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Objective To investigate the effect of exogenous hydrogen sulfide (H2S) on intestinal mucosal barrier after cardiopulmonary resuscitation (CPR) in cardiac arrest (CA) rabbits. Methods Forty-four male New Zealand rabbits were divided into sham operation group (Sham group, n = 12), post-cardiac arrest syndrome (PCAS) group (n = 16) and H2S intervention group (PCAS+NaHS, n = 16) according to random number table method. The rabbit model of PCAS was established by tracheal clamping and suffocation, and CPR was started at 5 minutes after CA. However, Sham group did not clamp the tracheal intubation after anesthesia, and the other operations were the same as those in PCAS group. In the PCAS+NaHS group, a bolus of NaHS (0.5 mg/kg), a H2S donor, was injected via era vein 1 minute before the start of CPR, followed by a continuous injection of NaHS (1.5 mg·kg-1·h-1) for 3 hours, while the rabbits in other group were intravenously injected with the same volume of normal saline (NaCl 0.9%). Intestinal and portal vein blood samples were collected 24 hours after return of spontaneous circulation (ROSC). The level of serum fluorescein isothiocyanate-dextran (FD-4) was detected by fluorescein isothiocyanate (FITC) labeling method to reflect intestinal mucosal permeability. After hematoxylin-eosin (HE) staining of small intestine tissues, the morphological changes of mucosa were observed under light microscope, and the intestinal mucosa injury score was calculated. The expression of tight junction protein ZO-1 in intestinal mucosa was detected by immunohistochemistry. The content of malondialdehyde (MDA) in small intestinal tissue was determined by thiobarbituric acid chromogenic method, the activity of superoxide dismutase (SOD) was determined by xanthine oxidation method, and the level of myeloperoxidase (MPO) was determined by double antibody sandwich enzyme linked immunosorbent assay (ELISA) to reflect the oxidative stress and inflammatory reaction in small intestinal tissue. The expression of apoptosis protein (caspase-3) and autophagy related protein (Beclin-1, LC3) in small intestine tissue was detected by Western Blot. Results 12, 13 and 14 animals were successfully resuscitated in Sham group, PCAS group and PCAS+NaHS group respectively, while 12 animals in each group survived to the end of experiment. Compared with Sham group, the level of FD-4 in portal vein serum was significantly increased in PCAS group (mg/L: 11.95±0.59 vs. 1.43±0.48, P < 0.05), the pathological injury and inflammation infiltration were obviously aggravated under light microscope, the score of small intestine injury was significantly increased (4.21±0.37 vs. 0.36±0.18, P < 0.05), the expression of tight junction protein ZO-1 in the intestine was visibly down-regulated detected by immunohistochemistry, MDA content and MPO activity were significantly increased [MDA (nmol/mg): 3.65±0.32 vs. 1.54±0.24, MPO (U/g): 362±35 vs. 134±18, both P < 0.05], while SOD activity was significantly decreased (U/mg:78.84±7.49 vs. 115.48±8.48, P < 0.05), the expression levels of cleaved capase-3, Beclin-1 and LC3 proteins in the intestine were significantly increased (caspase-3/β-actin: 1.11±0.08 vs. 0.21±0.02, Beclin-1/β-actin: 2.08±0.11 vs. 0.42±0.03, LC3/β-actin: 1.05±0.07 vs. 0.37±0.05, LC3-Ⅱ/ LC3-Ⅰ: 1.28±0.14 vs. 0.17±0.02, all P < 0.05). Compared with PCAS group, the portal vein serum FD-4 level in PCAS+NAHS group was significantly decreased (mg/L:5.59±0.48 vs. 11.95±0.59, P < 0.05), the intestinal mucosal pathological injury and inflammatory cell infiltration were significantly decreased, the score of small intestine injury was significantly decreased (2.18±0.47 vs. 4.21±0.37, P <0.05), the expression of ZO-1 in intestine was significantly increased, MDA content and MPO activity in intestine were significantly decreased [MDA (nmol/mg): 2.65±0.31 vs. 3.65±0.32, MPO (U/g): 251±21 vs. 362±35, both P < 0.05], while SOD activity was significantly increased (U/mg: 96.86±7.52 vs. 78.84±7.49, P < 0.05), while the expression of activated caspase-3, Beclin-1 and LC3 proteins was significantly decreased (caspase-3/β-actin: 0.72±0.06 vs. 1.11±0.08, Beclin-1/β-actin: 0.96±0.08 vs. 2.08±0.11, LC3/β-actin: 0.72±0.06 vs. 1.05±0.07, LC3-Ⅱ/ LC3-Ⅰ:0.83±0.09 vs. 1.28±0.14, all P < 0.05). Conclusion H2S has a protective effect on intestinal mucosal injury induced by CA/CPR, which may be related to tight junction protein ZO-1 up-regulation, oxidative stress alleviation, inflammation reduction, apoptosis and autophagy inhibition.
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Objective To approach the effects of full Marathon on striated muscle and renal function of Marathon amateurs without complaints. Methods A prospective self-paired design study was conducted. The amateurs without body discomfort, hematuria, brown urine, or persistent muscle pain within 1 week after the 2012 Xiamen International Marathon Race were enrolled voluntarily. The peripheral blood and random urine specimens of all subjects under static status 1 week before the race and after the race instantly (within 10 minutes after finishing the race) were collected to detect markers of renal function and striated muscle injury. Results Sixty-one subjects were included in the final analysis of the study with full Marathon of 42.195 km and mean race time of (297.05± 55.60) minutes. Compared with those under static status before the race, the markers of renal function including the levels of urinary N-acetyl-beta-D-glucusamidase [NAG (U/L): 64.00 (54.50, 85.50) vs. 9.50 (8.10, 11.50)], urinary β2-microspheres protein [β2-MG (μg/L): 261.00 (128.50, 1 608.00) vs. 66.60 (33.75, 123.00)], random urinary creatinine [UCr (μmol/L): 19 066.56±10 938.31 vs. 5 872.52±4 363.20] and serum creatinine [SCr (μmol/L): 129.97±25.84 vs. 97.39±14.51] immediately after the race were significantly increased (all P < 0.01); the markers of muscle injury including the levels of serum creatine kinase [CK (U/L): 864.00 (504.00, 1 644.00) vs. 164.00 (128.00, 256.00)], lactic dehydrogenase [LDH (U/L): 383.26±141.69 vs. 182.23±41.12], myoglobin [Mb (mg/L): 1 880.00 (1 080.00, 3 300.00) vs. 42.00 (36.00, 54.50)], alanine aminotransferase [ALT (U/L): 27.0 (19.5, 38.0) vs. 24.0 (15.0, 29.5)] and aspartate transaminase [AST (U/L): 52.07±25.13 vs. 28.28±11.86] were also significantly increased (all P < 0.01), and the increase in CK, Mb, and LDH were more significant. It was shown by correlation analysis that CK after race was negatively correlated with age (r = -0.352, P = 0.005) and body mass index (r = -0.271, P = 0.035), and it was positively correlated with racing time (r = 0.387, P = 0.002) and urinary β2-MG after the race instantly (r = 0.364, P = 0.004). Mb after race was negatively correlated with body mass index (r = -0.331, P = 0.009), and it was positively correlated with urinary β2-MG after the race instantly (r = 0.315, P = 0.013). LDH after race was negatively correlated with age (r = -0.275, P = 0.032) and body mass index (r = -0.377, P = 0.003), and it was positively correlated with urinary β2-MG after the race instantly (r = 0.424, P = 0.001). Conclusion Full Marathon could significantly impact striated muscle and renal function of Marathon amateurs without complaints.
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Objective: To evaluate the quality consistency of the original product ( DIOVAN ) and generic valsartan capsules ( VSC) . Methods:The dissolution of the two capsules in four media was monitored by the UV method described in Chinese Pharmaco-poeia ( ChP 2010 edition) . The f2 similarity factor approach was used to evaluate the dissolution similarity between DIOVAN and VSC and the production quality of the enterprises. Meanwhile, the homogeneity in the same batch was studied by dissolution precision and the reproducibility in the different batches were also evaluated by a similar equivalent limit method to show the production quality of the manufacturers. Results:The dissolution of VSC and Diovan in pH 6. 8 phosphate buffer medium was both above 85% in 15 min. The f2 similarity factors in the other three media ( water, pH 4. 5 acetate buffer and 0. 1 mol·L-1 HCl) were all above 50. The relative standard deviation ( RSD) of precision in the same batch was less than 10%. Among the different batches, dissolution limit value ( Q) was within the range of the upper and lower limit value of probability levels (δ) . Conclusion:The f2 similarity factor results indicate the in vitro dissolution of the reference drug ( DIOVAN) and generic drug ( VSC) is consistent in the four media. The production quali-ty of generic manufacturer is also good with promising homogeneity and reproducibility evaluation results.
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Objective To study whether the viral metabolites can make influence on biological behavior of mesenchymal stem cells (MSCs) and to provide evidence for safe MSCs transplantation.Methods MSCs were isolated from bone marrow of C57BL/6 mice and the mRNA level of Toll like receptor 3 (TLR3)was detected.In vitro polyinosinic-polycytidylic acid [poly(I ∶ C)] was used as the product of viral metabolite to stimulate MSCs.We recorded the change of MSCs in terms of self-renewal,multi-directional differentiation,directional migration and immunosuppression by flow cytometry,PCR etc.Results ①Low-expressed TLR3 mRNA was dectected in MSCs and it was upregulated after being stimulated by poly(I ∶ C).②After being stimulated by poly(I ∶ C)for 24,48,and 72 hours,the self-renewal ability of MSCs had no statistical difference with that of the control group(P ＞0.05).However,poly (I ∶ C)promoted the differentiation of MSCs to lipoblast and osteoblast (P ＜ 0.01).③The directional migration ability of MSCs was weakened significantly by poly(I ∶ C) stimulation.After being stimulated for 6,12,and 24 hours,the migration rate of MSCs was(82.83 ± 1.69) ％,(87.80 ± 3.58) ％,and (92.67 ± 2.52) ％ respectively compared to that of the control group.The difference had statistical significance(P ＜0.01).④MSCs had great immunological suppression ability and it was dependant on the number of MSCs.After being stimulated by poly(I ∶ C),the ability of MSCs' immunological suppression on proliferation of activated splenocytes was not impaired and the difference had no statistical significance compared with that of the control group(P ＞0.05).Conclusion While maintaining the viability and immunosuppression function of MSCs,viral metabolite inhibits migration of MSCs,holds them in the transplanted position and protects the graft,providing evidence for MSCs as seed cells in the treatment of diabetes and other immune disorder diseases.
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Objective To study the protection of mouse bone marrow-derived mesenchymal stem cells ( MSCs) for allogenic islets. Method MSCs from C57BL/6 mice were preceded to a 24-well culture plate with the density of 3 × 104/well. On the second day, islets were isolated, purified and divided to undergo streptozotocin (STZ) induced chemical injury and mixed lymphocyte reaction ( MLR) respectively. Then, treated and control islets were respectively divided into the following groups: islets + MSCs, STZ-islets + MSCs, and MLR-islets + MSCs. As control groups for their counterparts, treated or non-treated islets were also cultured without MSCs. On the 5th day incubation, glucose-stimulated insulin secretion test was performed to assess the function of islets in different groups, comparing their insulin-secretion amount stimulated by low or high glucose and the stimulation index determined by the ratio of (insulin amount secreted under high-glucose stimulation)/(insulin amount secreted under low-glucose stimulation ). Islet viability was evaluated by acridine orange (AO)/propidium iodide (PI) fluorescence staining. Result As shown by AO/PI staining, large numbers of dead cell with red fluorescence could be observed in STZ- or MLR- treated islets without MSCs, while the number of dead cells obviously reduced in MSC-cocultured islets with increased viable cells of green fluorescence. STZ- or MLR- treated islets exhibited apparently decreased insulin-secretion amount either under low- or high-glucose stimulation, as well as the stimulation index. The insulin-secretion function was significantly improved in islets cocultured with allogenic MSCs (P ＜ 0. 05 ). Conclusions Bone marrow-derived MSCs can protect isolated allogenic islets against chemical and immunological injury.