[摘 要] 目的：探讨高良姜素（Gal）调控AMPK/mTOR/ULK1信号通路对胃癌细胞凋亡和自噬的影响及其机制。方法：将胃癌NCI-N87细胞分为对照组、多索吗啡（DM）组、Gal低剂量（Gal-L）组、Gal高剂量（Gal-H）组、Gal-H + DM组。采用MTT法、流式细胞术、划痕愈合实验和Transwell实验分别检测各组细胞的增殖、凋亡、迁移和侵袭能力，WB法检测PCNA、C-caspase-3、免疫逃逸相关蛋白（B7H1）、EMT和AMPK/mTOR/ULK1信号通路蛋白的表达水平。建立裸鼠NCI-N87细胞移植瘤模型，观察Gal和5-FU对移植瘤的抑制效果。结果：与对照组比较，DM组NCI-N87细胞增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著升高（均P < 0.05），细胞凋亡率、C-caspase-3、E-cadherin、LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著降低（均P < 0.05）；Gal-L组和Gal-H组NCI-N87细胞的增殖活性、划痕愈合率和侵袭细胞数、N-cadherin、vimentin、PCNA、B7H1、p62和p-mTOR/mTOR蛋白表达均显著降低（均P < 0.05），细胞凋亡率、C-caspase-3、E-cadherin、LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK和p-ULK1/ULK1蛋白表达均显著升高（均P < 0.05）；DM可部分逆转Gal对NCI-N87细胞恶性生物学行为的抑制作用（P < 0.05）；与对照组比较，Gal组和5-FU组裸鼠移植瘤体积和质量均显著降低，肿瘤组织细胞凋亡率显著升高（P < 0.05）。结论：Gal可促进胃癌NCI-N87细胞自噬和凋亡，抑制其增殖、迁移和侵袭，可能与激活AMPK/mTOR/ULK1信号通路有关。

ObjectiveThis paper aims to analyze the damage degree of muscle tone in rats with spasticity of cerebral apoplexy （SCA） and the expression of Nestin and β-catenin in the M1 region of the cerebral cortex， thereby investigating the action mechanism of different doses of Shaoyao Gancaotang on rats with SCA. MethodsThe rats were randomly divided into a blank group， a model group， a positive control group （baclofen， 5.25 mg·kg^-1）， and low， medium， and high-dose groups of Shaoyao Gancaotang （2.1， 4.2， 8.4 g·kg^-1）， with nine rats in each group. A rat model with SCA was established by using a modified phrenic nerve block combined with intraventricular injection of anhydrous ethanol. Following behavioral scoring to confirm model validity， drug interventions were conducted. Neurological deficits and muscle tone were evaluated by behavioral assessments. The open field test was used to measure locomotor distance. Transmission electron microscopy was employed to examine the synaptic structures. Skeletal muscle adenosine triphosphate （ATP）ase staining was used to analyze myofibrillar changes. Hematoxylin and eosin （HE） staining was used to observe the histomorphological changes. Immunohistochemistry， Real-time polymerase chain reaction （Real-time PCR）， and Western blot were employed to detect mRNA levels and protein expressions of Nestin and β-catenin in the M1 region of the cerebral cortex. ResultsCompared with the blank group， rats in the model group exhibited significantly increased neurological deficit scores （P<0.01）， markedly elevated muscle tone scores （P<0.01）， substantially reduced locomotor distance （P<0.01）， prominent structural swelling and blurring， severe destruction of cerebral cortical cells， a significant increase in the proportion of skeletal muscle ATPase type Ⅰ fibers （P<0.01）， a significant decrease in mRNA levels and protein expression of Nestin （P<0.01）， and a significant increase in mRNA levels and protein expression of β-catenin （P<0.01）. Compared with the model group， the Shaoyao Gancaotang group exhibited reduced neurological deficit scores and muscle tone scores in rats with SCA （P<0.01） and increased locomotor distance （P<0.01）. Transmission electron microscopy revealed clearer and more intact synaptic structures in the rats from the Shaoyao Gancaotang group， with increased vesicle numbers and improved morphology. HE staining revealed intact neuronal cell structures with regular arrangement and reduced vacuolated cells in the rats from Shaoyao Gancaotang. ATPase staining result indicated a decreased proportion of type Ⅰ muscle fibers in the rats from the Shaoyao Gancaotang group （P<0.01）. Real-time PCR results demonstrated increased mRNA expressions of Nestin and β-catenin in the rats from the Shaoyao Gancaotang group （P<0.01）. Immunohistochemistry and western blot analyses indicated elevated protein expressions of Nestin and β-catenin in rats with SCA from the Shaoyao Gancaotang group （P<0.05， P<0.01）. ConclusionShaoyao Gancaotang may improve neurological function impairment and limb spasticity in model rats with SCA by regulating the proliferation and differentiation of neural stem cells in the cerebral cortex M1 region.

Objective To analyze the newly reported HIV-1 infection in several prefectures of Hubei Province，and analyze its influencing factors. Methods The limiting antigen avidity enzyme immunoassay(LAg-avidity EIA,LAg) was conducted on HIV-1 positive samples confirmed by Western blot of Hubei in 2017-2022. The demographic characteristics of the newly infected samples were analyzed by χ2 test.Logistic regression model was used to analyze influencing factors of new infection rate and predict the factors associated with the HIV-1 recent infection. Results There were 403 new cases of HIV-1 from 2017 to 2022 in several prefectures of Hubei Province, of which 77 were newly infected sorted by LAg,with a new infection rate of 19.11%. The newly confirmed HIV-1 persons of whom aged ≤24 years (40.00% new infection ratio), unmarried (29.41%), college or above (31.37%), and from Voluntary counseling and testing testing(VCT) clinics (40.00%) had a higher proportion of new infections, and the difference was statistically significant. Multivariate Logistic regression analysis showed that age ≤24 years old (aOR=4.346,95%CI: 1.342-14.075) and screening from the VCT clinic (aOR=6.761,95%CI: 1.460-31.319) were more likely to be newly infected. Conclusion The proportion of new HIV infection in several prefectures of Hubei province is relatively low in recent years.Further effective publicity and intervention measures for young students and the construction of VCT clinic should be continuously promoted to achieve early diagnosis and treatment.

ObjectiveTo explore the possible mechanisms of Modified Shoutai Pill （寿胎丸加味方， MSP） in treating polycystic ovary syndrome （PCOS） with hyperandrogenism， insulin resistance， and miscarriage， focusing on inflammatory response and endometrial receptivity. MethodsThirty female SPF-grade SD rats with regular estrous cycles and in proestrus， and 15 male SPF-grade SD rats were housed together in a 2∶1 ratio at 18：00. At 8：00 next morning， rats showing abundant sperm and vaginal plugs were considered pregnant on the day 0.5. The 30 pregnant rats were randomly divided into three groups， normal group， model group， and MSP group， with 10 rats in each group. From day 0.5 to day 13.5 of pregnancy， the MSP group was given 26.6 g/（kg·d） of the MSP via gavage twice a day for 14 consecutive days. The normal group and the model group received 4 ml of normal saline daily. From day 7.5 to day 13.5 of pregnancy， the rats in the model group and MSP group were intraperitoneally injected with dihydrotestosterone （DHT） and insulin （INS） for 7 consecutive days to establish a PCOS model with hyperandrogenism， insulin resistance， and miscarriage. On day 13.5 of pregnancy， an oral glucose tolerance test （OGTT） was performed to measure blood glucose levels at 0， 30， 60， 90， and 120 minutes. On day 14.5， serum level of progesterone （P₄）， estradiol （E₂）， fasting insulin （FINS）， interleukin-6 （IL-6）， and tumor necrosis factor-alpha （TNF-α） were measured by ELISA. The insulin resistance index （HOMA-IR） was calculated. Embryo implantation， miscarriage rate， and average number of live fetuses were observed. Uterine tissue pathology was examined by HE staining， and mRNA expression of Il-6， Tnf-α， leukemia inhibitory factor （Lif）， homeobox gene 10 （Hoxa10）， prolactin family 8 subfamily A member 2 （Prl8a2）， and insulin-like growth factor-binding protein 1 （Igfbp1） in the uterine tissue was detected by qRT-PCR. ResultsCompared with the normal group， the model group had significantly higher blood glucose level at 0， 30， 60， 90， and 120 minutes， increased miscarriage rate， elevated HOMA-IR， decreased average number of live fetuses， lower level of P₄ and E₂， higher level of IL-6， TNF-α， and FINS， and higher mRNA expression of Il-6 and Tnf-α in the uterine tissue. The mRNA expression of Lif， Hoxa10， and Prl8a2 was reduced （P＜0.05 or P＜0.01）. The uterus had a dark red color， visible areas of bleeding， fewer embryos with developmental abnormalities， and increased placental necrosis. Pathological examination revealed thrombus in the decidual layer， unclear decidual cell morphology， loose arrangement， scattered distribution， edema degeneration in the cytoplasm， and nuclear shrinkage or disappearance， with extensive infiltration of inflammatory cells. In contrast， compared with the model group， the MSP group showed significantly lower blood glucose level at 0， 30， 60， 90， and 120 min， reduced miscarriage rate， lower HOMA-IR， increased number of live fetuses， higher level of P₄ and E₂， and lower level of IL-6， TNF-α， and FINS. The mRNA expression of Il-6 and Tnf-α in the uterine tissue was lower， while the expression of Lif， Hoxa10， and Prl8a2 mRNA was higher （P＜0.05 or P＜0.01）. There was significant improvement in uterine and embryo conditions， as well as in uterine tissue pathology. ConclusionThe MSP can reduce the miscarriage rate in a PCOS model with hyperandrogenism， insulin resistance， and miscarriage. Its mechanism may involve inhibiting inflammation， improving endometrial receptivity， and restoring the defects in endometrial decidualization.

Objective : To investigate the antidepressant effects and the underlying mechanisms of tetrahydrocurcumin(THC) and its nanoparticle formulation(THCN). Methods : Forty-six male ICR mice were randomly divided into Con group, LPS group, THC group, THCN group and SER group. A mouse depression model was established by intraperitoneal administration of LPS. The anxiety and depression-like behaviors of mice were evaluated by open field test(OFT) and forced swimming test(FST). Myelin staining was applied to assess the extent of demyelination in the prefrontal cortex of the mice. The prefrontal cortex and hippocampus were further examined for the expression levels of glial fibrillary acidic protein(GFAP) and Toll-like receptor 4(TLR4) through quantitative immunofluorescence assays. Results : Compared with the Con group, the LPS group showed increased anxiety-like and depressive-like behaviors in both the long-term and short-term experiments(P<0.05); the degree of demyelination increased in the LPS group of the long-term experiment(P<0.01); the expression of GFAP was reduced in the LPS group of the short-term experiment(P<0.01), while the expression of TLR4 increased(P<0.05); the expression of TLR4 decreased in the THC group(P<0.01); the expression of GFAP in the prefrontal cortex of the THCN group was reduced(P<0.01), while the expression of TLR4 increased(P<0.05). Compared with the LPS group, the THC group showed reduced depressive-like behaviors in the long-term experiment(P<0.05), while the anxiety-like and depressive-like behaviors of the THCN group and the SER group were reduced(P<0.05), and the anxiety-like and depressive-like behaviors of the THC group and the THCN group were reduced in the short-term experiment(P<0.05); the degree of demyelination was reduced in the THC group, THCN group and SER group in the long-term experiment(P<0.05); the expression of GFAP increased in the THC group of the short-term experiment(P<0.05), while the expression of TLR4 was reduced(P<0.05), and the expression of GFAP increased in the THCN group(P<0.05). Compared with the THC group, the THCN group and the SER group showed reduced anxiety-like behaviors in the long-term experiment(P<0.05); the expression of GFAP in the prefrontal cortex of the THCN group was reduced in the short-term experiment(P<0.05), while the expression of TLR4 in the hippocampal DG area increased in the short-term experiment(P<0.01). Conclusion Tetrahydrocurcumin and its nanoparticle formulation both exert significant ameliorative effects on depression-like behaviors and demyelination in mice induced by lipopolysaccharide. The antidepressant mechanism of THC appears to be mediated through the down-regulation of TLR4 and the up-regulation of GFAP. The mechanism underlying the antidepressant action of THCN seems predominantly focused on the enhancement of GFAP expression.

OBJECTIVE To investigate the improvement effect and potential mechanism of Qingxue xiaozhi jiangtang formula on insulin resistance （IR） in type 2 diabetes mellitus （T2DM） rats. METHODS T2DM rat model was established by intraperitoneal injection of 30 mg/kg streptozotocin combined with high-fat and high-sugar diet. The rats were randomly divided into normal control group， model group， Qingxue xiaozhi jiangtang formula low-dose and high-dose groups （6.525， 13.05 g/kg， calculated by raw material） and metformin group （positive control， 0.18 g/kg）， with 8 rats in each group. Administration groups were given relevant medicine intragastrically； normal control group and model group were given constant volume of normal saline intragastrically， once a day， for consecutive 6 weeks. Body mass and fasting blood glucose （FBG） were determined， and oral glucose tolerance test was conducted. Serum fasting insulin （FINS） level was measured to calculate the insulin resistance index （HOMA-IR） and insulin sensitivity index （ISI）. Additionally， the level of serum lipids， liver function， oxidative stress indicators and inflammatory factors were assessed. The phosphorylation levels of kinase R-like endoplasmic reticulum kinase （PERK） and forkhead box O1 （FOXO1） protein in liver tissue of rats were determined. RESULTS Compared with model group， the body weight， ISI， the levels of high-density lipoprotein cholesterol and superoxide dismutase were increased significantly in Qingxue xiaozhi jiangtang formula high-dose group and metformin group （P＜0.05）； FBG， blood glucose level at 120 minutes of glucose loading， area under the curve of glucose， FINS， HOMA-IR， low-density lipoprotein cholesterol， total cholesterol， triglyceride， alanine transaminase， aspartate transaminase， alkaline phosphatase， malondialdehyde， interleukin-6， tumor necrosis factor-α， and C-reactive protein levels were significantly reduced （P＜ Δ0.05）； the pathological damage of liver tissue had significantlyimproved， and the phosphorylation levels of PERK and FOXO1 proteins in liver tissue were significantly decreased （P＜0.05）. CONCLUSIONS Qingxue xiaozhi jiangtang formula can regulate glucose and lipid metabolism， inflammation factor and oxidative stress levels， and alleviate insulin resistance in T2DM rats. Its mechanism of action may be related to the inhibition of the PERK/FOXO1 signaling pathway.

Objective To explore the accuracy of artificial intelligence-based diagnosis of ovarian malignant tumors and the identification of benign and malignant tumors under transabdominal scanning and transvaginal scanning methods. Methods A dataset of transabdominal and transvaginal two-dimensional ultrasound images was used and the images were preprocessed to enhance quality. The region of interest was segmented and divided into a training set and a test set. A convolutional neural network (CNN) was trained on the images in the training set, and the accuracy of the model on the test set was calculated. Results Transvaginal scanning was 14% more accurate in diagnosing malignant ovarian tumors than transabdo-minal scanning on the test set. For identifying the benign and malignant ovarian tumors containing cystic components, a mixture of transvaginal and transabdominal scanning increased the accuracy by 9.7% over transabdominal scanning alone. Conclusion CNN can identify ovarian malignant tumors under both scanning methods, but the accuracy of transvaginal scanning is higher than that of transabdominal scanning, and the CNN model has a higher accuracy in identifying benign and malignant ovarian tumors under transvaginal scanning.

Objective To develop and validate a model to predict the risk of radiation-induced lung injury (RILI) and assess its clinical feasibility. Methods Clinical data from 125 patients with non-small cell lung cancer (NSCLC) were included in the study. The patients were divided into training group (88 cases) and validation group (38 cases). Key predictive factors were identified using univariate and multivariate logistic regression analyses combined with least absolute shrinkage and selection operator (LASSO) regression. A predictive model was constructed and evaluated using a nomogram, receiver operating characteristic (ROC) curve, calibration curve, and decision curve analysis. Results The key variables identified by the model were tumor volume (P = 0.017), Eastern Cooperative Oncology Group performance status score (P = 0.035), 95% of the minimum dose to the target volume (P = 0.028), percentage of bilateral lung volume receiving 20 Gy of radiation (P < 0.001), and neutrophil-to-lymphocyte ratio (P = 0.021). The ROC curve showed that the areas under the curve (AUC) for the model in the training and validation groups were 0.987 and 0.992, respectively, indicating good predictive ability. The calibration curve and decision curve further confirmed the accuracy and clinical practicability of the model. Conclusion The predictive model proposed in this study can accurately assess the risk of developing RILI in patients with NSCLC who have undergone radiotherapy, demonstrating its potential value in clinical practice.

In view of the few studies on the influence of Armillaria spp. infection on the content of the chemical components in different parts of Polyporus umbellatus sclerotia, this study determined the biomass of P. umbellatus sclerotia and the contents of ergosterol, polyporusterone A, polyporusterone B and polysaccharide in the separated cavity wall of the sclerotia and the uninfected part of the sclerotia in different harvesting years under the conditions of A. gallica and A. mellea infection respectively. According to the difference of content and dynamic changes of the polysaccharide and the steroid substances, the superior Armillaria sp. was screened to obtain the best harvest years of P. umbellatus. Using HPLC and UV-VIS spectrophotometry methods, the contents of ergosterol, polyporusterone A, polyporusterone B and polysaccharide in P. umbellatus sclerotia infected by the two Armillaria spp. in different years were determined. In addition, the differentially expressed genes related to P. umbellatus polysaccharide synthesis were screened according to the transcriptomic data of different parts of P. umbellatus after A. mellea infection. With the increase of years, the biomass of sclerotia infected by different Armillaria spp. had significant differences, and there were significant differences in the four components of sclerotia. The four components of the separated cavity wall of the sclerotia were significantly higher than those of the uninfected part. The best harvest time was the third year after cultivation. Transcriptomic analysis showed that the infection of Armillaria spp. could significantly promote polysaccharide synthesis, which provided a basis for polysaccharide content determination at the molecular level. The study clarified the influence of different Armillaria spp. infection on the accumulation of chemical components of P. umbellatus sclerotia, laying a foundation for exploring the symbiosis mechanism and provided a scientific clue for screening superior Armillaria sp. and guiding the artificial cultivation of P. umbellatus sclerotia.

The chemical constituents in dried roots of Atractylodes macrocephala were investigated in this study. Through utilizing normal-phase silica gel and ODS column chromatography, TLC, and semi-preparative HPLC, 4 new sesquiterpenoids were purified from the ethyl acetate extract of A. macrocephala. By various spectroscopic techniques, such as 1D and 2D NMR, HR-ESI-MS, IR, UV, and CD, their structures were identified as atractylmacron A (1), 9β-hydroxyasterolide (2), atractylenolide H (3), atractylenolide J (4). Compound 1 possesses a rare 6/7 bicyclic skeleton.