Exosomes from human umbilical cord mesenchymal stem cells at passage 2 and 5:a comparative study on microRNA profiles / 中国组织工程研究

BACKGROUND: Exosomes have the function of some mesenchymal stem cells. Understanding the substance composition that plays a representative role in mesenchymal stem cell exosomes will provide clues for further exploration of synthetic exosome analogues. OBJECTIVE: To investigate the difference of microRNA expression profiles in exosomes derived from passage 2 and 5 human umbilical cord mesenchymal stem cells (hUC-MSCs). METHODS: Exosomes in the supernatant of passage 2 and 5 hUC-MSCs were extracted by ultra-high speed centrifugation. The established library was sequenced by using high-throughput sequencing technology. Then we analyzed the sequence results so as to understand the microRNA expression between different groups, and finally did a cluster analysis. RESULTS AND CONCLUSION: 427 657 kinds of microRNAs were detected in the exosomes from passage 2 hUC-MSCs, accounting for 68.93% of the total microRNAs detected; and 119283 microRNAs were detected in the exosomes from passage 5 hUC-MSCs, accounting for 19.22% of the total microRNAs detected. There were 73 526 microRNAs shared between the exosomes from passage 2 and passage 5 hUC-MSCs, accounting for 11.85% of the total microRNAs detected. Bioinformatics analysis (cluster analysis) results showed that these miRNAs were likely to be involved in 161 biological processes, including cell repair, immune and anti-aging. The microRNAs in exosomes from passage 2 to passage 5 hUC-MSCs were largely different. Partial miRNAs exhibited significantly reduced copy numbers. The top five microRNAs with a higher amount, including has-miR-146a-5p, has-miR-191-5p, has-miR-493-3p, has-miR-423-5p, and has-miR-134-5p, have the potential to be the component of synthetic exosome analogues.

A comparative study on serum-free and serum culture methods of human umbilical cord mecenchymal stem cells / 中国组织工程研究

BACKGROUND: Studies have shown increasing risks and problems in the serum culture system, such as immune rejection, batch differences and virus risk. In addition, with the discovery and application of exosomes, the serum-free culture system is becoming an increasing concern. OBJECTIVE: To compare the similarities and differences between the serum-free culture system and the traditional serum culture system, which lays the foundation for the clinical transformation of human umbilical cord mesenchymal stem cells (hUCMSCs) and provides experimental data. METHODS: Umbilical cord was collected from term infants of cesarean section under aseptic condition, and hUCMSCs were isolated and cultured by explant tissue technique. hUCMSCs was cultured with 10% fetal bovine serum (FBS) and 15% serum substitutes (AGS) from the original generation. Then an inverted microscope was used to observe cell morphological changes. Flow cytometry was used to detect cell surface markers. Cell counting kit-8 was used to detect cell proliferation. Induced differentiation experiment was used to detect cell differentiation potential. Western Blot was used to detect the protein levels of oct4, nanog and sox2. RESULTS AND CONCLUSION: Under the inverted microscope, hUCMSCs cultured with AGS showed more uniform vortex-like growth, and those cultured with FBS gradually appeared with cell differentiation or aging with the increase of cell generations. hUCMSCs cultured by both methods expressed CD73,CD90 and CD105 but lowly expressed CD34 and CD45, and there was no significant difference between the two culture methods. FBS method was superior to AGS method in proliferation ability. Results from the induced differentiation experiments showed that hUCMSCs cultured by both methods had adipogenic, osteogenic and chondrogenic abilities, and there was no significant difference between the two culture methods. hUCMSC cultured by both methods expressed oct4 and nanog but showed no significant difference in level, while the expression of sox2 was significantly higher in the hUCMSCs cultured by AGS than by FBS (P < 0.05). To conclude, the hUCMSCs cultured with AGS are in accordance with the international standards of mesenchymal stem cells. The AGS method as an alternative to the FBS method can become a preferred method for hUCMSCs culture.

The impact and mechanism of different doses of yangxinshi on cardiac angiogenesis of acute myocardial infarction in rats / 中国药学杂志

OBJECTIVE: To investigate the influence and mechanism of different doses of Yangxinshi on infarction region angiogenesis of Wistar rats after acute myocardial infarction. METHODS: Sixty healthy male Wistar rats were randomly divided into five groups, named as follow: group A: rosuvastatin group (0.75 mg·kg-1 ·d-1), group B: high-dose Yangxinshi(0.27 g·kg-1 ·d-1), group C: mid-dose Yangxinshi group (0.18 g·kg-1 ·d-1), group D: Low-dose Yangxinshi group (0.09 g·kg-1 ·d-1), group E: saline control group. A, B, C, D group was respectively given the drug by gavage, group E received normal saline by gavage. The models of acute myocardial infarction can be established in the forth week. After continued drugs for 4 weeks, rats were killed before detected blood biochemicalindexes such as blood lipids, liver and kidney function. Myocardial tissue was sliced and stained infarcted myocardium by HE to observe the pathological changes, also extract ischemic and infarct myocardium tissue protein and test VEGF protein expression with immunohistochemistry. RESULTS: Myocardial tissue HE staining were observed a lot of survival island cardiomyocytes and neonatal thin-walled capillaries in four treatment groups, however, control group exist less normal cardiomyocytes and capillaries mainly disappear. Immunohistochemistry results showed high-doses of Yangxinshi group express higher VEGF protein compared with mid-dose group, low-dose group and control group, the difference was statistically significant (P < 0.05), VEGF protein expression was significantly increased the in mid-dose and high-dose Yangxinshi groups than rosuvastatin group, the difference was statistically significant (P < 0.05). CONCLUSION: Yangxinshi promote production of VEGF protein and angiogenesis of ischemic myocardium, in addition its role and its dose is positive correlated. VEGF protein expression was significantly increased the in mid-dose and high-dose Yangxinshi groups than rosuvastatin group, the difference is statistically significant (P < 0.05).

Effects of different dosages of fluvastatin from different dosage forms on glycometabolism in rats with metabolic syndromes / 中国药学杂志

OBJECTIVE: To evaluate the effects of fluvastatin on insulin resistance and hs-CRP level in metabolic syndrome rat model. METHODS: Sixty male KM rats were randomly divided into two groups: 10 rats in control (normal diet) and 50 rats in model (high lipid diet) group. By the end of the 4th week, 10 rats selected randomly from the model group were killed to verify the establishment of metabolic syndrome model. Since the 5th week, the remainder 40 rats were randomly divided into A, B, C and D groups, then were administered intragastrically with fluvastatin capsules at 6 mg · kg-1 · d-1, fluvastatin capsules at 12 mg · kg-1 · d-1, fluvastatin slow-release tablets at 12 mg · kg-1 · d-1 and saline, respectively, and all the rats were fed on normal diet for 4 weeks. All the rats were killed by the end of the 8th week. The levels of TC, TG, LDL-C, HDL-C, FBG, FINS, and hs-CRP were measured respectively and then homeostasis model assessment of insulin resistance was calculated. RESULTS: By the end of the 4th week, the metabolic syndromes model was established. At the end of the experiment, compared with group D, the levels of TC, TG and LDL-C were decreased significantly(P 0.05); compared with group B, the levels of TC, TG, LDL-C, hs-CRP, FBG, FINS and HOMA-IR were decreased significantly in group C (P < 0.05). CONCLUSION: Fluvastatin can reduce TC, TG, LDL-C, FINS, and hs-CRP levels and improve insulin resistance, the effects is more apparent at a large-dosage and with slow-release tablets especially, and that may attribute to its anti-inflammation effects.

Effect of danhong injection on cerebral injury in patients undergoing coronary artery bypass graft operation with extracorporeal circulation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect and its possible mechanisms of Danhong Injection (DHI) on cerebral injury during coronary artery bypass graft (CABG) operation with hypothermic cardiopulmonary bypass (CPB).</p><p><b>METHODS</b>Fifty patients went to CABG with CPB were randomly assigned equally to two groups, the control group and the tested group. DHI 1.5 mL/kg was pumped to where the tested group at the times of aortic pre-charging and unclamping respectively, but to the control group, equal volume of normal saline was given instead. Blood samples were taken from jugular bulb at different time points, i. e. before operation (T1, baseline), re-warming to 36 degrees C (T2 ), 30 min (T3 ) and 6 h (T4) after terminating CPB, for determination levels of superoxide dismutase (SOD) activity using xanthine oxidase method, malondialdehyde (MDA) concentration using thiobarbituric method, tumor necrosis factor alpha (TNF-alpha), as well as interleukin-6, -8 using radioimmunoassay and -10 (IL-6, IL-8 and IL-10) using ELISA.</p><p><b>RESULTS</b>Level of SOD activity significantly decreased during (T2) and after CPB (T3 and T4) in the control group, as compared to the T1 (P < 0.01), but it was unchanged in the tested group; level of MDA increased during and after CPB in both groups (P < 0.01), but more significantly in the control group (P < 0.05), so comparison after CPB between the two groups showed a higher SOD and lower MDA level in the tested group. Plasma levels of TNF-alpha, IL-6, IL-8 and IL-10 significantly increased in both groups after CPB (T3 and T4, P < 0.01) as compared to the T1, but the comparison between groups showed lower plasma TNF-alpha, IL-6 and IL-8 levels at T3 and T4, and higher IL-10 level at T4 in the test group (P < 0.01). All patients had stable life signs with no occurrence of adverse reaction.</p><p><b>CONCLUSION</b>DHI has obvious protective effect on cerebral injury in patients undergoing CABG with CPB, the mechanisms may be associated with the actions of anti-oxidation, anti-inflammation and regulation on immune factors.</p>

Effect of acupuncture on serum insulin level in the patient of simple obesity / 中国针灸

<p><b>OBJECTIVE</b>To compare therapeutic effects of acupuncture plus ear point tapping and pressing therapy and simple medicine for slimming and to probe the mechanism.</p><p><b>METHODS</b>Fifty cases of simple obesity were randomly divided into an acupuncture group and a medication group, 25 cases in each group. The acupuncture group were treated with body acupuncture and electroacupuncture at Tianshu (ST 25), Guanyuan (CV 4), Sanyinjiao (SP 6), Feng-long (ST 40), Zusanli (ST 36), etc., combined with ear point sticking and pressing at Shenmen, Nei-fenmi (endocrine), Pi (spleen), Wei (stomach), Sanjiao (triple energy), Dachang (large intestine), etc.. The medication group were treated with oral administration of Sibutramine, once each day, 10 mg each time. Serum insulin contents before and after treatment were detected, and the therapeutic effect for slimming was assessed.</p><p><b>RESULTS</b>The total effective rate was 88.0% in the acupuncture group and 80.0% in the medication group, with no significant difference between the two groups (P > 0.05); after treatment, the serum insulin levels in the two groups significantly decreased (P < 0.01), and the decrease of insulin level in the acupuncture group was significantly better than that in the medication group (P < 0.05).</p><p><b>CONCLUSION</b>Acupuncture combined with ear point tapping and pressing therapy has a similar therapeutic effect to western medicine for slimming, but the former is better than the later in improving serum insulin level.</p>

The Anti-Inflammation Effect of Atorvastatin on Myocardial Hypertrophy Caused by Pressure Overload / 中华高血压杂志

Background A multitude of studies reported statins prevent the myocardial hypertrophy induced by pressure overload,but the mechanism is unclear.Objective To study the protective effect of atorvastatin on the myocardial hypertrophy caused by pressure overload and it's anti-inflammation effect.Methods Forty Wistar rats were randomized to untreated group(n=10),atorvastatin group(2 mg/kg?d,by gavage,n=10),sham op- eration group(n=10)and control group(n=10).Ventricular hypertrophy model was achieved by surgical con- stricting the abdominal aorta.All rats were sacrificed 8 weeks after the operation and the heart weight to body ratio,left ventricular weight to body ratio,and the size of cardiomyocyte were determined.The expressions of interleukin 18(IL-18)and cardiotrophin 1(CT-1)in the cadiocyte were assessed by reverse transcription polymer- ase chain reaction(RT-PCR).Results Compared with control group,SBP in myocardial hypertrophy group was significantly increased(174?9 vs control 112?15 mmHg,P