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.

Objective To evaluate the clinical efficacy of a novel sleep-aid decoction in treating elderly insomnia patients with different traditional Chinese medicine (TCM) constitutional types, and its effects on neurotransmitter and inflammatory factor levels. Methods A total of 200 patients with four different TCM constitutions-peaceful, Qi-deficient, Yin-deficient, and Yang-deficient-were recruited. Peripheral blood neurotransmitter and inflammatory factor levels were measured for variations among insomnia patients across different constitutions. These patients were treated using the novel sleep-aid decoction, the effects of which were evaluated based on changes in neurotransmitters and inflammatory factors. Results Compared to the peaceful constitution group, insomnia patients with Qi-deficient, Yin-deficient, and Yang-deficient constitutions exhibited significantly elevated baseline levels of neurotransmitters (5-HT, GABA) and inflammatory factors (IL-6, TNF-α, IL-1β, CRP). Following the treatment, the Qi-deficient and Yin-deficient groups showed a marked increase in 5-HT levels, restored balance of Glu, GABA, and melatonin, and significant reductions in IL-6 and TNF-α levels. The overall effective rate was 83.5%, with optimal efficacy observed in the Qi-deficient (97.72%) and Yin-deficient (95.34%) groups. Conclusion The novel sleep-aid decoction is effective in treating insomnia in elderly patients, with the best results observed in the Qi-deficient and Yin-deficient constitution groups.
Humans ; Sleep Initiation and Maintenance Disorders/blood* ; Aged ; Male ; Female ; Drugs, Chinese Herbal/therapeutic use* ; Medicine, Chinese Traditional ; Middle Aged ; Tumor Necrosis Factor-alpha/blood* ; Sleep Aids, Pharmaceutical/therapeutic use* ; Anti-Inflammatory Agents/therapeutic use* ; Interleukin-6/blood* ; Interleukin-1beta/blood* ; Neurotransmitter Agents/blood* ; Aged, 80 and over ; C-Reactive Protein/metabolism*

OBJECTIVE: Hepatocellular carcinoma (HCC) is a leading cause of mortality worldwide. Huachansu (Cinobufacini) is active extract isolated from the dry skin of Bufo Bufo gargarizans. It has now been widely used in clinical treatment of cancer, this study is to clarify the material basis of down-regulation of thymidylate synthase (TYMS) induced by Huachansu. METHODS: Our study utilized UPLC-MS/MS to identify major bioactive components from Huachansu. Cell Counting Kit 8 (CCK-8) assay and clone formation assay were used to examine the cell viability of tumor cells. TYMS and γ-H2AX level were detected by using quantitative real-time RT-PCR and/or western blotting. Small interfering RNA (siRNA) transfection was used to explore whether inhibition of TYMS could enhance the suppressive effect of Huachansu on cell growth of HCC cells. RESULTS: In our study, firstly, we identify 21 major bioactive components from Huachansu. CCK-8 assay results showed that Huachansu and its bioactive bufadienolides (Bufalin, Bufotalin, Cinobufotalin, Desacetylcinobufagin, Arenobufagin, Telocinobufagin, and Resibufogenin) significantly inhibited the proliferation of HepG2 and SK-HEP-1 cells in a dose- and time-dependent manner. Further molecular mechanistic investigation demonstrates that Huachansu significantly suppresses thymidylate synthase (TYMS), the enzyme which provides the sole de novo source of thymidylate for DNA synthesis. The inhibition of TYMS could lead to cell-cycle block and DNA damage of HCC cells. Furthermore, we identified that Huachansu markedly increased γ-H2AX expression, which indicated the presence of DNA damage. Moreover, we confirmed that transfection of cells with small interfering RNA specific to TYMS could increase the suppressive effects of Huachansu on the HCC cells proliferation. Quantitative RT-PCR analysis showed that Huachansu treatment had no effect on the transcription level of TYMS. Furthermore, proteasomal inhibitor MG132 could block TYMS inhibition induced by Huachansu, and concomitant administration of protein synthesis inhibitor cycloheximide (CHX) with Huachansu could further suppress the protein level of TYMS, indicating that Huachansu promotes proteasome-dependent degradation of TYMS in liver cancer cells. More importantly, the bioactive bufadienolides of Huachansu such as Bufalin, Bufotalin, Cinobufotalin, Desacetylcinobufagin, Arenobufagin, Telocinobufagin, and Resibufogenin could also significantly restrain the protein level of TYMS, revealing the material basis of inhibition of TYMS exposed to Huachansu. 5-Fluorouracil (5-FU) is a TYMS inhibitor, we also evaluate the effects of the combined treatment of Huachansu with 5-FU, the results show that interactions between Huachansu and 5-FU are synergistic or antagonistic. Thus, in clinical, attention should be paid to the dosage of Huachansu in combination with 5-FU. CONCLUSION Huachansu inhibits the growth and induces DNA damage of human HCC cells through proteasome-dependent degradation of TYMS, bioactive bufadienolides are the material basis of down-regulation of TYMS induced by Huachansu.

Objective To analyze the medication law of Professor Yuan Jinsheng,a renowned TCM practitioner in China,for treating palpitations through data mining methods.Methods Clinical prescriptions for treating palpitations by Professor Yuan Jinsheng from January 2016 to May 2023 were collected.The prescriptions were screened based on inclusion and exclusion criteria,a prescription compatibility network was constructed based on R Studio 4.3.1,and the medication law of prescriptions was analyzed.Results Totally 331 prescriptions were screened,involving 180 types of Chinese materia medica,with a total frequency of 3 625.The most frequently used drugs(≥30 times)were mainly tonics.The main properties were warm and neutral,the main tastes were sweet,bitter,and pungent,which belonged to heart,spleen and lung meridians.The top 5 drugs with high correlation were tonifying,blood circulation-activating and stasis-resolving,qi-regulating,and heat-clearing.Correlation analysis reveals high-frequency drugs,which were mainly Glycyrrhizae Radix et Rhizoma Praeparata cum Melle,Rehmannine Radix,Ophiopogonis Radix,Astragali Radix and Codonopsis Radix.The clustering analysis results showed that the efficacy was mainly tonifying deficiency,regulating qi,activating blood circulation,and resolving stasis.Conclusion Professor Yuan Jinsheng's prescription compatibility for treating palpitations primarily focuses on tonics,qi-regulating,and blood circulation-activating and stasis-resolving herbs,embodying the principles of treating palpitations from the perspective of the heart and spleen and the combined use of multiple organs.

Objective: To explore the clinical dosimetry advantages of automated plan of IMRT for postoperative esophageal cancer and the dose prediction accuracy of the constructed 3D U-Res-Net model． Methods: A total of 110 postoperative esophageal cancer (middle and upper) cases treated by IMRT were considered in the study，of which 90 cases were randomly selected for training of deep learning prediction model．The deep learning prediction model and Auto-Plan module ( Philips pinnacle3 16. 2 ) were used to predict the three-dimension dose distribution and redesigned the remaining 20 cases respectively ，and the results obtained were compared with manual plan. Results : The average DSC value between the deep learning prediction plan and the manual plan was greater than 0. 92 in isodose surface，and the average Hausdorff distance HD95 of the isodose surface was 0. 58－0. 62 cm ; The V20 ，V30 ，Dmean of total lung were slightly lower than those of manual plan (P ＜0. 05 ) for the prediction model， meanwhile，the D2 ，D50 ，Dmean，HI of the target area and V30 of total lungs were better than those of manual plan(P ＜0. 05) for Auto-Plan ; Three-dimensional dose distribution of the three groups and the corresponding DVH curve showed that the three-dimensional dose distribution of the three groups had a little differences，and the DVH curves of the target area and organs at risk had a good agreement． Conclusion Auto-Plan can realize the design of automated plan for postoperative esophageal cancer，while the deep learning prediction model can realize the accurate prediction of the 3D dose distribution.

Hypertensive intracerebral hemorrhage (ICH) is mostly single in basal ganglia, thalamus and pons. Simultaneous hemorrhage in other brain regions is relatively rare, accounting for only 5.6% of all hemorrhagic strokes, while bilateral symmetrical hemorrhage is extremely rare. A case of bilateral basal ganglia symmetrical hemorrhage is reported for clinical reference.

Objective:To explore the efficacy and safety of intraarterial microguidewire electrocoagulation in arterial aneurysms.Methods:(1) SilverSpeed, a kind of microguidewire used in clinical intravascular treatment for intracranial aneurysms, was used to conduct in vitro electrolysis gas generation experiment with isolated arterial blood of anticoagulant New Zealand white rabbits as medium, and thrombus attachment on the surface of microguidewire was observed under scanning electron microscope. (2) Rabbit common carotid artery aneurysm models were established by using vein bag transplantation method, and divided into microguidewire electrocoagulation treatment groups ( n=40) and blank control group ( n=10). The number of closured tumor cavity and the quality of formed thrombus were observed after electrocoagulation simulation treatment with SilverSpeed microguidewire (charging at 6, 9, 12, 15, and 18 V voltage, respectively for 1, 3, 6, 9, 12, and 15 min). DSA was used to observe whether there was ruptured aneurysms or thrombosis of parent artery. Twelve h later, head MRI diffusion weighted sequence scan was performed to detect whether there were new cerebral ischemia foci in the distal cerebral blood supply area of the parent artery. DSA was performed again 6 months after surgery to observe whether the aneurysms recurred. Results:(1) Electrolytic gas generation experiment results showed that bubbles were generated after electrification of SilverSpeed microguidewire; the higher the voltage, the more severe the reaction. Scanning electron microscope showed that thrombus attached to the surface of the microguidewire after electrification in isolated blood; and the higher the voltage, the denser the thrombus. (2) Under the same charging time, the higher the voltage, the larger the number of closured tumor cavity in rabbits of the microguidewire electrocoagulation treatment groups. Under the same voltage, the longer the charging time, the better the quality of thrombosis. Ischemic events occurred only in the microguidewire electrocoagulation treatment group with voltage>9 V, and the charging duration was not associated with the incidence of embolic events. When the voltage was 15 V, 2 experimental rabbits died due to aneurysm rupture 3 min after electrification. When the voltage was 18 V, 4 experimental rabbits died of cardiac arrest 9 min after electrification, and another 2 rabbits died of aneurysm rupture 6 min after electrification.Conclusions:High voltage is the main cause of adverse events in the microguidewire electrocoagulation treatment of aneurysms. After setting the appropriate voltage, prolonging the electrification time can improve the electrocoagulation effect without increasing the safety risk.

Objective:To compare the dosimetric difference between intensity-modulated photon radiaotherapy (IMRT) planning and intensity-modulated proton radiotherapy (IMPT) planning for glioma.Methods:The clinical data of 15 glioma patients who underwent IMRT in ion medical center of the First Affiliated Hospital of USTC from November 2020 to April 2022 were retrospectively analyzed. IMRT planning and IMPT planning were designed for the image of each patient in the therapy planning system. Main dosimetric parameters were compared including plan target volume (PTV), coverage index (CI), dose homogeneity index (HI), and maximal dose (D max) and mean dose (D mean) of organs at risk between both plans. Results:There were no significant differences between IMRT planning and IMPT planning in terms of D max and D mean of PTV1 and PTV2, CI and HI (all P > 0.05). Compared with IMRT planning, brainstem D mean [6.92 GyE (0.09 GyE, 12.58 GyE) vs. 24.41 GyE (2.59 GyE, 34.18 GyE)], left optic nerve D max [0.78 GyE (0.04 GyE, 25.18 GyE) vs. 20.42 GyE (6.38 GyE, 37.17 GyE)], left optic nerve D mean [0.10 GyE (0.01 GyE, 11.63 GyE) vs. 9.74 GyE (2.99 GyE, 20.87 GyE)], right optic nerve D mean [1.57 GyE (0.13 GyE, 14.90 GyE) vs. 14.08 GyE (2.66 GyE, 23.67 GyE)], left len D max [0 GyE (0 GyE, 2.91 GyE) vs. 4.84 GyE (1.42 GyE, 5.48 GyE)], left len D mean [0 GyE (0 GyE, 1.73 GyE) vs. 3.84 GyE (1.25 GyE, 4.30 GyE)], right len D max [0.25 GyE (0.04 GyE, 4.55 GyE) vs. 4.28 GyE (1.58 GyE, 5.84 GyE)], right len D mean [0.16 GyE (0.01 GyE, 1.95 GyE) vs. 3.73 GyE (1.04 GyE, 4.86 GyE)], pituitary D max [6.97 GyE (0.18 GyE, 39.70 GyE) vs. 36.60 GyE (2.74 GyE, 45.19 GyE)], pituitary D mean [1.36 GyE (0.06 GyE, 13.85 GyE) vs. 24.74 GyE (2.42 GyE, 32.80 GyE)], hippocampus D max [5.10 GyE (0.24 GyE, 26.52 GyE) vs. 35.83 GyE (5.03 GyE, 46.11 GyE)], hippocampus D mean [0.36 GyE (0.04 GyE, 25.65 GyE) vs. 18.79 GyE (2.37 GyE, 28.10 GyE)] in IMPT planning were lower, and the differences were statistically significant (all P < 0.05). There were no statistical differences in brainstem D max [51.98 GyE (0.66 GyE, 53.43 GyE) vs. 53.29 GyE (3.87 GyE, 53.48 GyE)], right optic nerve D max [9.60 GyE (0.01 GyE, 43.32 GyE) vs. 25.37 GyE (3.45 GyE, 41.25 GyE)] of both plans (all P > 0.05). Conclusion:In the radiotherapy for glioma, IMRT and IMPT can meet the dose demand in clinic. Furthermore, IMPT planning can protect organs at risk and reduce radiation dose in hippocampus, brainstem, optic nerve, lens and pituitary.

Objective:To investigate the effects of ginsenoside Rg1 (GS-Rg1) on the secretion of exosomes (MSC-Exo) and expression of angiogenesis related microRNAs (miRNAs) in mesenchymal stem cells.Methods:Human umbilical cord blood mesenchymal stem cells (hUCBMSCs) were divided into experimental group and control group. The experimental group was treated with GS-RG1 at a final concentration of 40 mg/L, while the control group was treated with phosphate buffered saline (PBS) at the same volume. Both groups were cultured for 24 h. The morphology of MSC-Exo was observed by transmission electron microscopy; the characteristic surface markers were identified by Western blot; the concentration of MSC-Exo was detected by dicootanobutyric acid protein quantification method, and the expression of 8 miRNAs related to angiogenesis in MSC-Exo was detected by reverse transcription polymerase chain reaction (RT-PCR).Results:After 24 h of incubation, MSC-Exo with a circular membrane vesicle structure was visible. MSC-Exo was positive for the expression of the characteristic surface markers CD9, CD63 and TSG101. After 24 h of intervention, the concentration of MSC-Exo protein were (1.080±0.019)μg/μl and (0.881±0.032)μg/μl in the experimental group and control group, respectively, with statistically significant difference ( P<0.01). The expression of miR-126-3p, miR-21, miR-146a-5p and miR-125b-5p in the GS-Rg1 group were significantly higher than that in the control group, while the expression of miR-16-5p was significantly lower than that in the control group (all P<0.05). Conclusions:GS-Rg1 promotes the secretion of MSC-Exo and enhances the expression of angiogenesis-related miRNAs within Exo to promote angiogenesis.

The oral and maxillofacial region contains oral organs and facial soft tissues. Due to the complexity of the structures and functions of this region, the repair of related defects is complicated. Different degrees of defects require different repair methods, which involve a great combination of medicine and art, and the material requirements are extremely high. Hence, clinicians are plagued by contemporary oral repair materials due to the limitations of bone harvesting, immune rejection, low osteogenic activity and other problems. Decellularized extracellular matrix has attracted much attention as a bioactive scaffold material because of its nonimmunogenic properties, good osteogenic properties, slow release of growth factors, promotion of seed cell adhesion and maintenance of stem cell characteristics. This article reviews the sources, preparation methods, application and research progress of extracellular matrix materials in the repair of oral and maxillofacial defects to provide an overview for fundamental research and clinical development.