This study aims to investigate the protective effect of naringenin(Nar)on cadmium(Cd)-induced liver injury in chickens and its regulatory role in endoplasmic reticulum stress(ERS)and autophagy.Forty-eight 1-day-old Hyland Brown egg roosters were divided into four groups:The control group,the Cd group(150 mg/kg CdCl2),the Nar group(250 mg/kg Nar),and the Cd+Nar group(150 mg/kg CdCl2+250 mg/kg Nar),and the rearing period was 60 days.On the 60th day,6 chickens were randomly selected from each group,and after fasting for 24 hours,blood was collected via vein and then removed from the neck and executed,the livers were collected and organ coefficients were calculated,changes in the content of liver function-related indexes were de-termined,pathological sections were made and observed,and the expression of mRNA and protein of related genes were detected by fluorescence quantitative PCR and Western blot method.The re-sults showed that Cd treatment resulted in a highly significant decrease(P＜0.01)in body mass,a highly significant increase(P＜0.01)in liver coefficient,a highly significant increase(P＜0.01)in serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and lactate dehydrogenase(LDH)levels were highly significantly increased(P＜0.01),hepatic antioxidant capacity was highly significantly reduced(P＜0.01),and ERS-related genes(GRP-78,PERK,eIF-2α,CHOP,ATF-4,ATF-6)and autophagy-related genes(Atg-5,Beclin-1,LC3)mRNA expression was highly significantly up-regulated(P＜0.01),and the expression of mRNA for p62 was highly significantly reduced(P＜0.01).After the addition of Nar,Cd-induced liver injury was significantly alleviated,liver function indexes improved,hepatic antioxidant capacity was highly significantly el-evated(P＜0.01),mRNA expression of ERS-related genes(GRP-78,PERK,eIF-2α,CHOP,ATF-4,ATF-6)and autophagy-related genes(Atg-5,Beclin-1,LC3)was highly significantly decreased(P＜0.01)and mRNA expression of P62 was highly significantly increased(P＜0.01).This study suggests that Nar has potential in alleviating Cd-induced liver damage in chickens through the reg-ulation of ERS and autophagy,providing theoretical support for its application in livestock health.

This study aims to investigate the protective effect of naringenin(Nar)on cadmium(Cd)-induced liver injury in chickens and its regulatory role in endoplasmic reticulum stress(ERS)and autophagy.Forty-eight 1-day-old Hyland Brown egg roosters were divided into four groups:The control group,the Cd group(150 mg/kg CdCl2),the Nar group(250 mg/kg Nar),and the Cd+Nar group(150 mg/kg CdCl2+250 mg/kg Nar),and the rearing period was 60 days.On the 60th day,6 chickens were randomly selected from each group,and after fasting for 24 hours,blood was collected via vein and then removed from the neck and executed,the livers were collected and organ coefficients were calculated,changes in the content of liver function-related indexes were de-termined,pathological sections were made and observed,and the expression of mRNA and protein of related genes were detected by fluorescence quantitative PCR and Western blot method.The re-sults showed that Cd treatment resulted in a highly significant decrease(P＜0.01)in body mass,a highly significant increase(P＜0.01)in liver coefficient,a highly significant increase(P＜0.01)in serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and lactate dehydrogenase(LDH)levels were highly significantly increased(P＜0.01),hepatic antioxidant capacity was highly significantly reduced(P＜0.01),and ERS-related genes(GRP-78,PERK,eIF-2α,CHOP,ATF-4,ATF-6)and autophagy-related genes(Atg-5,Beclin-1,LC3)mRNA expression was highly significantly up-regulated(P＜0.01),and the expression of mRNA for p62 was highly significantly reduced(P＜0.01).After the addition of Nar,Cd-induced liver injury was significantly alleviated,liver function indexes improved,hepatic antioxidant capacity was highly significantly el-evated(P＜0.01),mRNA expression of ERS-related genes(GRP-78,PERK,eIF-2α,CHOP,ATF-4,ATF-6)and autophagy-related genes(Atg-5,Beclin-1,LC3)was highly significantly decreased(P＜0.01)and mRNA expression of P62 was highly significantly increased(P＜0.01).This study suggests that Nar has potential in alleviating Cd-induced liver damage in chickens through the reg-ulation of ERS and autophagy,providing theoretical support for its application in livestock health.

ObjectiveTo explore the efficacy and mechanism of Danshen Zexie decoction in treating the rats with metabolic-associated fatty liver disease （MAFLD）. MethodSixty male SD rats were randomized into control group， model group， essentiale （polyene phosphatidylcholine capsules， 0.144 g·kg^-1） group， and low-， medium-， and high-dose Danshen Zexie decoction （1.16， 2.32， and 4.64 g·kg^-1， respectively） groups. The rat model of MAFLD was established with high fat diet， and 8-week therapy started at the induction of MAFLD. The serum levels of total cholesterol （TC）， triglyceride （TG）， alanine aminotransferase （ALT）， and aspartate aminotransferase （AST）， as well as the liver levels of TC， TG， free fatty acid （FFA）， superoxide dismutase （SOD）， glutathione （GSH）， and malondialdehyde （MDA）， were measured. The pathological changes of liver were observed by hematoxylin-eosin （HE） staining and oil red O staining， and the steatosis score and oil red O staining area were calculated. The level of reactive oxygen species （ROS） in the liver was detected by immunofluorescence. The protein levels of adenosine monophosphate-activated protein kinase （AMPK）， p-AMPK， nuclear factor E₂-related factor 2 （Nrf2）， glutamate-cysteine ligase catalytic subunit （GCLC）， glutamate-cysteine ligase modifier subunit （GCLM）， and nicotinamide adenine dinucleotide phosphate： quinine oxidoreductase （NQO1） in the liver were determined by Western blot. ResultCompared with control group， the modeling of MAFLD elevated the levels of TC， TG， ALT， and AST in the serum and TC， TG， FFA， MDA， and ROS in the liver （P<0.01）， lowered the levels of SOD and GSH and down-regulated the protein levels of phosphorylation（p）-AMPK， Nrf2， GCLC， GCLM， and NQO1 in the liver （P<0.05， P<0.01）. Further， a large number of lipid droplet vacuoles and balloon-like lesions appeared in the hepatocytes， and the steatosis score and oil red O staining area increased in the model group （P<0.01）. Compared with model group， Danshen Zexie decoction lowered the levels of TC， TG， ALT， and AST in the serum and TC， TG， FFA， MDA， and ROS in the liver （P<0.01）， increased the levels of SOD and GSH and up-regulated the protein levels of p-AMPK， Nrf2， GCLC， GCLM， and NQO1 in the liver （P<0.05， P<0.01）. Moreover， the decoction alleviated the degree of liver steatosis （P<0.05， P<0.01）. ConclusionDanshen Zexie decoction protects against MAFLD by activating AMPK/Nrf2 signaling pathway and suppressing oxidative stress.

Objective:To determine the expression of silent information regulator 1 (Sirt1) , Sirt3 and hypoxia-inducible factor 1α (HIF-1α) in cutaneous squamous cell carcinoma (CSCC) tissues and cells, and to explore their role in the occurrence and development of CSCC.Methods:From January 2019 to December 2020, 30 lesional skin tissues were obtained from patients with histopathologically confirmed poorly-, moderately- or well-differentiated CSCC, and 30 normal skin tissues were obtained from patients with non-cancerous diseases in Department of Dermatology, General Hospital of Ningxia Medical University. A CSCC cell line A431 and a human keratinocyte cell line HaCaT were cultured. Immunohistochemical study, Western blot analysis and real-time quantitative PCR (RT-PCR) were performed to determine the protein and mRNA expression of Sirt1, Sirt3 and HIF-1α in CSCC tissues of different grades of differentiation and normal skin tissues, cytochemical and immunofluorescence staining and RT-PCR were conducted to determine the protein and mRNA expression of Sirt1, Sirt3 and HIF-1α in A431 and HaCaT cells, respectively. Comparisons of measurement data among multiple groups were performed by using one-way analysis of variance, and comparisons between two groups by using t test. Results:Immunohistochemical study showed that the expression level of Sirt3 (expressed as the average optical density) was 100 ± 12.12, 117.72 ± 26.23, 127.32 ± 24.45, 132.71 ± 31.61 in the normal skin tissues and well-, moderately- and poorly-differentiated CSCC tissues respectively, and there was a significant difference among these groups ( F = 20.14, P < 0.001) ; the expression of Sirt1 and HIF-1α increased in turn from the normal skin tissues to the well-, moderately- and poorly-differentiated CSCC tissues, and significantly differred in these groups ( F = 174.50, 225.00, respectively, both P < 0.001) . As Western blot analysis revealed, the expression level of Sirt3 significantly differed among the normal skin tissues, well-, moderately- and poorly-differentiated CSCC tissues (expressed as relative gray value: 1.000 ± 0.132, 1.403 ± 0.411, 1.387 ± 0.393, 1.677 ± 0.683, respectively; F = 34.97, P < 0.001) , and so did the expression levels of Sirt1 and HIF-1α ( F = 69.29, 199.90, respectively, both P < 0.00l) , with a gradually increasing trend in their expression levels from the the normal skin tissues to well-, moderately- and poorly-differentiated CSCC tissues. RT-PCR showed that the mRNA expression of Sirt3, Sirt1 and HIF-1α was sequentially increased from the normal skin tissues to well-, moderately- and poorly-differentiated CSCC tissues, and significant differences were observed among these groups ( F = 113.00, 174.50, 50.33, respectively, all P < 0.001) . The protein expression levels of Sirt3, Sirt1 and HIF-1α were significantly higher in the A431 cells than in the HaCaT cells ( t = 16.75, 18.34, 27.76, respectively, all P < 0.001) , and so were their mRNA expression levels ( t= 14.22, 9.62, 16.86, respectively, all P < 0.001) . Conclusion:Increased expression of Sirt3, Sirt1 and HIF-1α was observed in CSCC tissues and cells, which may promote the occurrence and development of CSCC.

With the increasing cost of drug development and clinical trials, it is of great value to make full use of all kinds of data to improve the efficiency of drug development and to provide valid information for medication guidelines. Model-based meta-analysis (MBMA) combines mathematical models with meta-analysis to integrate information from multiple sources (preclinical and clinical data, etc.) and multiple dimensions (targets/mechanisms, pharmacokinetics/pharmacodynamics, diseases/indications, populations, regimens, biomarkers/efficacy/safety, etc.), which not only provides decision-making for all key points of drug development, but also provides effective information for rational drug use and cost-effectiveness analysis. The classical meta-analysis requires high homogeneity of the data, while MBMA can combine and analyze the heterogeneous data of different doses, different time courses, and different populations through modeling, so as to quantify the dose-effect relationship, time-effect relationship, and the relevant impact factors, and thus the efficacy or safety features at the level of dose, time and covariable that have not been involved in previous studies. Although the modeling and simulation methods of MBMA are similar to population pharmacokinetics/pharmacodynamics (Pop PK/PD), compared with Pop PK/PD, the advantage of MBMA is that it can make full use of literature data, which not only improves the strength of evidence, but also can answer the questions that have not been proved or can not be answered by a single study. At present, MBMA has become one of the important methods in the strategy of model-informed drug development (MIDD). This paper will focus on the application value, data analysis plan, data acquisition and processing, data analysis and reporting of MBMA, in order to provide reference for the application of MBMA in drug development and clinical practice.

OBJECTIVETo observe the difference in the clinical therapeutic effects on bronchial asthma in children of different body constitutions treated withplaster andplaster.

METHODSOne hundred and twenty-two children of bronchial asthma were divided into three groups according to TCM body constitutions, 42 cases in thedeficiency constitution group, 40 cases in thedeficiency constitution group and 40 cases in the phlegm damp constitution group. From 2011 to 2013, the acupoint plaster was applied to all of the children in the three groups during the dog days and the third nine-day period after the winter solstice each year. The average attack frequency and onset days of bronchial asthma and relevant immune function indicators were observed during treatment and 1 year after treatment in the children and the therapeutic effects were evaluated.

RESULTS①In 2014, the acute attacks of bronchial asthma were (1.2±0.9) times and (1.4±0.4) times in thedeficiency constitution group and thedeficiency constitution group, all lower than (3.0±0.5) times in the phlegm damp constitution group (both<0.05) separately. ②After treatment, in thedeficiency constitution group anddeficiency constitution group, the values of IgG, IgA and IgM were all increased as compared with those before treatment (all<0.05). ③The total effective rate was over 95% in the children of the three groups. The clinical control rates in thedeficiency constitution group and thedeficiency constitution group were higher apparently than that in the phlegm damp constitution group, indicating the significant difference statistically (both<0.05).

CONCLUSIONSThe combined treatment ofplaster andplaster are effective on bronchial asthma in the children of different body constitutions. The therapeutic effects for thedeficiency constitution and thedeficiency constitution are more apparent than that for the phlegm damp constitution.

Objective To explore the best keV conditions of show hepatocellular carcinoma lesions with the best contrast-to-noise ratio (CNR)and image quality,and evaluate the effect of monochromatic energy imaging on the detection of hepatocellular carcinomas.Methods The dual-energy CT imaging of 30 patients with hepatocellular carcinoma were analyzed retrospectively.Monochromatic energy images (40 to 140 keV,interval 10 keV)were produced by the dual-engery software.CNR and image noise of monochromatic energy,100 keV and 140 keV images were measured respectively.The CNR and the imaging noise were optimized.The results between the monochromatic energy images and the mixed energy images were compared by using the two related samples of non-parameter rank test.Results In the arterial phase, the monochromatic energy level with the highest CNR was concentrated at 70 keV,and the lowest level of the image noises was at 70 and 80 keV.The images at 70 keV were chosen to be compared with the fusion images.The CNR values of the monochromatic images at 70 keV were similar as those at 100 keV (1 .68 ± 1 .04 vs 1 .88 ± 1 .59 ,P =0 .149 ),and significantly higher than those at 140 keV (1 .68 ± 1 .04 vs 0.62±0.92,P =0.000).The image noises at 70 keV were similar as those at 140 keV (6.52±1.53 vs 7.60±2.73,P =0.050), and significantly lower than those at 100 keV (6.52±1.53 vs 8.55±1.11,P =0.000).Conclusion The monochromatic energy imaging at 70 keV can improve the CNR of the hepatocellular carcinoma lesions without reducing the image quality,which will help the lesion detection.

The male patient was 72 years old with 10-years hoarseness. Electronic laryngoscopy showed a great pink lobulated lesion at the left back of soft palate and lateral pharyngeal wall, close to the choanal region. Neck CT revealed irregular signal intensities of soft tissue at the left wall of nasopharynx, side wall and posterior wall of oropharynx and hypopharynx, bilateral submandibular and glottic region. The patient underwent operation treatment. Histopathology showed the mass was composed of identical mature rhabdomyoma cells, with scarce interstitial substance. The tumor cells was round, oval or polygonal with eosinophilic cytoplasmic granules, longitudinal grain or grain was occasionally found. Some tumor cells arranged in disorder with match structure. Pathological diagnosis was rhabdomyoma.
Aged ; Humans ; Male ; Pharynx ; pathology ; Rhabdomyoma ; diagnosis ; pathology ; Tomography, X-Ray Computed

To prepare recombinant human retinol binding protein 4 (RBP4) by using the baculovirus expression system and to detect its immunogenicity, the fusion DNA fragment of secretory signal peptide SS64 and human RBP4 gene was subcloned into a baculovirus transfer vector pFastBac-dual(pFBd), and the corresponding recombinant transfer plasmid was transformed into E. coli strain DH10bac, after transposition recombinant shuttle bacmid was screened out. The logarithmic phase Sf9 cells were transfected with the recombinant bacmid and then the recombinant baculovirus containing hRBP4 expression box were generated. After amplification of recombinant baculovirus, the recombinant baculovirus seeds were obtained. To express human RBP4, logarithmic phase Sf9 cells were infected with the virus seeds and SDS-PAGE and Western blotting were used to detect and identify the expression. Finally, to prepare a batch of RBP4 protein, logarithmic phase Sf9 cells in suspension culture were infected with recombinant baculovirus seeds and the supernatant was harvested after 120 hours post-infection for purification. Finally for preparation of polyclonal antibody and evaluation of immunogenicity, the recombinant hRBP4 from insect cells and from E. coli were immunized rabbits. Restriction enzyme digestion and sequencing confirmed that the recombinant baculovirus transfer plasmid was constructed correctly, and subsequently recombinant RBP4-bacmid was generated successfully. SDS-PAGE and Western blotting analysis suggested that human RBP4 protein was highly expressed in Sf9 cells with the molecular weight of approximately 23 kDa. The recombinant RBP4 protein could be secreted into the medium efficiently, and the expression level was calculated amount of 100 mg/L. Finally the rabbit antiserum was harvested after recombinant RBP4 immunization, therein the titer of antiserum against baculovirus recombinant RBP4 is 1:100 000 whereas the titer of antiserum against E. coli recombinant RBP4 is only 1:10 000. Overall, human RBP4 was high efficiently expressed successfully with good antigenicity in baculovirus system, and high affinity antiserum was obtained. A solid foundation was laid for the next step of the preparation of human serum RBP4 detection kit.
Animals ; Baculoviridae ; genetics ; Blotting, Western ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genetic Vectors ; Humans ; Immune Sera ; Insecta ; Rabbits ; Recombinant Proteins ; biosynthesis ; immunology ; Retinol-Binding Proteins, Plasma ; biosynthesis ; immunology ; Sf9 Cells ; metabolism ; Transfection

ObjectiveTo analyze the application value of diffusion-weighted magnetic resonance imaging (DWMRI) in gross tumor volume (GTV) delineation of esophageal squamous cell carcinoma (SCC).MethodsTwenty-nine patients with esophageal SCC treated with radical surgery were analyzed.Routine CT scan,MRI T2-weighted and DWMRI were employed before surgery;diffusion-sensitive gradient b-values were taken 400,600 and 800 s/mm2.GTVs were delineated using CT,MRI T2-weighted images and DWMRI under different b-value images.The length of GTVs measured under different images was compared with the pathological length and confirm the most accurate imaging condition.Use radiotherapy planning system to fuse DWMRI images and CT images to investigate the possibility of delineate GTVs on fused images.ResultsThe difference of GTV length value between CT,T2 WI images and specimen was 3.36 mm and 2.84 mm.When b =400,600 and 800 s/mm2,the difference between GTV length value on the DWMRI images and on specimen was 0.47 mm,-0.47 mm and - 1.53 mm;the correlation coefficient of the measuring esophageal lengths on DWMRI images and the pathological lengths was 0.928,0.927 and 0.938.DWMRI images and CT images could fuse accurately on radiotherapy planning system.GTV margin could.show clearly on fused images.ConclusionsDWMRI images can display the esophageal carcinoma lengths and margin accurately.When DWMRI images fused with CT images,GTV margin could show clearly,it can be used to delineate GTV accurately.