Objective: To develop an HPLC-DAD method for the simultaneous determination of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B, and hydroxysafflor yellow A (HYSA) in cerebral ischemic injury of rats, in order to provide an accurate and reliable analytical method to study the pharmacokinetics. Methods: The middle cerebral artery occlusion (MCAO) model was established, in which all the rats were iv injected with the active constituents in Danhong Injection simultaneously (danshensu, protocatechuic aldehyde, salvianolic acid B, and HYSA). The analysis of the measured components' plasma concentration was carried at 30℃ on the reversed-phase column of Agilent Eclipse XDB-C18 (150 mm×4.6 mm, 5 μm) and eluted with acetonitrile and water containing 0.4% phosphate acid as mobile phase in gradient mode. Detection wavelengths were 280 nm for danshensu, protocatechuic acid, vanillic acid, salvianolic acid B and 403 nm for HYSA. The propyl p-hydroxybenzoate was used as the internal standard (IS). Results: Protocatechuic aldehyde rapidly metabolized in MCAO rats, but its metabolites, protocatechuic acid and vanillic acid, were easily detected. The linear ranges of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B, and HYSA were 0.35-140 (R2=0.9998), 0.15-60 (R2=0.9990), 0.05-20 (R2=0.9988), 0.25-100 (R2=0.9996), and 0.075-30 mg/L (R2=0.9985), respectively. The mean recoveries were between 80% and 120% and the RSD value of intra-day and inter-day was less than 10%. The results of stability meet the requirements for biopharmaceutical analysis. The main pharmacokinetic parameters of danshensu, protocatechuic acid, vanillic acid, salvianolic acid B and HYSA were as follows: AUC0~∞ were (1143.862±230.840), (427.024±59.293), (135.785±47.631), (418.631±66.242), (288.788±87.809) mg·min/L and t1/2z were (71.496±29.067), (82.379±26.279), (40.331±6.006), (125.164±59.709), (177.577±112.836) min. Conclusion: The method is simple, rapid, precise and stable, which can be used to the pharmacokinetic studies on the compatibilities of danshensu, protocatechuic aldehyde, salvianolic acid B, and HYSA in MCAO rats.

OBJECTIVETo investigate the protective effect of extracts from Cichorium endivia (CEE) in H2O2-induced HepG2 cell oxidative stress injury, and explore the antioxidant mechanism of CEE in HepG2 cells.

METHODThe viability of H2O2-induced HepG2 cells and the intracellular ROS level were measured by MTT assay and DCFH-DA fluorescence staining assay. The antioxidant-response element (ARE)-Luciferase activity was tested in HepG2 cells stably transected by ARE reporter gene. The fluorescence quantitative RT-PCR was adopted to determine the mRNA expressions of genes containing ARE sequence in HepG2 cells.

RESULTThe cell viability reduced, while the ROS level increased after HepG2 cells were treated by H2O2. Different concentrations of CEE could be added to significantly improve the above results. After HepG2 cells transected by ARE reporter gene were treated with different concentrations of CEE, the intracellular ARE activity could increase in a concentration-dependent manner. In addition, the mRNA expressions of regulatory genesGCLC, GCLM and HMOX-1 containing ARE sequence in HepG2 cells were up-regulated in a concentration-dependent manner by CEE.

CONCLUSIONCEE inhibited the H2O2-injured HepG2 cells by reducing the ROS level. CEE's antioxidant mechanism for HepG2 cells may be closely related to the antioxidant defense system associated with its effect of activating Nrf2-ARE pathway in HepG2 cells.

Antioxidants ; isolation & purification ; pharmacology ; Asteraceae ; chemistry ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hep G2 Cells ; Humans ; Hydrogen Peroxide ; pharmacology ; Reactive Oxygen Species ; metabolism ; Response Elements ; genetics

ObjectiveTo analyze characteristic changes of shoulder muscles by investigating surface electromyographic regularity changes before and after the treatment of Frozen shoulders. Methods7 cases of frozen shoulders were selected, surface electromyography (sEMG) before and after treatment were recorded by AMT-8 EMG recorder. The collected data was processed using MATLAB software integrated EMG (IEMG) value were obtained. Then the IEMG data were statistically analyzed using Stata11.0 software and compared. Results ①The differences of abduction deltoid and infraspinatus muscle IEMG values before and after treatment were statistically significant(P ＜0.05), while IEMG values changes of biceps, triceps, pectoralis major, latissimus dorsi and trapezius before and after treatment were not statistically significant (P＞0.05); changes of adduction IEMG values of all muscles were not statistically significant (P＞0.05). ②Changes of flexion IEMG values of biceps before and after treatment were statistically significant (P＜0.05), while IEMG values changes of pectoralis major, infraspinatus muscle, latissimus dorsi, trapezius, deltoid, triceps before and after treatment were not statistically significant (P＞0.05). Changes of extension IEMG values of triceps before and after treatment were statistically significant (P＜0.05), while those of pectoralis major, infraspinatus muscle, latissimus dorsi, trapezius, deltoid and biceps were not statistically significant (P＞0.05). ③IEMG values of all the muscles during external rotation, internal rotation before and after treatment IEMG were not significantly different. Conclusion①After treatment, the outreach functions of infraspinatus and deltoid muscles were improved, while adduction functions of all muscles were not improved. ②Flexion function of biceps was improved significantly after treatment as well as extension function of triceps.③Extemal rotation, internal rotation functions of all muscles were not improved significantly.

To study the pharmacokinetic process of Danshensu in cerebal ischemia injury model rats and the correlation with its anti-cerebral ischemia effect. In this study, the middle cerebral artery occlusion (MCAO) model was established, in which all of the rats were intravenously injected of Danshensu at a single dose of 40 mg x kg(-1). The HPLC-DAD method was applied to determine the plasma concentration of Danshensu at different time points and draw the drug-time curve. Meanwhile, the superoxide dismutase (SOD) and the lactate dehydrogenase (LDH) activity were determined to draw the time-effect curve. The DAS 3.2. 6 software was used to process the data, analyze their correlation, compare the pharmacokinetic difference between model and normal rats after the administration of the same doses of Danshensu and the changes in pharmacodynamic indicators of model rats after the administration, and evaluate the effect of Danshensu in treating the cerebral ischemia disease. According to the results, the pharmacokinetic processes of Danshensu in the cerebral ischemia-reperfusion and normal rats were consistent to the two-compartment model. The main pharmacokinetic parameters were: t1/2alpha were (0.267 +/- 0.026), (0.148 +/- 0.020) h;t1/2beta were (1.226 +/- 0.032), (1.182 +/- 0.082) h; AUC0-infinity were (42.168 +/- 4.007), (26.881 +/- 1.625) mg x L(-1) x h. After the cerebral ischemia-reperfusion, the activity of SOD decreased and the activity of LDH increased. Danshensu could inhibit the decrease in the SOD activity and the increase in the LDH activity within a certain period of time. This indicated that Danshensu could stay longer in cerebral ischemia-reperfusion rats than in normal rats and eliminated more slowly, which reflected the rationality of Danshensu in the clinical treatment of cerebral ischemia diseases. Danshensu's effect against the cerebral ischemic injury may be related with its level in vivo. Its plasma concentration is positively related to the SOD activity and negatively related to the LDH activity.
Animals ; Brain Ischemia ; drug therapy ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry

AIMTo explore the effects of aminoguanidine (AG) inhalation on bleomycin (BLM)-induced fibrosis in lungs of rats and its possible mechanism.

METHODSSixty male Sprague-Dawley rats were randomly divided into 4 groups: BLM plus normal saline (NS) group, BLM plus 10 mmol/L AG group, BLM plus 50 mmol/L AG group, and NS plus NS group. At the same day when administrated by single intratracheal instillation of BLM (5 mg/kg) or equal volume of NS as control, the rats received NS (the same volume as AG) or AG inhalation (10 mmol/L AG, or 50 mmol/L AG, 5 min/each time, 2/day) for 30 d. The nitrite/nitrate (NO2-/NO3-) content of plasma in pulmonary artery, hydroxyproline content and the pathological changes in lungs, as well as lipid peroxide (LPO) content of plasma in pulmonary artery were examined.

RESULTSThe NO2-/NO3- content of plasma in pulmonary artery was increased in rats on day 14 after intratracheal instillation of BLM, compared with that of the control rats (P < 0.01). The hydroxyproline content in lung, the grade of pulmonary alveolitis and the content of LPO of plasma in pulmonary artery were increased in rats on day 30 after intratracheal instillation of BLM, compared with that of the control rats, respectively (P < 0.05, P < 0.01, P < 0.01). The above-mentioned changes were ameliorated by AG inhalation (10 mmol/L AG, or 50 mmol/ LAG, 5 min/each time, 2/day) for 30 d (P < 0.01, P < 0.05, P < 0.05, P < 0.01).

CONCLUSIONAG inhalation has anti-action on BLM-induced fibrosis in lung, which might be related to blockage of oxidative injury in lung.

Administration, Inhalation ; Animals ; Bleomycin ; Enzyme Inhibitors ; administration & dosage ; pharmacology ; Guanidines ; administration & dosage ; pharmacology ; Male ; Nitric Oxide Synthase ; antagonists & inhibitors ; Oxidative Stress ; drug effects ; Pulmonary Fibrosis ; chemically induced ; physiopathology ; prevention & control ; Rats ; Rats, Sprague-Dawley

Objective:To explore the type distribution and severity of central nervous system(CNS) complications in severe hand, foot and mouth disease (HFMD) cases prior to the introduction of human enterovirus A group 71 type (EV-A71) vaccine, and provide scientific data for early clinical intervention.Methods:A total of 3 583 laboratory-confirmed severe HFMD cases in Jiangsu province during 2010-2016 were collected and analyzed retrospectively. Related early warning signs of increased HFMD severity were estimated with logistic regression analyses.Results:The severity-fatality rate, severity-pediatric intensive care unit(PICU) admission rate, and sequelae rate were 8.09‰(29/3 583), 11.75% (421/3 583)and 5.30‰(19/3 583). Of them, 39.02% (1 398/3 583) patients suffered from mild CNS involvement, 59.22% (2 122/3 583) patients suffered from severe CNS involvement, 1.76%(63/3 583) suffered from critical CNS involvement. The rates of the cases whose age of onset was 6-11 months, the rates of cases with atypical rash, respiratory-related signs and symptoms (shortness of breath, slowed breathing, dyspnea, etc), neurological-related signs and symptoms [(hand and foot shaking, convulsions, lethargy(sleepiness), etc], circulatory-related system signs and symptoms (faster heart rate, abnormal skin color, arrhythmia, cold limbs), laboratory-related indicators (increased white blood cell count, increased lymphocyte count, increased platelet count, increased C-reactive protein, etc), clinical auxiliary examination [electroencephalogram(EEG), brain CT, chest X-ray)] were highest in the critical CNS involvement group, and the differences were significant ( P<0.05). Multivariate logistic regression analysis showed that with the increase of proportion of convulsions, slowed breathing, vomiting, meningeal irritation and other 7 variables, the severity of CNS complications increased ( P<0.05). Conclusions:The indicators such as easily startled, slow breathing, vomiting, elevated lymphocytes, abnormal EEG and other indicators have important clinical significance for children with severe HFMD to progress to CNS complications of different severity.

OBJECTIVETo study the effects of different parameters (frequency, intensity, needle-retained time and treatment interval) of electroacupuncture at Fenglong (ST 40) for adjusting blood lipids, so as to find out the optimization parameter.

METHODSFifty-four cases meeting the criteria for hyperlipoidemia were randomly divided into 27 groups with orthogonal design L27 (3(13) ). According to the orthogonal design program they were treated with electroacupuncture at Fenglong (ST 40). Ten sessions constituted one course with a one week's interval between two course. The treatment was given for 2 courses.

RESULTS(1) The parameters of EA at Fenglong (ST 40) for regulating blood lipids in primary and secondary orders are: frequency, needle-retained time, interval of treatment, intensity. (2) The parameters of EA at Fenglong (ST 40) for various programs in regulating various blood lipids are: for TG, frequency AM 50 Hz, needle-retained time 20 ain, intensity 1 mA, twice each week; for TC, frequency AM 100 Hz, needle-retained time 30 min, intensity 1 mA, once every other day; for LDL-C, frequency Am 100 Hz, needle-retained time 30 min, intensity tolerable and comfortable, once every other day.

Acupuncture Points ; Aged ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Electroacupuncture ; methods ; Female ; Humans ; Hyperlipidemias ; blood ; therapy ; Lipids ; blood ; Male ; Middle Aged

Aim To determine the effect of exosomes from lipopolysaccharide-treated human bone marrow mesenchymal stem cells on proportion of Ly6Chigh and Ly6Clow monocytes/macrophages in inflammatory micro- environment. Methods BMSCs were obtained by gra-dient centrifugation, identified and then treated with li-popolysaccharide for 48 h. The exosomes were purified from conditional medium with or without LPS treatment and identified by CD63 protein using Western blot and transmission electron microscope. The diameters and concentration were detected by Nanoparticle Trafficking Analysis ( NTA ) . The monocytes/macrophages were sorted from bone marrow of the mice by magnetic beads. Cells were co-cultured with exosomes for 24 hours, and then treated with LPS for 48 hours. The proportion of Ly6C monocytes/macrophages was detec-ted by flow cytometry. Inflammatory cytokines in cell supernatant were investigated using ELISA. Results BMSCs surface markers CD44, CD90 were positively detected, but CD34, CD45 were not expressed. BM-SCs presented adipogenic differentiation ability. Exo-somes were positively expressing CD63 protein, and NTA showed that the diameters of exosomes were up to (82.4 ± 3.7 ) nm. BMSCs stimulated by LPS pro- duced more exosomes ( P < 0.01 ) . Exosomes from BMSCs with or without LPS treatment could increase the ratio of Ly6Chigh monocytes (P<0.01) and down-regulate the ratio of Ly6Chigh macrophages (P<0.05), and the effect of LPS treated-exosomes was more signif-icant than untreated-exosomes (P<0.05). Moreover, the concentration of IL-6 was also elevated under exo-somes treatment ( P <0.05 ) . Conclusions Human bone marrow mesenchymal stem cells-derived exosomes contribute to the regulation of Ly6Chigh monocytes/mac-rophages, indicating that they could be involved in the therapeutic treatment of inflammatory diseases.

Medicinal botanical gardens play important roles in promoting the development of TCM industry. With the advancing of the construction of the national medicinal plant garden system, the upgrading construction work for Huazhong Medicinal Botanical Garden, one of its members, is grasping to be carried on. While it is believed that there is a tremendous potential development in the combination of selenium and Chinese herbal medicine. In this article, the significance of selenium for Huazhong Medicinal Botanical Garden was investigated and then the routes to highlight selenium characteristics in its upgrading construction were put forward as follows for some responsibility department as reference: (1) Concentrating on the background investigation and conserving selenium resources;(2) Selecting and breeding special germplasm materials associated with selenium for selected key species; (3) Aiming at the specific efficiency and screening selenium-enriched medical plants; (4) Selecting large varieties of TCM and carrying out the research of selenium-rich technical ways of artificial cultivation; (5) Propelling the research and development of selenium-containing health products, based on resources industrialization; (6) Constructing new specialized garden for selenium-enriched medical plants.

OBJECTIVETo evaluate the disadvantage of nonstandard operation for thyroid papillary cancer, and the value of re-operation.

METHODSEight hundred and ninety three thyroid papillary cancer patients (332 patients who received nonstandard operation received re-operation and 561 patients who received first standard operation in Zhejiang Cancer Hospital from January 1990 to January 2000) were retrospectively reviewed.

RESULTSPathological results confirmed that there were 53.9% cases with residual cancer in re-operative specimen. The parathyroid was preserved in 74.1% cases in re-operation and 93.0% cases in standard operation (P < 0.01). The recurrent laryngeal nerve was injured in 3.3% cases in reoperation and 1.2% cases in standard operation (P < 0.05). The positive neck lymph node was 39.2% and 37.4% (P > 0.05), the 5-year local recurrence rate was 7.5% and 3.7% (P < 0.01), the total 5-year and 10-year cum-survival rates were 90.2%, 84.4% in patients with re-operation and 94.0%, 92.5% in patients with standard operation. The 10-year cum-survival rate were 67. 8% in patients with recurrence and 92. 9% in patients with non-recurrence in primary site respectively. Statistical analysis showed that the cancer recurrence influences the survival rate significantly (P < 0.01).

CONCLUSIONSIn consideration of the higher residual tumor rate in patients who received nonstandard operations, the re-operations were necessary. But the re-operation could also leads to higher recurrence rate and more functional injuries. The standardized primary operation should therefore be formulated and advocated.

Adolescent ; Adult ; Aged ; Carcinoma, Papillary ; surgery ; Child ; Female ; Humans ; Male ; Middle Aged ; Reoperation ; Thyroid Neoplasms ; surgery ; Thyroidectomy ; methods ; Young Adult