Objective To evaluate the effects and mechanisms of frozen-thawed platelet-rich plasma(PRP)stored at-80 ℃ on wound healing-related cells.Furthermore,to explore the feasibility and effectiveness of using cryopre-served PRP at low temperatures for promoting wound healing.Methods Using non-activated fresh PRP,calcium-activated fresh PRP,and post-thawed PRP,co-cultured with macrophages,fibroblasts,and vascular endothelial cells,the study measured and compared the expression of polarization and inflammatory factors associated with macropha-ges,as well as cell migration and proliferation rates,among other indicators,to analyze the effects of PRP on macro-phage polarization,inflammation,and cell proliferation.Results Compared with the control group,post-cryopre-served PRP at ultra-low temperatures resulted in decreased expression of M1-like polarized macrophage gene iNOS(P＜0.000 1),reduced NO secretion(P＜0.001),increased urea content(P＜0.000 1),decreased M1-related inflammatory factor tumor necrosis factor-alpha(TNF-α)(P＜0.001),and decreased secretion of white blood cell interleukin(IL)-1(P＜0.001);M2-related anti-inflammatory factor IL-10 secretion levels increased(P＜0.01),and IL-12 secretion increased(P＜0.05)Furthermore,co-culturing with frozen-thawed PRP significantly promoted cell migration and enhanced vascular formation efficiency,surpassing the effects of fresh PRP and being comparable to activated PRP(P＜0.01).Cell viability assays and CCK-8 proliferation experiments also showed a significant in-crease in the proliferation rates of L929 and HUVEC co-cultured with frozen-thawed PRP(P＜0.01).Conclusion After being cryopreserved at-80 ℃,PRP has been proven to significantly enhance cell migration,differentia-tion,and proliferation capabilities,while inhibiting the production of inflammatory factors and promoting M2 polari-zation of macrophages.Therefore,low-temperature cryopreservation can be considered as an effective method for PRP preservation.

Objective:To explore the relationship and underlying mechanism between exosomes derived from doxorubicin-resistant osteosarcoma cells and MDR1 and miRNAs. Methods:MG63 and U2OS cell lines were selected to construct doxorubicin-resistant strains, and the 50% inhibitory concentration (half maximal inhibitory concentration, IC 50) of drug-resistant and sensitive strains was detected by MTT, and fluorescence staining was performed at intervals of 15 min between 15 and 120 min to detect the change of fluorescence intensity. RT-PCR and Western Blot were used to detect the expression levels of MDR1 P-gp to verify the drug resistance of osteosarcoma cells. Exosomes were identified by particle size analysis and Western Bolt detection. The endocytosis of PKH26-labeled exosomes from doxorubicin-resistant cells was observed, and the proliferation level and migration of exosomes from doxorubicin-resistant cells co-cultured with osteosarcoma cells were detected by MTT assay and cell scratch assay. The differential expression levels of miRNAs in osteosarcoma-sensitive and drug-resistant cells were verified by sequencing and bioinformatics analysis and RT-PCR assay. Tumor growth, serum exosome identification and mRNA expression level of miR-21-5p in tumor-bearing nude mice between normal osteosarcoma cell group and drug-resistant group, drug-resistant+normal exosome group, drug-resistant+drug-resistant+drug-resistant exosome group were observed. MDR1 expression level in tumor tissue was detected by RT-PCR, Western Blot and immunohistochemistry. Results:The IC 50 of two adriamycin resistant strains were 2.21 vs. 11.81 μg/ml and 0.93 vs. 11.81 μg/ml, respectively, and the fluorescence intensity decreased faster than that of normal strains. The relative mRNA expression levels of MDR1 in two cell lines were normal 1.12±0.16, 1.02±0.11 and drug-resistant 2.15±0.10, 2.127±0.12, respectively. The relative protein expression of P-gp was normal 0.92±0.11, 0.73±0.10 and drug-resistant 0.46±0.03, 0.30±0.04, the differences were statistically significant ( P<0.05). Drug-resistant exosomes can enter osteosarcoma cells through endocytosis and concentrate in the cytoplasm when co-cultured with normal strains. Osteosarcoma cells were co-cultured with drug-resistant exosomes at 2, 4, 6, and 8 μg/ml adriamycin, respectively. Compared with normal group, the proliferation level in drug-resistant group was significantly increased. Compared with the normal cell group 35.95±3.92, 6.72±3.55 and the normal exosome group 51.22±5.55, 19.31±1.93, the drug-resistant cell group 54.20±9.32, 19.24±2.88 and drug-resistant exosome group 76.40±5.41, 30.26±4.87, all had significantly higher cell mobility, the difference was statistically significant ( P<0.05). Exosome sequencing and biogenic analysis of 10 highly upregated miRNAs to validate mRNA expression differences between normal and drug-resistant strains by RT-PCR, showing a significant increase in miR-21-5p expression level of drug-resistant strains (5.89±0.26 vs. 0.99±0.06; 1.05±0.07 vs. 8.80±0.93, P<0.05), the difference was statistically significant ( P<0.05). In MG63 and U2OS, the normal cell group and drug-resistant cell group, and the normal exosome group and drug-resistant exosome group were compared, the tumor volume and the terminal tumor weight of nude mice were increased to varying degrees. MRNA relative expression levels of miR-21-5p in serum exosomes of nude mice after drug intervention were 0.86±0.07 and 0.86±0.05 in normal cell group, respectively. The values were 1.13±0.12, 1.14±0.12 in drug-resistant cell group, 0.71±0.05, 0.75±0.03 in normal exosome group, and 0.90±0.07, 0.93±0.04 in drug-resistant exosome group. Compared with normal and drug-resistant strains, the expression levels of normal and drug-resistant exosome groups were increased, with statistical significance ( P<0.05). Conclusion:The exosomes of drug-resistant cells in osteosarcoma could enhance the proliferation level and migration ability of cells through intercellular transfer of MDR1 and miRNAs. The expression of MDR1 and miR-21-5p in drug-resistant cells and tumor-forming nude mouse serum and tumor tissues were up-regulated which suggested that it might be involved in regulating the drug resistance process of osteosarcoma.

Objective To revise rumination on sadness scale (RSS) and evaluate the reliability and validity of the Chinese version rumination on sadness scale (RSS-C) in Chinese undergraduates.Methods A total of 1 166 undergraduates from 4 universities in Hunan province completed RSS,ruminative response scale (RRS) and Beck depression inventory-Ⅱ (BDI-Ⅱ).The test-retest was conducted in 111 participants 2 weeks later.Results Item analysis and exploratory factor analysis showed the RSS-C included 11 items,consisting of causal analysis,understanding oneself and one's sadness and uncontrollability of ruminative thinking factors.The scores of each item in the high score group were significantly higher than those in the low group(P＜0.001),and the correlation between each item and the total score was significant(r=0.594-0.719,P＜0.001).The confirmatory factor analysis indicated good fit(x2/df=3.938,GFI =0.957,NFI =0.940,CFI =0.954,TLI =0.930,IFI =0.954,RMSEA =0.070).The internal consistency for RSS-C and three factors ranged from 0.694 to 0.868,and the test-retest reliability ranged from 0.620 to 0.833.The scores on RSS-C and three factors were significantly associated with RRS(r=0.555-0.637,P＜0.01),BDI-Ⅱ (r=0.211-0.403,P＜0.01) respectively.Conclusion RSS-C has good reliability and validity and can be used as an effective instrument to assess rumination on sadness in Chinese undergraduates.

Objective To evaluate the reliability and validity of the Chinese-version of the scale for the assessment of illness behavior(SAIB) in Chinese medical college students. Methods In view of the professional and cultural basis of the translation of SAIB,723 medical students were asked to complete the SAIB and the Illness Attitude Scale. After two weeks,75 students were followed to complete the SAIB. Re-sults After the analysis of the scale project analysis and the exploratory factor analysis,23 entries were re-tained and five common factors were extracted( verification of diagnosis,expression of symptoms,medication/treatment,consequences of illness,somatic symptom scanning),and accounted for 56.14％ of the total vari-ance.The confirmatory factor analysis identified a five factors model(SRMR=0.057,χ2/df=1.80,SRMR=0.06,RMSEA=0.05,GFI=0.912,CFI=0.93,TLI=0.91,IFI=0.93,PGFI=0.72,AGFI=0.89).The correla-tion validity between SAIB scale total score and treatment experience and symptoms affected in IAS scale were -0.31,-0.36 (P<0.01).Reliability coefficients of the five dimensions and total score ranged from 0.61 to 0.88,and the test-retest reliability was 0.78. Conclusion The Chinese version of the scale for the assess-ment of illness behavior is a reliable and valid instrument in Chinese.

Objective To explore the reliability and validity of the preliminary revision of the Philadelphia Mindfulness Scale (PHLMS) in Chinese undergraduates.Methods Considering the differences in the cultural and linguistic,the PHLMS was translated and used to conduct a questionnaire survey among 1 221 undergraduates.All participants were also surveyed with five facets mindfulness questionnaire (FFMQ),the white bear suppression inventory (WBSI) and acceptance and action questionnaire-2rd edition (AAQ-Ⅱ) as conduct reliability and validity test.Results The exploratory factor analysis showed that PHLMS could extract two factors,and the cumulative variance contribution rate was 40.355％.The confirmatory factor analysis indicated good fit (x2/df=2.781,CFI =0.945,NFI =0.884,IFI =0.922,TLI =0.906,CFI =0.922,RMSEA =0.053).The internal consistency reliability of the awareness subscales was 0.785,and the internal consistency reliability of the acceptance subscales was 0.772.The correlation coefficients among the PHLMS and FFMQ was significant(r=0.398,P＜0.01).The acceptance subscales were significantly negatively correlated with WBSI and AAQ-Ⅱ(r=-0.458,-0.288,P＜0.01).Conclusion The validity and reliability of the Chinese version of PHLMS meet the measurement criterion.

OBJECTIVE:To study the protective effect of fingolimod on renal ischemia reperfusion injury (RIRI) model mice and its mechanism.METHODS:A total of 60 mice were randomly divided into sham operation group,model group,fingolimod group (1 mg/kg) and fingolimod+wortmannin group [fingolimod 1 mg/kg+phosphatidylinositol 3-kinase (PI3K) specific blocker wortmarmin 1.4 mg/kg],with 15 mice in each group.Except for sham operation group,RIRI model was induced in other 3 groups,and those model mice were given relevant medicine via caudal vein at once 24 h before surgery.Serum of mice were collected in each group after 24 h perfusion.Serum levels of Scr and BUN were measured by automatic biochemical analyzer.The pathological changes of renal tissue were observed under light microscope.The protein expression of intercellular cell adhesion molecule-1 (ICAM-1),monocyte chemoattractant protein-1 (MCP-1) and phosphorylated protein kinase B (p-Akt) in renal tissue were measured by Western blot assay.RESULTS:Compared with sham operation group,the serum levels of Scr and BUN in model group were increased significantly (P＜0.01).Pathological changes were found in the kidney,and RIRI led to widespread renal tubular epithelial cell injury,apoptosis and inflammatory cells infiltration.The protein expression of ICAM-1 and MCP-1 in renal tissue were increased significantly (P＜0.01),the protein expression of p-Akt was increased slightly (P＞0.05).Compared with model group,other indexes of fingolimod group were improved significantly (P＜0.01) except that the protein expression of p-Akt in renal tissue was increased significantly (P＜0.01).Compared with fingolimod group,above indexes of fingolimod+wortmannin group were reversed (P＜0.05 or P＜0.01).CONCLUSIONS:Fingolimod can obviously ameliorate renal injury induced by RIRI in mice,the mechanism of which may be associated with the activation of PI3K/Akt signaling pathway.

Objective To evaluate the clinical efficacy of Qilinxintongshu pill for treatment of critical patients with unstable angina pectoris (UAP) accompanied by upper alimentary tract damage.Methods A prospective randomized controlled trial was conducted. A total of 60 critical outpatients or inpatients with confirmed diagnosis of UAP accompanied by upper alimentary tract damage in Cardiology Department of Hospital of Traditional Chinese Medicine of Qionghai City from November 2011 to January 2015 were enrolled in the study, and they were assigned to a therapy group (30 cases) and a control group (30 cases) by table of random number. The conventional medical basic therapy was given to both groups, in addition, the patients in therapy group took Qilinxintongshu pill (made of notoginseng, dragon's blood, fruit of immature citron, etc.) 5 g once, three times a day, and the patients in control group orally administered clopidogrel 75 mg once a day, the therapeutic course being consecutive 30 days. After treatment, the rates of main cardiovascular events [including death, a newly-happened myocardial infarction (MI), and obstinate ischemia] and events of alimentary damage were compared between the two groups, and adverse effects were observed.Results There was no statistically significant difference in incidence of cardiovascular events between the therapy group and control group [6.7% (2/30) vs. 16.7% (5/30),P > 0.05]. The incidence of massive hemorrhage of gastrointestinal tract was significantly lower in therapy group than that in control group [10.0% (3/30) vs. 36.7% (11/30),P < 0.05]. In the treatment process, there was no significant untoward side effect, and no abnormalities in routine blood and urine tests, liver and renal functions were found.Conclusion Qilinxintongshu pill for treatment of critical patients with UAP accompanied by upper alimentary damage is safe and effective, and does not enhance the incidence of massive hemorrhage of gastrointestinal tract.

Objective To observe the therapeutic effect of Qilin Xintongshu pill for treatment of patients with non-ST segment elevation acute coronary syndromes(NSTE-ACS)accompanied by gastrointestinal bleeding (GIB). Methods A prospective randomized controlled trial was conducted. A total of 67 hospitalized patients in Cardiovascular Department of Qionghai City Hospital of Traditional Chinese Medicine were divided into therapy group(35 cases)and control group(32 cases). Both groups were given conventional medical treatment(except anti-coagulative agent). In the therapy group,the patients received Qilin Xintongshu pill(the ingredients consisting of panax pseudo-ginsen,dragon's blood,immature orange fruit,etc.)oral administration,once 5 mg and 3 times a day,while in the control group,the patients accepted clopidogrel orally,once 75 g and once a day,the therapeutic course in both groups being 30 days. The major cardiovascular events(such as death,a newly-happened myocardial infarction(MI),MI secondary,obstinately ischemia)and incidence of massive hemorrhage of gastrointestinal tract were compared between the two groups,and the adverse events were observed. Results After treatment,the incidence of cardiovascular events and massive hemorrhage of gastrointestinal tract in the therapy group were decreased significantly compared to those in the control group 〔incidence of cardiovascular events:8.57%(3/35)vs. 28.13%(9/32), incidence of massive hemorrhage of gastrointestinal tract:2.86%(1/35)vs. 21.88%(7/32),both P<0.05〕. Adverse events were not observed in both groups,and the examinations of blood,urine,liver and renal functions were of no abnormalities before and after treatment. Conclusion Qilin Xintongshu pill can effectively reduce the incidences of cardiovascular events and massive hemorrhage of gastrointestinal tract in patients with NSTE-ACS accompanied by GIB in the therapeutic course of 30 days,therefore this traditional Chinese herbal medicine is an ideal agent for treatment of such disease.

Objective To evaluate the value of 18F-FDG PET/CT in assessing radiosensitivity enhancement by irisquinone (IR) on rabbit xenografted VX2 lung tumor models.Methods Twenty-four tumor-beating rabbits were randomly divided into 3 groups (8 rabbits/group):group A with radiotherapy alone,group B with combined radiotherapy and IR,and group C without radiotherapy (the control group).18F-FDG PET/CT imaging was performed before radiotherapy and 24 h and one week after radiotherapy.The tumor SUVmax on delayed imaging was calculated in all rabbits.Two rabbits in each group were sacrificed after PET/CT imaging.HE staining was used to assess the differences in cancer cells among groups.Paired t test,one-way analysis of variance and Kaplan-Meier analysis were performed to analyze the data using SPSS 13.0.Results Before radiotherapy,the tumor SUVmax of all the 24 rabbits on standard and delayed imaging were 2.200 ± 0.761 and 3.162 ± 0.833 (t =-5.582,P ＜ 0.01).At 24 h post-radiotherapy,the delayed SUVmax of groups A,B and C were 2.614 ± 0.654,2.349 ± 0.869 and 5.663 ± 1.144,respectively.The differences between pre-radiotherapy and 24 h post-radiotherapy were statistically significant in all three groups (t =2.527,3.620,11.011,all P ＜0.05).One week after radiotherapy,the delayed SUVmax of groups A,B and C were 3.625 ± 1.064,3.058 ±0.850 and 7.424 ± 1.751,respectively.The differences among groups A,B and C were statistically significant (tA∶ B =2.652,tA∶C =3.799,tB∶C =4.366,all P ＜0.05).The cancer cells of group B were fewer than those of groups A and C by pathological findings,which was consistent with 18F-FDG PET/CT results.The survival times of groups A,B and C were (62.375 ±4.534),(69.000 ±4.660) and (54.125 ±5.276) d,respectively.Kaplan-Meier survival curves revealed better survival of group B as compared to groups A and C,respectively (Log-rank,x2 =7.355,16.943,both P ＜ 0.01).Conclusion 18 F-FDG PET/CT is able to evaluate the effect of irisquinone on tumor radiosensitivity enhancement.

Objective To discuss influence of early rehabilitation nursing on rehabilitation effect of patients after heart valve replacement.Methods 60 patients with heart valve replacement were divided into the rehabilitation group and the control group with 30 patients in each group.The control group was given normal nursing and self-exercise ad libitum.Based upon the above,the rehabilitation group was given systemic and normative rehabilitation nursing according to programs established for each individual.The incidence of complications,guardianship time and hospitalization time were compared between the two groups.Results The incidence of complications of the patients in the rehabilitation group was much lower than the patients in the control group.And the time of the patients in the rehabilitation group in ICU and hospitalization time was shorter than the patients in the control group.Conclusions Early rehabilitation care can effectively reduce the incidence of complications after cardiac surgery,promote recovery of cardiac function,also can improve the quality of life of patients.