1.Mechanism of Gushining Granules in Attenuating Dexamethasone-induced Apoptosis of Bone Marrow Mesenchymal Stem Cells via Activating PI3K/Akt/Bad Signalling Pathway
Chengyu CHU ; Lei ZHU ; Long LIANG ; Feng WANG ; Xuejian YU ; Wenwu LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):115-122
ObjectiveTo establish steroid-induced osteonecrosis of the femoral head (SANFH) cell model by using dexamethasone (DEX)-induced bone marrow mesenchymal stem cells (BMSCs) and demonstrate that Gushing Granules (GSNs) exert an improving effect by activating the phosphatidylinositol-3-kinase/protein kinase B/B-lymphoma-2 gene related promoter (PI3K/Akt/Bad) signalling pathway. MethodsFirstly, SD rats were orally administered with drugs at a dose of 0.9 g·kg-1 to prepare GSN-containing serum, and CCK-8 screening was used to determine the optimal dosage and duration of action. Then, BMSCs were cultured and treated with 1×10-6 mol·L-1 DEX, 10% GSN-containing serum, and inhibitor LY294002 of PI3K/Akt signalling pathway for 24 hours to model and group SANFH cells. Cell viability and proliferation were detected by using CCK-8 assay kit and EdU staining kit. Flow cytometry was used to detect cell apoptosis. An alkaline phosphatase (ALP) assay kit was employed to detect ALP expression. In order to detect the PI3K/Akt/Bad signalling pathway and protein and mRNA expression of apoptosis-related proteins such as apoptosis regulatory factors B-cell lymphoma-2 gene (Bcl-2), and Bcl-2-associated X protein (Bax), osteocalcin (OCN), and Collagen Ⅰ, we used Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultsThe CCK-8 assay kit determined that the optimal dosage for GSN-containing serum is 10%, and the duration of action is 48 hours. After modelling and grouping the cells in each group, the detection results showed that the SANFH model group had significantly lower cell viability, cell proliferation, and ALP expression, as well as protein and mRNA expressions of PI3K, Akt, Bad, Bcl-2, OCN, and Collagen I compared to the blank group. The nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry significantly increased (P<0.05,P<0.01). After treatment with GSN-containing serum, cell viability, cell proliferation, and ALP expression, as well as expressions of PI3K, Akt, Bad, Bcl-2, OCN, and Collagen Ⅰ nucleic acids and proteins were significantly increased, while the nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry significantly decreased(P<0.05,P<0.01). Compared with the GSN drug-containing serum group, the simultaneous treatment with the inhibitor LY294002 and GSN drug-containing serum reversed the improvement effect of GSN. Specifically, the cell viability, cell proliferation, ALP expression, and the nucleic acid and protein levels of PI3K, Akt, Bad, Bcl-2, OCN, and Collagen Ⅰ were all significantly decreased, while the nucleic acid and protein levels of the Bax index and the cell apoptosis rate detected by flow cytometry were significantly increased (P<0.05, P<0.01). ConclusionGSNs antagonize DEX-induced apoptosis of BMSCs by activating the PI3K/Akt/Bad signalling pathway, providing a scientific theoretical basis for the clinical treatment of SANFH with GSNs.
2.A clinical study on the treatment of refractory tennis elbow with arc-edge needle therapy based on zaodian theory
Xuejian MA ; Dongzhe ZHANG ; Chaoyang NIU ; Shuaigang DU ; Wuyang WANG ; Shilong CHENG ; Yingyuan XIAO ; Xuechang WANG
The Journal of Practical Medicine 2024;40(15):2161-2165
Objective To explore the clinical efficacy of arc-edge needle therapy based on Zaodian theory in the treatment of refractory tennis elbow.Methods 64 patients with refractory tennis elbow were collected and randomly divided into a control group and an observation group using a random number table method.Among them,32 cases in the control group were treated with local pain point injection.32 patients in the observation group were treated with arc-edge needle therapy once a week,with two treatments per group.The Number rating scale(NRS)Mayo,elbow joint performance score(MEPS)and Disabilities of the arm,shoulder and hand(DASH)score were recorded and compared between the two groups before treatment and at 1,2,4 and 24 weeks of follow-up after treatment,disease efficacy score(WDES)at 24 weeks of follow-up.Results During the treat-ment process,neither group of patients experienced serious adverse reactions.At each stage after treatment,two groups of NRS MEPS,DASH,and WDES at 24 weeks of follow-up all improved compared to before treatment.After one week of treatment,there was no statistically significant difference in NRS scores between the two groups(P>0.05).At follow-up visits at weeks 2,4,and 24,the NRS scores in the observation group were lower than those in the control group.After 1,2,4,and 24 weeks of follow-up after treatment,the DASH score was lower in the observation group than in the control group,and the MEPS score was higher in the observation group than in the control group,with statistically significant differences(P<0.05).After 24 weeks of treatment,the WDES of the observation group was better than that of the control group,and the difference was statistically significant(P<0.05).Conclusion Local pain point injection and arc-edge Zaodian release can alleviate pain and improve elbow joint function in the early stage of refractory tennis elbow.However,the long-term effect of local pain point injection treatment is not good,and the treatment effect of the observation group gradually improves.
3.The application value of MR diffusion tensor imaging in assessing the impact of march training on the thigh muscles of recruits
Yu ZHANG ; Xiaogang WANG ; Wei SONG ; Chao WANG ; Zi MO ; Xuejian ZHANG ; Xiangwei LUO
Journal of Practical Radiology 2024;40(12):2017-2020
Objective To explore the value of MR diffusion tensor imaging(DTI)in evaluating the effect of march training on the thigh muscles of recruits.Methods DTI scans of the right thigh were performed three times in forty recruits:before and after the march training and one month after the rest.Fractional anisotropy(FA)was measured on the cross-sectional images of the thigh muscles,including rectus femoris(RF),vastus medialis(VM),vastus lateralis(VL),vastus intermedius(VI),gracilis muscle(GM),sartorius muscle(SM),semitendinosus muscle(STM),semimembranosus muscle(SMM),long head of biceps femoris(LHBF)and short head of biceps femoris(SHBF).The percentage changes in FA values of each muscle after the training and rest were calculated.Paired samplet-tests were used to analyze the differences in FA among the thigh muscles at different time points,and one-way ANOVA was used to analyze the differences in the percentage changes of FA among the thigh muscles after the training and rest.Results Compared to pre-training,the FA values of all thigh muscles significantly decreased after the training,with statistical differences(P<0.05).After the rest,the FA values of all thigh muscles recovered,but statistical differences remained in RF(P<0.001),VM(P<0.001),VL(P=0.001),STM(P=0.046),and LHBF(P=0.013).After the training and rest,the FA values of the recruits'thigh muscles showed a"decreasing first and then increasing"trend.There were statistical differences in the percentage changes of FA after the training and the recovery percentages of FA after the rest among the thigh muscles(P<0.001,P<0.001).Conclusion DTI may reflect the ultra-structure changes in the thigh muscles of recruits after the march training and provide a quantitative and noninvasive assessment of muscle micro-injuries.
4.Development and evaluation of a mortality risk prediction model for severe bacterial infections in children
Haoyu ZHA ; Rui TAN ; Haonan WANG ; Xuejian MEI ; Mingxing FAN ; Meiling PAN ; Tingting CHEN ; Jun CHEN ; Yao LIU ; Shaodong ZHAO ; Zhuo LI ; Hongjun MIAO
Chinese Journal of Emergency Medicine 2023;32(4):489-496
Objective:To establish a mortality risk prediction model of severe bacterial infection in children and compare it with the pediatric early warning score (PEWS), pediatric critical illness score (PCIS) and pediatric risk of mortality score Ⅲ (PRISM Ⅲ).Methods:A total of 178 critically ill children were selected from the PICU of the Children's Hospital of Nanjing Medical University from May 2017 to June 2022. After obtaining the informed consent of the parents/guardians, basic information such as sex, age, height and weight, as well as indicators such as heart rate, systolic blood pressure and respiratory rate were collected from all children. A standard questionnaire was used to score the child 24 h after admission to the PICU. The children were divided into the survival and death groups according to their survival status at 28 d after admission. A mortality risk prediction model was constructed and nomogram was drawn. The value of the mortality risk prediction model, PEWS, PCIS and PRISM in predicting the risk of death was assessed and compared using the receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC).Results:Among the 178 critically ill children, 11 cases were excluded due to severe data deficiencies and hospitalization not exceeding 24 h. A total of 167 children were included in the analysis, including 134 in the survival group and 33 in the death group. A mortality risk prediction model for children with severe bacterial infection was constructed using pupillary changes, state of consciousness, skin color, mechanical ventilation, total cholesterol and prothrombin time. ROC curve analysis showed that the AUCs of mortality risk prediction model was 0.888 ( P<0.05). The AUCs of PEWS, PCIS and PRISM Ⅲ in predicting death in children with severe bacterial infection were 0.769 ( P< 0.05), 0.575 ( P< 0.05) and 0.759 ( P< 0.05), respectively. Hosmer-Lemeshow goodness-of-fit test showed the best agreement between risk of death and PEWS predicted morbidity and mortality and actual morbidity and mortality (χ 2 = 5.180, P = 0.738; χ 2 = 4.939, P = 0.764), and the PCIS and PRISM Ⅲ predicted mortality rates fitted reasonably well with actual mortality rates (χ 2= 9.110, P= 0333; χ 2 = 8.943, P= 0.347). Conclusions:The mortality risk prediction model for predicting the death risk has better prognostic value than PEWS, PCIS and PRISM Ⅲ for children with severe bacterial infection.
5.Drofenine as a Kv2.1 inhibitor alleviated AD-like pathology in mice through A β/Kv2.1/microglial NLRP3/neuronal tau axis
Jian LU ; Qian ZHOU ; Danyang ZHU ; Xuejian ZHAO ; Yujie HUANG ; Peng CAO ; Jiaying WANG ; Xu SHEN
Chinese Journal of Pharmacology and Toxicology 2023;37(7):546-547
OBJECTIVE Alzheimer disease(AD)is a neurodegenerative disease with clinical hallmarks of pro-gressive cognitive impairment.Synergistic effects of Aβ-tau cascade reaction are tightly implicated in AD patholo-gy,and microglial NLRP3 inflammasome activation drives neuronal tauopathy through microglia and neurons cross-talk.However,the underlying mechanism of how Aβ medi-ates NLRP3 inflammasome remains unclear.Shab related potassium channel member 1(Kv2.1)as a voltage gated po-tassium channel widely distributed in the central nervous system and plays an important role in regulating the out-ward potassium flow in neurons and glial cells.In current work,we aimed to explore the underlying mechanism of Kv2.1 in regulating Aβ/NLRP3 inflammasome/tau axis by using a determined Kv2.1 inhibitor drofenine(Dfe).METHODS Cell-based assays including Western blot-ting and immunofluorescence staining against primary microglia or neurons were carried out to expound the role of Kv2.1 channel in NLRP3 inflammasome activa-tion and subsequent neuronal tau hyperphosphorylation.For animal studies,new object recognition,Y-maze and Morris water maze were performed to evaluate the ame-lioration of Kv2.1 inhibition through either Kv2.1 inhibitor Dfe treatment or adeno-associated virus AAV-ePHP-si-Kv2.1injectionon5×FADADmodel mice.Assays of histol-ogy and immunostaining of tissue sections and Western blotting of brain tissues were performed to verify the con-clusion of cellular assays.RESULTS We reported that oligomeric Aβ(o-Aβ)bound to microglial Kv2.1 and pro-moted Kv2.1-dependent potassium leakage to activate NLRP3 inflammasome through JNK/NF-κB pathway sub-sequently resulting in neuronal tauopathy.Treatment of either Kv2.1 inhibitor Dfe or AAV-ePHP-si-Kv2.1 for brain-specific Kv2.1 knockdown deprived o-A β of its capability in inducing microglial NLRP3 inflammasome activation and neuronal tau hyperphosphorylation,while improved the cognitive impairment of 5×FAD AD model mice.CONCLUSION Our results have highly addressed that Kv2.1 channel is required for o-Aβ driving NLRP3 inflammasome activation and neuronal tauopathy in AD model mice and highlighted that Kv2.1 inhibition is a prom-ising therapeutical strategy for AD and Dfe as a Kv2.1 inhibitor shows potential in the treatment of this disease.
6.Clinical application of adjustable traction skin stretchers in repair of wounds at the lower leg, foot and ankle
Wenchao SONG ; Xuejian WU ; Juyu TANG ; Jianli WANG ; Bo LIU ; Peng XIAO ; Chong MENG ; Jianzhong GUAN ; Tao SONG ; Guohua NI ; Dehong ZHANG ; Weihong ZHANG
Chinese Journal of Orthopaedic Trauma 2023;25(3):226-232
Objective:To evaluate the clinical effects of adjustable traction skin stretchers used in repair of wounds at the lower leg, foot and ankle.Methods:A retrospective study was performed to analyze the clinical data of 56 patients who had been treated for skin defects at the lower leg, foot and ankle from August 2016 to September 2022 at The First Affiliated Hospital of Zhengzhou University, Honghui Hospital, Affiliated to Xi'an Jiaotong University Medical College, The First Affiliated Hospital of Henan Polytechnic University, and Yunnan Zhongde Orthopedic Hospital. There were 35 males and 21 females, aged (39.9±18.7) years. There were 43 traumatic wounds, 3 burns, 6 inflammatory wounds, 3 relief incisions due to osteofascial compartment syndrome, and 1 scar. The areas of skin defect ranged from 2.5 cm × 2.0 cm to 20.0 cm × 10.0 cm. The duration of wounds was (8.6±7.8) d. All the wounds were repaired with adjustable traction skin stretchers. The row-hook type of skin stretchers was used in 28 cases, the single-rod type in 20 cases, the single-rod type combined with an external fixator in 5 cases, and a combination of the row-hook type and the single-rod type in 3 cases.The time for wound traction closure, color of wound skin margin, skin swelling around the wound, functional recovery of affected limb and complications were recorded.Results:The time from skin stretching to wound closure was (7.8±3.8) d in the 56 patients. The color of wound skin edge after stretching was normal in 16 cases, dark red in 38 cases, and dark in 2 cases; the skin swelling around the wound was degree 1 in 21 cases, degree 2 in 33 cases, and degree 3 in 2 cases. The 56 patients were followed up for (8.9±4.1) months. Primary wound closure was achieved in 48 patients, and secondary wound closure in 8 patients after repair with an autologous skin graft. Partial skin necrosis occurred due to tension blisters after skin stretching in 2 patients, one of whom was repaired with an autologous skin graft and the other of whom by dressing change. Deep bone infection recurred in 2 patients whose wounds healed after their bone defects were repaired using Ilizarov technique of bone transfer. In the 56 patients, the muscle strength of the lower extremity beyond the wound was recovered to normal, and the range of motion of the joints adjacent to the wound also recovered to normal.Conclusion:In repair of wounds at the lower leg, foot and ankle, adjustable traction skin stretchers can lead to fine clinical effects and limited complications, because the stretchers can control the tension of skin digitally and precisely.
7.Ilizarov bone transport assisted by a 3D printed patient-specific guide plate for treatment of tibial bone defects
Hao ZHENG ; Lili WANG ; Yong LIU ; Jianli WANG ; Xuejian GAO
Chinese Journal of Orthopaedic Trauma 2023;25(7):617-623
Objective:To evaluate the efficacy of 3D printed patients-specific guide plates in assisting Ilizarov bone transport in the treatment of tibial bone defects.Methods:A retrospective study was conducted to analyze the clinical data of 24 patients with tibial bone defects who had been admitted to Institute of Trauma Orthopedics, The 80th Army Group Hospital of PLA from January 2018 to March 2022. There were 9 males and 15 females with an age of (49.8±6.5) years, and 4 upper tibial defects, 5 middle tibial defects, and 15 lower tibial defects. According to the methods of repairing bone defects, the patients were divided into 2 groups: a 3D printing group of 10 cases where a 3D printed patient-specific guide plate was used to assist Ilizarov bone transport in the treatment of tibial bone defects, and a traditional group of 14 cases where Ilizarov bone transport was performed in a traditional manner. The 2 groups were compared in terms of operation time, frequency of intraoperative fluoroscopy, axial angulation of the tibia at postoperation and the last follow-up, external fixation time (EFT) and external fixation index (EFI). At the last follow-up, healing of bone defects was evaluated according to the criteria of The Association for the Study and Application of the Method of Ilizarov (ASAMI), functional outcomes were evaluated according to the Paley criteria, and needle infection was recorded according to the Paley classification for complications.Results:There was no statistically significant difference in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). All patients were followed up for (11.3±2.0) months on average after operation. The 3D printing group had significantly shorter operation time [(19.9±2.6) min] and significantly lower frequency of intraoperative fluoroscopy [(3.0±0.8) times] than the traditional group [(38.1±2.2) min and (8.9±1.3) times] (P<0.05), and had significantly better axial angulation of the tibia at postoperation and the last follow-up than the traditional group ( P<0.05). There was no significant difference in EFT or EFI between the 2 groups ( P>0.05), and the last follow-up revealed no significant difference either in bone healing, functional outcomes, or needle infection between the 2 groups ( P>0.05). Conclusion:In the treatment of tibial bone defects, compared with conventional Ilizarov bone transport, the Ilizarov bone transport assisted by a 3D printed patient-specific guide plate demonstrates advantages of shorter operation time, lower intraoperative fluoroscopy, and higher reduction accuracy.
8.Analysis of the Symptoms of Jaundice Formula"Ma-Huang Lian-Yao Chi-Xiao-Dou Tang",Examination of Lian-Yao,and Insight into the Medicinal Value and Research of Forsythiae Fructus
Tao WANG ; Qilei CHEN ; Baifang WANG ; Jiankai TANG ; Xuejian GUO ; Shifei LI ; Hubiao CHEN ; Liwei ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(9):2909-2913
Ma-huang Lian-yao Chi-xiao-dou Tang is one of the three formulas for typhoid jaundice.But it has not received much attention,and the source of Lian-yao in the formula is unknown and the status and role are difficult to identify.In this paper,we first analyzed the functions of Ephedra and the symptoms of the whole formula.The formula was mainly used to"clear damp-heat"in order to remove jaundice.On this basis,it is proved that Lian-yao is the root of Forsythia,not"Qiao-gen".Today,the use of Forsythia instead of Lian-yao has the same effect.Then,the indispensable and important position of Forsythia in the formula was analyzed.It also pointed out some valuable directions for the medicinal use and research and development of Forsythia based on its wide range of effects but the lack of main symptoms.This paper provides insights for in-depth theoretical and practical research on Forsythiae Fructus.
9.Thoracoumbilical flap combined with random abdominal flap in repair of large-area soft tissue defects of calf in children
Hao ZHENG ; Yong LIU ; Lili WANG ; Jianli WANG ; Xuejian GAO
Chinese Journal of Microsurgery 2022;45(5):528-533
Objective:To investigate the clinical effect of thoracoumbilical flap combined with random abdominal flap in repair of large-area soft tissue defects of calf in children.Methods:The clinical data of 16 children with large-area soft tissue defects of calf treated with thoracoumbilical flap combined with abdominal random flap from January 2004 to December 2007 in PLA Trauma Orthopaedic Research Institute, 80th Group Military Hospital of the PLA were retrospectively analysed. There were 7 boys and 9 girls aged 8 to 14(11.3 in average) years old. Six cases were crushed by heavy objects, 6 crushed by wheels, 3 by thermal press and 1 by machine strangulation. After thorough debridement, the wound area ranged from 16.0 cm×9.0 cm to 38.0 cm×15.0 cm. Four cases were treated after 3-10 hours of injury by emergency surgery. Twelve cases received surgeries 0-11 days after hospital admission and wound being stabilised. Doppler ultrasound was used to locate the perforating vessels according to the location, size and shape of the wound. Thoracoumbilical flap combined with abdominal random flap were designed and harvested to repair the wound. The sizes of flaps were 18.0 cm×11.0 cm-40.0 cm×16.0 cm. All patients entered follow-up at the outpatient clinic or through WeChat interviews. The appearance, texture of the flap and limb recovery were checked and recorded.Results:After surgery, all of 16 flaps survived, of which 12 flaps had phase-one healing, 3 flaps had small area of necrosis at the edge, which healed after repeated dressing changes and 1 flap developed vascular comproise, and survived after surgical exploration. The donor sites healed in phase-one. All 16 children had 6 months to16 years of follow-up, with an average of 20.7 months. The colour of the flaps was normal with soft texture. The motor function of calf was satisfactory. According to Punor functional evaluation criteria, 12 cases were in excellent and 4 in good.Conclusion:The thoracoumbilical flap combined with abdominal random flap features a reasonable design, strong blood supply and repair of a large area. It is a reliable method for repairing large area soft tissue defects in the calf of children.
10.Characteristic of 12 cases of acute brucellosis of Brucella melitensis in Xinjiang Uygur Autonomous Region
Xuejian ZHANG ; Songsong XIE ; Yuanzhi WANG ; Xiangwei WU
Chinese Journal of Endemiology 2022;41(12):1013-1018
Objective:To identify the biotypes of blood culture positive Brucella in patients with acute brucellosis in Xinjiang Uygur Autonomous Region (hereinafter referred to as Xinjiang), and to learn about the clinical characteristics of these patients. Methods:Among the 30 patients diagnosed with acute brucellosis in First Affiliated Hospital, School of Medicine, Shihezi University from January 2019 to July 2021, the positive strains of blood culture Brucella of 12 patients with acute brucellosis were used to identify the biotype by AMOS-PCR. Then the general and clinical data of the 12 patients were collected, including demographic characteristics, epidemiological characteristics, clinical characteristics, laboratory tests, treatment and prognosis. Results:AMOS-PCR results showed that all 12 strains were Brucella melitensis. Among the 12 patients, 9 were males and 3 were females, all of whom had a clear history of livestock contact, and the onset time was concentrated in April to September. All 12 patients showed fever, followed by headache and dizziness (9/12), weakness (8/12), digestive symptoms (8/12), hyperhidrosis (7/12), lumbago and backache (6/12), and arthralgia (6/12). One patient was complicated with pleural effusion and pericardial effusion. One patient was complicated with pleural effusion, pelvic effusion and ascites. Only ascites occurred in 2 patients. Liver function was abnormal in 3 patients, lactate dehydrogenase increased in 6 patients, and blood creatinine decreased in 9 patients. Among the 12 patients, 10 patients were treated with rifampicin combined with doxycycline. The other 2 patients were treated with levofloxacin and doxycycline, supplemented by symptomatic treatment with hepatoprotective drugs due to impaired liver function. After discharge, the patients received out-of-hospital oral anti-infectious drugs for 1-2 courses (6 weeks each course), and all indicators returned to normal at 6 months and 12 months after discharge. Conclusions:All the 12 patients with acute brucellosis in Xinjiang are infected with Brucella melitensis. In addition to the common clinical features, patients are characterized by decreased creatinine and increased lactate dehydrogenase. The standardized anti- Brucella treatment is still the main treatment strategy.

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