ObjectiveTo investigate the effects and underlying mechanisms of Shenqi Wenfei prescription （SQWF） on chronic obstructive pulmonary disease （COPD）. MethodsA rat model of COPD with lung Qi deficiency was established using lipopolysaccharide （LPS） combined with cigarette smoke. Forty-eight SD rats were randomly divided into a blank group， a model group， low-， medium-， and high-dose SQWF groups （2.835， 5.67， 11.34 g·kg^-1）， and a Yupingfeng group （1.35 g·kg^-1）. Drug administration began on day 29 after modeling and continued for 2 weeks. The general condition of the rats was observed， and the lung function in each group was assessed. Hematoxylin-eosin （HE） staining was used to observe pathological changes in lung tissue. The proportion of inflammatory cells in bronchoalveolar lavage fluid （BALF） was measured. Apoptosis in lung tissue was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （TUNEL） staining. The release level of lactate dehydrogenase （LDH） in BALF was detected by a microplate assay. Reactive oxygen species （ROS） levels in lung tissue were detected using fluorescent probes. The levels of malondialdehyde （MDA）， total superoxide dismutase （SOD）， and reduced glutathione （GSH） in BALF were measured by biochemical methods. Ultrastructural changes in lung cells were observed via transmission electron microscopy. Double immunofluorescence staining was performed to detect the expression of thioredoxin-interacting protein （TXNIP） and nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3） in lung tissue. Western blot analysis was used to detect the protein expression of TXNIP， NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， cysteinyl aspartate-specific protease-1 （Caspase-1）， Caspase-1 p20， gasdermin D （GSDMD）， GSDMD N-terminal active fragment （GSDMD-N）， interleukin-1β （IL-1β）， and IL-18 in lung tissue. Serum IL-1β and IL-18 levels were measured by ELISA. ResultsCompared with the blank group， the model group showed lassitude， fatigue， tachypnea， and audible phlegm sounds， and lung function significantly declined （P0.01）. Pulmonary emphysema and inflammatory cell infiltration were obvious. The level of inflammatory cells in BALF increased significantly （P0.05）. The number of TUNEL-positive cells increased （P0.01）. Levels of LDH， ROS， and MDA in BALF increased significantly （P0.01）， while GSH and SOD activities decreased significantly （P0.01）. Lung tissue cells showed irregular morphology， swollen mitochondria， disrupted cell membranes， and abundant vesicles， i.e.， pyroptotic bodies. Protein levels of TXNIP， NLRP3， ASC， Caspase-1， Caspase-1 p20， GSDMD， GSDMD-N， IL-1β， and IL-18 in lung tissue were significantly elevated （P0.01）， and serum IL-1β and IL-18 levels also increased significantly （P0.01）. Compared with the model group， each medication group showed alleviation of qi deficiency symptoms and improved lung function （P0.01）. Pulmonary emphysema and inflammatory cell infiltration were reduced. Inflammatory cell levels decreased （P0.05）. The number of TUNEL-positive cells decreased significantly （P0.01）. Levels of LDH， ROS， and MDA decreased significantly （P0.05）， while GSH and SOD activities significantly increased （P0.01）. Morphological and structural damage in lung tissue was improved to varying degrees. Protein levels of TXNIP， NLRP3， ASC， Caspase-1， Caspase-1 p20， GSDMD， GSDMD-N， IL-1β， and IL-18 in lung tissue significantly decreased （P0.01）， and serum IL-1β and IL-18 levels also decreased significantly （P0.05）. ConclusionSQWF can improve lung function and alleviate inflammatory responses in COPD rats. Its mechanism may be related to regulating the ROS/TXNIP/NLRP3 pathway and inhibiting pyroptosis.

ObjectiveTo investigate the effects and underlying mechanisms of Shenqi Wenfei prescription （SQWF） on chronic obstructive pulmonary disease （COPD）. MethodsA rat model of COPD with lung Qi deficiency was established using lipopolysaccharide （LPS） combined with cigarette smoke. Forty-eight SD rats were randomly divided into a blank group， a model group， low-， medium-， and high-dose SQWF groups （2.835， 5.67， 11.34 g·kg^-1）， and a Yupingfeng group （1.35 g·kg^-1）. Drug administration began on day 29 after modeling and continued for 2 weeks. The general condition of the rats was observed， and the lung function in each group was assessed. Hematoxylin-eosin （HE） staining was used to observe pathological changes in lung tissue. The proportion of inflammatory cells in bronchoalveolar lavage fluid （BALF） was measured. Apoptosis in lung tissue was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （TUNEL） staining. The release level of lactate dehydrogenase （LDH） in BALF was detected by a microplate assay. Reactive oxygen species （ROS） levels in lung tissue were detected using fluorescent probes. The levels of malondialdehyde （MDA）， total superoxide dismutase （SOD）， and reduced glutathione （GSH） in BALF were measured by biochemical methods. Ultrastructural changes in lung cells were observed via transmission electron microscopy. Double immunofluorescence staining was performed to detect the expression of thioredoxin-interacting protein （TXNIP） and nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3） in lung tissue. Western blot analysis was used to detect the protein expression of TXNIP， NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， cysteinyl aspartate-specific protease-1 （Caspase-1）， Caspase-1 p20， gasdermin D （GSDMD）， GSDMD N-terminal active fragment （GSDMD-N）， interleukin-1β （IL-1β）， and IL-18 in lung tissue. Serum IL-1β and IL-18 levels were measured by ELISA. ResultsCompared with the blank group， the model group showed lassitude， fatigue， tachypnea， and audible phlegm sounds， and lung function significantly declined （P0.01）. Pulmonary emphysema and inflammatory cell infiltration were obvious. The level of inflammatory cells in BALF increased significantly （P0.05）. The number of TUNEL-positive cells increased （P0.01）. Levels of LDH， ROS， and MDA in BALF increased significantly （P0.01）， while GSH and SOD activities decreased significantly （P0.01）. Lung tissue cells showed irregular morphology， swollen mitochondria， disrupted cell membranes， and abundant vesicles， i.e.， pyroptotic bodies. Protein levels of TXNIP， NLRP3， ASC， Caspase-1， Caspase-1 p20， GSDMD， GSDMD-N， IL-1β， and IL-18 in lung tissue were significantly elevated （P0.01）， and serum IL-1β and IL-18 levels also increased significantly （P0.01）. Compared with the model group， each medication group showed alleviation of qi deficiency symptoms and improved lung function （P0.01）. Pulmonary emphysema and inflammatory cell infiltration were reduced. Inflammatory cell levels decreased （P0.05）. The number of TUNEL-positive cells decreased significantly （P0.01）. Levels of LDH， ROS， and MDA decreased significantly （P0.05）， while GSH and SOD activities significantly increased （P0.01）. Morphological and structural damage in lung tissue was improved to varying degrees. Protein levels of TXNIP， NLRP3， ASC， Caspase-1， Caspase-1 p20， GSDMD， GSDMD-N， IL-1β， and IL-18 in lung tissue significantly decreased （P0.01）， and serum IL-1β and IL-18 levels also decreased significantly （P0.05）. ConclusionSQWF can improve lung function and alleviate inflammatory responses in COPD rats. Its mechanism may be related to regulating the ROS/TXNIP/NLRP3 pathway and inhibiting pyroptosis.

ObjectiveTo evaluate the clinical efficacy and safety of Jingmaiyan granules （composed of Lonicerae Japonicae Flos， Sedi Herba， Paeoniae Radix Rubra， Moutan Cortex， Rhei Radix et Rhizoma Praeparata， and Glycyrrhizae Radix et Rhizoma） combined with external application of Jinhuang Ointment in treating acute-stage blood heat stasis type superficial thrombophlebitis （ST） of lower extremities， and to explore their effects on hemorheology and serum inflammatory factors. MethodsA randomized， double-blind， placebo-controlled clinical trial was conducted. A total of 124 patients with lower extremity ST were randomized into two groups（62 cases in each group）. The control group received external application of Jinhuang ointment and oral placebo treatment， while the observation group received external application of Jinhuang ointment and oral Jingmaiyan granules. Both groups were treated for 2 weeks. The clinical symptom scores， therapeutic efficacy of traditional Chinese medicine （TCM） syndrome， pain visual analog scale （VAS） scores， hemorheological indices ［including whole blood high-shear， medium-shear， and low-shear viscosity， as well as plasma viscosity （PV）］， and inflammatory factors ［C-reactive protein （CRP）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）］ were compared before and after treatment. ResultsAfter 2 weeks of treatment， the total effective rate in the observation group （98.3%， 60/62） was significantly higher than that in the control group （83.8%， 52/62）， with a statistically significant difference （Z=3.512 1， P<0.05）. Compared with pre-treatment scores， the scores for skin color， skin temperature， swelling， pain， and cord or nodules were significantly reduced in both groups （P<0.05）， with more pronounced improvement in the observation group （P<0.05）. Additionally， compared with pre-treatment levels， the whole blood viscosity （low-， medium-， and high-shear） significantly improved in both groups after treatment （P<0.05）， with more marked improvement in the observation group （P<0.05）. Furthermore， the plasma viscosity， CRP， IL-6， and TNF-α levels were significantly reduced in both groups after treatment （P<0.05）， with more pronounced improvement observed in the observation group （P<0.05）. ConclusionThe combination of external application of Jinhuang ointment and oral Jingmaiyan granules effectively improves clinical symptoms， hemorheological abnormalities， and inflammatory responses in patients with acute stage blood heat stasis type ST of lower extremities. The treatment is safe and holds clinical promotion value.

ObjectiveTo establish an animal model of atrial fibrillation（AF） that accurately reflects the phlegm-heat and blood stasis（TRYZ） pathogenesis in traditional Chinese medicine. MethodsForty SPF-grade SD rats were randomly assigned using a random number table to the following groups：the control group， the TRYZ+AF group，the AF group and the TRYZ group， with ten rats in each group. The TRYZ+AF and TRYZ groups underwent a high-fat diet combined with intraperitoneal lipopolysaccharide（LPS） injection to simulate the pathological alterations of TRYZ syndrome. Groups TRYZ+AF and AF were induced with acetylcholine-calcium chloride（Ach-CaCl₂） via caudal vein injection to induce AF. The control group received no intervention and was maintained under normal conditions. The modeling period lasted 3 weeks. Electrocardiography was used to assess AF episodes and duration， echocardiography evaluated left atrial dimensions and cardiac function， fully automated biochemical analyzer measured the levels of total cholesterol（TC）， triglycerides（TG）， high-density lipoprotein cholesterol（HDL-C） and low-density lipoprotein cholesterol（LDL-C）， hemoreometer analyzed the whole blood viscosity， plasma viscosity， and whole blood reduced viscosity， a coagulation analyzer assessed prothrombin time（PT）， activated partial thromboplastin time（APTT）， thrombin time（TT）， and fibrinogen（FIB）， enzyme-linked immunosorbent assay（ELISA） was used to determine the levels of C-reactive protein（CRP）， interleukin（IL）-1β， IL-6， IL-17， tumour necrosis factor（TNF）-α， matrix metalloproteinase-9（MMP-9）， galectin-3（Gal-3）， Collagen Ⅰ， and α-smooth muscle actin（α-SMA）. Hematoxylin-eosin（HE） staining and Masson's trichrome staining were used to analyze pathological changes in atrial myocardium， Western blot was employed to detect MMP-9， Collagen Ⅰ and α-SMA protein expression in myocardial tissue， real-time quantitative polymerase chain reaction（Real-time PCR） evaluated fibrous factor gene expression levels. Changes in the TRYZ syndrome were assessed via body weight， tongue color［red（R）， green（G）， and blue（B）］， and rectal temperature. Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was employed to detect differential metabolites between the control group and the TRYZ+AF group. ResultsFollowing three weeks of sustained modeling， compared with the control group， rats in the TRYZ+AF and the TRYZ groups exhibited reduced body weight， dry faeces， elevated rectal temperature， dark red tongue， decreased RGB values on the tongue surface， and markedly elevated TC and LDL-C levels（P<0.05， P<0.01）. The TRYZ+AF， TRYZ， and AF groups exhibited significantly decreased TT， APTT and PT， along with markedly elevated whole blood viscosity and FIB（P<0.05， P<0.01）. Rats in the TRYZ+AF and AF groups exhibited AF rhythm， markedly decreased heart rate， prolonged RR intervals， enlarged left atrium， and significantly reduced ejection fraction and shortening fraction（P<0.05， P<0.01）. Serum levels of CRP， IL-1β， IL-6， IL-17， TNF-α， MMP-9， Gal-3， Collagen Ⅰ， and α-SMA were elevated in rats from the TRYZ+AF， TRYZ， and AF groups compared to the control group， with the most pronounced increase observed in the TRYZ+AF group（P<0.05， P<0.01）. Histopathology revealed that the collagen fiber deposition in the atrial of rats in the TRYZ+AF， TRYZ and AF groups was higher than that in the control group（P<0.05， P<0.01）. Western blot and Real-time PCR results further demonstrated that the protein and mRNA expression levels of MMP-9， Collagen Ⅰ and α-SMA in the myocardial tissue of the TRYZ+AF group were higher than those in the other three groups（P<0.05， P<0.01）. Metabolomic analysis revealed 173 differentially expressed metabolites in the TRYZ+AF group and the control group， primarily enriched in pathways such as glycerophospholipid metabolism and glycolysis/gluconeogenesis. ConclusionThis study successfully establishes a rat model of AF integrated with the TRYZ syndrome， demonstrating the pathological process where the interactions of phlegm， heat and stasis jointly trigger tremor， this provides a reliable experimental tool for in-depth research into the biological basis of this disease syndrome.

ObjectiveTo establish fingerprint profiles and a quantitative determination method for Lycii Cortex， providing a scientific basis for the formulation of quality standards for Lycii Cortex and its roasted products. MethodsHigh performance liquid chromatography（HPLC） was developed for the quantitative method for determining kukoamine B in Lycii Cortex and its roasted products on an Alphasil XD-C₁₈ CH column（4.6 mm×250 mm， 5 μm）. HPLC fingerprint profiles were established for 10 batches of Lycii Cortex and its roasted products， and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used to identify the common peaks based on reference standards， literature and MS information. Quality evaluation indicators included yield of decoction pieces， appearance properties， content of kukoamine B， and fingerprint profiles. The temperature and time of the roasting process were investigated to select the optimal preparation process， which was then verified. Additionally， chemical pattern recognition was combined to assess the differences in the chemical composition of Lycii Cortex before and after roasting， as well as among samples from different origins. ResultsQuantitative analysis indicated that the contents of kukoamine B in Lycii Cortex and its roasted products were 0.35%-5.51% and 0.24%-4.15%， respectively. The transfer rate of kukoamine B was 58.6%-78.9% after roasting. The fingerprint profile analysis demonstrated that the method established in this study effectively separated kukoamine B from other components in the samples and distinctly differentiated it from its impurity peak， cis-N-caffeoylputrescine. The HPLC fingerprint profiles of Lycii Cortex and its roasted products showed high similarity（all above 0.95）， with 7 common peaks identified and five common components， including kukoamine B， cis-N-caffeoylputrescine， N-coumaroyl tyramine， feruloyltyramine， and glucosyringic acid， confirmed. Process optimization confirmed that baking at 110 ℃ for 20 min was a stable and feasible method for roasting Lycii Cortex. Principal component analysis and cluster analysis showed that there was little difference in the chemical composition between raw and roasted Lycii Cortex， but the quality of Lycii Cortex from different origins differed greatly. ConclusionThis study successfully established the fingerprint profiles and a quantitative method for the effective component kukoamine B in Lycii Cortex and roasted Lycii Cortex. The qualitative and quantitative analyses clarified that the impact of the roasting process on the chemical composition of Lycii Cortex was less significant than the variations due to its geographical origin. The findings of this study offer a reference for the development of quality evaluation methods and the establishment of quality standards for Lycii Cortex and its processed products.

ObjectiveTo investigate the influencing factors for traditional Chinese medicine （TCM） syndromes of primary liver cancer， and to provide a theoretical basis for the TCM syndrome differentiation and standardized treatment of liver cancer. MethodsTCM syndrome differentiation was performed for 415 patients who were admitted to Shanxi Institute of Traditional Chinese Medicine and were diagnosed with primary liver cancer based on pathological or clinical examinations from January 2019 to December 2023. The chi-square test was used for comparison of categorical data between groups， and the unordered polytomous logistic regression model was used to investigate the influencing factors for TCM syndromes of liver cancer. ResultsThe common initial symptoms of the 415 patients with primary liver cancer included pain in the liver area （31.81%）， abdominal distension （25.30%）， abdominal pain （15.18%）， and weakness （13.98%）， and the main clinical symptoms included poor appetite （70.84%）， fatigue （69.16%）， pain in the liver area （67.47%）， poor sleep （59.04%）， abdominal distension （53.01%）， and constipation （52.53%）. There were significant differences in TCM syndromes between patients with different sexes， courses of the disease， clinical stages， Child-Pugh classes， presence or absence of intrahepatic and extrahepatic metastasis， and presence or absence of transcatheter arterial chemoembolization （TACE） and radiofrequency ablation （all P<0.05）. The logistic regression analysis showed that male sex was a risk factor for damp-heat accumulation （odds ratio ［OR］=2.036， P=0.048） and the syndrome of spleen-kidney Yang deficiency （OR=5.240， P<0.001）； a course of disease of<1 year was a risk factor for damp-heat accumulation （OR=2.837， P=0.004） and syndrome of Qi stagnation and blood stasis （OR=2.317， P=0.021）， but it was a protective factor against syndrome of spleen-kidney Yang deficiency （OR=0.385， P=0.005）； Child-Pugh class A/B was a protective factor against liver-kidney Yin deficiency （OR=0.079， P<0.001）； intrahepatic metastasis was a risk factor for liver-kidney Yin deficiency （OR=5.117， P=0.003） and syndrome of spleen-kidney Yang deficiency （OR=3.303， P=0.010）； TACE was a protective factor against liver-kidney Yin deficiency （OR=0.171， P<0.001） and syndrome of spleen-kidney Yang deficiency （OR=0.138， P<0.001）； radiofrequency ablation was a risk factor for damp-heat accumulation （OR=4.408， P<0.001） and liver-kidney Yin deficiency （OR=32.036， P<0.001）. ConclusionSex， course of disease， Child-Pugh class， intrahepatic metastasis， TACE， and radiofrequency ablation are the main influencing factors for TCM syndromes of liver cancer.

ObjectiveTo explore the mechanism by which modified Shaofu Zhuyutang inhibits angiogenesis in endometriosis via the vascular endothelial growth factor （VEGF）/phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/endothelial nitric oxide synthase （eNOS）-nitric oxide （NO） signaling pathway. MethodsEighty-four female SD rats were randomly assigned into blank， sham operation， model， positive control （gestrinone， 0.25 mg·kg^-1）， high-， medium-， and low-dose （30， 15， 7.5 g·kg^-1， respectively） traditional Chinese medicine （TCM， modified Shaofu Zhuyutang） groups. A rat model of endometriosis was established by the autotransplantation method. After successful modeling， rats in the drug intervention groups were administrated with corresponding agents by gavage， and those in the blank， sham operation， and model groups received an equal volume of distilled water. After 28 days of gavage， rats were administrated with oxytocin， and the number and latency period of writhing responses were observed. Serum samples from each group， ectopic lesions from modeling groups， and uteri from blank and sham operation groups were collected. Hematoxylin-eosin staining was used to observe the pathological morphology of endometriotic lesions. Immunohistochemistry was employed to observe the expression of angiogenesis-specific markers cluster of differentiation 34 antigen （CD34） and friend leukemia virus integration-1 （FLI-1）. Enzyme-linked immunosorbent assay and the nitrate reductase method were employed to determine the serum levels of VEGF and NO， respectively. Western blot was employed to measure the protein levels of VEGF， PI3K， phosphorylated PI3K （p-PI3K）， Akt， phosphorylated Akt （p-Akt）， eNOS， and phosphorylated eNOS （p-eNOS）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was conducted to determine the mRNA levels of VEGF， PI3K， Akt， and eNOS. ResultsThe blank group and the sham operation group had no significant changes in the number and latency period of writhing responses， serum VEGF and NO levels， protein levels of VEGF， p-PI3K/PI3K， p-Akt/Akt， and p-eNOS/eNOS， and mRNA levels of VEGF， PI3K， Akt， and eNOS. The model group showed an increase in the number and a reduction in the latency period of writhing responses， enlargement of ectopic endometrial tissue in the abdominal wall， with stromal hyperplasia， glandular dilation， and increased vasculature. In addition， the modeling led to increased positive expression of CD34 and FLI-1， elevated serum VEGF and NO levels， and up-regulated protein levels of VEGF， p-PI3K/PI3K， p-Akt/Akt， and p-eNOS/eNOS and mRNA levels of VEGF， PI3K， Akt， and eNOS （P<0.01）. Compared with the model group， the gestrinone and high-， medium-， and low-dose TCM groups showed a significant reduction in the number of writhing responses， a significant prolongation of the latency period， reduced ectopic endometrial tissue in the abdominal wall， alleviated pathological damage， and reduced positive expression of CD34 and FLI-1. The gestrinone group and the high- and medium-dose TCM groups showed lowered serum VEGF and NO levels as well as down-regulated protein levels of VEGF， p-PI3K/PI3K， p-Akt/Akt， and p-eNOS/eNOS and mRNA levels of VEGF， PI3K， Akt， and eNOS. Moreover， the low-dose TCM group showed reductions in the serum VEGF level， the protein levels of VEGF， p-PI3K/PI3K， p-Akt/Akt， and p-eNOS/eNOS， and the mRNA levels of VEGF and eNOS （P<0.05， P<0.01）. ConclusionModified SShaofu Zhuyutang can inhibit angiogenesis in endometriosis by antagonizing the abnormal activation of the VEGF/PI3K/Akt/eNOS-NO signaling pathway， thereby preventing the occurrence， development， and deterioration of endometriosis.

BackgroundPost-traumatic stress symptoms （PTSS） is highly prevalent in adolescents who have experienced earthquake， which seriously threatens their physical and mental health， yet there is currently a lack of research on the effects of loneliness， social support and social media use on PTSS among post-earthquake adolescents. ObjectiveTo assess the PTSS among adolescents experiencing MS6.0 Luxian， Sichuan， earthquake on 16 September 2021， and to investigate the effects of loneliness， social support and social media use on PTSS， so as to provide references for the intervention of PTSS among post-earthquake adolescents. MethodsOn November 12， 2021， simple random sampling technique was used to select 2 522 post-earthquake adolescents in Luxian county of Luzhou city in Sichuan province. All subjects were assessed using Post-Traumatic Stress Disorder Checklist for DSM-5 （PCL-5）， Multidimensional Scale of Perceived Social Support （MSPSS）， Short-form UCLA Loneliness Scale （ULS-3） and Social Media Use Scale （SM-10）. Binary Logistic regression was used to determine the factors influencing PTSS among post-earthquake adolescents. ResultsPTSS was detected in 91 （3.61%） adolescents. Binary Logistic regression revealed that perceived social support from family members （OR=0.926， 95% CI： 0.879~0.976） was a protective factor for PTSS among post-earthquake adolescents. Lack of companionship （OR=1.764， 95% CI： 1.141~2.727）， feeling isolated （OR=2.037， 95% CI： 1.282~3.236）， and viewing negative emotional response of disaster victims through social media （OR=1.615， 95% CI： 1.291~2.020） were risk factors for PTSS among post-earthquake adolescents. ConclusionLack of companionship， feeling isolated， and viewing negative emotional response of disaster victims through social media pose a negative impact on PTSS among post-earthquake adolescents， while perceived social support from family members exert a positive impact on PTSS among post-earthquake adolescents. ［Funded by Humanity and Social Science Youth foundation of Ministry of Education of China （number， 22YJC190019）； Natural Science Foundation of Sichuan Province （number， 2023NSFSC1486）］

ObjectiveTo explore the mechanism by which modified Shaofu Zhuyutang inhibits angiogenesis in endometriosis via the vascular endothelial growth factor （VEGF）/phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/endothelial nitric oxide synthase （eNOS）-nitric oxide （NO） signaling pathway. MethodsEighty-four female SD rats were randomly assigned into blank， sham operation， model， positive control （gestrinone， 0.25 mg·kg^-1）， high-， medium-， and low-dose （30， 15， 7.5 g·kg^-1， respectively） traditional Chinese medicine （TCM， modified Shaofu Zhuyutang） groups. A rat model of endometriosis was established by the autotransplantation method. After successful modeling， rats in the drug intervention groups were administrated with corresponding agents by gavage， and those in the blank， sham operation， and model groups received an equal volume of distilled water. After 28 days of gavage， rats were administrated with oxytocin， and the number and latency period of writhing responses were observed. Serum samples from each group， ectopic lesions from modeling groups， and uteri from blank and sham operation groups were collected. Hematoxylin-eosin staining was used to observe the pathological morphology of endometriotic lesions. Immunohistochemistry was employed to observe the expression of angiogenesis-specific markers cluster of differentiation 34 antigen （CD34） and friend leukemia virus integration-1 （FLI-1）. Enzyme-linked immunosorbent assay and the nitrate reductase method were employed to determine the serum levels of VEGF and NO， respectively. Western blot was employed to measure the protein levels of VEGF， PI3K， phosphorylated PI3K （p-PI3K）， Akt， phosphorylated Akt （p-Akt）， eNOS， and phosphorylated eNOS （p-eNOS）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was conducted to determine the mRNA levels of VEGF， PI3K， Akt， and eNOS. ResultsThe blank group and the sham operation group had no significant changes in the number and latency period of writhing responses， serum VEGF and NO levels， protein levels of VEGF， p-PI3K/PI3K， p-Akt/Akt， and p-eNOS/eNOS， and mRNA levels of VEGF， PI3K， Akt， and eNOS. The model group showed an increase in the number and a reduction in the latency period of writhing responses， enlargement of ectopic endometrial tissue in the abdominal wall， with stromal hyperplasia， glandular dilation， and increased vasculature. In addition， the modeling led to increased positive expression of CD34 and FLI-1， elevated serum VEGF and NO levels， and up-regulated protein levels of VEGF， p-PI3K/PI3K， p-Akt/Akt， and p-eNOS/eNOS and mRNA levels of VEGF， PI3K， Akt， and eNOS （P<0.01）. Compared with the model group， the gestrinone and high-， medium-， and low-dose TCM groups showed a significant reduction in the number of writhing responses， a significant prolongation of the latency period， reduced ectopic endometrial tissue in the abdominal wall， alleviated pathological damage， and reduced positive expression of CD34 and FLI-1. The gestrinone group and the high- and medium-dose TCM groups showed lowered serum VEGF and NO levels as well as down-regulated protein levels of VEGF， p-PI3K/PI3K， p-Akt/Akt， and p-eNOS/eNOS and mRNA levels of VEGF， PI3K， Akt， and eNOS. Moreover， the low-dose TCM group showed reductions in the serum VEGF level， the protein levels of VEGF， p-PI3K/PI3K， p-Akt/Akt， and p-eNOS/eNOS， and the mRNA levels of VEGF and eNOS （P<0.05， P<0.01）. ConclusionModified SShaofu Zhuyutang can inhibit angiogenesis in endometriosis by antagonizing the abnormal activation of the VEGF/PI3K/Akt/eNOS-NO signaling pathway， thereby preventing the occurrence， development， and deterioration of endometriosis.

BACKGROUND:Rheumatoid arthritis is a condition that affects the entire immune system in the body and is known for causing inflammatory hyperplasia in the joints and destruction of articular cartilage.The pathogenesis of rheumatoid arthritis is still unclear;therefore,there is an urgent need to discover new highly sensitive and specific diagnostic biomarkers. OBJECTIVE:To identify and screen key genes in the synovium of rheumatoid arthritis patients using bioinformatics techniques and machine learning algorithms and to construct and validate a rheumatoid arthritis prediction model. METHODS:Three datasets containing synovial tissue samples from rheumatoid arthritis patients(GSE77298,GSE55235,GSE55457)were downloaded from the Gene Expression Omnibus(GEO)database.GSE77298 and GSE55235 were used as the training set,while GSE55457 served as the test set,with a total of 66 samples,including 39 samples from rheumatoid arthritis patients and 27 normal synovial samples.Differentially expressed genes in the training set were selected using R language,and then the weighted gene co-expression network analysis was used to modularize the genes in the training set.The most relevant module was selected,and feature genes within this module were identified.Differentially expressed genes and the feature genes from the module were intersected for the subsequent machine learning analysis.Three machine learning methods,namely the least absolute shrinkage and selection operator algorithm,support vector machine with recursive feature elimination,and random forest algorithm,were employed to further analyze the intersected genes and identify the hub genes.The hub genes obtained from these three machine learning algorithms were intersected again to obtain the key genes in the synovium of rheumatoid arthritis.A predictive rheumatoid arthritis model was constructed using these key genes as variables,and the risk of developing rheumatoid arthritis in patients was inferred based on the model.The receiver operating characteristic curve was used to determine the diagnostic value of the rheumatoid arthritis prediction model and its key genes. RESULTS AND CONCLUSION:Through the differential analysis,a total of 730 differentially expressed genes were identified in the training set,and 185 feature genes were identified in the weighted gene co-expression network analysis feature modules.There were 159 intersected genes obtained.There were 4 hub genes identified by the least absolute shrinkage and selection operator algorithm,11 hub genes by the support vector machine with recursive feature elimination algorithm,and 5 hub genes by the random forest algorithm.After intersection,2 key genes(TNS3 and SDC1)were obtained.Based on the two key genes,a nomogram model was constructed in the training and test sets,with good fit between the calibration prediction curve and the standard curve,and good clinical efficacy in predicting the onset of rheumatoid arthritis.These findings indicate that TNS3 and SDC1,obtained based on bioinformatics and machine learning algorithms,may become key targets for the diagnosis and treatment of rheumatoid arthritis.