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ObjectiveTo study the effect and mechanism of Yixintai on mitochondrial fission proteins in the rat model of chronic heart failure. MethodTen of 60 SD rats were randomly selected as the sham operation group， and the remaining 50 rats were subjected to ligation of the left anterior descending coronary artery for the modeling of heart failure post myocardial infarction. The successfully modeled rats were randomized into model， low-， medium-， and high-dose （1.4， 2.8， and 5.6 g·kg-1， respectively） Yixintai， and trimetazidine （10 mg·kg-1） groups. The rats were administrated with corresponding doses of drugs by gavage， and the rats in the model group and sham operation group were given an equal volume of normal saline by gavage for 28 consecutive days. Enzyme-linked immunosorbent assay （ELISA） was then employed to measure the levels of amino-terminal pro-B-type natriuretic peptide （NT-pro BNP）， B-type natriuretic peptide （BNP）， and adenosine triphosphate （ATP） in the serum. Color Doppler ultrasound imaging was conducted to examine the cardiac function indicators. Hematoxylin-eosin staining and Masson staining were conducted to observe the pathological changes in the heart， and Image J was used to calculate collagen volume fraction （CVF）. Transmission electron microscopy was employed to observe the ultrastructural changes of myocardial cells. Terminal-deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to measure the apoptosis rate of myocardial cells. Western blot was employed to determine the protein levels of mitochondrial fission protein 1 （Fis1） and mitochondrial fission factor （Mff） in the outer mitochondrial membrane of the myocardial tissue. ResultCompared with the sham operation group， the model group showed elevated levels of NT-pro BNP and BNP in the serum， decreased ATP content， left ventricular ejection fraction （LVEF）， and left ventricular fraction shortening （LVFS）， increased left ventricular end-diastolic diameter （LVIDd） and left ventricular end-systolic diameter （LVIDs）， disarrangement of myocardial cells， inflammatory cell infiltration， increased collagen fibers and CVF， damaged myocardium and mitochondria， and increased apoptosis rate of myocardial cells， and up-regulated expression of Fis1 and Mff in the cardiac tissue （P<0.01）. Compared with the model group， different doses of Yixintai and trimetazidine lowered the serum levels of NT-pro BNP and BNP （P<0.05）， increased the ATP content （P<0.05）， increased LVEF and LVFS （P<0.01）， decreased LVIDd and LVIDs （P<0.01）. Moreover， the drugs alleviated the myocardial inflammatory damage and fibrosis， reduced CVF （P<0.01）， repaired the myocardial mitochondrial structure， and decreased the apoptosis rate of myocardial cells （P<0.01）. Medium- and high-dose Yixintai and trimetazidine down-regulated the expression of Fis1 and Mff in the myocardial tissue （P<0.05）. ConclusionYixintai can improve mitochondrial structure， reduce myocardial cell apoptosis， and improve cardiac function by inhibiting the expression of Fis1 and Mff in the myocardial tissue.
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ObjectiveTo study the medication rules of Professor. WANG Xingkuan and inherit his academic experience in the treatment of chest stuffiness and pain with the aid of the Traditional Chinese Medicine Inheritance Computing Platform V3.0 （TCMICS V3.0）. MethodThe original medical records of patients with angina pectoris in coronary heart disease （CHD） diagnosed and treated by Prof. WANG in the outpatient department of Hunan University of Chinese Medicine from 2017 to 2020 were collected and entered into the TCMICS V3.0. The rules of prescriptions and drugs were analyzed by the software. ResultA total of 1 044 prescriptions of Prof. WANG for the treatment of chest stuffiness and pain were collected. Most of the drugs were sweet and bitter in flavor and mainly acted on the lung meridian， followed by heart， spleen， liver， stomach， and kidney meridians. Among the prescriptions， Shengmaisan was the most commonly used classic prescription， and Xintongling No. Ⅲ was the top experienced prescription. High-frequency drugs mainly included Ophiopogonis Radix， Pinelliae Rhizoma， Salviae Miltiorrhizae Radix et Rhizoma， Trichosanthis Pericarpium， Coptidis Rhizoma， Schisandrae Chinensis Fructus， and Bupleuri Radix. The common doses of drugs were 3， 5， 10， and 15 g. The analysis of formulation rules revealed 129 combinations of common drugs， 58 combinations with confidence > 0.99， and the core drugs of common syndromes. Six core drug combinations were obtained by drug clustering. ConclusionProfessor WANG treats chest stuffiness and pain based on syndrome differentiation following the principles of benefiting Qi， nourishing Yin， eliminating phlegm， resolving stasis， soothing liver， and promoting bile secretion， reflecting his academic idea of "regulation of multiple organs and comprehensive treatment". The core prescriptions can be used for reference by clinical practitioners， but further clinical and experimental studies are still needed to verify their efficacy.
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Objective:To effectively express the receptor binding domain (RBD) of SARS-CoV-2 spike protein in Pichia pastoris and to evaluate its immunogenicity. Methods:The gene encoding the RBD protein was synthesized and cloned into the pPICZαA plasmid. After linearization, the plasmid was transferred and integrated into the genome of Pichia pastoris. The expressed RBD protein in culture supernatant was analyzed by Western blot and Biolayer interferometry. After screening, a single clone expressing the RBD protein was selected. The high-level expression of RBD protein was achieved by optimizing the fermentation process, including the salt concentration adjusting of the medium and induction condition optimization (pH, temperature and duration). The immunogenicity of the expressed RBD protein was evaluated in a mouse model. Results:A single clone with a high expression level of RBD protein was obtained and named RBD-X33. The expression level of RBD protein in the fermentation supernatant reached up to 240 mg/L after optimization of the induction condition (HBSM medium, pH=6.5±0.3, 22℃ and 120 h). In the mouse experiment, the recombinant RBD protein was formulated with Alum+ CpG dual adjuvant and injected into mice. The binding IgG antibody levels were up to 2.7×10 6 tested by ELISA and the neutralizing antibody levels were up to 726.8 tested by live virus neutralizing antibody assay (prototype). Conclusions:The RBD protein could be efficiently expressed in Pichia pastoris and induce stronger immune response in animals. This study suggested that the recombinant SARS-CoV-2 RBD protein expressed in Pichia pastoris could serve as a candidate antigen in the development of SARS-CoV-2 vaccine.
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ObjectiveTo explore the effect of Buyang Huanwutang （BYHWT） on platelet function and inflammatory cytokines in the rat model of acute blood stasis. MethodThe model of acute blood stasis was established with SD rats by ice water bath combined with injection of epinephrine. Rats were randomly assigned into four groups： normal group， model group， BYHWT （3.2 g·kg-1） group， and aspirin （60 mg·kg-1） group. The rats were injected with epinephrine hydrochloride on day 8 after 7 days of modeling. The macroscopic indexes of triditional Chinese medicine （TCM） syndrome including tongue manifestation and pulse manifestation were observed， while hemorheological indexes， blood coagulation， and platelet aggregation were detected. The serum levels of the inflammatory cytokine matrix metalloprotein-9 （MMP-9） and the adhesion factor intercellular adhesion molecule-1 （ICAM-1） and were determined by enzyme-linked immunosorbent assay （ELISA）. ResultThe pulse distention of rats in the model group was lower than that in the normal group （P<0.01）， while BYHWT improved the pulse distention of the rats with the syndrome of blood stasis （P<0.01）. In the model group， the tongue showed the characteristics of blood stasis syndrome， with dark purple veins at the tongue bottom and lower values of R， G， B on the tongue surface than those in the normal group （P<0.01）， which， however， can be recovered by BYHWT （P<0.01）. The blood viscosity at high， medium， and low shear stress and the plasma viscosity in the model group were higher than those in the normal group （P<0.01， P<0.05）. Compared with the model group， BYHWT restored the whole blood viscosity under high， medium and low shear stress and plasma viscosity （P<0.05，P<0.01）. The model group had shorter prothrombin time （PT）， shorter thrombin time （TT）， and higher fibrinogen （FIB） than the normal group （P<0.05， P<0.01）. BYHWT improved the TT and reduced the FIB in the rats with blood stasis syndrome （P<0.01）. The platelet aggregation rate induced by arachidonic acid （AA） and adenosine diphosphate （ADP） in the model group was higher than that in normal group （P<0.01） and BYHWT decreased the platelet aggregation rate of the rats with blood stasis syndrome （P<0.01）. The results of scanning electron microscopy showed that the model group exhibited excessive platelet activation， obvious pseudopodia， and increased aggregation of platelets compared with the normal group， while platelet activation and aggregation were rare in the BYHWT group. The serum levels of MMP-9 and ICAM-1 in the model group were higher than those in the normal group （P<0.01）， which were decreased in the BYHWT group （P<0.05， P<0.01）. ConclusionThe SD rats with the syndrome of acute blood stasis induced by ice water bath combined with injection of epinephrine demonstrate obvious changes in platelet function and morphology， inflammation， and abnormal cell adhesion. In the treatment of acute blood stasis in rats， BYHWT may reduce thrombosis and improve blood consistency and cohesion by mitigating inflammation， down-regulating cell adhesion factor overexpression， and improving platelet shape and function.
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Objective To observe the effects of the intercellular gap junction (GJIC) composed of connexin 43(Cx43) in mesenchymal stem cells (MSCs) from different sources and their signals on the biological behavior of multiple myeloma (MM) lateral population cells (SP cells), and to explore its possible mechanism. Methods Mesenchymal stem cells (MSCs) from different sources were isolated and cultured. SP cells of MM cell line RPMI 8266 were sorted by flow cytometry. RT-PCR and Western blot were used to detect the expression of Cx43 gene and protein in MSCs, RPMI 8266 and SP cells from different sources. The effects of MSCs from different sources on SP cell cycle, Cx43 protein expression, colony formation ability in vitro, stem cell related gene expression, cytokine secretion and drug resistance were observed. Results There was no significant difference in morphology and phenotype between MM-MSCs and ND-MSCs. Both MM-MSCs and RPMI 8266 cells expressed a higher level of Cx43. Co-culture with MM-MSCs induced more SP cells to enter G0 phase (P<0.001). The expressions of c-myc, Kif4 and Sox2 genes in SP cells were significantly up-regulated, while the expression of Oct-4 gene was down-regulated. After adding α-GA, c-myc, Kif4 and Sox2 were down-regulated in varying degrees, but there was no significant difference. The expression of Cx43 was up-regulated by (31.00±2)% and (39.00±2)%, respectively. The colony formation ability in vitro was up-regulated, and the addition of α-GA could partially inhibit this effect. A small amount of c-myc, Kif4, Sox2 and Oct-4 genes were expressed in RPMI 8266. These genes were significantly up-regulated in SP cell subpopulation. MM-MSCs secreted high levels of interleukin (IL)-6. After co-culture with SP cells, the expressions of IL-6, IL-10 and TGF-β in the supernatant of MM-MSCs were up-regulated (P=0.0072, P=0.037). bFGF and IL-17 had no significant change. After adding α-GA, the levels of IL-6, IL-10 and TGF-β in the supernatant decreased. MM cells were sensitive to bortezomib (BTZ) induced apoptosis, but SP cells were less sensitive. Co-culture with MM-MSCs significantly reduced BTZ-mediated apoptosis. The addition of α-GA partially restored the sensitivity of MM cells to bortezomib. Conclusion MM-MSCs and multiple myeloma SP cells up-regulate the expression of Cx43 protein, form more GJIC, and promote the proliferation and drug resistance of SP cells by changing the cytokine secretion profile of MSCs, which may be one of the reasons for the recurrence of MM.
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ObjectiveTo compare the feasibility of establishing the rat model of acute coronary syndrome with combined blood stasis and poison by lipopolysacharide （LPS） injection， ligation of coronary artery and different combinations of the two methods. MethodA total of 225 male SD rats were randomly divided into sham operation group， simple coronary artery ligation group， first injected LPS group ［LPS（5 mg·kg）injection 24 h before coronary artery ligation］ and follow injected LPS group ［LPS（5 mg·kg）injection 10 min after coronary artery ligation］. The indexes of each group were detected at 3， 24， 72 h after modeling， and the model was comprehensively evaluated. The general state and macroscopic evaluation indexes of traditional Chinese medicine （TCM） syndrome （tongue and pulse） of rats in each group were observed. ECG and echocardiography were used to evaluate cardiac function， and the myocardial ischemia and infarction areas were measured by Evans blue/2，3，5-triphenyltetrazolium chloride （TTC） staining. The content of creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， creatine kinase （CK）， and troponin T （cTnT） in serum as well as interleukin-1 β （IL-1β） and IL-6 changes were determined by biochemical method or enzyme-linked immunosorbent assay （ELISA）. Hematology analyzer was adopted to determine the white blood cell （WBC） count， and the four coagulation indexes， platelet aggregation rate， hemorheology and other coagulation evaluation indexes were also detected. The myocardial tissue was observed by hematoxylin-eosin（HE）staining. ResultAfter 3 h of modeling， compared with the conditions in sham operation group， the R， G and B values of tongue of rats （P<0.01）， pulse amplitude （P<0.01）， and cardiac function in simple coronary artery ligation group were decreased， and the color of hypoglossal veins became purple（P<0.01）. The content of CK， LDH， cTnT， IL-1β and IL-6 in serum（P<0.05）， myocardial infarction area（P<0.01）， and total number of WBCs （P<0.05）were increased. Compared with simple coronary artery ligation group， first injected LPS group and follow injected LPS group had increased hypoglossal veins， decreased R value of tongue and elevated cTnT content （P<0.01）， while follow injected LPS group had reduced B value of tongue， decreased cardiac output （CO）（P<0.05）， increased IL-1β content， and thinned left ventricular anterior walls at end-systole （LVAWs）（P<0.01）. After 24 h of modeling， compared with sham operation group， simple coronary artery ligation group presented significantly decreased R， G and B values of tongue， lengthened purplish dark hypoglossal veins （P<0.01）， reduced pulse amplitude（P<0.01） and cardiac function， enlarged myocardial infarction area（P<0.01）， increased whole blood viscosity， platelet aggregation rate， fibrinogen （FIB）， shortened prothrombin time （PT） and thrombin time （TT）（P<0.01）， and elevated total number of WBCs （P<0.01）and content of CK， LDH， cTnT and IL-6 in serum（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the pulse amplitude， R， G and B values of tongue （P<0.01）， and ejection fraction （EF） and fractional shortening （FS） scores （P<0.05）dropped， and hypoglossal veins were deepened and lengthened（P<0.05）， and cTnT content was increased（P<0.01）in first injected LPS group and follow injected LPS group. However， follow injected LPS group had thinned LVPWs， increased LDH content， platelet aggregation rate（P<0.05）， myocardial infarction area， and total number of WBC， level of IL-1β（P<0.05）， and shortened TT（P<0.01）. Additionally， 72 h after modeling， compared with sham operation group， simple coronary artery ligation group showed significantly reduced pulse amplitude， lowered R， G and B values of tongue， thickened and lengthened hypoglossal veins（P<0.01）， decreased cardiac function， and increased content of cTnT， FIB， whole blood viscosity（P<0.01），platelet aggregation rate， level of IL-6 and IL-1β（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the hypoglossal veins of the first injected LPS group and the follow injected LPS group were more purple， and the content of cTnT was boosted（P<0.01）， whereas follow injected LPS group had decreased pulse amplitude， R， G and B values of tongue， EF and FS scores （P<0.05）， and enlarged myocardial infarction area（P<0.01）. ConclusionCompared with the other modeling methods and models at different modeling time， the established model by LPS injection 10 min after coronary artery ligation for 24 h was more consistent with the clinical characteristics of acute coronary syndrome with combined blood stasis and poison.
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ObjectiveTo compare the feasibility of establishing the rat model of acute coronary syndrome with combined blood stasis and poison by lipopolysacharide （LPS） injection， ligation of coronary artery and different combinations of the two methods. MethodA total of 225 male SD rats were randomly divided into sham operation group， simple coronary artery ligation group， first injected LPS group ［LPS（5 mg·kg）injection 24 h before coronary artery ligation］ and follow injected LPS group ［LPS（5 mg·kg）injection 10 min after coronary artery ligation］. The indexes of each group were detected at 3， 24， 72 h after modeling， and the model was comprehensively evaluated. The general state and macroscopic evaluation indexes of traditional Chinese medicine （TCM） syndrome （tongue and pulse） of rats in each group were observed. ECG and echocardiography were used to evaluate cardiac function， and the myocardial ischemia and infarction areas were measured by Evans blue/2，3，5-triphenyltetrazolium chloride （TTC） staining. The content of creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， creatine kinase （CK）， and troponin T （cTnT） in serum as well as interleukin-1 β （IL-1β） and IL-6 changes were determined by biochemical method or enzyme-linked immunosorbent assay （ELISA）. Hematology analyzer was adopted to determine the white blood cell （WBC） count， and the four coagulation indexes， platelet aggregation rate， hemorheology and other coagulation evaluation indexes were also detected. The myocardial tissue was observed by hematoxylin-eosin（HE）staining. ResultAfter 3 h of modeling， compared with the conditions in sham operation group， the R， G and B values of tongue of rats （P<0.01）， pulse amplitude （P<0.01）， and cardiac function in simple coronary artery ligation group were decreased， and the color of hypoglossal veins became purple（P<0.01）. The content of CK， LDH， cTnT， IL-1β and IL-6 in serum（P<0.05）， myocardial infarction area（P<0.01）， and total number of WBCs （P<0.05）were increased. Compared with simple coronary artery ligation group， first injected LPS group and follow injected LPS group had increased hypoglossal veins， decreased R value of tongue and elevated cTnT content （P<0.01）， while follow injected LPS group had reduced B value of tongue， decreased cardiac output （CO）（P<0.05）， increased IL-1β content， and thinned left ventricular anterior walls at end-systole （LVAWs）（P<0.01）. After 24 h of modeling， compared with sham operation group， simple coronary artery ligation group presented significantly decreased R， G and B values of tongue， lengthened purplish dark hypoglossal veins （P<0.01）， reduced pulse amplitude（P<0.01） and cardiac function， enlarged myocardial infarction area（P<0.01）， increased whole blood viscosity， platelet aggregation rate， fibrinogen （FIB）， shortened prothrombin time （PT） and thrombin time （TT）（P<0.01）， and elevated total number of WBCs （P<0.01）and content of CK， LDH， cTnT and IL-6 in serum（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the pulse amplitude， R， G and B values of tongue （P<0.01）， and ejection fraction （EF） and fractional shortening （FS） scores （P<0.05）dropped， and hypoglossal veins were deepened and lengthened（P<0.05）， and cTnT content was increased（P<0.01）in first injected LPS group and follow injected LPS group. However， follow injected LPS group had thinned LVPWs， increased LDH content， platelet aggregation rate（P<0.05）， myocardial infarction area， and total number of WBC， level of IL-1β（P<0.05）， and shortened TT（P<0.01）. Additionally， 72 h after modeling， compared with sham operation group， simple coronary artery ligation group showed significantly reduced pulse amplitude， lowered R， G and B values of tongue， thickened and lengthened hypoglossal veins（P<0.01）， decreased cardiac function， and increased content of cTnT， FIB， whole blood viscosity（P<0.01），platelet aggregation rate， level of IL-6 and IL-1β（P<0.05）. Compared with the conditions in simple coronary artery ligation group， the hypoglossal veins of the first injected LPS group and the follow injected LPS group were more purple， and the content of cTnT was boosted（P<0.01）， whereas follow injected LPS group had decreased pulse amplitude， R， G and B values of tongue， EF and FS scores （P<0.05）， and enlarged myocardial infarction area（P<0.01）. ConclusionCompared with the other modeling methods and models at different modeling time， the established model by LPS injection 10 min after coronary artery ligation for 24 h was more consistent with the clinical characteristics of acute coronary syndrome with combined blood stasis and poison.
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Ulcerative colitis (UC) is a chronic and recurrent inflammatory bowel disease (IBD) that has become a major gastroenterologic problem during recent decades. Numerous complicating factors are involved in UC development such as oxidative stress, inflammation, and microbiota disorder. These factors exacerbate damage to the intestinal mucosal barrier. Spirulina platensis is a commercial alga with various biological activity that is widely used as a functional ingredient in food and beverage products. However, there have been few studies on the treatment of UC using S. platensis aqueous extracts (SP), and the underlying mechanism of action of SP against UC has not yet been elucidated. Herein, we aimed to investigate the modulatory effect of SP on microbiota disorders in UC mice and clarify the underlying mechanisms by which SP alleviates damage to the intestinal mucosal barrier. Dextran sulfate sodium (DSS) was used to establish a normal human colonic epithelial cell (NCM460) injury model and UC animal model. The mitochondrial membrane potential assay 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and staining with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) and Hoechst 33258 were carried out to determine the effects of SP on the NCM460 cell injury model. Moreover, hematoxylin and eosin (H&E) staining, transmission electron microscopy (TEM), enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qPCR), western blot, and 16S ribosomal DNA (rDNA) sequencing were used to explore the effects and underlying mechanisms of action of SP on UC in C57BL/6 mice. In vitro studies showed that SP alleviated DSS-induced NCM460 cell injury. SP also significantly reduced the excessive generation of intracellular reactive oxygen species (ROS) and prevented mitochondrial membrane potential reduction after DSS challenge. In vivo studies indicated that SP administration could alleviate the severity of DSS-induced colonic mucosal damage compared with the control group. Inhibition of inflammation and oxidative stress was associated with increases in the activity of antioxidant enzymes and the expression of tight junction proteins (TJs) post-SP treatment. SP improved gut microbiota disorder mainly by increasing antioxidant enzyme activity and the expression of TJs in the colon. Our findings demonstrate that the protective effect of SP against UC is based on its inhibition of pro-inflammatory cytokine overproduction, inhibition of DSS-induced ROS production, and enhanced expression of antioxidant enzymes and TJs in the colonic mucosal barrier.
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Animais , Camundongos , Antioxidantes/farmacologia , Colite/prevenção & controle , Colite Ulcerativa/metabolismo , Colo/metabolismo , Sulfato de Dextrana/toxicidade , Modelos Animais de Doenças , Microbioma Gastrointestinal , Inflamação/metabolismo , Camundongos Endogâmicos C57BL , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , SpirulinaRESUMO
Objective:To explore the mediating role of boredom tendency in anxiety, depression and meaning in life in college students.Methods:Totally 590 college students in Shijiazhuang were selected to carry out a questionnaire survey, including general questionnaire, boredom proneness questionnaire (BPQ), meaning in life questionnaire (MLQ), patient health questionnaire (PHQ-9) and general anxiety disorder-7(GAD-7). Spearman correlation analysis and hierarchical regression analysis were conducted to analyze the data by using SPSS 20.0, and Amos 24.0 was used to analyze the mediating effect of boredom tendency on anxiety, depression and sense of life meaning in college students.Results:Correlation analysis found that the total score of boredom tendency was negatively correlated with meaning in life ( r=-0.260, P<0.05) and education ( r=-0.110, P<0.05). The total score of boredom tendency was positively correlated with depression ( r=0.649, P<0.05) and anxiety ( r=0.555, P<0.05). The mediating effect model fit well(χ 2/ df=0.582, GFI=1.000, AGFI=0.994, RMSEA=0.000). Boredom tendency played a partial mediating role between anxiety meaning in life(standard effect size=-0.054), a complete mediating role between depression and meaning in life(standard effect size=-0.175), and a complete mediating role between educational background and meaning in life(standard effect size=0.025). Conclusion:Boredom tendency plays a mediating role in the model between related factors and meaning in life, and attentions should be paid to the psychological development of college students.
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Objective To construct 131 I-the fifth generation polyamidoamine (PAMAM(G5.0)) with targeting peptide Ser-Arg-Glu-Ser-Pro-His-Pro (SRESPHP;SR) or Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR;GP) and double targeting peptide SR/GP,and evaluate the targeting ability in medullary thyroid carcinoma (MTC) model.Methods PAMAM(GS.0),PAMAM(GS.0)-SR,PAMAM(GS.0)-GP and PAMAM(GS.0)-SR/GP were radiolabeled with 131I by chloramine T method.The radiolabeled yield and radiochemical purity were determined by thin layer chromatography.MTC xenografts were developed and the percentage radio-activity of injection dose per gram of tissue (％ID/g) in tumor and organs was measured at 24 h post-injection.Region of interest (ROI) was drawn and the tumor/non-tumor (T/NT) ratios at 4,8 and 24 h post-injection were calculated and compared among different groups.One-way analysis of variance,repetitive measurement analysis of variance and Dunnett-t test were used to compare the data of different groups.The relationship between ％ID/g and T/NT was analyzed with Pearson correlation.Results The radiolabeled yield was more than 75％ and radiochemistry purity was more than 90％.The difference of ％lD/g at 24 h post-injection was significant (F=14.400,P＜0.001) in tumors of all groups.The radioactive uptake in tumor of 131I-PAMAM (G5.0)-SR group was the highest at 24 h post-injection ((1.80± 0.18) ％ID/g).There were significant differences of T/NT ratios among different groups (F =4.776,P＜ 0.05)and between different time points (F =8.630,P＜0.05).Compared with negative control group (Na131 I),the T/NT ratios significantly increased in 131I-PAMAM(G5.0)-SR group at 4,8 and 24 h post-injection (t=4.169,7.123 and 4.032,all P＜0.05) and in 131I-PAMAM(G5.0)-GP group at 4 h post-injection (t =5.893,P＜0.05).The T/NT ratio in 131I-PAMAM (G5.0)-SR group was higher than that in 131 I-PAMAM (G5.0)-GP group at 24 h post-injection (t=2.871,P＜0.05).Conclusions PAMAM(G5.0)-SR,PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP can target the MTC models.131I-PAMAM(G5.0)-SR has the best biological properties and may provide a new precision method for MTC diagnosis,treatment and prognosis evaluation.
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Objective: To investigate the therapeutic effects of YiXuanNing Granule on the ischemic vertigo of the mice and rats, and to clarify its effects on the inflammation damage and oxidative stress in brain tissue of the mice and rats Methods: The vertigo models of mice and rats were established by mechanical vertigo and ligation of bilateral common carotid arteries (CCA), respectively. A total of 60 mice and 60 rats were selected and randomly divided into normal control group, model group, low dose of YiXuanNing Granule group (2.25 g middot; kg-1/1. 50 g middot; kg-1), middle dose of YiXuanNing Granule group (4.50 g middot; kg-1/3. 00 g middot; kg-1), high dose of YiXuanNing Granule group (9.00 g middot; kg-1/6. 00 g middot; kg-1) and positive control group (1.50 g middot; kg-1/1. 00 mg middot; kg -1 flunarizine hydrochloride capsule), and there were 10 animals in each group. The mice and the rats in control group and model group were intragastrically administered with 10 mL middot; kg-1 normal saline, and the mice and the rats in the other groups were given 10 mL middot; kg: corresponding drug once a day for 7 d. The escape time of jumping off the platform of the mice and the rats in various groups were determined; the food intake of the mice in various groups were detected; the activities of superoxide dismutase (SOD) and the levels of malondialdehyde (MDA) in brain tissue of the rats in various groups were detected by colorimetric method. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in brain tissue of the rats in various groups were detected by ELISA method. Results: Compared with model group, the escape time of jumping off the platform of the mice and the rats in different doses of YiXuanNing Granule groups and positive control group was shortened (P<0. 05 or P<0. 01), and the food intake of the mice in various groups was increased. Compared with model group, the SOD activities in brain tissue of the rats in different doses of YiXuanNing Granule groups were significantly increased (P<0. 05 or P<0. 01); the MDA levels in brain tissue of the rats in different doses of YiXuanNing Granule groups were significantly decreased (P<0. 01); compared with model group, the levels of TNF-α and IL-1β in brain tissue of the rats and the mice in middle and high doses of YiXuanNing Granule groups were significantly decreased (P<0. 05 or P<0. 01). Conclusion: YiXuanNing Granule can inhibit the inflammatory damage in brain tissue of the rats with ischemic vertigo and reduce the oxidative stress, thereby, it can play a therapeutic role in ischemic vertigo.
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Objective@#To construct 131I-the fifth generation polyamidoamine (PAMAM(G5.0)) with targeting peptide Ser-Arg-Glu-Ser-Pro-His-Pro (SRESPHP; SR) or Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR; GP) and double targeting peptide SR/GP, and evaluate the targeting ability in medullary thyroid carcinoma (MTC) model.@*Methods@#PAMAM(G5.0), PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP were radiolabeled with 131I by chloramine T method. The radiolabeled yield and radiochemical purity were determined by thin layer chromatography. MTC xenografts were developed and the percentage radio-activity of injection dose per gram of tissue (%ID/g) in tumor and organs was measured at 24 h post-injection. Region of interest (ROI) was drawn and the tumor/non-tumor (T/NT) ratios at 4, 8 and 24 h post-injection were calculated and compared among different groups. One-way analysis of variance, repetitive measurement analysis of variance and Dunnett-t test were used to compare the data of different groups. The relationship between %ID/g and T/NT was analyzed with Pearson correlation.@*Results@#The radiolabeled yield was more than 75% and radiochemistry purity was more than 90%. The difference of %ID/g at 24 h post-injection was significant (F=14.400, P<0.001) in tumors of all groups. The radioactive uptake in tumor of 131I-PAMAM(G5.0)-SR group was the highest at 24 h post-injection((1.80±0.18) %ID/g). There were significant differences of T/NT ratios among different groups(F=4.776, P<0.05)and between different time points(F=8.630, P<0.05). Compared with negative control group (Na131I), the T/NT ratios significantly increased in 131I-PAMAM(G5.0)-SR group at 4, 8 and 24 h post-injection (t=4.169, 7.123 and 4.032, all P<0.05) and in 131I-PAMAM(G5.0)-GP group at 4 h post-injection (t=5.893, P<0.05). The T/NT ratio in 131I-PAMAM(G5.0)-SR group was higher than that in 131I-PAMAM(G5.0)-GP group at 24 h post-injection (t=2.871, P<0.05).@*Conclusions@#PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP can target the MTC models. 131I-PAMAM(G5.0)-SR has the best biological properties and may provide a new precision method for MTC diagnosis, treatment and prognosis evaluation.
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Objective To discuss function of the fusion cells of human bone marrow stromal cells (BMSCs) and human myeloma cell RPMI 8226 in the pathogenesis of multiple myeloma bone disease.Methods The cells,labeled by cell tracer green fluorescent probe (CMFDA) and red fluorescent probe (CMTMR),respectively,were induced into fusion by chemical polyethylene glycol (polyethyleneglycol,PEG-1000),and cell fusion model was set up.Whether fusion cells had nucleus fusion was determined by Karyotype analysis.The expressions of stemness-related genes,SIRP α gene and DC-STAMP gene in fusion cells were identified.Results Polyethylene glycol (PEG-1000) could mediate the integration of BMSCs and RPMI 8226 cells.The number of chromosomes in more than 80 ％ the hybrid cells was about 80.Fusion cells not only showed that BMSCs,stemness-related genes of c-myc,Klf-4 and OCT-4 genes expressed positively,but also the fusion-related genes SIRPα and DC-STAMP expressed positively.Conclusion BMSCs and RPMI 8226 cells can form fusion cells,and the cells have the potential for further integration,which is one of the important reasons for the promotion of muhiple myeloma bone destruction.
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Objective To detect and separate the side population cells(SP) from multiple myeloma (MM) cell lines PRIM8226,and to study their biological characteristics.Methods Fluorescence activated cell sorter (FACS) and Hoechst33342/PI dye were used to sort SP cells of PRIM8226.The multiplication capacity was tested by the growth curve and MTT test,SP cells proportion and the cell cycle were analyzed by flow cytometry,the colony-formtion ability was compared in terms of colony forming experiment,the expression of c-myc,KIF4,SOX2,OCT4 was tested by RT-PCR.The oncogenicity of the cells was analyzed by nude mouse transplantation tumor experiment in vivo.Results The ratio of SP cells in PRIM8226 was (1.78±0.89) ％.More SP cells in the G0 / G1 period,(44.34±3.09) ％ vs (28.49±1.97) ％,P ＜ 0.05,and fewer cells in S phase than MP cells,(38.83±3.69) ％ vs (51.49±4.62) ％,P ＜ 0.05.There were no difference in the expression of CD38 and CD138 between SP cells and MP cells,respectively,(78.5±8.5) ％ vs (82.0±4.0) ％ and (72.3±15.7) ％ vs (84.3±11.9) ％,P ＞ 0.05.Colony formation assay showed that the colony forming efficiency of the SP cells was higher than the MP cells,single cell clone diameter,the number of clone forming,the clone formtion rate were significantly higher than MP cells,0.280±0.016 vs 0.118±0.019,1 722±127 vs 358±14,(86.1±3.46) ％ vs (17.9±1.88) ％,P ＜ 0.05.The mRNA expression levels of c-myc,KIF4,SOX2,OCT4 in SP cells were higher than those in MP cells,c-myc (29.90±3.73) ％ vs (16.84±2.35) ％,KIF4 (29.97±2.89) ％ vs (19.06±1.23) ％,SOX2 (40.00±4.58) ％ vs (16.62±2.09) ％,OCT4 (32.96±0.92) ％ vs (23.27±0.92) ％,all P ＜ 0.05.Nude mouse transplantation tumor experiment in vivo showed the oncogenicity of the SP cells was more stronger than MP cells (5×103 vs 5×l05).Conclusion There are notably difference in the cell cycle,colony formation assay,the mRNA expression levels and oncogenicity,but no difference in the expression of CD38 and CD138,the proliferation ability between SP cells and MP cells.
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Objective To investigate the exposure and protection of recurrent laryngeal nerve (RLN) in the reoperation for thyroid diseases.Methods Clinical data of 214 cases undergoing thyroid reoperation were retrospectively analyzed.The patients with a short interval between the 2 thyroid operations or with external-infiltrated thyroid cancer were approached at the lateral strap muscles and the leading edge of the sternocleidomastoid.RLNs were exposed in the lateral region of superior mediastinum tracheoesophageal groove or at the point where RLN enters to throat.RLNs of patients with lymph node metastasis were exposed beside the enlarged lymph nodes.The patients with a long interval between the 2 thyroid operations and with benign tumor or tumor without external infiltration were exposed their thyroids at the anterior midline and then RLNs were exposed at the posterior lateral of the middle thyroid veins or at the inferior thyroid artery.Results Among the 214 cases,344 RLNs were anatomically exposed including 188 right and 156 left.84 cases had single exposure and 130 cases had bilateral exposure.44 RLNs were exposed at the point where RLN enters to throat,104 RLNs at the posterior lateral of the middle thyroid veins,40 RLNs at the inferior thyroid artery,124 RLNs at the lateral region of superior mediastinum tracheoesophageal groove,and 32 RLNs beside the enlarged lymph nodes.For the 2 cases suffering hoarse voice the day after they underwent thyroid operation in other hospital,suture ligation at the the entrance point was found when they received the reoperation in our hospital.Three of the total 344 RLNs (0.87％ ) had RLN branch injury in the entire group.Conclusion It is possible to reduce RLN injury during the reoperation for thyroid disease if surgeons are familiar with the dissection of RLN under normal or pathological condition,avoid adhesive or scar tissues,and select the appropriate anatomic approach.
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Objective To explore the rule of central region lymph nodes metastasis in differentiated thyroid carcinoma (DTC) and the significance of central region lymph nodes neck dissection in surgical operation for DTC.Methods The clinical and pathological data of 122 lymph nodes in 109 patients (31 female and 78 male) with DTC undergoing neck lymph nodes ( central region lymph nodes included) dissection from Jan.2003 to Jun.2007 in our hospital were analyzed retrospectively.According to preoperative physical examination and imaging analysis,patients were divided into clinical cervical lymph nodes metastasis ( cN + ) group and clinical no cervical lymph nodes metastasis (cNO) group and compared respectively with their pathological data after surgery.Resuits Of the 122 lymph nodes,lymph node metastasis rate was significantly higher in region Ⅵ than in region Ⅱ,Ⅲ and Ⅳ.The difference had statistical significance (P ＜0.01 ).65.6％ (80/122) metastasis was in the central region.81.2％ (56/69) patients in cN + group and 45.3％ (24/53) patients in cN0 group had central region lymph nodes metastasis.Conclusions Central region lymph nodes metastasis is common for patients with DTC.Routine neck dissection in central region should be done in DTC operation.
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In the present study, the regulation of Vitreoscilla hemoglobin (VHb) on astragaloside IV biosynthesis was investigated. An intermediate expression vector consisting of the CaMV35S promoter fused to the vgb and nopaline synthase terminator was transferred into Astragalus membranaceus via Agrobacterium rhizogenes. The transgenic hairy roots were confirmed by PCR amplification and Southern blot hybridization. The expression of vgb in transgenic hairy roots was confirmed by RT-PCR. After 15 days cultivation, the dry weight and growth rate of transgenic hairy roots were higher than that of the non-transgenic hairy root. ELSD-HPLC analysis showed that astragaloside IV content of transgenic hairy roots was 5 to 6 times of non-transgenic hairy root control and 10 to 12 times of Radix Astragali from Shanxi Province. These results suggested that the expression of vgb promoted the growth of transgenic hairy roots, and increased the content of astragaloside IV.
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Objective To discuss the prevention and treatment of the complications of modified radical mastectomy with nipple-areola complex (NAC) sparing. Methods The clinical data of 124 breast cancer patients who had a NAC sparing modified radical mastectomy from January 1998 to December 2008 were analyzed retrospectively. The causes of complications were evaluated and the proposal of prevention and treatment was put forward. Results Of the 124 patients, 18 had nipple necrosis (14.52%), 9 had skin infections and necrosis (7.26%), 7 subcutaneous hydrops(5.65%), 5 upper extremity lymphedema (4.03%), 3 chest muscle contracture with ipsilateral upper limb dyskinesia (2.42%), and 2 operative residual cavity hemorrhage(1.61%). Conclusion All the post-operative complications listed above could be possibly avoided by more careful operation procedure.
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OBJECTIVE: To establish a supplementary diagnostic indicator (infrared radiant intensity) in tongue diagnosis of traditional Chinese medicine (TCM) in patients with hyperplasia of mammary glands through correlation analysis of infrared radiant intensity between hyperplastic breast tissue and tongue surface. METHODS: Infrared radiant intensity of the hyperplastic breast tissue and different points on tongue surface in 20 cases of hyperplasia of mammary glands with liver-energy stagnation and phlegm retention syndrome and 16 cases of hyperplasia of mammary glands with irregular thoroughfare and conception vessels syndrome were measured with external infrared spectrometer PHE-201 made by Shanghai Institute of Technical Physics. Correlation of infrared radiant intensity between the hyperplastic breast tissue and the different points on tongue surface was assessed by using bivariate correlation analysis. RESULTS: The results showed that the numbers of positive correlated wave bands of infrared radiant intensity between the hyperplastic breast tissue and different detected points on tongue surface in the patients with liver-energy stagnation and phlegm retention syndrome and irregular thoroughfare and conception vessels syndrome were 127 (83.55%) and 71 (46.71%), respectively. Infrared radiant intensity between the hyperplastic breast tissue and the tongue surface had a positive correlation. CONCLUSION: Infrared radiant intensity can be used as one of supplementary diagnostic indicators in TCM tongue diagnosis of hyperplasia of mammary glands.