Molecular Mechanism of "Transmission Between Lung and Brain" of Influenza and Intervention Effect of Maxing Shigantang Based on JAK1/STAT1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the molecular mechanism of "transmission between the lung and brain" of influenza based on Janus kinase 1/signal transducer and activator of transcription 1（JAK1/STAT1） signaling pathway and further investigate the intervention effect of Maxing Shigantang （MXSGT）. MethodA total of 100 SPF BALB/c mice were randomly divided into a normal group，a model group，an oseltamivir group （21.63 mg·kg-1·d-1），an antiviral granules group（3.9 g·kg-1·d-1）， and an MXSGT group（6.05 g·kg-1·d-1）， with 20 mice in each group. The pneumonia model was induced in mice except for those in the normal group by intranasal infection of influenza A virus（IAV）. Twenty-four hours after modeling，mice were treated with corresponding drugs， while those in the normal group and the model group received the same amount of normal saline by gavage， once a day for 3 and 7 days. The pathological changes in the lung and brain were observed by hematoxylin-eosin（HE）staining. The mRNA expression of IAV nucleoprotein（NP），JAK1， and STAT1 in the lung and brain was detected by real-time quantitative polymerase chain reaction（Real-time PCR）， and the protein expression of JAK1 and STAT1 in the lung and brain was detected by Western blot. Immunohistochemical method was used to detect the expression of phosphorylated（p）-STAT1 in the lung and brain tissues， and enzyme-linked immunosorbent assay（ELISA） was used to detect the serum levels of interleukin-1β（IL-1β） and interleukin-10（IL-10）. ResultCompared with the normal group， the model group showed obvious pathological changes in the lung tissues and cerebral cortex， increased relative mRNA expression of IAV NP in the lung （P<0.01）， elevated mRNA and protein expression of JAK1 and STAT1 in the lung and brain tissues （P<0.05，P<0.01），up-regulated expression level of p-STAT1 in lung tissues and cerebral cortex （P<0.05，P<0.01）， and increased serum level of IL-1β （P<0.05）. Compared with the model group， the MXSGT group showed alleviated pathological damage to lung tissues and cerebral cortex， decreased relative mRNA expression of IAV NP in lung tissues（P<0.01），reduced mRNA and protein expression levels of JAK1 and STAT1 in lung tissues and brain tissues（P<0.05，P<0.01）， and increased serum level of IL-10（P<0.01）. ConclusionThe abnormal activation of the JAK1-STAT1 signaling pathway may be one of the molecular mechanisms of "transmission between the lung and brain" of influenza. As an effective compound prescription against the influenza virus，MXSGT can alleviate the pathological damage of brain tissues in mice infected with IAV by regulating the level of cytokines mediated by this pathway.

Pneumocystis jirovecii pneumonia induced by low-dose methotrexate in a patient with chronic urticaria

Abstract Methotrexate has immunosuppressive effects and is administered for refractory chronic urticaria. We present a case of Pneumocystis jirovecii pneumonia in a patient with refractory chronic urticaria managed by low-dose weekly methotrexate treatment (total cumulative dose 195mg). Our study highlights the importance of providing prompt diagnosis and treatment of Pneumocystis jirovecii pneumonia in patients with chronic urticaria under methotrexate therapy.

Protective Effects of Cornus Officinalis Total Glycosides and Cornus Polysaccharides on Myocardial Mitochondria of Acute Myocardial Infarction Rats: an Experimental Study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Cornus Officinalis total glycosides (COTG) and Cornus polysaccharides (CP) on myocardial mitochondria and expression levels of glycogen synthase kinase-3β (GSK-3β) of acute myocardial infarction (AMI) rats.</p><p><b>METHODS</b>The AMI rat model was established by ligating the left anterior descending branch of coronary artery. Rats were divided into 5 groups according to random digit table, i.e., the sham-operation group, the model group, the COTG prevention group, the CP treatment group, the COTG treatment group, 12 in each group. Normal saline was administered to rats in the normal control group and the model group by gastrogavage. Corresponding medication was respectively administered to rats in the rest 3 groups by gastrogavage. The cardiac function was detected by echocardiography and hemodynamics. The infarct size was determined by Masson trichrome staining. The expression of mitochondrial biogenesis genes such as a subunit of peroxisome proliferators-activated receptor-γ coactivator-1 (PGC-1α), PGC-1β, nuclear respiratory factor-1 (NRF-1), and GSK-3P mRNA were detected by Real-time PCR.</p><p><b>RESULTS</b>Compared with the sham-operation group, the myocardial infarction size increased, cardiac function decreased, the expression of PGC-1α, PGC-1β, and NRF-1 mRNA decreased, and the expression of GSK-3β mRNA increased (all P <0. 05). Compared with the model group, myocardial infarction sizes were reduced, cardiac function was improved, the expression of NRF-1 mRNA was elevated in the COTG prevention group, the CP treatment group, the COTG treatment group; the expression of the PGC-1α and PGC-1β mRNA was elevated in the COTG prevention group and the CP treatment group; the expression of GSK-3β mRNA was reduced in the CP treatment group (all P <0. 05). Compared with the CP prevention group, fractional shortening (FS) and aortic systolic blood pressure (SBP) increased in the CP treatment group; ejection fraction (EF) decreased in the CP treatment group; the expression of PGC-1α, PGC-1β, NRF-1 mRNA were reduced in the the CP treatment group and the COTG treatment group; the expression of GSK-3β mRNA decreased in the CP treatment group (all P <0. 05). Compared with the COTG treatment group, FS, EF, left ventricular end systolic pressure (LVESP), SBP, and the expression of GSK-3β mRNA were reduced in the CP treatment group (P <0. 05).</p><p><b>CONCLUSIONS</b>COTG and CP could improve cardiac function, reduce the myocardial infarction area, and promote biogenesis of myocardial mitochondria. Their protective effects on the mitochondria of cadiocytes might be achieved by GSK-3β signalina pathway.</p>

Differentiation of rat bone marrow-derived mesenchymal stem cells into cardiomyocyte-like cells induced by cyclic stretching strain / 生物医学工程学杂志

Bone marrow-derived mesenchymal stem cells (BMSCs) are multipotent stem cells that differentiate into a variety of cell types and widely used in tissue regeneration engineering. The purpose of this study is to investigate whether the cyclic biaxial stretching strain could promote the rat BMSCs (rBMSCs) to differentiate into cardiomyocyte-like cells in vitro. The second or third generation of rBMSCs were randomly divided into the cyclic stretching stain group, the control group and the blank group. Those rBMSCs in the cyclic stretching strain group were seeded on a silicone membrane with complete medium were exposed to biaxial stretching strain of 10% of membrane at a frequency of 1 Hz lasting for 6 h, 12 h and 24 h. Those in the control group were seeded on silicone membrane with complete medium. Those in the blank group were seeded in the 6-wells plates with complete medium. The mRNA expression of GATA4 and myocyte-specific enhancer factor 2C (MEF-2C) were detected by the real-time fluorescent quantification PCR and the protein expression of connexin 43 (Cx43) was detected by using the Western blot method. The results showed that the mRNA expression level of the GATA4 and MEF-2C, and the protein expression level of Cx43 were significantly higher in the cyclic stretching strain groups, compared with those in the relative control groups (P < 0.05). It suggests that cyclic biaxial stretching strain could play a part in the induction of rBMSCs to differentiate into cardiomyocyte-like cells in vitro, but the differentiation mechanism is still unclear.

Inhibition of Jumi extraction on growth of human cervical cancer cell line HeLa / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the inhibition of Jumi (traditional Chinese medicine) extraction on the growth of human cervical cancer cell line HeLa.</p><p><b>METHODS</b>Nude mouse model of human cervical cancer HeLa cell transplantation was established. The nude mice bearing cancer were randomly divided into control group and Jumi treated groups with different concentration (0.001, 0.002, 0.005, 0.01 mg/ml). The growth of cervical cancer cell in experimental mice were measured. Cultured HeLa cells were incubated in culture media with or without Jumi extract for 48 hours. Cell proliferation rate, cell apoptosis, caspase-3/7 and caspase-6 activity were determined by MTT colorimetric assay, flow cytometry analysis and spectrophotometric detection, respectively.</p><p><b>RESULTS</b>With the increase of the concentration of Jumi extract, tumor-bearing mice tumor inhibition rate gradually increased. The proliferation of cultured HeLa cells were significantly inhibited by Jumi extract in a dose-dependent manner. IC50 was 0.004 mg/ml. Apoptosis rates in the cells treated with Jumi extract were higher than those of the control group. Compared with the control group, except for lower Jumi treated group (0.001 mg/ml), caspase-3/7 and caspase-6 activity were significantly increased in the all Jumi treated groups.</p><p><b>CONCLUSION</b>Jumi extract can inhibit the proliferation of human cervical cancer cell line HeLa in vitro in a dose-dependent manner and promote cell apoptosis through caspase-3, caspase-7 and caspase-6 pathway.</p>

Effects of electrical stimulation on the differentiation of mesenchymal stem cells into cardiomyocyte-like cells / 生物医学工程学杂志

The aim of this study is to investigate the effects of electrical stimulation ES) on the induction of rat bone marrow mesenchymal stem cells (rBMSCs) to differentiate into cardiomyocyte-like cells in vitro. The third or fourth-generation of the rBMSCs was randomly divided into three groups, i. e. ES group, 5-Azacytidine (5-Aza) group, and control group. Those in the ES group with complete medium were exposed to 1,2,4 and 6V, 2Hz, 5ms ES for 2h everyday,lasting for 10d. Those in the 5-Aza group were induced by 10 micromol/L 5-Aza for 24h, then the medium was changed to complete medium without 5-Aza. Those in the control group were only cultured with complete medium. The growth status and morphological features of rBMSCs were observed by inverted phase microscope. The mRNA expressions of GATA4, a-actin, ACTN2 and TNNT2 were determined by Real-time fluorescent quantification PCR, and the protein expression of TNNT2 was detected with immunofluorescence staining. The results showed that the mRNA expression level of the GATA4, a-actin, ACTN2 and TNNT2 and the protein expression level of the TNNT2 were significantly higher in the ES group and 5-Aza group, compared to those in the control group(P<0. 05). It suggested that ES could induce rBMSCs differentiation into cardiomyocyte-like cells in vitro.

Experimental study in the effect of various intracranial hypertension upon the cerebral blood flow by transcranial Doppler / 中华超声影像学杂志

Objective To investigate the changes of transcranial Doppler (TCD) patterns and parameters in various intracranial hypertension.Methods Sixty rabbits were randomized into 3 groups:control group,the group of mild-to-moderate intracranial hypertension,the group of serious intracranial hypertension.Acute intracranial hypertension was induced by inflating the balloon inserted into the epidural space.Blood flow velocity was measured with TCD and intracranial pressure (ICP) as well as cerebral perfusion pressure(CPP) was measured.Mannitol was injected to the animals of intracranial hypertension,blood flow velocity and ICP was measured.ResultsTwo characteristic flow patterns vere observed in the group of mild-to-moderate intracranial hypertension:high resistance pattern,systolic flow.At the last stage of extreme intracranial hypertension in the group of serious intracranial hypertension Doppler sonograms showed three characteristic flow patterns in the following sequence:retrograde diastolic flow,very small systolic flow and zero flow.Multiform retrograde diastolic flows were related to the phases of brain death.Very small systolic flow showed three shapes:systolic-spike,small systolic triangular and small double peak.AftermedicinaltreatmentICPdecreasedinthethegroupofmild-to-moderateintracranial hypertension,blood flow velocity also increased.After the same treatment IC,P and blood flow velocity did not change in the group of serious intracranial hypertension.ConclusionsAnalysing patterns and parameters of TCD may be helpful for evaluating ICP indirectly as well as clinical experience.

Low frequence pulsed electromagnetic fields induce chondrocyte-like cells differentiation of rat bone marrow-derived mesenchymal stem cells in vitro / 生物医学工程学杂志

Mesenchymal stem cells (MSCs) are multipotent stem cells that differentiate into a variety of cell types. Low frequency pulsed electromagnetic fields (LFPEMFs) therapy can causes biochemical changes at the cellular level to accelerate tissue repair in mammals. So, we tested the hypothesis that LFPEMFs can promote chondrogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBMSCs) in vitro. The rBMSCs were isolated by adherence method and the third-generation of the rBMSCs were randomly divided into LFPEMFs groups, chondrocyte-induced group and control group. LFPEMFs groups with complete medium were exposed to 50Hz, 1mT PEMFs for 30 min every day, lasting for 10, 15 and 20 d, respectively. Chondrocyte-induced group were treated with chondrogenic media, while control groups were only cultured with complete medium. The mRNA expressions of type II-collagen (Col II) and aggrecan were determined by Real-time fluorescent quantitation PCR. The protein expression of Col II and aggrecan were detected with toluidine blue stain or immunocytochemical stain, respectively. The result showed that the mRNA and protein expression level of Col-II and aggrecan were significantly higher in the LFPEMFs group or chondrocyte-induced group, compared to the control group. It suggest that LFPEMFs could contribute to rBMSCs to differentiate into chondrogenic differentiation in vitro.

Separation and purification of catalpol from leaves of Rehmannia by macroporous adsorption resins:a priliminary study / 军事医学科学院院刊

Objective:To obtain the optimal conditions for separating catalpol from leaves of Rehmannia by selecting appropriate macroporous adsorption resins.Methods:The detection indication was the content of catalpol, which was determined by HPLC method. Twelve different kinds of macroporous adsorption resins were studied on the static capacity of adsorption and desorption, and H103 resin was selected for the research of separation and purification.Results:The H103 resin had a good capacity for adsorption and desorption.The best process of purifying catalpol by H103 resin was 1mg/ml concentration, the adsorption rate of 1-2 BV/h,the flow rate of 1-3 BV/h, and 8 BV with 10% alcohol.Conclusion:The method is simple and available, which can simplify the production process and lower costs.

Effect of iron on vasoconstriction in the isolated rat aorta / 生理学报

The present study was to examine the effect of iron on isolated rat aortic rings, and to elucidate the underlying mechanism. The thoracic aortic rings without endothelium of male Sprague-Dawley rats were mounted on a bath system. Isometric contractions of aortic rings were measured. The results obtained are as follows. (1) Ferric ammonium citrate (FAC) (100 micromol/L) caused a phasic response with an initial transient contraction followed by a relaxation in thoracic aortic ring. The maximal contractile amplitude was 24.02+/-2.37% of the maximal contraction induced by KCl, the duration of phasic contraction lasted for about 20 min. (2) In high Ca(2+) Krebs-Henseleit (K-H) solution (twice of the normal concentration), the contractile amplitude induced by FAC was enhanced. After the aortic rings were incubated with nifedipine for 15 min to block the L-type Ca(2+) channel, the iron-induced contraction was attenuated. (3) In Ca(2+)-free K-H solution, addition of FAC caused a strong and sustained contraction in the presence of PDBu. (4) Pretreatment of FAC for 30 min decreased the KCl-induced contraction and also caused a significant reduction in the contractile response to phenylephrine (PE). Pretreatment of the arteries with DMSO, catalase or glutathione before FAC exposure prevented the decrease in contraction responses to PE (P<0.05). It is therefore concluded that iron causes phasic contraction of vascular smooth muscle, in which both extracellular Ca(2+) entry through L-type Ca(2+) channel and increase in Ca(2+) sensitivity of smooth muscle cells are involved. Exposure to iron causes inhibitory effects on KCl- or PE-induced contractions in isolated thoracic arteries. Reactive oxygen species and glutathione may be involved in iron-induced contraction dysfunction.

Inhibitory effect of iron on vasodilatation in the isolated rat aorta / 中国病理生理杂志

AIM: The objectives of the present study were to examine the effect of iron on relaxation of isolated rat aortic rings,and to elucidate the underlying mechanism. METHODS: The thoracic aortic rings of male Sprague-Dawley rats were mounted on bath system. Vasodilatation of aortic rings preconstricted with 10 -6 mol/L of phenylephrine (PE) was measured. RESULTS: (1) Exposure of endothelium-intact aortic rings to ferric ammonium citrate (FAC) for 30 min caused a significant reduction in the relaxation response to acetylcholine (ACh). Pretreatment with L-arginine (L-Arg) before incubation with FAC did not reverse the inhibition of relaxation response to ACh completely. (2) In endothelium-intact aortic rings,L-Arg relaxed the PE preconstricted vessels. Exposure to FAC for 30 min caused the decrease in the relaxation response to L-Arg. There was no difference in the relaxation response to nitric oxide donor,sodium nitroprusside, between endothelium-denuded arteries treated with or without FAC. (3) Dimethyl sulfoxide had no effect on the inhibition of relaxation to ACh by FAC in endothelium-intact rings. Pretreatment of arteries with glutathione and catalase prevented the decrease in relaxation responses to ACh induced by FAC. (4) The nitric oxide synthase activity was (56.49?2.49)?10 3U/g protein in normal aorta with endothelium,while after incubation with FAC for 30 min,it reduced to (25.15?5.75)?10 3U/g protein ( P