OBJECTIVE To investigate the improvement effects and mechanism of astragaloside Ⅳ （AS- Ⅳ ） on lipopolysaccharide （LPS）-induced neuroinflammation. METHODS BV2 cells were divided into control group， LPS group， AS-Ⅳ groups at concentrations of 20 and 40 μmol/L， and dexamethasone group （2 μmol/L）. Except for control group， neuroinflammation model was established with LPS （1 μg/mL） in other groups after medication. The levels of inflammatory factors [interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， and nitric oxide （NO）] in cell supernatant were measured in each group. Mice were randomly divided into normal group， model group， positive control group （Aspirin enteric-coated tablet， 20 mg/kg）， AS-Ⅳ low- and high-dose groups （10， 20 mg/kg）， with 6 mice in each group. Mice in each group were administered the corresponding drug/normal saline via gavage/intraperitoneal injection， once a day， for 14 consecutive days. Except for normal group， other groups were intraperitoneally injected with LPS （250 μg/kg） 1 hour after daily administration of the drug/normal saline to establish neuroinflammation model. Serum levels of IL-6 and TNF-α were measured 2 h after the last medication； histopathological morphology of cerebral tissue in mice were observed； the co-localization of inducible nitric oxide synthase （iNOS）/ionized calcium binding adapter molecule 1 （Iba1） and CD206/Iba1 in the cerebral cortex region of mice was observed； the expressions of proteins related to the nuclear factor-κB （NF-κB）/mitogen-activated protein kinase （MAPK） signaling pathway in brain tissue of mice were also determined， including NF-κB p65， phosphorylated NF-κB p65（p-NF-κB p65）， p38 MAPK， phosphorylated p38 MAPK （p-p38 MAPK）， extracellular signal-regulated kinase （ERK）， and phosphorylated ERK （p-ERK）. RESULTS In the cell experiments， compared with control group， the levels of IL-6， TNF- α and NO in the cell supernatant of the LPS group were increased significantly （P＜0.05）； compared with LPS group， the levels of IL-6， TNF-α and NO were decreased significantly in the administration groups （P＜0.05）. In the animal experiments， compared with the normal group， the serum levels of IL-6 and TNF- α， the number of iNOS/Iba1 co-localization positive cells in the cerebral cortex， and the phosphorylation levels of p38 MAPK， NF- κB p65 and ERK proteins in brain tissue were all significantly increased/elevated in model group （P＜0.05）； the number of CD206/ Iba1 co-localization positive cells in the cerebral cortex region significantly decreased （P＜0.05）. The neurons in the cerebral cortex and the CA3 region of the hippocampus displayed a disordered arrangement. Compared with model group， above quantitative indexes of mice were all reversed significantly in administration groups （P＜0.05）； the neuronal cells in the cerebral cortex and the CA3 region of the hippocampus exhibited a relatively orderly arrangement. CONCLUSIONS AS-Ⅳ may inhibit the activation of the NF-κB/MAPK signaling pathway， promote the M2-type polarization of microglia， and thereby suppress neuroinflammatory responses.

Based on the theory of 'spleen governing the containment of blood' in The Classic of Difficult Issues （《难经》）， it is proposed that medication toxicity damaging the middle jiao （焦） and failure of the spleen to contain blood constitute the core pathogenesis of cancer therapy induced thrombocytopenia （CTIT）. It is believed that the anti-tumor drug toxicity directly hits middle jiao， leading to spleen qi depletion， impairment of transportation and transformation， and insufficiency of the source of blood production. Consequently， the vessels and collaterals are deprived of nourishment and become structurally damaged. Blood fails to be contained and stored within the vessels， resulting in extravasation beyond the vascular system. Persistent retention of drug toxicity may further cause stasis due to blood deviating from its normal pathway and gradually consume the essence. In view of this root deficiency with branch excess， it is proposed that the treatment should focus on fortifying spleen， boosting qi and containing blood， with the aim of restoring spleen and stomach transportation functions， repairing vascular damage， and recovering the spleen's blood-containing function. According to individual syndrome patterns， prescriptions may be supplemented with herbs that stabilize vessels and consolidate collaterals， remove stasis and promote regeneration， and replenish essence and nourish marrow， providing new perspectives for the prevention and treatment of CTIT with traditional Chinese medicine.

Objective To observe the value of ultrashort echo time(UTE)-MRI for evaluating pulmonary nodules.Methods Totally 58 patients with pulmonary nodules detected with CT were prospectively enrolled,and UTE-MRI was performed.Taken CT as the referent standard,the value of UTE-MRI for evaluating the diameter,composition,lung imaging reporting and data system(Lung-RADS)type and radiographic signs of pulmonary nodules was analyzed.Results CT detected 66 pulmonary nodules with a diameter of(11.60±5.20)mm in 58 patients,including 29 solid nodules,24 partially solid nodules and 13 ground-glass nodules.There were 14 Lung-RADS type 2,12 type 3,12 type 4A,11 type 4B and 17 type 4X lung nodules,among which 12 were found with lobulated sign,and 14 were found with spiculated sign.UTE-MRI detected 63 nodules with a diameter of(11.34±4.82)mm,including 25 solid nodules,28 partially solid nodules and 10 ground-glass nodules.There were 12 Lung-RADS type 2,12 type 3,12 type 4A,11 type 4B and 16 type 4X lung nodules.Among which 10 nodules were found with lobulated sign and 11 were found with spiculated sign.No significant difference of the measured diameters of lung nodules was found between UTE-MRI and CT(P=0.803),and the results of UTE-MRI and CT had good correlation and consistency(rs=0.953,ICC=0.946),which also had good consistency for assessing nodule composition and Lung-RADS type(Kappa=0.871,0.960).Meanwhile,no significant difference of the display rates of lobulated nor spiculated signs was noticed between UTE-MRI and CT(both P＞0.05).Conclusion UTE-MRI was helpful for evaluating pulmonary nodules,with efficacy comparable to CT.

Objective To establish an efficient and stable model of pelvic inflammatory disease in rats via a non-surgical method,and to evaluate its application in pharmacodynamic testing.Methods Female Sprague-Dawley rats were divided randomly into the following groups:control group;model group with phenol for 7 d;model group with phenol for 10 d;treatment group modeled with phenol;model group with low concentration of bacteria;model group with high concentration of bacteria;and treatment group modeled with bacteria.Rats in the model and treatment groups were injected with 25%phenol gel and 2×107 or 2×108 Escherichia coli and Staphylococcus aureus mixture via a non-surgical method,to construct a rat model of pelvic inflammatory disease.Rats in the treatment groups received the Chinese patent medicine Jingangteng capsules by gavage,and rats in the control group received the same volume of solvent solution.The health status,weight changes,and uterine appearance were monitored and the uterine coefficient was calculated.Pathological changes in the uterus and fallopian tubes,endometrial thickness,and number of glands were detected by hematoxylin and eosin staining.Serum levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor(TNF)-α were detected by enzyme-linked immunosorbent assay.Protein expression of the macrophage marker CD68 was detected by immunofluorescence.Expression of Toll-like receptor 4(TLR4)/nuclear factor(NF)-κB pathway-related proteins in the uterus was detected by Western blot.Results The mortality rate in the model group was only 5%.Compared with the control group,model rats showed decreased body weight,increased uterine coefficient,pathological changes in the uterus and Fallopian tubes,thinner endometrium,fewer glands,significantly higher serum levels of IL-1β,IL-6,and TNF-α and more macrophages in the uterine tissue,and activation of the TLR4/NF-κB signaling pathway.The 7 d phenol and low-concentration bacterial solution models were judged to be mild pelvic inflammatory disease models,and the 10 d phenol and high-concentration bacterial solution models were considered severe pelvic inflammatory disease models.Treatment with Jingangteng capsules relieved the pathological symptoms in the uterus and fallopian tubes,in line with the efficacy evaluation of clinical pelvic inflammatory disease.Conclusions We established rat models of pelvic inflammatory disease using phenol and a mixed bacterial solution via a non-surgical method,to simulate the different pathological states of pelvic inflammatory disease caused by different factors.These models will be suitable for evaluating drug efficacy and elucidating the pathological mechanism of pelvic inflammatory disease.

Objective To observe the value of ultrashort echo time(UTE)-MRI for evaluating pulmonary nodules.Methods Totally 58 patients with pulmonary nodules detected with CT were prospectively enrolled,and UTE-MRI was performed.Taken CT as the referent standard,the value of UTE-MRI for evaluating the diameter,composition,lung imaging reporting and data system(Lung-RADS)type and radiographic signs of pulmonary nodules was analyzed.Results CT detected 66 pulmonary nodules with a diameter of(11.60±5.20)mm in 58 patients,including 29 solid nodules,24 partially solid nodules and 13 ground-glass nodules.There were 14 Lung-RADS type 2,12 type 3,12 type 4A,11 type 4B and 17 type 4X lung nodules,among which 12 were found with lobulated sign,and 14 were found with spiculated sign.UTE-MRI detected 63 nodules with a diameter of(11.34±4.82)mm,including 25 solid nodules,28 partially solid nodules and 10 ground-glass nodules.There were 12 Lung-RADS type 2,12 type 3,12 type 4A,11 type 4B and 16 type 4X lung nodules.Among which 10 nodules were found with lobulated sign and 11 were found with spiculated sign.No significant difference of the measured diameters of lung nodules was found between UTE-MRI and CT(P=0.803),and the results of UTE-MRI and CT had good correlation and consistency(rs=0.953,ICC=0.946),which also had good consistency for assessing nodule composition and Lung-RADS type(Kappa=0.871,0.960).Meanwhile,no significant difference of the display rates of lobulated nor spiculated signs was noticed between UTE-MRI and CT(both P＞0.05).Conclusion UTE-MRI was helpful for evaluating pulmonary nodules,with efficacy comparable to CT.

Objective To establish an efficient and stable model of pelvic inflammatory disease in rats via a non-surgical method,and to evaluate its application in pharmacodynamic testing.Methods Female Sprague-Dawley rats were divided randomly into the following groups:control group;model group with phenol for 7 d;model group with phenol for 10 d;treatment group modeled with phenol;model group with low concentration of bacteria;model group with high concentration of bacteria;and treatment group modeled with bacteria.Rats in the model and treatment groups were injected with 25%phenol gel and 2×107 or 2×108 Escherichia coli and Staphylococcus aureus mixture via a non-surgical method,to construct a rat model of pelvic inflammatory disease.Rats in the treatment groups received the Chinese patent medicine Jingangteng capsules by gavage,and rats in the control group received the same volume of solvent solution.The health status,weight changes,and uterine appearance were monitored and the uterine coefficient was calculated.Pathological changes in the uterus and fallopian tubes,endometrial thickness,and number of glands were detected by hematoxylin and eosin staining.Serum levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor(TNF)-α were detected by enzyme-linked immunosorbent assay.Protein expression of the macrophage marker CD68 was detected by immunofluorescence.Expression of Toll-like receptor 4(TLR4)/nuclear factor(NF)-κB pathway-related proteins in the uterus was detected by Western blot.Results The mortality rate in the model group was only 5%.Compared with the control group,model rats showed decreased body weight,increased uterine coefficient,pathological changes in the uterus and Fallopian tubes,thinner endometrium,fewer glands,significantly higher serum levels of IL-1β,IL-6,and TNF-α and more macrophages in the uterine tissue,and activation of the TLR4/NF-κB signaling pathway.The 7 d phenol and low-concentration bacterial solution models were judged to be mild pelvic inflammatory disease models,and the 10 d phenol and high-concentration bacterial solution models were considered severe pelvic inflammatory disease models.Treatment with Jingangteng capsules relieved the pathological symptoms in the uterus and fallopian tubes,in line with the efficacy evaluation of clinical pelvic inflammatory disease.Conclusions We established rat models of pelvic inflammatory disease using phenol and a mixed bacterial solution via a non-surgical method,to simulate the different pathological states of pelvic inflammatory disease caused by different factors.These models will be suitable for evaluating drug efficacy and elucidating the pathological mechanism of pelvic inflammatory disease.

Objective:To explore the molecular mechanism by which the methionine adenosyltransferase 2A (MAT2A)/β-catenin/zinc finger E-box binding homeobox 1 (ZEB1)/epithelial-mesenchymal transition (EMT) pathway regulates neural tube defect (NTD) through intracellular S-adenosylmethionine (SAM).Methods:A mouse NTD model was induced using the SAM metabolic disorder inhibitor ethionine. Eighty specific pathogen-free C57BL/6 mice were divided into three groups: a normal group (36 mice), an ethionine group (46 mice), and an ethionine+SAM group (44 mice). Phosphate-buffered saline (PBS), ethionine, and ethionine+SAM were respectively injected intraperitoneally on embryonic day 7.5 (E7.5), and the mice were sacrificed on E10.5. Embryonic tissues were collected, and the morphology of embryos in each group was observed under a stereomicroscope. The interaction between ethionine and MAT2A was analyzed using Autodock software. The expression levels of MAT2A, β-catenin, ZEB1, and EMT-related proteins in the brain tissues of embryos from the three groups were measured using immunofluorescence, immunohistochemistry, Western blotting, enzyme-linked immunosorbent assay (ELISA), and real-time quantitative polymerase chain reaction (RT-qPCR). Variance analysis was used for intergroup comparisons.Results:(1) Autodock analysis results showed that MAT2A binds to ethionine through covalent bonds, exhibiting a complementary effect, thereby accelerating the expression of MAT2A. (2) After successful construction of the NTD model, normal embryos were plump with well-developed brains. NTD embryos showed delayed development, obvious anencephaly, unclosed neural tubes, and asymmetry. (3) The levels of SAM and SAH in the embryonic tissues of the ethionine group were significantly lower than those in the normal group (1 737.56±95.64 vs. 872.33±205.11, and 89.17±9.50 vs. 51.25±9.48, respectively). The SAM and SAH levels in the ethionine+SAM group was 1 197.00±222.27 and 66.61±12.25, significantly higher than those in the ethionine group ( P<0.017). Compared with the normal group and the ethionine+SAM group, the expression of MAT2A mRNA in the embryonic brain tissue of the ethionine group was significantly upregulated (1.00±0.00, 1.59±0.52, and 2.42±0.53, respectively, F=49.64, P<0.001; pairwise comparisons between groups P<0.017). (4) Compared with the normal group, the expression of Ctnnb1 in the ethionine group was reduced, and the expression of Ctnnb1 in the ethionine+SAM group was higher than that in the ethionine group (1.00±0.00, 0.38±0.16, and 0.76±0.10, respectively, F=149.03, P<0.001; pairwise comparisons between groups P<0.017). (5) The expression of ZEB1 in the ethionine group was higher than that in the normal group and the ethionine+SAM group (2.91±0.55, 1.00±0.00, and 1.61±0.20, respectively, F=150.01, P<0.001; pairwise comparisons between groups P<0.017). (6) The expression levels of E-cadherin and Vimentin in the ethionine group were lower than those in the normal group. In contrast, the expression of N-cadherin was higher than that in the normal group. After SAM supplementation, the expression levels of E-cadherin and Vimentin were upregulated, and the expression level of N-cadherin was downregulated (0.54±0.12, 1.00±0.00, and 0.72±0.14, respectively, F=87.44; 0.53±0.17, 1.00±0.00, and 0.76±0.09, F=87.44; 3.11±0.53, 1.00±0.00, and 2.13±0.56, F=95.54; all P<0.001; pairwise comparisons within the same index group P<0.017]). Conclusions:Ethionine promotes the expression of MAT2A, leading to reduced SAM production. Ethionine regulates the level of ZEB1 by increasing MAT2A and inhibits the EMT process to interfere with methionine cycle metabolism, ultimately resulting in NTD.

Objective:To investigate the risk factors for patients with hypertriglyceridemia-related acute pancreatitis (HTG-AP) developing into severe acute pancreatitis or experiencing organ failure.Methods:This retrospective cohort study collected clinical data from 2 429 patients diagnosed with acute pancreatitis from five hospitals in China between January 2019 and December 2023 using a pre-designed data collection form. The cohort included 1 516 males and 913 females,with an age of (50.2±16.5)years(range: 11 to 99 years). Among them,353 patients (16.1%) had HTG-AP,while 1 846 (83.9%) had non-HTG-AP. HTG-AP was defined as serum triglyceride levels>500 mg/dl with other etiologies excluded. Intergroup comparisons were performed using t-tests,Mann-Whitney U test or χ2 tests,respectively. Univariate and multivariate logistic regression analyses were conducted to assess risk factors for severe acute pancreatitis after adjusting for potential confounders,and a predictive model was developed and validated. Results:Compared with other etiologies,HTG-AP patients had a higher risk of progressing to SAP ( OR=1.415,95% CI: 0.866 to 2.312, P=0.017) and organ failure ( OR=1.256,95% CI: 1.015 to 1.554, P=0.036). Among HTG-AP patients,risk factors for SAP included body mass index ( OR=1.856,95% CI: 1.742 to 1.987, P=0.033),fasting blood glucose ( OR=1.128,95% CI: 1.036 to 1.229, P=0.006),white blood cell count( OR=1.162,95% CI: 1.055 to 1.281, P=0.002),and the presence of pleural effusion ( OR=13.151,95% CI: 4.330 to 19.946, P<0.01). A nomogram prediction model for SAP in HTG-AP was constructed based on these risk factors,demonstrating good discriminative ability with area under the curve values of 0.877 in the training set and 0.894 in the validation set,along with satisfactory calibration. Conclusions:HTG-AP patients are at higher risk of developing SAP and organ failure. The risk prediction model incorporating body mass index,fasting blood glucose,white blood cell count,and pleural effusion shows good predictive value for SAP.

Objective:To evaluate the efficacy and safety of the pegaspargase, etoposide, methotrexate, and dexamethasone (PEMD) regimen in patients with early-stage nonupper respiratory digestive tract or advanced extranodal natural killer/T-cell lymphoma (ENKTL) .Methods:This retrospective analysis included 38 patients with newly diagnosed early-stage non-upper respiratory digestive tract or advanced ENKTL who received PEMD regimen for induction chemotherapy at the First Affiliated Hospital of Nanjing Medical University from January 2016 to December 2022. Survival outcomes and prognostic factors were examined by Kaplan-Meier, and the Log-rank test was used to compare survival.Results:The study population had a median age of 48 years (range, 26-72 years) and included 30 males (78.9%) and 8 females (21.1%). 7 patients’ age >60 years (18.4%). The Eastern Cooperative Oncology Group (ECOG) performance score was >1 in 7 patients (18.4%) ; 20 patients (52.6%) had elevated lactate dehydrogenase levels; and 37 patients (97.4%) exhibited extranodal involvement. Using the Ann Arbor staging system, 37 patients (97.4%) were classified as stage Ⅲ-Ⅳ. The median number of treatment cycles was 5 (1-6), and the median follow-up duration was 60 months (24 - 101 months). Interim efficacy assessment revealed an overall response rate of 52.7%. At 2 and 4 years, the progression-free survival (PFS) rates were 34.2% (95% CI 22.0%-53.2%) and 25.5% (95% CI 14.7%-44.4%), respectively, and the overall survival rates were 50.0% (95% CI 36.4%-68.7%) and 45.5% (95% CI 31.4%-65.7%), respectively. The risk factors for worse PFS were ECOG performance score >1 [ HR=3.711 (95% CI 1.494-9.218), P=0.005]; bone marrow infiltration [ HR=2.251 (95% CI 1.026 - 4.938), P=0.043]; and Prognostic Index for Natural Killer/T-Cell Lymphoma score of 3 - 5 [ HR=2.350 (95% CI 1.009 - 5.476), P=0.048]. Multivariate analysis identified ECOG performance score >1 as an independent risk factor for PFS [ HR=7.971 (95% CI 2.222 - 28.591), P=0.001]. The main adverse effect was anemia in 31 patients (81.6%) . Conclusion:The PEMD regimen was safe and effective for patients with newly diagnosed early-stage non-upper respiratory digestive tract or advanced ENKTL.

Objective To explore the effects and mechanism of calycosin-7-glucoside(CG)on lipopolysaccharide(LPS)-induced inflammatory injury in BV-2 cells and in a mouse model of neuroinflammation.Methods An in vitro neuroinflammation model was induced by LPS stimulation of BV-2 cells.BV-2 cells were divided into blank(CON),model(LPS),dexamethasone(DEX),and low-and high-dose CG(CG 10 μmol/L,CG 20 μmol/L,respectively)groups.The cell viability in each group was detected by Cell Counting Kit-8 assay,interleukin(IL)-6 and tumor necrosis factor(TNF)-α levels in the supernatant were detected by enzyme-linked immunosorbent assay(ELISA),and nitric oxide levels were detected using the Griess method.LPS was also used to induce neuroinflammation in mice in vivo.The mice were then divided randomly into blank(CON),model(LPS),aspirin,and low-and high-dose CG(CG 5 mg/kg,CG 10 mg/kg,respectively)groups.Pathological changes in the hippocampus were detected by hematoxylin/eosin staining.Serum levels of IL-6 and TNF-α were detected by ELISA,polarization of microglia was detected by immunofluorescence staining,and protein expression levels of Toll-like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),nuclear factor κB(NF-κB,P65)and phosphorylated-NF-κB(p-P65)in the cortex were detected by Western blot.Results CG alone or in combination with LPS in the concentration range of 2.5～160 μmol/L had no significant toxicity in BV-2 cells in vitro,compared with the CON group(P＞0.05).IL-6,TNF-α,and NO levels in the cell supernatant were increased in the LPS group compared with the CON group(P＜0.01),but were significantly reduced by CG(P＜0.05,P＜0.01).Hippocampal neurons were arranged loosely and disordered in the LPS group in vivo,compared with the CON group,and nuclear pyknosis was observed.Serum levels of IL-6 and TNF-α were increased(P＜0.05,P＜0.01).The number of ionized calcium binding adaptor molecule 1(Iba1)/inducible nitric oxide synthase(iNOS)cells was increased(P＜0.01),the number of CD206/Iba1 cells was decreased(P＜0.01),and expression levels of TLR4,MyD88,and p-P65 protein in the cortex were increased(P＜0.05).Compared with the LPS group,CG improved the pathological damage to the hippocampus and inhibited serum levels of IL-6 and TNF-α(P＜0.01).CG also decreased the number of iNOS/Iba1 cells,increased the number of CD206/Iba1 cells(P＜0.05,P＜0.01),and significantly down-regulated TLR4,MyD88,and p-P65 protein levels in the cortex(P＜0.05).Conclusions CG can ameliorate neuroinflammation in mice by suppressing the TLR4/MyD88/NF-κB pathway.