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Aim To investigate whether alisol A (AA) could improve the blood brain barrier (BBB) mediated cortex cerebral ischemia-repeifusion injury (CIRI) by inhibiting matrix metalloproteinase 9 (MMP-9). Methods The global cerebral ischemia- reperfusion (GCI/R) model in mice was established, and the AA was intragastric injected subsequently for seven days. The modified neurological severity scores (mNSS), open field test and Y-maze test were applied to detect neurological function. Magnetic resonance spectroscopy (MRS) was used to detect relevant neu- rosubstance metabolism in cortex of mice. Transmission electron microscope (TEM) was employed to observe the ultrastructure of BBB in cortex. Western blot and immunohistochemistry were used to detect the MMP-9 level in cortex. The binding possibility of A A and MMP-9 was determined by molecular docking. Results Compared with Sham group, mice in GCI/R group have an increased mNSS score but decreased at total distance and center distance to total distance ratio in open field test as well as alternation rate in Y-maze test (P<0.01). While mice in GCI/R + AA group have a decreased mNSS score but increased at total distance and center distance to total distance ratio in open field test as well as alternation rate in Y-maze test (P<0.01) compared with GCI/R group. MRS results found that in cortex of GCI/R group mice, the level of GABA and NAA significantly decreased while the Cho, mI and Tau level increased (P<0.01). Whereas in GCI/R + AA group mice, the GABA and NAA level increased and the Cho, ml and Tau decreased significantly (P<0.01). By TEM we observed that the basilemma of cerebral microvessels collapsed, the lumen narrowed, the endothelial cells were active and plasma membranes ruffled, gaps between cells were enlarged and tight junctions were damaged and the end feet of astrocytes were swollen in GCI/R group mice. While in GCI/R + AA group mice, the lumen was filled, plasma membranes of endothelial cells were smooth, tight junctions were complete and end feet of astrocytes were in normal condition. Western blot and immunohistochemistry both found that the MMP-9 level increased in GCI/R group mice (P < 0.01) and decreased in GCI/R + AA group mice (P < 0.05). Molecular docking proved the binding between aliso A and MMP9 through TYR-50 and ARG-106, and the binding energy was calculated as -6.24 kcal · mol
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Purpose@#Deceased donor liver transplantation (DDLT) recipients in Korea are generally sicker due to an increasing organ shortage. In the present study, the risk factors for early 30-day liver graft failure after DDLT were identified. @*Methods@#From August 2017 to February 2021, 265 adult DDLTs were performed. The characteristics of patients with and without 30-day graft failure were compared. @*Results@#Liver graft failure occurred in 11 patients (17.7%) after DDLT. Baseline and perioperative characteristics of donors and recipients were not statistically significantly different between the 2 groups. The cumulative graft and overall survival rates at 6 months were 83.9% and 88.7%, respectively. Multivariate analysis showed ventilator support in the pretransplant period was a predisposing factor for 30-day graft failure after DDLT. @*Conclusion@#Present study indicates that cautious decision is required when allocating DDLT in critically ill patients on mechanical ventilatory support.
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OBJECTIVE@#To explore the chronic injury and its possible mechanism of ionizing radiation on multipotent hematopoietic progenitor cells (MPPs) by determining the related indicators of MPPs in bone marrow of mice post-radiation.@*METHODS@#Sixteen C57BL/6 adult mice were randomly divided into normal control and irradiation groups, 8 mice in each group. The mice in irradiation group were exposed to 6 Gy X-ray. The proportion of bone marrow MPPs, their apoptosis and proliferation 2 months after irradiation were detected by flow cytometry. Mitochondrial activity and levels of reactive oxygen species (ROS) in each MPPs population were detected by Mitotracker Red and DCFDA probes, and the senescent state of MPPs in the bone marrow was analyzed.@*RESULTS@#Ionizing radiation could reduce the proportion of MPPs in mouse bone marrow. The proportions and numbers of MPP1, MPP3 and MPP4 in the bone marrow were significantly decreased after whole-body irradiation with 6 Gy X-ray (P<0.05). In addition, radiation significantly reduced the colony-forming capacity of MPPs in bone marrow (P<0.05), the proportions of apoptotic cells in the MPP1 and MPP4 cell populations increased significantly in the bone marrow (P<0.05). The activity of mitochondria was significantly reduced in the bone marrow MPP2, MPP3 and MPP4 cell populations compared with that of the control group (P<0.05). It was also found that the radiation could significantly increase the ROS levels of MPPs in bone marrow, and the content of ROS in the MPP2, MPP3 and MPP4 cell population of the bone marrow was significantly increased(P<0.05). The senescent cells ratios of MPP1, MPP3 and MPP4 cells in the bone marrow after irradiation were significantly higher than those in the control group (P<0.05).@*CONCLUSION@#Ionizing radiation can cause chronic MPPs damage in mice, which is closely associated with persistent oxidative stress, cells apoptosis, and cellular senescence.
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Mice , Animals , Bone Marrow , Reactive Oxygen Species , Mice, Inbred C57BL , Hematopoietic Stem Cells , Whole-Body Irradiation , Radiation, Ionizing , Bone Marrow CellsABSTRACT
ObjectiveTo reveal the mechanism of action of Huangqi Guizhi Wuwutang in the treatment of rheumatoid arthritis by pharmacological research based on its clinical application. MethodThe collagen-induced arthritis (CIA) rat model was established by injecting bovine type Ⅱ collagen and Freund's adjuvant at the tail, and was treated with different concentrations of Huangqi Guizhi Wuwutang. The rats were randomly divided into blank group, model group, methotrexate (0.9 mg·kg-1) group, and Huangqi Guizhi Wuwutang low- and high-dose (5.13, 20.52 g·kg-1·d-1) groups, with continuous intragastric administration for 4 weeks. The degree of joint swelling, weight, degree of foot swelling and arthritis index score were determined and the pathological changes of ankle joints were detected by hematoxylin and eosin (HE) staining to observe the therapeutic effect of Huangqi Guizhi Wuwutang on rheumatoid arthritis. In addition, enzyme-linked immunosorbent assay (ELISA) and Western blot were used to measure the expression of interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 10 (IL-10) and tumor necrosis factor-α (TNF-α) in serum and the expression of nuclear factor kappa-B (NF-κB) pathway related proteins in synovial tissue, respectively to clarify the molecular mechanism of Huangqi Guizhi Wuwutang in the treatment of rheumatoid arthritis. ResultCompared with the conditions in blank group, the body weight and IL-10 level were decreased (P<0.01), and the degree of foot swelling and arthritis index score, the levels of IL-1β, IL-6 and TNF-α, and the expression of NF-κB pathway related proteins were increased (P<0.01,) in the model group, with impaired morphology and function of the ankle joint. Additionally, compared with the model group, Huangqi Guizhi Wuwutang low- and high-dose groups had increased body weight of rats and IL-10 level (P<0.01), and reduced degree of foot swelling and arthritis index score (P<0.05, P<0.01), levels of IL-1β, IL-6 and TNF-α (P<0.01) and expression of NF-κB pathway related proteins (P<0.05, P<0.01), with improved function and morphology of the ankle joint. ConclusionHuangqi Guizhi Wuwutang can significantly alleviate joint inflammatory injury by down-regulating NF-κB pathway and reducing the inflammatory response in CIA rats.
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ObjectiveTo observe the inhibitory effect of modified Xiao Xianxiongtang on epithelial-mesenchymal transition (EMT) of human gastric cancer MGC803 cells and its relationship with secretory glycoprotein Wnt/β-catenin pathway. MethodThe BALB/c nude mice were implanted with human gastric cancer MGC803 cell suspension in the heterotopic subcutaneous position for inducing tumor. After successful modeling, they were randomly divided into the model group, low-, medium-, and high-dose (16.0,32.0,and 64.0 g·kg-1) groups of modified Xiao Xianxiongtang, and capecitabine (400 mg·kg-1) group, with eight mice in each group, and gavaged with the corresponding drugs, once per day, for 28 consecutive days. Those in the capecitabine group received one-week discontinuation after every two weeks of treatment. The general state and body weight of the nude mice were observed, and the transplanted tumor volume was measured. After being killed, they were weighed and the tumor inhibition rate was calculated. Hematoxylin-eosin (HE) staining was carried out for observing the pathological changes in transplanted tumor tissues. The gene and protein expression levels of Wnt1 and β-catenin were detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot, followed by the determination of matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), N-cadherin, E-cadherin, Vimentin, and Snail protein expression by Western blot. The expression levels of cyclooxygenase 2 (COX2) and prostaglandin E2 (PGE2) were detected by enzyme-linked immunosorbent assay (ELISA). ResultIt was found that the transplanted tumor in each group showed different growth trends with time, with the most obvious growth observed in the model group. Compared with the model group, the low-, medium-, and high-dose modified Xiao Xianxiongtang groups exhibited reduced tumor volume and slowed growth to varying degrees over time. After medication for days 7,14,21,and 28, the tumor volumes in the low- and high-dose modified Xiao Xianxiongtang groups and capecitabine group declined (P<0.05, P<0.01), and that in the medium-dose Xiao Xianxiongtang group was also remarkably reduced after medication for days 14,21,and 28 (P<0.01). Compared with the model group, the high-dose modified Xiao Xianxiongtang group and capecitabine group showed a significant reduction in the relative tumor volume after treatment for days 7,14,21,28 (P<0.01), and the low- and medium-dose modified Xiao Xianxiongtang groups also presented with decreased relative tumor volume after treatment for days 14,21,28 (P<0.05, P<0.01). Compared with the model group, the modified Xiao Xianxiongtang at low, medium, and high doses and capecitabine all increased the tumor inhibition rate to varying degrees (P<0.01), down-regulated the mRNA and protein expression levels of Wnt1 and β-catenin in tumor tissue (P<0.05, P<0.01) and protein expression levels of MMP-9, VEGF, N-cadherin, Vimentin, and Snail (P<0.05, P<0.01), up-regulated E-cadherin protein expression (P<0.05, P<0.01), and reduced COX2 and PGE2 contents (P<0.05, P<0.01). ConclusionModified Xiao Xianxiongtang inhibits the EMT of human gastric cancer MGC803 cell-transplanted tumor, which may be related to Wnt/β-catenin pathway.
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ObjectiveTo explore the effect of Xiao Xianxiongtang (XXXT) on the transforming growth factor (TGF)-β1-induced invasion, metastasis, and epithelial-mesenchymal transition (EMT) of gastric cancer MGC-803 cells and the underlying mechanism. MethodThe molecular docking between XXXT and nuclear factor of activated T cells (NFAT) was performed by CB-DOCK (http://clab.labshare.cn/cb-dock/). The invasion and metastasis model of MGC-803 cells was established with 10 μg·L-1 TGF-β1. MGC-803 cells were classified into blank group, model group, 0.1 g·L-1 XXXT group, 0.2 g·L-1 XXXT group, and 0.4 g·L-1 XXXT group. For further clarifying the key role of Wnt5a/Ca2+/NFAT signaling pathway in the inhibition of XXXT on gastric cancer, MGC-803 cells were transfected with Wnt5a overexpression plasmid, and then the cells were classified into blank plasmid group, Wnt5a-OE group, blank plasmid + XXXT (0.4 g·L-1) group, and Wnt5a-OE + XXXT (0.4 g·L-1) group. Cell viability was determined by cell counting kit-8 (CCK-8) assay, cell invasion and migration ability by Transwell invasion assay and wound healing assay, expression of EMT-related proteins (E-cadherin, N-cadherin, Vimentin, Snail) and Wnt5a/Ca2+/NFAT signaling pathway-related key proteins [Wnt5a, calcineurin (CaN), NFAT1, and p-NFAT1] by Western blot, and changes in intracellular Ca2+ concentration by immunofluorescence assay. ResultMolecular docking suggested that XXXT acted on Wnt5a/Ca2+/NFAT signaling pathway. XXXT (0.1, 0.2, 0.4 g·L-1) significantly promoted the loss of MGC-803 cell viability (P<0.05,P<0.01). It inhibited cells from invading the transwell lower chamber and slowed down the healing of cell wounds in a dose-dependent manner (P<0.05, P<0.01). Moreover, it promoted the expression of E-cadherin while suppressed the expression of N-cadherin, Vimentin, and Snail (P<0.05, P<0.01). Further experiments showed that XXXT could inhibit the expression of Wnt5a, CaN, NFAT1, and p-NFAT1, and reduce the nuclear expression of NFAT1 and the transcription activity mediated by NFAT1, so as to reduce the cellular Ca2+ concentration (P<0.05, P<0.01). XXXT can reverse the effect of Wnt5a (P<0.05, P<0.01). ConclusionXXXT can attenuate the invasion, metastasis, and EMT of MGC-803 cells via the Wnt5a/Ca2+/NFAT pathway, thereby weakening the tumor-promoting effect of TGF-β1. In summary, XXXT may exert therapeutic effect on gastric cancer by regulating the invasion, metastasis, and EMT of gastric cancer cells.
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Objective:This studu aims to investigate the effect of aqueous extract of modified Xiao Xianxiongtang on the epithelial mesenchymal transition(EMT) and the change of its invasion and migration ability of human gastric cancer MGC-803 cells mediated by transforming growth factor-<italic>β</italic><sub>1</sub>(TGF-<italic>β</italic><sub>1</sub>),and to explore the mechanism of regulating Wnt5a/Ca<sup>2+</sup>/ activated T-cell nuclear factor(NFAT) signaling pathway to inhibit EMT and invasion and metastasis of MGC-803 cells. Method:TGF-<italic>β</italic><sub>1</sub>(10 μg·L<sup>-1</sup>)was used to induce EMT and the invasion and metastasis model of human gastric cancer MGC-803 cells. Transwell chamber experiment, scratchhealing experiment, Western blot and immunofluorescence assay were used to detect cell invasion and migration ability, expression of EMT marker protein and key protein expression of Wnt5a/Ca<sup>2+</sup>/NFAT signaling pathway, and intracellular Ca<sup>2+</sup> concentration. Result:Compared with the blank group, TGF-<italic>β</italic><sub>1</sub> could significantly enhance the invasion and migration ability of MGC-803 cells(<italic>P</italic><0.01), down-regulate the level of E-cadherin(<italic>P</italic><0.01), up-regulate protein expressions of N-cadherin, Snail and Vimentin(<italic>P</italic><0.01), and induce cell Wnt5a, calcineurin (CaN), total protein of activated T-cell nuclear factor 1(NFAT1), up-regulation of phosphorylated proteins p-NFAT1 and NFAT1 nucleoprotein and intracellular accumulation of Ca<sup>2+</sup>(<italic>P</italic><0.01). Compared with the TGF-<italic>β</italic><sub>1</sub> group, modified Xiao Xianxiongtang (10, 20, 40 mg·L<sup>-1</sup>) could significantly inhibit this phenomenon,and 40 mg·L<sup>-1</sup> had the best effect(<italic>P</italic><0.05,<italic>P</italic><0.01).The specific inhibitors of Wnt5a/Ca<sup>2+</sup>/NFAT signaling pathway (<italic>R</italic>)-(+)-Bay-K-8644 and modified Xiao Xianxiongtang (40 mg·L<sup>-1</sup>) could significantly inhibit theinvasion and migration of MGC-803 cells mediated by TGF-<italic>β</italic><sub>1</sub>, up-regulate the level of E-cadherin, and down-regulate expressions of N-cadherin, Snail, Vimentin, Wnt5a, CaN and NFAT1 proteins and reduce the intracellular accumulation of Ca<sup>2+</sup>(<italic>P</italic><0.05,<italic>P</italic><0.01).Moreover, (<italic>R</italic>)-(+)-Bay-K-8644 combined with modified Xiao Xianxiongtang (40 mg·L<sup>-1</sup>) had stronger inhibitory effect(<italic>P</italic><0.05,<italic>P</italic><0.01). Conclusion:These results suggest that modified Xiao Xianxiongtang can inhibit the EMT mediated by TGF-<italic>β</italic><sub>1</sub> via Wnt5a/Ca<sup>2+</sup>/NFAT signaling pathway,thereby reducing the invasion and migration ability of MGC-803 cells.
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Objective@#This study aims to investigate the epidemiological distribution of HFMD and quantify the association of temperature with the incidence of children’s HFMD in Nanjing, China.@*Methods@#Daily counts of HFMD in children under 5 years and daily meteorological variables during 2011-2016 were obtained. Descriptive statistics were used to describe the epidemiological characteristics and distributed lag non linear model (DLNM) was used to assess the associations of temperature on HFMD cases.@*Results@#A total of 104 977 HFMD cases aged 0-5 years were reported in Nanjing during the study period and the male to female sex ratio was 1.49∶1. The average annual incidence was 213.5 per 100 000. A bimodal seasonal pattern was observed and the south and west were found to be the high incidence areas in the city. Of these laboratory confirmed enteroviruses positive cases, 32.5% cases were positive for EV-A71 infections, 29.1% cases were positive for CV-A16 infections and 38.4% cases were positive of other enteroviruses infections. The temperature HFMD relationships were non linear and showed obvious lag effects. The cumulative relative risk presented as an approximately inverted U shape over 14 days and peaked at 25.7 ℃ with value of 2.71(95%CI=1.93-3.81). Subgroup analyses revealed that males and children aged <1 year were more vulnerable to temperature variations.@*Conclusion@#Epidemiological characteristics of HFMD among children aged 0-5 years old in Nanjing presented temporal and regional distribution. The temperature has significant impact on children’s HFMD occurrence.
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The paper aims to establish the method to determine the monosaccharide composition and monosaccharide ratio in propylene glycol alginate sodium sulphate (PSS). Samples were hydrolyzed with trifluoroacetic acid, neutralized with sodium hydroxide solution after the reaction conditions for sample pretreatment were optimized via orthogonal analysis. A high performance anion exchange chromatograghy (HPAEC) coupled with pulsed amperometric detector (PAD) was performed on a CarboPac®PA20, using 200 mmol·L-1 sodium hydroxide solution and 1 mol·L-1 sodium acetate solution as mobile phase. The established HPAEC-PAD method was validated by testing the linear relationship, precision and accuracy, and showed exclusive, sensitive, rapid and wide use. The monosaccharide composition of PSS from different manufacture can be accurately determined with great significance for the structural identification of PSS.
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Deamidation is one of the most common degradation impurities in protein and peptide drugs. The deamidation of glutamine and asparagine in the protein sequence can lead to changes in the chemical and biological properties of the protein. However, the rutine trypsin-based pretreatment process can significantly increase the artificial deamidation impurities during the digestion process, resulting in high determination level. In this study, after optimizing the conditions of Glu-C enzymatic hydrolysis, we obtained the best enzymatic conditions under acidic condition and the artificial deamidation impurities significantly reduced in digestion process, identified the deamidation site (N48). Through the methodological investigation and comparison of the measurement results of different methods, the specificity, reproducibility and accuracy of the method are verified. The method established in this research has laid a solid foundation for the accurate determination of deamidation impurities in cobratide and its similar protein peptide biochemical drugs.
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Despite the concept of membrane anatomy has been widely used in minimally invasive colorectal surgery, the definition of membrane anatomy and the establishment of membrane plane remain controversial. Therefore, it is difficult to establish a unified theoretical system of membrane anatomy. Through embryological studies and anatomical findings on the integrity and continuity of membranes, we try to discuss the theoretical system of membrane anatomy in colorectal surgery from three aspects: membrane anatomical system, membrane anatomical elements and membrane anatomical mechanism. The establishment of a unified theoretical system of membrane anatomy will not only contribute to the standardization operative procedures, but also to the establishment of uniform surgical standards for colorectal cancer.
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Humans , Colorectal Surgery , Digestive System Surgical Procedures , Fascia , Mesentery , Minimally Invasive Surgical ProceduresABSTRACT
Objective To investigate the relationship between human papillomavirus (HPV) infection and cervical lesions, and to explore the application value of HPV typing in cervical disease screening. Methods Colposcopic examination and cervical biopsy were performed in 1 851 cases of HPV positive patients from January 2017 to September 2018 in the cervical disease clinic of Shanghai First People's Hospital.Pathological diagnosis was used as the gold standard to analyze the distribution of HPV subtypes and the relationship between HPV subtypes and cervical lesions. Results Among 1 851 patients, high-risk human papillomavirus (HPV) infection was the main type.The detection rate of HPV 16 was the highest, followed by HPV52, HPV58, HPV53 and so on.A total of 234 patients with positive biopsy results were found, and 77 patients with CIN2 or above lesions were detected.The ratio of CIN2 or above patients with HPV 16 was 11.8% (45/380), higher than 2.4% (32/1 328) of other high-risk subtypes such as HPV52 and HPV58.The difference was statistically significant(χ2=61.60, P < 0.001).In the age factor, the high-risk positive ratio of HPV in the 35-year-old group was 96.91% (1 225/1 264), higher than 94.38% (554/587) in the 35-year-old group.The difference was statistically significant (χ2=6.898, P=0.009).The detection rate of CIN1 disease was 12.64% (234/1 851) in HPV classification test, 8.59% (159/1 851) in liquid based thin layer cytology test (TCT), and the detection rate of TCT as a screening method was lower than that of HPV test method (χ2=16.01, P < 0.001). Conclusion The survey find that HPV subtypes are the most common in the order of HPV16, HPV52, HPV58, HPV53 and HPV18.HR-HPV infection was closely related to cervical lesions, and with the increase of high-risk infections, the degree of cervical lesions also increased.The detection rate of HPV typing is higher than that of TCT screening for cervical lesions.HPV genotyping has high sensitive and negative predictive value in screening cervical lesions.
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Objective To investigate the relationship between human papillomavirus (HPV) infection and cervical lesions, and to explore the application value of HPV typing in cervical disease screening. Methods Colposcopic examination and cervical biopsy were performed in 1 851 cases of HPV positive patients from January 2017 to September 2018 in the cervical disease clinic of Shanghai First People's Hospital.Pathological diagnosis was used as the gold standard to analyze the distribution of HPV subtypes and the relationship between HPV subtypes and cervical lesions. Results Among 1 851 patients, high-risk human papillomavirus (HPV) infection was the main type.The detection rate of HPV 16 was the highest, followed by HPV52, HPV58, HPV53 and so on.A total of 234 patients with positive biopsy results were found, and 77 patients with CIN2 or above lesions were detected.The ratio of CIN2 or above patients with HPV 16 was 11.8% (45/380), higher than 2.4% (32/1 328) of other high-risk subtypes such as HPV52 and HPV58.The difference was statistically significant(χ2=61.60, P < 0.001).In the age factor, the high-risk positive ratio of HPV in the 35-year-old group was 96.91% (1 225/1 264), higher than 94.38% (554/587) in the 35-year-old group.The difference was statistically significant (χ2=6.898, P=0.009).The detection rate of CIN1 disease was 12.64% (234/1 851) in HPV classification test, 8.59% (159/1 851) in liquid based thin layer cytology test (TCT), and the detection rate of TCT as a screening method was lower than that of HPV test method (χ2=16.01, P < 0.001). Conclusion The survey find that HPV subtypes are the most common in the order of HPV16, HPV52, HPV58, HPV53 and HPV18.HR-HPV infection was closely related to cervical lesions, and with the increase of high-risk infections, the degree of cervical lesions also increased.The detection rate of HPV typing is higher than that of TCT screening for cervical lesions.HPV genotyping has high sensitive and negative predictive value in screening cervical lesions.
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Objective: To optimize the synthesis process of 1-(4-aminophenyl)-5, 6-dihydro-3-(4-orpholinyl)-2(1H)-pyridinone.Methods: Using p-nitroaniline as the raw material, the target compound was obtained by amidation, cyclization, chlorination, nitration and reduction reactions. The sodium hydride catalyst for cyclization was taken out in the reaction, the reaction acid binding agent was improved, the reaction temperature of intermediate was reduced, the optimal ratio and reaction ratio condition were investigated, and the post-processing conditions were optimized for each reaction. Results: The target compound was confirmed by 1H NMR and 13C NMR data, and the total yield was 65.4%. Conclusion: The optimized process is simple to operate, mild and controllable, and the solvent is easy to recycle, low-polluting and more conducive to industrial production.
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Objective: To establish a quick method of ultra-performance liquid chromatography-quadrupole time-of-fight mass spectrometry (UPLC-Triple-TOF/MS) for the analysis of components of crude and sweated Dipsaci Radix. Methods: The separation was performed on the chromatographic column of Agilent Eclipse XDB-C18 (250 mm × 4.6 mm, 5.0 μm), and the mobile phase was 0.1% formic acid solution-methanol, with a gradient elution at a flow rate of 0.8 mL/min, the detection wavelength was 215 nm, the column temperature was 25 ℃. UPLC-Triple-TOF 5600+ time of flight liquid and mass spectrometer was used for mass spectrometry. Electrospray ion source negative ion mode was adopted, and the scanning range was m/z 100-1 500. The components of crude and sweated Dipsaci Radix were quickly identified according to the information obtained by high-resolution mass spectrometry combined with secondary mass spectrometry. Results: Fifty-two common components were identified or tentatively characterized based on the retention time and MS spectra. They were triterpenoid saponins, iridoids, phenolic acids etc. The crack rules of primary components were also analyzed. And comparing the components of crude and sweated Dipsaci Radix, it showed that the content of 20 components such as loganin acid, chlorogenic acid, loganin, isochlorogenic acid A, and asperosaponin VI was decreased after sweating, and caffeic acid, isochlorogenic acid, isochlorogenic acid C, and triplostoside A was increased. Conclusion: The types of components of crude and sweated Dipsaci Radix are identical, but there are differences in the content of the components. The content of the components of crude are higher than the sweated Dipsaci Radix. UPLC-Triple-TOF-MS technology was used to analyze the influence of “sweating” on the chemical composition of the Dipsaci Radix, so as to provide a theoretical basis for the study of the chemical constituents of sweated Dipsaci Radix and further research on the origin processing of Dipsaci Radix.
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To analyze the clinical characteristics of cases of novel coronavirus pneumonia and a preliminary study to explore the relationship between different clinical classification and liver damage. Consecutively confirmed novel coronavirus infection cases admitted to seven designated hospitals during January 23, 2020 to February 8, 2020 were included. Clinical classification (mild, moderate, severe, and critical) was carried out according to the diagnosis and treatment program of novel coronavirus pneumonia (Trial Fifth Edition) issued by the National Health Commission. The research data were analyzed using SPSS19.0 statistical software. Quantitative data were expressed as median (interquartile range), and qualitative data were expressed as frequency and rate. 32 confirmed cases that met the inclusion criteria were included. 28 cases were of mild or moderate type (87.50%), and four cases (12.50%) of severe or critical type. Four cases (12.5%) were combined with one underlying disease (bronchial asthma, coronary heart disease, malignant tumor, chronic kidney disease), and one case (3.13%) was simultaneously combined with high blood pressure and malignant tumor. The results of laboratory examination showed that the alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB), and total bilirubin (TBil) for entire cohort were 26.98 (16.88 ~ 46.09) U/L and 24.75 (18.71 ~ 31.79) U/L, 39.00 (36.20 ~ 44.20) g/L and 16.40 (11.34- ~ 21.15) mmol/L, respectively. ALT, AST, ALB and TBil of the mild or moderate subgroups were 22.75 (16.31- ~ 37.25) U/L, 23.63 (18.71 ~ 26.50) U/L, 39.70 (36.50 ~ 46.10) g/L, and 15.95 (11.34 ~ 20.83) mmol/L, respectively. ALT, AST, ALB and TBil of the severe or critical subgroups were 60.25 (40.88 ~ 68.90) U/L, 37.00 (20.88 ~ 64.45) U/L, 35.75 (28.68 ~ 42.00) g/L, and 20.50 (11.28 ~ 25.00) mmol/L, respectively. The results of this multicenter retrospective study suggests that novel coronavirus pneumonia combined with liver damage is more likely to be caused by adverse drug reactions and systemic inflammation in severe patients receiving medical treatment. Therefore, liver function monitoring and evaluation should be strengthened during the treatment of such patients.
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Objective To study the effect of small interfering ribonucleic acid (siRNA) silencing apoptosis signal-regulating kinase 1 (ASK1) on inflammatory response of lipopolysaccharide-induced alveolar epithelial A549 cells and its mechanism.Method Cell inflammation model of A549 cells was induced by lipopolysaccharide.The expression of ASK 1 in A549 cells was silenced by liposome transfection of siRNA.The mRNA and expression levels of ASK1,interleukin 6 (IL-6),interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-α) in A549 cells were detected by immunoblotting,real-time fluorescence quantitative polymerase chain reaction and enzyme-linked immunosorbent assay.Result The expression of IL-6,IL-8 and TNF-α in the experimental group was significantly higher than that in the control group (P<0.001),which indicated that the inflammatory model of A549 cells was successfully constructed.The mRNA level and expression of ASK1 in the interference group was significantly lower than that in the negative control group and the blank control group (P<0.01),indicating that silencing ASK1 was also successful.The expressions of IL-6,IL-8 and TNF-α in the interference group (0.37±0.04,0.32±0.04,0.48 ±0.13) were significantly lower than those in the negative control group (1.04±0.11,1.22±0.19,0.93±0.14) and the blank control group (1.01±0.14,1.01 ±0.23,1.02±0.25).The expression of IL-6,IL-8 and TNF-α protein in the interference group (pg/ml) (122.6± 11.0,537.2±42.4,159.2± 19.6) were also significantly lower than those in the negative control group (267.4±20.4,1 289.8±55.3,327.0±26.3) and blank control group (246.6±18.7,1 300.3±35.6,325.2± 18.3),with significant difference (P<0.05).There was no significant difference in each value between negative control group and blank control group (P>0.05).Conclusion Silencing ASK1 by siRNA can down-regulate the expression of IL-6,IL-8 and TNF-α in A549 cells,suggesting that ASK 1 may be involved in the regulation of lipopolysaccharide-induced inflammation in A549 cells.
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Objective: To investigate the effect and the possible mechanism of Rhizoma Paridis total saponin (RPTS) on human gastric cancer cell line MKN-45 proliferation, migration and invasion in vitro. Methods: MKN-45 cells were cultured in vitro and treated respectively with indicated concentrations of RPTS (2.5, 5.0, 10.0, 20.0, and 40.0 μg/mL) for 24 h, and cell viability of cell proliferation was detected by MTT assay; The invasive and metastatic ability of MKN-45 treated with indicated concentrations of RPTS (2.5, 5.0, 10.0 μg/mL) was detected by Transwell migration assay and wound healing assay; Elisa assay was employed to detect the concentrations of MMP-9 induced by LiCl after RPTS administration (10, 20, and 40 μg/mL) in the cell supernatant; Western blotting and qRT-PCR were respectively performed to investigate the invasion and migration related protein and mRNA level of VEGF, COX-2, and GSK-3β in RPTS-treated MKN-45 after LiCl stimulation for 24 h. Results: Compared with the control group, RPTS (10, 20, and 40 μg/mL) significantly inhibited the proliferation of MKN-45 cells (P < 0.05 and P < 0.001); RPTS (2.5, 5.0, 10.0 μg/mL) suppressed the invasion and migration of MKN-45 cells (P < 0.05 and P < 0.001); Compared with the model group, RPTS significantly downregulated the expression of MMP-9 in the cell supernatant of MKN-45 cells induced by LiCl (P < 0.05 and P < 0.01), and RPTS also decreased the protein and mRNA expression level of VEGF and COX-2, but it significantly upregulated the expression of GSK-3β at the protein and mRNA level (P < 0.05 and P < 0.001). Conclusion: RPTS play a pivotal role in suppressing the invasion and migration of MKN-45 cells in vitro, and its mechanism may be related to the regulating effects of the Wnt/β-catenin pathway in the human gastric adenocarcinoma cell.
ABSTRACT
Objective: To establish quantitative analysis of multi-components with single marker (QAMS) for determination of 10 phloroglucinol contents in effective fraction of Dryopteris fragrans. Methods: The relative correction factors of nine phloroglucinol (aspidin PB, aspidin AB, flavaspidic acid BB, saroaspidin A, flavaspidic acid PB, disflavapidic acid PB, flavaspidic acid AB, compound VI, and aspidinol B) were determined by HPLC method with the aspidinn BB as the internal standard, which were to calculate the content of each. At the same time, external standard method (ESM) was used to determine the contents of 10 components in effective fraction, and the differences between the two methods were compared to verify the feasibility and accuracy of QAMS method. Results: The relative correction factor (RCF) was good. There was no significant difference between the quantitative results with the two methods in the 12 batches of 10 phloroglucinols. Conclusion: The established QAMS method can be used for quantitative analysis of D. fragrans with aspidinn BB as the internal standard in the absence of reference substances, and provided a reference for the multi-index quality evaluation in effective fraction of D. fragrans.
ABSTRACT
OBJECTIVE: To study the correlation of common cardiovascular risk factors with brain iron deposition. METHODS: Eighty-four elderly subjects without neurological diseases or brain trauma were included in the study. The cardiovascular risk factors were comprehensively assessed. MRI examination was performed to obtain high-resolution T1-weighted images and enhanced susceptibility weighted angiography (ESWAN) images, and R2* figure was obtained by post-processing the ESWAN sequence. High definition T1 images were segmented using computer segmentation technique. After registration to the ESWAN image, R2* values of each region of interest were extracted. Multiple linear regression analysis was used to analyze the relationship of R2* values in each area of interest with gender, age and vascular risk factors. RESULTS Smoking was associated with increased R2* values in the hippocampus, white matter and cortex (β=0.244, 0.317, 0.277, P<0.05 or P<0.01). Hypertension was correlated with the increase of R2* in the putamen (β=0.241, P=0.027). Hyperglycemia was associated with the increase of R2* in the thalamus (β=0.234, P<0.05). In the thalamus, the R2* value of males was higher than that of females (β=0.320, P<0.05). Age was correlated with the R2* values of thalamus, caudate nucleus, pallidus, white matter and cortex (β=-0.218、-0.254、0.216、-0.280 and -0.238, P<0.05 or P<0.01). CONCLUSIONS Common cardiovascular risk factors may lead to iron deposition in the brain, and the deposition patterns vary with the gender, age and different risk factors.