Establishment of the Lunar Phase Morphological Classification for Cervical Spinal Canal.

STUDY DESIGN: Retrospective clinical trial. PURPOSE: To establish a morphological classification of the cervical spinal canal using its parameters. OVERVIEW OF LITERATURE: Cervical spine computed tomography (CT) data of 200 healthy volunteers in 2 years were analyzed. The morphology of the spinal cord was also analyzed. METHODS: The median sagittal diameter and transverse diameter of the spinal canal from C2 to C7 were measured on CT images. The ratio of the median sagittal diameter to the transverse diameter was calculated. Accordingly, the spinal canal shape of each segment was classified into four, and the specific criteria of lunar phase classification were determined through linear discriminant analysis based on the ratio of the median sagittal diameter to the transverse diameter. The inter-rater reliability of the classification was explored using Kappa coefficients. Finally, the morphology of the different segments of the cervical spinal canal in healthy volunteers was revised and compared. RESULTS: According to the ratio of the median sagittal diameter and the transverse diameter of the cervical spinal canal, the lunar phase classification of the cervical bony spinal canal was determined as follows: full-moon >0.65, 0.55< convex-moon ≤0.65, 0.46≤ quarter-moon ≤0.55, and residual-moon <0.46. The Kappa values of C2-C7 were 0.851, 0.958, 0.823, 0.927, 0.793, and 0.946, and the Kappa value of all C2-C7 segments was 0.854 that mainly presented two forms of full-moon (76.5%) and convex-moon (23.0%). A quarter-moon spinal canal was mainly distributed in C3, C4, C5, and C6; a residual-moon spinal canal was mainly distributed in C4 and C5; and the morphological distribution of C4 and C5 were similar (p>0.05). The frequency of the spinal canal of the residual-moon type was the highest, and the full-moon (6.5%) and residual-moon (7.5%) types of C7 were rare. CONCLUSIONS: The morphological classification of the cervical spinal canal was established to present anatomical variations. The classification showed good inter-rater reliability.

An advanced treatment process for 3-high wastewater discharged from crude oil storage tanks.

The wastewater discharged from crude oil storage tanks (WCOST) contains high concentrations of salt and metal iron ions, and high chemical oxygen demand (COD). It belongs to "3-high" wastewater, which is difficult for purification. In this study, WCOST treatments were comparatively investigated via an advanced pretreatment and the traditional coagulation-microfiltration (CMF) processes. After WCOST was purified through the conventional CMF process, fouling occurred in the microfiltration (MF) membrane, which is rather harmful to the following reverse osmosis (RO) membrane unit, and the effluent featured high COD and UV254 values. The analysis confirmed that the MF fouling was due to the oxidation of ferrous ions, and the high COD and UV254 values were mainly attributable to the organic compounds with small molecular sizes, including aromatic-like and fulvic-like compounds. After the pretreatment of the advanced process consisting of aeration, manganese sand filtration, and activated carbon adsorption in combination with CMF process, the removal efficiencies of organic matter and total iron ions reached 97.3% and 99.8%, respectively. All the water indexes of the effluent, after treatment by the advanced multi-unit process, meet well the corresponding standard. The advanced pretreatment process reported herein displayed a great potential for alleviating the MF membrane fouling and enhanced the lifetime of the RO membrane system in the 3-high WCOST treatment.

Identification and functional analysis of a chemosensory protein from Bactrocera minax (Diptera: Tephritidae).

BACKGROUND: Olfaction has an indispensable role in insect behavior, enabling location of suitable host plants and oviposition sites, finding mates and evasion of natural enemies. Chemosensory proteins (CSPs) function to screen external odorants and transport them to olfactory receptor neurons, thereby increasing the sensitivity of the olfactory system. At present, CSP genes have been identified in many insect species, but there are relatively few studies on the function of CSP, especially in Tephritidae. RESULTS: In this study, we sequenced and analyzed 12 transcriptomes of Bactrocera minax and identified five CSP genes. The results of polymerase chain reactions with reverse transcription showed that BminCSP3 was highly expressed only in antennae. Results from competitive binding experiments showed that BminCSP3 has good binding ability to citral compared with 23 other volatile organic compounds. The docking model with citral showed hydrogen bond formation with residues (ARG97); however, no hydrogen bonds were formed in the docking of five other ligands (furfuryl alcohol, linalool, cis-3-hexenyl acetate, (R)-(+)-limonene and (+)-carvone). Electroantennogram (EAG) analyses revealed that citral was active in B. minax at the antennal level, and the EAG response value of female adults was significantly higher than that of male adults. Furthermore, the results of behavioral bioassays showed that females were significantly attracted to citral. CONCLUSION: Our results suggest that BminCSP3 plays an important role in the recognition of citral by B. minax adults. © 2022 Society of Chemical Industry.

Interobserver and Intraobserver Reproducibility and Reliability of the Huashan Clinical Classification System for Hirayama Disease.

Purpose: The Huashan clinical classification system for Hirayama disease has recently been proposed and has been found useful for diagnosis and treatment. So far, however, there has been little in-depth evaluation of its reliability. Thus, this study aimed to assess the reproducibility and reliability of the system. Methods: Patients diagnosed with Hirayama disease between 2019 and 2020 were recruited. Seven spine surgeons from four different institutions, including an experienced group of three and an inexperienced group of four, were trained as observers of the Huashan clinical classification system for Hirayama disease, and these surgeons classified the recruited patients using the system. Then, 2 months later, they repeated the classification on the same patients in a different order. The interobserver and intraobserver agreement between the results was analyzed using percentage agreement and weighted kappa (κ) statistics. Results: A total of 60 patients were included in the analysis. For all the observers, experienced observers, and inexperienced observers, the agreement percentages were, respectively, 78.5% (κ = 0.76), 80.0% (κ = 0.78), and 78.9% (κ = 0.77), indicating substantial interobserver reproducibility. For distinguishing typical (Types I and II) and atypical (Type III) Hirayama disease among the different groups of observers, the percentage agreement ranged from 95.6 to 98.9% (κ = 0.74-0.92), indicating substantial to nearly perfect reproducibility. For suggesting conservative treatment (Types I and III) or surgery (Type II), the percentage agreement ranged from 93.3 to 96.4% (κ = 0.81-0.90), indicating nearly perfect reproducibility. As for intraobserver agreement, the percentage agreement ranged from 68.3 to 81.7% (κ = 0.65-0.79), indicating substantial reliability. Conclusion: The Huashan clinical classification system for Hirayama disease was easy to learn and apply in a clinical environment, showing excellent reproducibility and reliability. Therefore, it would be promising to apply and promote this system for the precise and individualized future treatment of Hirayama disease.

Characterization of a specific odorant receptor for linalool in the Chinese citrus fly Bactrocera minax (Diptera: Tephritidae).

Insect sensing of odorants plays important roles in various behaviors, including host location, mate attraction, and oviposition site selection. The odorant receptor (OR) is a key protein in insect environmental odor recognition. Most Diptera studies of ORs have focused on Drosophila and mosquitos, so there little known about ORs in the agricultural pest insects Tephritidae. To understand the olfactory recognition mechanism of Bactrocera minax, we sequenced and analyzed 12 B. minax transcriptomes to identify a total of 59 OR genes. Semi-quantitative reverse transcription PCR (RT-PCR) showed that several BminORs were highly expressed in antennae. Available with a complete open reading frame and expressed in the antennae of both sexes at a higher level than those of other BminORs, BminOR24 was selected for further functional analyses. BminOR24/BminOrco expressed in Xenopus oocytes responded significantly to linalool. The identification of B. minax OR genes lays a foundation for further functional studies of OR genes, and functional characterization of BminOR24 provides insight for improving methods for controlling B. minax, a devastating pest insects.

Cloning and Functional Characterization of Three Odorant Receptors From the Chinese Citrus fly Bactrocera minax (Diptera: Tephritidae).

Insect olfactory sensing is crucial for finding food, mating, and oviposition preference. Odorant receptors (ORs) play a central role in the transmission of odorant signals into the environment by the peripheral olfactory system. Therefore, the identification and functional study of ORs are essential to better understand olfactory mechanisms in insects. OR studies on Diptera insects are primarily performed on Drosophila and mosquitoes, but few studies have been reported in Tephritidae. In this study, we examined three candidate ORs (BminOR3, BminOR12, and BminOR16) from Bactrocera minax. Our analysis of tissue expression revealed that the three BminORs were expressed in the antennae, with no difference between the male and female. In in vitro heterologous expression system of Xenopus oocytes. BminOR3/BminOrco responded strongly to 1-octen-3-ol, BminOR12/BminOrco responded to eight compounds [methyl salicylate, benzaldehyde, (Z)-3-hexenyl acetate, butyl acrylate, butyl propionate, 1-octanol, (S)-(+)-carvone and benzyl alcohol], and BminOR16/BminOrco slightly responded to undecanol. Our results concluded that BminOR3, BimOR12, and BminOR16 could play an important role in host-finding and oviposition positioning in B. minax.

Analysis of the Effect of Surgical Treatment for the Patients with Hirayama Disease from the Perspective of Cervical Spine Sagittal Alignment.

OBJECTIVE: This study was carried out to analyze the surgical effect of cervical spine sagittal alignment for patients with Hirayama disease (HD). METHODS: Forty-four subjects were retrospectively analyzed for the parameters of cervical spine sagittal alignment. The case group consisted of 23 patients with HD, whereas the control group consisted of 21 healthy adolescent subjects. Pre- and postoperative cervical spine sagittal parameters of the patients with HD were collected; the cervical sagittal parameters of the healthy adolescent subjects were also collected. Sagittal alignment parameters were compared between the patients with HD and the healthy adolescent subjects, and between the pre- and postoperative parameters for the patients with HD. RESULTS: Forty-four subjects completed the follow-up, with the average follow-up period being 18.0 months. No significant differences were detected between the HD and control groups for clinical parameters (P > 0.05). The preoperative HD group had smaller values compared with the control group in the sagittal parameters of C2-7 cervical lordosis (CL) angle, T1 slope, thoracic inlet angle (TIA), and cervical tilt angle (P < 0.05). For the patients with HD, the preoperative values were smaller compared with the postoperative HD values for the parameters of C2-7 CL angle, T1 slope, and cervical tilt angle (P < 0.05). We found no significant differences between the postoperative patients with HD and the healthy subjects, including C2-7 CL angle, C2-7 sagittal vertical axis, T1 slope, TIA, neck tilt angle, cervical tilt angle, and cranial tilt angle (P > 0.05). CONCLUSIONS: Patients with HD have sagittal imbalance of the cervical spine compared with age-matched healthy adolescent subjects, and surgical treatment could correct the sagittal imbalance.

The evaluation on neural status of cervical spinal cord in normal and Hirayama disease using diffusion tensor imaging.

PURPOSE: To explore the changes in diffusion tensor imaging (DTI) parameters in cervical spinal cord in Hirayama disease (HD) patients and healthy volunteers and to compare these parameters between cervical flexion and neutral positions in HD patients. METHODS: Seventeen male patients with HD and eleven healthy young males were included to receive DTI scans in cervical flexion and neutral positions. The FA and ADC values of different levels were measured based on the region of interest drawn on the mid-sagittal plane. The dynamic compressed level's parameters were defined as the lowest and the second lowest FA and the highest and the second highest ADC, respectively. The clinical assessment of patients was obtained using their disabilities of the arm, shoulder and hand (DASH) scores. RESULTS: For the HD patients, the FA values in the cervical flexion position were lower and the ADC values were much higher than those in the cervical neutral position. Compared with the controls, the ADC values were significantly higher in the lower levels (C5/6-C7/T1) and the FA values obviously lower at C7/T1 in HD patients in cervical neutral position. The FA and ADC values of the dynamic compressed level in HD patients deviated significantly from the average of the lower levels in controls. Both the FA and ADC values of the dynamic compressed level correlated with the DASH scores (FA, R2 = 0.520, P = 0.001; ADC, R2 = 0.421, P = 0.005). CONCLUSIONS: DTI parameters can support a hypothesis of dynamic cervical flexion compression and noninvasively reveal the neural status of HD patients. These slides can be retrieved under Electronic Supplementary Material.

Enhanced Enzymatic Hydrolysis and Structural Features of Corn Stover by NaOH and Ozone Combined Pretreatment.

A two-step pretreatment using NaOH and ozone was performed to improve the enzymatic hydrolysis, compositions and structural characteristics of corn stover. Comparison between the unpretreated and pretreated corn stover was also made to illustrate the mechanism of the combined pretreatment. A pretreatment with 2% (w/w) NaOH at 80 °C for 2 h followed by ozone treatment for 25 min with an initial pH 9 was found to be the optimal procedure and the maximum efficiency (91.73%) of cellulose enzymatic hydrolysis was achieved. Furthermore, microscopic observation of changes in the surface structure of the samples showed that holes were formed and lignin and hemicellulose were partially dissolved and removed. X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) and Cross-Polarization Magic Angle Spinning Carbon-13 Nuclear Magnetic Resonance (CP/MAS 13C-NMR) were also used to characterize the chemical structural changes after the combined pretreatment. The results were as follows: part of the cellulose I structure was destroyed and then reformed into cellulose III, the cellulose crystal indices were also changed; a wider space between the crystal layer was observed; disruption of hydrogen bonds in cellulose and disruption of ester bonds in hemicellulose; cleavage of bonds linkage in lignin-carbohydrate complexes; removal of methoxy in lignin and hemicellulose. As a result, all these changes effectively reduced recalcitrance of corn stover and promoted subsequent enzymatic hydrolysis of cellulose.

Sodium selenite (Na2SeO3) induces apoptosis through the mitochondrial pathway in CNE-2 nasopharyngeal carcinoma cells.

This study investigated the effect of sodium selenite (Na2SeO3) on proliferation, cell cycle, apoptosis as well as the underlying mechanism in CNE-2 nasopharyngeal carcinoma (NPC) cells. The CNE-2 cell line was treated with different concentrations of Na2SeO3, and the effects of Na2SeO3 on cell viability and proliferation were evaluated using Cell Counting kit-8 (CCK-8) assay. Cellular apoptosis and cell cycle were evaluated by flow cytometry following Annexin V­FITC/PI double staining and PI single staining respectively; nuclei morphology stained with DAPI and Hoechst 333258 was observed under a fluorescence microscope, while DNA fragmentation was detected by agarose gel electrophoresis. The mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were analyzed using fluorescent staining assays. Expression of Bcl-XL, Bax, Bak, and caspase-3 activation were examined by western blotting. The results showed that Na2SeO3 inhibited proliferation and induced apoptosis of CNE-2 cells in a dose- and time-dependent manner. Na2SeO3 at low concentrations induced cell cycle arrest at S phase, while high concentrations of Na2SeO3 induced cell cycle arrest at G0/G1 phase. Furthermore, Na2SeO3 increased ROS level and decreased MMP, upregulated caspase-3 activity and the expression of Bak and Bax but simultaneously downregulated Bcl-XL. In conclusion, our studies demonstrated that Na2SeO3 had significant anti-proliferative and apoptosis-inducing effects via arresting cell cycle and regulating mitochondria-mediated intrinsic caspase pathway in CNE-2 NPC cells, suggesting that Na2SeO3 might have therapeutic potentials in the treatment of NPC.

EGCG decreases the expression of HIF-1α and VEGF and cell growth in MCF-7 breast cancer cells.

PURPOSE: To investigate the effects of epigallocatechin-3-gallate (EGCG) on the expression of HIF-1α and vascular endothelial growth factor (VEGF) and cell growth in MCF-7 breast cancer cells. METHODS: MCF-7 human breast cancer cells were pretreated with different concentrations of EGCG (25, 50, 100 mg/L) for 48 h. The growth and proliferation of cells were analyzed by trypan blue staining in the pretreated MCF-7 cells. Furthermore, mRNA expression of HIF-1α and VEGF was detected by reverse transcriptase polymerase chain reaction (RT-PCR) analysis in the pretreated MCF-7 cells. Protein expression of HIF-1α was detected by Western blot, and the secreted protein level of VEGF in the supernatant of the culture medium was analyzed by enzyme linked immuno- sorbent assay (ELISA) in the MCF-7 cells pretreated with different concentrations of EGCG. RESULTS: Cell growth decreased dramatically in MCF-7 cells treated with different concentrations of EGCG, compared with untreated (control) cells. Moreover, protein expression of HIF-1α and VEGF declined in a dose-dependent manner in MCF-7 cells pretreated with increasing concentrations of EGCG. CONCLUSIONS: EGCG inhibits cell growth and proliferation of MCF-7 breast cancer cells, possibly by inhibiting the protein expression of HIF-1α and VEGF.

Construction and identification of the recombinant lentiviral expression vector targeting human BAX inhibitor-1 gene.

PURPOSE: The aim of this study was to construct a recombinant lentiviral expression vector targeting human BAX inhibitor- 1(BI-1) gene and observe its expression in NIH3T3 cells. METHODS: Human BI-1 gene was amplified by polymerase chain reaction (PCR), and then cloned into the vector pLCMV- IG using DNA recombinant technique. After the inserted sequences in the recombinant plasmids were identified by PCR, and double digesting and DNA sequencing analysis, the recombinant lentivirus was packaged and administered into NIH3T3 cells. The BI-1 mRNA and protein expression were examined by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. RESULTS: PCR double digesting analysis and DNA sequencing confirmed that the BI-1 DNA sequences were successfully inserted into the lentiviral vectors. After transfection with the recombinant lentivirus, BI-1 expression in NIH3T3 cells was significantly increased at both mRNA and protein levels. CONCLUSION: The lentiviral vector expressing BI-1 has been successfully constructed, which allowed for the subsequent analysis of the role of BI-1 in cell growth and transduction.

Ghrelin attenuates intestinal ischemia/reperfusion injury in mice by activating the mTOR signaling pathway.

Intestinal ischemia/reperfusion (I/R) injury is a serious condition in intensive care patients, resulting in severe inflammation and remote organ damage. The activation of the mammalian target of rapamycin (mTOR)/p70 ribosomal S6 kinase (p70S6K) signaling pathway exerts protective effect against ischemia/reperfusion injury. Ghrelin, an orexigenic hormone, inhibits the release of pro-inflammatory cytokines, such as interleukin (IL)-1ß, tumor necrosis factor-α and IL-6. In this study, we investigated the effects of ghrelin on gut I/R injury and the regulation of the mTOR/p70S6K signaling pathway following gut I/R injury in mice. C57BL/6 mice underwent superior mesenteric artery occlusion for 45 min, followed by reperfusion for 4 h. Ghrelin was administered at the onset of reperfusion. We assessed survival, organ injury variables, pro-inflammatory cytokine expression and observed the histological changes of the small intestine and lungs. Our results revealed that the administration of ghrelin inhibited the release of certain pro-inflammatory cytokines, reduced neutrophil infiltration, attenuated organ injury and improved survival following gut I/R injury. The administration of D-Lys-GHRP6, a specific ghrelin receptor antagonist, to a certain extent, counteracted the protective effects of ghrelin in gut I/R-induced organ injury and mortality. To determine whether the beneficial effects of ghrelin following gut I/R injury are associated with the mTOR/p70S6K signaling pathway, the phosphorylation levels of mTOR and p70S6K were detected by western blot analysis. Our results revealed that mTOR and p70S6K phosphorylation increased in the tissue from the small intestine and pulmonary tissue in the animals treated with ghrelin. These findings suggest that ghrelin attenuates organ injury following gut I/R by promoting the activation of the mTOR/p70S6K signaling pathway.