ABSTRACT
ObjectiveThe human angiotensin converting enzyme2 （hACE2） transgenic mouse model was used to clarify the antiviral efficacy of BD-77 against a novel coronavirus SARS-CoV-2 and explore the action mechanism of BD-77 against SARS-CoV-2. MethodSARS-CoV-2 Omicron and Delta variant strains-infected VeroE6 cell models were established and administered with BD-77 to observe the antiviral effect of BD-77 in vitro. A kit was used to detect the effect of BD-77 in vitro on the binding of spike S protein of SARS-CoV-2 virus （Delta/Omicron） to angiotensin converting enzyme2 （ACE2）. Chromatography was adopted to detect the binding of BD-77 to the S protein and N protein of the novel coronavirus. hACE2 transgenic C57BL/6 mice were divided into a blank control group， SARS-CoV-2 infection group， BD-77 administration groups of 37.5 mg·kg-1 and 75 mg·kg-1， with eight mice in each group. The pneumonia model of SARS-CoV-2-infected hACE2 transgenic mice was built to observe the survival of the mice， detect the virus titer of the lung tissue of the mice， and observe the lesions in the lung tissue. ResultBD-77 had a certain inhibitory effect on Omicron and Delta variant strains in vitro， with median inhibitory concentration （IC50） of 526.3 mg·L-1 and 653.0 mg·L-1， respectively. BD-77 had no significant inhibitory effect on the binding of the S protein of WT， Omicron， and Delta variant strains of SARS-CoV-2 to ACE2 and had no binding effect with the S protein and N protein of the novel coronavirus. No mice in the blank group died， while the mortality rate of SARS-CoV-2-infected mice was 75%. There was a large amount of virus replication in the lung tissue of the mice and large areas of inflammatory infiltration in the lung tissue and interstitium. Compared with the model group， BD-77 administration groups of 37.5 mg·kg-1 and 75 mg·kg-1 could reduce the mortality of mice， significantly lower the virus titer in the lung tissue of mice （P<0.05）， and improve lung lesions. ConclusionBD-77 demonstrated significant inhibitory effects against SARS-CoV-2 virus in vitro and in vivo. However， its mechanism of action did not involve direct inhibition of the virus itself or intervention in the virus-host binding process. This finding suggests that the mechanism of action of BD-77 needs to be thoroughly investigated and elucidated by further experiments.
ABSTRACT
ObjectiveTo observe the therapeutic effect of BD-77 by nebulized inhalation on animal models of various respiratory viral infections and investigate the mechanism of broad-spectrum antiviral action of BD-77 using proteomics. MethodThe influenza virus H1N1/FM1 experiment used ICR mice and divided them into a normal group， model group， Tamiflu group， and BD-77 groups of 75 and 37.5 g·L-1 for inhalation of 20 min and 25 min. Human coronavirus 229E and OC43 experiment divided the BALB/c mice into a normal group， model group， chloroquine phosphate group， and BD-77 groups of 75， 37.5， 18.75， and 9.375 g·L-1， with 10 mice in each group. Influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 infection-induced pneumonia models were used to detect mouse lung index， and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the viral load in lung tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect related inflammatory factors in lung tissue， and proteomics analysis was performed on the lung tissue of OC43-infected mice. ResultCompared with that in the normal group， the lung index of mice in each infection group was significantly increased （P<0.01）， and viral nucleic acid could be detected in the lung tissue of mice infected with human coronaviruses 229E and OC43. The levels of interleukin-6 （IL-6）， IL-10， and tumor necrosis factor-α （TNF-α） in the lung tissue of mice infected with human coronavirus 229E were all significantly increased （P<0.01）. BD-77 could significantly reduce the lung index of mice infected with influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 （P<0.05， P<0.01）， cut down the viral load in the lungs of mice infected with human coronaviruses 229E and OC43 （P<0.01）， and lower the contents of IL-6， IL-10， and TNF-α in the lung tissue of mice infected with human coronavirus 229E （P<0.01）. Proteomics analysis of the lung tissue of OC43-infected mice showed that BD-77 regulated the AMPK signaling pathway， TNF signaling pathway， NOD-like signaling pathway， IL-17 signaling pathway， Forkhead box protein O （FoxO） signaling pathway， transforming growth factor-β （TGF-β） signaling pathway， and other signaling pathways. ConclusionNebulized inhalation of BD-77 is effective in treating pneumonia caused by influenza virus H1N1/FM1 and human coronaviruses 229E and OC43 infection in mice and may exert its antiviral effects by regulating the balance of cellular metabolism， enhancing the immune function of the host， and attenuating inflammatory responses.
ABSTRACT
In order to solve the problems of quality control and traceability of medical test lung for meeting the calibration conditions of JJF 1234-2018 Calibration Specification for Ventilators, the calibration device and method are researched for compliance and airway resistance of medical test lung in this paper. A calibration device for medical test lung is designed using constant volume active piston technology to simulate human breathing. Through comparison experiment, the deviation between this device and the similar foreign device can be found. The deviation is lower than 0.4% for lung compliance and lower than 0.7% for airway resistance. The calibration of lung compliance and airway resistance can be completed by this device. This device has a clear and complete traceability path to ensure quality control from the source. The calibration of ventilator is improved. This paper provides a reference for related metrology departments and medical institutions to study on quality inspection of respiratory medical instruments.
Subject(s)
Humans , Calibration , Ventilators, Mechanical , Respiration , Quality Control , LungABSTRACT
Objective:To investigate the auxiliary diagnostic value of serum midkine (MK) and thyroid stimulating hormone (TSH) for differentiated thyroid cancer (DTC).Methods:Seventy-one postoperative DTC patients (DTC group) treated with 131I were selected, and 143 patients with benign thyroid lesions (benign thyroid disease group) treated with surgery in Center Hospital of Xiaogan from March 2019 to December 2020 at the same period were also selected. Clinical data such as liver and kidney function indexes, positive rate of anti thyroglobulin antibodies (TGAb) and positive rate of thyroid peroxidase antibody (TPOAb) were collected before treatment, and their fasting blood samples were collected before treatment. Fully automated electrochemiluminescence immunoassay was used to measure free thyroxine (FT 4), free triiodothyronine (FT 3), TSH levels in patients′ serum. The serum MK levels were measured by enzyme-linked immunosorbent assay (ELISA). Binary Logistic regression model was used to screen for independent risk factors for the development of DTC. A receiver operating characteristic curve (ROC) was drawn to evaluate the efficacy of MK, TSH and MK combined with TSH, in aiding the diagnosis of DTC and its staging. Results:Serum TSH and MK levels in DTC group were higher than those in benign thyroid disease group: (3.55 ± 0.61) mU/L vs. (2.97 ± 0.46) mU/L, (394.25 ± 63.36) ng/L vs. (311.45 ± 42.66) ng/L, and the differences were statistically significant ( P<0.05). Elevated serum TSH and MK levels were independent risk factors for DTC. When MK combined with TSH was used to diagnose DTC, the area under the curve (AUC), sensitivity and specificity were higher than those of MK and TSH alone (0.925 vs. 0.859 and 0.783, 83.10% vs. 78.87% and 73.24%, 89.51% vs. 85.31% and 79.02%), and the differences were statistically significant ( P<0.05). Serum TSH and MK levels in stage Ⅲ and Ⅳ patients in DTC group were higher than those in stage Ⅰ and Ⅱ patients: (3.79 ± 0.65) mU/L vs. (3.42 ± 0.56) mU/L, (427.88 ± 52.73) ng/L vs. (311.45 ± 42.66) ng/L, and the differences were statistically significant ( P<0.05). The AUC, sensitivity and specificity of MK combined with TSH in the diagnosis of different stages of DTC were higher than those of MK and TSH alone (0.822 vs. 0.657 and 0.666, 73.90% vs. 56.52% and 56.52%, 83.33% vs. 77.08% and 79.17%), and the differences were statistically significant ( P<0.05). Conclusions:Serum TSH and MK levels are independent risk factors for the occurrence of DTC in patients, and the combination of them has certain auxiliary diagnostic value for the identification and staging of DTC.
ABSTRACT
Objective:To explore the identification value of carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC-Ag), cytokeratin 19 fragment (Cyfra211), thyroglobulin (TG), ferritin (Fer) and procalcitonin (PCT) in fine needle aspiration eluent in benign and malignant cervical nodules, and acquire the optimal diagnostic model.Methods:Three hundred and ninety-six single cervical nodule patients who underwent fine needle aspiration biopsy from August 2017 to August 2019 in the Center Hospital of Xiaogan City of Hubei Province were selected. The fine needle aspiration eluent levels of CEA, SCC-Ag, Cyfra211, TG, Fer and PCT were detected by electrogenerated chemiluminescence method. The results of cytopathological diagnosis were regard as "gold standard", and the diagnostic efficiency of single and combined indexes in fine needle aspiration eluent were analyzed by receiver operating characteristic (ROC) curve.Results:Among the 396 patients, malignant nodules was in 101 cases, and benign nodules was in 295 cases. The fine needle aspiration eluent levels of CEA, SCC-Ag, Cyfra211, TG and Fer in patients with malignant nodules were significantly higher than those in patients with benign nodules: (27.73 ± 10.63) μg/L vs. (16.81 ± 8.18) μg/L, (1.59 ± 0.74) μg/L vs. (1.09 ± 0.83) μg/L, (3.31 ± 1.48) μg/L vs. (1.66 ± 0.59) μg/L, (144.96 ± 38.93) μg/L vs. (95.03 ± 47.23) μg/L and (191.18 ± 80.13) μg/L vs. (137.87 ± 63.22) μg/L, the PCT was significantly lower than that in patients with benign nodules: (0.61 ± 0.24) μg/L vs. (1.01 ± 0.52) μg/L, and there were statistical differences ( P<0.01). The ROC curve analysis result showed that the CEA, Cyfra211 and TG had super diagnostic value (area under curve>0.7, Youden index>0.5); the area under curve of CEA, Cyfra211 combined with TG was significantly higher than other combined detection of 2 indexes ( P<0.05). Conclusions:The combined detection of CEA, Cyfra211 and TG in fine needle aspiration eluent can effectively distinguish the benign and malignant cervical nodules.
ABSTRACT
Objective To explore the effect of timosaponin B-II ( TB-II) on the differentiation of neural stem cells (NSCs) into tyrosine hydroxylase (TH) positive neurons in neonatal rats. Methods The biological functions of self-proliferation and multi-differentiation of NSCs were identified by primary culture, cell proliferation counting,morphological observation and immunology. NSCs of SD rats were cultured in vitro and treated with different concentrations of TB-II (10 μg/ml,30 μg/ml ,100 μg/ml) for 7 days. Immuno-histochemistry was used to detect the effect of TB-II on the differentiation of NSCs into TH-positive neurons, and Western blot was used to detect the expression of TH protein in neurons. Results ( 1) The cultured cells had the ability to self-proliferation,expressed nestin protein and differentiated into neurons and glial cells. So the cultured cells were conformed to the biological function of neural stem cells. (2)Compared with the control group,the TH positive cell ratio of TB-II 30 μg/ml group and TB-II 100 μg/ml group increased ((10. 03± 1. 36)%),( 20. 01± 3. 37)%),(31. 32± 3. 98)%) ,the difference was significant ( t=6. 15, 16. 54,both P<0. 05). There was no significant difference between TB-II 10 μg/ml group and control group (P>0. 05). (3)Western results showed that the relative expression of TH protein in TB-II 30 g/ml group and TB-II 100 μg/ml group was higher than that in control group,the difference was statistically significant (con-trol group: (1. 02±0. 24),TB-II 30μg/ml group: (3. 64±1. 78),TB-II 100 μg/ml group: (5. 88±2. 34);t=12. 58,9. 15,both P<0. 05). There was no significant difference between TB-II 10 μg/ml group and con-trol group (P>0. 05). Conclusion TB-II can promote the differentiation of NSCs into TH-positive neurons.
ABSTRACT
As an important clue and reference for modern compound medicine,Chinese medicine compatibility has become an innovative research and development mode of fixed dose combination with Chinese characteristics and traditional Chinese medicine (TCM) characteristics.However,it is difficult to find and determine the effective substances because of the complex composition,the huge amount and the structure diversity.In order to solve these problems,innovative ideas,methods and techniques are needed.
ABSTRACT
[ ABSTRACT] AIM:To investigate the therapeutic effects of lentivirus-mediated shRNA targeting growth hormone secretagogue receptor 1a (GHSR1a) on colorectal cancer cell line SW480 both in vitro and in vivo.METHODS: Human GHSR1a sequence was used for the design of shRNA targeting GHSR1a, which was introduced to lentivirus, followed by transfection into SW480 cells.CCK-8 assay was performed to detect cell viability.The mRNA expression of GHSR1a and PTEN in colorectal cancer cells was detected by RT-PCR.The protein levels of GHSR1a, ghrelin, PTEN, p-AKT and p53 were determined by Western blot.Stable GHSR1a silencing in SW480 xenografts in nude mice was established.After the mice were sacrificed and weighted, immunohistochemistry was used to detect the positive expression of Ki-67 and PTEN in the tumors.RESULTS:GHSR1a was over-expressed in the malignant cells in comparison with the normal cells in vitro. The tumor specific lentivirus-mediated shRNA targeting GHSR1a gene and GHSR1a knockdown SW480 cells were success-fully constructed.After transfection with GHSR1a shRNA, the expression of GHSR1a at mRNA and protein levels was markedly inhibited in the SW480 cells.Furthermore, GHSR1a silencing by specific shRNA showed increased PTEN, inhi-bition of AKT phosphorylation and promotion of p53 release in the SW480 cells.In vivo results demonstrated that down-re-gulation of GHSR1a in the SW480 cells significantly decreased the expression of Ki-67 and increased the expression of PTEN in the tumor tissues, leading to a marked reduction in tumor weight in comparison to blank control or negative control tumors.CONCLUSION:Down-regulation of GHSR1a by shRNA technique inhibits the growth of colorectal cancer cell line and xenograft tumor through activation of the PTEN/PI3K/AKT signaling pathways.
ABSTRACT
Objective To analyze influence factors of serum A-TG level after DTC 131 I treatment ,to explore whether A-TG can be used as an indicator of follow-up ,recurrence and metastasis of DTC .Methods From 2008 January to 2013 February ,a total of 106 DTC patients underwent operation excisions of thyroid radioiodine were treated with 131 I .Before and 1 month after treatment , the levels of A-TG ,TG ,THS were measured .The relationship of A-TG and thyroid operation range operation times ,thyroid gland residual volume and time ,the levels of TSH and TG ,thyroid cancer metastasis and recurrence were examined .Results Serum A-TG concentration associated with operation scope ,frequency(P<0 .05) .It was showed that the A-TG concentration was positively correlated with the metastasis of thyroid cancer .Conclusion The serum TG level detection combined with A-TG and 131 I-WBS can improve the diagnostic sensitivity and accuracy of DTC recurrence and metastasis .
ABSTRACT
Objective: To investigate the influence of Acorus gramineus (Soland), a crude extract, SCP01, and a purified component, SCP02, and of Rosmarinus officinalis L., X0728 on human mast cells (HMC-1 Cell Line). Methods: Current-voltage of P2X7 receptors on human mast cell membrane activated by ATP was recorded by the whole-cell patch clamp technique. Results: The current at-100 mV mediated by P2X7was inhibited by (27.6±2.0) % in the presence of 40 μg/mL SCP01 and by (29.5±2.2) % in the presence of 40 μg/mL SCP02, which was identified as α-asarone. 42 μg/mL of the commercially available α-asarone inhibited the P2X7-mediated current by (52.2±2.0) %. In contrast to SCP01 and SCP02, 40μg/mL X0728 provoked stimulation of the current by (28.6±2.8) %. All effects were voltage-independent. Conclusion: The inhibition of P2X7by α-asarone will inhibit intracellular calcium increase and this may account for the inhibition of reported excitotoxic cell death. The pharmacological function of P2X7stimulation by X0728 needs further investigation.
ABSTRACT
Objective: α-asarone is a major effective component that can be isolated from Acorus tatarinowii Schott,a Chinese herbal medicine. Clinical investigations have shown that α-asarone has strong sedative and anti-convulsive action in the central nervous system. In recent years, several medicines containing a-asarone were applied in treatment of asthma, bronchitis, expectorant, or epilepsy. However, the underlying cellular mechanism of ct-asarone is still unknown. Here the authors considered EAAC1, the transporter for the excitatory glutamate, as a possible target. Methods: Supercritical CO2 fluid extraction and silica gel column chromatography were used to obtain ct-asarone from the rhizomes of Acorus tatarinowii Schott. Xenopus oocytes with heterologously expressed EAAC 1 were used as a model system. Rate of glutamate uptake was measured by means of isotopic tracer technique. Glutamate-induced current was recorded under two-electrode voltage clamp. 40μg/mL of ct-asarone was used for testing its effect on EAAC1 activity. Results: ct-asarone induced a slight, but still significant stimulation of rate of glutamate uptake by 15%. In contrast, EAACl-mediated current became reduced (by 30% at -100 mV). Since EAAC 1 can operate in transport and also in an ion-channel mode, the result indicates strong inhibition of the channel mode. This inhibition is voltage-dependent becoming larger at more negative potentials. Conclusion: The stimulation of glutamate uptake reduces glutamate concentration in the synaptic cleft and, hence, reduces excitatory synaptic activity. The inhibition on the ion-channel mode stabilizes the membrane potential, and therefore, also contributes to reduced excitatory activity.
ABSTRACT
Objective To study the influence of aldehydes and tannic acid on soft capsules, approach how to prevent soft capsules from delayed disintegration. Methods Add aldehydes and tannic acid to soft capsules material ingredient, to proceed accelerating stability trial and test the disintegration time. Result Little increase of aldehydes amount had great effect on the disintegration time of soft capsules, different aldehydes and tannic acid had different effect on the disintegration time, adding some organic acid such as fumaric acid to the material would improve disintegrating property. Conclusion The amount of aldehydes and tannic acid contained in the material and ingredient should be controlled strictly when prepare soft capsules so that the disintegration time of capsules could be complying with the Pharmacopeia.
ABSTRACT
Objective To study the chemical constituents from Dipentodon sinicus. Methods Isolation and purification were carried out on silica gel column and Sephadex LH-20, the structures were elucidated on the basis of chemical evidences and spectral analysis. Results Nine compounds were isolated and their structures were identified as n-1-triacontanol (Ⅰ), ?-sitosterol (Ⅱ), pyrocatechol (Ⅲ), p-dihydroxyl benzene (Ⅳ), coniferyl aldehyde (Ⅴ), vanillic acid (Ⅵ), ergosterol peroxide (Ⅶ), ?-sitosterol-3-O-?-D-glucosides-3'-O-tritriacontanate (Ⅷ), and daucosterol (Ⅸ). Conclusion All the nine compounds are isolated from the title plant for the first time, among them, compound Ⅷ is a new compound named as dipentosinin.