Glioma is difficult to treat due to the unique tumor microenvironment and blood-brain barrier. (13aS)-3-Hydroxyl-6,7-dimethoxyphenanthro[9,10-b] indolizidine (PF403), a phenanthroindolizidine alkaloid, has been identified as a promising therapeutic agent for the treatment of glioma. However, the anti-glioma mechanism of PF403 in vivo has not been conclusively verified and must be further elucidated. Hence, a strategy without chemical modification was applied to identify the target of PF403. In this study, we identified nicotinamide phosphoribosyl transferase (NAMPT) as the target of PF403 by using thermal proteome profiling (TPP). Moreover, microscale thermophoresis (MST), surface plasmon resonance (SPR), and isothermal titration calorimetry (ITC) experiments confirmed that NAMPT exhibits good affinity for PF403. Direct and indirect enzyme activity assays revealed that PF403 inhibited the catalytic activity of NAMPT, leading to a decrease in the concentration of nicotinamide adenine dinucleotide (NAD⁺) in U87 cells. X-ray diffraction and amino acid spot mutation experiments revealed that PF403 primarily relies on the formation of pi-pi interactions with residue Tyr188 to maintain binding with NAMPT (PDB code 8Y55). After NAMPT was knocked down with lentivirus, PF403 lost or partially lost its antitumor activity at the cellular and animal levels. These findings suggest that PF403 exerts antitumor activity by directly targeting NAMPT.

Objective:To investigate the application of four-grade functional activity score (FAS) in early pain management of patients after radical mastectomy, so as to provide theory basis for the early management of cancer pain.Methods:Ninety-six patients underwent radical mastectomy in Sichuan Maternal and Child Health Hospital from April 2019 to June 2020 were enrolled, and divided into two groups by random number table methods, each with 48 cases. The control group received routine nursing combined with Numerical Rating Scale (NRS), based on this, the study group received FAS. Then the pain degree, first activity time of out-of-bed, surgical wound recovery time, drainage tube placement time, hospital stay length, quality of life (FACT-B), upper limb function, and incidence of complications were compared between the two groups.Results:NRS score at the initial event, first activity time of out-of-bed, surgical wound recovery time, drainage time, hospital stay length were (3.42 ± 1.54) points, (8.52 ± 1.20) h, (7.42 ± 2.43) d, (6.25 ± 2.13) h, (7.58 ± 2.31) d in the study group, and (5.76 ± 1.43) points, (12.42 ± 1.42) h, (10.12 ± 2.52) d, (10.08 ± 2.42) h, (10.16 ± 3.44) d in the control group, there were significant differences between the two groups ( t values were 4.31-14.53, all P<0.05). The incidence of complications was 6.25% (3/48) in the study group, and 16.67% (8/48) in the control group, there was significant difference between the two groups ( χ2=6.56, P<0.05). The total score of FACT-B, joint activity at 1 month after operation, effective cough and turning times with 1 d after operation were (84.48 ± 11.75) points, (28.75 ± 2.58)°, (6.64 ± 2.35) times, (6.10 ± 1.45) times in the study group, and (75.26 ± 11.88) points, (15.38 ± 4.76)°, (3.43 ± 1.25) times, (4.35 ± 1.13) times in the control group, there were significant differences between the two groups ( t values were 3.82-17.11, all P<0.05). There was significant difference in the muscle strength of grade between the two groups ( Z=1.21, P<0.05). Conclusions:FAS in the early pain management of patients after radical mastectomy can shorten the first activity time of out-of-bed and hospital stay length, relieve pain, promote muscle and joint function exercise, reduce the incidence of complications, and improve the quality of life.

OBJECTIVETo investigate the total in situ apoptotic cell number and the apoptotic situation in erythroid cell and megakaryocytes in patients with myelodysplastic syndromes (MDS).

METHODSApoptosis cell number and the apoptotic situation of erythroid cell and megakaryocytes were analysed on cold embedded bone marrow sections from 25 MDS patients by DNA in situ end labelling (ISEL)/alkaline phosphatase anti-alkaline phosphatase (APAAP) double stained techniques. Fourteen cases of iron deficiency anemia (IDA) were taken as control.

RESULTSMean apoptotic cell numbers in MDS and control group were (39.44 +/- 29.34)/mm(2) and (13.43 +/- 8.39)/mm(2) respectively (P < 0.01). RA/RAS subtypes had a higher apoptosis ratio (47.56 +/- 32.86/mm(2)) than that in RAEB/RAEB-t subtypes (21.87 +/- 13.65/mm(2)) (P < 0.05). Double staining showed similar apoptosis percentage in erythroid cell and megakaryocytes in MDS patients comparing with that of controls (P > 0.05). Some apoptotic cells showing erythroid or megakaryocytic morphologic characteristics expressed no cluster differentiation antigen.

CONCLUSIONOverapoptosis existed in MDS, RA/RAS group had a higher apoptosis ratio than RAEB/RAEB-t group. No obvious increased apoptosis in erythroid cell and megakaryocytes was observed in MDS perhaps due to the loss of surface antigens in later stages of apoptotic cells.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Alkaline Phosphatase ; immunology ; Apoptosis ; Bone Marrow Cells ; cytology ; metabolism ; Child ; DNA ; genetics ; Female ; Humans ; Immunoenzyme Techniques ; In Situ Nick-End Labeling ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; pathology