Aim To investigate the effect of bufalin on proliferation and expression of WT1 in K562 cells. Methods The colony number of K562 cell was detec-ted with semi-solid culture assay. The cell cycle was measured by flowcytometry, and the expression of WT1 was observed with immunocytochemistry. Subcutaneous tumor models established by K562 cells in BALB/C nu/nu mice were divided into three groups, including model group, bufalin group and positive control group. After 21 days, the subcutaneous tumors were removed for calculating the inhibitory rate of tumor growth. HE staining and immunohistochemistry were used to ob-serve the morphological changes and the expression of WT1 . Results ① Bufalin could significantly decrease the colony number of K562 cell, arrest it at G0/G1 phase and down-regulate its expression of WT1 in a dose-dependent manner. ② Compared with the model group, the tumor inhibitory rate was much higher, while the volume and the weight were obviously lower in the other two groups. ③Bufalin could induce apop-tosis, necrosis, hemorrhage and fibrosis with HE stai-ning, and down-regulate the expression of WT1. Con-clusion Bufalin could inhibit the proliferation, arrest the cell cycle at G0/G1 phase and down-regulate the expression of WT1 in vitro. Bufalin could inhibit the tumor inhibitory rate, the volume and the weight of the subcutaneous tumors, induce apoptosis, necrosis, hemorrhage and fibrosis with HE staining and down-regulate the expression of WT1 .

Objective To investigate the effect of normal saline(NS)of different temperature on morphological changes and nitric oxide synthase(NOS)expression of spinal cord.Methods The spinal canal of 96 SD adult rats was opened at T9.which of 24 rats was flushed with 37 ℃ NS,24 with 20 ℃ NS,24 with 4 ℃ NS respectively,and which of 24 rats in control group received no flushing.The spinal canal was closed one hour later,and the spinal cord was taken out 24 hours later.Then the water content in spinal cord was determined by dry-wet method.The morphological changes of spinal cord were observed under light microscope and the electronic microscope.The amount of NOS-positive neuron was measured by ?-NADPH histochemical methods.Results The water content in spinal cord was(66.53?0.61)% in control group,(66.75?1.00)% in 37 ℃ group,(70.55?0.77)% in 20 ℃ group,(71.92?2.50)% in 4 ℃ group.The spinal cord of control group and 37 ℃ group contained less water than that of 20 ℃ group and 4 ℃ group.There were no obvious morphological changes in the control group and 37 ℃ group.In 20 ℃ and 4 ℃ groups,the demyelination of axon,swelling of cell body and the disappearance of tigroid body were observed under light microscope,the partial disaggregation of medullary sheath,swelling of mitochondria and disappearance of mitochondria crista could be observed under electron microscope.The amount of NOS-positive neuron in spinal cord was(18.75?2.12),(18.63?1.41),(14.75?1.67),(8.13?1.25)in control,37 ℃,20 ℃ and 4 ℃ groups,respectively.The control group and 37 ℃ group showed more NOS-positive neuron than those of 20 ℃ group and 4 ℃ group.Conclusion NS below 20 ℃ can injury spinal cord.It is suitable to choose 37 ℃ NS to flush brain and spinal card during operation.

Objective · To investigate the effects of insulin on high glucose-cultured humanmononuclear cell line THP-1 and macrophage phenotype transformation in diabetic wounds. Methods · THP-1 cells were cultured with normal (5.6 mmol/L) and high (25 mmol/L) glucose, respectively,stimulated with PMA for differentiation, and induced to M1 macrophages with LPS. After treated with insulin for 6 h, expression changes of M1 type macrophage markers inducible nitric oxide synthase (iNOS), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β), as well as M2 type macrophage markers arginase1 (Arg1) and IL-10 were detected using real-time PCR andWestern blotting. High fat diet feeding plus multiple intraperitoneal injections of low dose streptozotocin (STZ) were used to induce type II diabetes rat model. After blood glucose level has been stable for five weeks, two fullthickness skin wounds with the diameter of 1cm were made on the back of DM rats. Wounds were randomly assigned to being treated with insulin (0.2 U insulin /20 μL saline) or saline (20 μL saline) using the random number table. Characteristics of macrophagephenotypes were observed 3, 7, and 25days after wounds were made. Normal rats (n=3) served as controls. Results · After being cultured with high glucose, the mRNA levels of M1 markers iNOS and TNF-α were up-regulated in LPS-induced THP-1 cells, while the mRNA levels of M2 markers Arg1 and IL-10 were down-regulated.Afterbeing treated with insulin for 6 h, mRNA levels of iNOS and TNF-α weredown-regulated, protein levels of iNOS, IL-1β were down-regulated too, while mRNAand protein levels of Arg1 and IL-10 were up-regulated. In addition, the expression level of phosphorylated NF-κB-p65 was significantly increased after high glucose culture and was significantly decreased after insulin intervention. Compared to normal rat skin wounds, the expression of iNOS in macrophages was significantly increased in wounds of diabetic rats. The expression of iNOS in macrophages was high in saline treated wounds 3 and 7 days after the wounds were made and the expression of Arg1 was low 25 days after the wounds were made. In insulin treated wounds, the expression of iNOS started to decrease on day 7 after the wounds were made and the expression of Arg1 was significantly higher than that in saline treated wounds on day 25 after the wounds were made. Conclusion · Insulin can induce macrophage phenotype transformation from M1 to M2 under high glucose condition and the mechanism may be associated with the phosphorylation of NF-κB-p65.

The ITS/ITS2 barcodes were used to simply and effectively identify Codonopsis Radix and its adulter-ants. In this study, ITS (internal transcribed spacer of unclear ribosomal DNA) regions were amplified using PCR (polymerase chain reaction) from thirty-three samples of Codonopsis Radix and ITS2 regions were obtained from the ITS sequences using the hidden Markov model (HMMer)-based annotation methods. The sequences of ITS/ITS2 regions were aligned and the genetic distances were computed by MEGA5.0. Species identification efficiency of ITS/ITS2 sequences were evaluated using BLAST1 and nearest distance methods. The results indicated that The sequences lengths of ITS regions of Codonopsis Radix were 654-655 bp, and the lengths of ITS2 regions were 239 bp. The intraspecific genetic distances among Codonopsis Radix were obviously lower than the interspecific genetic distance between Codonopsis Radix and its adulterants. Therefore, ITS/ITS2 regions can stably and accu-rately distinguish Codonopsis Radix and its adulterants.

Objective To investigate the necessity of low-intensity anticoagulation standard in patients after heart valve replacement and the rationality of INR in our hospital.Methods 681 eligible candidates were anticoagulated under the current guidelines for postoperative anticoagulation therapy in our hospital(AVR 1.5-2.0,MVR 2.0-2.5,DVR 2.0-2.5,TVR 2.5-3.0).We monitored the patient 's PT regularly and analyzed the occurrence of anticoagulation-related complications,such as bleeding,thrombosis and embolism.Results 602 cases completed the follow-up.During the period of follow-up,66 patients had bleeding tendencies,the incidence of bleeding complications was 10.96％ (66/602).1 1 patients had embolism complications,the incidence of thrombotic complications was 1.83 ％ (11/602).The average of INR was 2.24± 0.68,the mean oral Warfarin dose was(3.12± 1.14) mg/d.Conclusion Our study suggest that the effect of low-intensity anticoagulation after heart valve replacement is reliable.Further more,the current anticoagulation standards of our hospital meet the requirements of postoperative clinical anticoagulant after heart valve replacement in our region.

OBJECTIVE:To establish a method for simultaneous determination of 4 psoralen compounds in Buwu tincture. METHODS:HPLC was performed on the column of Dikma Diamonsil C18 with mobile phase of acetonitrile-0.2% Acetic acid by gradient elution at flow rate of 1 ml/min,detection wavelength was 246 nm,column temperature was 30 ℃,and the injection vol-ume was 15 μl. RESULTS:The linear range was 13.00-208 μg/ml for angelicin,26.00-416 μg/ml for bavachin,24.50-392 μg/ml for psoralidin and 37.88-606 μg/ml for isobavachalcone,respectively(r≥0.999 6);RSDs of precision,stability and reproducibility tests were less than 2.00%;recoveries were 95.22%-97.23%(RSD=0.87%,n=6),100.24%-104.64%(RSD=1.62%,n=6), 102.28%-104.39%(RSD=1.47%,n=6)and 97.68%-100.17%(RSD=0.97%,n=6),respectively. CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Buwu tincture.

The 2019 novel coronavirus (2019-nCoV) outbreak in Wuhan, Hubei Province, China in 2019 led to large numbers of peoplebeing infected and developing atypical pneumonia (coronavirus disease 2019, COVID-19). Typical imaging manifestations ofpatients infected with 2019-nCoV has been reported, but we encountered an atypical radiological manifestation on baselinecomputed tomography (CT) images in three patients from Wuhan, China infected with the 2019-nCoV. Surprisingly, the onlysimilar CT finding was a solitary sub-centimeter ground-glass nodule adjacent to bronchovascular bundles, which could beeasily overlooked. In addition, the follow-up images in these patients showed how COVID-19 pneumonia evolved from thesesmall nodules. The radiologic manifestation of the three cases will expand contemporary understanding of COVID-19.

Obesity and binge drinking often coexist and work synergistically to promote steatohepatitis; however, the underlying mechanisms remain obscure. In this mini-review, we briefly summarize clinical evidence of the synergistical effect of obesity and heavy drinking on steatohepatitis and discuss the underlying mechanisms obtained from the study of several mouse models. High-fat diet (HFD) feeding and binge ethanol synergistically induced steatohepatitis and fibrosis in mice with significant intrahepatic neutrophil infiltration; such HFD-plus-ethanol treatment markedly up-regulated the hepatic expression of many chemokines with the highest fold (approximately 30-fold) induction of chemokine (C-X-C motif) ligand 1 (Cxcl1), which contributes to hepatic neutrophil infiltration and liver injury. Furthermore, HFD feeding activated peroxisome proliferator-activated receptor gamma that subsequently inhibited CXCL1 upregulation in hepatocytes, thereby forming a negative feedback loop to prevent neutrophil overaction; whereas binge ethanol blocked this loop and then exacerbated CXCL1 elevation, neutrophil infiltration, and liver injury. Interestingly, inflamed mouse hepatocytes attracted neutrophils less effectively than inflamed human hepatocytes due to the lower induction of CXCL1 and the lack of the interleukin (IL)-8 gene in the mouse genome, which may be one of the reasons for difficulty in development of mouse models of alcoholic steatohepatitis and nonalcoholic steatohepatitis (NASH). Hepatic overexpression of Cxcl1 and/or IL-8 promoted steatosis-to-NASH progression in HFD-fed mice by inducing neutrophil infiltration, oxidative stress, hepatocyte death, fibrosis, and p38 mitogen-activated protein kinase activation. Collectively, obesity and binge drinking synergistically promote steatohepatitis via the induction of CXCL1 and subsequent hepatic neutrophil infiltration.

Objective To establish a quality control standard for Qingre Baidu granules .Methods Isatidis Radix ,Fruc-tus Forsythiae ,Herba Violae ,and Glycyrrhizae were identified by TLC ,and the concentration of chlorogenic acid was deter-minedbyHPLC.ThismethodwasemployedonanAgilentZORBAXSB-C18column(4.6mm×250mm,5μm)at30℃ witha mobile phase of acetonitrile (A) and 0 .2% formic acid (B) using the gradient elution program shown as follows :0-12 min , 11%-12% A run at the flow rate of 1 .0 ml/min .The injection volume was 20 μl and the detection wavelength was 327 nm . Results Characteristic spots could be detected by TLC and the specificity of the method was satisfactory .As for chlorogenic acid ,the equation of linear regression of chlorogenic acid was Y=60 .239 4X+9 .096 3 (r=0 .999 9) with the linear range of 6.19-396 .00 μg/ml .The average recovery was 99 .66% (RSD=2 .82% ) .Conclusion The established method is simple ,reli-able ,reproducible ,and can be used for the quantitative determination and quality control of Qingre Baidu granules .

The sigma-1 receptor is a molecular chaperone protein highly enriched in the brain. Recent studies linked it to many diseases, such as drug addition, Alzheimer's disease, stroke, depression, and even cancer. Sigma-1 receptor is enriched in lipid rafts, which are membrane microdomains essential in signaling processes. One of those signaling processes is ADAM17- and ADAM10-dependent ectodomain shedding. By using an alkaline phosphatase tagged substrate reporter system, we have shown that ADAM10-dependent BTC shedding was very sensitive to both membrane lipid component change and sigma-1 receptor agonist DHEAS treatment while ADAM17-dependent HB-EGF shedding was not; and overexpression of sigma-1 receptor diminished ADAM17- and ADAM10-dependent shedding. Our results indicate that sigma-1 receptor plays an important role in modifying the function of transmembrane proteases.
ADAM Proteins ; metabolism ; ADAM10 Protein ; ADAM17 Protein ; Amyloid Precursor Protein Secretases ; metabolism ; Animals ; Betacellulin ; COS Cells ; Cercopithecus aethiops ; Gene Expression ; HEK293 Cells ; Heparin-binding EGF-like Growth Factor ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Membrane Microdomains ; metabolism ; Membrane Proteins ; metabolism ; Receptors, sigma ; agonists ; metabolism