Objective To observe the protective effect of different doses of left carnitine on cardiomyocyte function in children with viral myocarditis.Methods94 cases of children with viral myocarditis were selected and divided into observation group and control group according to the random number table, 47 cases in each group.Both groups were treated with fructose 1,6-diphosphate(100~250mg/kg, add 150mL of 10% glucose in intravenous infusion, 1times/d) combined with left carnitine.Large dose group of left carnitine 100 mg/kg, low dose group 50mg/kg, all added 150mL of 5% glucose intravenous infusion, 1 times/d.The total effective rate, creatine kinase(CK), creatine kinase isoenzyme (CK-MB),lactate dehydrogenase (LDH), cardiac ejection fraction(EF), ventricular short axis shortening (FS) And the total incidence of adverse reactions were compared between two groups.ResultsThe total effective rates of two group were 93.62% and 87.23%.The level of CK,CK-MB,LDH in two groups were significantly decreased after treatment of two weeks(P<0.05), EF, FS were increased after treatment of two weeks(P<0.05).There was no significant difference in CK, CK-MB, LDH, EF and FS between the high dose group and the low dose group after treatment of two weeks.The overall incidence of adverse reactions in the high dose group was 25.53%, which was lower than that in the low dose group (8.51%,P<0.05).ConclusionThe use of low-dose left carnitine in children with viral myocarditis can effectively remove free radicals, protect cardiomyocyte function and improve myocardial energy metabolism and cardiac function, and safer than high-dose groups.

Objective To explore the clinical features and treatment of children with acute severe viral myocarditis.Methods The clinical data of presentation,diagnosis,therapy and prognosis of children who were admitted in our hospital from Jan 2005 to Jan 2012 with acute severe viral myocarditis(severe myocarditis group) were analyzed retrospectively.Twenty-three cases of normal healthy children in the same period were selected as control group.The levels of serum cardiac troponin(CTn)-Ⅰ and N-terminal pro-brain natriuretic peptide(NT-proBNP) were detected by ELISA method,the changes of left ventricular ejection fraction and left ventricular fraction shortening were understood by color doppler echocardiography.Results The level of CTn-Ⅰin severe myocarditis group was significantly higher than that of control group,the difference was statistically significant [(18.67 ± 12.31) ng/ml vs (0.02 ±0.01) ng/ml,P ＜0.05].Compared with the acute phase,the level of CTn-Ⅰshowed a trend of gradual decline in 7 d [(0.55 ±0.24) ng/ml],basic close to normal in 14 d [(0.06 ±0.03) ng/ml] (P ＜0.05).The level of NT-proBNP increased significantly in severe myocarditis group compared with control group [(3 067.26 ± 902.79) pg/ml vs (80.04 ± 17.79) pg/ml,P ＜0.05].Compared with acute phase,the levels of NT-proBNP were closed to normal in 7 d [(648.63 ±342.37) pg/ml] and 14 d [(213.58 ± 129.51) pg/ml] (P ＜ 0.05).The left ventricular ejection fraction [(52.63 ± 6.98) ％ vs (71.39 ± 2.41) ％] and left ventricular fraction shortening [(32.1 ± 2.97) ％ vs (40.04 ± 2.31) ％] in severe myocarditis group were significantly lower than those in control group (P ＜ 0.05).Conclusion Acute severe viral myocarditis of children was characterized by rapid onset,severe illness and high mortality.Early use of adrenal cortical hormone and gamma globulin under the comprehensive treatment and application temporary pacemaker can help patients to recover from the disease.

Objective: To explore the impact of hydrogen sulfide (H₂S) on the heme oxygenase-1/carbon monoxide pathway in Coxsackie virus B3 (CVB3)-induced murine myocarditis (VMC) model. Method: A total of 70 inbred male Balb/c mouse (4-6 weeks old) were randomized into the following four groups: Normal, VMC, PAG and NaHS (n=10 for Normal, n=20 for VMC, PAG and NaHS groups). Mice in Normal group were non-infected mice treated with intraperitoneal injection of sterile phosphate-buffered saline daily for 10 days.Mice in VMC group received intraperitoneal CVB3 injection (0.1 ml 10^-5.69TCID₅₀m·ml^-1·d^-1 and PBS for 10 days), and mice in PAG group received additional intraperitoneal DL-proparglygylcine injection (40 mg·kg^-1·d^-1 for 10 days), mice in NaHS group received additional intraperitoneal NaHS injection (50 μmol·kg^-1·d^-1 for 10 days). All mice were sacrificed on day 10th, and body weight and heart weight, the ratio of heart weight to body weight were compared among groups.Pathological changes of heart tissues were observed microscopically by HE and the histopathologic scores were valued.The content of COHb was tested after the gathering of blood specimens while reverse transcription-polymerase chain reaction was used to detect myocardial HO-1 mRNA expression. Results: (1) Pathological findings in myocardium: hearts sections in Normal group were normal and no inflammatory cells and necrosis were found.A notable cellular infiltration, interstitial edema, vascular hyperemia and necrosis were observed in heart section of VMC, PAG and NaHS group.Extensive inflammations and larger area of myocardial cells necrosis were evidenced in PAG group and above changes were significantly reduced in NaHS group.(2) Comparison of the ratio of heart weight to body weight and histological scores of myocardium: the ratio was significantly higher in the VMC, PAG, NaHS groups than in Normal group (P<0.05), which was higher in PAG group and lower in NaHS group as compared with VMC group (both P<0.05). The histopathologic scores of all CVB3 innoculation groups were higher than in the Normal group, which was higher in PAG group and lower in NaHS group as compared to VMC group (both P<0.05). (3) The content of blood COHb in VMC, PAG or NaHS group was significantly higher than that in Normal group (P<0.05), which was significantly lower in PAG group, and higher in NaHS group as compared to VMC group (both P<0.05). (4) The mRNA expression of myocardial HO-1 detected by RT-PCR: weak expression was observed in Normal group, which was significantly upregulated in VMC, PAG and NaHS groups (P<0.05), which was downregulated in PAG group and upregulated in NaHS group as compared to VMC group (both P<0.05). (5) Correlation analysis: blood COHb concentration was positively correlated with myocardial HO-1 mRNA expression(r=0.927, P=0.000), negatively correlated with histopathologic scores(r=-0.753, P=0.000)and the histopathologic scores were negatively correlated with the myocardial HO-1 mRNA expression (r=-0.754, P=0.000). Conclusions H₂S could play a protective role in murine CVB3 myocarditis model through inducing HO-1 expression and upregulating HO-1/CO pathway.

OBJECTIVETo explore the effects of tanshinone and JAK2/STAT1 signaling pathway related mechanism in CVB3-induced myocarditis in murine.

METHODSA total of 110 inbred male Balb/c mice which were 4 to 6 weeks-old were randomly divided into five groups: normal control (N, n = 10), myocarditis control (C, n = 25), tanshinone group (T, 15 mg · kg⁻¹ · d⁻¹, i.p., n = 25), janus kinase 2 inhibitor AG490 group (A, 10 mg · kg⁻¹ · d⁻¹, i.p., n = 25), T+A group (H, n = 25). Myocarditis was induced by 0.5 ml 10(-9.51) TCID50/ml CVB3 i.p. injection for 10 days in group C, T and H. Myocardial histopathologic changes were observed and phospho-STAT1 expressions were detected by immunohistochemistry and Western blot analysis. The levels of serum cardiac troponin I were detected with chemiluminescence immunoassay.

RESULTS(1) Compared with group C, the histopathologic scores were significantly higher in group A and H (3.35 ± 0.57 and 3.34 ± 0.54 vs. 2.12 ± 0.39, P < 0.01), but lower in group T (1.40 ± 0.34 vs.2.12 ± 0.39, P < 0.01). (2) The expression of p-STAT1 protein was similar in group A and H compared to group N (P > 0.05), but was significantly lower than that in group C (0.017 ± 0.010 and 0.020 ± 0.010 vs. 0.246 ± 0.010, P < 0.01). The expression of p-STAT1 protein was significantly higher in group T than in group C (P < 0.01). (3) The levels of serum cardiac troponin I in group C, A, T and H were significantly higher than in group N ((0.42 ± 0.06), (1.17 ± 0.25), (0.23 ± 0.05) and (1.04 ± 0.19) µg/L vs. (0.02 ± 0.01) µg/L, all P < 0.01). The levels of serum cardiac troponin I were significantly higher in group A and H compared with group C ((1.17 ± 0.25) and (1.04 ± 0.19) µg/L vs. (0.42 ± 0.06) µg/L, P < 0.01), but were significantly lower in group T than in group C ((0.23 ± 0.05) µg/L vs. (0.42 ± 0.06) µg/L, P < 0.01). (4) There was a negative correlation between the expression level of p-STAT1 and the histopathologic scores (y = -4.503 x + 3.371, R² = 0.738, P < 0.01), but a positive correlation between the levels of serum cardiac troponin I and the histopathologic scores (y = 1.935x + 1.165, R² = 0.766, P < 0.01).

CONCLUSIONTanshinone could attenuate myocardial injury via upregulating the JAK2/STAT1 signaling pathway in this murine viral myocarditis model.

Animals ; Blotting, Western ; Coxsackievirus Infections ; Disease Models, Animal ; Diterpenes, Abietane ; pharmacology ; Heart Injuries ; Janus Kinase 2 ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; drug therapy ; virology ; Myocardium ; STAT1 Transcription Factor ; drug effects ; Signal Transduction ; Troponin I

OBJECTIVE：To study “Qi-invigorating”effect and its possible mechanism of total saponins of Astragalus membranaceus on rats with Qi-deficiency ，and to provide reference for elucidating the material basis of “Qi-invigorating”effect of A. membranaceus . METHODS ：Forty male Wistar rats were randomly divided into normal group ，model group ，positive control group [Buzhong yiqi pills ，4.5 g/（kg·d）]，A. membranaceus total saponins high-dose and low-dose groups [ 252，28 g/（kg·d），by the amount of total saponins] according to body weight ，with 8 rats in each group. Except for normal group ，the model of Qi-deficiency was made in other groups by the method of “diet disorder+fatigue ”. At the same time ，administration groups were given relevant medicine intragastrically ，and normal group and model group were given constant volume of water，once a day ，for consecutive 21 days. After last administration ，the general situation of rats was observed ；the body weight ，spleen index and thymus index of rats were detected ；weight-bearing swimming time was recorded ；the levels of spleen T lymphocyte subsets CD 3 and CD 4，the levels of ATP and ADP in liver tissue ，serum levels of ALB ，RBC and HBG in blood as well as the serum levels of SOD，MDA，lactate，LDH，CK，IL-2，IL-12 and TNF-α were all detected. RESULTS：Compared with normal group ，body weight，thymus index ，spleen index ，weight-bearing swimming time ，the level of spleen T lymphocyte subsets CD 3，ATP，ADP， ALB，IL-2 and IL- 12 were decreased or shortened significantly in model group （P＜0.05 or P＜0.01）. The levels of MDA ， lactate，CK and TNF-α were increased significantly （P＜ 0.05）. Compared with model group ，body weight ，spleen index，weight-bearing swimming time ，the level of spleen T lymphocyte subsets CD 3 and the levels of ATP ，ADP，ALB， RBC and IL- 2 were increased significantly or prolonged（P＜0.05）；while the levels of MDA ，lactate，CK and TNF-α were decreased significantly in A. membranaceus total saponins high-dose group（P＜0.05 or P＜0.01）.Weight-bearing swimming time ，the levels of ATP ，ADP and IL- 2 in A. membranaceus total saponins low-dose group were increased significantly or prolonged （P＜0.05 or P＜0.01），while the levels of MDA ，lactate，CK and TNF-α were decreased significantly （P＜0.05 or P＜0.01）. Compared with positive control group ，spleen index ，spleen T lymphocyte subsets CD 3，weight-bearing swimming time and ATP level of A. membranaceus total saponins high-dose group were increased significantly or prolonged （P＜0.05 or P＜0.01），while MDA levels of A. membranaceus total saponins high-dose and low-dose groups were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：A. membranaceus total saponins can reduce the body ’s accumulation of blood lactic acid ，the activity of CK ，the level of lipid peroxide and regulate immunity to tonify Qi ，delay fatigue and improve exercise ability.

OBJECTIVE：Compare the difference of total protein content of Poria cocos from different producing areas. METHODs：Using bovine serum albumin （BSA） as control ，0.4 mol/L sodium hydroxide solution as exctraction solution ， Coomassie brilliant blue G- 250 as chromogenic reagent ，visible spectrophotometry at 595 nm was used to determine the contents of total protein of P. cocos ；cluster analysis was used to classify 34 batches（S1-S34）of P. cocos from different producing areas. RESULTS：The linear range of BSA was 1.45-17.40 μ g/mL （r＝0.999 6）. RSDs of precision ，stability （20 min） and reproducibility tests were all lower than 3％；recoveries were 100.14％-104.26％（RSD＝1.43％，n＝9）. The contents of total protein in 34 batches of P. cocos from different producing areas were 0.388 4％-1.129 7％. The results of cluster analysis showed that among 34 batches of P. cocos ，the total protein content of P. cocos produced in Yingshan county of Hubei province （S2，S3） was higher than 1％，clustered into one category ；the total protein contents of P. cocos produced in Hubei ，Yunnan，Anhui and Hunan（S1，S5-S10，S12，S13，S16，S17，S19-S21，S23-S25，S28，S30，S31）were 0.653 5％-0.946 1％，clustered into one categony，and the remaining batch content were 0.388 4％-0.601 2％，clustered into one category. CONCLUSIONS ：Established method is suitable for the content determination of total protein content of P. cocos . The protein content of P. cocos from Yingshan county of Hubei province is the highest ，followed by Yunnan and Anhui in 34 batches of P. cocos from different producing areas.

OBJECTIVE：To establis h the UPLC fingerprint of Poria co cos aqueous extract ，and to investigate its relationship with sedative and hypnotic effect. METHODS ：Ten batches of P. cocos from different areas were extracted with water to obtain the aqueous extract. UPLC method was adopted. The determination was performed on Waters HSS-C 18 column with mobile phase consisted of 0.1％ phosphoric acid solution-acetonitrile-methanol （gradient elution ） at the flow rate of 0.4-0.2 mL/min. The detection wavelengths were set at 210 and 242 nm. The column temperature was 40 ℃，and sample size was 2 μL. The fingerprints of 10 batches of P. cocos aqueous extracts were established by using the Similarity Evaluation System of TCM Fingerprint （2012A version），and the common peaks were identified. The sedative and hypnotic effects of 10 batches of P. cocos aqueous extracts from different areas under the synergistic action of pentobarbital sodium were investigated by taking the sleeping rate ，sleep latency and sleep duration of mice as the single efficacy index. After data transformation of single efficacy index and total efficacy （single indexes calculated by analytic hierarchy process ），grey correlation analysis was used to analyze the correlation between the common peaks in fingerprint of P. cocos aqueous extract and the single efficacy index and total efficacy. RESULTS ：There were 24 common peaks in 10 batches of aqueous extract of P. cocos ， and 11 components were identified ， i.e. 16 α-hydroxydehydrotrametenolic acid （peak 6），16α-hydroxytrametendic acid （peak 7），poricoic acid B （peak 9），dehydrotumulosic acid（peak 10），poricoic acid A （peak 12），polyporenic acid C （peak 15），3-O-acetyl-16α-hydroxydehydrotrametenolic acid （peak 17），dehydropachymic acid （peak 20），pachymic acid （peak 21），dehydrotrametenolic acid （peak 22），dehydroeburicoic acid （peak 24）. Grey correlation analysis showed ，the correlation between 24 peaks and sleep duration was greater than 0.6（0.611 5- 0.811 8）；the correlation between 24 peaks and sleep latency was greater than 0.6（0.605 9-0.790 4），except for peaks 14，24 and 2；the correlation of 24 peaks between sleeping rate was greater than 0.6（0.606 4-0.721 6），except for peaks 23，19，17 and 5； the correlation of 24 peaks between total efficacy was greater than 0.6（0.619 0-0.781 2），except for peaks 2，5，19. The top 10 chromatographic peaks related to the total efficacy were peak 15（polyporenic acid C ），peak 16，peak 8，peak 11，peak 12 （poricoic acid A ）， peak 1， peak 7 （16 α-hydroxytrametendicacid）， peak 3， peak 9 （poricoic acid B ） and peak 20 （dehydropachymic acid ）. CONCLUSIONS ：UPLC fingerprint of P. cocos aqueous extract was established and 11 components were identified. Ten components such as polyporus acid C are closely related to the total efficacy of sedation and hypnosis ，which preliminarily reveal the material basis of the sedative and hypnotic effect of P. cocos .

Poria is an important medicine for inducing diuresis to drain dampness from the middle energizer. However, the specific effective components and the potential mechanism of Poria remain largely unknown. To identify the effective components and the mechanism of Poria water extract (PWE) to treat dampness stagnancy due to spleen deficiency syndrome (DSSD), a rat model of DSSD was established through weight-loaded forced swimming, intragastric ice-water stimulation, humid living environment, and alternate-day fasting for 21 days. After 14 days of treatment with PWE, the results indicated that PWE increased fecal moisture percentage, urine output, D-xylose level and weight; amylase, albumin, and total protein levels; and the swimming time of rats with DSSD to different extents. Eleven highly related components were screened out using the spectrum-effect relationship and LC-MS. Mechanistic studies revealed that PWE significantly increased the expression of serum motilin (MTL), gastrin (GAS), ADCY5/6, p-PKAα/β/γ cat, and phosphorylated cAMP-response element binding protein in the stomach, and AQP3 expression in the colon. Moreover, it decreased the levels of serum ADH, the expression of AQP3 and AQP4 in the stomach, AQP1 and AQP3 in the duodenum, and AQP4 in the colon. PWE induced diuresis to drain dampness in rats with DSSD. Eleven main effective components were identified in PWE. They exerted therapeutic effect by regulating the AC-cAMP-AQP signaling pathway in the stomach, MTL and GAS levels in the serum, AQP1 and AQP3 expression in the duodenum, and AQP3 and AQP4 expression in the colon.
Animals ; Rats ; Poria ; Spleen ; Albumins ; Chromatography, Liquid ; Cyclic AMP Response Element-Binding Protein

OBJECTIVE：To study the repa ir，anti-inflammatory and analgesic effects of Compound crocodile oil burn ointment on superficial second-degree burned skin. METHODS ：The heated weight was attached to the right depilated skin of guinea pigs for 4 s to induce the model of superficial second-degree burn. After modeling ，guinea pigs were randomly divided into model group ， Jingwanhong ointment group （positive control ），formula Ⅰ，Ⅱ and Ⅲ groups of Compound crocodile oil burn ointment （volume fraction 1.5％，3％，4.5％，hereinafter），with 8 guinea pigs in each group. Except for model group ，other groups were smeared with 0.7 g/guinea pigs twice a day for 14 consecutive days. The wound healing was recorded every day ，the healing rate of wound was calculated. HE staining was used to observe the histopathological changes of the wound. The serum levels of EGF ，VEGF， SOD，MDA，TNF-α and IL-1 were detected by ELISA. Eighty Kunming mice were divided into 2 groups，and then sub-grouped into model group ，Jingwanhong ointment group （positive control ），formula Ⅰ and Ⅲ groups of Compound crocodile oil burn ointment，with 10 mice in each group. Then xylene auricle swelling method and acetic acid writhing method were used to investigate the anti-inflammatory and analgesic effects of Compound crocodile oil burn ointment. RESULTS ：In the burn repair experiment，after intervention of Compound crocodile oil burn ointment ，the wound area of guinea pigs gradually decreased ，and on the 14th day ，the wound had healed greatly ，and the wound healing rate increased significantly （P＜0.01）；serum levels of EGF and SOD were increased significantly （P＜0.01），while the levels of VEGF ，MDA，TNF-α and IL-1 were decreased significantly（P＜0.05 or P＜0.01）. The thick new epidermal layer was found in wound tissue ，and the connective tissue and neovascularization in the dermis increased significantly. In the anti-inflammatory and analgesic experiment ，after intervention of Compound crocodile oil burn ointment ，the degree of ear swelling and the times of writhing decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS ：Compound crocodile oil burn ointment shows good skin repair ，anti-inflammatory and analgesic efficacy；the mechanism may be associated with increasing the serum levels of EGF and SOD and reducing the levels of VEGF ， MDA，TNF-α，IL-1.