Analysis of Chemical Changes during Maturation of Amomum tsao-ko Based on GC-MS, FT-NIR, and FT-MIR.

Amomum tsao-ko Crevost et Lemaire (A. tsao-ko) is widely grown for its high nutritional and economic value. However, the lack of a scientific harvesting and quality control system has resulted in an uneven product quality. The present study was based on A. tsao-ko from four maturity stages from the same growing area, and its chemical trends and quality were evaluated using a combination of agronomic trait analysis, spectroscopy, chromatography, chemometrics, and network pharmacology. The results showed that A. tsao-ko was phenotypically dominant in October. Spectroscopy showed that the absorbance intensity at different maturity stages showed a trend of October > September > August > July. Further chemical differences between A. tsao-ko at different stages of maturity were found by chromatography to originate mainly from alcohol, aromatic, acids, esters, hydrocarbons, ketone, heterocyclic, and aldehydes. The network pharmacology results showed that the active ingredient for the treatment of obesity was present in A. tsao-ko and had high levels in A. tsao-ko in September and October. The results of this study provide a new idea for the comprehensive evaluation of A. tsao-ko and a theoretical basis for the harvesting and resource utilization of A. tsao-ko.

A rapid method for identification of Lanxangia tsaoko origin and fruit shape: FT-NIR combined with chemometrics and image recognition.

Lanxangia tsaoko's accurate classifications of different origins and fruit shapes are significant for research in L. tsaoko difference between origin and species as well as for variety breeding, cultivation, and market management. In this work, Fourier transform-near infrared (FT-NIR) spectroscopy was transformed into two-dimensional and three-dimensional correlation spectroscopies to further investigate the spectral characteristics of L. tsaoko. Before building the classification model, the raw FT-NIR spectra were preprocessed using multiplicative scatter correction and second derivative, whereas principal component analysis, successive projections algorithm, and competitive adaptive reweighted sampling were used for spectral feature variable extraction. Then combined with partial least squares-discriminant analysis (PLS-DA), support vector machine (SVM), decision tree, and residual network (ResNet) models for origin and fruit shape discriminated in L. tsaoko. The PLS-DA and SVM models can achieve 100% classification in origin classification, but what is difficult to avoid is the complex process of model optimization. The ResNet image recognition model classifies the origin and shape of L. tsaoko with 100% accuracy, and without the need for complex preprocessing and feature extraction, the model facilitates the realization of fast, accurate, and efficient identification.

Assessment of the relationship between plasma fibrinogen-to-albumin ratio and slow coronary flow phenomenon in patients without obstructive coronary artery disease.

BACKGROUND: Prior studies have suggested that the chronic inflammatory response has an important role in the pathophysiology of slow coronary flow phenomenon (SCFP). However, data are scarce regarding the role of plasma fibrinogen-to-albumin ratio (PFAR) in patients having SCFP without obstructive coronary artery disease (CAD). In this study, we investigated the relationship between PFAR and the presence of SCFP in patients without obstructive CAD. METHODS: From January 2021 to January 2023, we consecutively recruited 1085 patients without obstructive CAD according to the diagnostic and exclusion criteria. In total, SCFP was diagnosed in 70 patients. A 1:2 age-matched case-control study was then conducted using comparators without SCFP. Ultimately, this study enrolled 70 patients with angiographically normal coronary arteries and SCFP, along with 140 comparators with angiographically normal coronary arteries and normal coronary flow. Plasma fibrinogen and albumin levels were measured, and the PFAR was then calculated for each patient. RESULTS: PFARs were significantly greater in the SCFP group than in the comparators with normal coronary flow (82.8 ± 15.4 vs 73.1 ± 19.5, p < 0.001). PFAR increased with increasing numbers of vessels affected by SCFP. Multivariate logistic regression analysis showed that PFAR was an independent predictor of SCFP (odds ratio: 1.818, p = 0.015). Receiver operating characteristic (ROC) curve analysis indicated that PFAR showed a better predictive value of SCFP than fibrinogen or albumin, although not significantly (p > 0.05). CONCLUSION: PFAR is an independent predictor of SCFP in patients without obstructive CAD. PAFR could improve the predictive value of SFCP than albumin or fibrinogen alone, but not significantly.

An integrated chemical characterization based on FT-NIR, and GC-MS for the comparative metabolite profiling of 3 species of the genus Amomum.

BACKGROUND: The fruits and seeds of genus Amomum are well-known as medicinal plants and edible spices, and are used in countries such as China, India and Vietnam to treat malaria, gastrointestinal disorders and indigestion. The morphological differences between different species are relatively small, and technical characterization and identification techniques are needed. RESULTS: Fourier transform near infrared spectroscopy (FT-NIR) and gas chromatography-mass spectrometry (GC-MS), combined with principal component analysis and two-dimensional correlation analysis were used to characterize the chemical differences of Amomum tsao-ko, Amomum koenigii, and Amomum paratsaoko. The targets and pathways for the treatment of diabetes mellitus in three species were predicted using network pharmacology and screened for the corresponding pharmacodynamic components as potential quality markers. The results of "component-target-pathway" network showed that (+)-Nerolidol, 2-Nonanol, α-Terpineol, α-Pinene, 2-Nonanone had high degree values and may be the main active components. Partial least squares-discriminant analysis (PLS-DA) was further used to select for differential metabolites and was identified as a potential quality marker, 11 in total. PLS-DA and residual network (ResNet) classification models were developed for the identification of 3 species of the genus Amomum, ResNet model is more suitable for the identification study of large volume samples. SIGNIFICANCE: This study characterizes the differences between the three species in a visual way and also provides a reliable technique for their identification, while demonstrating the ability of FT-NIR spectroscopy for fast, easy and accurate species identification. The results of this study lay the foundation for quality evaluation studies of genus Amomum and provide new ideas for the development of new drugs for the treatment of diabetes mellitus.

Effects of Different Pre-drying and Drying Methods on Volatile Compounds in the Pericarp and Kernel of Amomum tsao-ko.

The effects of twelve different pre-drying and drying methods on the chemical composition in the pericarp and kernel of Amomum tsao-ko were studied. The volatile components were isolated from the samples by simultaneous distillation and extraction and analyzed by gas chromatography-mass spectrometry (GC-MS). Sixty and thirty-eight compounds were identified from pericarp and kernel, respectively, and the main constituents were oxygenated monoterpenes. These compounds were not only significantly affected by pre-drying and drying methods but also varied in content due to different tissue locations. The total volatile content of pericarp varied from 0.70 to 1.55%, with the highest obtained by microwave-dried samples (150 W) and the lowest in freeze-dried samples. The total volatile content of the kernel varied from 6.11 to 10.69%, with the highest content obtained during sun drying (SD) and the lowest content in samples treated with boiling water for 2 min. Oxygenated monoterpenes were the highest compounds in pericarp and kernel, which were also the most affected by drying methods. The highest content of oxygenated monoterpenes in the pericarp (0.77%) could be obtained by boiling water treatment for 5 min, and the highest content of oxygenated monoterpenes in the kernel (7.48%) could be obtained by SD. Additionally, the main components such as 1,8-cineole, 2-carene, (Z)-citral, nerolidol, (Z)-2-decenal, (E)-2-dodecenal, citral, (E)-2-octenal, 4-propylbenzaldehyde, and phthalan showed remarkable variations in pre-drying and drying methods.

The Role of Spinal GABAB Receptors in Cancer-Induced Bone Pain in Rats.

Cancer-induced bone pain (CIBP) remains a major challenge in advanced cancer patients because of our lack of understanding of its mechanisms. Previous studies have shown the vital role of γ-aminobutyric acid B receptors (GABABRs) in regulating nociception and various neuropathic pain models have shown diminished activity of GABABRs. However, the role of spinal GABABRs in CIBP remains largely unknown. In this study, we investigated the specific cellular mechanisms of GABABRs in the development and maintenance of CIBP in rats. Our behavioral results show that acute as well as chronic intrathecal treatment with baclofen, a GABABR agonist, significantly attenuated CIBP-induced mechanical allodynia and ambulatory pain. The expression levels of GABABRs were significantly decreased in a time-dependent manner and colocalized mostly with neurons and a minority with astrocytes and microglia. Chronic treatment with baclofen restored the expression of GABABRs and markedly inhibited the activation of cyclic adenosine monophosphate (cAMP)-dependent protein kinase and the cAMP-response element-binding protein signaling pathway. PERSPECTIVE: Our findings provide, to our knowledge, the first evidence that downregulation of GABABRs contribute to the development and maintenance of CIBP and restored diminished GABABRs attenuate CIBP-induced pain behaviors at least partially by inhibiting the protein kinase/cAMP-response element-binding protein signaling pathway. Therefore, spinal GABABR may become a potential therapeutic target for the management of CIBP.

[Study on the Genetic Relationship of Panax Notoginseng and Its Wild Relatives Based on Fourier Translation Infrared Spectroscopy].

Wild relatives play a very important role in enriching germplasm resources and improving the quality and yield of cultivated species. In this paper, the genetic relationship between Panax notoginseng and its wild relatives has been investigated by using Fourier transform infrared (FTIR) spectroscopy in order to provide theoretical bases in the variety improvement of P. notoginseng as well as the development and utilization of germplasm resources. The FTIR spectra of P. notoginseng and its wild relatives (P. japonicus var. major, P. stipuleanatus, P. vietnamensis, P. japonicus var. bipinnatifidus) as well as Panax notoginsenosides were collected. The original infrared spectra of P. notoginseng and its wild relatives were pretreated by automatic baseline correction, smoothing, ordinate normalization and second derivative. The genetic relationship between P. notoginseng and its wild relatives has been studied together with the aid of principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and hierarchical cluster analysis (HCA). By comparing the infrared spectra of P. notoginseng with that of panax notoginsenosides, some common peaks such as 3 400, 2 930, 1 635, 1 385, 1 075 and 927 cm-1 has been found. It showed that the peak heights of P. notoginseng samples may relate with the content of panax notoginsenosides. The original infrared spectra of P. notoginseng are similar to its wild relatives and the absorption peaks of the functional groups of C­H, CO, O­H, C­N and C­O were presented. There were some differences in the fingerprint region (1 800～500 cm-1) of the second derivative spectra of these five species samples. The characteristic absorption peaks such as 1 385 and 784 cm-1 has an obviously differentiation. Then the fingerprint region of second derivative spectra is subjected to be analyzed by PCA and PLS-DA. By comparing the 3D score plots of these two methods, the classification result of PLS-DA is significantly better than PCA. In addition, the result of HCA which based on the six principal components of PLS-DA has shown that P. japonicus var. major and P. vienamensis have close relationship with P. notoginseng while P. stipuleanatus and P. japonicus var. bipinnatifidus are far from P. notoginseng. The use of Fourier transform infrared spectroscopy combined with chemometrics methods could effectively investigate the genetic relationship between P. notoginseng and its wild relatives. Furthermore, it could provide reference for the research of medicinal plants.

Aurora-A identifies early recurrence and poor prognosis and promises a potential therapeutic target in triple negative breast cancer.

Triple negative breast cancer (TNBC) acquires an unfavorable prognosis, emerging as a major challenge for the treatment of breast cancer. In the present study, 122 TNBC patients were subjected to analysis of Aurora-A (Aur-A) expression and survival prognosis. We found that Aur-A high expression was positively associated with initial clinical stage (P = 0.025), the proliferation marker Ki-67 (P = 0.001), and the recurrence rate of TNBC patients (P<0.001). In TNBC patients with Aur-A high expression, the risk of distant recurrence peaked at the first 3 years and declined rapidly thereafter, whereas patients with Aur-A low expression showed a relatively constant risk of recurrence during the entire follow-up period. Univariate and multivariate analysis showed that overexpression of Aur-A predicted poor overall survival (P = 0.002) and progression-free survival (P = 0.012) in TNBC. Furthermore, overexpression of Aur-A, associated with high Ki-67, predicted an inferior prognosis compared with low expression of both Aur-A and Ki-67. Importantly, we further found that Aur-A was overexpressed in TNBC cells, and inhibition of this kinase inhibited cell proliferation and prevented cell migration in TNBC. Our findings demonstrated that Aur-A was a potential therapeutic target for TNBC and inhibition of Aur-A kinase was a promising regimen for TNBC cancer therapy.

[Ultraviolet absorption spectrum analysis and identification of medicinal plants of Paris].

Species of Paris are important medicinal plants of China. They possess anticancer, hot alexipharmic, detumescence, acesodyne, and arrest blood and remove blood stasis effects. They are the main raw material for several Chinese patent drugs such as "Yunnan Baiyao", "Gong Xue Ning", "Re Du Qing" and "Ji De Sheng Sheyaopian". The present paper, through optimizing the chloroform, absolute ethyl alcohol and water extraction condition of Paris by orthogonal test L3(4) (16), using mean value, smoothness and second differential methods on the observed UV spectrum, to inspects the RSD of stability and repeatability of different waveband. By SIMCA and the common and variant peak ratio dual index sequence analysis method, it evaluated the quality and quantity of Paris. The results showed that at the time of 50, 40 and 50 min, chloroform, absolute ethyl alcohol and water had the highest extraction ratios. Within 20 h, the RSDs of stability were 0.06-1.88, 0.05-2.42 and 0.03-0.35; the RSDs of accuracy were 0-1.48, 0.05-0.37 and 0.09-0.44; and the RSDs of repeatability were 0-1.23, 0.04-0.30 and 0.12-0.25 respectively. The qualitative analysis revealed large differences between different Paris species and different areas. The quantitative analysis indicated that the highest common peak ratio among the Paris samples was 80.00% and the lowest variant peak ratio was 6.25%. The method evaluated Paris of different species and from different producing areas, and also quantitatively assessed the arbitrary two samples, clarified the similarity between the species and areas of Paris, which provided basis of distinguishing the real and false, identification of variety and quality evaluation for Chinese herbal medicine.

[Species and ecological control of disease on cultivated Gentiana rigescens in Yunnan].

OBJECTIVE: To find out variety of the fungal diseases of cultivated Gentiana rigescens and provide important basis for prevention. METHODS: The diseases were diagnosed based on field investigate, symptoms observation, pathogen isolation, determination the size of morphological and verification following the Koch's Postulate procedures. RESULTS: Anthracnose (Colletotrichum gloeosporioides), grey mould (Botrytis cinerea), brown spot (Alternaria tenuis), rust (Aecidiumpers), circular spot (Pestalotiopsis), leaf blight (Stemphylium, Ascochyta, Pleospora) and nematodes (Heterodera spp., Meloidogyne spp.) were found on Gentiana rigescens. Anthracnose was the first main disease, the diseased plant rate was over 40% and disease severity was 4 - 5 degree and second disease was rust, incidence of rate was less 10% and other diseases rate was not enough 2%. CONCLUSION: All these diseases on Gentiana rigescens are reported for the first time and Gentiana rigescens is the new host plant of the diseases.

Low expression of Beclin 1, associated with high Bcl-xL, predicts a malignant phenotype and poor prognosis of gastric cancer.

Recent studies have suggested that dysregulation of autophagy plays a pivotal role in tumorigenesis. Here, we determined the prognostic value of autophagy-related protein Beclin 1 in gastric cancer. A total of 153 primary gastric cancer patients were subjected to analysis of Beclin 1 expression and survival prognosis. Among them, 68 patients were assigned randomly and used as a training set to generate a cutoff score for Beclin 1 expression by receive operating characteristic (ROC) curve analysis. The ROC-generated cutoff score was subjected to analyze the association of Beclin 1 with clinical characteristics and patient outcome. In a testing set (n = 85) and overall patients (n = 153), both univariate and multivariate analysis found that low expression of Beclin 1 predicted adverse overall survival and progression-free survival for gastric cancer patients. Furthermore, in each stage of gastric cancer patients, Beclin 1 expression was a prognostic indicator in patients with stage II, III and IV. Importantly, a reverse relationship between Beclin 1 and Bcl-xL expression was demonstrated. In patients of elevated Bcl-xL expression, a subset with lower Beclin 1 expression displayed an inferior overall survival and progression-free survival than those with higher Beclin 1 expression. Thus, our data demonstrated that low expression of Beclin 1, associated with high Bcl-xL, played as an independent biomarker, contributing to a more aggressive cancer cell phenotype and poor prognosis for gastric tumor.

Regulation of spinal neuroimmune responses by prolonged morphine treatment in a rat model of cancer induced bone pain.

Cancer induced bone pain (CIBP) is a major clinical problem. Although opioids remain the principal axis in drug therapies for CIBP, their sustained application is known to induce cellular and molecular adaptations including enhanced neuroimmune reactivity. This is generally characterized by glial activation and proinflammatory cytokine production which frequently results in pharmacological tolerance. This research was performed to investigate spinal neuroimmune responses after prolonged systemic morphine treatment in a rat model of CIBP. The model was established using a unilateral intra-tibia injection of Walker 256 mammary gland carcinoma cells. Subcutaneous morphine was repeatedly administered from postoperative days 14 to 19. Mechanical allodynia to von Frey filaments and ambulatory pain scores were recorded to investigate changes of nociceptive behaviors. Spinal glial activation was detected by immunohistochemistry and real-time PCR; the production of proinflammatory cytokines (IL-1beta and TNF-alpha) was examined through real-time PCR and ELISA. Results showed that chronic morphine use failed to elicit analgesic tolerance in the rat CIBP model. Moreover, the treatment had no significant influence on the activated spinal glia morphology, cell density and expression of special cytomembrane markers, whereas it significantly down-regulated the local proinflammatory cytokine production at the mRNA and protein level. Collectively, these data suggest that chronic morphine treatment in CIBP is not concomitant with pharmacological tolerance, at least partially because the treatment fails to amplify spinal neuroimmune responses.

Evaluation of side effects through selective ablation of the mu opioid receptor expressing descending nociceptive facilitatory neurons in the rostral ventromedial medulla with dermorphin-saporin.

Descending facilitation from the rostral ventromedial medulla (RVM) contributes to some pathological pain states. The intra-RVM microinjection with dermorphin-saporin could specifically abolish this facilitation in rodent models by selectively ablating the RVM neurons expressing mu opioid receptors. Thus, this targeted lesion may be an alternative mechanism-based approach for intractable pain. This research was performed to investigate potential side effects after a single intra-RVM application of dermorphin-saporin in rats. Results showed though some acute cardiovascular signs were observed with dermorphin-saporin, the treatment exhibited no long-lasting significant influence on some physiological functions for up to 3-month observation period, including normal sensory function, locomotor activity, ingestive behaviors, body weight, rectal temperature, respiratory rate, heart rate, systolic blood pressure, cardiac structure and function. Moreover, there were only mild microglial responses on day 7 post-microinjection, while no significant increase in the immunostaining of astrocytes and no noticeable up-regulation in the production of proinflammatory cytokines were detected in the RVM treated with dermorphin-saporin. Taken together, these data would suggest that this selective ablation of mu opioid receptor bearing descending facilitatory neurons in the RVM with dermorphin-saporin did not elicit the long-standing evident adverse toxicity in terms of some physiological parameters and neurochemical alterations we determined, plausibly providing us a safe and reliable approach to treat some intractable pain.

[Construction of rat constitutively active recombinant caspase-3 and investigation of its pro-apoptotic effect in vitro].

OBJECTIVE: To investigate the apoptosis induction effect by the rat recombinant caspase-3. METHODS: Reversed rat caspase-3 gene was gained by settling the small subunit prior to the large one through recombinant PCR, and cloned into the expression vector to transfect human 293T cells and rat immortalized neural progenitor cells. The expression and pro-apoptotic effect of recombinant caspase-3 was observed by the changes of morphology of the transfected cells through immunofluorescence and analyzed by Annexin V-FITC staining, Flowcytometry and MTT assay. RESULTS: The transfected cells presented obvious apoptosis as detected by immunofluorescence. MTT assay showed that the proliferation of recombinant caspase-3 transfected cells was significantly inhibited [(48.35 +/- 0.16)%, (44.61 +/- 0.15)%] (P < 0.05). Annexin V-FITC staining revealed that the percentage of apoptotic cells in the transfectants of recombinant caspase-3 gene was [(30.7 +/- 1.5)%, (16.0 +/- 1.0)%] (P < 0.05), which was much higher than that of control cells. CONCLUSIONS: Rat recombinant caspase-3 can be expressed and induce apoptosis effectively, and used safely both in vitro and in vivo. It can also cause neural cells to death.