Objective To investigate the effect of TGF-β1/SMAD signaling pathway on K562 cells growth inhibition caused by HMBA. Methods After establishing the in vitro differentiation model with HMBA on K562 cells, the MTT assay was used to detect the proliferation of K562 cells, the cell cycle profile was detected by flow cytometry, and the mRNA expression of TGF-β1, SMAD3, SMAD4 and EVI1 was measured by RT-PCR assay. Results HMBA could inhibit the proliferation and promote the differentiation of K562 cells obviously, which was time and concentration-dependent, and the 72 h corresponding IC50, was about 2 mmol/L. Within 72 h, flow cytometry assay indicated that the ration of G0-G1 phase cells was up-regulated, and the results of RT-PCR showed that relative mRNA expression of TGF-β1, SMAD3 and SMAD4 at mRNA level was increased gradually while that of EVI1 was decreased gradually. Conclusion HMBA can inhibit K562 cells proliferation through TGF-β1/SMAD signaling pathway.

Objective To detect the synergetic effect and mechanism of arsenic trioxide(As2O3)and Trichostatin A(TSA)during inducing apoptosis of HL-60 cells.Methods MTT method was used to test the proliferation of HL-60 cells.Cell cycle and apoptosis were detected by FCM.Semi-quantitative RT-PCR was used to detect the mRNA expression of Bax and Bcl-2 in the cells treated by As2O3 and(or)TSA.Results As2O3 combined with TSA could inhibit proliferation and induce cell cycle arrest at G0 and G1.The percent of apoptosis induced by combination of As2O3 and TSA was obviously higher than that of either As2O3 or TSA.Bax gene expression was increased,while Bcl-2 gene expression was decreased,Bax/Bel-2 ratio was up-regulated.Conclusion Synergetic effect by As2O3 and TSA is remarkable in inducing apoptosis of HL-60 cells.Cell cycte arrest and Bax/Bcl-2 ratio play an important role in apoptosis of HL-60 cells induced by As2O3,TSA or their combination.

Objective To evaluate the positive effects of cisplatin on sensitivity of human glioma U251 to tumor necrosis factor-related apoptosis inducing ligand and to investigate the potential mechanism. Methods The expression of green fluorescent protein (GFP) in U251 which was transfected with pAdxsi-GFP-TRAIL was observed by inverted fluorescent microscope ×400) and to ascertain the MOI. The proliferation inhibition was studied by MTT method. Morphological change was detected through inverted florescent microscope and the Hoechst33342 staining assay was used to verify whether cell apoptosis could be induced or not. The cell apoptosis was also analyzed by flow cvtometry with propidium iodide staining. Semi-quantitative RT-PCR was introduced to detect the mRNA expression of apoptosis related gene.Results The expression of TRAIL mRN A was significantly upregulated after transfection. Compared with treatment group of cisplatin and TRAIL alone, the proliferation of U2S1 was significantly inhibited in the cisplatin sensitizing TRAIL group (P < 0.05 ). Nuclear shrinkage and pyknosis fragmentation were observed by Hoechst 33342 staining assay; Apoptotic peak was detected from the results of flow cvtometry and there were significant differences between the sensitizing group and the other two groups ( P < 0.05 ) ; Moreover, the relatively high expression of TRAIL, DR5, caspaseS and down - regulated survivin genes were also observed. There was no significant changes in DR4 expression. Conclusion Cisplatin could extremely enhance the sensitivity of U251 cells to TRAIL And the potential mechanism may related to the increase of TRAIL, DRS, caspaseS genes while the reduction of surivivin gene.

Objective To investigate the reversal effect of the monomer of traditional Chinese medicine on muhidrug resistance(MDR) and its possible mechanism in K562/ADM cell line in vitro. Methods With different concentrations of baicalin, geniposide administered to K562/ADM cells, the proliferation of K562/ ADM cells was detected by the MTY assay. Expression of mdr-1 mRNA, Topo Ⅱ mRNA was measured by semi-quantitive RT-PCR. Results Thatbaicalin and geniposide could increase the sensitivity of K562/ADM cells to adriamycin, multiples of reversion were 1.95 times and 1.46 times. The proliferation of K562/ADM cells was in-hibited obviously by baicalin and geniposide, the level of mdr-1 mRNA expression was down-regulated and the Topo Ⅱ mRNA was up-regulated(P<0.01 ). Conclusion Baicalin and geniposide may reverse the multi-drag-resistance of K562/ADM cells, which was related to the down-regulation of mdr-1 expression and up-reg-ulation of Topo Ⅱ beta expression.

Objective To explore molecular mechanism of expression of vascular endothelial growth factor (VEGF) mRNA and secretion of VEGF protein in HL-60 cells induced by all-trans refinoic acid (ATRA). Methods MTr method was used to detect the proliferation of HL-60 cells induced by ATRA,cell cycle and CD11b expression in HL-60 cells were detected by flow cytometry. Expression of VEGF, c-myc, by-poxia-inducible factor(HIF)-lα, matrix metalloproteinase (MMP)-9 and MMP-2 mRNA were detected by semi-quantitative RT-PCR. VEGF protein in HL-60 cells culture supernatant was measured by ELISA before and after being induced by ATRA. Results After treatment with ATRA,the proliferation of HL-60 cells was obviously inhibited, CD11b expression increased, trend of granulocyte directional differentiation emerged, and differentiation degree was increasd(P <0. 05) ;expression level of c-my and VEGF mRNA was down-regulated (P < 0. 05), but expression level of HIF-1α mRNA was up-regulated (P < 0. 05). VEGF protein level in HL-60 cells culture supernatant was decreased by blocking the expression of MMP-9 or MMP-2(P <0. 05).Conclusion VEGF expression has positive correlation with c-myc expression,but has negative correlation with HIF-1α expression. MMP-9 and MMP-2 may be the main factors regulating VEGF secretion in HL-60 cells.

Objective To study the molecular mechanism of different sensitivities to apoptosis induced by low concentration of As2O3 in PML-RARα negative HL-60 cells and PML-RARα positive NB4 cells. Meth-ods NB4 and HL-60 cells were cultured with As2O3 for 1 to 4 days; cell proliferation were detected by MTT method; the apoptosis was detected by flow cytometry,Bcl-2,Bax and Fas mRNA were determined by RT-PCR. Results The proliferation of NB4 cells was inhibited obviously by As2O3(1.0 μmol/L)with the induction of apoptosis( P <0.05) ,which was accompanied by the down-regulation of Bcl-2 mRNA expression( P <0.05)and the ratio of Bcl-2/Bax(P <0.05), but there was no obvious variation of Bax and Fas expression( P >0.05). Inhibition of proliferation and apoptosis were not obvious in PML-RARα negative HL-60 cells induced by low concentration As2O3 ( P >0.05), and there was no obvious variation of Bcl-2, Bax, Fas mRNA expres-sion or Bcl/Bax ratio( P >0.05). Conclusion The ratio of Bcl-2/Bax is contributed to the different sensitiv-ities of PML-RARα negative HL-60 cells and positive NB4 cells induced by low concentration of As2O3.

Objective With multi?center investigation, to assess the life quality of patients with maintained hemodialysis (MHD) in Liaoning Province and to explore the relationship among the mineral metabolism, the life quality of the patients with MHD, and the repeated hospitalization within the latest three years. Methods 1192 patients with hemodialysis (at least 3 months) from January to March in 2015 at ten blood purification centers in Liaoning Province were selected for the cross?————————sectional survey. The Kidney Health?related Quality of Life (HRQOL) version 1.3 was used to evaluate the MHD patients' life quality. The total length of hospitalization was divided into four groups: 0 days, 3 to 15 days, 16 to 30 days and above 30 days. Results When serum calcium value ranged from 2.1 to 2.5 mmol/L, kidney?disease component summary (KDCS), mental component summary (MCS), physical component summary (PCS) and SF?36+KDCS corresponded to a higher value (P<0.05). When serum phosphorus value ranged from 1.13 to 1.78 mmol/L, KDCS and SF?36+KDCS corresponded to a higher value (P<0.05). When the calcium phosphorus product value ranged from 40.68 to 49.94, MCS corresponded to a higher value (P<0.05). KDCS showed a linear correlation with age (P<0.001), dialysis age, serum calcium (less than or equal to 2.5 mmol/L) (P<0.05); PCS showed a linear correlation with age (P<0.001) and dialysis age (P<0.05); SF?36+KDCS showed a linear correlation with age (P<0.001), and serum calcium (less than or equal to 2.5 mmol/L) (P<0.05), while age and dialysis age were negatively correlated. The hospitalization days showed a linear correlation with age, dialysis age (P<0.001) and serum phosphorus, calcium phosphorus product value (P<0.05), while dialysis age and calcium phosphorus product value were negatively correlated. Among different groups of total hospitalization days in three years, age, hemodialysis age, serum calcium, serum phosphorus, calcium?phosphorus product value and quality of life values were all statistically significant (P<0.05). Conclusions The life quality of patients with MHD were correlated with serum calcium, phosphorus, calcium and phosphorus product value, iPTH, dialysis age and age, while age and dialysis age were of negative correlation. The total number of hospitalization days in 3 years was closely linearly correlated with age and dialysis age, significantly correlated with serum phosphorus, calcium and phosphorus product value, while dialysis age, calcium and phosphorus product value were in a negative correlation. The total number of hospitalization in 3 years was correlated with the patients' age, dialysis age, serum calcium, serum phosphorus, calcium and phosphorus product value and quality of life.

Objective:To investigate the clinical application effect of enhanced recovery after surgery(ERAS) in the perioperative period of primary repair for children with hypospadias.Methods:Non-randomized controlled study.Retrospective analysis was made on clinical data of 206 children with hypospadias admitted to the Department of Urology, Shanxi Children′s Hospital from March 2018 to March 2022.According to whether the concept of ERAS was adopted for perioperative management of children, they were divided into the ERAS group(109 cases) and the non-ERAS group(97 cases). t-test and Chi-square test were used to analyze the differences between the two groups of patients in hospitalization days, hospitalization costs, pain scores on the first day after surgery, average number of bowel movements within 5 days after surgery, first out-of-bed activity time, hospital satisfaction score, and complication rate. Results:Pediatric patients in the ERAS group showed better outcomes compared to the non-ERAS group: shorter hospital stays[(7.71±1.12) d vs.(21.00±1.07) d], lower hospitalization costs[(10 043.21±1 341.00) yuan vs.(12 993.44±1 081.00) yuan], lower postoperative pain scores on the first day[(5.21±1.32) points vs.(6.09±1.07) points], more frequent bowel movements within 5 days[(5.16±0.65) times vs.(2.91±0.48) times], earlier time to first out-of-bed activity[(5.03±0.61) d vs.(7.12±0.57) d], and higher hospital satisfaction scores[(4.71±0.21) points vs.(4.14±0.15) points], and the differences were statistically significant(all P<0.05).Additionally, the ERAS group had lower rates of fever(6.42%, 7/109) and constipation(3.67%, 4/109) before urinary catheter removal compared to the non-ERAS group[21.64%(21/97) and 28.87%(28/97), respectively](all P<0.05).There were no significant differences in the incidences of complications such as urethral prolapse, infection, delayed wound healing, and bleeding, as well as urethral stricture, urethral fistula, and urethral diverticulum after urinary catheter removal between the two groups(all P>0.05). Conclusions:ERAS protocol applied in the perioperative period of primary repair for children with hypospadias can shorten the hospitalization time, lower the hospitalization costs, and accelerate the rehabilitation of children.It is worth further promotion.

OBJECTIVETo detect the expression of cytokines by acute promyelocytic leukemia (APL) cells before and after exposure to arsenic trioxide.

METHODSDiagnoses were performed according to the FAB cytological classification criteria and cytogenetic criteria. Bone marrow or blood samples from APL patients were collected in heparinized tubes, then primary APL cells were separated by traditional Ficoll-Hypaque density centrifugation and purified after adherence to plastic surfaces. IL-1(beta), IL-6, IL-8, TNF alpha and G-CSF levels in the leukemia cell culture supernatants were detected by ELISA. At the same time, nitro blue tetrazolium (NBT) reduction test was used to detect the differentiation of APL cells.

RESULTSAfter 96 hours exposure to arsenic trioxide, 10 - 6 mol/L in vitro or 10 mg/d in vivo, APL cells showed a significant increase of IL-1(beta) (P < 0.05) and G-CSF (P < 0.05) production, and a significant decrease of IL-6 (P < 0.05) and IL-8 (P < 0.05). However, there was no obvious variation of TNF alpha when compared with APL cells without exposure to arsenic trioxide. On the other hand, the proliferation ratio of APL cells in vitro was statistically correlated to the IL-1(beta) secretion ratio or G-CSF secretion ratio. The cell number ratio in patients with detectable IL-1(beta) or G-CSF was higher than that without detectable IL-1(beta) or G-CSF.

CONCLUSIONIL-1(beta) and G-CSF secretion may play an important role in the proliferation of APL cells after exposure to arsenic trioxide.

Arsenicals ; pharmacology ; Cells, Cultured ; Cytokines ; secretion ; Granulocyte Colony-Stimulating Factor ; secretion ; Humans ; Interleukin-1 ; secretion ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Leukemia, Promyelocytic, Acute ; metabolism ; Oxides ; pharmacology ; Tumor Necrosis Factor-alpha ; secretion

To explore the effects of platelet factor 4(PF4) on hematopoietic reconstitution and its mechanism in syngenic bone marrow transplantation (BMT). The syngenic BMT mice models were established. 20 and 26 h before irradiation, the mice were injected 20 micrograms/kg PF4 or PBS twice into abdominal cavity, then the donor bone marrow nuclear cells (BMNC) were transplanted. On the 7th day, spleen clone forming units (CFU-S) were counted. On the 7th, 14th and 21st day after BMT, the BMNC and megakaryoryocytes in bone marrow tissue were counted and the percentage of hematopoietic tissue and expression level of heparan sulfate in bone marrow tissue were assessed. In PF4-treated groups, the CFU-S counts on the 7th day were higher than those in BMT groups after BMT. The BMNC and megakaryoryocyte counts and the percentage of hematopoietic tissue and heparan sulfate expression level were higher than those in BMT group on the 7th, 14th and 21st day after BMT (P < 0.01 or P < 0.05). PF4 could accelerate hematopoietic reconstitution of syngenic bone marrow transplantation. The promotion of the heparan sulfate expression in bone marrow may be one of mechanisms of PF4.
Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Female ; Hematopoietic Stem Cells ; cytology ; Heparitin Sulfate ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Platelet Factor 4 ; pharmacology ; Radiation-Protective Agents ; pharmacology ; Random Allocation ; Spleen ; cytology ; Stem Cells ; Whole-Body Irradiation