1.Mechanism of Wenfei Huaxian Decoction-containing Serum in Delaying Inflammatory Senescence of Lung Mesenchymal Stem Cells Based on NAMPT/SIRT1
Junxia HU ; Yueqi XU ; Jun WANG ; Guoshaung ZHU ; Shiwen KE ; Mingliang QIU ; Liangji LIU ; Lisha MO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(12):45-53
ObjectiveThe lung mesenchymal stem cells (LMSCs) induced by D-galactose (D-gal) were intervened by Wenfei Huaxian decoction-containing serum to explore the mechanism of Wenfei Huaxian decoction in delaying the senescence of LMSCs through the nicotinamide phosphoribosyltransferase/silent information regulator 1 (NAMPT/SIRT1) signaling pathway. MethodWenfei Huaxian decoction-containing serum was prepared. LMSCs were isolated by gradient density centrifugation, and they were cultured and identified in vitro. The senescence model in vitro was established by stimulating cells via D-gal for 24 h. LMSCs cells were modeled after being treated with different volume fractions (5%, 10%, 20%, 40%, and 80%) of Wenfei Huaxian decoction-containing serum for 24 h, and the cell proliferation level was detected by methyl thiazolyl tetrazolium (MTT) method. The cells were randomly divided into blank serum group, model group, and high, medium, and low dose groups of Wenfei Huaxian decoction-containing serum. Senescence-associated β-galactosidase (SA-β-gal) staining was used to detect the senescence of LMSCs in each group. The content of NAD + was detected by colorimetry. The levels of senescence-associated factors (p16 and p53), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in cell culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the relative expression of senescence-associated proteins and NAMPT/SIRT1 signaling pathway-related proteins. ResultCompared with the blank serum group, the proliferation of LMSCs was significantly inhibited after D-gal stimulation for 24 h (P<0.01). Compared with the model group, the proliferation of LMSCs could be promoted after intervention with the corresponding Wenfei Huaxian decoction-containing serum (P<0.05, P<0.01). Compared with the blank serum group, the SA-β-gal staining of LMSCs in the model group after D-gal stimulation was enhanced, and the content of NAD+ was increased (P<0.01). The expression levels of senescence factors p16 and p53, as well as SASP pro-inflammatory factors IL-6 and TNF-α in the cell culture supernatant, were significantly increased (P<0.01). The expression of senescence-associated proteins p16, p21, and p53 increased (P<0.01), and the protein expression of NAMPT, SIRT1, peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), and forkhead box family transcription factor O1 (FoxO1) decreased (P<0.01). Compared with the model group, the SA-β-gal staining of LMSCs in each group of Wenfei Huaxian decoction-containing serum was significantly reduced, and the content of NAD+ was decreased (P<0.01). The senescence factors (p16 and p53) and inflammatory factors (IL-6 and TNF-α) in the cell culture supernatant were significantly decreased (P<0.01). The expression of senescence-associated proteins (P16, P21, and P53) decreased (P<0.05, P<0.01). The protein expressions of NAMPT, SIRT1, PGC-1α, and FoxO1 were significantly up-regulated (P<0.05, P<0.01). ConclusionWenfei Huaxian decoction can alleviate senescence and inflammatory response damage of D-gal-induced LMSCs, and its mechanism may be related to the regulation of the NAMPT/SIRT1 signaling pathway.
2.Prediction of target genes and establishment of related prognostic model for the treatment of glioblastoma with stigmasterol
Qiang ZHU ; Ruichun LI ; Shiwen GUO ; Chen LIANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(6):909-917
[Objective] To predict potential target genes for the treatment of glioblastoma (GBM) with stigmasterol and construct a relevant prognostic model, in order to reveal its antiglioma mechanism and the role of these target genes in the prognosis of GBM patients. [Methods] Differential expression genes in GBM and stigmasterol target genes were obtained via online databases. Venn diagram was used to select potential target genes for stigmasterol treatment of GBM, and enrichment analysis was performed using R language. Univariate COX regression analysis and least absolute shrinkage and selection operator (LASSO) analysis were made to select stigmasterol target genes related to the prognosis of GBM patients and construct a relevant prognostic model. Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blotting analyses were used to detect the effect of stigmasterol on the expressions of related target genes. [Results] In this study, a total of 31 potential target genes for the treatment of GBM with stigmasterol were identified. Enrichment analysis showed that these target genes were associated with the activation of the G protein coupled receptor (GPCR) signaling pathway and the regulation of lipid metabolism. Regression analysis identified two stigmasterol target genes, namely, fatty acid binding protein 5 (FABP5) and alpha 1B adrenergic receptor (ADRA1B), which are associated with the prognosis of GBM. A prognostic model constructed based on these two genes could accurately predict the prognosis of GBM patients. Finally, stigmasterol inhibited the expressions of these two genes in GBM cells (FABP5: t=9.909, P=0.001; ADRA1B: t=3.319, P=0.029). [Conclusion] Stigmasterol’s anti-tumor effect may be linked to its regulation of GPCR signaling pathways and lipid metabolism. By inhibiting the expressions of FABP5 and ADRA1B, stigmasterol could potentially enhance the prognosis for GBM patients. Additionally, a prognostic model based on the expression levels of FABP5 and ADRA1B can be valuable for predicting patient outcomes and monitoring therapeutic efficacy in GBM.
3.The Effects of Wenfei Huaxian Decoction (温肺化纤汤) on Pulmonary Fibrosis and Endoplasmic Reticulum Stress in Systemic Sclerosis-Associated Interstitial Lung Disease Model Mice
Mingliang QIU ; Jiali XIONG ; Chenxiao XIAO ; Xinzhu ZHOU ; Lisha MO ; Shiwen KE ; Guoshuang ZHU ; Liangji LIU
Journal of Traditional Chinese Medicine 2024;65(13):1383-1391
ObjectiveTo investigate the possible mechanism of Wenfei Huaxian Decoction (温肺化纤汤) in treatment of pulmonary fibrosis in systemic sclerosis-associated interstitial lung disease (SSc-ILD). MethodsSixty C3H/He female rats were randomly divided into a control group, a model group, a pirfenidone group, and low-, medium-, and high-dose Wenfei Huaxian Decoction groups. The SSc-ILD model mice was established by subcutaneous injection of bleomycin solution 0.04 mg/d into the back of mice for 28 days in all groups but the control group. After successful modelling, the pirfenidone group was given pirfenidone capsule 300 mg/(kg·d) by gavage, the low-, medium- and high-dose Wenfei Huaxian Decoction groups were given Wenfei Huaxian Decoction 7.81, 15.62, and 31.24 g/(kg·d) by gavage, respectively, and the control group as well as the model group were given normal saline 0.1 ml/10 g by gavage, for a total of 21 days. At the end of the intervention, HE staining and Masson staining were used to observe the pathological changes in the skin and lung tissues; the hydroxyproline content of the skin and lung tissues was detected; the protein expression levels of endoplasmic reticulum stress-related proteins glucose-regulated protein 78 (BIP) and C/EBP homologous protein (CHOP) as well as those of nuclear factor kappa B (NF-κB) pathway p65 were measured by western blot; ELISA was performed to determine the expression levels of interferon gamma (IFN-γ), interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF-α) in serum of rats. ResultsThe results of HE and Masson staining indicated that compared with the control group, the dermis significantly thickened, the number of collagen fibers significantly enlarged, and the number of inflammatory cells significantly increased in the model group; the lung tissue showed a marked inflammatory cellular response with massive collagen fibre proliferation with inflammatory cell infiltration. Compared with the model group, the skin tissue and lung tissue collagen fibre proliferation significantly reduced and inflammatory cell infiltration reduced in the pirfenidone group and all dose groups of Wenfei Huaxian Decoction, and the effects of pirfenidone group and Wenfei Huaxian Decoction medium- and high-dose groups were basically comparable. Compared with the model group, the content of hydroxyproline in skin and lung tissue, the serum level of IFN-γ, IL-6 and TNF-α, and the expression levels of BIP and CHOP protein in lung tissue increased in model group (P<0.05). Compared with model group, the content of hydroxyproline in skin tissue of pirfenidone group, low-and medium-dose Wenfei Huaxian Decoction groups decreased, and the content of hydroxyproline in lung tissue of medium-dose Wenfei Huaxian Decoction group decreased. The serum level of IFN-γ, IL-6, TNF-α and the expression levels of BIP, CHOP and p65 protein in lung tissue of rats in pirfenidone group and high-dose Wenfei Huaxian Decoction group decreased (P<0.05). The content of hydroxyproline in lung tissue of medium-dose Wenfei Huaxian Decoction group was significantly lower than that of low-dose and high-dose Wenfei Huaxian Decoction group, and the serum level of IFN-γ, IL-6, TNF-α in low- and medium-dose Wenfei Huaxian Decoction group were higher than those in high-dose Wenfei Huaxian Decoction group. The expression level of BIP protein in high-dose group was significantly lower than that in low- and medium-dose Wenfei Huaxian Decoction groups (P<0.05). ConclusionWenfei Huaxian Decoction can improve the skin and lung fibrosis of SSc-ILD rats, which may act through anti-inflammation, inhibition of NF-κB pathway, and then inhibition of endoplasmic reticulum stress, which ultimately blocked the fibrotic process.
4.Correlation between CT imaging-derived hip muscle factors and recovery of independent mobility within 1 year after surgery in older adults with hip fractures
Yufeng GE ; Feng GAO ; Chao TU ; Ling WANG ; Gang LIU ; Wenshuang ZHANG ; Shiwen ZHU ; Minghui YANG ; Xinbao WU
Chinese Journal of Trauma 2024;40(6):531-538
Objective:To explore the correlation between hip muscle factors measured with CT imaging and recovery of independent mobility within 1 year after surgery in elderly patients with hip fractures.Methods:A prospective cohort study was conducted on the clinical data of 680 elderly patients with hip fractures admitted to Beijing Jishuitan Hospital of Capital Medical University from November 2018 to December 2019. The patients were assigned to dependent group and assistant group according to whether they regained pre-injury independent mobility within 1 year after surgery. Gender, age, body mass index, personal history, living habits, past diseases, Charlson comorbidity index, laboratory test indicators, fracture types, anesthesia types, surgical methods, rehabilitation training, time from injury to surgery, and hip muscle parameters in both groups were recorded. OsiriX software was employed in the measurement of the hip muscles to measure the muscle area and density of the gluteus maximus and gluteus medius/minimus on CT images, and the average values were calculated as hip muscle area and density. Then the variables of hip muscle area and density were converted seperately to gender-normalized Z-scores, and were divided into high-area group ( Z≥0) and low-area group ( Z<0), and high-density group ( Z≥0) and low-density group ( Z<0) respectively. Observable variables were primarily analyzed using univariate analysis between the independent group and assistant group. Those variables with statistically significant differences in the univariate analysis or would potentially affect mobility recovery according to previous researches although there were no statistical significance were included in a multivariate Logistic regression analysis. Three Logistic regression models were designed (Model 1 uncorrected, Model 2 corrected for gender, age and body mass index, Model 3 corrected for variables in Model 2 and other variables included after above-mentioned analysis) to analyze whether muscle parameters were risk factors for recovery of independent mobility. Additionally, generalized estimating equations were used for repeated measurement to analyze the correlation between hip muscle area and recovery of independent mobility after surgery. Results:Compared to the assistant group, the independent group were younger in age, with lower rate of living alone, being housebound, cognitive impairment, and Charlson comorbidity index, lower level of hemoglobin and albumin, higher rate of femoral neck fractures, lower rate of internal fixation, shorter time from injury to surgery, larger hip muscle area, and higher hip muscle density ( P<0.05 or 0.01). Multivariate Logistic regression analysis showed that, in the fully corrected Model 3, only hip muscle area remained significantly correlated with recovery of independent mobility ( P<0.05), while no significant difference was found between the high-density group and low-density group ( P>0.05). In the repeated measurement, patients in the high-area group were 1.84 times more likely to restore independent mobility than those in the low-area group ( OR=1.84, 95% CI 1.33, 2.53, P<0.01). Conclusions:Hip muscle area measured with CT imaging is closely correlated to the recovery of independent mobility within 1 year after surgery in elderly patients with hip fractures. Moreover, larger hip muscle area indicates a larger likelihood of recovery of independent mobility.
5.A digital classification system of pelvic fractures based on close reduction techniques
Xu SUN ; Yuneng LI ; Qiyong CAO ; Chunpeng ZHAO ; Yimin CHEN ; Minghui YANG ; Shiwen ZHU ; Honghua WU ; Xinbao WU
Chinese Journal of Orthopaedic Trauma 2024;26(5):428-434
Objective:To explore the feasibility and consistency of a new digital classification system of pelvic fractures named as JST classification based on close reduction techniques.Methods:A retrospective collection was conducted of the data from the 63 patients with pelvic fracture who had undergone surgical treatment after JST classification at Department of Orthopaedics and Traumatology, Beijing Jishuitan Hospital from March 2021 to March 2023. Digital classification of the pelvic fractures was performed based on their locations and displacements. The classification first divides the pelvis into 4 parts: left half pelvis and right half pelvis; sacral Denis Ⅲ area and pubic symphysis. The symmetrical left and right sacral Denis Ⅰ and Denis Ⅱ areas are also included in the left/right half pelvis. Subsequently, the left half pelvis and right half pelvis are divided into 4 regions and marked by capitalized English letters: Sacrum Area (including Denis Ⅰ and Denis Ⅱ, denoted as S), Sacroiliac Joint Area (denoted as J), Iliac Area (denoted as I), and Pubic Area (denoted as P); to distinguish right/left, R and L are used as prefixes. The 2 asymmetric parts are also marked with English letters: Denis Ⅲ area of the sacrum (denoted as Sac), and pubic symphysis (denoted as C). Afterwards, the fracture line morphology and displacement in each region are marked digitally to form a complete JST classification system. The inter- and intra-observer reliabilities (Fleiss' and Cohen's Kappa) of the JST classification system were tested by 3 observers with more than 10 years of experience in pelvic fracture treatment.Results:Consistency analysis of the JST classification results showed that the mean κ value of the intra-observer reliability was 0.818 (from 0.658 to 0.946, P<0.001) and the inter-observer reliability 0.873 (from 0.674 to 1.000, P<0.001), both indicating excellent agreement. Of the 63 patients, 59 obtained successful closed reduction with the assistance of the Rossum Robot R-Universal intelligent orthopedic surgical robot system after fracture classification by the JST system, yielding a success rate of 93.7% (59/63). Conclusions:The new JST classification system for pelvic fractures demonstrates strong intra and inter-observer reliabilities compared with traditional classification systems. As JST classification system labels each fracture site and key bones, it is of great significance for the deep learning and intraoperative operations of intelligent fracture robots.
7.Inhibition effect of temozolomide on proliferation of human glioma cells by autophagy-induced pyroptosis
Shiwen LYU ; Zhun HUANG ; Yalan ZHU ; Chunxia HONG
Chinese Journal of Endocrine Surgery 2023;17(5):624-629
Objective:To investigate the effect of temozolomide on autophagy of human glioma cells, and the inhibition of autophagy induced pyrocytosis on the proliferation of human glioma cells.Methods:2-64 μ mol/L of temozolomide was used to treat glioma U251 cells cultured in vitro. MTT assay was used to detect cell viability, MDC staining was used to detect autophagic vesicles in cells, cloning assay was used to detect cell proliferation, RT qPCR was used to detect the expression level of pyroptosis related mRNA in cells, Western blot was used to detect the expression of autophagy related proteins and pyroptosis related proteins in cells, and the relationship between autophagy and pyroptosis was detected by adding autophagy inhibitors.Results:Temozolamide could induce autophagy of human glioma cells, and significantly induce tumor cells to pyroptosis, thereby inhibiting the proliferation of tumor cells; RT qPCR results showed that caspase-1, GSDMD, IL-1 after temozolomide administration compared with the normal group β, the mRNA expression levels of IL-18 and NLRP3 increased significantly; Western blot results showed that Cleaved-caspase-1, Cleaved-N-terminalGSDMD, IL-1 β、IL-18 and NLRP3 protein were up-regulated; The incidence of pyroptosis decreased after the addition of autophagy inhibitors.Conclusion:Temozolamide can induce autophagy of human glioma cells, and then lead to pyroptosis, which plays an inhibitory role in proliferation.
8.The effect of the quality improvement project of admission hypothermia on short-term outcomes of preterm infants less than 34 weeks gestational age
Fengna ZHU ; Ziqi WU ; Sicong PENG ; Ying XIA ; Shiwen XIA
Chinese Pediatric Emergency Medicine 2023;30(3):188-193
Objective:To study the high risk factors of hypothermia in premature infants with gestational age ≤34 weeks, and to analyze the incidence of hypothermia before and after the implementation of the quality improvement program of hypothermia in hospital and its influence on various systemic complications, aiming to improve the early identification of hypothermia and to reveal the important clinical significance of temperature management in time.Methods:Clinical data of preterm infants born in Maternal and Child Health Hospital of Hubei Province from May 2017 to December 2018, with gestational age ≤34 weeks, and admitted within 1 hour after birth were collected.According to the admission temperature, the infants were divided into normal temperature group (36.5-37.5 ℃), mild hypothermia group (36.0-36.4 ℃), moderate hypothermia gsroup (32.0-35.9 ℃), and severe hypothermia group (<32.0 ℃). The high risk factors of hypothermia in premature infants were analyzed.The incidence and degree of hypothermia and the effects on the systemic complications before and after the implementation of the hypothermia quality improvement program were compared.Results:A total of 306 premature infants were enrolled in the study, including 63(20.6%)cases in the normal temperature group, 115(37.6%) cases in the mild hypothermia group, and 128(41.8%) cases in the moderate hypothermia group, without severe hypothermia.Infants with birth asphyxia were at higher risk for hypothermia( OR=0.195, 95% CI 0.046-0.833, P=0.027); the lower the Apgar score at 1 min( r=0.123, P=0.032)and 5 min after birth( r=0.136, P=0.017), the higher the risk of admission hypothermia.After the quality improvement project, the incidence of admission hypothermia decreased from 82.3% to 73.8%( χ2=32.67, P<0.001), and the use of pulmonary surfactant in infants with respiratory distress syndrome was significantly reduced(70.0% vs. 32.0%, χ2=40.11, P<0.001), and the incidence of hypotension within 72 hours after birth decreased(11.8% vs. 4.9%, χ2=3.87, P<0.049). Conclusion:Birth asphyxia is a risk factor for admission hypothermia in premature infants, and Apgar score is associated with admission hypothermia in premature infants.Temperature management of preterm infants can significantly reduce the incidence of hypothermia and hypotension, and reduce the use of pulmonary surfactant in respiratory distress syndrome infants.
9.Progress in minimally invasive techniques for pelvic fractures
Chinese Journal of Orthopaedic Trauma 2023;25(6):518-522
Minimally invasive treatment is the development trend in surgical techniques for pelvic fractures. There have been numerous minimally invasive techniques for pelvic fractures, each having its own advantages. This paper reviews the literature recently published at home and abroad concerning minimally invasive treatment of pelvic fractures so as to summarize the existing minimally invasive techniques for the fractures and their supporting theories, hoping traumatic orthopedists to know more about the minimally invasive techniques. As a result, Chinese traumatic orthopedists may formulate personalized treatment protocols to suit the specific condition of each patient so that the surgical invasion can be minimized and the intraoperative and postoperative complications reduced.
10.Effects of weekend admission on total hospitalization expenses for elderly patients with hip fracture under the geriatric orthopedic co-management
Lifang WANG ; Yanwei LYU ; Yufeng GE ; Lei PAN ; Fangfang DUAN ; Shiwen ZHU
Chinese Journal of Orthopaedic Trauma 2023;25(12):1049-1055
Objective:To explore the impact of weekend hospitalization on total hospitalization expenses for elderly patients with hip fracture under the geriatric orthopedic co-management.Methods:A retrospective analysis was conducted to analyze the clinical data of elderly patients with hip fracture who had been hospitalized for surgical treatment at Beijing Jishuitan Hospital from May 2015 to December 2020. They were divided into 2 groups based on their admission date. Group A was admitted from Monday to Thursday while Group B from Friday to Sunday. The general demographic data, diagnostic information, comorbidities, hospitalization expenses of the patients were collected. The differences in total hospitalization expenses, hospitalization time, rate of surgery within 48 hours and rate of hospital mortality between the 2 groups were analyzed by rank sum test, chi square test, correlation analysis, and multiple linear regression.Results:A total of 6,075 patients with hip fracture were included in this study, including 1,675 males and 4,400 females with a median age of 80 (74, 85) years. There were 3,935 ones in group A and 2,140 ones in group B. The total hospitalization expenses for group A was 58,160.52 (49,215.45, 72,748.94) yuan, insignificantly lower than those for Group B [58,412.90 (49,163.58, 72,712.61) yuan] ( P>0.05). The rate of surgery within 48 hours for group A was 75.8% (2,984/3,935), significantly higher than that for group B [49.3% (1,054/2,140)]. The hospitalization time for group A was 5 (4, 7) days, significantly less than that for group B [5 (4, 7) days] ( P<0.05). There was no significant difference in the rate of hospital mortality between the 2 groups ( P>0.05). Multiple linear regression analysis showed that total hospitalization expenses were significantly higher for patients admitted on weekends, hospitalization time was positively correlated with total hospitalization expenses, and total hospitalization expenses were significantly lower for the patients undergoing surgery within 48 hours ( P<0.05). Conclusion:Admission on weekends can increase total hospitalization expenses, prolong hospitalization time, and reduce rate of surgery within 48 hours for elderly patients with hip fracture.

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