BACKGROUND: Cardiac fibrosis, which results from the loss of balance between synthesis and degradation of cardiac matrix component, is the structural foundation of the stiffness of damaged myocardial tissues. Astragalus membranaceus, a traditional Chinese herb, has multiple functions such as exerting a tonic effect on the heart to induce diuresis. However,the effect of astragaloside Ⅳ on cardiac collagen is poorly known in practice.OBJECTIVE: To observe the effects of astragaloside Ⅳ on myocardial collagen and cardiac function in ischemic rats and to investigate the dosage and time effects of astragaloside Ⅳ.DESIGN: A completely randomized grouping design and randomized controlled trial.SETTING: Department of Pediatrics, the Affiliated Hospital of Medical College of Qingdao University.MATERIALS: The study was conducted in the Encephalopathy Research Institute, Medical College of Qingdao University, from July 2003 to February 2004. Totally 132 Wistar rats of cleaning grade were randomized into three groups: control group (n=11), ischemic group (n=10) and astragaloside Ⅳ group (n=121).METHODS: Rats in control group had thoracotomy, but did not have their left anterior descending coronary artery ligated; rats in ischemic group had thoracotomy and had their left anterior descending coronary artery ligated to establish acute myocardial infarction model; rats in astragaloside Ⅳ group were given astragaloside Ⅳ after surgical ligature of left anterior descending coronary artery. The changes in hemodynamic parameters, cardiac function and myocardial collagen were determined. The dosage and time effects of astragaloside Ⅳ on myocardial collagen and cardiac function were observed.MAIN OUTCOME MEASURES: ① The dosage and time effect of astragaloside Ⅳ on the content of myocardial collagen in the left ventricle of rats with myocardial infarction; ② The dosage and time effects of astragaloside Ⅳ on hemodynamics and cardiac function of rats with myocardial infarction.RESULTS: One hundred rats entered the results analysis. There were 10 in control group and ischemic group, respectively, and 80 in astragaloside Ⅳ group. The five dosage groups of astragaloside Ⅳ [2.5, 5.0, 10.0, 15.0 and 20.0 mg/(kg·d)] and the five postoperative time points (3, 7, 14, 21 and 28 days) had eight rats for each. Astragaloside Ⅳ at a dose of 15.0 mg/kg per day was found to have the most marked effect on ischemic myocardium, so this dose was chosen for observing time effect. ① After administration of astragaloside Ⅳ, the content of collagen in myocardial tissues of the infarcted area of left ventricle, the serum concentration of carboxyterminal procollagen type Ⅰ propeptide and aminoterminal procollagen type Ⅲ propeptide decreased gradually with the increased dose of astragaloside Ⅳ and with the prolonged action time of astragaloside Ⅳ [15 mg/(kg·d)] (P ＜ 0.05-0.01). The serum concentration of carboxyterminal procollagen type 1 propeptide and aminoterminal procollagen type Ⅲ propeptide returned to the level of control at a dose of 10 mg·kg-1per day and at 21 days after astragaloside Ⅳ administration, respectively. The content of collagen in myocardial tissues of the infarcted area of left ventricle was higher than that of non-infarcted area (P＜ 0.01); there were no significant changes in the content of cardiac collagen of right ventricle and non-infarcted area of left ventricle before and after astragaloside Ⅳ administration. ② The cardiac function of ischemic rats significantly improved after astragaloside Ⅳ administration (P ＜ 0.05-0.01); cardiac output, heart rate, stroke volume,mean aortic pressure, systolic aortic pressure, and the stroke work of left ventricle gradually returned to the level of control with the increased dose of astragaloside Ⅳ and with the prolonged action time of astragaloside Ⅳ.CONCLUSION: Astragaloside Ⅳ can inhibit the proliferation of cardiac collagen and improve cardiac function in rats with myocardial infarction.The content of myocardial collagen gradually decreases and cardiac function gradually improves with the increased dose of astragaloside Ⅳ and the prolonged action time of astragaloside Ⅳ.

Objective To explore the effects of recombinant human growth hormone(rh-GH) on immune function in children with tetralogy of Fallot (TOF) after radical operation. Methods Thirty children with TOF were divided into two groups: conventional (n=20) and rh-GH (n=10, 0.2U/kg rhGH, sc., three times per week for 4 weeks). The immunoglobulin, complement fraction, lymphocyte subsets, and interleukins were determined. Results After treatment, the abnormal elevation of IgG, IgM, C_3, C_4, CD8~+ and CD19~+ in rhGH group decreased significantly at 1 week to 3 weeks; moreover, the abnormal descend of IgA, CD3~+, CD4~+, CD4~+/CD8~+, CD3~+/HLA-DR~+, and CD3~+/CD~ (16+56) in rh-GH group increased significantly at 1 week to 2 weeks compared to those of conventional group; and all recovered to the levels of control at 4 weeks. The interleukin-6 and tumor necrosis factor-? levels in the plasma and supernatant of peripheral blood mononuclearcytes decreased gradually at 1 week to 2 weeks after treatment compared to those of conventional group, and also recovered to the levels of control at 4 weeks. Conclusion rh-GH therapy could significantly improve immune function of children with TOF after radical operation.

BACKGROUND:Current researches on whether the intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells (UC-MSCs) is safe or not lack theoretical basis. OBJECTIVE:To explore the effects of intramuscular injection of heterogeneous UC-MSCs on expression of myocardial vascular endothelial growth factor (VEGF), cardiac troponin I (cTnI) and serum VEGF, hepatocyte growth factor (HGF), insulin-like growth factor 1 (IGF-1) and granulocyte macrophage colony stimulating factor (GM-CSF) in normal Wistar rats. METHODS:A total of 60 Wistar rats were divided into six groups randomly. Rats in the six groups were respectively administrated with intramuscular injection of PBS, Dulbecco’s modified Eagle’s medium, hUC-MSC supernatant, 0.25×105, 1.0×105, 4.0×105 hUC-MSCs. Rats received a second intramuscular injection 4 weeks after first injection. Eight weeks later, the blood sample and myocardial tissue were taken. The serum concentration of VEGF, HGF, IGF-1 and GM-CSF were measured with enzyme-linked immunosorbent assay kit. The myocardial VEGF and cTnI expression were detected by immunohistochemical technique. RESULTS AND CONCLUSION:There was no significant difference in the serum concentration of the VEGF, HGF, IGF-1 and GM-CSF among the groups before and after injection (P>0.05). In the same group, no statistical significant changes in the serum concentration of the VEGF, HGF, IGF-1 and GM-CSF were found among the rats before and after injection (P>0.05). Eight weeks after injection, weak positive expression of the VEGF in the cytoplasm of cardiocytes in the six groups was observed, and strong positive expression of the cTnI in the cytoplasm of cardiocytes in the six groups was observed. There was no significant difference in the VEGF and cTnI content in the myocardium among the groups (P>0.05). Intramuscular injection of hUC-MSCs or the supernatant of hUC-MSCs had no effects on the serum concentration of the VEGF, HGF, IGF-1, GM-CSF and the myocardial VEGF and cTnI expression in normal Wistar rats.

Objective To explore the effects of intramuscular injection human umbilical cord mesenchymal stem cells(hUCMSC)intramuscular injection on the cardiac function and myocardial ultrastructure in rats with Adria-mycin -induced dilated cardiomyopathy (DCM)rats.Methods One hundred and sixty rats were randomly divided in-to a normal group (20 cases)and DCMgroups (1 40 cases),rats in DCMgroups receiving Adriamycin (2 mg/kg)in-traperitoneally once a week for 8 weeks to establish DCM models.The DCM rats were randomly divided into a model control group (served as model group),the supernatant of hUCMSC group (served as supernatant group),the low -dose hUCMSC group(served as low -dose group),the medial -dose hUCMSC group(served as medial -dose group), and the high -dose hUCMSC group(served as high -dose group).Echocardiography was performed to evaluate the car-diac function,plasma brain natriuretic peptide (BNP)level and serum cardiac troponin I (cTnI)level were detected by enzyme -linked immunosorbent assay kit;light microscope and transmission electron microscopy (TEM)were used to observe the ultrastructure of myocardium.Results Rats in the DCM group showed low spirit,declining food intake, progressive emaciation,slow growth,hair loss and ascites.After the intramuscular injection of hUCMSC,the above symp-toms of rats in the low -dose and the medial -dose hUCMSC groups were improved significantly.Before the administra-tion of hUCMSC,the left ventricular ejection fraction (LVEF)[(64.53 ±2.61 )%]and the left ventricular fractional shortening (LVFS)[(30.80 ±2.1 1 )%]were significantly decreased in the DCMgroup compared to those of the con-trol group[(79.67 ±3.02 )%,(43.08 ±3.1 5 )%,all P <0.01 ].After the administration of hUCMSC,LVEF [(75.5 ±7.4)%,(74.0 ±6.1 )%]and LVFS[(40.8 ±3.8)%,(40.2 ±5.0)%]were significantly increased in the low -dose and the medial -dose group compared with those of the model group [(65.8 ±4.5)%,(30.2 ± 2.9)%,all P <0.01 ].The concentration of plasma BNP level [(438.3 ±82.2)ng/L,(341 .7 ±68.9)ng/L]and serum cTnI level [(375.9 ±1 1 0.9)ng/L,(355.9 ±55.6)ng/L]were significantly decreased compared with those of the model group [(449.9 ±91 .8)ng/L,(425.9 ±42.6)ng/L,all P <0.05].The findings of HE staining showed that cardiomyocytes were orderly arranged,edema decreased and cell nucleus homogeneously stained in the low -dose and the medial -dose group.The outcomes of TEM revealed that the ultrastructure of cardiomyocytes was improved in the low -dose and the medial -dose group compared with that of model group,and the cardiomyocyte sarcolemma re-mained intact,and the swelling of mitochondria ameliorated and the cristae of mitochondria remained clear.Conclusions Intramuscular injection of hUCMSC could significantly increase LVEF and LVFS in the Adriamycin -induced DCM rats,and decrease the plasma BNP levels and the serum cTnI levels,attenuate the myocardial pathological damage and improve myocardial ultrastructure.

BACKGROUND:Studies have shown that intramuscular transplantation of xenogeneic umbilical cord mesenchymal stem cel s in a certain dose range is safe and reliable, and it also confirm that this approach is equal y safe and effective for heart failure in rats with dilated cardiomyopathy. OBJECTIVE:To explore the effect of human umbilical cord mesenchymal stem cel s through intramuscular injection on the cytokine expression in adriamycin-induced dilated cardiomyopathy (DCM) rats. METHODS:Total y 160 rats were randomly divided into control group (n=20) and DCM group (n=140). Rats in the DCM group were administered adriamycin intraperitoneal y to establish DCM model. The DCM rats were randomly subdivided into model control group (served as model group), cel supernatant group, the low-dose mesenchymal stem cel group (served as low-dose group), the middle-dose mesenchymal stem cel group (served as middle-dose group), and the high-dose mesenchymal stem cel s group (served as high-dose group). Secondary injection was performed at 4 weeks after first injection. RESULTS AND CONCLUSION:The ELISA test showed that the serum levels of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), leukemia inhibitor factor (LIF) and granulocyte macrophage colony stimulating factor (GM-CSF) were higher in the model group than the control group before and after intramuscular injection (P<0.05). After intramuscular injection, the levels of HGF, LIF, GM-CSF and VEGF in the low-dose group were increased significantly (P<0.05), which were significantly higher than those in the model group (P<0.05). The level of LIF in the middle-dose group was significantly elevated after injection (P<0.05), while there were no significant differences in HGF, VEGF and GM-CSF levels in the high-dose group before and after intramuscular injection (P>0.05). Both the immunohistochemical and RT-PCR results showed that the expressions of insulin-like growth factor-1, VEGF and HGF were increased in al the DCM rats as compared with the control group, which were increased most in the middle-dose group. These findings indicate that low-dose and middle-dose human umbilical cord mesenchymal stem cel s intramuscular injection can increase the serum levels of HGF, LIF, GM-CSF, VEGF and the expressions of IGF-1, HGF and VEGF in the myocardium of DCM rats.

Objective To investigate the immunological pathogenesis of Kawasaki disease( KD)through examination of changes in the expression of suppressors of cytokine signaling 1(SOCS1)and SOCS3,helper T cells and CD4 + CD25 + regulatory T cells(CD4 + CD25 + Treg)in peripheral blood from children with acute KD. Methods Six-teen children[10 boys,6 girls,aged 1 - 2 years old,averaged(1. 6 ± 0. 3)years old]in the acute phase of KD(KD group),16 children[9 boys,7 girls,aged 1 - 3 years old,averaged(1. 5 ± 1. 1)years old]with pneumonia(pneumo-nia group)and 8 normal children[5 boys,3 girls,aged 1 - 5 years old,averaged(2. 0 ± 1. 1)years old]of the same age(normal control group)from the Affiliated Hospital of Qingdao University who were admitted from October 2012 to March 2013 were recruited. The mRNA levels of SOCS1 and SOCS3 in the T cells from peripheral blood were examined by way of reverse transcription - polymerase chain reaction(RT - PCR). Interferon - γ( IFN - γ),interleukin - 4 (IL - 4)and CD4 + CD25 + Treg were quantified by means of fluorescence activated cell sorting(FACS). Results The expressions of SOCS1 and SOCS3,the percentage of IL - 4 T cells observed in the peripheral blood of the pneumonia group were similar to the normal control group(P ﹥ 0. 05),but significantly decreased in the percentage of INF - γ and the level of CD4 + CD25 + Treg(t = 3. 71,12. 81,all P ﹤ 0. 05). Compared to the normal control group and the pneumo-nia group,the expressions of SOCS1 and SOCS3,the percentage of INF - γ and IL - 4 T cells decreased significantly in the peripheral blood of the KD group(t = 2. 27,4. 48,17. 64,2. 73,2. 74,1. 25,2. 36,2. 59,all P ﹤0. 05 ). On the other hand,the level of CD4 + CD25 + Treg in the peripheral blood of the KD group was markedly lower than that in the normal control group(t =7. 70,P ﹤0. 05),but similar to the pneumonia group(P ﹥0. 05). Conclusions The function of helper T cells is inhibited in acute KD. The CD4 + CD25 + Treg may be involved in the immunological pathogenesis of KD.

BACKGROUND:So far, the short-term changes of various organs after injection of umbilical cord mesenchymal stem cells have been reported, but there are few studies on the long-term changes of various organs in healthy rats after repeated intramuscular injection of umbilical cord mesenchymal stem cells. OBJECTIVE:To observe the security of intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells. METHODS:Sixty male SPF Wistar rats were divided into six groups randomly:normal group (suspension liquid of umbilical cord mesenchymal stem cells);control group with culture solution;supernatant group (supernatant of human umbilical cord mesenchymal stem cells);low concentration group (0.25×105 human umbilical cord mesenchymal stem cells);moderate concentration group (1.0×105 human umbilical cord mesenchymal stem cells);high concentration group (4.0×105 human umbilical cord mesenchymal stem cells). Each rat was injected 0.8 mL liquid in muscle, 0.2 mL in each limb, twice at weeks 1 and 4. Biochemical tests were conducted before and after injection. At the end of 8 weeks, al the rats were kil ed and hematoxylin-eosin staining was done with the liver, spleen, lung, kidney, brain and muscle.
RESULTS AND CONCLUSION:There was no abnormal change about biochemical tests and hematoxylin-eosin staining after the intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells. No significant alteration was observed in the liver, spleen, lung, kidney, brain, and muscle of the limb after the injection of heterogeneous umbilical cord mesenchymal stem cells under suitable concentration. These findings indicate intramuscular injection of heterogeneous umbilical cord mesenchymal stem cells at certain concentrations is safe and reliable.

Objective To observe the hindpaw withdrawal threshold (HWT), and the ultrastructure and expression of glia cell line-de-rived neurotrophic factor (GDNF) in sciatic nerve (SN) in chronic constriction injury (CCI) rats after pulsed radiofrequency (PRF). Meth-ods 30 Sprague-Dawley rats were divided into sham modeling-sham treating (SS) group, CCI-Sham treating (CS) group and CCI-PRF (CP) group. The right SNs of the rats in the CS and CP groups were ligated, and it was separated without ligation in the SS group. The CP group accepted PRF at the ligation 14 days after modeling, while the electrodes were placed without electricity in the SS and CS groups. Their HWT was measured before and 1, 7, 14 days after modeling, and 1, 7, 14 days after treatment. The right SN of ligation was observed under electron microscope 14 days after treatment, meanwhile, the GDNF expression was determined with enzyme-linked immunosorbent assay (ELISA). Results HWT was significantly shorter in the CS and CP groups than in the SS group after modeling, and it increased in the CP group 14 days after treatment compared with that of the CS group (P<0.01). The degeneration of SN significantly improved in the CP group compared with the CS group, while the expression of GDNF increased compared with that in the CS and SS groups (P<0.01). Conclusion PRF could relieve the CCI-induced neuropathic pain by upregulating the GDNF expression in the SN to prevent the SN from injury.

BACKGROUND:It is unclear whether intramuscular injection of human umbilical cord mesenchymal stem cells wil increase regeneration of normal myocardial cells and play adverse effects on normal myocardium. OBJECTIVE:To observe the effects of intramuscular injection of human umbilical cord mesenchymal stem cells on expression of the myocardial ki-67, phh3 and cTnT in normal Wistar rats. METHODS:A total of 60 male Wistar rats were divided into six groups randomly:normal group, solution group, supernatant group, low concentration group, moderate concentration group, and high concentration group. These groups were intramuscularly injected different liquid, respectively, as fol ows:PBS, DMEM, the supernatant of human umbilical cord mesenchymal stem cells, human umbilical cord mesenchymal stem cells with amount of 0.25×105, human umbilical cord mesenchymal stem cells with amount of 1.0×105, human umbilical cord mesenchymal stem cells with amount of 4.0×105 . After 4 weeks, each rats received the second same intramuscular injections of human umbilical cord mesenchymal stem cells. Four weeks after the second injection, al rates were kil ed to obtain myocardial tissues which were fixed, embedded and sectioned. Final y, the ki-67, phh3 and cTnT expressions of the myocardium were detected by immunohistochemical technique. RESULTS AND CONCLUSION:In the normal group, negative expression of the ki-67 was observed in the caryon of myocardial cells, weak positive expression of the phh3 was observed in the caryon of myocardial cells, and positive expression of the cTnT was observed in the caryon of myocardial cells. Compared with the normal group, the expression of the ki-67, phh3 and cTnT in the myocardium had no significant difference among the other groups (F=1.076, 0.167, 0.300;P>0.05). Human umbilical cord mesenchymal stem cells or the supernatant of human umbilical cord mesenchymal stem cells via intramuscular injection have no effects on the expression of the ki-67, phh3 and cTnT in normal Wistar rats.

Objective The 3243A ＞ G mutation in mitochondrial DNA is a common cause of the classical mitochondrial diseases characterized by neuro-muscular disorders.This study reports a rare case with the main manifestations of mitochondrial disease in children of mitochondrial cardiomyopathy and respiratory muscle damage.Methods The clinical characteristics,diagnosis and treatment,biochemical,pathological and genetic features of a 10-year-old girl were studied.Results The girl was admitted because of heart failure and respiratory failure at the age of 10.Ragged red fibers in skeletal muscles had been noticed.On her mitochondrial gene,3243A ＞ G mutation,Leu tRNA (UUR),was detected.The mutation rate in the peripheral blood cells was 94％.After the treatment with a high dose of creatine phosphate sodium,coenzyme Q10 and L-carnitine with assisted ventilation,the patient improved rapidly.The child was followed up for 2 years without recurrence.Meanwhile the growth,development and daily life were normal.Conclusions Cardiac and respiratory muscle impairments that appeared at the same time as the first manifestations of the children's mitochondrial disease is not common,and it is rare to have cardiomyopathy based mitochondrial gene 3243A ＞ G mutation is seldom seen clinically.Skeletal muscle biopsy and genetic test is the key for accurate diagnosis.Improving mitochondrial metabolism and assisted ventilation appear to be helpful treatments.