1.Preparation of Triptolide-Chuanxiong Rhizoma Extract Ethanol Transfersomes and Analysis on Its in Vitro Anti-inflammatory Mechanism
Ling TAO ; Zhiyan WAN ; Yidan LIU ; Zhe LI ; Zhenzhong ZANG ; Weifeng ZHU ; Yongmei GUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):210-218
ObjectiveTo prepare triptolide-Chuanxiong Rhizoma extract ethanol transfersomes(TP-CX@TESs), conduct its quality evaluation, and investigate its in vitro anti-inflammatory efficacy and the underlying mechanisms. MethodsTP-CX@TESs was prepared via the ultrasonic injection method. With encapsulation efficiency and particle size as evaluation indicators, Box-Behnken design-response surface methodology(BBD-RSM) was employed to optimize the formulation process. The TP-CX@TESs prepared under the optimal process was characterized and evaluated for in vitro transdermal performance. A lipopolysaccharide(LPS)-induced RAW264.7 cell inflammation model was established. After 24 h of drug intervention, the levels of inflammatory factors such as nitric oxide(NO), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in the cell supernatant were detected. Western blot was used to determine the protein expression levels of Janus kinase 2(JAK2), signal transducer and activator of transcription 3(STAT3), and α7 nicotinic acetylcholine receptor(α7nAChR), and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was applied to measure the mRNA expression levels of JAK2, STAT3, the encoding gene of α7nAChR(CHRNA7), and nuclear transcription factor-κB(NF-κB). ResultsResults of BBD-RSM showed that the optimal formulation for preparing TP-CX@TESs was as follows:egg yolk lecithin content of 2.3%, ethanol volume fraction of 30%, and ratio of polysorbate-80 to egg yolk lecithin of 2∶5. Microscopic characterization revealed that TP-CX@TESs exhibited a spherical-like structure with a particle size of (105.60±3.85) nm, a polydispersity index of 0.19±0.03, and a Zeta potential of (-15.89±0.98) mV. The encapsulation efficiencies of triptolide, ferulic acid, and ligustilide were (76.88±4.40)%, (78.84±4.40)%, and (65.88±0.06)%, respectively. Both in vitro release and transdermal penetration of triptolide, ferulic acid, and ligustilide in TP-CX@TESs all followed the first-order kinetic model, showing a certain sustained-release property. Experimental results in RAW264.7 cells indicated that TP-CX@TESs significantly inhibited the release of NO, TNF-α, and IL-6(P<0.01), remarkably upregulated the protein expression levels of STAT3 and α7nAChR(P<0.01), increased the mRNA expression level of CHRNA7, and significantly downregulated the mRNA expression level of NF-κB(P<0.05, P<0.01). ConclusionThe optimized formulation process of TP-CX@TESs is simple and feasible, along with favorable in vitro release property, good transdermal permeability, and excellent in vitro anti-inflammatory activity, the mechanism is related to the inhibition of NF-κB.
2.Preparation of Triptolide-Chuanxiong Rhizoma Extract Ethanol Transfersomes and Analysis on Its in Vitro Anti-inflammatory Mechanism
Ling TAO ; Zhiyan WAN ; Yidan LIU ; Zhe LI ; Zhenzhong ZANG ; Weifeng ZHU ; Yongmei GUAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):210-218
ObjectiveTo prepare triptolide-Chuanxiong Rhizoma extract ethanol transfersomes(TP-CX@TESs), conduct its quality evaluation, and investigate its in vitro anti-inflammatory efficacy and the underlying mechanisms. MethodsTP-CX@TESs was prepared via the ultrasonic injection method. With encapsulation efficiency and particle size as evaluation indicators, Box-Behnken design-response surface methodology(BBD-RSM) was employed to optimize the formulation process. The TP-CX@TESs prepared under the optimal process was characterized and evaluated for in vitro transdermal performance. A lipopolysaccharide(LPS)-induced RAW264.7 cell inflammation model was established. After 24 h of drug intervention, the levels of inflammatory factors such as nitric oxide(NO), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in the cell supernatant were detected. Western blot was used to determine the protein expression levels of Janus kinase 2(JAK2), signal transducer and activator of transcription 3(STAT3), and α7 nicotinic acetylcholine receptor(α7nAChR), and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was applied to measure the mRNA expression levels of JAK2, STAT3, the encoding gene of α7nAChR(CHRNA7), and nuclear transcription factor-κB(NF-κB). ResultsResults of BBD-RSM showed that the optimal formulation for preparing TP-CX@TESs was as follows:egg yolk lecithin content of 2.3%, ethanol volume fraction of 30%, and ratio of polysorbate-80 to egg yolk lecithin of 2∶5. Microscopic characterization revealed that TP-CX@TESs exhibited a spherical-like structure with a particle size of (105.60±3.85) nm, a polydispersity index of 0.19±0.03, and a Zeta potential of (-15.89±0.98) mV. The encapsulation efficiencies of triptolide, ferulic acid, and ligustilide were (76.88±4.40)%, (78.84±4.40)%, and (65.88±0.06)%, respectively. Both in vitro release and transdermal penetration of triptolide, ferulic acid, and ligustilide in TP-CX@TESs all followed the first-order kinetic model, showing a certain sustained-release property. Experimental results in RAW264.7 cells indicated that TP-CX@TESs significantly inhibited the release of NO, TNF-α, and IL-6(P<0.01), remarkably upregulated the protein expression levels of STAT3 and α7nAChR(P<0.01), increased the mRNA expression level of CHRNA7, and significantly downregulated the mRNA expression level of NF-κB(P<0.05, P<0.01). ConclusionThe optimized formulation process of TP-CX@TESs is simple and feasible, along with favorable in vitro release property, good transdermal permeability, and excellent in vitro anti-inflammatory activity, the mechanism is related to the inhibition of NF-κB.
3.Combination of Components from Tripterygii Radix et Rhizoma-Chuanxiong Rhizoma Affects RA-FLSs by Regulating NF-κB, Nrf2/HO-1 Signaling Pathways and Bcl-2/Caspase-3 Expression
Yongmei GUAN ; Zhiyan WAN ; Shuhui WANG ; Weifeng ZHU ; Zhiyong LIU ; Cheng JIANG ; Zhenzhong ZANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):17-26
ObjectiveTo investigate the effects of the combination of components from Tripterygii Radix et Rhizoma and Chuanxiong Rhizoma on rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) and the underlying mechanism. MethodsRA-FLSs were grouped as follows: blank control, positive control (methotrexate), Tripterygii Radix et Rhizoma components, Chuanxiong Rhizoma components, and components from Tripterygii Radix et Rhizoma+Chuanxiong Rhizoma. The cell-counting kit-8 (CCK-8) assay was employed to the cell proliferation, invasion, and apoptosis. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, reactive oxygen species (ROS), and malondiadehyde (MDA) in cells were measured. Western blot was employed to determine the protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), nuclear factor-kappa B (NF-κB) p65, phosphorylated inhibitory subunit of NF-κBα (p-IκBα), cysteinyl aspartate-specific protease-3 (Caspase-3), and B-cell lymphoma 2 (Bcl-2). Real-time PCR was employed to determine the mRNA levels of Nrf2, HO-1, and NF-κB p65. ResultsThe cells in the groups of positive control, Tripterygii Radix et Rhizoma components, Chuanxiong Rhizoma components, and components from Tripterygii Radix et Rhizoma+Chuanxiong Rhizoma were treated with 2.50 mg·L-1 methotrexate, 0.20 mg·L-1 triptolide + 0.20 mg·L-1 celastrol, 5.00 mg·L-1 ferulic acid + 20.00 mg·L-1 ligustrazine, 0.20 mg·L-1 triptolide + 0.20 mg·L-1 celastrol + 5.00 mg·L-1 ferulic acid + 20.00 mg·L-1 ligustrazine, respectively. Compared with the blank control group, drug administration reduced the proliferation and invasion and increased the apoptosis of cells (P<0.01), lowered the levels of TNF-α, IL-6, ROS, and MDA (P<0.01), up-regulated the mRNA and protein levels of Caspase-3, Nrf2, and HO-1 (P<0.01), and down-regulated the mRNA and protein levels of Bcl-2, NF-κB p65, and p-IκBα (P<0.01). Compared with the Tripterygii Radix et Rhizoma components group, the combination of components from Tripterygii Radix et Rhizoma+Chuanxiong Rhizoma inhibited the proliferation and invasion (P<0.05) and promoted the apoptosis of RA-FLSs, up-regulated the mRNA levels of Nrf2 and HO-1 and protein levels of Nrf2 and Caspase-3 (P<0.05), and down-regulated the protein levels of NF-κB p65 and p-IκBα (P<0.05). ConclusionThe combination of components from Chuanxiong Rhizoma and Tripterygii Radix et Rhizoma can inhibit the proliferation and invasion and promote the apoptosis of RA-FLSs and alleviate oxidative stress and inflammation by inhibiting the NF-κB signaling pathway, activating the Nrf2/HO-1 pathway, and regulating the expression of Bcl-2/Caspase-3.
4.Analysis of the diagnostic value of mp-MRI combined with CT for brucellar spondylitis
Yueling REN ; Jingshu HU ; Batekxike TUOERGEN ; Jianhong FANG ; Hongrui ZHOU ; Weifeng GUAN
Chinese Journal of Endemiology 2025;44(5):415-419
Objective:To explore the diagnostic value of multiparametric magnetic resonance imaging (mp-MRI) combined with computed tomography (CT) for brucellar spondylitis (BS).Methods:From March 2019 to March 2024, patients with suspected BS admitted to Yili Kazakh Autonomous Prefecture Friendship Hospital were selected as study subjects. Their clinical characteristics, laboratory tests, and imaging findings (mp-MRI and CT) were analyzed.Results:A total of 60 suspected BS patients were enrolled, including 43 males and 17 females. Among them, 39 cases (65.00%) were diagnosed with BS, while 21 cases (35.00%) were non-BS. CT examination was performed in 48 patients, mp-MRI in 40 patients, and both examinations in 28 patients. Among CT-examined patients, there were 28 BS cases, 1 case had cervical spine involvement, 3 case had thoracic spine involvement, 3 case had thoracolumbar spine involvement, and 21 case had lumbosacral spine involvement. There were 16 cases involved 2 vertebrae, 3 cases involved 3 vertebrae, and 9 cases involved > 3 vertebrae.There were 15 cases had vertebral osteophyte formation, 28 cases had vertebral bone destruction, 22 cases had intervertebral space stenosis, 25 cases had uneven intervertebral disc density, 28 cases had thickening of soft tissues around the vertebrae, 19 cases had paravertebral abscesses, and 6 cases had spinal canal stenosis. In the mp-MRI examination of patients, there were 30 BS patients, among whom 1 case had cervical spine involvement, 2 case had thoracic spine involvement, 4 case had thoracolumbar spine involvement, and 23 case had lumbosacral spine involvement. There were 16 cases involved 2 vertebrae, 3 cases involved 3 vertebrae, and 11 cases involved > 3 vertebrae, 24 cases had intervertebral space stenosis, 27 cases had intervertebral disc destruction. There were 27 cases had swelling and thickening of soft tissues around the vertebrae, and 21 cases had paravertebral, psoas major muscle, and/or spinal canal abscesses. The BS group showed lower fractional anisotropy value and higher apparent diffusion coefficient value compared to the non-BS group, with statistically significant differences ( t = 5.26, 5.63, P < 0.001). The mp-MRI combined with CT showed higher sensitivity (86.36%, 19/22) and specificity (5/6) for BS diagnosis compared to mp-MRI alone (78.57%, 11/14; 80.77%, 21/26) and CT alone (56.25%, 9/16; 71.88%, 23/32). Conclusions:BS patients present diverse imaging manifestations. The combination of mp-MRI and CT demonstrates better diagnostic efficacy for BS than single examinations alone, showing good diagnostic value.
5.Effect of preoperative anxiety on consciousness and autonomic nervous activity during propofol anesthesia
Guanqi MA ; Ying HU ; Weifeng TU ; Cui LI ; Jinping GUAN ; Junlong ZHANG
Chinese Journal of Anesthesiology 2025;45(1):42-48
Objective:To evaluate the effect of preoperative anxiety on the consciousness and autonomic nervous activity during propofol anesthesia.Methods:This study was a secondary analysis of data from the clinical trial in a prospective single-arm study. One hundred and thirty patients, aged 18-65 yr, with a body mass index of 18.5-27.9 kg/m 2, of American Society of Anesthesiologists Physical Status classification I or Ⅱ, scheduled to receive propofol anesthesia, were selected from the Second People′s Hospital of Lianyungang. The six-item of the state anxiety inventory (SAI) of the State-Trait Anxiety Inventory was used to assess the anxiety of patients 1 h before surgery. The patients were divided into 2 groups according to the cut-off value of 12: obvious anxiety symptom (SAI score >12) group (group A, n=49) and no obvious anxiety symptom (SAI score ≤12) group (group B, n=81). After admission to the operating room, the patient was required to hold a 50 ml syringe filled with water. Propofol was given by target-controlled infusion (TCI) with the target plasma concentration set at 5 μg/ml. When the effect-site concentration (Ce) of propofol increased to 3.5 μg/ml (all the patients lost consciousness), the closed-loop TCI was used to maintain BIS value between 45 and 55. The patients were monitored for 20 min after stopping the pump infusion (anesthesia recovery period). The disappearance time of verbal command, disappearance time of eyelash reflex, time of syringe dropping, recovery time of verbal command, recovery time of eyelash reflex, Ce at the recovery of verbal command, Ce at the recovery of eyelash reflex, Ce within the first 5 min of the closed-loop TCI, and consumption of propofol during anesthesia were recorded. The peripheral perfusion index, low frequency power and high frequency power of heart rate variability were recorded, and the ratio of low frequency power to high frequency power was calculated. Pearson correlation analysis was used to assess the correlation between preoperative SAI score and consciousness-related indicators, simulated Ce of propofol and consumption of propofol. Results:Compared with group B, the disappearance time of verbal command, disappearance time of eyelash reflex, and time of syringe dropping were significantly prolonged, the consumption of propofol, simulated Ce at recovery of verbal command and within the first 5 min of closed-loop TCI were increased, the peripheral perfusion index was decreased at each time point before administration and at 14-20 min of anesthesia recovery, and the low-frequency power was decreased during anesthesia maintenance in group A ( P<0.05). The SAI score was positively correlated with the disappearance time of verbal command ( r=0.220, P=0.012), time of syringe dropping ( r=0.206, P=0.029), consumption of propofol ( r=0.330, P<0.001), and the simulated Ce at the recovery of verbal command ( r=0.215, P=0.015) and simulated Ce at recovery of eyelash reflex ( r=0.207, P=0.022). Conclusions:Preoperative anxiety may lead to prolonged loss of consciousness and more marked inhibition of sympathetic nerve activity during propofol anesthesia.
6.Effect of preoperative anxiety on consciousness and autonomic nervous activity during propofol anesthesia
Guanqi MA ; Ying HU ; Weifeng TU ; Cui LI ; Jinping GUAN ; Junlong ZHANG
Chinese Journal of Anesthesiology 2025;45(1):42-48
Objective:To evaluate the effect of preoperative anxiety on the consciousness and autonomic nervous activity during propofol anesthesia.Methods:This study was a secondary analysis of data from the clinical trial in a prospective single-arm study. One hundred and thirty patients, aged 18-65 yr, with a body mass index of 18.5-27.9 kg/m 2, of American Society of Anesthesiologists Physical Status classification I or Ⅱ, scheduled to receive propofol anesthesia, were selected from the Second People′s Hospital of Lianyungang. The six-item of the state anxiety inventory (SAI) of the State-Trait Anxiety Inventory was used to assess the anxiety of patients 1 h before surgery. The patients were divided into 2 groups according to the cut-off value of 12: obvious anxiety symptom (SAI score >12) group (group A, n=49) and no obvious anxiety symptom (SAI score ≤12) group (group B, n=81). After admission to the operating room, the patient was required to hold a 50 ml syringe filled with water. Propofol was given by target-controlled infusion (TCI) with the target plasma concentration set at 5 μg/ml. When the effect-site concentration (Ce) of propofol increased to 3.5 μg/ml (all the patients lost consciousness), the closed-loop TCI was used to maintain BIS value between 45 and 55. The patients were monitored for 20 min after stopping the pump infusion (anesthesia recovery period). The disappearance time of verbal command, disappearance time of eyelash reflex, time of syringe dropping, recovery time of verbal command, recovery time of eyelash reflex, Ce at the recovery of verbal command, Ce at the recovery of eyelash reflex, Ce within the first 5 min of the closed-loop TCI, and consumption of propofol during anesthesia were recorded. The peripheral perfusion index, low frequency power and high frequency power of heart rate variability were recorded, and the ratio of low frequency power to high frequency power was calculated. Pearson correlation analysis was used to assess the correlation between preoperative SAI score and consciousness-related indicators, simulated Ce of propofol and consumption of propofol. Results:Compared with group B, the disappearance time of verbal command, disappearance time of eyelash reflex, and time of syringe dropping were significantly prolonged, the consumption of propofol, simulated Ce at recovery of verbal command and within the first 5 min of closed-loop TCI were increased, the peripheral perfusion index was decreased at each time point before administration and at 14-20 min of anesthesia recovery, and the low-frequency power was decreased during anesthesia maintenance in group A ( P<0.05). The SAI score was positively correlated with the disappearance time of verbal command ( r=0.220, P=0.012), time of syringe dropping ( r=0.206, P=0.029), consumption of propofol ( r=0.330, P<0.001), and the simulated Ce at the recovery of verbal command ( r=0.215, P=0.015) and simulated Ce at recovery of eyelash reflex ( r=0.207, P=0.022). Conclusions:Preoperative anxiety may lead to prolonged loss of consciousness and more marked inhibition of sympathetic nerve activity during propofol anesthesia.
7.Analysis of the diagnostic value of mp-MRI combined with CT for brucellar spondylitis
Yueling REN ; Jingshu HU ; Batekxike TUOERGEN ; Jianhong FANG ; Hongrui ZHOU ; Weifeng GUAN
Chinese Journal of Endemiology 2025;44(5):415-419
Objective:To explore the diagnostic value of multiparametric magnetic resonance imaging (mp-MRI) combined with computed tomography (CT) for brucellar spondylitis (BS).Methods:From March 2019 to March 2024, patients with suspected BS admitted to Yili Kazakh Autonomous Prefecture Friendship Hospital were selected as study subjects. Their clinical characteristics, laboratory tests, and imaging findings (mp-MRI and CT) were analyzed.Results:A total of 60 suspected BS patients were enrolled, including 43 males and 17 females. Among them, 39 cases (65.00%) were diagnosed with BS, while 21 cases (35.00%) were non-BS. CT examination was performed in 48 patients, mp-MRI in 40 patients, and both examinations in 28 patients. Among CT-examined patients, there were 28 BS cases, 1 case had cervical spine involvement, 3 case had thoracic spine involvement, 3 case had thoracolumbar spine involvement, and 21 case had lumbosacral spine involvement. There were 16 cases involved 2 vertebrae, 3 cases involved 3 vertebrae, and 9 cases involved > 3 vertebrae.There were 15 cases had vertebral osteophyte formation, 28 cases had vertebral bone destruction, 22 cases had intervertebral space stenosis, 25 cases had uneven intervertebral disc density, 28 cases had thickening of soft tissues around the vertebrae, 19 cases had paravertebral abscesses, and 6 cases had spinal canal stenosis. In the mp-MRI examination of patients, there were 30 BS patients, among whom 1 case had cervical spine involvement, 2 case had thoracic spine involvement, 4 case had thoracolumbar spine involvement, and 23 case had lumbosacral spine involvement. There were 16 cases involved 2 vertebrae, 3 cases involved 3 vertebrae, and 11 cases involved > 3 vertebrae, 24 cases had intervertebral space stenosis, 27 cases had intervertebral disc destruction. There were 27 cases had swelling and thickening of soft tissues around the vertebrae, and 21 cases had paravertebral, psoas major muscle, and/or spinal canal abscesses. The BS group showed lower fractional anisotropy value and higher apparent diffusion coefficient value compared to the non-BS group, with statistically significant differences ( t = 5.26, 5.63, P < 0.001). The mp-MRI combined with CT showed higher sensitivity (86.36%, 19/22) and specificity (5/6) for BS diagnosis compared to mp-MRI alone (78.57%, 11/14; 80.77%, 21/26) and CT alone (56.25%, 9/16; 71.88%, 23/32). Conclusions:BS patients present diverse imaging manifestations. The combination of mp-MRI and CT demonstrates better diagnostic efficacy for BS than single examinations alone, showing good diagnostic value.
8.Application and significance of prone position in the treatment of patients with severe pneumonia in intensive care unit
Huiyan YU ; Chun GUAN ; Weifeng XIE ; Qingshu LI ; Yan QU ; Yu LUO ; Dan HU
Chinese Critical Care Medicine 2024;36(4):364-368
Objective:To investigate the effect of prone position on the prognosis of patients with severe pneumonia in intensive care unit (ICU).Methods:A retrospective cohort study was conducted. The patients with severe pneumonia admitted to the ICU of Qingdao Municipal Hospital from May 2022 to August 2023 were enrolled. The general information, etiology, underlying diseases, vital signs and laboratory indicators at ICU admission, clinical treatment and prognosis during ICU hospitalization were collected. The above clinical data of patients with different prognosis were compared. Multifactorial Logistic regression analysis was used to screen the related factors affecting survival during ICU in patients with severe pneumonia. The change in oxygenation index (PaO 2/FiO 2) of patients with severe pneumonia were observed at 1 hour before the first prone position, 1 hour after the first prone position, and 1 hour after the end of the first prone position. The effect of prone position on oxygenation in patients with severe pneumonia was analyzed. Spearman correlation analysis was used to investigate the correlation between the duration to first prone position and the change in the PaO 2/FiO 2 before and after prone position in patients with severe pneumonia. Results:Finally, a total of 144 patients with severe pneumonia were enrolled, 45 survived and 99 died during ICU hospitalization, with a mortality of 68.8%. Compared with the survival group, the patients in the death group were older [years old: 81.00 (70.75, 86.00) vs. 71.00 (60.50, 81.50), P < 0.01], the proportion of pre-existing lung disease, heart rate (HR), respiratory rate (RR), blood lactic acid (Lac) and the ratio of continuous renal replacement therapy (CRRT) were higher [ratio of pre-existing lung disease: 23.2% (23/99) vs. 8.9% (4/45), HR (bpm): 99.61±22.47 vs. 91.49±18.76, RR (times/min): 22.50 (19.75, 29.25) vs. 20.00 (17.50, 24.50), Lac (mmol/L): 2.00 (1.55, 3.25) vs. 1.60 (1.20, 1.95), CRRT ratio: 25.3% (25/99) vs. 6.7% (3/45), all P < 0.05], and the proportion of prone position was lower [41.4% (41/99) vs. 68.9% (31/45), P < 0.01]. Multifactorial Logistic regression analysis showed that age [odds ratio ( OR) = 0.946, 95% confidence interval (95% CI) was 0.912-0.980, P = 0.002] and Lac ( OR = 0.563, 95% CI was 0.340-0.930, P = 0.025) were negatively correlated with survival during ICU hospitalization in severe pneumonia patients, while prone position was positively correlated with survival ( OR = 2.551, 95% CI was 1.067-6.095, P = 0.035), indicating that prone position was beneficial for improving ICU prognosis in severe pneumonia patients. The results of PaO 2/FiO 2 at different time points in prone position showed that PaO 2/FiO 2 at 1 hour of the first prone position in the patients with severe pneumonia was significantly higher than that at 1 hour before the first prone position [mmHg (1 mmHg ≈ 0.133 kPa): 146.69 (113.92, 257.25) vs. 111.75 (70.15, 212.20), P < 0.01], indicating that the prone position had a relevant effect on the improvement of oxygenation in patients. Spearman correlation analysis showed that the duration of the first prone position in patients with severe pneumonia was significantly and positively correlated with the improvement of oxygenation at 1 hour of the first prone position ( r = 0.565, P < 0.001). Conclusions:The prone position is a therapeutic measure that can independently influence the prognosis of patients with severe pneumonia during ICU hospitalization. The prone position effectively improves oxygenation in patients with severe pneumonia and the first change in oxygenation in patients is related to the duration of the prone position.
9.Effect and mechanism of human adipose-derived stem cell exosomes on diabetic peripheral neuropathy
Tao CAO ; Tong HAO ; Dan XIAO ; Weifeng ZHANG ; Peng JI ; Yanhui JIA ; Jing WANG ; Xujie WANG ; Hao GUAN ; Ke TAO
Chinese Journal of Burns 2024;40(3):240-248
Objective:To investigate the changes of artemin protein expression in diabetic peripheral neuropathy (DPN) and to explore the regulatory effect of human adipose-derived stem cell (ADSC) exosomes on the change of artemin protein expression.Methods:This research was a prospective observational clinical research combined with experimental research. Thirteen DPN patients (9 males and 4 females, aged 32 to 68 years) who were admitted to the First Affiliated Hospital of Air Force Medical University (hereinafter referred to as our hospital) from May 2022 to October 2023 and met the inclusion criteria were selected as DPN group, and 5 non-diabetes patients (4 males and 1 female, aged 29 to 61 years) who were admitted to our hospital in the same period of time and met the inclusion criteria were selected as control group. The toe nerve or sural nerve tissue in the abandoned tissue after debridement or amputation of patients in the two groups was collected. The pathological changes of nerve tissue were observed after hematoxylin-eosin staining; the protein expressions of S100β and artemin in nerve tissue were observed after immunofluorescence staining, and the artemin protein expression was quantified; the protein and mRNA expressions of artemin were detected by Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction, respectively (the sample number in DPN group and control group was 13 and 5, respectively). Twelve male C57BL/6 mice aged 3 to 5 days were collected to isolate Schwann cells, and the cells were divided into conventional culture group cultured routinely, high glucose alone group (cultured with high concentration of glucose solution only), and high glucose+exosome group (cultured with high concentration of glucose solution and extracted human ADSC exosomes). After 24 hours of culture, the cell proliferation activity was detected by cell counting kit 8 ( n=6). After 48 hours of culture, the protein expression of artemin was detected by Western blotting ( n=3). Results:Compared with those in control group, the neural supporting cells decreased and the inflammatory cells increased in the nerve tissue of patients in DPN group, showing typical manifestations of nerve injury. Immunofluorescence staining showed that compared with those in control group, the nuclei was more, and the protein expression of S100β was lower in nerve tissue of patients in DPN group. The protein expression of artemin in nerve tissue of patients in DPN group was 71±31, which was significantly lower than 1 729±62 in control group ( t=76.92, P<0.05). Western blotting detection showed that the protein expression of artemin in nerve tissue of patients in DPN group was 0.74±0.08, which was significantly lower than 0.97±0.06 in control group ( t=5.49, P<0.05). The artemin mRNA expression in nerve tissue of patients in DPN group was significantly lower than that in control group ( t=7.65, P<0.05). After 24 hours of culture, compared with that in conventional culture group, the proliferation activities of Schwann cells in high glucose alone group and high glucose+exosome group were significantly decreased ( P<0.05); compared with that in high glucose alone group, the proliferation activity of Schwann cells in high glucose+exosome group was significantly increased ( P<0.05). After 48 hours of culture, compared with those in conventional culture group, the protein expressions of artemin of Schwann cells in high glucose alone group and high glucose+exosome group were significantly decreased ( P<0.05); compared with that in high glucose alone group, the protein expression of artemin of Schwann cells in high glucose+exosome group was significantly increased ( P<0.05). Conclusions:The protein expression of artemin in nerve tissue of DPN patients is lower than that in normal nerve tissue, which may be related to the reduction of proliferation activity of Schwann cells by high glucose. Human ADSC exosomes may improve the proliferation activity of Schwann cells by increasing artemin protein expression, thereby delaying the progression of DPN.
10.Effect of volatile oil of Ligusticum chuanxiong on transdermal properties and cytotoxicity of triptolide in vitro
Cheng JIANG ; Zhenzhong ZANG ; Lihua CHEN ; Haiting ZHU ; Shihua FU ; Weifeng ZHU ; Wenting WU ; Wei SHI ; Yongmei GUAN
China Pharmacy 2024;35(4):413-418
OBJECTIVE To explore the effect of volatile oil of Ligusticum chuanxiong on the transdermal properties and cytotoxicity of triptolide in vitro. METHODS The chemical constituents of the volatile oil of L. chuanxiong were analyzed by gas chromatography-mass spectrometry. The lower abdominal skin of KM mice was separated and divided into triptolide group, triptolide in compatibility with volatile oil of L. chuanxiong groups at 1∶10, 1∶50, 1∶100 (hereinafter referred to as “compatibility 1∶10”“compatibility 1∶50”“compatibility 1∶100” groups). After the skin of mice in each group was fully exposed to 0.2 g of the corresponding cream for 24 h, the cumulative transdermal dose (Qn) of triptolide in the receiving solution of each group was determined by high-performance liquid chromatography, and the transdermal absorption rate (Jss) was calculated. Human immortalized keratinocytes (HaCat) were used as a model, the CCK-8 method was used to detect the cell survival rate of different concentrations of the volatile oil of L. chuanxiong and triptolide before and after compatibility. RESULTS A total of 62 chemical constituents of the volatile oil of L. chuanxiong were identified, including Z-ligustilide, senkyunolide, and β-selinene. The Qn (P< 0.01) and Jss of triptolide increased within 24 h in the compatibility 1∶10 and 1∶50 groups, while the Qn (P<0.05) and Jss decreased in the compatibility 1∶100 group as compared with the triptolide group. Compared with the triptolide group, the cell survival rate of HaCat was significantly increased in the compatibility 1∶10 and 1∶50 groups when the triptolide concentrations were 36, 72 and 144 ng/mL (P<0.05 or P<0.01); while the cell survival rate of HaCat was decreased in the compatibility 1∶100 group, but the difference was not statistically significant (P>0.05). CONCLUSIONS When the compatibility ratio of triptolide and volatile oil of L. chuanxiong was 1∶10 or 1∶50, it can promote the transdermal absorption of triptolide and reduce the cytotoxicity of triptolide to HaCat.

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