The effects of epigenetic modification on the differentiation of islet cells and the expression of associated genes (Pdx-1, Pax4, MafA, and Nkx6.1, etc) were investigated. The promoter methylation status of islet differentiation-associated genes (Pdx-1, Pax4, MafA and Nkx6.1), Oct4 and MLH1 genes of mouse embryonic stem cells, NIH3T3 cells and NIT-1 cells were profiled by methylated DNA immunoprecipitation, real-time quantitative PCR (MeDIP-qPCR) techniques. The histone modification status of these genes promoter region in different cell types was also measured by using chromatin immunoprecipitation real-time quantitative PCR methods. The expression of these genes in these cells was detected by using real-time quantitative PCR. The relationship between the epigenetic modification (DNA methylation, H3 acetylation, H3K4m3 and H3K9m3) of these genes and their expression was analyzed. The results showed that: (1) the transcription-initiation-sites of Pdx-1, MafA and Nkx6.1 were highly methylated in NIH3T3 cells; (2) NIH3T3 cells showed a significantly higher level of DNA methylation modification in the transcription-initiation-site of Pdx-1, Pax4, MafA and Nkx6.1 genes than that in mES cells and NIT-1 cells (P<0.05); (3) NIT-1 cells had a significantly higher level of H3K4m3 modification in the transcription-initiation-site of Pdx-1, Pax4, MafA and Nkx6.1 genes than that in mES cells and NIH3T3 cells (P<0.05), with significantly increased level of gene expression; (4) NIH3T3 cell had a significantly higher level of H3K9m3 modification in the transcription-initiation-site of Pdx-1, Pax4, MafA and Nkx6.1 genes than that in mES cells and with NIT-1 cell (P<0.05), with no detectable mRNA expression of these genes. It was concluded that histone modification (H3K4m3 and H3K9m3) and DNA methylation might have an intimate communication between each other in the differentiation process from embryonic stem cells into islet cells.

Objective To investigate the association of serum dehydroepiandrosterone sulfate( DHEA-S) with the cognition in male type 2 diabetes mellitus(T2DM)patients. Methods 99 male patients cared at Tangshan Gongren Hospital and another 97 male healthy controls without T2DM from the medical examination center were recruited. Data on demographic characteristics and clinical parameters were collected, DHEA-S was measured by radioimmunologic assay. Cognitive performance was assessed by the Repeatable Battery for the Assessment of Neuropsychological Status(RBANS). Results (1) Serum DHEA-S levels were lower in male T2DM patients than that of controls[(2. 66 ± 0. 78 vs 4. 02 ± 1. 24) μmol/ L, P<0. 01];(2) Compared with controls, RBANS scores including immediate memory(79. 24 ± 17. 47 vs 86. 25 ± 15. 21, P<0. 01), visuospatial/ constructional(83. 98 ± 17. 98 vs 97. 24 ± 11. 51, P<0. 01), attention(96. 04 ± 14. 65 vs 101. 45 ± 13. 93, P<0. 01), delayed memory (89. 28 ± 11. 74 vs 97. 41 ± 9. 41, P<0. 01), and total scores(85. 85 ± 11. 46 vs 94. 60 ± 10. 91, P<0. 01)were all lower in male T2DM patients;(3) RBANS scores including delayed memory(84. 53 ± 12. 23 vs 93. 94 ± 9. 18, P<0. 01)and total scores(80. 33 ± 10. 91 vs 91. 26 ± 9. 25, P<0. 01)in T2DM patients with low-level DHEA-S(DHEA-S-L)were all lower than those of patients with high-level DHEA-S;(4)Male T2DM patients with cognitive impairment had lower levels of DHEA-S than patients without cognitive impairment(2. 31 ±0. 79 对 2. 90 ±0. 67, P<0. 01);(5) In male T2DM patients, DHEA-S was positively correlated with delayed memory(r = 0. 252, P = 0. 019) and total scores(r=0. 258, P= 0. 016). Conclusion Male T2DM patients are with lower serum DHEA-S levels and worse cognitive performance, and serum DHEA-S was positively correlated with their cognitive performance, suggesting that serum DHEA-S may be involved in the cognitive deficits of male T2DM patients.

Objective:To study the efficacy and success rate of dental pulp revascularization surgery for young permanent teeth with dental pulp necrosis.Method:A total of 100 young patients with dental pulp necrosis of permanent teeth admitted to Xixi Hospital of Hangzhou from March 2019 to March 2023 were selected prospectively, they were divided into the control group and the study group according to the principles of balance and randomization, with 50 cases in each group. The control group and the research group received apical induction shaping surgery and pulp revascularization surgery, respectively. The treatment success rate, root canal wall thickness, crown root ratio, and dental pulp vitality and development of the affected teeth before and after 6 months of treatment were compared between the two groups.Results:The success rate in the study group was higher than that in the control group : 94.00%(47/50) vs. 80.00%(40/50), there was statistical difference ( χ2 = 4.33, P<0.05). After 6 months of treatment, the thickness of the root canal wall in the study group was greater than that in the control group, the affected tooth crown root ratio was lower than that in the control group: (2.43 ± 0.42) mm vs. (2.25 ± 0.39) mm, 0.71 ± 0.09 vs. 0.78 ± 0.11, there were statistical differences ( P<0.05). The proportion of active dental pulp and secondary dental pulp development in the study group during the last follow-up were higher than those in the control group: 92.00%(46/50) vs. 76.00%(38/50), 94.00%(47/50) vs. 80.00%(40/50), there were statistical differences ( P<0.05). Conclusions:The use of dental pulp revascularization surgery in young permanent teeth with dental pulp necrosis can prolong the root of the affected tooth, thicken the root canal ratio, and improve the success rate of treatment by increasing dental pulp vitality and promoting root development.

Objective:To translate the Physical Function Intensive Care Test Score (PFIT-s) into Chinese, and to test its reliability and validity.Methods:Following the translation and back-translation principles of the Brislin model, the Chinese version of PFIT-s was formed by cultural adjustment through the expert consultation. From October to December 2021, convenience sampling was used to select 104 Intensive Care Unit (ICU) patients admitted to Shanghai General Hospital as the research object for investigation to test the reliability and validity of the Chinese version of PFIT-s.Results:The Chinese version of PFIT-s included a total of 4 items, namely assistance (sit to stand) , cadence (steps/min) , shoulder (flexion strength) and knee (extension strength) . A total of one common factor was extracted from exploratory factor analysis, and the cumulative variance contribution rate was 75.97%. The Cronbach's alpha coefficient of the Chinese version of PFIT-s was 0.879, and the scale-level content validity index was 0.958, and the item-level content validity index was 0.833 to 1.000. Using the Chinese version of Intensive Care Units Mobility Scale (IMS) as the criterion, the correlation coefficient between the Chinese version of PFIT-s and IMS was 0.884 ( P<0.01) . Conclusions:The Chinese version of PFIT-s has good reliability and validity and can be used to assess the physical function of ICU patients in China.

Objective:To translate the Physical Function Intensive Care Test Score (PFIT-s) into Chinese, and to test its reliability and validity.Methods:Following the translation and back-translation principles of the Brislin model, the Chinese version of PFIT-s was formed by cultural adjustment through the expert consultation. From October to December 2021, convenience sampling was used to select 104 Intensive Care Unit (ICU) patients admitted to Shanghai General Hospital as the research object for investigation to test the reliability and validity of the Chinese version of PFIT-s.Results:The Chinese version of PFIT-s included a total of 4 items, namely assistance (sit to stand) , cadence (steps/min) , shoulder (flexion strength) and knee (extension strength) . A total of one common factor was extracted from exploratory factor analysis, and the cumulative variance contribution rate was 75.97%. The Cronbach's alpha coefficient of the Chinese version of PFIT-s was 0.879, and the scale-level content validity index was 0.958, and the item-level content validity index was 0.833 to 1.000. Using the Chinese version of Intensive Care Units Mobility Scale (IMS) as the criterion, the correlation coefficient between the Chinese version of PFIT-s and IMS was 0.884 ( P<0.01) . Conclusions:The Chinese version of PFIT-s has good reliability and validity and can be used to assess the physical function of ICU patients in China.

To obtain oleaginous yeast mutants with improved lipid production and growth rates, an atmospheric and room temperature plasma (ARTP) jet was used with a 96-well plate for high throughput screening. Mutants with changes in growth rates and lipid contents were obtained. At a lethality rate of 99%, the positive mutation rate of the yeast cells was 27.2% evaluated by the growth rates of the mutants and the comparison with the wild strain. The fermentation in a medium composed of yeast extract (10 g/L), peptone (10 g/L) and D-glucose (20 g/L) resulted in the lipid yield of the mutant (C4) with 4.07% (W/W) compared with that of the wild strain (1.87%).
Atmosphere ; Biofuels ; Culture Media ; Fermentation ; Glucose ; pharmacology ; Lipids ; biosynthesis ; Mutation ; Peptones ; pharmacology ; Temperature ; Yeasts ; genetics ; growth & development ; metabolism

Objective To investigate the effect of cyclosporine blood level at first year after kidney transplantation on patients with a survival time over 10 years. Methods 380 patients with functional allograft, a survival time over 10 years and long-term administration of cyclosporine A (CsA) were studied, and received CsA-based treatments. According to the blood CsA level at the first year after kidney transplantation, patients were divided into five groups: group 1, blood CsA level was above 0. 208 μmol/L (1 μmol/L = 1201.9 μg/L), group 2, blood CsA level between 0. 166-0. 208μmol/L; group 3, blood CsA blood level between 0. 125-0. 166 μmol/L; group 4, blood CsA blood level between 0. 083-0. 125 μmol/L; group 5, blood CsA level less than 0. 083 μmol/L. Systolic blood pressure (SBP), diastolic blood pressure (DBP), serum creatinine(SCr), uric acid (UA), cholesterol (CH), triglyceride (TG), alanine aminotransferase (ALT), direct bilirubin (DBil) and total bilibubin (TBil), albumin (Alb), hemoglobin (Hb), count of white blood cells and positive rate of proteinuria in 5 groups at the 1st, 5th and 10th year after kidney transplantation were analyzed. Results At the 5th year SBP in groups 1 and 2 was higher than in groups 3, 4 and 5. UA level in group 5 was lower than other groups, and Alb level in group 5 was higher than other 4 groups. Proteinuria positive rate in groups 4 and group was lower than other groups. At the 10th year after kidney transplantation,indexes among 5 groups had no statistically significant difference, except for SBP, DBP, DBil and CH in some groups. There was also no significant difference in SCr level among 5 groups at the 5th or 10th year after transplantation. Conclusion Blood CsA levels at the first year after kidney transplantation has no significant effect on long-term allograft function. But higher level of CsA (＞0. 166μmol/L) at the first year maybe predict high rate of hypertension, high blood UA and proteinuria at the 5th and 10th year after transplantation.

The effects of epigenetic modification on the differentiation of islet cells and the expression of associated genes (Pdx-l,Pax4,MafA,and Nkx6.1,etc) were investigated.The promoter methylation status of islet differentiation-associated genes (Pdx-1,Pax4,MafA and Nkx6.1),Oct4 and MLH1 genes of mouse embryonic stem cells,NIH3T3 cells and NIT-1 cells were profiled by methylated DNA immunoprecipitation,real-time quantitative PCR (MeDIP-qPCR) techniques.The histone modification status of these genes promoter region in different cell types was also measured by using chromatin immunoprecipitation real-time quantitative PCR methods.The expression of these genes in these cells was detected by using real-time quantitative PCR.The relationship between the epigenetic modification (DNA methylation,H3 acetylation,H3K4m3 and H3K9m3) of these genes and their expression was analyzed.The results showed that:(1) the transcription-initiation-sites of Pdx-1,MafA and Nkx6.1 were highly methylated in NIH3T3 cells; (2) NIH3T3 cells showed a significantly higher level of DNA methylation modification in the transcription-initiation-site of Pdx-1,Pax4,MafA and Nkx6.1 genes than that in mES cells and NIT-1 cells (P＜0.05); (3) NIT-1 cells had a significantly higher level of H3K4m3 modification in the transcription-initiation-site of Pdx-1,Pax4,MafA and Nkx6.1 genes than that in mES cells and NIH3T3 cells (P＜0.05),with significantly increased level of gene expression; (4) NIH3T3 cell had a significantly higher level of H3K9m3 modification in the transcription-initiation-site of Pdx-1,Pax4,MafA and Nkx6.1 genes than that in mES cells and with NIT-1 cell (P＜0.05),with no detectable mRNA expression of these genes.It was concluded that histone modification (H3K4m3 and H3K9m3) and DNA methylation might have an intimate communication between each other in the differentiation process from embryonic stem cells into islet cells.

Objective: To evaluate the efficacy and safety of mesenchymal stem cells in allogeneic hematopoietic stem cell transplantation for patients with refractory severe aplastic anemia (R-SAA) . Method: The clinical data of 25 R-SAA patients receiving co-transplantation of mesenchymal stem cells combined with peripheral blood stem cells from sibling donors (10 cases) and unrelated donors (15 cases) from March 2010 to July 2018 in Zhengzhou University Affiliated Tumor Hospital were retrospectively analyzed. Antithymocyte globulin (ATG) treatment was ineffective/relapsed in 11 cases, and cyclosporine (CsA) treatment ineffective/relapsed in 14 cases. Results: There were 13 male and 12 female among these patients. One patient had a primary graft failure, one patient had a poorly engraftment of platelets, and the remaining 23 patients achieved hematopoietic engraftment. The median time of granulocyte engraftment was 12.5 (10-23) days and 15 (11-25) days for megakaryocyte. Incidences of grade Ⅰ/Ⅱ acute graft-versus-host disease (aGVHD) and chronic graft-versus-host disease (cGVHD) were 37.5% (9/24) and 21.7% (5/23) , respectively. There was no severe GVHD and no severe complications that related to transplantation. 21 of 25 (84%) patients were alive with a median follow-up of 22.9 (1.6-107.8) months. The 5-year overall survival rate after transplantation was (83.6±7.5) %. Conclusion The combination of mesenchymal stem cells is reliable and safe in the treatment of R-SAA in peripheral blood stem cell transplantation of unrelated donors and sibling donors, which could significantly reduce the incidence of GVHD and severe transplantation-related complications.

metastatic breast cancer , and analyze the correlation between illness uncertainty and coping style . Methods A total of 98 patients with recurrent or metastatic breast cancer were investigated using Chinese version of Mishel′s uncertainty in illness scale ( MUIS ) and medical coping mode questionnaire ( MCMQ ) . Results The average score of illness uncertainty in patients with recurrent or metastatic breast cancer was (62.50 ±10.89), and the level of illness uncertainty was intermediate .The score of coping style in patients with recurrent or metastatic breast cancer were higher than domestic norm .The scores of avoidance in coping style between two groups was significantly different (P<0.01).Partial correlation analysis indicated that the total score of illness uncertainty was negatively correlated with avoidance in coping style (P<0.05), positively correlated with resignation in coping style (P<0.01) and not correlated with confrontation in coping style (P>0.05).Conclusions Coping style of patients with recurrent or metastatic breast cancer is correlated with their illness uncertainty .Nurses should evaluate illness uncertainty in patients with recurrent or metastatic breast cancer and help them to take effective coping style so as to decrease illness uncertainty and improve therapeutic adherence and confidence in these patients .