first treatment week with subsequent radiotherapy fractions (P≥0.05). The rotation error in the Ry direction was greater in the first treatment week than subsequent radiotherapy fractions in the FB group (Z=8.02, P=0.02). Conclusion In left-sided breast cancer patients receiving postoperative IMRT, the ADIBH technique demonstrates significantly smaller setup errors compared to FB technique, independent of BMI, thereby improving radiotherapy precision.

Objective:To investigate the effect of oxygen-driven nebulization inhalation combined with invasive mechanical ventilation on sputum viscosity and inflammatory indicators in patients with severe pneumonia complicated by respiratory failure.Methods:A total of 300 patients with severe pneumonia complicated by respiratory failure who received treatment at Lanxi Traditional Chinese Medicine Hospital from June 2021 to June 2023 were included in this prospective study. Using the random number table method, the patients were divided into a control group (conventional invasive mechanical ventilation, n = 150) and a treatment group (conventional invasive mechanical ventilation + oxygen-driven aerosol inhalation, n = 150). The sputum viscosity, inflammatory indicators, and blood gas analysis indicators were compared between the two groups before and after treatment. The occurrence of ventilator-associated pneumonia, mechanical ventilation time, intensive care unit stay duration, and the length of hospital stay were recorded. Results:After treatment, the sputum viscosity grade of the treatment group was significantly different from that of the control group ( Z = 7.09, P < 0.05). The serum levels of interleukin-6 and interleukin-10 in the treatment group were significantly lower, with values of (8.19 ± 0.91) μg/L and (19.26 ± 2.17) μg/L, respectively, compared with (12.01 ± 1.34) μg/L and (32.57 ± 3.85) μg/L in the control group ( t = 28.88, 36.88, both P < 0.05). The arterial partial pressure of oxygen in the treatment group post-treatment was (95.75 ± 3.51) mmHg (1 mmHg = 0.133 kPa), which was significantly higher than (90.14 ± 3.64) mmHg in the control group ( t = 13.58, P < 0.001). The arterial partial pressure of carbon dioxide in the treatment group was (40.65 ± 4.03) mmHg, which was significantly lower than (44.81 ± 4.12) mmHg in the control group ( t = 8.84, P < 0.001). The incidence of ventilator- associated pneumonia in the treatment group was significantly lower than that in the control group ( χ2 = 14.00, P < 0.001). The mechanical ventilation time, intensive care unit stay duration, and the length of hospital stay in the treatment group were significantly shorter compared with the control group ( t = 25.82, 18.23, 15.28, all P < 0.001). Conclusion:Oxygen-driven aerosol inhalation combined with invasive mechanical ventilation can effectively dilute sputum, reduce the patient's inflammatory response, improve blood gas analysis indicators, lower the risk of ventilator- associated pneumonia, and thereby facilitate recovery from severe pneumonia complicated by respiratory failure.

African swine fever(ASF)is an acute and highly pathogenic hemorrhagic disease of pigs,causing huge economic losses to pig industry.In order to quantitatively detect clinical samples of ASF and inactivated ASFV antigens,the IgG of ASF positive serum was used as capture anti-body and the HRP-labeled p72 monoclonal antibody was used as detecting antibody.The standard curve was drawn with the cell-cultured ASFV,and a sandwich ELISA detection of antigen was es-tablished.The specificity,sensitivity and stability of the method were evaluated.The effects of dif-ferent inactivation methods and adjuvant addition on antigen detection were further evaluated.The results showed that the minimum detection limits of the recombinant protein and the ASFV were 0.1 mg/L and 103.7 TCID50/mL,respectively.There was no cross-reaction with five common porcine pathogenic viruses,and the coefficient variations between batches was less than 10％.The total co-incidence rate with real-time fluorescence quantitative PCR was 92％(23/25).The sensitivity of antigen detection was significantly reduced when antigen was treated by BEI inactivation,and the detection results were severely interfered by aluminum adjuvant and nano-adjuvant.In summary,the sandwich ELISA antigen detection method established is specific,sensitive,and repeatable,with a good consistency to the qPCR method,which provides an effective clinical diagnostic meth-od for ASFV antigen.

Objective:To analyze the genes related to platelet activation in essential thrombocythemia (ET)based on transcriptome sequencing technology (RNA-seq ),and to explore the potential targets related to ET thrombosis. Methods:Blood samples from ET patients and healthy individuals were collected for RNA-seq,and differentially expressed lncRNAs,miRNAs,and mRNAs were selected to construct a lncRNA-miRNA-mRNA regulatory network. Differential mRNAs in the regulatory network were enriched and analyzed using Gene Ontology (GO ) and Kyoto Encyclopedia of Genes and Genomes (KEGG).The real-time PCR method was applied to validate differential mRNAs on crucial signaling pathways.Results:A total of 32 lncRNAs (3 up-regulated,29 down-regulated),16 miRNAs (8 up-regulated,8 down-regulated),and 35 mRNAs (27 up-regulated,8 down-regulated)were identified as differentially expressed.Among them,5 lncRNAs,12 miRNAs,and 19 mRNAs constituted the regulatory network.KEGG enrichment analysis showed that the differential mRNAs were related to the platelet activation signaling pathway,and there were 6 differential mRNAs related to platelet activation,namely F2R,ITGA2B,ITGB1,ITGB3,PTGS1,and GP1 BB,which were all up-regulated in their expression.RT-PCR results showed that the expression of five mRNAs including F2R,ITGA2B,ITGB1,ITGB3,and GP1BB were upregulated in ET patients compared with healthy subjects,and consistent with RNA-seq results,while PTGS1 expression was not significantly different.Conclusion:Differential mRNAs in ET patients are related to the platelet activation pathway,and F2R,ITGA2B,ITGB1,ITGB3,and GP1BB mRNAs may serve as novel targets associated with platelet activation in ET.

Objective To explore the dosimetric differences between abdominal deep inspiration breath hold(ADIBH)mode and free breath(FB)mode in intensity modulated radiation therapy(IMRT)for left breast cancer.Methods From July 2022 to May 2023,a total of 22 patients who needed adjuvant radiation therapy after left breast cancer surgery in the hospital were selected as the research objects.The simulated computed tomography(CT)positioning images of ADIBH and FB modes were collected,the planned target volume(PTV)and endangered organs were outlined,the IMRT plan was designed,and the dosimetric param-eters of the two modes were compared.Results There was no significant difference in the mean dose(Dmean),homogeneity index(HI)and conformity index(CI)of PTV between the ADIBH and the FB modes(P＞0.05).Compared with the FB mode,the heart Dmean,V5,V10,V20,V30 and V40 in the ADIBH mode decreased by 2.95 Gy,12.21％,8.26％,6.56％,5.41％and 3.48％,respectively,and the left anterior descending(LAD)coronary artery Dmean,maximum dose(Dmax),minimum dose(Dmin)and V40 decreased by 15.99 Gy,16.10 Gy,0.82 Gy and 13.73％,respectively,with statistical significance(P＜0.05).Compared with the FB mode,the dose and volume of heart irradiation in the ADIBH mode at the same level were significantly reduced.Pearson correlation analysis showed that there was a positive correlation between heart Dmean and LAD Dmean in the ADIBH mode(r=0.72),and between heart Dmean and LAD Dmean in the FB mode(r=0.69).Compared with the FB mode,the left lung Dmean of the ADIBH mode decreased by 0.99 Gy,and the difference was statistically significant(P＜0.05).However,there was no significant difference in left lung V5,right lung Dmean and right breast Dmean between the two breathing modes(P＞0.05).Conclusion ADIBH mode can effectively reduce the dose to the heart and LAD,and play a good protective role.

A biosynthetic gene cluster for the bioactive fungal sesterterpenoids variecolin ( 1) and variecolactone ( 2) was identified in Aspergillus aculeatus ATCC 16872. Heterologous production of 1 and 2 was achieved in Aspergillus oryzae by expressing the sesterterpene synthase VrcA and the cytochrome P450 VrcB. Intriguingly, the replacement of VrcB with homologous P450s from other fungal terpenoid pathways yielded three new variecolin analogues ( 5- 7). Analysis of the compounds' anticancer activity in vitro and in vivo revealed that although 5 and 1 had comparable activities, 5 was associated with significantly reduced toxic side effects in cancer-bearing mice, indicating its potentially broader therapeutic window. Our study describes the first tests of variecolin and its analogues in animals and demonstrates the utility of synthetic biology for creating molecules with improved biological activities.

AIM To investigate the effects of Zhuangyao Shuanglu Tongnao Formula on neuronal apoptosis in rats with cerebral ischemia-reperfusion injury based on the study of oxidative stress and inflammatory response.METHODS The rats were randomly divided into the sham operation group,the model group,the edaravone group(3.0 mg/kg),the low,medium and high dose groups(9.0,18.0,36.0 g/kg)of Zhuangyao Shuanglu Tongnao Formula,with 18 rats in each group.The middle cerebral artery occlusion/reperfusion was conducted by thread embolism method to simulate cerebral ischemia reperfusion injury in rats followed by 6 days corresponding drugs administration.Subsequently,the rats had their neurological function deficit scored by Zeal Longa scoring method;their sizes of cerebral infarction areas measured by TTC staining;their pathological damage and apoptosis of neurons in hippocampal CA1 area of ischemic penumbra of the brain tissue detected by HE staining and TUNEL staining;their SOD activity and levels of GSH,MDA,IL-6,IL-1β,TNF-α in brain tissue detected by kits;and their protein expressions of Bax,Bcl-2,caspase-3,cleaved-capase-3,TLR4,NF-κB p65,Nrf2,HO-1 in rat brain tissue determined by Western blot.RESULTS Compared with the model group,the groups intervened with edaravone,medium and high dose of Zhuangyao Shuanglu Tongnao Formula displayed improvements in the scores of nerve function defects,the rate of cerebral infarction,the rate of neuronal apoptosis,the levels of IL-6,IL-1β,TNF-α and MDA in the ischemic penumbra of brain tissues,the protein expressions of Bax and TLR4,the ratio of cleaved-capase-3/caspase-3 and p-NF-κB p65/NF-κB p65(P＜0.05),the levels of GSH,the activity of SOD and the protein expressions of Bcl-2,Nrf2 and HO-1(P＜0.05).CONCLUSION Being an inhibitor of oxidative stress and inflammatory response,Zhuangyao Shuanglu Tongnao Formula can alleviate brain injury in rats with cerebral ischemia reperfusion injury through the inhibition of neuronal apoptosis and improvement of neural function mediated by the inhibition of TLR4/NF-κB signal pathway and activation of Nrf2/HO-1 signal pathway.

Objective To investigate the protective effect of lovastatin on cardiomyocyte injury induced by hypoxia/reoxygenation and the regulation of protein kinase B(Akt)signal pathway.Methods Rat cardiomyocytes H9c2 were cultured in vitro,and divided into control group,model group,inhibitor group,combined group and experimental-L,-M,-H groups.Control group was cultured with normal medium,and the other 6 groups were treated with hypoxia/reoxygenation.Experimental-L,-M,-H groups were supplemented with 1,2 and 5 μmol·L-1 lovastatin,respectively;inhibitor group was supplemented with 1 μmol·L-1 LY294002;combined group was supplemented with 5 μmol·L-1 lovastatin and 1 pmol·L-1 LY294002.The cell viability was determined by thiazole blue assay,the apoptosis rate was determined by flow cytometry,and the expression level of phosphorylated Akt(p-Akt)was determined by Western blot.Results The cell viabilities of experimental-H,inhibitor,combined,model and control groups were(64.38±7.10)％,(21.64±1.32)％,(51.89±2.25)％,(47.18±6.66)％and(100.00±7.69)％;the cell apoptosis rates were(13.67±1.42)％,(38.52±2.42)％,(21.12±2.27)％,(20.42±3.33)％and(4.93±0.40)％;the relative protein expression levels of p-Akt were 0.54±0.04,0.04±0.01,0.25±0.03,0.20±0.01 and 0.45±0.02,respectively.The differences of above indicators were statistically significant between the experimental-H group and the model group,as well as between the combined group and the experimental-H and inhibitor groups(all P＜0.05).Conclusion Lovastatin can protect against hypoxia/reoxygenation cardiomyocyte injury by activating the Akt signaling pathway.

Perimenopausal syndrome （MPS）， a common endocrine system disease， is one of the diseases responding specifically to traditional Chinese medicine （TCM）. The China Association of Chinese Medicine organized experts in endocrinology， gynecology， and interdisciplinary fields of both Western and Chinese medicine to discuss the advantages and challenges of diagnosing and treating MPS with Western medicine， TCM， and integrative medicine. Experts at the conference believe that MPS is initiated by estrogen decline and rooted in deficiency， with the pathogenesis being imbalance between Yin and Yang in the kidney. The hormone replacement therapy in Western medicine for menopause can rapidly alleviate related symptoms by quickly restoring the estrogen level and timely detect and delay complications of menopause， whereas such a therapy has certain risks， necessitating close monitoring of adverse reactions. Moreover， the various contraindications and precautions limit the clinical application of the hormone replacement therapy. TCM has advantages in synergistically alleviating symptoms such as hot flashes， sweating， sleep disorders， and emotional abnormalities of MPS without causing obvious adverse reactions. However， its efficacy is slower than the hormone replacement therapy， and the TCM evidence for preventing and treating complications of menopause remains unclear. Three suggestions were proposed for the future development of both Western and TCM for ameliorating MPS. First， an integrated diagnosis and treatment system for MPS with both Western and Chinese medicine should be established. Second， high-quality evidence-based interventions for MPS should be developed with TCM alone or in combination with Western medicine. Third， efforts should be made to promote the new TCM drug development and the interdisciplinary cooperation for treating MPS.