Study on changes of proliferation, migration and invasion abilities of radiation-induced polyploid colon cancer SW1116 cells and their progeny and the related mechanisms / 肿瘤研究与临床

Objective:To investigate the characteristics related to proliferation, migration and invasion of radiation-induced polyploid colon cancer SW1116 cells and their progeny.Methods:Colon cancer SW1116 cells were conventionally cultured in Leibovitz's L-15 medium containing 10% fetal bovine serum. SW1116 cells at logarithmic growth stage were irradiated with 7 Gy X-ray, and the morphological changes of the cells were observed by inverted microscope on days 3, 5, 10 and 19 after radiation induction. According to the morphological changes of the cells, the cells at day 3 after radiation induction were labeled as polyploid giant cancer cell (PGCC) group, and the cells at day 19 were recorded as PGCC progeny group. SW1116 cells without radiation induction were used as control group. Flow cytometry was used to detect cell ploidy in the control, PGCC and PGCC progeny groups, CCK-8 assay was used to detect the proliferation ability of the three groups, cell migration and invasion abilities of the three groups were detected by cell scratch assay and Transwell assay, and Western blotting was used to detect the expressions of cell cycle and proliferation-related proteins and epithelial-mesenchymal transition (EMT) marker N-cadherin (N-cad) in the three groups.Results:The volume of SW1116 cells gradually became larger on days 3, 5 and 10 after radiation induction, and returned to normal on day 19. The proportions of polyploid (DNA content >4N) cell subsets in the control group, PGCC group and PGCC progeny group were (2.3±1.1)%, (23.1±8.1)% and (3.2±0.5)%, the difference was statistically significant ( F = 18.52, P < 0.05), and the proportion of polyploid cell subpopulations in the PGCC group was higher than that in the control group ( t = 5.38, P < 0.01), but the differences between the PGCC progeny group and the control group were not statistically significant ( t = 0.22, P > 0.05). After 72 h of culture, the cell proliferation rates of the control, PGCC and PGCC progeny groups were (100.0±4.1)%, (73.5±0.7)% and (123.9±3.5)%, and the difference was statistically significant ( F = 190.27, P < 0.001). After 48 h of cell scratching, the scratch healing rates in the control, PGCC and PGCC progeny groups were (38.0±2.7)%, (41.5±4.0)% and (63.7±4.2)%, and the difference was statistically significant ( F = 43.05, P < 0.001). After 24 h of culture, the number of invasive cells in the control, PGCC and PGCC progeny groups was 12.9±1.2, 3.4±0.6 and 23.7±1.5, and the difference was statistically significant ( F = 63.64, P < 0.001). The expression levels of cell cycle-related proteins P-cdc25c, cdc25c and cdc2 in the PGCC group were lower than those in the control group (all P < 0.05), and the expression levels of transcription factor-related proteins E2F-2, E2F-3 and EMT marker N-cad were downregulated compared with the control group (all P < 0.05); the expression levels of P-cdc25c, cdc25c, cdc2, E2F-2, E2F-3 and N-cad proteins in the PGCC progeny group were higher than those in the control group (all P < 0.05). Conclusions:Radiation can induce colon cancer SW1116 cells to produce polyploid, which may then generate daughter cells through asymmetric mitosis and gain new life, and then promote the recurrence and metastasis of colon cancer.

Effects of selegiline phosphate combined with liraglutide and osteoporotic triol on glucolipid metabolism and PINP and S-CTX levels in patients with T2DM combined with osteoporosis / 中华内分泌外科杂志

Objective:To investigate the effects of selegiline phosphate combined with liraglutide and osteopontin on glucolipid metabolism, bone mineral density and bone metabolism in patients with type 2 diabetes mellitus (T2DM) combined with osteoporosis (OP) .Methods:A prospective randomized controlled trial was designed.130 patients with T2DM combined with OP admitted to the outpatient clinic of Yantai Yantaishan Hospital from Jan.2020 to Aug. 2021 were selected as study subjects and pregrouped according to the pre-visit serial number according to the random number table method, 65 cases each. The control group weregiven liraglutide and osteopontin combination therapy, and the observation group were treated with sitagliptin phosphate combined with liraglutide and osteopontin, and both groups were treated continuously for 3 months. The tests compared the glycolipid metabolism [glycosylated hemoglobin (HAb1c) , triacylglycerol (TG) , fasting blood glucose (FBG) , total cholesterol (TC) , 2h postprandial glucose (2hPG) , high-density and low-density lipoprotein cholesterol (HDL-C, LDL-C) ], bone density [4 sites of femoral trochanter, lumbar orthostyle, femoral neck, forearm], before and after 3 months of treatment in the 2 groups. Changes in bone metabolism (osteocalcin (OC) , type I collagen C-terminal peptide (S-CTX) , 25-hydroxyvitamin [25 (OH) D], type I procollagen amino-terminal peptidogen (PINP) , and bone-specific alkaline phosphatase (b-ALP) ] levels were detected, and the adverse effects and efficacy of drug administration in the 2 groups were compared.Results:After 3 months of treatment, FBG (7.48±1.02) mmol/L, TG (2.01±0.31) mmol/L, PINP (43.72±4.86) ng/ml, HAb1c (7.43±0.65) %, S-CTX (0.27±0.09) ng/ml, 2hPG (9.08±1.34) mmol/L in the observation group, LDL-C (2.58±0.27) mmol/L were lower than those in the control group (7.86±0.97) mmol/L, (2.29±0.34) mmol/L, (46.55±4.19) ng/ml, (7.81±0.62) %, (0.32±0.10) ng/ml, (10.52±1.41) mmol/L, (2.89±0.31) mmol/L ( t=2.177, 5.968, 3.556, 3.481, 2.996, 5.968, 6.080, P<0.05) ; after 3 months of treatment, the bone density of each site in the observation group was (0.76±0.09) g/cm 3, (0.75±0.10) g/cm 3, (0.76±0.11) g/cm 3, (0.75±0.09) g/cm 3 and OC (20.87±2.33) μg/L, b-ALP (19.70±2.35) U/L were higher than those of the control group (0.70±0.10) g/cm 3, (0.68±0.09) g/cm 3, (0.69±0.10) g/cm 3, (0.70±0.10) g/cm 3, (18.45±3.66) μg/L, (18.09±2.14) U/L ( t=3.596, 4.195, 3.796, 2.996, 4.497, 4.084, P<0.05) ; the overall efficiency of the observation group was 96.92%, higher than that of the control group 84.61% ( χ2=5.876, P<0.05) . The difference was not statistically significant when comparing the incidence of adverse reactions in the 2 groups ( χ2=0.000, P>0.05) . Conclusion:Sitagliptin phosphate combined with liraglutide and osteopontin can further improve clinical efficacy and regulate glucolipid metabolism in patients with T2DM combined with OP, and it also has an important role in improving bone density and regulating bone metabolism level in patients.

Purification and characterization of a sarcosine oxidase from Bacillus sp. BSD-8 / 生物工程学报

We purified a sarcosine oxidase from Bacillus sp. strain BSD-8 isolated from soil. We purified the enzyme by ammonium sulfate precipitation, DEAE-cellulose, Toyopearl hydrophobic and Sephadex G-75 molecular sieve chromatography and characterized the purified sarcosine oxidase. This sarcosine oxidase was a flavin enzyme containing a noncovalently bound flavin with the subunit molecular mass of 51 kDa. The optimal temperature for this enzyme was 60 degrees C and it showed its highest activity at pH 8.5. It was stable in the pH range of 8.0-10.0 and at the temperature of 60 degrees C. Estimated by Lineveaver-Burk plots, the K(m) of the enzyme was 3.1 mmol/L. Ag+, Hg2+, SDS and Tween 80 dramatically inhibted the enzyme activity, whereas Tween 20 and Triton X-100 had no effect on enzyme activity. The thermostability of this enzyme was better than reported sarcosine oxidases, and it could be applied in enzymatic measuring of creatinine.

Evaluation of rheumatoid factor, anti-cyclic citrullinated peptide antibody and anti-keratin antibody in patients with rheumatoid arthritis / 中华风湿病学杂志

Objective To evaluate the role of rheumatoid factor (RF), anti-cyclic citrullinated peptide antibody (ACCP) and anti-keratin antibody (AKA) in patients with rheumatoid arthritis (RA).Methods Serum levels of RF, ACCP and AKA were examined in 82 RA and 56 non-RA patients and their sensitivity and specificity for the diagnosis of RA were exmined. Statistical analysis was performed to test the association between ACCP/AKA and number of tender joints and swollen joints, ESR, CRP, disease activity score (DAS) or Ritchie's articular index (RAI). Results ROC curve was performed for each single auto-antibody and various combinations of any two of the above antibodies. The area under the ROC curve was over 0.5 (P<0.05). The specificity of ACCP and AKA was 91.1% and 92.9% respectively. RF, ACCP and AKA were all sensitive markers for the diagnosis of RA and the sensitivity could be as high as 95.1% when one of these markers was positive. There were statistical differences in the number of swollen joints, ESR and DAS between ACCP positive group and ACCP negative group (P<0.05) and statistical significant difference was observed in tender joint count, swollen joint count, ESR and DAS between AKA positive and negative groups (P<0.05). Conclusion Combined test of ACCP, RF and AKA is useful in routine RA diagnosis.ACCP and AKA may be clinical markers for predicting disease activity and prognosis.