Clinical study on freehand of bicortical sacral screw fixation with the assistance of torque measurement device.

BACKGROUND: Sacral screw loosening is a typical complication after internal fixation surgery through the vertebral arch system. Bicortical fixation can successfully prevent screw loosening, and how improving the rate of bicortical fixation is a challenging clinical investigation. OBJECTIVE: To investigate the feasibility of improving the double corticality of sacral screws and the optimal fixation depth to achieve double cortical fixation by combining the torque measurement method with bare hands. METHODS: Ninety-seven cases of posterior lumbar internal fixation with pedicle root system were included in this study. Based on the tactile feedback of the surgeon indicating the expected penetration of the screw into the contralateral cortex of the sacrum, the screws were further rotated by 180°, 360°, or 720°, categorized into the bicortical 180° group, bicortical 360° group, and bicortical 720° group, respectively. Intraoperatively, the torque during screw insertion was recorded. Postoperatively, the rate of double-cortex engagement was evaluated at 7 days, and screw loosening was assessed at 1 year follow-up. RESULTS: The bicortical rates of the 180° group, 360° group, and 720° group were 66.13%, 91.18% and 93.75%, respectively. There were statistically significant differences between the 180° group and both the 360° and 720° groups (P < 0.05). However, there was no statistically significant difference between the 360° group and the 720° group (P > 0.05).The rates of loosening of sacral screws in the 180° group, 360° group, and 720° group were 20.97%, 7.35% and 7.81%, respectively. There were statistically significant differences between the 180° group and both the 360° and 720° groups (P < 0.05). However, there was no statistically significant difference between the 360° group and the 720° group (P > 0.05). The bicortical 360° group achieved a relatively satisfactory rate of dual cortical purchase while maintaining a lower rate of screw loosening. CONCLUSION: Manual insertion of sacral screws with the assistance of a torque measurement device can achieve a relatively satisfactory dual cortical purchase rate while reducing patient hospitalization costs.

Prediction of Screw Loosening After Dynamic Pedicle Screw Fixation With Lumbar Polyetheretherketone Rods Using Magnetic Resonance Imaging-Based Vertebral Bone Quality Score.

OBJECTIVE: To investigate the correlation between magnetic resonance imaging-based vertebral bone quality (VBQ) score and screw loosening after dynamic pedicle screw fixation with polyetheretherketone (PEEK) rods, and evaluate its predictive value. METHODS: A retrospective analysis was conducted on the patients who underwent dynamic pedicle screw fixation with PEEK rods from March 2017 to June 2022. Data on age, sex, body mass index, hypertension, diabetes, hyperlipidemia history, long-term smoking, alcohol consumption, VBQ score, L1-4 average Hounsfield unit (HU) value, surgical fixation length, and the lowest instrumented vertebra were collected. Logistic regression analysis was employed to assess the relationship between VBQ score and pedicle screw loosening (PSL). RESULTS: A total of 24 patients experienced PSL after surgery (20.5%). PSL group and non-PSL group showed statistical differences in age, number of fixed segments, fixation to the sacrum, L1-4 average HU value, and VBQ score (p < 0.05). The VBQ score in the PSL group was higher than that in the non-PSL group (3.56 ± 0.45 vs. 2.77 ± 0.31, p < 0.001). In logistic regression analysis, VBQ score (odds ratio, 3.425; 95% confidence interval, 1.552-8.279) were identified as independent risk factors for screw loosening. The area under the receiver operating characteristic curve for VBQ score predicting PSL was 0.819 (p < 0.05), with the optimal threshold of 3.15 (sensitivity, 83.1%; specificity, 80.5%). CONCLUSION: The VBQ score can independently predict postoperative screw loosening in patients undergoing lumbar dynamic pedicle screw fixation with PEEK rods, and its predictive value is comparable to HU value.

Correlation between axial symptoms and cervical sagittal alignment parameters in patients with two-level or three-level cervical spondylotic myelopathy: anterior cervcial discectomy and fusion versus hybird surgery.

OBJECTIVE: To comparatively analyze the correlation between axial symptoms (AS) and cervical sagittal alignment parameters after anterior cervical discectomy and fusion (ACDF) and hybrid surgery (HS). METHODS: From January 2018 to June 2023, 74 patients who underwent ACDF (n = 36) or HS (n = 38) for two-level or three-level cervical spondylotic myelopathy were retrospectively analyzed. The Visual Analogue Scale (VAS), Japanese Orthopedic Association (JOA), Neck Disability Index (NDI) were recorded to assess clinical outcomes. Cervical sagittal alignment parameters (Cobb's angle C2-7, C7 slope [C7S], and C2-7 sagittal vertical axis [C2-7 SVA]) were measured preoperatively, 3 days postoperatively, and at the last follow-up. The range of motion (ROM) of C2-7 and ROM of surgical segment were measured. The occurrence of AS was observed at the last follow-up. Logistic regression was used to analyze the correlation between postoperative AS and cervical sagittal alignment parameters. RESULTS: Both in ACDF group and HS group, VAS, JOA and NDI scores showed significant improvements at 3-day postoperation and at the last follow-up (P < 0.05). However, there was no significant difference between the two groups (P > 0.05). The Cobb's angle C2-7 and C7S were significantly increased at 3 days postoperation compared with pre-operatively in both groups (P < 0.05). C2-7SVA was increased in both groups 3 days after surgery compared with pre-operatively, but there was no significant difference (P > 0.05). At the last follow-up, the ROM of C2-7 in ACDF group was significantly smaller than HS group (P < 0.05). The prevalence of postoperative AS in the ACDF group and HS group was 41.7 and 18.4%, respectively, with statistical difference between the two groups (P < 0.05). When simple Logistic regression analysis was used, the last Cobb's angle C2-7 (ß = -0.088), the last C2-7SVA (ß = 0.099) in ACDF group and the last C2-7SVA (ß = 0.222) in HS group were all correlated with the occurrence of postoperative AS. When multiple Logistic regression analysis was used, only the last C2-7SVA (ß = 0.181) in the HS group was positively correlated with the occurrence of postoperative AS. CONCLUSIONS: Both ACDF and HS can achieve satisfied clinical outcomes. ACDF and HS can improve cervical sagittal balance to a certain extent, and HS is superior to ACDF in maintaining ROM. The decrease of the last Cobb's angle C2-7 and the increase of the last C2-7SVA may be related to the occurrence of AS after ACDF. The increase of the last C2-7SVA was an independent risk factor for the occurrence of AS after HS.

The effects of ultrasonic processing on the antioxidant activity of Geotrichum candidum LG-8 and its cell wall extracts.

Introduction: Extraction techniques that influence cell wall polysaccharides (EPS) is crucial for maximizing their bioactivity. This study evaluates ultrasound technology for extracting antioxidant polysaccharides from Geotrichum candidum LG-8, assessing its impacton antioxidant activity. Methods: Ultrasound extraction of EPS from G. candidum LG-8 was optimized (18 min, pH 7.0, 40 W/cm2, 0.75 M NaCl). ABTS scavenging efficiency and monosaccharide composition of LG-EPS1 and LG-EPS3 were analyzed using Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). Results: The Results showed that ultrasonic treatment markedly increased the ABTS radical scavenging efficiency of LG-8 cells by 47%. At a concentration of 1 mg/mL, the ultrasonically extracted LG-EPS1 and LG-EPS3 polysaccharides exhibited significant ABTS radical scavenging efficiencies of 26% and 51%, respectively. Monosaccharide composition analysis identified mannose and glucose in LG-EPS1, while LG-EPS3 was primarily composed of mannose. FTIR spectra verified the polysaccharides' presence, and SEM provided visual confirmation of the nanoparticle structures characteristic of LG-EPS1 and LG-EPS3. Discussion: This research not only underscores the technological merits of ultrasound in polysaccharide extraction but also highlights the potential of G. candidum LG-8 derived polysaccharides as valuable bioactive compounds for antioxidant utilization.

The application of stem cell sheets for neuronal regeneration after spinal cord injury: a systematic review of pre-clinical studies.

BACKGROUND: Stem cell sheet implantation offers a promising avenue for spinal cord injury (SCI) and is currently under investigation in pre-clinical in vivo studies. Nevertheless, a systematic review of the relevant literature is yet to be performed. Thus, this systematic review aims to explore the efficacy of stem cell sheet technology in treating SCI, as indicated by experimental animal model studies. METHODS: We searched PubMed, EMBASE, and Web of Science. Manuscripts that did not pertain to in vivo pre-clinical studies and those published in non-English languages were excluded. A risk assessment for bias was performed using the SYRCLE tool. Extracted data were synthesized only qualitatively because the data were not suitable for conducting the meta-analysis. RESULTS: Among the 847 studies retrieved from electronic database searches, seven met the inclusion criteria. Six of these studies employed a complete transection model, while one utilized a compression model. Stem cell sources included bone marrow mesenchymal stem cells, stem cells from human exfoliated deciduous teeth, and adipose-derived mesenchymal stem cells. In all included studies, stem cell sheet application significantly improved motor and sensory functional scores compared to intreated SCI rats. This functional recovery correlated with histological improvements at the injury site. All studies are at low risk of bias but certain domains were not reported by some or all of the studies. CONCLUSION: The results of our systematic review suggest that stem cell sheets may be a feasible therapeutic approach for the treatment of SCI. Future research should be conducted on stem cell sheets in various animal models and types of SCI, and careful validation is necessary before translating stem cell sheets into clinical studies.

Biomechanical and clinical research of Isobar semi-rigid stabilization devices for lumbar degenerative diseases: a systematic review.

While lumbar spinal fusion using rigid rods is a prevalent surgical technique, it can lead to complications such as adjacent segment disease (ASDis). Dynamic stabilization devices serve to maintain physiological spinal motion and alleviate painful stress, yet they are accompanied by a substantial incidence of construct failure and subsequent reoperation. Compared to traditional rigid devices, Isobar TTL semi-rigid stabilization devices demonstrate equivalent stiffness and effective stabilization capabilities. Furthermore, when contrasted with dynamic stabilization techniques, semi-rigid stabilization offers improved load distribution, a broader range of motion within the fixed segment, and reduced mechanical failure rates. This paper will review and evaluate the clinical and biomechanical performance of Isobar TTL semi-rigid stabilization devices. A literature search using the PubMed, EMBASE, CNKI, Wanfang, VIP, and Cochrane Library databases identified studies that met the eligibility criteria. Twenty-eight clinical studies and nine biomechanical studies were included in this systematic review. The VAS, the ODI, and Japanese Orthopedic Association scoring improved significantly in most studies. UCLA grading scale, Pfirrmann grading, and modified Pfirrmann grading of the upper adjacent segments improved significantly in most studies. The occurrence rate of ASD was low. In biomechanical studies, Isobar TTL demonstrated a superior load sharing distribution, a larger fixed segment range of motion, and reduced stress at the rod-screw/screw-bone interfaces compared with titanium rods. While findings from mechanical studies provided promising results, the clinical studies exhibited low methodological quality. As a result, the available evidence does not possess sufficient strength to substantiate superior outcomes with Isobar semi-rigid system in comparison to titanium rods. To establish more conclusive conclusions, further investigations incorporating improved protocols, larger sample sizes, and extended follow-up durations are warranted.

Isobar hybrid dynamic stabilization with posterolateral fusion in mild and moderate lumbar degenerative disease.

OBJECTIVE: The aim of this study was to investigate the feasibility of using the Isobar TTL system and posterolateral fusion in a two-segment hybrid fixation approach, combined with spinal decompression, for treating mild and moderate lumbar degenerative disease. Specifically, we sought to evaluate the effectiveness of this approach for managing two-segment mild and moderate lumbar degenerative disease, and to determine whether it could provide a safe and reliable alternative to traditional surgical methods. METHODS: This retrospective study included 45 consecutive patients with two-level lumbar disc herniation or spinal stenosis, 24 of whom underwent the TTL system and posterolateral fusion combined (TTL group), and 21 of whom underwent posterolateral fusion alone (Rigid group). The surgical segment, admission diagnosis, operation time, and intraoperative bleeding were recorded separately for the two groups of patients. Imaging studies included pre- and postoperative radiography, magnetic resonance imaging, and computed tomography. The clinical outcomes were measured by Oswestry Disability Index (ODI) scores, and a visual analogue scale (VAS) for back and leg pain. RESULTS: All patients completed the surgery successfully with a mean follow-up of 56.09 months. The operative time and intraoperative bleeding were lower in the TTL group than in the Rigid group (p < 0.05). All patients showed significant improvements in clinical outcomes, including VAS for back and leg pain, and ODI scores (p < 0.05). ODI scores, the TTL group was better than the Rigid group at 1 year after surgery and at the final follow-up (p < 0.05). Postoperative surgical segment range of motion (ROM) decreased in both groups (p < 0.05). The postoperative ROM of the upper adjacent segment increased in both groups and was significantly higher in both groups at the last follow-up compared with the preoperative period (p < 0.05), and the superior adjacent segment rom of the TTL group was lower than the Rigid group (p < 0.05). The modified Pfrrmann classification of the superior adjacent segment was significantly increased in both groups at the last follow-up (p < 0.05). And in the TTL group, ROM, DH, and modified Pfrrmann grading of dynamic segment outperformed fusion segments. According to the UCLA classification, the incidence of adjacent segment degeneration (ASD) was 4.2% in the TTL group and 23.8% in the Rigid group, and the incidence of ASD was lower in the TTL group than in the Rigid group (P < 0.05). CONCLUSION: The Isobar TTL System was utilized in two-level lumbar hybrid surgery, resulting in no evident indications of lumbar instability being detected on X-rays captured at a minimum of 4 years after the operation, while retaining partial range of motion of the surgical segment. The general clinical efficacy is equivalent to titanium rod fusion surgery, presenting an alternative treatment for individuals with mild and moderate lumbar degenerative disease.

Correlation between surgical segment mobility and paravertebral muscle fatty infiltration of upper adjacent segment in single-segment LDD patients: retrospective study at a minimum 2 years' follow-up.

OBJECTIVE: The purpose of this study was to investigate the relationship between surgical segment mobility and fatty infiltration of the adjacent segment paravertebral muscles in patients with single-segment lumbar degenerative disease (LDD) who underwent decompression with fusion or dynamic stabilization. METHODS: Retrospective analysis of patients who underwent lumbar decompression combined with titanium rod fixation intertransverse fusion (PITF group), Isobar TTL dynamic stabilization (TTL group) or Isobar EVO dynamic stabilization (EVO group) for single-segment lumbar degenerative disease, from March 2012 to July 2018. The preoperative and final follow-up clinical indexes C-LDSI and the measured imaging indexes (range of motion of the surgical segment and the upper adjacent segment, and Goutallier grade of the upper adjacent segment) were counted, and the differences between the preoperative and final follow-up indexes were compared. RESULTS: According to the inclusion and exclusion criteria, 68 patients were included in this study, 21 in the PITF group, 24 in the TTL group, and 23 in the EVO group. At the final follow-up, the C-LSDI score had significantly higher in the PITF group than the TTL and EVO groups, and the C-LSDI score was a very strongly negatively correlated with ROM of surgical segment (r=-0.7968, p < 0.001). There was a strong negative correlation between surgical segment and upper adjacent segment mobility (r = -0.6959, p < 0.001). And there was a very strong negative correlation between ROM of surgical segment and upper adjacent segment paravertebral muscle Goutallier classification (r = -0.8092, p < 0.001), whereas the ROM of the upper adjacent segment was strong positive correlated with the Goutallier classification (r = 0.6703, P < 0.001). CONCLUSION: Compared with decompression combined with rigid fusion, decompression combined with dynamic fixation for single-segment lumbar degenerative disease can significantly reduce postoperative low back stiffness. And a certain range of increased mobility of the dynamic stabilization device can effectively reduce the compensatory mobility of the upper adjacent segment and slow down the fatty infiltration of the paravertebral muscle in the adjacent segment.

Capacity and kinetics of zearalenone adsorption by Geotrichum candidum LG-8 and its dried fragments in solution.

The application of LG-8 and its dry fragments as zearalenone (ZEN) adsorbents was investigated. The study showed that Geotrichum candidum LG-8 and its fragments dried at 55°C or through lyophilization are able to adsorb around 80% of ZEN. However, besides in water and 55°C-drying conditions, SEM indicated that higher 90% of ZEN binding tended to occur when cell walls of fragments were intact with less adhesion among themselves. Notably, ZEN/LG-8 fragments complexes were quite stable, as only 1.262% and 1.969% of ZEN were released after successive pH treatments for 4 h and 5 min. The kinetic data signified that adsorption of ZEN onto LG-8 fragments followed well the pseudo-first-order kinetic model. Isotherm calculations showed Langmuir model was favourable and monolayer adsorption of ZEN occurred at functional binding sites on fragments surface. Therefore, we conclude that it can be an alternative biosorbent to treat water contained with ZEN, since LG-8 is low-cost biomass and its fragments have a considerable high biosorption capacity avoiding impacting final product quality and immunodeficient patients.

Comparison between single-segment Isobar EVO dynamic stabilization and Isobar TTL dynamic stabilization in the treatment of lumbar degenerative diseases: a single center retrospective study over 4 years.

OBJECTIVE: Posterior instrumented fusion is the most widely accepted surgical treatment for spinal stenosis and disc herniation. However, fusion can affect daily activities due to lumbar stiffness. In recent years, dynamic stabilization has been introduced to overcome the drawbacks of fusion, however, it is inconclusive whether dynamic stabilization requires the maintenance of a level of activity that is closer to the physiological state of activity for better clinical efficacy. The purpose of this study was to compare the effectiveness of dynamic stabilization with different levels of activity (Isobar EVO and TTL) in the treatment of spinal stenosis and disc herniation. METHODS: This study retrospectively reviewed 80 consecutive patients with lumbar degenerative diseases who were undergoing surgical treatment between March 2014 and July 2018. 41 patients (EVO group) and 39 patients (TTL group) underwent fenestrated decompression with Isobar EVO stabilization and Isobar TTL stabilization, respectively. Clinical outcomes, radiographic data, and postoperative complications were compared between the two groups. RESULTS: At an average follow-up of 52.23 ± 2.97 months, there were no significant differences in the oswestry disability index (ODI) (P > 0.05). The visual analog scale for back pain (VASback) and visual analog scale for the leg pain (VASleg) of the EVO group were lower compared with the TTL group (P < 0.05). The range of motion (ROM) of operated segments were significantly higher in the EVO group as compared to the TTL group (P < 0.05). The intervertebral space height (ISH) of upper adjacent segments were significantly higher in the EVO group as compared to the TTL group (P < 0.05). The overall complications were less in the EVO group, but the difference was not statistically significant (P > 0.05). CONCLUSION: Both Isobar EVO dynamic stabilization and TTL dynamic stabilization can improve clinical outcomes of patients with spinal stenosis and disc herniation. Isobar EVO has advantages over Isobar TTL in terms of improving low back and leg pain, maintaining mobility of the operated segment, and preventing further degeneration of the upper adjacent segment.

Effects of posterior lumbar nonfusion surgery with isobar devices versus posterior lumbar interbody fusion surgery on clinical and radiological features in patients with lumbar degenerative diseases: a meta-analysis.

PURPOSE: The aim of this study was to systematically evaluate the efficacy of posterior lumbar isobar nonfusion with isobar devices versus posterior lumbar interbody fusion (PLIF) in the treatment of patients with lumbar degenerative diseases (LDDs). MATERIALS AND METHOD: We performed a literature review and meta-analysis in accordance with the Cochrane methodology. The analysis included a Group Reading Assessment and Diagnostic Evaluation assessments, Jadad Quality Score evaluations, and Risk of Bias in Randomized Studies of Interventions assessments. The PubMed, Ovid, EMBASE, Web of Science, MEDLINE, CNKI, VIP and WanFang databases were searched to collect and compare relevant randomized controlled trials and cohort studies of isobar nonfusion and PLIF in the treatment of lumbar degenerative diseases. The retrieval time was from database inception to June 2021. Two evaluators independently screened the literature, extracted data, and evaluated the quality of the included studies. Outcome measures of interest included low back pain, disability, and radiological features. The protocol for this systematic review was registered on INPLASY (2021110059) and is available in full on inplasy.com ( https://inplasy.com/inplasy-2021-11-0059/ ). RESULTS: Of the 7 RCTs, 394 patients met the inclusion criteria. The meta-analysis results showed that isobar nonfusion surgery shortened the surgical duration (P = 0.03), reducing intraoperative bleeding (P = 0.001), retained the ROM of surgical segment (P < 0.00001) and the ROM of the lumbar spine (P < 0.00001), and reduced the incidence of ASD (P = 0.0001). However, no significant difference in the postoperative ODI index (P = 0.81), VAS score of LBP (P = 0.59, VAS score of lower limb pain (P = 0.05, and JOA score (P = 0.27) was noted. CONCLUSIONS: Posterior lumbar nonfusion surgery with isobar devices is superior to PLIF in shortening the surgical duration, reducing intraoperative bleeding, retaining the ROM of surgical segments and the lumbar spine to a certain extent, and preventing ASD. Given the possible publication bias, we recommend further large-scale studies.

Sanghuangporus toxicodendri sp. nov. (Hymenochaetales, Basidiomycota) from China.

Sanghuangporus toxicodendri (Hymenochaetales) is described as new based on collections made from Shennongjia Forestry District, Hubei Province, China. All studied basidiocarps grew on living trunks of Toxicodendron sp. This new species is characterized by having perennial, effused-reflexed to pileate basidiocarps; pore surface brownish yellow or yellowish brown, pores 7-9 per mm; context 1-5 mm thick or almost invisible; setae ventricose, dark brown, 26-42 × 7-10 µm; basidia 4-sterigmate or occasionally 2-sterigmate; basidiospores broadly ellipsoid, smooth, brownish yellow, slightly thick-walled, mostly 3.5-4 × 2.8-3 µm. Maximum likelihood and Bayesian inference phylogenies inferred from internal transcribed spacer (ITS) region of rDNA indicated that Sanghuangporus spp. formed a monophyletic clade and resolved as a sister to Tropicoporus spp., and six strains of S. toxicodendri formed a monophyletic group which is sister to S. quercicola. An identification key to known species of Sanghuangporus is provided.

Salidroside attenuates colistin-induced neurotoxicity in RSC96 Schwann cells through PI3K/Akt pathway.

Neurotoxicity is a key dose-limiting factor for colistin therapy. This study aimed to investigate the protective effect of Salidroside on colistin-induced neurotoxicity in RSC96 Schwann cells and the underlying mechanisms. After Salidroside (12.5, 25, 50 µg/mL) treatment for 2 h, the cells were cultured with 250 µg/mL colistin for 24 h. In order to investigate the role of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway, the cells were pre-treated with LY294002 (12.5 µmol/L, a specific inhibitor of PI3K phosphorylation) for 1 h before Salidroside (50 µg/mL) treatment, then were co-cultured with colistin (250 µg/mL) for 24 h. The results showed that colistin treatment could induce apoptotic cell death which was associated with oxidative stress injury. Salidroside could reduce colistin-induced neurotoxicity, decrease the effect of colistin on the reduced expression levels of p-Akt and Bcl-2, and increased the expresion of Bax, release of Cyt c, and activation of caspase-3. However, the protective effect of Salidroside against colistin-induced apoptosis was partly abolished by LY294002. These findings suggest that Salidroside could attenuate colistin-induced neurotoxicity in RSC96 Schwann cells via the PI3K/Akt pathway.

p53 Mediates Colistin-Induced Autophagy and Apoptosis in PC-12 Cells.

The mechanism of colistin-induced neurotoxicity is still unknown. Our recent study (L. Zhang, Y. H. Zhao, W. J. Ding, G. Z. Jiang, Z. Y. Lu, L. Li, J. L. Wang, J. Li, and J. C. Li, Antimicrob Agents Chemother 59:2189-2197, 2015, http://dx.doi.org/10.1128/AAC.04092-14; H. Jiang, J. C. Li, T. Zhou, C. H. Wang, H. Zhang, and H. Wang, Int J Mol Med 33:1298-1304, 2014, http://dx.doi.org/10.3892/ijmm.2014.1684) indicates that colistin induces autophagy and apoptosis in rat adrenal medulla PC-12 cells, and there is interplay between both cellular events. As an important cellular stress sensor, phosphoprotein p53 can trigger cell cycle arrest and apoptosis and regulate autophagy. The aim of the present study was to investigate the involvement of the p53 pathway in colistin-induced neurotoxicity in PC-12 cells. Specifically, cells were treated with colistin (125 µg/ml) in the absence and presence of a p53 inhibitor, pifithrin-α (PFT-α; 20 nM), for 12 h and 24 h, and the typical hallmarks of autophagy and apoptosis were examined by fluorescence/immunofluorescence microscopy and electron microscopy, real-time PCR, and Western blotting. The results indicate that colistin had a stimulatory effect on the expression levels of the target genes and proteins involved in autophagy and apoptosis, including LC3-II/I, p53, DRAM (damage-regulated autophagy modulator), PUMA (p53 upregulated modulator of apoptosis), Bax, p-AMPK (activated form of AMP-activated protein kinase), and caspase-3. In contrast, colistin appeared to have an inhibitory effect on the expression of p-mTOR (activated form of mammalian target of rapamycin), which is another target protein in autophagy. Importantly, analysis of the levels of p53 in the cells treated with colistin revealed an increase in nuclear p53 at 12 h and cytoplasmic p53 at 24 h. Pretreatment of colistin-treated cells with PFT-α inhibited autophagy and promoted colistin-induced apoptosis. This is the first study to demonstrate that colistin-induced autophagy and apoptosis are associated with the p53-mediated pathway.

Autophagy regulates colistin-induced apoptosis in PC-12 cells.

Colistin is a cyclic cationic polypeptide antibiotic with activity against multidrug-resistant Gram-negative bacteria. Our recent study demonstrated that colistin induces apoptosis in primary chick cortex neurons and PC-12 cells. Although apoptosis and autophagy have different impacts on cell fate, there is a complex interaction between them. Autophagy plays an important role as a homeostasis regulator by removing excessive or unnecessary proteins and damaged organelles. The aim of the present study was to investigate the modulation of autophagy and apoptosis regulation in PC-12 cells in response to colistin treatment. PC-12 cells were exposed to colistin (125 to 250 µg/ml), and autophagy was detected by visualization of monodansylcadaverine (MDC)-labeled vacuoles, LC3 (microtubule-associated protein 1 light chain 3) immunofluorescence microscopic examination, and Western blotting. Apoptosis was measured by flow cytometry, Hoechst 33258 staining, and Western blotting. Autophagosomes were observed after treatment with colistin for 12 h, and the levels of LC3-II gene expression were determined; observation and protein levels both indicated that colistin induced a high level of autophagy. Colistin treatment also led to apoptosis in PC-12 cells, and the level of caspase-3 expression increased over the 24-h period. Pretreatment of cells with 3-methyladenine (3-MA) increased colistin toxicity in PC-12 cells remarkably. However, rapamycin treatment significantly increased the expression levels of LC3-II and beclin 1 and decreased the rate of apoptosis of PC-12 cells. Our results demonstrate that colistin induced autophagy and apoptosis in PC-12 cells and that the latter was affected by the regulation of autophagy. It is very likely that autophagy plays a protective role in the reduction of colistin-induced cytotoxicity in neurons.

Protective effects of ginsenoside Rg1 against colistin sulfate-induced neurotoxicity in PC12 cells.

The present study aimed to examine the protective effect of ginsenoside Rg1 against colistin-induced neurotoxicity in cultured rat pheochromocytoma (PC12) cells. Ginsenoside Rg1 was shown to elevate cell viability, decrease levels of malondialdehyde and intracellular reactive oxygen species, enhance activity of superoxide dismutase and glutathione, and decrease the release of cytochrome-c, formation of DNA fragmentation in colistin-treated PC12 cells. Ginsenoside Rg1 also reversed the increased caspase-9 and -3 mRNA levels caused by colistin in PC12 cells. These results suggest that ginsenoside Rg1 exerts a neuroprotective effect on colistin-induced neurotoxicity in PC12 cells, at least in part, via the inhibition of oxidative stress, prevention of apoptosis mediated via mitochondria pathway. Co-administration of ginsenoside Rg1 highlights the potential to increase the therapeutic index of colistin.

Effects of Traditional Chinese Medicine Qinbai Qingfei Concentrated Pellet on Cellular Infectivity of Mycoplasma pneumoniae.

Aim. To study the effect and mechanism of traditional Chinese medicine Qinbai Qingfei concentrated pellet (QQCP) against Mycoplasma pneumoniae (MP). Methods. Rat airway smooth muscle (ASM) cells were used to examine the antimycoplasmal activity of QQCP via four drug-adding modes: pre- and postadding drugs, simultaneous-adding after drug and MP mixed, and simultaneous-adding drug and MP; taking roxithromycin dispersive tablets (RDT) as positive control, the cellular A 570 values were determined by MTT method. Results. All of A 570 values in QQCP group were significantly higher than those of the corresponding MP control group (P < 0.01) in four drug-adding modes; there was no significant difference in A 570 values between the QQCP group and that of the positive control group (P > 0.05), confirming that QQCP could significantly inhibit the infectivity of MP to ASM cells. Conclusion. QQCP had significant activity in preventing and treating MP infection, killing MP, and antiabsorption.