Objective To investigate the correlations of apparent diffusion coefficient (ADC) with fibrosis and fibroblast activation protein (FAP) score in pancreatic cancer .Methods Eighteen patients with pathologically confirmed pancreatic cancer were per‐formed conventional MR imaging ,DWI examinations .ADCs were measured with region of interest method on a “Single slice” .The wax blocks of 18 patients with pancreatic cancer were received Masson staining and FAP immunohistochemical staining .The correla‐tions of ADC with levels of fibrosis and FAP scores of pancreatic cancer were assessed by Pearson correlation analysis .Results The mild negative correlation between ADC value of cancerous foci and fibrosis was not significant (r= -0 .459 ,P=0 .056) .Significant negative correlation was found between ADC values of moderate and high differentiation cancerous foci and fibrosis (r= -0 .564 ,P=0 .044) .Significant negative correlation was found between ADC value and FAP score (r= -0 .497 , P=0 .036) .Conclusion The negative correlations are found between ADC and fibrosis ,FAP score of pancreatic cancer .DWI will be helpful to infer the pathologi‐cal characteristics .

Purpose: This study aimed to identify the altered pathways and genes associated with freezing damage in human sperm during cryopreservation by multiomics analysis. Materials and Methods: Fifteen fresh human semen samples were collected for transcriptomic analysis, and another 5 fresh human semen samples were obtained for metabolomic analysis. For each semen sample, 1 mL was cryopreserved, and another 1 mL was left untreated for paired design. The results were then combined with previously published proteomic results to identify key genes/pathways. Results: Cryopreservation significantly reduced sperm motility and mitochondrial structure. Transcriptomic analysis revealed altered mitochondrial function, including changes in tRNA-methyltransferase activity and adenosine tri-phosphate/adenosine di-phosphate transmembrane transporter activity. Metabolomic analysis showed that the citrate cycle in mitochondria was significantly altered. Combining transcriptomic, proteomic, and metabolomic analyses revealed 346 genes that were altered in at least two omics analyses. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that metabolic pathways were significantly altered and strongly associated with mitochondria. Five genes were altered in all three omics analyses: COL11A1, COL18A1, LPCAT3, NME1, and NNT. Conclusions Five genes were identified by multiomics analysis in human cryopreserved sperm. These genes might have specific functions in cryopreservation. Explorations of the functions of these genes will be helpful for sperm cryopreservation and sperm motility improvement or even for reproduction in the future.

Objective To explore the influence of triple anti-tumor therapy which bases on sirolimus combined huaier granule and thymosin α-1 on T lymphocyte of rat model with liver cancer recurrence after transplantation.Methods Seventy-two Sprague-Dawley(SD)rats were randomly divided into triple therapy group,sirolimus group,huaier-granule group,thymosin α-1 group,positive-control group and blank group (n=1 2 in each group).Except the blank group,rats in all the other groups were established the simulation animal model of liver cancer recurrence after liver transplantation by chemical-induced method.After the model was established,rats in the positive control group were executed to appraise whether the model was successful.The proportion of regulatory T cells (Treg)of CD4 + T lymphocytes in peripheral blood (Treg%),the percentage of CD4 + T lymphocyte of total lymphocyte(CD4 +T%)and the percentage of CD8 + T lymphocyte of total lymphocyte (CD8 +T%),were detected by the flow cytometry respectively.The relationship between Treg% and CD4 + T %,CD8 + T %,the ratio of CD4 +/CD8 + T lymphocytes(CD4 +/CD8 +)was analyzed by the method of Spearman rank correlation.Results Pathological section of rat liver tissue suggested that the rat model was established successfully.Treg % of positive control group was higher than that of blank group,the difference had statistical significance(P <0.05).Treg% of triple therapy group was significantly lower than that of the positive control group,huaier-granule group,thymosin α-1 group,and significantly higher than the blank group (all in P <0.05 ).Compared with positive-control group,CD4 +T% and CD8 +T% of triple therapy group,sirolimus group and thymosin α-1 group were significantly higher (all in P <0.05).CD4 +T%and CD8 +T% of triple therapy group were significantly higher than those of thymosin α-1 group,sirolimus group and huaier-granule group(all in P <0.05).The relationship between Treg% and CD4 +T%,CD8 +T%, CD4 +/CD8 + in peripheral blood were negatively correlated for rats in each group.In addition,the triple anti-tumor therapy decreased the negative correlation between Treg% and CD4 +/CD8 +.Conclusions Sirolimus based triple anti-tumor therapy can decrease the peripheral blood Treg level of the liver cancer rat,increase the number of T lymphocyte and CD4 +/CD8 +,and play the role of anti tumor cell growth and proliferation.

OBJECTIVETo investigate the effects of mouse adipose-derived stem cell conditioned medium (ADSC-CM) on apoptosis of keratinocytes (human epithelial cell line HaCaT) induced by thermal injury in vitro.

METHODS(1) Adipose-derived stem cells (ADSCs) from inguinal adipose tissue of 5 healthy BALB/c mice were isolated, cultured, and purified by collagenase digestion in vitro. The 3rd passage of cells were collected for morphologic observation, detection of expressions of surface markers CD31, CD34, CD45, CD90, and CD105 with flow cytometer, and identification of adipogenic and osteogenic differentiation. (2) HaCaT cells were incubated in water at 51.5 °C for 35 seconds to reproduce thermal injury model, and then the apoptosis rate was detected immediately after injury by flow cytometer. (3) Thermally injured HaCaT cells were divided into routine culture group (RC, cultured with DMEM containing 10% FBS), serum-free group (cultured with serum-free DMEM), 50%ADSC-CM group (cultured with DMEM containing 50%ADSC-CM), and 100%ADSC-CM group (cultured with 100%ADSC-CM) according to the random number table. After 24 hours, apoptosis of HaCaT cells was observed by acridine orange-ethidium bromide (AO-EB) staining; apoptotic rate was determined by flow cytometer; the mRNA and protein levels of Bcl-2 and caspase-3 were respectively determined by real-time fluorescent quantitative RT-PCR technique and Western blotting (protein level was denoted as gray value); the cell cycles were determined by flow cytometer. All above experiments were repeated for 3 times. Data were processed with one-way analysis of variance and LSD- t test.

RESULTS(1) The 3rd passage of cells proliferated well showing fusiform shape similar to fibroblasts. The positive expression rates of CD31, CD34, and CD45 were less than 10.0%, while those of CD90 and CD105 were above 90.0%. The cells could differentiate into adipocytes and osteoblasts. They were identified as ADSCs. (2) Immediately after injury, apoptotic rate of HaCaT cell was (9.8 ± 0.4)%. (3) The number of apoptotic cells was significantly higher in serum-free group than in the other three groups with AO-EB staining. The apoptotic rate of serum-free group [(8.1 ± 1.2)%] was significantly higher than that of 50%ADSC-CM group [(6.0 ± 0.8)%], group RC [(4.6 ± 0.8)%], or 100%ADSC-CM group [(3.1 ± 0.4)%], with t values respectively 3.02, 4.96, 6.60, P values below 0.01. There was no statistically significant difference in apoptotic rate between group RC and 100% ADSC-CM group (t = 1.50, P > 0.05), while statistically significant difference was found between 100% ADSC-CM group and 50%ADSC-CM group (t = 10.21, P < 0.01). (4) The mRNA level of Bcl-2 of serum-free group (0.34 ± 0.08) was significantly lower than that of group RC, 50%ADSC-CM group, and 100%ADSC-CM group (0.98 ± 0.04, 0.77 ± 0.05, 1.06 ± 0.04, with t values respectively 12.87, 8.07, 14.11, P values below 0.01). Compared with that of 100%ADSC-CM group, the mRNA level of Bcl-2 of group RC was slightly decreased (t = 0.08, P > 0.05) and that of 50%ADSC-CM group was significantly decreased (t = 8.08, P < 0.01). (5) The mRNA level of caspase-3 of serum-free group (1.15 ± 0.05) was obviously higher than that of 50%ADSC-CM group (0.72 ± 0.11), group RC (0.41 ± 0.03), or 100%ADSC-CM group (0.38 ± 0.11), with t values respectively 6.93, 13.97, 22.79, P values below 0.01. Compared with 100%ADSC-CM group, the mRNA level of caspase-3 was slightly increased in group RC (t = 0.05, P > 0.05) and significantly increased in 50%ADSC-CM group (t = 4.77, P < 0.01). (6) The protein level of Bcl-2 was significantly lower in serum-free group (0.93 ± 0.04) than in group RC, 50%ADSC-CM group, and 100%ADSC-CM group (1.74 ± 0.06, 1.32 ± 0.05, 1.90 ± 0.04, with t values respectively 20.45, 11.15, 31.38, P values below 0.01). Compared with that of 100%ADSC-CM group, the protein level of Bcl-2 of group RC was slightly decreased (t = 1.33, P > 0.05), but that of 50%ADSC-CM group was obviously decreased (t = 17.30, P < 0.01). (7) The protein level of caspase-3 was obviously higher in serum-free group (0.63 ± 0.08) than in 50%ADSC-CM group, group RC, and 100%ADSC-CM group (0.46 ± 0.03, 0.29 ± 0.08, 0.21 ± 0.03, with t values respectively 3.28, 5.05, 8.46, P values below 0.01). Compared with that of 100%ADSC-CM group, the protein level of caspase-3 of group RC was slightly increased (t = 0.08, P > 0.05), but that of 50%ADSC-CM group was significantly increased (t = 3.52, P < 0.05). (8) Compared with that of serum-free group, the percentage of cells in G2/M phase of each of the other 3 groups was significantly decreased (with t values respectively 6.88, 4.08, 7.28, P < 0.05 or P < 0.01). Compared with that in serum-free group, the percentage of cells in S phase was significantly increased in group RC and 100% ADSC-CM group (with t values respectively 2.67 and 2.40, P values below 0.05). There was no statistically significant difference in the percentage of cells in G0/G1 phase among all groups (F = 0.70, P > 0.05).

CONCLUSIONS100% xenogeneic ADSC-CM can suppress apoptosis of HaCaT cells induced by thermal injury through regulating the expression of Bcl-2 and caspase-3, and accelerate cell cycle progression by ameliorating the retardation of cell growth in G2/M phase, and all these effects may give rise to some potential in the treatment of burn wounds at early stage.

Adipocytes ; Adipose Tissue ; Animals ; Apoptosis ; physiology ; Burns ; Caspase 3 ; metabolism ; Cell Differentiation ; Cell Line ; Cell Proliferation ; Culture Media, Conditioned ; Fibroblasts ; Humans ; In Vitro Techniques ; Keratinocytes ; metabolism ; physiology ; Mice ; Mice, Inbred BALB C ; Real-Time Polymerase Chain Reaction ; Stem Cells