Aim To explore the effects of Shuangshen tongguan(SSTG) drug serum on Calcium-Calmodulin(CaM)-Calcium/CaM dependent kinaseⅡsignal system in cardiomyocytes injury induced by hypoxia and reoxgyenization.Methods The cardiomyocytes were deprived of oxygen and glucose to mimic hypoxia reoxygenation injury.Intracellular calcium concentration,calmodulin(CaM) and calcium/calmodulin dependent kinaseⅡ?(CaMPKⅡ?) mRNA were measured by fluorospectrophotometry and reverse transcription-polymerase chain reaction(RT-PCR)respectively.Results After hypoxia/reoxygenation,intracellular calcium concentration,CaM and CaMPKⅡ? expression were enhanced(P

Aim To explore the effects of shuang shen tong guan(SSTG) drug serum on calcium and NOS-NO system in cardiomyocytes injured by hypoxia and reoxgyenization.Methods The cardiomyocytes were deprived of oxygen and glucose to mimic hypoxia reoxygenation injury.Intracellular calcium concentration was determined by Fluorospectrophotometry.NOS activity and NO contents were detected using colorimetric method.Results Intracellular calcium concentration,NOS activity and NO content were increased after hypoxia and reoxygenation injury.SSTG drug serum inhibted the upwards of intracellular calcium concentration,activity of total NOS,indusive NOS and NO content due to hypoxia/reoxygenation injury significantly(P

Cellular signal transduction plays an important role in pathogenesis of myocardial ischemia reperfusion(MIR) injury. During this process, extracellular signal molecules, membrance receptors and intracellular signal paths are involved. There are many cross talkings in signal pathways at many levels. The present article reviews cellular signal transduction mechanisms in MIR. The exploration of MIR's cellular pathogenesis may be beneficial for us to prevent and cure the heart damage in clinic. Furthermore, it may provide a new way of thinking in mechanism research of medicine.

Tong-Mai(TM) granules were composed ofRadix Salviae Miltiorrhizae(Danshen),Rhizoma Chuanxiong(Chuanxiong) andRadix Puerariae(Gegen). It had the effect of activating blood circulation. It had been used to treat ischemic cardio-cerebrovascular diseases in the clinical practice. This research combined serum pharmacochemistry and serum pharmaocology to study the material basis of active components in TM granules. After single or multiple intragastric administrations of TM granules, serum blood samples of rats were collected at different time points. LC-MS/MS method was developed to analyze chemical components of TM in blood serum samples. The cardiomyocyte hypoxia / reoxygenation (H/R) model was used in the evaluation of cardiomyocyte protection by TM. The correlation analysis was also conducted between serum concentration of TM and cardiomyocyte activity. The results showed that 8 components of pueraria flavonoid, 5 components of salvianolic acids and 2 components fromGegen were promptly absorbed and reached their highest concentrations at 5 or 30 min after administration. After 3 times of medication, the serum concentration was obviously higher compared to single medication. The drug-serum of TM showed significant protective effect on the cardiomyocyte H/R injury with dose-effect relationship. Daidzein, lithospermic acid, salvianolic acid A, salvianic acid A and rosmarinic acid presented as the most correlated components linked to the effect of activating blood circulation by TM. The serum pharmacochemistry / serum pharmacology related studies provided references for the verification of material basis of active components in compound Chinese medicine.

OBJECTIVETo investigate the effects of Shuangshen Tongguan Recipe (SSTG) on myocardial nuclear factor-kappa B (NF-kappaB) signal pathway, expression of myocardial junction intercellular communication (MJIC) connexin 43 (Cx43), and infarcted myocardial size and weight of the rats' heart after acute myocardial ischemia/reperfusion (I/R) damage.

METHODSModel rat of I/R injury was established by coronary arterial ligating/ releasing. The infarcted myocardial size and weight were determined by N-BT staining, expression of NF-kappaB p65 in myocardial tissue and Cx43 were determined by immunohistochemical method, contents of serum tumor necrosis factor-alpha(TNF-alpha) and intercellular adhesion molecule-1 (ICAM-1) were measured by ABC-ELISA.

RESULTSThe myocardial infarcted size and weight, expression of NF-kappaB p65, contents of serum TNF-alpha and ICAM-1 of I/R injured rats in the model group were significantly increased (P<0.05), while Cx43 degraded markedly after modeling. These changes were restored after treated with SSTG (P <0.05).

CONCLUSIONSerious myocardial infarction occurs after ischemia/reperfusion injury, combined with NF-kappaB signal pathway activation and severe Cx43 degradation. SSTG could inhibit the activation of NF-kappaB, the over-excretion of TNF-alpha and ICAM-1 in serum, and the degradation of Cx43 to decrease the myocardial infarcted size and weight.

Animals ; Connexin 43 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Intercellular Adhesion Molecule-1 ; blood ; Male ; Myocardial Reperfusion Injury ; blood ; drug therapy ; Myocardium ; metabolism ; pathology ; NF-kappa B ; biosynthesis ; physiology ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

OBJECTIVETo study the time-effect, dose-effect of serum containing Shuangshentongguan (SSTG), a formula composed by active fraction of Chinese medicine.

METHODSerum containing SSTG was obtained at different time after perorally administrated with different dose of SSTG. The cardiomyocytes were deprived of oxygen and glucose to mimic hypoxia reoxygenation injury and were treated by serum containing SSTG when reoxygenation. LDH content in supernatant was detected after reoxygenation. LDH release suppression rate was used to study the time-effect and dose-effect of serum containing SSTG.

RESULTThe suppression rate of LDH release was not satisfied in serum containing high dose of SSTG. There was no significant dose-effect relationship between them. The suppression rate of LDH release was also different in drug serum taken at different time after SSTG treatment.

CONCLUSIONIt was not the case that the higher the dose given to animal, the better the pharmacological effects.

Animals ; Cell Hypoxia ; Cells, Cultured ; Corydalis ; chemistry ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacokinetics ; Female ; L-Lactate Dehydrogenase ; metabolism ; Male ; Myocytes, Cardiac ; cytology ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Serum ; Time

OBJECTIVETo explore the effects and mechanism of Shuangshen Tongguan (SSTG) serum, a formula composed of the active fractions of Chinese medicine, on calcium overload in cultured cardiomyocytes injured by hypoxia and reoxygenation.

METHODThe cardiomyocytes were deprived of oxygen and glucose to producl hypoxia reoxygenation injuried models. The changes of intracellular calcium fluorescence intensity induced by K+ and Thapsigargin were measured by fluorospectrophotometry and laster scanning confocal microscope respectively.

RESULTIntracellular calcium concentration was low in normal cardiomyocytes and was enhanced after hypoxia/reoxygenation (P < 0.05); SSTC drug serum reduced the intracellular calcium concentration and depressed the increase of calcium fluorescence intensity in singular cardiomyocyte due to K+ and Thapsigargin stimulation.

CONCLUSIONHypoxia/reoxygenation injury, K+ and Thapsigargin could induce calcium overload in cardiomyocytes. The effects of calcium antagonism of SSTG drug serum were achieved by inhibiting calcium inflow, promoting calcium re-absorption of calcium.

Animals ; Animals, Newborn ; Calcium ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Corydalis ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Serum ; Thapsigargin ; antagonists & inhibitors

There is a great need for new vaccine development against influenza A viruses due to the drawbacks of traditional vaccines that are mainly prepared using embryonated eggs. The main component of the current split influenza A virus vaccine is viral hemagglutinin (HA) which induces a strong antibody-mediated immune response. To develop a modern vaccine against influenza A viruses, the current research has been focused on the universal vaccines targeting viral M2, NP and HA proteins. Crystallographic studies have shown that HA forms a trimer embedded on the viral envelope surface, and each monomer consists of a globular head (HA1) and a "rod-like" stalk region (HA2), the latter being more conserved among different HA subtypes and being the primary target for universal vaccines. In this study, we rationally designed the HA head based on the crystal structure of the 2009-pandemic influenza A (H1N1) virus HA as a model, tested its immunogenicity in mice, solved its crystal structure and further examined its immunological characteristics. The results show that the HA globular head can be easily prepared by in vitro refolding in an E. coli expression system, which maintains its intact structure and allows for the stimulation of a strong immune response. Together with recent reports on some similar HA globular head preparations we conclude that structure-based rational design of the HA globular head can be used for subtype-specific vaccines against influenza viruses.
Animals ; Antibodies, Viral ; immunology ; Crystallography, X-Ray ; Drug Design ; Female ; Freund's Adjuvant ; administration & dosage ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; immunology ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; immunology ; Influenza Vaccines ; administration & dosage ; biosynthesis ; Influenza, Human ; immunology ; prevention & control ; virology ; Mice ; Mice, Inbred BALB C ; Models, Molecular ; Protein Folding ; Recombinant Proteins ; genetics ; immunology ; Structure-Activity Relationship ; Vaccination ; Vaccines, Subunit ; administration & dosage ; biosynthesis

Objective:To study the correlation between liver imaging reporting and data system (LI-RADS) category with tumor differentiation, Ki67 index, microvascular infiltration, and predictive prognosis in hepatocellular carcinoma (HCC).Methods:We retrospectively analyzed the clinical and radiological data of 178 patients with HCC who were confirmed by histopathological studies after liver resection between January 2015 and September 2020 at Lishui Central Hospital and Lishui People’s Hospital. There were 156 males and 22 females, with age of (57±10) years old. These patients were assessed for LI-RADS categories according to the 2018 version of LI-RADS, and they were divided into 4 groups according to the assessment results: 12 patients with LI-RADS-3 (the LI-RADS-3 group); 26 patients with LI-RADS-4 (the LI-RADS-4 group); 102 patients with LI-RADS-5 (the LI-RADS-5 group); and 38 patients with LI-RADS-M (the LI-RADS-M group). The patients' general information, tumor markers, pathology and other clinical data were recorded. Correlation analysis between the LI-RADS category with pathology was performed by the Kendall's tau-b test. Survival analysis between groups was performed by the Kaplan-Meier analysis. The Cox regression risk model was used to analyze the relationship between these variables with the risk of death.Results:The Kendall's tau-b test showed that LI-RADS category was positively correlated with the degree of tumor differentiation ( t=0.204, P=0.002), but not with microvascular infiltration and Ki 67 index ( P>0.05). All patients were followed up for 4.2 to 84.2 months (median follow-up 36.3 months). By the end of follow-up, 31 patients had died and 147 patients were alive. The cumulative 1-year and 3-year survival rates of the LI-RADS-5 group were 97% and 90% respectively, which were significantly higher than those in the LI-RADS-M group (81% and 63%), and the LI-RADS-4 group (96% and 81%), ( P<0.05). The cumulative 1-year and 3-year survival rates of patients in the LI-RADS-3 group were 100% and 67% respectively, and there was no statistically significant difference with the LI-RADS-5 group ( P>0.05). The Cox multivariate regression analysis showed that tumor glycoantigen 199 (>50 μl/ml) to be an independent influencing factor in survival of HCC patients ( HR=0.43, 95% CI: 0.24-0.76, P=0.004). Conclusion:The LI-RADS category of HCC was positively correlated with the degree of tumor differentiation, and patients with HCC meeting the LI-RADS-5 criteria had relatively better prognosis.

OBJECTIVETo investigate the cardio-protective effects of Corocalm on acute myocardial ischemia in rats, and to explore its possible therapeutic mechanisms.

METHODSThe acute ischemic model was prepared by ligating the left anterior descending (LAD) coronary artery in rats. The animals were divided into 6 groups, 8 in each group. The sham operated group underwent heart exposure without ligation and were treated with normal saline 3 ml/kg, while the other 5 groups, the model groups, consisted of acceptable acute ischemic model rats and were also treated with normal saline, with the Guanxin Capsule (GXC) group treated with refined GXC, 600 mg/kg, the low and high dose Corocalm groups treated with 85 mg/kg and 340 mg/kg of Corocalm respectively, and the Diltiazem group, treated with Diltiazem 5 mg/kg, with all the tested drugs prepared with normal saline into equal volume (3 ml/kg) and administrated once via duodenum 10 min before ligation. Myocardial infarction area was determined by the quantitative histological assay with nitroblue tetrazolium (N-BT) stain. And the levels of creatine phosphokinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA) content, and the activity of superoxide dismutase (SOD) in serum were measured by biochemical assay and spectrophotometry respectively. Besides, the blood viscosity in another 50 rats was determined, who received for 7 successive days oral administration with different concentration of Corocalm or aspirin.

RESULTSIt showed that low and high dose Corocalm could significantly reduce the infarction area, inhibit the increase of serum CK, LDH activity and MDA content, and enhance the SOD activity after ischemia/reperfusion. The whole blood viscosity at different shear rates in rats treated with high dose Corocalm was significantly lower than those treated with normal saline (P < 0.05).

CONCLUSIONCorocalm has favourable protective effects on heart in ischemic condition, the effect of which might be through its actions in inhibiting CK and LDH activity, scavenging oxygen free radicals, and lowering blood viscosity.

Acute Disease ; Animals ; Blood Viscosity ; drug effects ; Cardiotonic Agents ; pharmacology ; Creatine Kinase ; blood ; Drugs, Chinese Herbal ; pharmacology ; L-Lactate Dehydrogenase ; blood ; Male ; Malondialdehyde ; blood ; Myocardial Infarction ; drug therapy ; metabolism ; pathology ; Myocardial Reperfusion Injury ; drug therapy ; metabolism ; pathology ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; blood