1.Transcranial direct current stimulation at different targets for Parkinson's disease:a network Meta-analysis
Yulin YANG ; Wanpeng CHANG ; Jiangtao DING ; Hongli XU ; Xiao WU ; Boheng XIAO ; Lihong MA
Chinese Journal of Tissue Engineering Research 2024;28(11):1797-1804
OBJECTIVE:To systematically evaluate the efficacy of transcranial direct current stimulation on the motor function of patients with Parkinson's disease,and to compare the efficacy of transcranial direct current stimulation at different targets on the motor function of patients with Parkinson's disease,so as to provide a theoretical basis for the target selection of transcranial direct current stimulation in clinical practice. METHODS:Cochrane Library,PubMed,Web of Science,CNKI,VIP,WanFang Data were retrieved for randomized controlled trials on the improvement of motor function in patients with Parkinson's disease by transcranial direct current stimulation published from the database inception to January 2023.The keywords were"Parkinson,transcranial direct current stimulation"in English and Chinese.The quality of the included studies was evaluated using the Cochrane 5.1.0 risk of bias assessment tool and the PEDro scale.Meta-analysis of outcome indicators was performed using RevMan 5.4 and Stata 17.0 software. RESULTS:Fifteen randomized controlled trials were finally included,and the PEDro scale showed that all were high-quality or very high-quality studies.Meta-analysis showed that transcranial direct current stimulation significantly improved Unified-Parkinson Disease Rating Scale part III score[mean difference(MD)=-2.49,95%confidence interval(CI):-4.42 to-0.55,P<0.05),step frequency score(MD=0.07,95%CI:0.03-0.11,P<0.05)and step speed score(MD=0.02,95%CI:0.00-0.05,P<0.05),but not for Berg Balance Scale scores(MD=2.57,95%CI:-0.74 to 5.87,P>0.05).Network Meta-analysis probability ranking:In terms of Unified-Parkinson Disease Rating Scale part III scores,the probability ranking results of target stimulation efficacy were dorsal lateral prefrontal cortex(52.4%)>primary motor cortex(45.8%)>central point of the brain(1.8%)>conventional rehabilitation(0%);in terms of gait frequency scores,the probability probability ranking results of target stimulation efficacy were cerebellum(50.1%)>central point of the brain(45.8%)>dorsal lateral prefrontal cortex(3.9%)>primary motor cortex(0.2%)>conventional rehabilitation(0%);in terms of gait speed scores,the probability ranking results of target stimulation efficacy were cerebellum(64.8%)>dorsal lateral prefrontal cortex(23.8%)>central point of the brain(9.4%)>primary motor cortex(1.7%)>conventional rehabilitation(0.4%);in terms of Berg Balance Scale scores,the probability ranking results of target stimulation efficacy were cerebellum(77.4%)>dorsal lateral prefrontal cortex(20.7%)>central point of the brain(0.7%)>conventional rehabilitation(0.2%). CONCLUSION:Transcranial direct current stimulation significantly improves motor function of patients with Parkinson's disease,with better motor coordination in the dorsolateral prefrontal cortex and better walking and balance in the cerebellum.
2.Content Determination of Alkaloids and Catalpol in Different Proportions of Qianjin Huanglian Pills
Hailong MAO ; Rui BAO ; Xiao WAN ; Hongli YAN
Herald of Medicine 2024;43(4):601-606
Objective To establish an HPLC method for the determination of alkaloids(epiberberine,coptisine,palma-tine,berberine)and catalpol in different ratios(1∶1,1∶10)of ancient and modern Qianjin Huanglian Pills,and to compare the differences in their contents.The content differences were compared to preliminarily evaluate the differences in the efficacy of Qianjin Huanglian Pills in the treatment of diabetes under different preparation processes and different ratios.Methods The alkaloid solvent was methanol∶ hydrochloric acid(100∶1).The detection conditions were as follows:C18 column,acetonitrile-0.05 mol·L-1 potassium dihydrogen phosphate solution(50∶50),detection wavelength 345 nm,column temperature 30℃,flow rate 1 mL·min-1,injection volume 10 μL.The catalpol solution was methanol∶ water(20∶80).The detection conditions were as follows:chromatographic column C18 column,methanol-0.1%phosphoric acid solution(1∶ 99),detection wavelength 210 nm,column temperature 30℃,flow rate 1 mL·min-1,injection volume 10 μL.Results The established method was spe-cific,and the separation effect of the five components was good.It exhibited a good linear relationship(R2>0.999)in their respec-tive linear ranges.The repeatability,precision,stability,and sample recovery rate all met the requirements.The content of four alka-loids in the ancient method 1∶1 was the highest,and the content of catalpol was the lowest.The content of four alkaloids in the ancient method 1∶10 was the lowest;the content of 1∶1 in the present method was higher than that in the ancient method 1∶10,and the content of berberine in the present method 1∶10 was slightly lower than that in the present method 1∶1,and the rest were higher than that in the present method 1∶1.The PCA results showed that the chemical composition contents of the four kinds of Qianjin Huanglian pills were very different.Conclusion The method is simple,accurate,and reproducible,making it suitable for the quality control of Qianjin Huanglian Pills.It provides a theoretical basis for exploring the difference in efficacy of Qianjin Huanglian Pills.
3.miR-362-3p inhibited the invasion and metastasis of oral squamous cell carcinoma cells by targeting the regula-tion of pituitary tumor-transforming gene 1
Xiao DING ; Jiawen CHEN ; Pengyu QU ; Chenyu SUN ; Hongli LI ; Wenting HU ; Xin FAN
West China Journal of Stomatology 2024;42(1):46-55
Objective This study aimed to explore the effect of pituitary tumor-transforming gene 1(PTT-G1)on the invasion and proliferation of oral squamous cell carcinoma(OSCC)cell lines under the action of miR-362-3p.Methods The bioinformatics online database was used to query the expression of PTTG1 in head and neck squamous cell carcinoma(HNSCC).The expression of PTTG1 in the Cal-27,HN-30,and HOK cell lines was detected by Western blot.A wound-healing assay was used to determine the effect of PTTG1 on the migration ability of the OSCC cells.The Transwell assay was used to examine the changes in cell-invasion ability.5-ethynyl-2'-deoxyuridine(EdU)cell-proliferation assay was used to detect changes in cell-proliferation ability.Bioinformatics approach predicted the upstream miRNA of PTTG1.The targeting relationship between miR-362-3p and PTTG1 was examined by the dual luciferase assay,and quantitative real-time polymerase chain reaction(qRT-PCR)was used to determine the expression of miRNA in OSCC tissues.Results The ENCORI database showed that PTTG1 expression was up-regulated in OSCC tissues.Western blot confirmed that PTTG1 expression was up-regulated in Cal-27 and HN-30 cells than HOK cells.PTTG1 knockout can inhibit the migration,invasion,and prolif-eration of Cal-27 and HN-30 cells(P<0.05).Bioinformatics prediction websites predicted that the upstream miRNA of PTTG1 was miR-362-3p,and PTTG1 can bind to miR-362-3p.Results of qRT-PCR showed that miR-362-3p expression was downregulated in OSCC tissues compared with normal tissue(P<0.05).Transwell and EdU experiments confirmed that miR-362-3p knockdown can promote the invasion and proliferation of Cal-27 and HN-30 after PTTG1 knockdown.Conclusion miR-362-3p can inhibit the invasion and proliferation of Cal-27 and HN-30 cells by targeting PTTG1.
4.Mediating role of self-regulatory fatigue between psychological distress and patient activation among old patients receiving PCI
Wenjing ZHANG ; Feifei GUO ; Zhihao WANG ; Hongli GUO ; Yunling XIAO
Chinese Journal of Geriatrics 2024;43(8):1025-1029
Objective:To investigate the mediating role of self-regulatory fatigue between psychological distress and patient activation among elderly patients undergoing PCI, in order to establish a foundation for enhancing patient activation in this population.Methods:A cross-sectional study was conducted on 267 elderly patients hospitalized in the Department of Geriatrics at Qilu Hospital, Shandong University, for PCI treatment of coronary artery disease from March to August 2023.The study utilized the Patient Self-Regulation Fatigue Scale(SRF-S), the Positivity Scale(PAM), and the Kessler Scale of Psychological Distress(K10)to evaluate self-regulation fatigue, positivity, and psychological distress in the patients.Additionally, a structural equation model was developed to investigate the relationships among these variables.Results:The study found that the scores of PAM, SRF-S, and K10 were as follows: (56.49±9.52), (42.74±8.11), and(23.42±4.45)respectively.Patient activation showed a negative correlation with self-regulatory fatigue and psychological distress, while self-regulatory fatigue was positively correlated with psychological distress.Furthermore, self-regulatory fatigue was identified as playing a partial mediating role between psychological distress and patient activation, with a mediating effect of 0.198, which accounted for 53.8% of the total effect.Conclusions:self-regulatory fatigue serving as a mediating factor between psychological distress and patient activation.Clinicians should prioritize addressing self-regulatory fatigue to enhance the efficacy of patient activation interventions.
5.Effect of miR-1298-5p on biological behavior of non-small cell lung cancer cells and tumor immune microenvironment by targeting MSH2 gene
Yaosheng ZHANG ; Xiuli YANG ; Xiao REN ; Hongli WANG ; Ling SHEN ; Guosheng HUANG
Chinese Journal of Immunology 2024;40(9):1889-1894,1901
Objective:To investigate the potential mechanism of miR-1298-5p in non-small cell lung cancer(NSCLC)to regu-late the MSH2 gene and its effect on the biological behavior of tumor cells and the tumor immune microenvironment.Methods:Bioin-formatics was used to identify the key genes and miRNA involved in NSCLC.Cell proliferation was detected by CCK-8 assay,and cell invasion and migration were detected by Transwell assay.Levels of inflammatory factors were detected by ELISA assay.Western blot was used to measure the expression of MSH2 in cells,and fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expressions of miR-1298-5p and MSH2 gene in NSCLC cells.Dual luciferase reporter gene assay was performed to verify the targeting relationship between miR-1298-5p and MSH2.Spearman correlation analysis was performed to correlate miR-1298-5p with immune cells and immune factors in the tumor immune microenvironment.Results:The level of miR-1298-5p was down-regulated in NSCLC cells compared with normal lung tissue cells.miR-1298-5p overexpression inhibited the proliferation,migration and inva-sion of NSCLC cells.MSH2 was confirmed to be a target gene of miR-1298-5p using luciferase reporter gene assay.Furthermore,down-regulation of miR-1298-5p in NSCLC cells could be reversed by silencing MSH2.miR-1298-5p expression levels were negatively corre-lated with the levels of Treg,IL-10,and TGF-β,and positively correlated with the levels of CD3+T,CD4+T,CD8+T,NK cells,IL-2,and IFN-γ.Conclusion:miR-1298-5p negatively regulates MSH2 to inhibit the proliferation,invasion and migration of NSCLC cells and improve the tumor immune microenvironment.
6.Expression of severe fever with thrombocytopenia syndrome virus Gn-D Ⅲ-Ⅲ and development of indirect ELISA for antibody detection
Mengyao ZHANG ; Tianlai LIANG ; Feihu YAN ; Tao CHEN ; Cuicui JIAO ; Hongli JIN ; Jiaoyan LUAN ; Xiao WU ; Pei HUANG ; Haili ZHANG ; Qin NING ; Hualei WANG ; Yuanyuan LI
Chinese Journal of Veterinary Science 2024;44(8):1704-1712
The PCR-amplified severe fever with thrombocytopenia syndrome virus(SFTSV)Gn-DⅢ-Ⅲ gene was inserted into the pET-30a(+)prokaryotic expression vector to generate the re-combinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ.The plasmid was transformed into E.coli BL21(DE3)for Gn-DⅢ-m protein expression and the expression conditions were optimized.The Gn-DⅢ-Ⅲ protein purified with Ni-NTA column affinity chromatography was applied as the captured antigen to establish an indirect ELISA method for the detection of SFTSV antibody.The results demonstrated that the recombinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ was successfully constructed as identified by PCR and sequencing.The recombinant protein SFTSV Gn-D m-Ⅲ was soluble ex-pression in E.coli under the optimal induction conditions of 0.4 mmol/L IPTG at 25 ℃ for 4 h,and the protein purity was 91.77%after purification by Ni-NTA column.The optimal reaction con-ditions for the indirect ELISA of SFTSV antibody were as follows:coating antigen concentration(5 μg/mL),primary antibody(incubation at 37 ℃ for 1.5 h),and secondary antibody(diluted 1:10 000 and incubated at 37 ℃ for 1 h).The established method had no cross-reactivity with Rift Valley fever virus(RVFV),Ebola virus(EBOV),and tick-borne encephalitis virus(TBEV)posi-tive sera.The method had a high sensitivity,with P/N>2.1 for SFTSV-positive sera diluted to 81920.Coefficients of variation for intra-and inter-batch reactions were less than 10%.Detection of four SFTSV-infected human clinical serum samples showed the serum samples from patients in re-mission were tested as positive(P/N>2.1),while serum samples from patients with multiple or-gan failure were detected as negative(P/N<2.1).The results indicated that the SFTSV Gn-D Ⅲ-Ⅲ protein was successfully expressed and purified,and it was used as the coating protein to estab-lish an indirect ELISA assay for SFTSV antibody,which possesses good specificity,sensitivity and reproducibility.This method might be applied to detect human SFTSV clinical serum samples.
7.Exosomes Derived from Mouse Breast Carcinoma Cells Facilitate Diabetic Wound Healing
Chao ZHANG ; Wenchi XIAO ; Hao WANG ; Linxiao LI ; Yan YANG ; Yongwei HAO ; Zhihao XU ; Hongli CHEN ; Wenbin NAN
Tissue Engineering and Regenerative Medicine 2024;21(4):571-586
BACKGROUND:
Exosomes derived from breast cancer have been reported to play a role in promoting cell proliferation, migration, and angiogenesis, which has the potential to accelerate the healing process of diabetic wounds. The aim of this investigation was to examine the function of exosomes originating from 4T1 mouse breast carcinoma cells (TEXs) in the process of diabetic wound healing.
METHODS:
The assessment of primary mouse skin fibroblasts cell proliferation and migration was conducted through the utilization of CCK-8 and wound healing assays, while the tube formation of HUVECs was evaluated by tube formation assay. High-throughput sequencing, RT-qPCR and cell experiments were used to detect the roles of miR-126a-3p in HUVECs functions in vitro. The in vivo study employed a model of full-thickness excisional wounds in diabetic subjects to explore the potential therapeutic benefits of TEXs. Immunohistochemical and immunofluorescent techniques were utilized to evaluate histological changes in skin tissues.
RESULTS:
The findings suggested that TEXs facilitate diabetic wound healing through the activation of cell migration, proliferation, and angiogenesis. An upregulation of miR-126a-3p has been observed in TEXs, and it has demonstrated efficient transferability from 4T1 cells to HUVEC cells. The activation of the PI3K/Akt pathway has been attributed to miR-126a-3p derived from TEXs.
CONCLUSIONS
The promotion of chronic wound healing can be facilitated by TEXs through the activation of cellular migration, proliferation, and angiogenesis. The activation of the PI3K/Akt pathway by miR-126a-3p originating from TEXs has been discovered, indicating a potential avenue for enhancing the regenerative capabilities of wounds treated with TEXs.
8.Correlation between paraspinal muscle atrophy, morphological changes of facet joints and adjacent segment degeneration after lumbar fusion
Dachuan LI ; Xiao LU ; Guangyu XU ; Jian SONG ; Minghao SHAO ; Feizhou LYU ; Xiaosheng MA ; Xinlei XIA ; Hongli WANG ; Jianyuan JIANG
Chinese Journal of Orthopaedics 2022;42(19):1292-1300
Objective:To investigate the correlation between paraspinal muscle atrophy, morphological changes of facet joints and adjacent segment disease (ASDis) after lumbar fusion operation.Methods:A retrospective study was conducted among 195 patients who underwent posterior lumbar fusion again for ASDis at this institution from January 2014 to December 2020, including 29 patients with ASDis whose initial surgical fusion segment was L 4,5. According to Roussouly's staging, there were 5 cases of type I, 9 cases of type II, 10 cases of type III, and 5 cases of type IV. Another 29 cases were selected from patients without ASDis after lumbar fusion as a control group. The control group was paired 1∶1 with the ASDis group according to gender, fusion segment, and Roussouly typing of the lumbar spine. The cross-sectional area (CSA) and fat infiltration (FI) of paravertebral muscle, facet joint angle (F-J) and pedicle facet (P-F) angle before the first (second) operation were measured and compared between the two groups. Then logistic regression analysis was used to determine the predictors of ASDis after posterior lumbar fusion. Finally, the receiver operation characteristic (ROC) curve was described, and the area under the curve (AUC) and cut-off point were calculated. At the same time, the paraspinal muscle atrophy before the second operation in ASDis group was measured. Results:The average follow-up time of 98 patients was 59.25±6.38 months (range, 49-73 months). The average body mass index (BMI) of ASDis group was 24.76±3.64 kg/m 2, which was higher than that in control group (22.24±2.92 kg/m 2) ( t=2.481, P=0.041). The average CSA and relative cross-sectional area (rCSA) of paraspinal muscle in ASDis group were 3 214.32± 421.15 mm 2 and 1.69±0.36 respectively, which were less than 3 978.91±459.87 mm 2 and 2.26±0.29 in control group ( t=10.22, P=0.012; t=9.47, P=0.038). The FI degree of paraspinal muscle in ASDis group (21.95%±5.89%) was significantly higher than that in control group (14.64%±7.11%) ( t=7.32, P=0.002). The F-J angle in ASDis group was 35.06°±3.45°, which was less than 38.39°±4.67° in control group ( t=4.76, P=0.027). The P-F angle in ASDis group was 117.39°±8.13°, which was greater than 111.32°±4.78° in control group ( t=5.25, P=0.031). Multivariate logistic regression analysis showed that higher BMI ( OR=1.34, P=0.038), smaller rCSA of paraspinal muscle ( OR=0.02, P=0.017) and higher FI of paraspinal muscle ( OR=1.58, P=0.032) were the risk factors of postoperative ASDis. The ROC curve showed that the AUC of BMI was 0.680 and the cut-off point was 22.58 kg/m 2; The AUC of the FI of paraspinal muscle was 0.716 and the cut-off point was 15.69%; The AUC of rCSA of paraspinal muscle was 0.227 and the cut-off point was 1.92. For ASDis patients, the paraspinal muscle before the second operation had a higher degree of FI (25.47%±6.59% vs. 21.95%±5.89%, t=3.99, P=0.042) and a smaller rCSA (1.52±0.28 vs. 1.69±0.36, t=3.85, P=0.038) than that before the first operation. The difference between the FI degree of paraspinal muscle before the second operation and the first operation was negatively correlated with the occurrence time of ASDis ( r=-0.53, P=0.039) , and the difference of rCSA was positively correlated with the occurrence time of ASDis ( r=0.64, P=0.043) . Conclusion:When BMI >22.58 kg/m 2, FI of paraspinal muscle >15.69%, and rCSA of paraspinal muscle <1.92, it suggests that ASDis is more likely to occur after operation. And the more obvious paraspinal muscle atrophy after the first operation, the earlier ASDis may occur. Morphological changes of facet joints cannot be used as an index to predict the occurrence of ASDis.
9.Effect of cage height on adjacent segment degeneration during oblique lumbar interbody fusion: a 3D finite element study
Xiao LU ; Fei ZOU ; Feizhou LYU ; Xiaosheng MA ; Xinlei XIA ; Hongli WANG ; Jianyuan JIANG
Chinese Journal of Orthopaedics 2022;42(19):1301-1311
Objective:To investigate the influence of interbody cage height during oblique lumbar interbody fusion (OLIF) on lumbar biomechanics with different degrees of degeneration and to provide a reference for cage choice.Methods:The finite element model of normal lower lumbar spine (L 3-S 1) was built and validated, then constructed three different degenerative segments in L 3, 4, and the cages with different height (8, 10,12, 14 mm) were implanted into L 4, 5 disc. All the twelve models were loaded with pure moment of 7.5 N·m to produce flexion, extension, lateral bending and axial rotation motions on lumbar spine, and the effects of cage height on range of motion (ROM), intervertebral pressure in adjacent segments and stress in facet joints were investigated. Results:The ROM of adjacent segments and the maximum stress of intervertebral discs increased with the increase of cage height, but this trend was not obvious in moderate and severe degeneration groups. After implantation of 4 different height cages (8, 10, 12, 14 mm), the ROM of L 3, 4 segment reached the maximum during extension. The ROM of mild degeneration group was 2.68 °, 2.71 °, 2.94 °, 2.98 °, moderate degeneration group was 2.33°, 2.37°, 2.41°, 2.49°, and severe degeneration group was 1.94 °, 1.99 °, 2.14 °, 2.21 °. The stress of L 3, 4 intervertebral disc reached the maximum during right bending. The maximum stress of L 3, 4 intervertebral disc was 23.95 MPa, 24.60 MPa, 24.90 MPa and 25.34 MPa in mild group, 25.57 MPa, 25.60 MPa, 25.82 MPa and 25.89 MPa in moderate group, and 25.95 MPa, 25.99 MPa, 26.48 MPa and 27.13 MPa in severe group. The maximum stress of L 3, 4 facet joint was 15.87 MPa, 15.78 MPa, 16.29 MPa and 16.43 MPa in mild group, 15.97 MPa, 16.31 MPa, 16.53 MPa and 16.79 MPa in moderate group, and 16.17 MPa, 16.49 MPa, 16.95 MPa and 17.35 MPa in severe group. Conclusion:For patients with mild lumbar degeneration requiring OLIF surgery, the intervertebral height of the surgical segment should not be overstretched. But for patients with moderate to severe lumbar degenerative disease who need to undergo OLIF surgery, it is recommended that the cage height be 0-2 mm higher than the original intervertebral space height.
10.Effect of vagus nerve stimulation by electroacupuncture on GFAP and NeuN in hippocampal of juvenile rats with depression induced by chronic unpredictable stress
Qiuxiang NING ; Hongli PENG ; Jianying XIAO ; Chao YANG ; Zhiqiang ZHOU ; Fang LIU ; Xiaoli LIANG ; Dong YANG
Journal of Chinese Physician 2022;24(4):538-542
Objective:To analyze the expression of glial fibrillary acidic protein (GFAP) and neuronal nuclei (NeuN) antigen in hippocampus based on the depression model of juvenile rats caused by chronic unpredictable stress (CUS), and to explore the effect of electroacupuncture vagus nerve on CUS depression model.Methods:Six juvenile SD rats were selected as the control group (without any stimulation), and the rest were divided into CUS group, pseudo stimulation group, fluoxetine group and electroacupuncture group by random number method after CUS modeling, with 6 rats in each group. Fluoxetine group was given 10 mg/kg fluoxetine intervention; control group and CUS group were given the same amount of normal saline intervention; In the electroacupuncture group, the distal vagus nerve was stimulated after ligation, while in the pseudo stimulation group, only vagus nerve was isolated without electrical stimulation. After 28 d of intervention, the five groups were subjected to Open-field Test and Sucrose Preference Test. Hippocampal neurons were detected by hematoxylin and eosin (HE) staining, and the expressions of GFAP and NeuN in hippocampal were detected by immunohistochemistry.Results:After CUS modeling and before intervention, the number of vertical and horizontal movements, sucrose consumption and sucrose preference in CUS group, pseudo stimulation group, fluoxetine group and electroacupuncture group were significantly lower than those in the control group (all P<0.01); After the intervention, the above indexes in CUS group and pseudo stimulation group were still lower than those in the control group (all P<0.01), but the above indexes in fluoxetine group and electroacupuncture group were significantly higher than those in CUS group and pseudo stimulation group (all P<0.01). HE staining showed that the arrangement of hippocampal neurons in CUS group and pseudo stimulation group were loose, and there were cell swelling and pyknosis, which was significantly improved in fluoxetine group and electroacupuncture group. Immunohistochemical results showed that compared with the control group, the expression of GFAP increased and NeuN decreased in the hippocampus of CUS group and pseudo stimulation group (all P<0.01); Compared with CUS group and pseudo stimulation group, the expression of GFAP decreased and NeuN increased in fluoxetine group and electroacupuncture group (all P<0.01). Conclusions:Electroacupuncture of vagus nerve can obviously improve the depression symptoms of juvenile rats, which is similar to fluoxetine, and may be related to regulating the expression of GFAP and Neun in hippocampus.

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