Insight into the compositional variation of volatile and polar compounds of Radix Bupleuri from different processing technologies by GC-MS and UHPLC-Q-TOF/MS metabolomics.

INTRODUCTION: Insight into comparing key active ingredients of Radix Bupleuri (RB) based on different processing technologies is a key step to reveal the material basis of drug efficacy and a challenging task for developing traditional Chinese medicine (TCM). OBJECTIVE: This work aims to establish a comprehensive comparative analysis method of TCM and its processed products, which can be used to analyze the changing trend of active components of RB before and after processing. METHODS: First, RB was processed with rice vinegar, rice wine, and honey. Then, ultra-high-performance liquid chromatography (UHPLC) and gas chromatography (GC) coupled with mass spectrometry (MS) technology as well as multiple statistical analyses were used to comprehensively evaluate the compositional variation of polar and volatile compounds in RB under different processing processes. Meanwhile, in UHPLC-MS, a sequential window acquisition of all theoretical fragment ion spectral and information-dependent acquisition mutual authentication (SIMA) was developed. RESULTS: A total of 30 polar components and 33 volatile components were identified as chemical markers (mainly type II saikosaponins, terpenes, and fatty acid esters). These may be the material basis for giving unique pharmacological activities to RB and its processed products. CONCLUSIONS: These findings provided a solid foundation for the differentiated clinical application of RB, and the SIMA method held great potential for achieving accurate analysis of TCM processing ingredients.

The Role of CT Perfusion in Differentiating Benign Versus Malignant Focal Pulmonary Lesions.

BACKGROUND: Contrast-enhanced CT scan is the standard imaging for the characterization and evaluation of focal parenchymal lung lesions. It relies on morphology and enhancement patterns for the characterization of lung lesions. However, there is significant overlap among imaging features of various malignant and benign lesions. Hence, it is often necessary to obtain tissue diagnosis with invasive percutaneous or endoscopic-guided tissue sampling. It is often desirable to have non-invasive techniques that can differentiate malignant and benign lung lesions. CT perfusion is an emerging CT technology that allows functional assessment of tissue vascularity through various parameters and can help in differentiating benign and malignant focal lung lesions. OBJECTIVE: The purpose of this study was to assess the role of the CT perfusion technique in differentiating malignant and benign focal parenchymal lung lesions. MATERIALS AND METHODS: In this prospective observational study, CT perfusion was performed on 41 patients with focal parenchymal lung lesions from December 2020 to June 2022. The four-dimensional range was planned to cover the entire craniocaudal extent of the lesion, followed by a volume perfusion CT (VPCT) of the lesion. A total of 27 dynamic datasets were acquired with a scan interval of 1.5 seconds and a total scan time of 42 seconds. CT perfusion parameters of blood flow (BF), blood volume (BV), and k-trans of the lesion were measured with mathematical algorithms available in the Syngo.via CT perfusion software (Siemens Healthcare, Erlangen, Germany). RESULTS: The median BV in benign lesions was found to be 5.5 mL/100 g, with an interquartile range of 3.3-6.9 and a p-value < 0.001. The median BV in malignant lesions was found to be 11.35 mL/100 g, with an interquartile range of 9.57-13.21 and a p-value ≤ 0.001. The median BF for benign lesions was 45.5 mL/100 g/min, with an interquartile range of 33.8-48.5 and a p-value ≤ 0.001. The median BF for malignant lesion was 61.77 mL/100 g/min, with an interquartile range of 33.8-48.5 and a p-value ≤ 0.001. The median k-trans in the case of benign lesions was found to be 4.2 mL/100 g/min, with an interquartile range of 3.13-6.8 and a p-value ≤ 0.001. The median k-trans in the case of the malignant lesion was found to be 12.05 mL/100g/min, with an interquartile range of 7.20-33.42 and a p-value < 0.001. Our study has also shown BV to have an accuracy of 92.68%, sensitivity of 93.3%, and specificity of 90.01%. CONCLUSION: Our study has shown that CT perfusion values of BV, BF, and k-trans can be used to differentiate between benign and malignant focal lung parenchymal lesions. K-trans is the most sensitive parameter while BV and BF have greater accuracy and specificity.

Human-mouse chimeric brain models constructed from iPSC-derived brain cells: Applications and challenges.

The establishment of reliable human brain models is pivotal for elucidating specific disease mechanisms and facilitating the discovery of novel therapeutic strategies for human brain disorders. Human induced pluripotent stem cell (iPSC) exhibit remarkable self-renewal capabilities and can differentiate into specialized cell types. This makes them a valuable cell source for xenogeneic or allogeneic transplantation. Human-mouse chimeric brain models constructed from iPSC-derived brain cells have emerged as valuable tools for modeling human brain diseases and exploring potential therapeutic strategies for brain disorders. Moreover, the integration and functionality of grafted stem cells has been effectively assessed using these models. Therefore, this review provides a comprehensive overview of recent progress in differentiating human iPSC into various highly specialized types of brain cells. This review evaluates the characteristics and functions of the human-mouse chimeric brain model. We highlight its potential roles in brain function and its ability to reconstruct neural circuitry in vivo. Additionally, we elucidate factors that influence the integration and differentiation of human iPSC-derived brain cells in vivo. This review further sought to provide suitable research models for cell transplantation therapy. These research models provide new insights into neuropsychiatric disorders, infectious diseases, and brain injuries, thereby advancing related clinical and academic research.

A novel and cost-effective real-time RT-PCR targeting 24 nucleotides deletion to differentiate PEDV wild-type and classical attenuated vaccine strains.

Porcine Epidemic Diarrhea Virus (PEDV) poses a significant threat to the swine industry, causing severe disease and resulting in substantial economic losses. Despite China's implementation of a large-scale vaccine immunization strategy in recent years, various strains of PEDV, including classical attenuated vaccine strains, continue to emerge in immunized pig herds. Here, we established a one-step real-time fluorescent reverse transcription PCR (one-step real-time RT-PCR) assay targeting a 24-nucleotide deletion in the ORF1 region of three PEDV classical attenuated vaccine strains, derived from classical strains. This assay effectively distinguishes between PEDV classical attenuated vaccine strains and wild-type strains, and we also explore the causes of this discriminatory target deficiency of this method through phylogenetic and recombination analysis. We found that these three classical attenuated vaccine strains exhibit closer phylogenetic relationships and higher sequence similarity with five cell-adapted strains. Recombination analysis revealed that although recombination is widespread in the PEDV genome, the 24-nucleotide deletion site remains stable without undergoing recombination and can be utilized as a target for identification. Further analysis revealed there are no enzyme cleavage sites near the 24-nucleotide site, suggesting that this deletion may have been lost during the process of culturing these viral strains in cells.The detection method we have established exhibits high specificity and sensitivity to PEDV, without cross-reactivity with other viruses causing diarrheal diseases. A total of 117 swine fecal samples were analyzed using this established one-step real-time reverse transcription PCR assay, indicating the presence of classical attenuated vaccine strains in pig herds in Gansu province, China. Additionally, the designed primer pairs and two probes can be placed in a single reaction tube to differentiate between these two types of strains, effectively reducing detection costs. These findings offer an efficient and cost-effective technological platform for clinical rapid identification testing of both wild-type and classical attenuated vaccine strains of PEDV, as well as for precise investigation of clinical data on natural infections and vaccine immunity in pig herds.

Culturing Immortalized Human Airway Epithelial Cells at an Air-Liquid Interface for Measles Virus Infection.

Measles virus (MeV) infection of airway surface epithelial cells provides a site for final amplification before being released back into the environment via coughing and sneezing. Multiple cell lines have served as models of polarized epithelia for MeV infection, such as Caco2 cells (intestinal derived human epithelia) or MDCK cells (kidney derived canine epithelia). In this chapter, we describe the materials and air-liquid interface (ALI) culture conditions for maintaining four different cell lines derived from human airway epithelial cells: 16HBE14o-, Calu-3, H358, and NuLi-1. We provide methods for confirming transepithelial electrical resistance (TER) and preparing samples for microscopy as well as expected results from apical or basolateral MeV delivery. Polarized human airway derived cells serve as tissue culture models for investigating targeted questions about how MeV exits a human host. In addition, these methods are generalizable to studies of other respiratory viruses or the biology of ALI airway epithelial cells.

Ultrasound-Based Predictive Model to Assess the Risk of Orbital Malignancies.

OBJECTIVE: Ultrasound (US) is widely used for evaluating various orbital conditions. However, accurately diagnosing malignant orbital masses using US remains challenging. We aimed to develop an ultrasonic feature-based model to predict the presence of malignant tumors in the orbit. METHODS: A total of 510 patients with orbital masses were enrolled between January 2017 and April 2023. They were divided into a development cohort and a validation cohort. In the development cohort (n = 408), the ultrasonic and clinical features with differential values were identified. Based on these features, a predictive model and nomogram were constructed. The diagnostic performance of the model was compared with that of MRI or observers, and further validated in the validation cohort (n = 102). RESULTS: The involvement of more than two quadrants, irregular shape, extremely low echo of the solid part, presence of echogenic foci, cast-like appearance, and two demographic characteristics (age and sex) were identified as independent features related to malignant tumors of the orbit. The predictive model constructed based on these features exhibited better performance in identifying malignant tumors compared to MRI (AUC = 0.78 [95% CI: 0.73, 0.82] vs. 0.69 [95% CI: 0.64, 0.74], p = 0.03) and observers (AUC = 0.93 [95% CI: 0.90, 0.95] vs. Observer 1, AUC = 0.80 [95% CI: 0.76, 0.84], p < 0.01; vs. Observer 2, AUC = 0.71 [95% CI: 0.66, 0.76], p < 0.01). In the validation cohort, the predictive model achieved an AUC of 0.88 (95% CI: 0.81, 0.94). CONCLUSION: The ultrasonic-clinical feature-based predictive model can accurately identify malignant orbital tumors, offering a convenient approach in clinical practice.

Can99mTc-MDP-SPECT/CT Differentiate Loosening and Infection After Hip and Knee Replacements?

Background: Prosthetic loosening and infection are still common complications after joint replacement. Over the past few years, single-photon emission computed tomography-computed tomography (SPECT/CT) was widely used and showed unique value based on the combination of anatomic and metabolic information of foci. However, its performance in differentiating between prosthetic loosening and periprosthetic infection after joint replacement is still the focus of clinicians and deserves further investigation. Purpose: This retrospective study was aimed to determine whether bone scintigraphy combined with SPECT/CT still can differentiate prosthetic infection from loosening in patients after joint replacement. The differential efficacy in hip and knee prosthesis was also analyzed. Blood biomarkers for the diagnosis of periprosthetic infection were also evaluated. Patients and methods: Data sets of 74 prosthetic joints (including knees and hips), with suspected prosthetic loosening or infection between 2015 and 2021, were evaluated. Besides the results of nuclear imaging, X-ray images and serum biomarker were also recorded. Telephone follow-up and revision surgery after SPECT/CT were used as a gold standard. The sensitivity and accuracy of different imaging modalities were calculated by Chi-square test. The diagnostic efficacy of imaging methods and serum biomarkers were then analyzed by the area under curve (receiver operating characteristic curves, ROC) in SPSS 26. Results: In all, 47 joints (14 knees and 33 hips) were confirmed as aseptic loosening, while 25 joints (18 knees and 7 hips) were confirmed as infection. The sensitivity and accuracy of SPECT combined with SPECT/CT imaging were the highest (92.86% and 87.84%, respectively). The differential efficacy of bone scintigraphy combined with SPECT/CT imaging was also better than any other single imaging modality. In the analysis of involved prosthesis, prosthetic loosening occurred more in hip prosthesis and knee prosthesis was easily infected (P < 0.05). Finally, the sensitivity of ESR and CRP were 80% and 84%, respectively. Conclusions: Bone scintigraphy with hybrid SPECT/CT remains encouraging in differentiating prosthetic infection from loosening after joint replacement. The diagnostic efficacy of differentiation in hip prosthesis was better than knee. Serum biomarkers cannot be used alone to differentiate prosthetic infection from loosening.

A Prospective Study Between Molecular Marker P53 Analysis and HPE, for Tumour Positivity in Primary Resected Margins in Head and Neck Squamous Cell Carcinoma.

Head and neck malignancies are the seventeenth most common malignancies, worldwide and second most common malignancy in India. So current study aims to compare between molecular marker p53 analysis and HPE for tumour positivity in primary resected margins in head and neck SCC. 61 patients with head and neck SCC were included in this prospective observational cross-sectional study performed in tertiary care hospital. A detailed history general physical examination blood investigation was done before the surgery. After the surgery, primary lesion from the resected tumour was sent for HPE analysis. From the same specimen, the margins at distance of 0.5-1 cm from primary tumour were sent for p53 mutation analysis. Report of p-53 mutation was noted and entered to the Performa. In our study we found out that in PDSCC HPE negative margins were found positive for p53 mutation in 81% cases. Which suggest that evaluation for p53 mutation should be done in PDSCC cases for HPE negative margins with in 1 cm. In patients of head and neck squamous cell carcinoma with free margins on HPE p-53 mutation is significantly associated to the PDSCC and margin upto 0.7 mm so recommended for p-53 profile can be beneficial in cases of the PDSCC and margins up to 0.7 mm for further management or for possibility of recurrence and its management to improve patients survival and decrease morbidity and mortality.

Expression of CD 20 B-Lymphocyte in oral epithelial dysplasia and oral squamous cell carcinoma: A comparative immunohistochemistry study.

Background: As the progressive trends in the field of immunotherapy, it is very favourable to reconsider the role played by B lymphocytes in the tumor microenvironment. Both the protumorogenic and antitumorogenic responses have to be evaluated to formulate an effective immunotherapeutic protocol. Aim and objective: The study was primarily conducted to assess the qualitative expression of B lymphocytes in pretumorogenic (oral epithelial dysplasia) and tumorogenic environment (oral squamous cell carcinoma). The differential immunohistochemical staining of CD 20 immune marker was assessed in about 60 cases that included 30 cases of oral epithelial dysplasia and 30 cases of oral squamous cell carcinoma. Results: The study found significant correlation between CD 20 IHC immune expression and histopathological diagnosis along with significant correlation between the subject's age group and histopathological diagnosis. Conclusion: Modulating the immune response in a precancerous state can be highly beneficial in implementing better immunotherapeutic strategies to treat or prevent malignancy at an early stage.

Representative measles virus infection requires appropriate airway epithelia culture conditions.

IMPORTANCE: For many years, measles virus (MeV) was assumed to first enter the host via the apical surface of airway epithelial cells and subsequently spread systemically. We and others reported that MeV has an overwhelming preference for entry at the basolateral surface of airway epithelial cells, which led to a fundamental new understanding of how MeV enters a human host. This unexpected observation using well-differentiated primary cultures of airway epithelia from human donors contradicted previous studies using immortalized cultured cells. Here, we show that appropriate differentiation and cell morphology of primary human airway epithelial cells are critical to recapitulate MeV infection patterns and pathogenesis of the in vivo airways. By simply culturing primary cells in media containing serum or passaging primary cultures, erroneous results quickly emerge. These results have broad implications for data interpretation related to respiratory virus infection, spread, and release from human airway epithelial cells.

Differentiating brain states via multi-clip random fragment strategy-based interactive bidirectional recurrent neural network.

EEG is widely adopted to study the brain and brain computer interface (BCI) for its non-invasiveness and low costs. Specifically EEG can be applied to differentiate brain states, which is important for better understanding the working mechanisms of the brain. Recurrent neural network (RNN)-based learning strategy has been widely utilized to differentiate brain states, because its optimization architectures improve the classification performance for differentiating brain states at the group level. However, present classification performance is still far from satisfactory. We have identified two major focal points for improvements: one is about organizing the input EEG signals, and the other is related to the design of the RNN architecture. To optimize the above-mentioned issues and achieve better brain state classification performance, we propose a novel multi-clip random fragment strategy-based interactive bidirectional recurrent neural network (McRFS-IBiRNN) model in this work. This model has two advantages over previous methods. First, the McRFS component is designed to re-organize the input EEG signals to make them more suitable for the RNN architecture. Second, the IBiRNN component is an innovative design to model the RNN layers with interaction connections to enhance the fusion of bidirectional features. By adopting the proposed model, promising brain states classification performances are obtained. For example, 96.97% and 99.34% of individual and group level four-category classification accuracies are successfully obtained on the EEG motor/imagery dataset, respectively. A 99.01% accuracy can be observed for four-category classification tasks with new subjects not seen before, which demonstrates the generalization of our proposed method. Compared with existing methods, our model outperforms them with superior results. Overall, the proposed McRFS-IBiRNN model demonstrates great superiority in differentiating brain states on EEG signals.

Outer membrane protein BamA-based ELISA differentiates Salmonella-vaccinated chickens from naturally infected chickens.

Salmonella often causes subclinical infection in chickens, but antibody tests can find infected individuals and control the spread of infection. In this study, the S. Typhimurium-specific outer membrane, ß-barrel assembly machinery protein A (BamA), was overexpressed in Escherichia coli and purified as a coating antigen to develop a BamA-based enzyme-linked immuno sorbent assay for detecting Salmonella infection. The presence of anti-BamA IgG was detected in the sera of infected BALB/c mice, but not in that of heat-killed Salmonella-vaccinated mice. The assay was validated using White Leghorn chickens and showed similar results. The detection of BamA antibodies in the sera can differentiate infected chickens from vaccinated chickens. This assay will be useful for monitoring Salmonella infection in chickens and possibly in other animals.

Development of trisiloxane surfactant vesicles ultrasonic extraction method combined with ultrahigh-performance liquid chromatography tandem mass spectrometry for the rapid differentiation of Bupleuri Radix based on metabolomics.

INTRODUCTION: Due to the variety, chemical composition and complex structure, the quality control of Bupleuri Radix (BR) is a challenging task. There are still many trace compounds in BR that are difficult to extract and detect. OBJECTIVE: To develop an innovative method of trisiloxane surfactant vesicles ultrasonic extraction (TSVUE) combined with ultrahigh-performance liquid chromatography tandem mass spectrometry for the identification from Bupleurum chinense DC. (BC) to Bupleurum scorzonerifolium Willd (BS) based on metabolomics. METHODS: Based on extraction effect for BR, five different types of surfactants vesicles were prepared and compared. Then, a single-factor test and a response surface methodology study were adopted to obtain the optimal conditions for the surfactant vesicles ultrasonic extraction method. Finally, a non-targeted metabolomics method with information dependent acquisition mode was performed to analyse differential metabolites in BC and BS. RESULTS: Sugar-based surfactant containing trisiloxane [N-3-propyl-methyltrisiloxane-N-glucoheptonamne (Si(3)N-GHA)] displayed higher extraction efficiency compared to other types of surfactants when it comes to being used in pretreatment methods. And a TSVUE method was established and optimised. In total, 131 constituents were identified in two BR herbs, of which 35 were unreported, and 11 were characterised as chemical markers. CONCLUSIONS: This method provides promising perspectives for rapidly identifying trace compounds in complex systems of traditional Chinese medicine (TCM), as well as for laying the foundation in the identification of similar herbs from the same species. Meanwhile, these findings serve as a promising application of trisiloxane surfactant vesicles in the extraction field of TCM.

A Gender Differentiated Analysis of Healthy Life Expectancy in South Asia: The Role of Greenhouse Gas Emission.

The sluggish progress concerning SDG-9 and SDG 13 has made South Asia an epicentre of household and ambient greenhouse gases emissions. Furthermore, the regional progress concerning attainment of SDG-3 is considerably low. The major research objectives are twofold. First, to explore the impact of GHGs emissions from agriculture, transportation, and manufacturing sector on disaggregated life expectancy. Second, to examine the mitigating impact of renewable energy use, trade integration, and human capital development for practice policy recommendations. These research objectives are realized by employing recently advanced cross-sectional auto regressive distributed lag (CS-ARDL) model on panel data of five South Asian countries such as Bangladesh, India, Pakistan, Nepal, and Sri Lanka from 1990 to 2019. The estimation outcome reveals that the emissions from transportation, manufacturing, and agricultural sectors significantly deteriorate healthy life expectancy of male and female healthy life expectancy in South Asia with different intensity. Especially, we find that long-run impact of GHG is more profound on male healthy life expectancy than female life expectancy. The result further shows that renewable energy and human capital substantially improve healthy life expectancy, whereas the effects of trade integration are insignificant. The finding of moderating variables shows that renewable energy, human capital development, and trade integration have high potential to reduce GHGs emissions. The findings of this study urge South Asia for investments in human capital development and renewable energy along with fostering regional integration to decrease GHG and improve healthy life expectancy.

Myopic loss aversion or equate-to-differentiate heuristic? A heuristic decision making model for both single and aggregated plays.

Myopic loss aversion (MLA)-a combination of myopic loss and a greater sensitivity to losses than gains-has been proposed to explain the equity premium puzzle and then extended to myopic prospect theory (MPT). However, such an expected-value/utility-based theory has been challenged and the underlying mechanism remains debatable. In the current study, we applied the modified equate-to-differentiate model to address this phenomenon. In Experiment 1, we first directly explored the relationship between individuals' degree of loss aversion and their investment amounts in risky lotteries for both single and repeated plays. We found no correlations between these variables; this was inconsistent with the MLA/MPT prediction. Experiment 2 showed that individuals' evaluation scores of the differences within the dimension (probability or outcome) that has larger differences highly predicted their investment behavior, which supported the equate-to-differentiate model.

Integrative analyses of widely targeted metabolomic profiling and derivatization-based LC-MS/MS reveals metabolic changes of Zingiberis Rhizoma and its processed products.

Zingiberis Rhizoma (ZR) has nutritional value and application potentiality, while Zingiberis Rhizoma Praeparatum (ZRP) and Carbonised Ginger (CG) are two main processed products of ZR based on different methods. Here, we performed a widely targeted metabolomics method with Sequential Windowed Acquisition of all Theoretical fragment ions (SWATH) mode to analyze differential metabolites in ZR, ZRP and CG. Additionally, the chemical derivatization was applied to characterize different submetabolomes and improve the separation effect and MS response of metabolites. In total, 369 metabolites were identified and divided into 14 categories, 104 of which were differential metabolites. Our results suggest that carbohydrates, nucleotides, organic acids, vitamins, lipids, indoles, alkaloids, and terpenes contributed to a downward trend after processing, but the maximum content of flavanones, phenylpropanes and polyphenols appeared in ZRP, and that of alcohols appeared in CG. These findings serve as promising perspectives for developing functional food in ZR, ZRP and CG.

Spinal dI4 Interneuron Differentiation From Human Pluripotent Stem Cells.

Spinal interneurons (INs) form intricate local networks in the spinal cord and regulate not only the ascending and descending nerve transduction but also the central pattern generator function. They are therefore potential therapeutic targets in spinal cord injury and diseases. In this study, we devised a reproducible protocol to differentiate human pluripotent stem cells (hPSCs) from enriched spinal dI4 inhibitory GABAergic INs. The protocol is designed based on developmental principles and optimized by using small molecules to maximize its reproducibility. The protocol comprises induction of neuroepithelia, patterning of neuroepithelia to dorsal spinal progenitors, expansion of the progenitors in suspension, and finally differentiation into mature neurons. In particular, we employed both morphogen activators and inhibitors to restrict or "squeeze" the progenitor fate during the stage of neural patterning. We use retinoic acid (RA) which ventralizes cells up to the mid-dorsal region, with cyclopamine (CYC), an SHH inhibitor, to antagonize the ventralization effect of RA, yielding highly enriched dI4 progenitors (90% Ptf1a+, 90.7% Ascl1+). The ability to generate enriched spinal dI4 GABAergicINs will likely facilitate the study of human spinal IN development and regenerative therapies for traumatic injuries and diseases of the spinal cord.

Quality control of Zingiberis Rhizoma and its processed products by UHPLC-Q-TOF/MS-based non-targeted metabonomics combining with SIBDV method.

Zingiberis Rhizoma (ZR) is a homologous plant with pungent tastes and aromas, which has unique nutritional value and tremendous application potentiality. Zingiberis Rhizoma Praeparatum (ZRP) and Carbonised Ginger (CG) are processed products of ZR through different processing methods, and they are commonly used ingredients in food supplements. This study used ZR, ZRP and CG from different batches to further understand composition differences after processing. Additionally, we performed non-targeted metabolomics-based profiling of gingerols by ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) in combination with multivariate analysis and compounds identification. In which, we developed a comprehensive SWATH-IDA bi-directionally verified (SIBDV) method integrating the advantages of Sequential Windowed Acquisition of all Theoretical fragment ions (SWATHTM) and traditional information-dependent acquisition (IDA) mode for characterization of gingerols. Potential chemical markers were selected by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) of chemometrics methods. After that, the threshold variable importance in projection (VIP) value and P value were employed to screen the valuable MS features for discriminating ZR, ZRP and CG. In total, 59 gingerols in the different samples were structurally identified. Results allowed the selection of 33 gingerols, which are nominated as novel markers for materials authentication in ZR, ZRP and CG. The analysis of the study showed that the content of gingerols showed a downward trend after processing, but shogaols and gingerone compounds had an upward trend, resulting in differences in application and pharmacodynamic efficacy. These findings provide promising perspectives in the quality control of ZR, ZRP and CG, as well as for laying the foundation in food design and development.

Special Application of ms/ms Method on Determination of Regiochemistry of Sugammadex Series Compounds: Synthesis, Isolation and Structural Characterization for Two Potential Oxidative Impurities in Sugammadex Sodium Finished Pharmaceutical Product.

Two oxidative degradation impurities of sugammadex sodium have been successfully synthesized under stress conditions and isolated by preparative high performance liquid chromatography, which would be extremely difficult to prepare stochiometrically by conventional methods due to their structural complexity. Characteristic fragmentation pattern observed by mass spectrometry for sugammadex series compounds helped distinguish the two regioisomeric di-sulfoxide impurities. Confirmed by nuclear magnetic resonance analysis, Impurity I was identified as ortho-disulfoxide sugammadex and Impurity II as meta-disulfoxide sugammadex. It is the first time detailed structures of these two impurities are reported. Additionally, HPLC analysis also indicated the observance of these two impurities in long-term stored sugammadex sodium finished pharmaceutical product but absence in three pilot batches of sugammadex sodium drug substance which met ICH requirements. The compounded analysis technique has proven to be successful and reliable, and we hope that it could be well applied to structure identification for other sugammadex impurities and will be beneficial for other researchers focusing on this field.

Development and Validation of a Radiomics Nomogram for Differentiating Pulmonary Cryptococcosis and Lung Adenocarcinoma in Solitary Pulmonary Solid Nodule.

OBJECTIVE: To establish a CT-based radiomics nomogram model for classifying pulmonary cryptococcosis (PC) and lung adenocarcinoma (LAC) in patients with a solitary pulmonary solid nodule (SPSN) and assess its differentiation ability. MATERIALS AND METHODS: A total of 213 patients with PC and 213 cases of LAC (matched based on age and gender) were recruited into this retrospective research with their clinical characteristics and radiological features. High-dimensional radiomics features were acquired from each mask delineated by radiologists manually. We adopted the max-relevance and min-redundancy (mRMR) approach to filter the redundant features and retained the relevant features at first. Then, we used the least absolute shrinkage and operator (LASSO) algorithms as an analysis tool to calculate the coefficients of features and remove the low-weight features. After multivariable logistic regression analysis, a radiomics nomogram model was constructed with clinical characteristics, radiological signs, and radiomics score. We calculated the performance assessment parameters, such as sensitivity, specificity, accuracy, negative predictive value (NPV), and positive predictive value (PPV), in various models. The receiver operating characteristic (ROC) curve analysis and the decision curve analysis (DCA) were drawn to visualize the diagnostic ability and the clinical benefit. RESULTS: We extracted 1,130 radiomics features from each CT image. The 24 most significant radiomics features in distinguishing PC and LAC were retained, and the radiomics signature was constructed through a three-step feature selection process. Three factors-maximum diameter, lobulation, and pleural retraction-were still statistically significant in multivariate analysis and incorporated into a combined model with radiomics signature to develop the predictive nomogram, which showed excellent classification ability. The area under curve (AUC) yielded 0.91 (sensitivity, 80%; specificity, 83%; accuracy, 82%; NPV, 80%; PPV, 83%) and 0.89 (sensitivity, 81%; specificity, 83%; accuracy, 82%; NPV, 81%; PPV, 82%) in training and test cohorts, respectively. The net reclassification indexes (NRIs) were greater than zero (p < 0.05). The Delong test showed a significant difference (p < 0.0001) between the AUCs from the clinical model and the nomogram. CONCLUSIONS: The radiomics technology can preoperatively differentiate PC and lung adenocarcinoma. The nomogram-integrated CT findings and radiomics feature can provide more clinical benefits in solitary pulmonary solid nodule diagnosis.