Objective:To investigate the effects of Xiaomudan Granules on PERK/eIF2α/ATF4 pathway and lipid metabolism in NAFLD mice.Methods:Totally 60 mice were divided into a normal group of 10 mice and a high-fat diet group of 50 mice. The high-fat diet group was given a high-fat diet to establish a NAFLD mouse model. The high-fat feed group was divided into model group, metformin group, and Xiaomudan Granules low-, medium-, and high-dosage groups according to the random number table method. Xiaomudan Granules low-, medium-, and high-dosage groups were orally administered with concentrated Xiaomudan Granules at dosages of 11.7, 23.4, and 46.8 g/kg, respectively. The metformin group was orally administered with metformin solution at a dose of 0.2 g/kg, while the normal group and model group were orally administered with distilled water of equal volume once a day for 8 consecutive weeks. The liver oil red O staining of mice was observed in each group. The protein expressions of p-PERK, p-eIF2α, ATF4, C/EBP homologous protein (CHOP), CCAAT enhancer binding protein alpha (C/EBPα), C/EBPβ, and steroid regulatory element binding protein 2 (SREBP2) were detected by Western blot. Fluorescence quantitative PCR was used to detect the mRNA expressions of ATF4, CHOP, C/EBPα, C/EBPβ, and SREBP2.Results:The liver tissue structure and steatosis of mice were improved in Xiaomudan Granules groups. Compared with the model group, the expressions of p-PERK/PERK, ATF4, C/EBPα, C/EBPβ, and SREBP2 proteins decreased in Xiaomudan Granules groups and the metformin group ( P<0.01 or P<0.05). The expressions of p-eIF2α/eIF2α and CHOP proteins decreased in the Xiaomudan Granules medium- and high-dosage groups and the metformin group ( P<0.01); compared with the model group, the levels of ATF4, CHOP, C/EBPα, and C/EBPβ mRNA in Xiaomudan Granules groups and the metformin group decreased ( P<0.01), while the levels of SREBP2 mRNA in the Xiaomudan Granules medium- and high-dosage groups and the metformin group decreased ( P<0.01). Conclusion:Xiaomudan Granules may improve liver lipid metabolism and reduce liver fat deposition in NAFLD mice by regulating the PERK/eIF2α/ATF4 pathway.

Objective To investigate the effects of Jiawei Ditan Decoction on neurological function,pentraxin-3(PTX-3),and vascular endothelial growth factor(VEGF)in patients with post-ischemic stroke cognitive impairment(PSCI).Methods A total of 97 patients with PSCI who were admitted to Xuzhou Central Hospital from March 2022 to March 2024 were enrolled and randomly assigned to control group(n=48)or decoction group(n=49)using the envelope drawing method.The control group received conventional treatment,while the decoction group was additionally treated with Jiawei Ditan Decoction.Clinical efficacy,Traditional Chinese Medicine(TCM)syndrome scores,cognitive and functional assessments,laboratory markers,and oxidative stress levels were compared between the two groups.Results The total effective rate in the decoction group was significantly higher than that in the control group(P<0.05).At 3 and 6 months after treatment,TCM syndrome scores in the decoction group were lower than those in the control group(P<0.05).The scores of the Montreal cognitive assessment(MoCA),mini-mental state examination(MMSE),and Barthel index(BI)in the decoction group were higher than those in the control group at 3 months after treatment,while the National Institutes of Health stroke scale(NIHSS)score in the decoction group was lower than that in the control group(P<0.05).At 1 month after treatment,PTX-3 and hypersensitive C-reactive protein(hs-CRP)levels in the decoction group were lower than those in the control group,while VEGF,superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and nitric oxide(NO)levels in the decoction group were higher than those in the control group(P<0.05).Conclusion Jiawei Ditan Decoction exhibits significant effects on improving neurological function and modulating PTX-3 and VEGF levels in patients with PSCI.

Objective To establish a rapid and accurate droplet digital PCR(ddPCR)detection method for detecting Staphylococcus aureus(SA)in laboratory animals and the environment.Methods Using the heat-stable nuclease gene(nuc)of SA as the target gene,a pair of specific primers and probes are designed within its conserved region.Optimize the reaction conditions,test the dynamic range,and evaluate the specificity and stability of the method.Using the same template,test reactions were performed with both ddPCR and real-time quantitative PCR(qPCR)method to assess the interchangeability between the two approaches.Finally,the method is applied to the detection of various clinical samples.Results The kinetic range of the established SA ddPCR method is 100～15 000 copies/μL,with a detection limit of 2.5 copies and a quantification limit of 10 copies;The specificity of this method was tested,and only SA showed positive droplets,while no positive droplets were found for other pathogens;After measuring three parallel samples,the standard deviation and relative standard deviation were calculated.It was found that within the dynamic detection interval of ddPCR,as the target copy number gradually decreased,the relative standard deviation showed an upward trend,but remained below 25％.This result indicates that the detection method has good stability.Conclusions The established ddPCR method for detecting SA has the advantages of high sensitivity,strong specificity,good stability,and good reproducibility.This method can be applied for the detection of SA in laboratory animals.

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Objective To evaluate the impact of lens capsule diameter and capsular tension ring(CTR)fit on the ro-tational stability and visual function of Toric intraocular lenses(IOLs)in patients with high myopia cataract after surgery,and to explore the main influencing factors.Methods A prospective study included 100 eyes of patients with high myopia(axial length≥26 mm)who underwent phacoemulsification cataract extraction combined with Toric IOL implantation at the Cataract Department of Hebei Eye Hospital from September 2023 to March 2024.Based on the ratio of the capsule equivalent diameter measured by anterior segment OCT(CASIA2)to the standard diameter of CTR,the eyes were divided into the matched group(ratio of 0.8-1.2)and the unmatched group.Follow-up was conducted at 1 week,1 month,3 months,6 months,and 12 months postoperatively to observe the rotation angle of Toric IOL,residual astigmatism,uncor-rected visual acuity,and other indicators.Safety was assessed based on intraoperative and postoperative complications.Correlation analysis and multiple linear regression were used to explore the impact of capsule-CTR fit,corneal astigmatism,axial length,and other factors on Toric IOL rotation and postoperative refractive status.Results The mean IOL rotation angle at each postoperative time point in the matched group was significantly smaller than that in the unmatched group,with statistically significant differences between groups at different time points(all P＜0.05).The matched group showed better stability in IOL decentration distance and tilt angle.At 12 months postoperatively,the residual astigmatism in the matched group was(0.53±0.29)D,which was better than that in the unmatched group(0.71±0.34)D(P=0.08).The matched group also had more significant improvement in uncorrected visual acuity and higher subjective visual quality scores.Multiple linear regression analysis showed that capsule-CTR fit and preoperative corneal astigmatism were the main independent factors affecting the rotation angle of Toric IOL(R2=0.52)and residual astigmatism(R2=0.46).Conclu-sion For patients with high myopia cataract,precise capsule-CTR matching based on CASIA2 measurement helps improve the rotational stability of Toric IOLs,reduce residual astigmatism,and enhance postoperative visual quality.Personalized CTR model selection is worth promoting and applying.

Liu Baoyi was a famous febrile disease expert in the late Qing Dynasty,who innovated the theory of latent febrile disease.Liu proposed that the deficiency of kidney essence and the deficiency of healthy qi were the prerequisites for the latent cold e-vil,and the exogenous cold evil was the inducement of the disease.He advocated the treatment of latent febrile disease based on the six meridians,with the two ideas of eliminating evil qi and strengthening healthy qi as the outline,and the general principle of treatment was to relieve heat,nourish yin and expel evil qi.He attached importance to the coordination of medicine and food,made good use of fresh medicine,attached importance to drug processing,emphasized the use of herb processed by herb,and adapted the medicine ac-cording to the syndromes.He preferred to use co-processed products for clinical use.The study of Liu's academic thoughts and medi-cation characteristics in treating latent febrile diseases is of great value in guiding the treatment and medication of various infectious and epidemic diseases with fever characteristics nowadays.

ObjectiveThrough qualitatively and quantitatively analysis of the differences in chemical composition between the combined decoction and single decoction of Huaganjian and comparison of their core efficacy， to explore the rationality of the flexible clinical application of Huaganjian compound preparations and single-flavored dispensing granules. MethodsUltra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS） was used to qualitatively analyze the combined decoction and single decoction samples of Huaganjian， and meanwhile， the contents of four index components（geniposide， paeoniflorin， hesperidin and paeonol） were quantitatively analyzed by high performance liquid chromatography（HPLC）. Nonalcoholic fatty liver disease（NAFLD） rat model induced by high-fat diet was applied to compare the efficacy of combined decoction and single decoction of Huaganjian. A total of 30 male SD rats were randomly divided into the control group， model group， lovastatin group（1.8 mg·kg^-1）， combined decoction group（1.26 g·kg^-1） and single decoction group（1.18 g·kg^-1）. After successful modeling， lovastatin group， combined decoction group and single decoction group were given corresponding doses of drugs by intragastric administration every day， and the control group and model group were given equal amounts of normal saline by intragastric administration， after 4 weeks of administration， the serum and liver tissues were collected， and the contents of alanine aminotransferase（ALT）， aspartate aminotransferase（AST）， total cholesterol（TC）， triglyceride（TG）， low-density lipoprotein cholesterol（LDL-C） and high-density lipoprotein cholesterol（HDL-C） in serum of rats were detected， and the liver pathological examination was carried out by hematoxylin-eosin（HE） staining and oil red O staining， so as to compare differences of their efficacy. ResultsSeventy chemical components were initially identified and attributed from the lyophilized powder of the combined decoction and single decoction samples of Huaganjian， and there was no obvious difference in composition between the two. Further quantitative analysis showed that the contents of geniposide， paeoniflorin， hesperidin and paeonol in the combined decoction samples were significantly increased when compared with those of the single decoction samples（P<0.01）. The pharmacodynamic results showed that compared with the model group， both the combined and single decoction groups of Huaganjian could improve the liver index of NAFLD rats， reduce the serum levels of AST， ALT， TC， TG and LDL-C， increase the serum level of HDL-C， and ameliorate the pathological changes of liver cell steatosis and fat accumulation. However， there was no significant difference in pharmacodynamic effects between the combined decoction group and the single decoction group. ConclusionThere is no significant difference between the combined decoction and single decoction of Huaganjian in terms of chemical composition， but the contents of the four index components show significantly difference. Both of them can significantly improve the fat accumulation and liver function in NAFLD rats. This study provides a reference basis for the rational clinical application and evaluation of famous classical formula compound preparations and single-flavored dispensing granules.

Objective To investigate the inhibitory effect of methionine-γ-lyase(METase) gene MEGL on glioblastoma(GBM) cells and its potential molecular mechanism,and to provide new ideas for the development of MEGL drugs.Methods MEGL expression system mediated by lentivirus was constructed and used to infect human GBM cell lines U87 and snb19,and the cells infected with empty virus were as the control.RT-qPCR and Western blot were used to detect the expression of MEGL gene and protein.The levels of methionine(Met) and its metabolites were detected by liquid chromatography tandem mass spectrometry(LC-MS/MS).The cell proliferation was measured by cell counting method.The changes of cell cycle were detected by flow cytometry,and the expression of cell cycle-related proteins(CDK2,CDK4 and p21) and mitogen-activated protein kinase(MAPK) signaling pathway proteins was determined by Western blot.Results Met levels in U87 and snb19cells stably expressing MEGL gene decreased significantly(t=10.996 and 15.733,respectively,each P <0.001).Furthermore,MEGL significantly suppressed the proliferation of U87(on the 3rd,4th and 5th day,t=14.698,9.746,and 12.263,respectively,each P<0.001) and snb19 cells(on the 3rd,4th and 5th day,t=8.480,6.939,and 13.258,respectively,each P <0.001),inducing a notable increase in the proportion of G0/G1 phase(t=7.997,P P<0.001).MGEL gene decreased the expression of cell cycle related proteins CDK2 and CDK4(t=8.471 and 3.369,P=0.0011 and 0.0281,respectively),while promoting the expression of p21(t=10.761,P <0.001).Moreover,it significantly inhibited the expression of proteins related to the MAPK signaling pathway,including p-p38,p-ERK1/2,and p-JNK(t=3.401,6.493,and9.413,P=0.027 2,0.002 9,and <0.001,respectively).Conclusion The MEGL gene can significantly reduce the intracellular Met levels in U87 and snb19 cells,while also inhibiting cell proliferation and cell cycle progression.This inhibitory effect may be achieved through MAPK signaling pathway.

ObjectiveTo investigate the role and mechanism of Go-Ichi-Ni-San complex subunit 1 （GINS1） in the progression of hepatocellular carcinoma （HCC） and the development of chemotherapy resistance. MethodsThe tumor database GEPIA2 was used to analyze the differential expression of GINS1 between HCC patients and healthy individuals， and pathological tissue samples were collected from 40 HCC patients who were admitted to The Affiliated Tumor Hospital of Xinjiang Medical University and the First Affiliated Hospital of Xinjiang Medical University from May 2017 to January 2021. Immunohistochemical staining was used to measure the difference in the expression of GINS1 between HCC tissue and corresponding adjacent tissue， and the correlation between the expression level of GINS1 and the clinical TNM stage of HCC was analyzed. Western blot was also used to measure the difference in the expression of GINS1 between HCC Huh7/Hep3B/Li-7/MHCC97H cell lines and normal human QSG7701 hepatocytes. The method of lentivirus transfection was used to establish the MHCC97H cell line with stable GINS1 knockdown and its negative control cell line. CCK-8 assay and colony formation assay were used to measure cell proliferative capacity； scratch assay was used to measure cell migration ability； Transwell assay was used to measure cell invasion ability； cells were treated with oxaliplatin to measure their sensitivity to chemotherapy drugs. Nude mice were used to establish a tumor-bearing model and observe the effect of GINS1 knockdown on the growth of HCC in vivo. Western Blot was used to measure the expression levels of the proteins associated with the Notch pathway and the JAK/STAT pathway. The cells were treated with the Notch receptor agonist Jagged-1 to analyze the association between GINS1 and the Notch/JAK/STAT pathway. The independent-samples t test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsThe expression of GINS1 was upregulated in HCC patients， HCC tissue， and HCC cell lines （all P<0.05）， and the expression level of GINS1 was positively correlated with the clinical TNM stage of HCC （r=0.822， P=0.011）. Compared with the negative control cells， the GINS1-knockdown MHCC97H cells showed significant reductions in proliferation， migration， and invasion activities （all P<0.01） and a significantly enhanced sensitivity to oxaliplatin （P<0.01）. Compared with the nude mice in the control group， GINS1 knockdown caused significant inhibition of tumor weight and volume in vivo in nude mice （all P<0.001）. Compared with the negative control cells， the GINS1-knockdown MHCC97H cells showed significant reductions in the expression levels of Notch1， Notch3， p-JAK2， and p-STAT3 （all P<0.05）， while there were no significant differences in the overall expression levels of JAK2 and STAT3 （P>0.05）. After Jagged-1 treatment， the GINS1-knockdown MHCC97H cells showed significant increases in proliferation， migration， and invasion activities and a significant reduction in sensitivity to oxaliplatin， as well as significant increases in the levels of p-JAK2 and p-STAT3 （all P<0.05）. ConclusionGINS1 is upregulated in HCC and can promote HCC progression and chemotherapy resistance through the Notch/JAK2/STAT3 pathway.

Lung cancer is the leading cause of cancer-related deaths worldwide. Radiation pneumonitis is a major complication in lung cancer radiotherapy. Four-dimensional computed tomography (4DCT) imaging provides dynamic ventilation information, which is valuable for lung function assessment and radiation pneumonitis prevention. Many methods have been developed to calculate lung ventilation from 4DCT, but a systematic comparison is lacking. Prediction of radiation pneumonitis using 4DCT-based ventilation is still in an early stage, and no comprehensive review exists. This paper presented the first systematic comparison of functional lung ventilation algorithms based on 4DCT over the past 15 years, highlighting their clinical value and limitations. It then reviewed multimodal approaches combining 4DCT ventilation imaging, dose metrics, and clinical data for radiation pneumonitis prediction. Finally, it summarized current research and future directions of 4DCT in lung cancer radiotherapy, offering insights for clinical practice and further studies.
Humans ; Lung Neoplasms/diagnostic imaging* ; Four-Dimensional Computed Tomography/methods* ; Radiation Pneumonitis/etiology* ; Algorithms ; Lung/radiation effects* ; Pulmonary Ventilation