Recent advances in patient-derived tumor organoids for reconstructing TME of head and neck cancer.

BACKGROUND: The differences between existing preclinical models and the tumor microenvironment in vivo are one of the significant challenges hindering cancer therapy development. Patient-derived tumor organoids (PDTO) can highly retain tumor heterogeneity. Thus, it provides a more reliable platform for research in tumor biology, new drug screening, and precision medicine. METHODS: We conducted a systematic review to summarise the characteristics of the existing preclinical models, the advantages of patient-derived tumor organoids in reconstructing the tumor microenvironment, and the latest research progress. Moreover, this study deciphers organoid culture technology in the clinical precision treatment of head and neck cancer to achieve better transformation. Studies were identified through a comprehensive search of Ovid MEDLINE (Wolters Kluwer), PubMed (National Library of Medicine), web of Science (Thomson Reuters) and, Scopus (Elsevier) databases, without publication date or language restrictions. RESULTS: In tumor development, the interaction between cellular and non-cellular components in the tumor microenvironment (TME) has a crucial role. Co-culture, Air-liquid interface culture, microfluidics, and decellularized matrix have depicted great potential in reconstructing the tumor microenvironment and simulating tumor genesis, development, and metastasis. CONCLUSION: An accurate determination of stromal cells, immune cells, and extracellular matrix can be achieved by reconstructing the head and neck cancer tumor microenvironment using the PDTO model. Moreover, the interaction between head and neck cancer cells can also play an essential role in implementing the individualized precision treatment of head and neck cancer.

A case of fatal poisoning caused by etomidate: evidence from pathological and toxicological analyses.

Etomidate is a nonbarbiturate sedative derived from imidazole. Prolonged and excessive use of etomidate can lead to the suppression of adrenocortical function, myoclonus, and even death. This report describes a rare case of a 47-year-old man who died from acute intoxication after oral ingestion of liquid containing etomidate. The cause of death was conclusively attributed to etomidate based on a comprehensive investigation, including autopsy, histopathological examination, toxicological analysis, and biochemical analysis. This is the first reported case of a fatality solely resulting from the oral ingestion of etomidate, which can provide valuable insights for future forensic investigations involving etomidate poisoning. Therefore, it is imperative to share this case with the scientific community.

Gaps in forensic toxicological analysis: The veiled abrin.

Abrus precatorius is an herbaceous, flowering plant that is widely distributed in tropical and subtropical regions. Its toxic component, known as abrin, is classified as one of the potentially significant biological warfare agents and bioterrorism tools due to its high toxicity. Abrin poisoning can be utilized to cause accidents, suicides, and homicides, which necessitates attention from clinicians and forensic scientists. Although a few studies have recently identified the toxicological and pharmacological mechanisms of abrin, the exact mechanism remains unclear. Furthermore, the clinical symptoms and pathological changes induced by abrin poisoning have not been fully characterized, and there is a lack of standardized methods for identifying biological samples of the toxin. Therefore, there is an urgent need for further toxicopathologic studies and the development of detection methods for abrin in the field of forensic medicine. This review provides an overview of the clinical symptoms, pathological changes, metabolic changes, toxicologic mechanisms, and detection methods of abrin poisoning from the perspective of forensic toxicology. Additionally, the evidence on abrin in the field of forensic toxicology and forensic pathology is discussed. Overall, this review serves as a reference for understanding the toxicological mechanism of abrin, highlighting the clinical applications of the toxin, and aiding in the diagnosis and forensic identification of toxin poisoning.

Visualized sensing of erythritol using a simple enzyme-free catechol-based hydrogel film.

Based on the versatile properties of bio-derived materials, non-enzymatic assays in combination with electronic devices have attracted increasing interest. Here, we report a novel enzyme-free visualization approach for the detection of erythritol, which is a zero-calorie natural sweetener and serves as an ideal sucrose substitute for diabetics or overweight people who need sugar control. The recognition element of the electrochemical biosensor was constructed by catechol modification on a chitosan-based hydrogel film. The signal transduction was achieved by the competitive binding assay of sweeteners. The results show that 2-fluorophenylboronic acid (FPBA) can form a cyclic boronate ester with the ortho-hydroxyls of both reduced catechol and oxidized quinone, impeding the electron transfer and leading to redox signal attenuation. The addition of sweeteners caused a competitive reaction resulting in bonding between the 1,2-diols and FPBA moieties, and in the recovery of the redox signals. Importantly, the pattern of redox signal changes of catechol can be detected optically, as the oxidized quinone state is darker in color than the reduced catechol state. Using a simple cell phone imaging application, we demonstrate that erythritol can be distinguished from other sweeteners in real samples using the oxidized catechol-Chit0/agarose hydrogel film. Thus, we envision that this method could allow diabetics and people who need to control their sugar intake to detect whether the product contains only erythritol in the field or at home. In addition, this work further illustrates the potential of bio-derived materials for performing redox-based functions and enzyme-free visualization assays.

Application of nZnO supported with nanoclay for improving shrimp immunity.

This study discloses the nanoscale silicate platelet-supported nZnO (ZnONSP) applied as novel feed additives in aquaculture. The preparation of the nanohybrid (ZnO/NSP = 15/85, w/w) was characterized by UV-visible spectroscopy, powder X-ray diffraction and transmission electron microscope. The effects of ZnONSP on growth, zinc accumulation, stress response, immunity and resistance to Vibrio alginolyticus in white shrimp (Penaeus vannamei) were \demonstrated. To evaluate the safety of ZnONSP, shrimps (2.0 ± 0.3 g) were fed with ZnONSP containing diets (200, 400 and 800 mg/kg) for 56 days. Dietary ZnONSP did not affect the weight gain, specific growth rate, feed conversion ratio, survival rate, zinc accumulation, and the expression of heat shock protein 70 in tested shrimps. To examine the immunomodulatory effect of ZnONSP, shrimps (16.6 ± 2.4 g) were fed with the same experimental diets for 28 days. Dietary ZnONSP improved the immune responses of haemocyte in tested shrimps, including phagocytic rate, phagocytic index, respiratory burst, and phenoloxidase activity, and upregulated the expression of several genes, including lipopolysaccharide, ß-1,3-glucan binding protein, peroxinectin, penaeidin 2/3/4, lysozyme, crustin, anti-lipopolysaccharide factor, superoxide dismutase, glutathione peroxidase, clotting protein and α-2-macroglobulin. In the challenge experiment, shrimps (17.2 ± 1.8 g) were fed with ZnONSP containing diets (400 and 800 mg/kg) for 7 days and then infected with Vibrio alginolyticus. Notably, white shrimps that received ZnONSP (800 mg/kg) showed significantly improved Vibrio resistance, with a survival rate of 71.4 % at the end of 7-day observation. In conclusion, this study discovers that ZnONSP is a new type of immunomodulatory supplement that are effective on enhancing innate cellular and humoral immunities, and disease resistance in white shrimp.

In-situ synthesis of magnetic iron-chitosan-derived biochar as an efficient persulfate activator for phenol degradation.

Persulfate activation is a forceful method for eliminating organic pollutants from coal chemical wastewater. In this study, an in-situ synthesis method was used to fabricate an iron-chitosan-derived biochar (Fe-CS@BC) nanocomposite catalyst using chitosan as a template. Fe was successfully imprinted into the newly synthesized catalyst. The Fe-CS@BC can activate persulfate to effectively degrade phenol. This point was confirmed by scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The impact of various parameters on the removal rate was investigated in a single factor experiment. In Fe-CS@BC/PDS system, 95.96% of phenol (significantly higher than the original biochar of 34.33%) was removed within 45 min and 54.39% TOC within 2 h. The system showed superior efficiency over a broad pH value band from 3 to 9 and has a high degradation rate at ambient temperature. Free radical quenching experiment, EPR experiment and LSV experiment confirmed that multiple free radicals (including 1O2, SO4•-, O2•- and •OH) and electron transfer pathway combined to enhance phenol decomposition. Finally, the activation mechanism of persulfate by Fe-CS@BC was proposed to provide logical guidance on the treatment of organic pollutants in coal chemical wastewater.

Heat inactive Bacillus subtilis var. natto regulate Nile tilapia (Oreochromis niloticus) intestine microbiota and metabolites involved in the intestine phagosome response.

In this study, we evaluated the intestinal microbiota, intestinal and fecal metabolites production and the intestinal RNA-seq analysis of the Nile tilapia intestine after feeding with 105and 107 of the inactive Bacillus subtilis var. natto. First, we assessed the influence of heat inactive Bacillus subtilis var. natto on the growth performance, biochemical blood analysis, and evaluated the liver/body, spleen/body and intestine/body ratio. This evidence was known feeding with inactive Bacillus subtilis var. natto was able to improve the growth performance after 4 weeks, but not to affect the inflammatory biochemical blood parametres total protein (T-pro), albumin (Alb), Alb/T-pro ratio, creatine-phospho-kinase (CPK) and lactate dehydrogenase (LDH). Further, in the intestine microbiota, the Lactobacillaceae, Firmicutes, Chromatiales, and Rhodobacteria, was significantly higher than the control and the Firmicutes/Bacteroidetes ratio (F/B), which was indicated with a significantly increased. The intestine tissue metabolites OPLS-DA analysis indicated that the prominent bioactive metabolites changed. The peonidin-3-glucoside, l-Tyrosine, 1-Deoxy-1-(N6-lysino)-d-fructose was significantly increased. The feces metabolite OPLS-DA analysis indicated that the palmitelaidic acid, 5-KETE, tangeritin was significantly increased. In the transcriptome, the Gene Ontology (GO) analysis was found to enhance the intestine intestinal immune network. Combine of these evidence, feeding of the heat inactive Bacillus subtilis var. natto exactly improved the O. niloticus growth performance and regulation of the microbiota to promote the metabolites. In the transcriptome analysis, it was found to involve in the intestine immune phagosome response. Summarized of this study, the heat inactive Bacillus subtilis var. natto was reported to affect Nile tilapia intestine microbiota, and could positively regulate the intestine and fecal metabolites production to improve the intestine immune network.

Cell Differentiation Trajectory Predicts Prognosis and Immunotherapeutic Response in Clear Cell Renal Cell Carcinoma.

Clear cell renal cell carcinoma (ccRCC) is the main type of malignancy in kidney related to glucose metabolism. Primary single cell culture and single cell sequencing are novel research technologies. In this study, we explored the differentiation status of ccRCC cells and its significance in prognosis and immunotherapeutic response through bioinformatics. We characterized distinct differentiation states and differentiation-related genes (DRGs) in ccRCC cells through single cell RNA sequencing (scRNA-seq) analysis. Combined with bulk RNA-seq data, we classified patients into two clusters and found that this classification was closely correlated with patient prognosis and immunotherapeutic responses. Based on machine learning, we identified a prognostic risk model composed of 14 DRGs, including BTG2, CDKN1A, COL6A1, CPM, CYB5D2, FOSB, ID2, ISG15, PLCG2, SECISBP2, SOCS3, TES, ZBTB16, and ZNF704, to predict the survival rate of patients and then constructed a nomogram model integrating clinicopathological characteristics and risk score for clinical practice. In the study of immune checkpoints, we found that patients in the high-risk group had a disposition to get worse prognosis and better effects of immune checkpoint blocking therapies. Finally, we found the expression level of model DRGs was associated with a tumor-immune microenvironment (TIME) pattern and the response of 83 compounds or inhibitors was significantly different in the two risk groups. In a word, our study highlights the potential contribution of cell differentiation in prognosis judgment and immunotherapy response and offers promising therapeutic options for ccRCC patients.

Dietary of Lactobacillus paracasei and Bifidobacterium longum improve nonspecific immune responses, growth performance, and resistance against Vibrio parahaemolyticus in Penaeus vannamei.

This study investigated the effects of two probiotics, namely Lactobacillus paracasei and Bifidobacterium longum, as feed additives on growth performance, nonspecific immunity, immune-related gene expression, and disease resistance against Vibrio parahaemolyticus in Penaeus vannamei. The experimental diets were prepared using L. paracasei and B. longum at concentrations of 105 and 107 CFU/g; these diets were referred to as P5, P7, B5, and B7. After 8 weeks of the diets, regarding growth performance, the B7 group showed the highest weight gain rate (890.34 ± 103.65%), special growth rate (4.08 ± 0.19%), and feed conversion rate (1.52 ± 0.19%) compared with the other groups. Moreover, the total hemocyte counts were significantly increased (p < 0.05) in the P7 groups on day 14 during the 28-day feeding trial. The phagocytosis rate in all experimental groups was increased on day 14 and was persistently significantly activated to day 21, especially in the P7 and B5 group. The phagocytic index of the P7 group showed a significant increase on day 14 and persistent activation to day 21. In the analysis of respiratory burst activity and phenoloxidase activity, the P7 and B5 groups showed a significant increase on day 7 and persistent activation to day 21. The expression level of the immune-related genes of superoxide dismutase, clotting protein, Penaeidin2, Penaeidin3, Penaeidin4, anti-LPS factor, crustin, and lysozyme was significantly increased in the experimental groups, especially in the P7 group. Furthermore, the optimum conditions of feed additives were determined in challenge trials conducted using P7 and B5. Shrimps fed P7 and B5 showed an increased survival rate (72.73% and 66.67%) after the V. parahaemolyticus challenge. In sum, the results revealed that B. longum, as a feed additive at 107 CFU/g, enhanced growth performance. L. paracasei at 107 CFU/g and B. longum at 105 CFU/g can enhance nonspecific immune responses and immune-related gene expression, and 107 CFU/g L. paracasei has the highest resistance ability for V. parahaemolyticus. Thus, dietary supplementation with L. paracasei and B. longum may be a valuable approach in white shrimp aquaculture.

Let-7i-3p inhibits the cell cycle, proliferation, invasion, and migration of colorectal cancer cells via downregulating CCND1.

Dysregulated microRNAs are closely related to the malignant progression of colorectal cancer (CRC). Although abnormal let-7i-3p expression has been reported in various human cancers, its biological role and potential mechanism in CRC remain unclear. Therefore, the purpose of this study was to investigate the expression and regulation of let-7i-3p in CRC. Here, we demonstrated that let-7i-3p expression was significantly downregulated in three CRC cell lines while CyclinD1 (CCND1) was upregulated compared with the normal colon epithelial FHC cells. Moreover, bioinformatics and luciferase reporter assays revealed that CCND1 was a direct functional target of let-7i-3p. In addition, let-7i-3p overexpression or CCND1 silencing inhibited cell cycle, proliferation, invasion, and migration and diminished the activation of p-ERK in HCT116 cells. However, exogenously expressing CCND1 alleviated these effects. Taken together, our findings may provide new insight into the pathogenesis of CRC and let-7i-3p/CCND1 might function as new therapeutic targets for CRC.

Synergistic effects of dietary oxolinic acid combined with oxytetracycline on nonspecific immune responses and resistance against Vibrio parahaemolyticus infection of white shrimp (Penaeus vannamei).

This study investigated the synergistic effects of oxolinic acid (OA) combined with oxytetracycline (OTC) on white shrimp (Penaeus vannamei). Disk diffusion susceptibility testing was performed to analyze the sensitivity of Vibrio alginolyticus and Vibrio parahaemolyticus to different concentrations of OA and OTC. The results revealed that 50 mg OA/L combined with 50 mg OTC/L exhibited stronger antibacterial effects on V. alginolyticus and V. parahaemolyticus. The results of in vitro tests indicated that cotreatment with OA and OTC significantly reduced superoxide anion production and phenoloxidase activity, but not phagocytic activity. Subsequently, feeding trials were performed to investigate the immunomodulatory effects and bioaccumulation of dietary OA combined with OTC on shrimp. The healthy shrimp (15.13 ± 1.02 g) were divided into four groups: control, 100 mg OA/kg combined with 50 mg OTC/kg, 50 mg OA/kg combined with 100 mg OTC/kg, and 50 mg OA/kg combined with 50 mg OTC/kg. The shrimp were sampled to determine innate immunity parameters and residual OA and OTC levels in the muscle during a 28-day feeding regimen; the shrimp were fed the experimental diet from day 1 to day 5 and a commercial diet from day 6 to day 28. Residual OA levels were considerably higher in the group fed 100 mg OA/kg combined with 50 mg OTC/kg compared with the other groups and peaked on day 4. The residual OA levels of all the groups were below the detection limit after without providing OA. The residual OTC levels of the group fed 50 mg OA/kg combined with 100 mg OTC/kg were considerably higher from day 1 to day 4. The residual OTC levels in all the groups decreased rapidly and could not be detected on day 28. The administration of 50 mg OA/kg combined with 100 mg OTC/kg exerted the least effect on the white shrimp. Moreover, the survival rates of the treatment groups after the V. parahaemolyticus challenge were higher than those of the control group, especially the group fed 50 mg OA/kg combined with 100 mg OTC/kg. This result indicated that the synergistic effects of dietary OA and OTC are safe and effective. Combination therapy is a new method of antibiotic use in aquaculture.

Effects of dietary Lactobacillus reuteri and Pediococcus acidilactici on the cultured water qualities, the growth and non-specific immune responses of Penaeus vannamei.

This study discussed the effects of two types of lactic acid bacteria, Lactobacillus reuteri (L. reuteri) and Pediococcus acidilactici (P. acidilactici), on the growth and nonspecific immunity of Penaeus vannamei (P. vannamei) and developed probiotic applications for shrimp cultivation. This study incorporated two types of lactic acid bacteria in shrimp feed through spraying. The shrimps were grouped according to the type and concentration of effective bacteria incorporated into their feed. This research was separated into 3 individual feeding treatment as control, L. reuteri (Lr groups) and P. acidilactici (Pa groups). The shrimp was feeding with 103, 105, and 107 cfu/feed (g) L. reuteri namely as Lr3, Lr5, and Lr7. The shrimp was feeding with 103, 105, and 107 cfu/feed (g) P. acidilactici were named Pa3, Pa5, and Pa7, respectively. Through 8 weeks of feeding, the results revealed that the use of shrimp feed incorporated with lactic acid bacteria did not cause negative effects on water quality. The testing items include ammonia-nitrogen concentration, nitrite-nitrogen concentration, and total vibrio count in the water. In addition, the lactic acid bacteria concentration in the water were in the range of 1.33 ± 0.58 × 101 to 9.77 ± 1.34 × 102 cfu/mL. Observations of the growth performance of the white shrimps after 8 weeks of feeding revealed that both bacteria were beneficial to shrimp growth. In particular, group Lr7 had the highest percentage weight gain (107.99 ± 3.92%), special growth rate (1.93 ± 0.07%), feed conversion ratio (3.34 ± 0.05), and survival rate (97.22 ± 4.81%). Furthermore, observations of the nonspecific immunity reactions of the white shrimps after 4 weeks of feeding indicated that on day 4, the total number of haemocyte in shrimps in groups Lr5, Lr7, Pa3, and Pa5 significantly increased. On days 1 and 4, the phenoloxidase activity and superoxide axion production rates of the Lr group and Ls group increased. This phenomenon was the most significant in group Lr7, and the effect continued until day 28. After day 7, the phagocytic rate of groups Lr5 and Lr7 significantly increased. In addition, Lr and Pa groups exhibited significant increases in the phagocytic index after days 4 and 14, respectively. This phenomenon was also the most significant in group Lr7.

Transdermal Composite Microneedle Composed of Mesoporous Iron Oxide Nanoraspberry and PVA for Androgenetic Alopecia Treatment.

The transdermal delivery of therapeutic agents amplifying a local concentration of active molecules have received considerable attention in wide biomedical applications, especially in vaccine development and medical beauty. Unlike oral or subcutaneous injections, this approach can not only avoid the loss of efficacy of oral drugs due to the liver's first-pass effect but also reduce the risk of infection by subcutaneous injection. In this study, a magneto-responsive transdermal composite microneedle (MNs) with a mesoporous iron oxide nanoraspberry (MIO), that can improve the drug delivery efficiency, was fabricated by using a 3D printing-molding method. With loading of Minoxidil (Mx, a medication commonly used to slow the progression of hair loss and speed the process of hair regrowth), MNs can break the barrier of the stratum corneum through the puncture ability, and control the delivery dose for treating androgenetic alopecia (AGA). By 3D printing process, the sizes and morphologies of MNs is able to be, easily, architected. The MIOs were embedded into the tip of MNs which can deliver Mx as well as generate mild heating for hair growth, which is potentially attributed by the expansion of hair follicle and drug penetration. Compared to the mice without any treatments, the hair density of mice exhibited an 800% improvement after being treated by MNs with MF at 10-days post-treatment.

Effects of medicinal herbs "Plantago asiatica", "Houttuynia cordata" and "Mentha haplocalyx" on non-specific immune responses of cobia (Rachycentron canadum).

This study investigated the effects of orally administered Plantago asiatica, Houttuynia cordata, and Mentha haplocalyx on the growth and nonspecific immune responses of cobia (Rachycentron canadum). The nonspecific immune parameters assessed were weight gain, feed conversion ratio, superoxide anion (O2-) production, superoxide dismutase (SOD) activity, phagocytic rate, phagocytic index, lysozyme activity, serum albumin and globulin, and albumin:globulin (A/G) ratio. The growth experiment indicated that 6-week dietary treatments did not significantly affect on the growth of cobia. Nonspecific immune responses showed that O2- production, SOD and lysozyme activity, and phagocytosis were significantly increased after the oral administration of P. asiatica and H. cordata, and the serum albumin:globulin ratio (A/G) gradually decreased. In this study, treatment of the Mentha haplocalyx on the cobia didn't present with the inducing of the phagocytosis ability compared with the treatment of P. asiatica and H. cordata. We suggest that oral administration of the 10 g/kg or 20 g/kg of the P. asiatica and H. cordata is exactly inducing the phagocytosis, ROS production, lysozyme activity and SOD production in the cobia.

Could structural and noncompensatory Lenke 3 and 4C lumbar curves be nonstructural and compensatory? Lenke 1, 2, 3, and 4 curve types were similar and could be considered collectively as a single indication for selective thoracic fusion.

STUDY DESIGN: Retrospective radiographical review. OBJECTIVE: To demonstrate that the structural and noncompensatory Lenke 3 and 4C lumbar curves could be nonstructural and compensatory. SUMMARY OF BACKGROUND DATA: Historically, Lenke 3 and 4C curves were not recommended for selective thoracic fusion (STF) because the lumbar curve was considered structural and noncompensatory. However, consecutive series of Lenke 3 and 4C curves suggest successful treatment with STF. METHODS: Between 2001 and 2004, 2005 and 2008, and 2010 and 2012, 3 consecutive series of 108, 134, and 78 surgically treated Lenke 1, 2, 3, and 4C curves were reviewed, respectively. The coronal curve criteria for the curves treated with STF during each period were lumbar side bending Cobb angle less than 25° and meeting the Lenke ratio criteria, lumbar side bending Cobb angle 35° or less, and lumbar side bending Cobb angle 45° or less, respectively. The sagittal curve criteria for STF during each period was absence of junctional thoracolumbar kyphosis 20° or more between T10 and L2. The technique used for STF was the Guan-Din method. Radiographs of all the curves treated with STF were analyzed before and after surgery. RESULTS: Optimal instrumented thoracic and compensatory lumbar correction was obtained for all Lenke 1, 2, 3, and 4C curves treated with STF in each period. As the coronal criteria for STF were broadened, the extent of feasibility of STF was expanded and the rate of STF increased. Although Cobb angle, apical vertebral translation, and apical vertebral rotation magnitudes of Lenke 3 and 4C curves were larger and more severe than those of Lenke 1 and 2C curves, optimal compensatory correction could still be obtained for Lenke 3 and 4C curves. CONCLUSION: The structural and noncompensatory Lenke 3 and 4C lumbar curves were proven to be nonstructural and compensatory. Lenke 1, 2, 3, and 4C curves have similar natures and similar responses to the same technique (Guan-Din method) used for STF and could be considered collectively as a single indication for STF. The extent of feasibility of STF could be expanded from Lenke 1 and 2 curves to Lenke 1, 2, 3, and 4 curves. LEVEL OF EVIDENCE: 2.

Guan-Din method: a novel surgical technique for selective thoracic fusion to maximize the rate of selective thoracic fusion and compensatory correction.

STUDY DESIGN: Retrospective radiographical review. OBJECTIVE: To evaluate the outcome of selective thoracic fusion (STF) by using the Guan-Din method for the treatment of major thoracic compensatory lumbar (MTCL) curves. SUMMARY OF BACKGROUND DATA: Performing STF for MTCL curves is to minimize the loss of lumbar motion and the risk of lumbar degeneration or pain. Surgical treatment of MTCL curves aims to maximize the rate of STF for MTCL curves while optimizing instrumental thoracic and compensatory lumbar correction. The Guan-Din method has been demonstrated to be able to enhance the lumbar curve's capacity for spontaneous correction and broaden the current curve criteria of MTCL curves for STF. METHODS: Between 2004 and 2010, 510 consecutive surgically treated MTCL curves were reviewed. Of these MTCL curves, who met the criteria of lumbar side bending Cobb 35° or less and without global thoracic hyperkyphosis and/or thoracolumbar kyphosis (T10-L2 ≤20°), were treated with STF using the Guan-Din method. Radiographs were analyzed before surgery, immediately after surgery, and at the most recent follow-up (range, 2-8 yr). RESULTS: Curve types of 510 MTCL curves according to Lenke system were as follows: 1A (n = 91), 2A (n = 74), 3A (n = 6), 4A (n = 2), 1B (n = 93), 2B (n = 34), 3B (n = 8), 4B (n = 5), 1C (n = 84), 2C (n = 26), 3C (n = 72), and 4C (n = 15). Of the 510 MTCL curves, 458 (90%) curves were treated with STF. A mean 73% thoracic correction and 63% lumbar correction was obtained at the most recent follow-up. Of the 197 surgically treated MTCL curves with a lumbar C modifier, 148 (75%) curves that contained 57 Lenke 1C and 2C curves and 40 Lenke 3C and 4C curves that did not meet Lenke curve criteria for STF, were successfully treated with STF. A mean 67% thoracic correction and 57% lumbar correction was obtained at the most recent follow-up. The rate of STF and the magnitude of correction of MTCL curves in this study were significantly greater than those in all other reports. No significant change in global coronal and sagittal imbalance was observed. CONCLUSION: The rate of STF and the compensatory correction of MTCL curves could be maximized by using the Guan-Din method as the method for STF. LEVEL OF EVIDENCE: 4.

Transmembrane protein 166 regulates autophagic and apoptotic activities following focal cerebral ischemic injury in rats.

Transmembrane protein 166 (TMEM166) is a lysosomal/endoplasmic reticulum-associated protein found in various species where it acts as a regulator of programmed cell death, mediating both autophagy and apoptosis. In the present study, we investigated the role of TMEM166 following MCAO injury in rats to determine whether the structural damages following injury were orchestrated in part by TMEM166. One hundred and fifty six male Sprague-Dawley rats were randomly divided into 4 groups: Sham, MCAO, MCAO+control siRNA, MCAO+TMEM166 siRNA. Outcomes were measured including mortality rate, brain edema, BBB disruption, and neurobehavioral testing. Western blotting techniques measured the expression of key pro-autophagic and apoptotic proteins such as TMEM166, Beclin-1, cleaved casepase-3 and Bcl-2/Bax. The study found that TMEM166 siRNA treatment significantly reduced the mortality rate, cerebral edema, neurobehavioral deficits, and BBB disruption as measured by Evan's blue assay following MCAO injury. Immunohistochemical staining and western blotting analysis demonstrated an increased expressions of TMEM166, Beclin-1, LC3, cleaved casepase-3 and Bcl-2/Bax in the infarcted areas. This study suggests that TMEM166 induces autophagy and apoptosis may in fact play a significant role in cell death following MCAO injury and its mediation may be through the crosstalk of Bcl-2. By blocking the activity of TMEM166 using siRNA, we were able to prevent the cell loss that occured following cerebral ischemia injury. This translated into a preservation of functional integrity and an improvement in mortality.

Tai Chi pedicle screw placement for severe scoliosis.

STUDY DESIGN: Retrospective. OBJECTIVE: To evaluate the clinical safety and accuracy of the Tai Chi ((Equation is included in full-text article.)) technique for placing pedicle screws, without intraoperative radiographic imaging, in severe scoliotic spines. SUMMARY OF BACKGROUND DATA: The current techniques for pedicle screw placement have a number of drawbacks in cases of severe scoliosis, including difficulty or impossibility to use, delayed operative time, requiring the presence of trained personnel for the duration of the surgery, high cost issues, increased radiation exposure, and technical challenges. No previous report has described the application of the Tai Chi pedicle screw placement technique for severe scoliosis. MATERIAL AND METHODS: Between 2006 and 2008, the cases of 39 consecutive patients with severe scoliosis (Cobb angle >100 degrees) who underwent posterior correction and stabilization (from T1 to L5) using 992 transpedicular screws were examined. The mean patient age was 25.7 (range, 11 to 63) years at the time of surgery. Pedicle screws were inserted by the Tai Chi technique using anatomic landmarks and preoperative radiographs as a guide. Tai Chi drilling fully utilizes the natural anatomic and physical characteristics of pedicles and unconstrained circular force. By nature, a drill bit driven by unconstrained circular force would migrate within the pedicle along a path of least resistance, advancing along the central cancellous bone tunnel spontaneously. Accurate drilling was achieved by following the nature and sticking to the hand sensation when the drill bit broke through the cancellous bone. The total time for inserting all pedicle screws in each case was recorded. Postoperative computed tomography scans were performed to evaluate the position of the inserted pedicle screws. The screw position was classified as "in" or "out." The distance of perforation was measured. RESULT: The average Cobb angle was 127 degrees (range, 100 to 153 degrees). The number of screws inserted at each level were as follows: T1 (n=10), T2 (n=34), T3 (n=46), T4 (n=53), T5 (n=61), T6 (n=69), T7 (n=75), T8 (n=76), T9 (n=76), T10 (n=77), T11 (n=76), T12 (n=78), L1 (n=77), L2 (n=68), L3 (n=56), L4 (n=38), and L5 (n=22). There were 923 (93%) "in" screws and 69 (7%) "out" screws. The overall accuracy of screw placement was 93%. There were no neurological, vascular, or visceral complications. No screws required postoperative repositioning. The average time for pedicle screw placement was 73 seconds. CONCLUSIONS: Our findings suggest that the Tai Chi pedicle screw placement technique, which does not require intraoperative radiographic imaging, is an accurate, reliable, safe, and time-saving method of placing pedicle screws in severe scoliotic spines.

Broader curve criteria for selective thoracic fusion.

STUDY DESIGN: Retrospective radiographic review. OBJECTIVE: To evaluate the outcome of selective thoracic fusion (STF) by using cantilever bending technique (CBT) and the direct vertebral rotation (DVR) technique for major thoracic-compensatory lumbar (MTCL) curves selected by new curve criteria, which are broader than Lenke curve criteria for STF. SUMMARY OF BACKGROUND DATA: Surgical treatment of MTCL curves aims to maximize the number of MTCL curves that can be treated with STF and optimize instrumented thoracic and spontaneous lumbar correction. Comparing current guidelines for STF shows that the surgical technique utilized for STF may affect the curve criteria for MTCL curves for successful STF and thoracic and lumbar correction. METHODS: Seventy-eight consecutive idiopathic scoliosis patients with major thoracic-compensatory "C" modifier lumbar curves who met the following three criteria: (1) main thoracic curve (MT) to compensatory lumbar curve (CL) ratios of Cobb magnitude and apical vertebral translation (AVT) greater than one; (2) MT/CL ratio of flexibility less than one; (3) Cobb magnitude of lumbar curve less than 35° on side bending, were treated with STF by using CBT and DVR. Radiographs were analyzed before surgery, immediately after surgery, and at the most recent follow-up (range, 2-5 years). RESULTS: All 78 MTCL curves were successfully treated with STF by using CBT and DVR. A mean 61% thoracic correction was matched by 55% lumbar correction at the most recent follow-up. Spontaneous correction of lumbar AVT occurred in all patients. Global coronal imbalance was common before surgery (mean, 14 mm) and remained so after surgery (mean, 12 mm). There were 49 MTCL curves that did not meet Lenke curve criteria for STF. All were successfully treated with STF by using CBT and DVR. Among these 49 MTCL curves, there were 14 Lenke 1C and 18 Lenke 2C curves with one or two, or all of MT/CL ratios of Cobb magnitude, AVT, and apical vertebral rotation of 1.2 or less, and 6 Lenke 3C and 11 Lenke 4C curves with the Cobb magnitude of residual lumbar curve on side bending between 25° and 35°. CONCLUSION: CBT and DVR can broaden the current curve criteria of MTCL curves for STF to have more MTCL curves treatable with STF and optimize instrumented thoracic and spontaneous lumbar correction. A more effective surgical technique can not only improve instrumented thoracic and spontaneous lumbar correction but also can broaden the MTCL curve criteria for STF to have more MTCL curves treatable with STF.

Quality control of reconstructed sagittal balance for sagittal imbalance.

STUDY DESIGN: Prospective radiographic study. OBJECTIVE: To investigate the feasibility of controlling quality of reconstructed sagittal balance for sagittal imbalance. SUMMARY OF BACKGROUND DATA: Patients with sagittal imbalance cannot walk or stand erect without overwork of musculature because of compromised biomechanical advantage. The result is muscle fatigue and activity-related pain. During reconstructive surgery, restoration of optimal sagittal balance is crucial for obtaining satisfactory clinical results. However, there is no way to control quality of reconstructed sagittal balance before or during surgery. METHODS: A method was developed to determine the lumbosacral curve in a way that theoretically would bring sagittal balance to an ideal state by calculation and simulation for each patient before surgery and then template rods of the curve and a blueprint were made accordingly for operative procedures. Ninety-four consecutive patients with sagittal imbalance due to lumbar kyphosis were treated for intractable pain and then followed up for a mean of 4.3 years. Radiographs were analyzed before surgery, 2 months after surgery, and at most recent follow-up. RESULTS: The mean estimated values of L1-S1 lordosis, sacral inclination angle, sacrofemoral distance, and distribution of L1-S1 lordosis at the closing-opening wedge osteotomy site and L4-S1 segments were 30.8°, 24.6°, 0 mm, 16.1% (-5°), and 62% (-19°), respectively. The mean reconstructed values were 41.1°, 23.3°, 3.9 mm, 41% (-17°), and 46% (-19°), respectively. There were significant differences between estimated and reconstructed values of L1-S1 lordosis and the percentage of distributions; however, there was no significant difference between the estimated and reconstructed magnitude of L4-S1 lordosis, sacral inclination angle, and sacrofemoral distance. A properly oriented pelvis can be brought nearly directly above the hip axis. The mean sagittal global balance, represented by the distance between the vertical line through the hip axis and sacral promontory, improved from 61.4 mm before surgery to 3.9 mm 2 months after surgery, and 1.3 mm at final follow-up. Normal sagittal global balance was reconstructed and maintained. The mean sagittal spinal balance measured as the horizontal distance between the C7 sagittal plumb line and the posterior superior corner of S1 improved from 97.4 mm before surgery to 11 mm 2 months after surgery. However, there was significant loss of sagittal spinal balance to 25.4 mm at the fi nal visit. Normal sagittal spinal balance was reconstructed and appeared to be maintained. The magnitude of T1-T12 kyphosis compensated from 13° before surgery to 25.2° 2 months after surgery and 34.5° at fi nal follow-up. CONCLUSIONS: Quality control of the reconstructed sagittal balance for sagittal imbalance is possible. Correctly orienting the pelvis, reconstructed by the restoration of enough L1-S1 lordosis with adequate distribution at L4-S1 segments, is a matter of critical importance for optimizing reconstructed sagittal balance. The correctly oriented pelvis can be determined before surgery. Preventing junctional fracture and persistent rehabilitation of surgically injured lumbar extensor musculature are crucial for maintaining the reconstructed sagittal balance.