AIM:To investigate the effect of advanced glycation end products on inflammation in cultured cardiomyocytes.METHODS: Primary cardiomyocytes were isolated from Sprague-Dawley neonatal (1 to 2 days old) rats ventricles. The insulin resistant cardiomyocyte model was established. Neonatal rat ventricular myocytes were exposed to AGEs for 24 hours. TNF-? mRNA and PPAR-? mRNA expressions were determined by RT-PCR. Activation of NF-?B in the cells was examined by using immunocytochemistry. The ultrastructure of the cells was detected by transmission electron microscope.RESULTS: The exprssion of TNF-? mRNA and the activation of NF-?B increased, the expression of PPAR-? mRNA decreased compared with control group (P

Objective To investigate the effect of JUC on the incisions for episiotomy.Methods Three hundred primiparas to undergo episiotomy in our hospital were divided into two groups in equal number.The experiment group was given JUC Spray before suturing and the control group did not use any solution.In the two groups,antibiotics were not used after the operation,and the incisions were only cleaned with 0.5%povidone-iodine 2 times a day.Result There were significant differences between the two groups in terms of postoperative pains,inflammation and healing in the wounds,and hospital stay(P<0.05).Conclusions Application of JUC after episiotomy could be long-acting in antibacteria.It can reduce wound pain,improve wound healing rate, decrease the medical cost and shorten the hospital stay.

Objective To establish a mouse model of lymph node metastasis of breast cancer cells by luciferase imaging assay,to monitor early process of lymph node metastasis,and to evaluate the effect of X-ray radiation therapy on tumor.Methods The mouse mammary cancer cell line 4T1-Luc expressing luciferase was inoculated subcutaneously into the paw pad of nude mice to establish a model of subcutaneous lymph node metastasis.The lymph node metastasis in nude mice was continuously observed by in vivo fluorescence imaging system,and the nude mice with early lymph node metastasis of breast cancer cells were divided into control group and treatment group randomly.The radiotherapy effect was observed by in vivo fluorescence imaging system and evaluated by the pathological changes of HE staining of tumor tissue.Results A mouse lymph node metastasis model of breast cancer cells was successfully established,and the volume of primary tumor in paw correlated with the fluorescence photon number positively (r =0.958,P ＜ 0.001).On the twenty-fourth day after inoculation,the fluorescence photon number in pad tumor and popliteal fossa tumor of treatment group were significantly decreased in comparison with the control group (t =32.58,P ＜ 0.05),and the inhibition ratio of radiotherapy on tumor growth approached to 85 ％.HE staining showed that the apoptosis and necrosis in irradiated tumor was obviously higher than that in control group.Conclusions Bioluminescence imaging technique can be used to evaluate the effect of X-ray on breast cancer suppression and lymph node metastasis in mice.

AIM:To investigate radiosensitization effect of apatinib, a vascular endothelial growth factor (VEGF) receptor2 tyrosine kinase inhibitor, on human gastric carcinoma cell line SGC-7901 and its mechanism.METHODS:SGC-7901 cells were divided into control group, apatinib group, radiotherapy group and combination group.The cell viability was measured by CCK-8 assay.The changes of cell apoptosis and cell cycle were analyzed by flow cytometry.The protein levels of cell apoptosis biomarkers, such as PARP, cleaved caspase-9, cleaved caspase-3 and Bcl-2, and cell proliferation biomarkers, p-PLCγ1 and p-ERK1/2, were detected by Western blot.γ-H2AX expression was detected by immunofluorescence.RESULTS:Compared with apatinib group and radiation group, the cell viability was inhibited after treatment with both apatinib and X-ray (P<0.01).The protein levels of cell proliferation markers p-PLCγ1 and p-ERK1/2 were down-regulated.The cell apoptosis was enhanced (P<0.01).The protein levels of cell apoptosis makers such as PARP, cleaved caspase-9 and cleaved caspase-3 were up-regulated, while Bcl-2 was down-regulated.The disappearance of γ-H2AX foci in the nucleus was delayed, indicating that apatinib impaired the repair of radiation-induced DNA double-strand breaks.The proportion of G2 phase was significantly increased (P<0.01).The combination treatment had more significant effect on SGC-7901 cells than treating with apatinib or radiotherapy alone.CONCLUSION:Apatinib increases the radiosensitivity of gastric cancer cells via blocking VEGF pathway.

Objective To develop a new method to expose the stellate ganglion to increase the success rate of establishing a dog model of atrial fibrillation indinced by sympathetic stimulation .Methods A total of 28 adult dogs were randomly divided into traditional group and improvement group , 14 dogs in each group .The stellate ganglions were separated by the two different methods , respectively , to establish a sympathetic stimulation induced atrial fibrillation model in all the dogs .Changes of vital signs , survival rate of the dogs and the voltage required to stimulate the stellate ganglion were recorded intraoperatively .Changes of cardiac electrophysiology were recorded before and after electric stimulation . The levels of released neurotransmitters were detected by immunohistochemistry . Results The survival rate of the improvement group was 100%(14/14), significantly higher than the 64.3%(9/14) of the traditional group (P<0.05). The operation time of the improvement group was 122.71 ±3.62 min, significantly shorter than the 269.44 ±8.79 min of the traditional group (P<0.05).The threshold voltage of the improvement group was significantly lower than that of the traditional group ( P<0.05) .Conclusions Our modified surgical procedure can effectively reduce the mortality of dogs , significantly shorten the operation time , and reduce the intraoperative blood loss , keeping a more intact stellate ganglion , and maintains a more stable voltage of electric stimulation , Therefore, it is a new method more suitable for establishment of a sympathetic stimulation induced atrial fibrillation model in dogs .

This study was aimed to analyze the content variations of chemical constituents produced in fresh Radix Rehmanniae (Xian-Di-Huang) after processing into Radix Rehmanniae Recens (Sheng-Di-Huang) and Radix Rehmanniae Preparata (Shu-Di-Huang). HPLC was used in the study of preparing Xian-Di-Huang into Sheng-Di-Huang, and the processing into Shu-Di-Huang. The contents of two newly produced chemical components, which were 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP) and 5-hydryoxymethyl-furfural (5-HMF). The contents of both chemical components were determined in Sheng-Di-Huang and Shu-Di-Huang bought from the market. The Zorbax SB-C18 column (250 mm í 4.6 mm, 5 μm) was used. The mobile phase was methanol-wa-ter (5:95). The flow rate was 1.0 mL·min-1. The detection wavelength was set at 280 nm. The results showed that no DDMP or 5-HMF was detected in Xian-Di-Huang. However, after processing, DDMP and 5-HMF can be de-tected in both Sheng-Di-Huang and Shu-Di-Huang. The content in Shu-Di-Huang was higher than that in Sheng-Di-Huang. Both contents in Shu-Di-Huang were gradually increased along with processing time. The con-tent reached to the highest level after processing for 24 h and 32 h, respectively. And then, the content decreased. Both DDMP and 5-HMF were detected from three batches of Sheng-Di-Huang and ten batches of Shu-Di-Huang bought from the market. The content of 5-HMF was higher in Shu-Di-Huang than in Sheng-Di-Huang. There was no obvious difference in the content of DDMP between Sheng-Di-Huang and Shu-Di-Huang. It was concluded that DDMP and 5-HMF were produced in the processing Sheng-Di-Huang and Shu-Di-Huang. The contents were gradually increased along with the prolonging of processing time. There was obvious difference in the content of 5-HMF in Shu-Di-Huang and Sheng-Di-Huang.

Objective To explore the effect of dronedaronel on hyperpolarization-activated cyclic-nucleotide-gated(HCN) channel expression by detecting the change of HCN channel mRNA and protein level before and after giving dronedarone in neonatal rat ventricular myocytes.Methods Neonatal rat ventricular myocytes were separated and digested by type Ⅱ collagenase,and then single ventricular myocytes were collected through differential sticking wall separation method.According to the concentrations (0.1,0.5,1.0,5.0,10.0,20.0 μmol/L of dronedaronel for treating myocytes for 48 h) and time(10 μmol/L of dronedaronel for treating myocytes for 1,6,12,24,48 h)the gradient grouping was conducted.The levels of HCN2 and HCN4 channel mRNA and protein level were determined by real-time PCR and Western blot.Results The HCN2 mRNA and HCN4 mRNA expression levels in concentration gradient group and time gradient group were lower than those in the control group(P＜0.05);compared with the control group,the protein level in the 10 umol/L dronedaronel treatment for 12 h group was significantly down-regulated(P＜ 0.01).Conclusion Dronedaronel could inhibit the expression of HCN2/HCN4 channel mRNA and protein,moreover its action shows the concentration dependency and reaches the maximum at 12 h after medication.

OBJECTIVETo explore the inhibitory effect of He-Ne laser irradiation on fibroblast growth of hypertrophic scars in culture.

METHODSHe-Ne laser with wavelength of 632.8 nm, power density of 50 mW/cm(2) and doses of 3 J/cm(2), 30 J/cm(2), 90 J/cm(2) and 180 J/cm(2) was used to irradiate human scar fibroblasts in culture 1, 3 and 5 times respectively, and then the cell count and cell cycle analysis were done.

RESULTSRepeated irradiation with He-Ne laser at dose of 180 J/cm(2) three and five times led to an evident decrease in total cell number compared with that of the control group and there was a significant difference (P<0.05). The cell cycle analysis showed after three and five times of irradiation with 180 J/cm(2) He-Ne laser the cell number in S-phase decreased from 51% to 20% and 14% respectively, the cell number in G(0)/G(1) phase increased from 28% to 55% and 60% respectively, and the cell percentage in Sub-G1 phase was 6.7% and 9.8% respectively.

CONCLUSIONSRepeated irradiation with 180 J/cm(2) He-Ne laser can inhibit scar fibroblasts growth in culture. It may be that He-Ne laser irradiation causes cell stagnation in G(0)/G(1) phase and apoptosis.

Cell Division ; radiation effects ; Cells, Cultured ; Cicatrix ; pathology ; Dose-Response Relationship, Radiation ; Female ; Fibroblasts ; cytology ; radiation effects ; Helium ; Humans ; Lasers ; Male ; Neon

OBJECTIVETo analyze the correlation between mitral regurgitation grading and left ventricular ejection fraction in elderly patients (>60 years of age) in a 2-year follow-up.

METHODSA total of 455 patients with the diagnosis of at least mild mitral regurgitation by echocardiography were divided into ischemic mitral regurgitation (IMR) group and non-ischemic regurgitation (NIMR) group. The patients were followed up with echocardiography every 6 months and the data were analyzed at the end of 24 months.

RESULTSMitral regurgitation grade was inversely correlated with left ventricular ejection fraction (LVEF). Patients with moderate and severe IMR had a lower LVEF than those with NIMR (P<0.05). After adjustment for age, sex, body mass index, high blood pressure, diabetes, atrial fibrillation and cardiomyopathy, the mean LVEF at 2 years was lowered by 2.7% (1.4%-4.1%), 2.7% (1.3%-4.0%), and 5.2% (3.5%-6.9%) in mild, moderate and severe IMR patients, respectively (P<0.04), and by 3.2% (1.6%-4.8%), and 3.0% (1.4%-4.5%), and 1.7%(-0.5%-3.9%) in mild, moderate and severe NIMR patients (P=0.30).

CONCLUSIONThe mean LVEF in IMR patients is significantly lowered compared to that in NIMR patients. The grade of mitral regurgitation is inversely correlated with the regurgitation area in IMR patients. Stratified management might help improve LVEF in severe IMR patients.

Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Male ; Mitral Valve Insufficiency ; Stroke Volume ; Ventricular Dysfunction, Left ; physiopathology

Objective: To investigate the effects of human amniotic epithelial stem cells-derived exosomes on healing of wound with full-thickness skin defect in rats. Methods: (1) Human amniotic epithelial stem cells were isolated from the amnion tissue of 5 full-term pregnant women in Department of Obstetrics of our hospital by the method of trypsin digestion, and their morphology was observed. The third passage of cells were stained with rhodamine-phalloidin for cytoskeleton observation. The third passage of cells were identified with flow cytometry through the detection of expressions of cell surface markers CD29, CD31, CD34, CD90, CD105, SSEA3, SSEA4 and immunity-related marker human leukocyte antigen-D related site (HLA-DR). The third passage of cells were also assessed the ability of adipogenic and osteogenic differentiation. (2) The third passage of human amniotic epithelial stem cells were cultured in DMEM medium supplemented with 10% exosome-free fetal bovine serum. Exosomes were isolated from culture supernatant by the method of ultracentrifugation and represented with scanning electron microscope for morphologic observation. (3) Six adult SD rats were anesthetized, and four 1 cm×1 cm sized wounds with full-thickness skin defect were made on the back of each rat. The wounds on the back of each rat were divided into control group, 25 μg/mL exosomes group, 50 μg/mL exosomes group, and 100 μg/mL exosomes group according to the random number table (with 6 wounds in each group), and a total volume of 100 μL phosphate buffered saline, 25 μg/mL exosomes, 50 μg/mL exosomes, and 100 μg/mL exosomes were evenly injected around the wound through multiple subcutaneous sites, respectively. The wound healing rate was calculated based on measurement on post injury day (PID) 7, 14, and 21. On PID 21, the healed wound tissue of each group was collected and stained with HE to observe and count skin accessories, and the arrangement of collagen fibers was observed with Masson staining. Data were processed with analysis of variance for repeated measurement, analysis of variance of randomized block design, one-way analysis of variance, and Bonferroni test. Results: (1) The cells, which were isolated and cultured, displayed typical cobblestone morphology with many microvilli on cell surface. Among the cells, the positive expression rates of CD29, CD90, SSEA3, and SSEA4 were above 50.0%, and the rate of CD105 was 8.0%, while the rates of CD31, CD34, and HLA-DR were almost 0. The cells could differentiate into adipocytes and osteoblasts. The above results revealed that the cells cultured were human amniotic epithelial stem cells. (2) Human amniotic epithelial stem cells-derived exosomes were round or oval vesicles with diameter from 50 to 150 nm. (3) On PID 7 and 21, wound healing rates of the four groups were close (with P values above 0.05). On PID 14, wound healing rates of 50 and 100 μg/mL exosomes groups were (89.8±4.3)% and (92.0±4.6)% respectively, significantly higher than the wound healing rate of control group [(80.3±6.4)%, P<0.05 or P<0.01]. Moreover, the wound healing rate of 100 μg/mL exosomes group was significantly higher than that of 25 μg/mL exosomes group [(83.3±5.1)%, P<0.05]. On PID 21, the numbers of skin accessories in 50 and 100 μg/mL exosomes groups were 4.3±1.4 and 5.1±1.6 respectively, obviously more than those of control group and 25 μg/mL exosomes group (respectively 1.4±0.5 and 1.8±0.6, with P values below 0.01). Well reorganized collagen fibers were observed just in the healed wound tissue of 50 and 100 μg/mL exosomes groups. Conclusions Human amniotic epithelial stem cells-derived exosomes can promote healing of wound with full-thickness skin defect in rats.