Objective:To explore the effect of transcutaneous electrical nerve stimulation (TENS) on interleukin-33 (IL-33)/growth stimulation expressed gene 2 (ST2) signaling pathway in the dorsal root ganglia (DRGs) of rats modeling hyperalgesia (HP).Methods:This study consisted of two experiments. In the first, 30 Sprague-Dawley (SD) rats were randomly divided into a blank group, a Sham-HP group, an HP group, an antibody group and an inhibitor group, each of 6. HP was induced in all except the rats of the blank and Sham-HP groups by injecting carrageenan (Car) and prostaglandin E2 subcutaneously at the bottom of the left hind feet. The antibody and inhibitor groups were then given intrathecal injections of anti-ST2 antibody and a tumor necrosis factor α (TNF-α)-specific inhibitor, respectively. In the second experiment, 42 SD rats were randomly divided into a Sham-HP group, an HP group, a TENSⅠgroup, a TENS II group, a TENS I inhibitor group, a TENS II inhibitor group, and a Sham-TENS group, each of 6. All of the groups had HP induced as in experiment one. All of the rats except those in the Sham-HP, HP and Sham-TENS groups were then given TENS, and the TENS I and II inhibitor groups were offered intrathecal injection of TNF-α-specific inhibitors. Mechanical pain thresholds (MPTs) were documented 4h, 24h, 48h, 72h, 6d, 7d 4h, 7d 1h, and 7d after the Car injections. Western blotting was used to measure the protein expressions of IL-33, ST2 and TNF-α 6d after the Car injection in both experiments.Results:In experiment one, the average MPTs of the HP, antibody and inhibitor groups had decreased significantly 4 hours after the Car injection compared with the blank and Sham-HP groups. However, 7d 1h after the Car injection the value had increased significantly in the Sham-HP, antibody and inhibitor groups compared with the HP group, while the expressions of IL-33, ST2 and TNF-α had decreased significantly. In experiment two, by 4 hours after the Car injection, the average MPT of all the other groups had decreased significantly compared with the Sham-HP group. Moreover, by 7d 1h after the Car injection, the average MPTs of the groups receiving TENS had increased significantly, with significantly lower MPT in the TENS Ⅱ group than in group Ⅰ, on average. There was also significantly higher expression of IL-33, ST2 and TNF-α in group II. Compared with the TENS Ⅰ and Ⅱ groups, the average MPT was significantly higher in the TENS I and Ⅱ inhibitor groups, but IL-33, ST2 and TNF-α expression was lower.Conclusions:TENS can inhibit TNF-α expression, which influences the signals of the DRG IL-33/ST2 signaling pathway to reverse hyperalgesia. TENS combined with anti-TNF-α treatment is superior to TENS alone in treating hyperalgesia.

OBJECTIVE: To explore the feasibility and accuracy of ultrasound volume navigation (UVN) combined with X-ray fluoroscopy-guided percutaneous pedicle screw implantation through a prospective randomized controlled study. METHODS: Patients with thoracic and lumbar vertebral fractures scheduled for percutaneous pedicle screw fixation between January 2022 and January 2023 were enrolled. Among them, 60 patients met the selection criteria and were included in the study. There were 28 males and 32 females, with an average age of 49.5 years (range, 29-60 years). The cause of injury included 20 cases of traffic accidents, 21 cases of falls, 17 cases of slips, and 2 cases of heavy object impact. The interval from injury to hospital admission ranged from 1 to 5 days (mean, 1.57 days). The fracture located at T ₁₂ in 15 cases, L ₁ in 20 cases, L ₂ in 19 cases, and L ₃ in 6 cases. The study used each patient as their own control, randomly guiding pedicle screw implantation using UVN combined with X-ray fluoroscopy on one side of the vertebral body and the adjacent segment (trial group), while the other side was implanted under X-ray fluoroscopy (control group). A total of 4 screws and 2 rods were implanted in each patient. The implantation time and fluoroscopy frequency during implantation of each screw, angle deviation and distance deviation between actual and preoperative planned trajectory by imaging examination, and the occurrence of zygapophysial joint invasion were recorded. RESULTS: In terms of screw implantation time, fluoroscopy frequency, angle deviation, distance deviation, and incidence of zygapophysial joint invasion, the trial group showed superior results compared to the control group, and the differences were significant ( P<0.05). CONCLUSION UVN combined with X-ray fluoroscopy-guided percutaneous pedicle screw implantation can yreduce screw implantation time, adjust dynamically, reduce operational difficulty, and reduce radiation damage.
Male ; Female ; Humans ; Middle Aged ; Pedicle Screws ; Prospective Studies ; X-Rays ; Surgery, Computer-Assisted/methods* ; Spinal Fusion/methods* ; Fluoroscopy/methods* ; Lumbar Vertebrae/injuries*

Mitochondrial DNA (mtDNA) mutations result in a variety of genetic diseases. As an emerging therapeutic method, mtDNA editing technology recognizes targets more based on the protein and less on the nucleic acid. Although the protein recognition type mtDNA editing technology represented by zinc finger nuclease technology, transcription activator like effector nuclease technology and base editing technology has made some progress, the disadvantages of complex recognition sequence design hinder further popularization. Gene editing based on nucleic acid recognition by the CRISPR system shows superiority due to the simple structure, easy design and modification. However, the lack of effective means to deliver nucleic acids into mitochondria limits application in the field of mtDNA editing. With the advances in the study of endogenous and exogenous import pathways and the deepening understanding of DNA repair mechanisms, growing evidence shows the feasibility of nucleic acid delivery and the broad application prospects of nucleic acid recognition type mtDNA editing technology. Based on the classification of recognition elements, this article summarizes the current principles and development of mitochondrial gene editing technology, and discusses its application prospects.
Genes, Mitochondrial ; Gene Editing ; Mitochondria/genetics* ; DNA, Mitochondrial/genetics* ; Nucleic Acids ; Technology

OBJECTIVE: To explore the anti-inflammatory effects of ethyl lithospermate in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine-derived macrophages and zebrafish, and its underlying mechanisms. METHODS: 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide (MTT) assays were performed to investigate the toxicity of ethyl lithospermate at different concentrations (12.5-100 µ mol/L) in RAW 264.7 cells. The cells were stimulated with LPS (100 ng/mL) for 12 h to establish an inflammation model in vitro, the production of pro-inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor α (TNF-α) were assessed by enzyme linked immunosorbent assay (ELISA). Western blot was used to ascertain the protein expressions of signal transducer and activator of transcription 3 (STAT3), nuclear factor kappa B (NF-κB) p65, phospho-STAT3 (p-STAT3, Tyr705), inhibitor of NF-κB (IκB) α, and phospho-I κB α (p-IκB α, Ser32), and confocal imaging was used to identify the nuclear translocation of NF-κB p65 and p-STAT3 (Tyr705). Additionally, the yolk sacs of zebrafish (3 days post fertilization) were injected with 2 nL LPS (0.5 mg/mL) to induce an inflammation model in vivo. Survival analysis, hematoxylin-eosin (HE) staining, observation of neutrophil migration, and quantitative real-time polymerase chain reaction (qRT-PCR) were used to further study the anti-inflammatory effects of ethyl lithospermate and its probable mechanisms in vivo. RESULTS: The non-toxic concentrations of ethyl lithospermate have been found to range from 12.5 to 100 µ mol/L. Ethyl lithospermate inhibited the release of IL-6 and TNF-α(P<0.05 or P<0.01), decreased IκBα degradation and phosphorylation (P<0.05) as well as the nuclear translocation of NF-κB p65 and p-STAT3 (Tyr705) in LPS-induced RAW 264.7 cells (P<0.01). Ethyl lithospermate also decreased inflammatory cells infiltration and neutrophil migration while increasing the survival rate of LPS-stimulated zebrafish (P<0.05 or P<0.01). In addition, ethyl lithospermate also inhibited the mRNA expression levels of of IL-6, TNF-α, IκBα, STAT3, and NF-κB in LPS-stimulated zebrafish (P<0.01). CONCLUSION Ethyl lithospermate exerts anti-Inflammatory effected by inhibiting the NF-κB and STAT3 signal pathways in RAW 264.7 macrophages and zebrafish.
Animals ; Mice ; NF-kappa B/metabolism* ; Lipopolysaccharides ; RAW 264.7 Cells ; Zebrafish ; NF-KappaB Inhibitor alpha/metabolism* ; Interleukin-6/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; STAT3 Transcription Factor/metabolism* ; Inflammation/metabolism* ; Anti-Inflammatory Agents/therapeutic use*

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15％ of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.T260A, p.R422W, and p.R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5％?49.7％ of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3％?17.9％, which was significantly higher than that (6.9％?7.4％) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.‍T260A, p.‍R422W, and p.‍R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‍‒‍49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‍‒‍17.9%, which was significantly higher than that (6.9%‍‒‍7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
Humans ; Apoptosis Inducing Factor/metabolism* ; NAD/metabolism* ; Dimerization ; Apoptosis

Objective:To analyze the case fatality rate of HIV/AIDS cases and influencing factors in Jingzhou.Methods:The data were retrieved from HIV/AIDS Comprehensive Response Information System and the cases diagnosed with HIV/AIDS in Jingzhou during 1996-2021 and aged 15 years or older were selected for the study. The death curve was drawn with Kaplan-Meier method, and Cox proportional-hazards model was used to identify influencing factors for death.Results:A total of 3 304 HIV/AIDS cases were followed up for 16 091.5 person-years, and 893 cases died, with a case fatality rate of 5.5/100 person-years. The cumulative case fatality rates of 1, 5 and 10 years were 15.4%, 25.0% and 34.6% respectively, the cumulative case fatality rates of 1, 5 and 10 years were 6.9%, 14.4% and 23.7% in the cases with access to antiretroviral therapy (ART), and 68.0%, 90.1% and 98.7% in the cases without access to ART. The results of Cox proportional hazards regression model showed that the risk for death was higher in those without access to ART than in those with access to ART (a HR=9.85, 95% CI: 8.19-11.85). The risk factors for death in those with access to ART included being men (a HR=1.64, 95% CI: 1.29-2.08), age ≥60 years old at diagnosis (a HR=3.52, 95% CI: 2.38-5.20), being infected by injecting drug use/others (a HR=2.38, 95% CI:1.30-4.34), being detected by medical institution (a HR=1.53, 95% CI: 1.11-2.11), CD4 +T lymphocytes(CD4) counts <50 cells/μl (a HR=2.58, 95% CI: 1.87-3.58). The protective factor for death was high education level (high school and technical secondary school: a HR=0.64,95% CI:0.46-0.90; college and above: a HR=0.42, 95% CI: 0.24-0.73). The risk factors for HIV/AIDS death in those without access to ART included older age at diagnosis (30-44 years old: a HR=2.32, 95% CI: 1.40-3.84; 45-59 years old:a HR=2.61, 95% CI: 1.59-4.27; ≥60 years old: a HR=3.31, 95% CI: 2.01-5.47), lower CD4 counts (<50 cells/μl: a HR=10.47, 95% CI: 6.47-16.56; 50-199 cells/μl: a HR=2.31, 95% CI: 1.08-4.94; 200-349 cells/μl: a HR=2.35, 95% CI: 1.46-3.79). Conclusions:The case fatality rate of HIV/AIDS was relatively high in Jingzhou from 1996 to 2021, the first CD4 counts, ART and age at diagnosis were the major factors affecting HIV/AIDS death, "Expanding testing" and "prompt treatment upon diagnosis" should be continued and enhanced to improve the efficacy of ART and HIV/AIDS case survival.

Background Macrophages are essential components of the natural immune system. They play a significant role in resisting foreign bodies in the respiratory tract and maintaining the homeostasis of the internal environment of lung tissue. Objective To investigate the mechanism of macrophage pyroptosis induced by silica dust with different particle sizes. Methods The modified murine macrophage cell line, RAW-ASC cells, was cultured and divided into a blank control group, a lipopolysaccharide (LPS) group (1 μg·mL⁻¹ LPS), a nano-SiO₂ group (1 μg·mL⁻¹ LPS+100 μg·mL⁻¹ nano-SiO₂), a micro-SiO₂ group (1 μg·mL⁻¹ LPS+750 μg·mL⁻¹ micro-SiO₂), and a positive control group [1 μg·mL⁻¹ LPS+3 mmol·L⁻¹ adenosine triphosphate (ATP)]. Apart from the blank control group, cells in other groups were pretreated with LPS for 6 h, and then exposed to SiO₂ or ATP for 4 h. According to the molecular target NOD-like receptor pyrin domain-containing protein 3 (NLRP3) and reactive oxygen species (ROS), we applied MCC950 (NLRP3 inhibitor) and N-acetyl cysteine (NAC, ROS scavenger) to macrophages. CCK-8 assay was used to detect cell viability; 5-ethynyl-2'-deoxyuridine (EdU) staining was used to detect cell proliferation; lactate dehydrogenase (LDH) assay kit was used to detect LDH in supernatant; calcein AM/PI fluorescent double-staining was applied to evaluate cell rupture; 2',7'-dichlorofluorescin diacetate (DCFH-DA) fluorescent probe was used to measure the content of ROS; Western blotting was used to measure the expressions of NLRP3, apoptosis-associated speck-like protein containing CARD (ASC), Caspase-1, gasdermin D (GSDMD), and interleukin-1β (IL-1β). Results Compared with the blank group, 100 μg·mL^-1nano-SiO₂ and 750 μg·mL^-1micro-SiO₂ dust exposure reduced the cell viability to 40% and 68% (P<0.05), and the cell proliferation rate to 30% and 33% (P<0.01), respectively; they also induced cell lysis and ROS release, upregulated NLRP3, ASC, Caspase-1, GSDMD, and IL-1β at protein level (P<0.05), and induced macrophage pyroptosis. After intervening with MCC950 (10 μmol·L^-1) and NAC (10 mmol·L^-1), the expressions of NLRP3, ASC, Caspase-1, and IL-1β decreased (P<0.05), and, specifically, NAC effectively reduced ROS levels (P<0.05). Conclusion Both nano- and micro-SiO₂ dust have cytotoxicity, can upregulate ROS level, activate NLRP3 inflammasome, and promote the release of cytokines, leading to pyroptosis. These results are helpful to reveal the molecular mechanism of macrophage pyroptosis induced by SiO₂ dust.

Objective:To retrospectively analyze the setup errors of thermoplastic head and shoulder molds alone or combined with vacuum pad in hypofractionated stereotactic radiotherapy (HFSRT) for non-small cell lung cancer (NSCLC) with brain metastases.Methods:Fifty-four NSCLC patients with brain metastases who received HFSRT from 2017 to 2019 were enrolled in this study. Twenty-four patients were fixed with thermoplastic head and shoulder molds (group A), and 30 patients were fixed with thermoplastic head and shoulder molds plus vacuum pad (group B). The interfraction and intrafraction setup errors were acquired from cone-beam CT online image registration before and after the HFSRT. Optical surface system was applied in monitoring the intrafraction setup errors. The setup errors in each direction between two groups were analyzed by independent samples t-test. Results:For the interfraction setup errors of the whole group, the proportion of the horizontal setup errors of ≥3mm was 7.0% to 15.4% and 7.0% to 12.6% for the rotation setup errors of ≥2°. In group A, the anteroposterior setup error was (1.035±1.180)mm, significantly less than (1.512±0.955)mm in group B ( P=0.009). In group A, the sagittal rotation setup error was 0.665°±0.582°, significantly less than 0.921°±0.682° in group B ( P=0.021). For the intrafraction setup errors of the whole group, the proportion of horizontal setup errors of ≥1mm was 0% to 0.7%, whereas no rotation setup error of ≥1° were observed. In group B, bilateral, anteroposterior and sagittal rotation setup errors were (0.047±0.212)mm, (0.023±0.152)mm and 0.091°±0.090°, significantly less compared with (0.246±0.474)mm, (0.140±0.350)mm and 0.181°±0.210° in group A ( P=0.004, P=0.020, P=0.001), respectively. Optical surface monitoring data were consistent with the obtained results. Conclusions:Thermoplastic head and shoulder molds (with or without vacuum pad) combined with online image registration and six-dimensional robotic couch correction can be applied in HFSRT for brain metastases from NSCLC. The intrafraction setup errors in group B are smaller than those in group A. Optical surface system has certain value in monitoring the intrafractional movement.

Objective:To explore the application of the Simodont dental trainer in preclinical manual dexterity training for dental students of different grades, and to discuss its effect by quantitatively evaluating their practice results.Methods:The evaluation was conducted among 118 students in the Department of Stomatology, Shantou University Medical College from Batch 2015 to Batch 2018. Each student had five different manual dexterity modules for training. Each module had three levels of difficulty and was required to be completed in 30 minutes. The assessment index included Target, Leeway Bottom and Sides, Container Bottom and Sides, which was provided by the Simodont dental trainer. The operating time (in seconds), the displacement of the dental hand pieces and the dental mirrors (in meters) were simultaneously recorded. SPSS 25.0 was used for statistical analysis.Results:For the index Target, Leeway Bottom and Sides, Container Bottom and Sides, the results showed that there was a statistical difference between students who passed pre-clinical training and who did not take the professional course ( P < 0.001). For the index Target, differences were showed among students from different grades in all modules ( P < 0.001). Conclusion:The circle module, channel module, hollow-circle module and cross-module in the Simodont dental trainer have sensitivity to discriminate the manual dexterity of different levels of dental students. The further assessment of the discrimination of the manual dexterity is required for assume-block module. The Simodont dental trainer can quantitatively measure the manual dexterity of dental students, which is important for the quantitative evaluation of dental preclinical education.