Neuro-Interventional Diagnosis and Treatment Using the Right Transradial Approach in Patients with Aberrant Right Subclavian Artery.

OBJECTIVE: To assess the effectiveness and safety of neurological interventions using the right transradial approach (R-TRA) in patients with aberrant right subclavian artery (ARSA). METHODS: We retrospectively analyzed cases that underwent cerebral angiography and interventions at Huangpi District People's Hospital from January 2023 to July 2023. Out of 335 cases, 5 patients with ARSA were identified. RESULTS: All 5 cases underwent diagnostic cerebral angiography via R-TRA. Two of the patients received interventions via R-TRA: 1 underwent right internal carotid artery balloon dilation angioplasty, while another underwent left vertebral artery stenting. No surgery-related complications were observed during these procedures. CONCLUSIONS: R-TRA proves to be a safe and effective option for neuro-interventional surgery in patients with ARSA.

Minimally invasive spine surgery strategy for congenital cervicothoracic scoliosis in children: Less blood loss and shortened segmental fusions/fewer pedical screws.

Objective: To explore the feasibility of a minimally invasive spine surgery strategy for congenital cervicothoracic scoliosis. Materials and methods: From April 2022 to August 2022 in the hospital, three patients with torticollis and/or shoulder imbalance due to a cervicothoracic hemivertebra were performed on by hemivertebra resection and short fusion of the adjacent vertebrae. Resection was operated by a posterior approach. The average age of three patients of surgery was 8 years 2 months and the mean follow-up period was 6 months. Radiographic assessments and cosmetic outcomes were documented on changes in measurements of segmental scoliosis, neck tilt, head shift, shoulder balance, and sagittal profiles. Results: The mean operating time of the procedure was 283 min and the instrumentation density was 1.5 pedicle screws per vertebra. The mean estimated blood loss was 257 ml, which was 20% less than the data described in various literatures. The mean segmental Cobb angle at the cervicothoracic deformity was 35.9° before surgery, 20.7° after surgery, and 16.3° at the latest follow-up, with a correction rate of 54.59%. Neck tilt decreased from 17.3° before surgery to 14.3° after surgery, and 11.7° at the latest follow-up, with a correction rate of 32.37%. T1 tilt improved from 16.5° before surgery to 12.9° after surgery, and 7.6° at the latest follow-up, with a correction rate of 53.94%. The clavicle angle improved from 4.8° before surgery to 3.1° after surgery, and 1.9° at the latest follow-up, with a correction rate of 60.42%. Head shift improved from 21.4 mm before surgery to 9.2 mm after surgery, and 12.3 mm at the latest follow-up, with a correction rate of 42.52%. The correction of torticollis and shoulder asymmetry was achieved in all cases. Conclusions: Minimally invasive spine surgery strategy may be an option for congenital cervicothoracic scoliosis. A good correction of cervicothoracic dissymmetry is achieved, accompanied by fewer pedicle screws and less blood loss. By deliberate operation in young kids, surgical intervention for severe compensatory curves can be prevented.

SENP1 in the retrosplenial agranular cortex regulates core autistic-like symptoms in mice.

Autism spectrum disorder (ASD) is a highly heritable neurodevelopmental disorder, causing defects of social interaction and repetitive behaviors. Here, we identify a de novo heterozygous gene-truncating mutation of the Sentrin-specific peptidase1 (SENP1) gene in people with ASD without neurodevelopmental delay. We find that Senp1+/- mice exhibit core autistic-like symptoms such as social deficits and repetitive behaviors but normal learning and memory ability. Moreover, we find that inhibitory and excitatory synaptic functions are severely affected in the retrosplenial agranular (RSA) cortex of Senp1+/- mice. Lack of Senp1 leads to increased SUMOylation and degradation of fragile X mental retardation protein (FMRP), also implicated in syndromic ASD. Importantly, re-introducing SENP1 or FMRP specifically in RSA fully rescues the defects of synaptic function and autistic-like symptoms of Senp1+/- mice. Together, these results demonstrate that disruption of the SENP1-FMRP regulatory axis in the RSA causes autistic symptoms, providing a candidate region for ASD pathophysiology.

Smoothened is a therapeutic target for reducing glutamate toxicity in ischemic stroke.

Extracellular glutamate contributes to brain damage in ischemia. Under physiological conditions, glutamate transporters are responsible for regulating its intracellular/extracellular concentrations in the brain. However, how the extracellular glutamate is regulated in ischemia remains unclear. Here, we showed that the sonic hedgehog (SHH)­Smoothened (SMO)­GLT-1 pathway controlled extracellular glutamate and blocking SMO reduced ischemic brain damage in rodents. SHH was quickly released in a rodent model of ischemia, and activation of its pathway was associated with neuronal damage. Inhibiting SMO, the mediator of SHH signaling, maintained GLT-1 membrane expression, lowered extracellular glutamate, reduced infarct volume, and improved neurological functions in mice. Mechanistically, SHH suppressed GLT-1 membrane expression via PKCα phosphorylation of serine-562 on GLT-1. Last, administration of NVP-LDE225, an FDA-approved SMO antagonist used for cancer treatment in clinic, had protective effects in mice and cynomolgus monkeys subjected to ischemia. Together, these results suggest that SMO could be targeted for treating glutamate toxicity in ischemia.

Assessment of Patellar Morphology in Trochlear Dysplasia on Computed Tomography Scans.

OBJECTIVE: To evaluate the patellar morphology of trochlear dysplasia and normal knees in different genders and in different severities of trochlear dysplasia on CT scans. METHODS: A total of 75 patients with trochlear dysplasia (110 knees) treated at the Third Hospital of Hebei Medical University from December 2013 to December 2018 were included in an experimental group, and an age-matched and sex-matched cohort of 46 patients with normal trochlear shape (61 knees) were randomly selected into a control group. The experimental group was divided into a female experimental group (Group FE, 47 patients, 72 knees) and a male experimental group (Group ME, 28 patients, 38 knees); the control group was divided into a female control group (Group FC, 31 knees, 24 female patients) and a male control group (Group MC, 30 knees, 22 male patients). Furthermore, according to the severity of trochlear dysplasia, Group FE was divided into a female low-grade dysplasia group (Group FL, 20 knees) and a female high-grade dysplasia group (Group FH, 52 knees); Group ME was divided into a male low-grade dysplasia group (Group ML, 16 knees) and a male high-grade dysplasia group (Group MH, 22 knees). All participants had undergone CT scans in the supine position; the patellar width and thickness, the lateral patellar facet angle, the Wiberg angle, and the Wiberg index were measured and compared. RESULTS: In trochlear dysplasia knees, the mean patellar width and thickness and the lateral patellar facet angle were significantly smaller; the mean Wiberg index was significantly larger than in normal knees, regardless of gender (P < 0.05); and there was no statistically significant difference in the mean Wiberg angle (P > 0.05). In the female groups, the mean patellar width and thickness and the Wiberg angle were significantly smaller; the mean lateral patellar facet angle was significantly larger than those in the male groups (P < 0.05); and there was no significant difference in the mean Wiberg index (P > 0.05). In the low-grade dysplasia group, the mean Wiberg index was smaller than that in the high-grade dysplasia group (P < 0.05), regardless of gender; however, there was no significant difference in the mean patellar width and thickness, the lateral patellar facet angle, and the Wiberg angle in low-grade and high-grade dysplasia (P > 0.05). CONCLUSION: On CT scans, the patella in trochlear dysplasia had a smaller width, a thinner thickness, a lengthened lateral facet, and a more flattened articular facet. In addition, the patellar articular facet was more prominent in female patients. With the severity of trochlear dysplasia increased, the lateral patellar facet became longer. In addition, the abnormal stress distribution on the patella influenced the patellar morphology in trochlear dysaplasia.

iPhone-imaged and cell-powered electrophoresis titration chip for the alkaline phosphatase assay in serum by the moving reaction boundary.

As a vital enzyme, alkaline phosphatase (ALP) has great clinical significance in diagnoses of bone or liver cancer, bone metastases, rickets, and extrahepatic biliary obstruction. However, there is still no really portable chip for the ALP assay in blood. Herein, a simple electrophoresis titration (ET) model was developed for ALP detection via a moving reaction boundary (MRB). In the model, ALP catalyzed the dephosphorylation of a 4-methylumbelliferyl phosphate disodium salt (4-MUP) substrate in the cathode well to 4-methylumbelliferone ([4-MU]-) with a negative charge and blue fluorescence under UV excitation. After the catalysis, an electric field was used between the cathode and the anode. Under the electric field, [4-MU]- moved into the channel and neutralized the acidic Tris-HCl buffer, resulting in the quenching of [4-MU]- and creating a MRB. The ET system just had an ET chip, a lithium cell, a UV LED and an iPhone used as a recorder, having no traditional expensive power supply and fluorescence detector. The relevant method was developed, and a series of experiments were conducted via the ET chip. The experiments showed: (i) a MRB could be formed between the [4-MU]- base and the acidic buffer, and the MRB motion had a linear relationship with the ALP activity, validating the ET model; (ii) the ET run was not impacted by many interferences, implying good selectivity; and (iii) the ET chip could be used for portable detection within 10 min, implying an on-site and rapid analysis. In addition, the ET method had a relatively good sensitivity (0.1 U L-1), linearity (V = 0.033A + 3.87, R2 = 0.9980), stability (RSD 2.4-6.8%) and recoveries (101-105%). Finally, the ET method was successfully used for ALP assays in real serum samples. All the results implied that the developed method was simple, rapid and low-cost, and had potential for POCT clinical ALP assays.

Graphene Oxide-Facilitated Comprehensive Analysis of Cellular Nucleic Acid Binding Proteins for Lung Cancer.

Nucleic acid binding proteins (NABPs) mediate a broad range of essential cellular functions. However, it is very challenging to comprehensively extract whole cellular NABPs due to the lack of approaches with high efficiency. To this end, carbon nanomaterials, including graphene oxide (GO), carboxylated graphene (cG), and carboxylated carbon nanotube (cCNT), were utilized to extract cellular NABPs in this study through a new strategy. Our data demonstrated that GO, cG, and cCNT could extract nearly 100% cellular DNA in vitro. Conversely, their RNA extraction efficiencies were 60, 50, and 29%, respectively, partially explaining why GO has the highest NABPs yield compared to cG and cCNT. We further found that ionic bond mediated by cations between RNA and functional groups of nanomaterials facilitated RNA absorption on nanomaterials. About 2400 proteins were successfully identified from GO-enriched NABPs sample, and 88% of annotated NABPs were enriched at least 2 times compared to cell lysate, indicating the high selectivity of our strategy. The developed method was further applied to compare the NABPs in two lung cancer cell lines with different tumor progression abilities. According to label-free quantification results, 118 differentially expressed NABPs were discovered and 6 candidate NABPs, including ACAA2, GTF2I, VIM, SAMHD1, LYAR, and IGF2BP1, were successfully validated by immunoassay. The level of SAMHD1 in the serum of lung cancer patients was measured, which significantly increased upon cancer progression. Our results collectively demonstrated that GO is an ideal nanomaterial for NABPs selective extraction, which could be broadly used in varied physiological and pathophysiological settings.

Comparative Proteomic Analysis of Exosomes and Microvesicles in Human Saliva for Lung Cancer.

Extracellular vesicles (EVs) are cell-derived microparticles present in most body fluids, mainly including microvesicles and exosomes. EV-harbored proteins have emerged as novel biomarkers for the diagnosis and prediction of different cancers. We successfully isolated microvesicles and exosomes from human saliva, which were further characterized comprehensively. Salivary EV protein profiling in normal subjects and lung cancer patients was systematically compared through utilizing LC-MS/MS-based label-free quantification. 785 and 910 proteins were identified from salivary exosomes and microvesicles, respectively. According to statistical analysis, 150 and 243 proteins were revealed as dysregulated candidates in exosomes and microvesicles for lung cancer. Among them, 25 and 40 proteins originally from distal organ cells were found in the salivary exosomes and microvesicles of lung cancer patients. In particular, 5 out of 25 and 9 out of 40 are lung-related proteins. Six potential candidates were selected for verification by Western blot, and four of them, namely, BPIFA1, CRNN, MUC5B, and IQGAP, were confirmed either in salivary microvesicles or in exosomes. Our data collectively demonstrate that salivary EVs harbor informative proteins that might be used for the detection of lung cancer through a noninvasive way.

Sonic hedgehog is a regulator of extracellular glutamate levels and epilepsy.

Sonic hedgehog (Shh), both as a mitogen and as a morphogen, plays an important role in cell proliferation and differentiation during early development. Here, we show that Shh inhibits glutamate transporter activities in neurons, rapidly enhances extracellular glutamate levels, and affects the development of epilepsy. Shh is quickly released in response to epileptic, but not physiological, stimuli. Inhibition of neuronal glutamate transporters by Shh depends on heterotrimeric G protein subunit Gαi and enhances extracellular glutamate levels. Inhibiting Shh signaling greatly reduces epileptiform activities in both cell cultures and hippocampal slices. Moreover, pharmacological or genetic inhibition of Shh signaling markedly suppresses epileptic phenotypes in kindling or pilocarpine models. Our results suggest that Shh contributes to the development of epilepsy and suppression of its signaling prevents the development of the disease. Thus, Shh can act as a modulator of neuronal activity, rapidly regulating glutamate levels and promoting epilepsy.

Facile preparation of salivary extracellular vesicles for cancer proteomics.

Extracellular vesicles (EVs) are membrane surrounded structures released by cells, which have been increasingly recognized as mediators of intercellular communication. Recent reports indicate that EVs participate in important biological processes and could serve as potential source for cancer biomarkers. As an attractive EVs source with merit of non-invasiveness, human saliva is a unique medium for clinical diagnostics. Thus, we proposed a facile approach to prepare salivary extracellular vesicles (SEVs). Affinity chromatography column combined with filter system (ACCF) was developed to efficiently remove the high abundant proteins and viscous interferences of saliva. Protein profiling in the SEVs obtained by this strategy was compared with conventional centrifugation method, which demonstrated that about 70% more SEVs proteins could be revealed. To explore its utility for cancer proteomics, we analyzed the proteome of SEVs in lung cancer patients and normal controls. Shotgun proteomic analysis illustrated that 113 and 95 proteins have been identified in cancer group and control group, respectively. Among those 63 proteins that have been consistently discovered only in cancer group, 12 proteins are lung cancer related. Our results demonstrated that SEVs prepared through the developed strategy are valuable samples for proteomics and could serve as a promising liquid biopsy for cancer.

Differential Proteomic Analysis of Human Saliva using Tandem Mass Tags Quantification for Gastric Cancer Detection.

Novel biomarkers and non-invasive diagnostic methods are urgently needed for the screening of gastric cancer to reduce its high mortality. We employed quantitative proteomics approach to develop discriminatory biomarker signatures from human saliva for the detection of gastric cancer. Salivary proteins were analyzed and compared between gastric cancer patients and matched control subjects by using tandem mass tags (TMT) technology. More than 500 proteins were identified with quantification, and 48 of them showed significant difference expression (p < 0.05) between normal controls and gastric cancer patients, including 7 up-regulated proteins and 41 down-regulated proteins. Five proteins were selected for initial verification by ELISA and three were successfully verified, namely cystatin B (CSTB), triosephosphate isomerase (TPI1), and deleted in malignant brain tumors 1 protein (DMBT1). All three proteins could differentiate gastric cancer patients from normal control subjects, dramatically (p < 0.05). The combination of these three biomarkers could reach 85% sensitivity and 80% specificity for the detection of gastric cancer with accuracy of 0.93. This study provides the proof of concept of salivary biomarkers for the non-invasive detection of gastric cancer. It is highly encouraging to turn these biomarkers into an applicable clinical test after large scale validation.

A tunable isoelectric focusing via moving reaction boundary for two-dimensional gel electrophoresis and proteomics.

Routine native immobilized pH gradient isoelectric focusing (IPG-IEF) and two-dimensional gel electrophoresis (2DE) are still suffering from unfortunate reproducibility, poor resolution (caused by protein precipitation) and instability in characterization of intact protein isoforms and posttranslational modifications. Based on the concept of moving reaction boundary (MRB), we firstly proposed a tunable non-IPG-IEF system to address these issues. By choosing proper pairs of catholyte and anolyte, we could achieve desired cathodic and anodic migrating pH gradients in non-IPG-IEF system, effectively eliminating protein precipitation and uncertainty of quantitation existing in routine IEF and 2DE, and enhancing the resolution and sensitivity of IEF. Then, an adjustable 2DE system was developed by combining non-IPG-IEF with polyacrylamide gel electrophoresis (PAGE). The improved 2DE was evaluated by testing model proteins and colon cancer cell lysates. The experiments revealed that (i) a tunable pH gradient could be designed via MRB; (ii) up to 1.65 fold improvement of resolution was achieved via non-IPG-IEF; (iii) the sensitivity of developed techniques was increased up to 2.7 folds; and (iv) up to about 16.4% more protein spots could be observed via the adjustable 2DE as compared with routine one. The developed techniques might contribute to complex proteome research, especially for screening of biological marker and analysis of extreme acidic/alkaline proteins.

A new strategy for highly efficient single-drop microextraction with a liquid-gas compound pendant drop.

Herein, a simple assembly was designed via a capillary and a funnel-like cap to achieve liquid-gas compound pendant drop (CPD) microextraction with great convenience. Due to the increased contact area and adhesion force between the capillary tip and the drop, the proposed method provides considerable flexibility in producing CPDs with different air bubble sizes. Four pesticides were chosen as model analytes to evaluate the proposed method. By using a 1 µL chlorobenzene droplet containing a 1 µL air bubble at a stirring rate of 700 rpm, a 70 to 135-fold enrichment of pesticides was obtained within 3.4 minutes. As compared with a typical SDME, the proposed method showed a 2-fold increase of enrichment factors and a 4-fold decrease of extraction time. Improvement of the extraction efficiency could be ascribed to the increased surface area of the droplet, and the thin film phenomena further improved the extraction kinetics through effective agitation. The results indicate that CPD microextraction could serve as a promising sample pretreatment method for automated high-throughput analyses in a wide variety of research areas.

Inhibition of Six1 promotes apoptosis, suppresses proliferation, and migration of osteosarcoma cells.

Sineoculis homeobox homolog 1 (Six1) is one of the transcription factors that act as master regulators of development and is frequently dysregulated in cancers. However, the biological role of Six1 is not clear in osteosarcoma. To address the expression of Six1 in osteosarcoma cells, three osteosarcoma cell lines (U2OS, SaOS-2, and MG63) and a human osteoblastic cell line (hFOB1.19) were used to detect the expression of Six1 by quantitative real-time polymerase chain reaction and western blotting. The results showed that Six1 was upregulated in osteosarcoma cell lines compared to human osteoblastic cell line hFOB1.19. To investigate the role of Six1 in osteosarcoma cells, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry analysis, and transwell chamber assays were used to determine the effects of Six1 on the cell viability, cycle, apoptosis, and migration properties in U2OS cells. The results showed that Six1 could promote U2OS cell proliferation and migration, and suppress U2OS cell apoptosis. In addition, we investigated the effects of Six1 on the expression of following proteins (cyclin D1, caspase-3, and vascular endothelial growth factor-C (VEGF-C)). Results showed that Six1 could increase the expression of cyclin D1 and VEGF-C, and decrease the expression of caspase-3. All these data suggested that Six1 might be involved in the promotion of growth, proliferation, and migration of U2OS cells, as well as the inhibition of apoptosis of U2OS cells. These data might provide information for the prediction of osteosarcoma prognosis and potential targets for therapy of osteosarcoma.

Impact of glutathione-HbA1c on HbA1c measurement in diabetes diagnosis via array isoelectric focusing, liquid chromatography, mass spectrometry and ELISA.

Hemoglobin A1c (HbA1c) has been proven to be a key biomarker for diabetes screening, and glutathiolation of HbA1c (viz., GSS-HbA1c) has been identified. However, the impact of GSS-HbA1c on the measurement of HbA1c for diabetes screening has not been quantitatively assessed yet. To address the issue, the micropreparative capillary isoelectric focusing (cIEF) developed in our previous work was used for the high resolution separation and purification of hemoglobin (Hb) species. The main fractions of HbA0, HbA3 and HbA1c extracted from the developed cIEF were identified by validated Mono S method. The proposed GSS-HbA1c fractions in the cIEF were pooled and identified by electrospray ionization mass spectrometry (ESI-MS). The HbA1c enzyme-linked immunosorbent assay (ELISA) kit was employed for further quantitative analysis of GSS-HbA1c. A total of 34 blood samples with HbA1c levels from 4.2% to 13.4% were assessed via the above comprehensive strategy of IEF-HPLC-MS-ELISA. It was demonstrated that the HbA1c levels detected by cation exchange LC were considerably influenced by the glutathiolation of Hb and the range of detected GSS-HbA1c values was between 0.23% and 0.74%. The results and developed cIEF methods have considerable significances for investigation of diabetes and clinical diagnosis.

MicroRNA 181a improves proliferation and invasion, suppresses apoptosis of osteosarcoma cell.

MicroRNA 181a (miR-181a) was found dysregulated in a variety of human cancers and significantly associated with clinical outcome of cancer patients. However, the direct role of miR-181a has not yet been characterized in osteosarcoma progression. This study was aimed at investigating the effects of miR-181a on osteosarcoma cell biological behavior. First, the expression of miR-181a in osteosarcoma cell lines (MG63, HOS, SaOS-2, and U2OS) and a human osteoblastic cell line (hFOB1.19) was detected by qRT-PCR. Results showed that miR-181a was overexpressed in osteosarcoma cell lines compared to human osteoblastic cell line (hFOB1.19). To investigate the effects of miR-181a on proliferation, apoptosis, and invasion of osteosarcoma cells, we generated human osteosarcoma MG63 cells in which miR-181a was either overexpressed or depleted. The MG63 cell viability, cycle, apoptosis, and invasive ability were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide staining, propidium iodide (PI) staining, Annexin V-FITC/PI double staining, and Transwell invasion experiment, respectively. The results showed that MG63 cell viability, proliferation, and invasive abilities were suppressed, and the apoptosis was enhanced in the group with underexpression of miR-181a. The viability, proliferation, and invasive abilities were improved, and the apoptosis was inhibited in the group with overexpression of miR-181a. The results from Western blotting indicated that miR-181a might be associated with the up-regulation of bcl-2 and matrix metalloproteinase 9 and the down-regulation of tissue inhibitor of metalloproteinases-3 and p21 in MG63 cells. Taken together, our results suggested that miR-181a might facilitate proliferation and invasion and suppress apoptosis of osteosarcoma cells, which might be a potential target for the treatment of osteosarcoma.

Effects of Kruppel-like factor 6 on osteosarcoma cell biological behavior.

Kruppel-like factor 6 (KLF6) is a tumor suppressor gene frequently downregulated in a number of human cancers, including osteosarcoma. However, the role of KLF6 in osteosarcoma remains unclear. This study was aimed at investigating the effects of KLF6 on osteosarcoma cell biological behavior. First, the expression of KLF6 in osteosarcoma cell lines (MG63, SaOS-2, U2OS, and HOS) and a human osteoblastic cell line (hFOB1.19) was detected by Western blotting. Results showed that KLF6 displayed a significant downregulation in osteosarcoma cell lines (MG63, SaOS-2, U2OS, and HOS) compared with human osteoblastic cell line (hFOB1.19). To investigate the role of KLF6 in osteosarcoma cell proliferation, apoptosis, and invasion, we generated human osteosarcoma MG63 cells in which KLF6 was either overexpressed or depleted. The MG63 cell viability, cycle, apoptosis, and invasive ability were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide staining, propidium iodide (PI) staining, Annexin-V-FITC/PI double staining, and Transwell invasion experiment, respectively. Results showed that the viability, proliferation, and invasive abilities were suppressed, and the apoptosis was enhanced in MG63 cells with overexpression of KLF6. The viability, proliferation, and invasive abilities were improved, and the apoptosis was inhibited in MG63 cells with knockdown of KLF6. At the same time, these molecules, including p21, bcl-2, and MMP-9, associated with the events about cell cycle, apoptosis, and invasion, were detected. Results showed that the expressions of bcl-2 and MMP-9 were downregulated, and the expressions of p21 were upregulated in the MG-63 cells with overexpression of KLF6. Taken together, our results suggested that KLF6 could inhibit proliferation and invasion, and facilitate apoptosis of osteosarcoma cells, which might be a potential target for the treatment of osteosarcoma.

Separation and determination of four ganoderic acids from dried fermentation mycelia powder of Ganoderma lucidum by capillary zone electrophoresis.

Ganoderic acids (GAs) were bioactive secondary metabolites produced by a traditional mushroom Ganoderma lucidum. We describe a simple and efficient method for the separation and quantitative determination of four GAs, namely Ganoderic acid T (GA-T), Ganoderic acid Mk (GA-Mk), Ganoderic acid Me (GA-Me) and Ganoderic acid S (GA-S) from dried triterpene-enriched extracts of G. lucidum mycelia powder by capillary zone electrophoresis (CZE). Under the optimum conditions, the four GAs reached the baseline separation in 9 min with Glycyrrhetinic acid (GTA) as internal standard. The four GAs and internal standard (GTA) were detected at a wavelength 245 nm. All calibration curves showed good linearity (r(2)>0.9958) within test ranges. Limit of detection (LOD) and limit of quantification (LOQ) were less than 0.6 and 1.8 microg/mL, respectively. The relative standard deviation (R.S.D.) values of precision and recoveries were less than 5% and recoveries ranged from 91.4% to 103.6%. This was the first report on simultaneous determination of the four GAs and the results provided a firm basis for the trace analysis of GAs in dried fermentation mycelia powder of G. lucidum with high accuracy.

A simple preparative free-flow electrophoresis joined with gratis gravity: I. Gas cushion injector and self-balance collector instead of multiple channel pump.

This paper describes a novel free-flow electrophoresis (FFE), which is joined with gratis gravity, gas cushion injector (GCI) and self-balance collector instead of multiple channel pump, for the purpose of preparative purification. The FFE was evaluated by systemic experiments. The results manifest that (i) even though one-channel peristaltic pump is used for the driving of background buffer, there is still stable flow in the FFE chamber; (ii) the stable flow is induced by the gravity-induced pressure due to the difference of buffer surfaces in the GCI and self-balance collector; (iii) the pulse flow of background buffer induced by the peristaltic pump is greatly reduced by the GCI with good compressibility of included air; (iv) the FFE can be well used for zone electrophoretic separation of amino acids; (v) up to 20 inlets simultaneous sample injection and up to five to tenfold condensation of amino acid can be achieved by combining the FFE device with the method of moving reaction boundary. To the best of authors' knowledge, FFE has not been used for such separation and condensation of amino acids. The relevant results achieved in the paper have evident significance for the development of preparative FFE.

[Clinical application value of laryngeal electromyography and laryngeal somatosensory evoked potential in patients with recurrent laryngeal nerve paralysis by anterior elective cervical surgery].

OBJECTIVE: To determine the clinical evaluation role of laryngeal electromyography (LEMG) and laryngeal somatosensory evoked potential (LSEP) in the recurrent laryngeal nerve paralysis by anterior elective cervical surgery. METHODS: LEMG and LSEP were determined in 18 patients with recurrent laryngeal nerve paralysis by anterior elective cervical surgery at the 1st, 2nd and 4th week after operation. The comparison between the normal control (18 health adults) and the results of LEMG and LSEP were analyzed. RESULTS: The latency prolonged and the amplitude decreased of LSEP in all patients as compared with the control group. Furthermore, reinneration potential increased gradually in all patients at the 1st, 2nd and 4th week after operation (P<0.05). The results of LEMG showed increase of denervation potential. The higher the amplitude of LSEP and LEMG, the better the prognosis of the recurrent laryngeal nerve paralysis. CONCLUSION: LEMG and LSEP might evaluate regenerate the degree of recurrent laryngeal nerve injury caused by anterior elective cervical surgery and predict the prognosis of the recurrent laryngeal nerve paralysis.