Targeting metabolic vulnerability in mitochondria conquers MEK inhibitor resistance in KRAS-mutant lung cancer

MEK is a canonical effector of mutant KRAS; however, MEK inhibitors fail to yield satisfactory clinical outcomes in KRAS-mutant cancers. Here, we identified mitochondrial oxidative phosphorylation (OXPHOS) induction as a profound metabolic alteration to confer KRAS-mutant non-small cell lung cancer (NSCLC) resistance to the clinical MEK inhibitor trametinib. Metabolic flux analysis demonstrated that pyruvate metabolism and fatty acid oxidation were markedly enhanced and coordinately powered the OXPHOS system in resistant cells after trametinib treatment, satisfying their energy demand and protecting them from apoptosis. As molecular events in this process, the pyruvate dehydrogenase complex (PDHc) and carnitine palmitoyl transferase IA (CPTIA), two rate-limiting enzymes that control the metabolic flux of pyruvate and palmitic acid to mitochondrial respiration were activated through phosphorylation and transcriptional regulation. Importantly, the co-administration of trametinib and IACS-010759, a clinical mitochondrial complex I inhibitor that blocks OXPHOS, significantly impeded tumor growth and prolonged mouse survival. Overall, our findings reveal that MEK inhibitor therapy creates a metabolic vulnerability in the mitochondria and further develop an effective combinatorial strategy to circumvent MEK inhibitors resistance in KRAS-driven NSCLC.

Correlation analysis of heart rate, blood pressure and autonomic nerve function recovery in obstructive sleep apnea hypopnea syndrome patients after uvulopalatopharyngoplasty / 中国医师进修杂志

Objective:To investigate the relationship between heart rate, blood pressure and autonomic nerve function in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) after uvulopalatopharyngoplasty (UPPP).Methods:One hundred patients with OSAHS who underwent UPPP in Wuhan Central Hospital Tongji Medical College, Huazhong University of Science and Technology from July 2018 to July 2019 were selected. According to the disease grade of apnea hypopnea index (AHI), 38 cases were divided into mild group (AHI 5 to 15 times/h) and 62 cases were divided into severe group (AHI>16 times/h). The preoperative and postoperative polysomnography and 24 h dynamic electrocardiogram records was used to monitor the patient′s data, and the quality of sleep was compared before and after treatment in patients with sleep apnea, including: apnea and AHI, the longest apnea time (LAT), the lowest oxygen saturation (L SaO 2) and ratio of time with blood oxygen saturation lower than 90% to total sleep time (TSPO 2 90%); patient′s heart rate, including: maximum heart rate, minimum heart rate and average heart rate; heart rate variability (HRV) and related indexes of patients, including: all sinus RR interval (SDNN), RR interval mean standard deviation (SDANN), root mean square (RMSSD) of adjacent RR interval difference, the percentage of adjacent NN>50 ms in total sinus interval difference (PNN50%) and HRV triangle index; autonomic nerve function of patients, including: high frequency band (HF, 0.05 to 0.15 Hz), low frequency band (LF, 0.01 to 0.05 Hz) and LF/HF; patients′blood pressure, including: systolic and diastolic blood pressure. Results:Compared with those before treatment, AHI, LAT, TSPO 2 90%, SDNN, SDANN, RMSSD, PNN50% and HRV trigonometric index were decreased in mild group and severe group after treatment, L SaO 2 was increased, and there were statistical differences ( P<0.05). Compared with those of mild group, AHI, LAT, TSPO 2 90%, SDNN, SDANN, RMSSD, PNN50% and HRV trigonometric index were increased in severe group before treatment, LSaO 2 was decreased, and there were statistical differences ( P<0.05). In the mild group before treatment, mild group after treatment, severe group before treatment and severe group after treatment, the highest heart rates were (127.22 ± 21.87), (72.26 ± 6.15), (143.71 ± 22.09) and (75.03 ± 8.21) beats/min, the lowest heart rates were (50.18 ± 5.21), (61.27 ± 7.10), (42.18 ± 5.13) and (59.67 ± 6.77) beats/min, and the average heart rates were (71.95 ± 8.36), (62.37 ± 6.28), (85.72 ± 11.04) and (64.30 ± 5.89) times/min. After treatment, the maximum heart rate and average heart rate of mild group and severe group were lower than those before treatment, the lowest heart rate was higher than that before treatment, and there were statistical differences ( P<0.05). In the mild group before treatment, mild group after treatment, severe group before treatment and severe group after treatment, the LF were (1107.61 ± 151.69), (768.42 ± 135.18), (1 307.57 ± 182.30), (770.41 ± 160.25) ms 2, HF were (781.81 ± 91.46), (457.24 ± 72.13), (835.03 ± 152.75), (450.44 ± 94.10) ms 2, LF/HF were 1.76 ± 0.25, 1.35 ± 0.14, 1.98 ± 0.32, 1.38 ± 0.11. After treatment, LF, HF and LF/HF in mild group and severe group were lower than those before treatment ( P<0.05); before treatment, LF, HF and LF/HF rate in severe group were higher than those in mild group ( P<0.05). In the mild group before and after treatment, mild group before and after treatment, the systolic blood pressure were (125.01 ± 15.23), (103.22 ± 17.93), (146.13 ± 21.60), (111.25 ± 23.63) mmHg (1 mmHg = 0.133 kPa), and the diastolic blood pressure were (82.27 ± 11.49), (66.13 ± 10.27), (93.52 ± 16.06), (69.10 ± 14.39) mmHg. After treatment, systolic and diastolic blood pressure in mild group and severe group were lower than those before treatment, and there were statistical differences ( P<0.05); systolic and diastolic blood pressure in severe group were higher than that in mild group before treatment, and there were statistical differences ( P<0.05). LF/HF was positively correlated with AHI, mean heart rate, systolic and diastolic blood pressure ( P<0.05), and negatively with HRV triangle index ( P<0.05). Conclusions:Symptoms of OSAHS patients recover gradually after UPPP, and the recovery of autonomic nerve function is correlated with AHI, heart rate and blood pressure.

Efficacy of weight management combined with uvulopalatopharyngoplasty for obesityrelated obstructive sleep apnea-hypopnea syndrome / 南方医科大学学报

OBJECTIVE@#To evaluate the clinical efficacy of weight management combined with pharyngoplasty for treatment of obesity-related obstructive sleep apnea-hypopnea syndrome (OSAHS).@*METHODS@#Sixty obese patients with OSAHS were randomly assigned into the combined treatment group and control group (@*RESULTS@#After 6 months of treatment, the patients receiving the combined treatment showed significant reductions of BMI, neck circumference and waist circumference as compared with the measurements before treatment and with those in the control group (@*CONCLUSIONS@#Weight management combined with uvulopalatopharyngoplasty can produce a good clinical efficacy for treatment of OSAHS with obesity, and the patients should have strengthened continuous family weight management while receiving surgical treatment.

Effect of tympanic dexamethasone injection in the treatment of different types of sudden deafness / 临床耳鼻咽喉头颈外科杂志

Objective@#The aim of this study is to explore the best administration, timing and efficacy of dexamethasone and Mison in the treatment of different types of sudden deafness. @*Method@#242 cases of sudden deafness first diagnosed in our department were selected. According to the guidelines（2015）, the patients were divided into low frequency descending type （49 cases）, high frequency descending type （66 cases）, flat descending type （71 cases） and total deafness （56 cases）. Different types of patients were randomly divided into tympanic injection group and systemic administration group on the basis of routine treatment. Tympanic injection group was further divided into initial injection group and delayed injection group. Tympanic injection was performed under ear endoscope, once every other day, three times for low frequency descending deafness, and five times for other types of deafness. @*Result@#In comparison of total effective rate, there were significant differences among the three treatments in 49 cases of low frequency descending type, 71 cases of flat descending type and 56 cases of total deafness type （P<0.05）. In 66 cases of high frequency descending type, there was no significant difference among the three treatments （P>0.05）. In the comparison of cure rate, the difference of cure rate among the three treatment methods was also significant in low frequency descending type （P<0.05）. In the other three types of deafness, there was no significant difference among the three treatment methods （P>0.05）. There was no significant difference in the effective rate between men and women （P>0.05） in all patients treated by tympanic injection. There was significant difference in the effective rate of tympanic injection within 7 days of onset and 7 days after onset （P<0.05）. @*Conclusion@#Intratympanic injection of dexamethasone is safe, effective, and easy to use as an initial treatment for low frequency descent, flat, and full deafness, and the sooner the better.

Effect of tympanic dexamethasone injection in the treatment of different types of sudden deafness / 临床耳鼻咽喉头颈外科杂志

The aim of this study is to explore the best administration, timing and efficacy of dexamethasone and Mison in the treatment of different types of sudden deafness. 242 cases of sudden deafness first diagnosed in our department were selected. According to the guidelines（2015）, the patients were divided into low frequency descending type （49 cases）, high frequency descending type （66 cases）, flat descending type （71 cases） and total deafness （56 cases）. Different types of patients were randomly divided into tympanic injection group and systemic administration group on the basis of routine treatment. Tympanic injection group was further divided into initial injection group and delayed injection group. Tympanic injection was performed under ear endoscope, once every other day, three times for low frequency descending deafness, and five times for other types of deafness. In comparison of total effective rate, there were significant differences among the three treatments in 49 cases of low frequency descending type, 71 cases of flat descending type and 56 cases of total deafness type （0.05）. In the comparison of cure rate, the difference of cure rate among the three treatment methods was also significant in low frequency descending type （0.05）. There was no significant difference in the effective rate between men and women （>0.05） in all patients treated by tympanic injection. There was significant difference in the effective rate of tympanic injection within 7 days of onset and 7 days after onset （<0.05）. Intratympanic injection of dexamethasone is safe, effective, and easy to use as an initial treatment for low frequency descent, flat, and full deafness, and the sooner the better.

Blocking Two Signals of Immuno-Activated T Cells from Antigen Presenting Cells Can Prevent Chronic GVHD of Murine / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effects of blocking TCR-CD3 and B7-CD28 signals on immune function of mice with chronic GVHD by using TJU103 and CTLA4-Ig.</p><p><b>METHODS</b>On the basis of foregoing murine model of chronic GVHD, according to interference modes after infusion 6×10 spleen cells of donor mice, the recipients were divided into 5 groups: blank control, cGVHD, TJU103 interference, CTLA4-Ig interference and TJU103+CTLA4-Ig interference groups. The score of clinical manifestation and tissue histopathology were used to evaluate the effects of all the interferences on chronic GVHD.</p><p><b>RESULTS</b>TJU103 and CTLA4-Ig could not influence the formation of the mouse chimera. The analysis of Kaplan survival curve of mice with chronic GVHD showed that the CTLA4-Ig and TJU103+CTLA4-Ig reduced the incidence of chronic GVHD, the TJU103 could delay the occurrence of chronic GVHD, but all the interference factors could not change the severity of chronic GVHD.</p><p><b>CONCLUSION</b>TJU103 can delay the onset time of chronic GVHD, and the CTLA4-Ig can reduce the incidences of cGVHD, the combining use of TJU103 and CTLA4-Ig can significantly reduce the incidence of chronic GVHD, but can not change the severity of chronic GVHD.</p>

Synergistic Killing Effects of Arsenic Trioxide Combined with Itraconazole on KG1a Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of arsenic trioxide combined with itraconazole on proliferation and apoptosis of KG1a cells and its potential mechanism.</p><p><b>METHODS</b>The cell morphology was observed with Wrighe-Giemsa staining; cell survival rate was examined by CCK-8; and colony formation capacity was measured by methylcellulose colony formation test; the flow cytometry was used to analyse the cell apoptosis rate and cell cycle; the protein expressions of BCL-2,caspase-3,BAX,SMO,Gli1 and Gli2 were detected by Western-blot.</p><p><b>RESULTS</b>The arsenic trioxide and itraconazole alone both could inhibit the KG1a cell proliferation in dose-and time-dependent manner. In comparison between single and combined drug-treatment group, both the cell survival rate and the colony number of the single drug-treatment group were significantly lower(P<0.05), and the apoptosis rate was higher in the combined drug-treatment group. In the combined-treatment group, the protein expression of Caspase-3 and BAX was upregulated, while the protein expression of BCL-2,SMO,Gli1 and Gli2 was downregulated.</p><p><b>CONCLUSION</b>Arsenic trioxide combined with itraconazole can inhibit the KG1a cell proliferation and induce apoptosis, which may be related with the inhibition of Hh signaling pathway and upregulation of both Caspase-3 and BAX protein expression, and provided experimental data of arsenic trioxide combined with itraconazole for the treatment of refractory AML.</p>

The clinical analysis of Compound Erlongzuoci pill combined with auricular point stickingand vitamin B12 acupoint injection therapy on nervous tinnitus of middle and old aged / 中国生化药物杂志

Objective To explore the clinical analysis of Compound Erlongzuoci pill combined with auricular point stickingand vitamin B12 acupoint injection therapy on nervous tinnitus of middle and old aged.Methods 80 cases of middle and old aged patients with neurogenic tinnitus treated in our hospital from November 2015 to November 2016 were selected and randomly divided into the control group and the observation group, 40 cases of patients in each group, the control group was given Auricular Pressure Combined with vitamin B12 acupoint injection, the observation group was treated with Compound Erlongzuoci pills combined with auricular point sticking therapy and vitamin B12 acupoint injection,The clinical therapeutic effect of two groups, the tinnitus severity score of patients before and after treatment, the changes of tinnitus loudness and different frequency thresholds were compared between the two groups.Results The total effective rate of the clinical observation group was significantly higher than that of the control group, the difference between the two groups was statistically significant(P<0.05).Before treatment, there is no significant difference between tinnitus severity score, tinnitus loudness and hearing threshold between the two groups,the indexes after treatment in two groups were significantly improved(P<0.05), and the observation group tinnitus severity score, tinnitus loudness and different frequency thresholds were significantly lower than the control group, the difference was statistically significant(P<0.05).Conclusion Compound Erlongzuoci pill combined with auricular senile nervous tinnitus pressure and vitamin B12 acupoint injection therapy can significantly improve the degree of tinnitus patients, the clinical effect is confirmed.

Artificial ossicular chain reconstruction：biocompatibility, properties and laryngecartilage defect repair / 中国组织工程研究

BACKGROUND: Defected Laryngeal cartilage has many alternatives, including autologous cartilage, al ograft cartilage and metal stents. Although these materials can achieve desired outcomes in laryngeal cartilage defect repair, certain limitations exist. OBJECTIVE: To investigate the biocompatibility and properties of artificial ossicular chain reconstruction materials, and to explore the effect of artificial ossicular chain reconstruction materials on laryngeal cartilage defect repair. METHODS: Porous hydroxyapatite otosteon was prepared by high-temperature calcination of hydroxyapatite, fol owed by cultured in bone morphogenetic protein solution extracted from fresh human bone to construct bone morphogenetic protein-hydroxyapatite artificial ossicular chain reconstruction material. And then, the biocompatibility and characteristics of the material were analyzed. Forty adult male New Zealand white rabbits were randomly divided into porous hydroxyapatite group and artificial ossicular chain reconstruction material group (n=20 per group), and underwent repair with porous hydroxyapatite material and bone morphogenetic protein-hydroxyapatite artificial ossicular chain reconstruction material respectively after modeling of laryngeal cartilage defect. RESULTS AND CONCLUSION: There was a significant difference in compressive strength of artificial ossicular chain reconstruction materials with different porosities. No symmetry sphere formed in hol ows of the outer surface of the material, with polygonal appearance and with a pore size of 100-200 μm. There were no obvious adverse reactions in both two groups after implantation, but in the artificial ossicular chain reconstruction material group, numerous fibrous connective tissues and obvious bone nodules appeared, and the degradation rate of the material was faster. These results suggest that the bone morphogenetic protein-hydroxyapatite artificial ossicular chain reconstruction material exhibits good biocompatibility and properties, which wil obtain satisfactory outcomes for laryngeal cartilage defect repair. So, the material holds a great value of clinical application.

Adenosine Al Receptor Mediated Neuroprotection of Shenmai Injection on Rat Cerebral Ischemia/Reperfusion Injury: an Experimental Study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe whether adenosine Al receptor (Al R) mediated neuroprotection of Shenmai Injection (SI) on rat cerebral ischemia/reperfusion (I/R) injury.</p><p><b>METHODS</b>The focal cerebral I/R model was established by middle cerebral artery occlusion (MCAO). Totally 60 successfully modeled rats was divided into 5 groups according to randomized block principle, i.e., the model group, the SI group, the SI + AlR antagonist (1,3-dipropyl-8-cyclopentylxanthine, DPCPX) group, the AlR antagonist control group, and the dimethyl sulfoxide (DMSO) control group, 12 in each group. Besides, a sham-operation group was set up (n =12). SI at 15 mL/kg was peritoneally injected to mice in the SI group immediately after cerebral I/R. Equal volume of normal saline was injected to mice in the model group and the sham-operation group. DPCPX at 1 mg/mL was peritoneally injected to mice in the Al R antagonist control group 30 min before peritoneal injecting SI. DPCPX at 1 mg/kg and DMSO at 1 mL/kg were peritoneally injected to mice in the AlR antagonist control group and the DMSO control group 30 min immediately before cerebral I/R. Rats' neurobehavioral scores were assessed after 24 h reperfusion. The volume of cerebral infarction and Bcl-2 protein expression of cerebral infarction penumbra were also detected. Results Compared with the sham-operation group, neurobehavioral scores, the volume of cerebral infarction, and Bcl-2 protein expression increased (all P <0. 05). Compared with the model group, neurobehavioral scores and the volume of cerebral infarction obviously decreased, but Bcl-2 protein expression increased in the SI group (all P <0. 05). Compared with the SI group, neurobehavioral scores increased, the volume of cerebral infarction was obviously enlarged, and Bcl-2 protein expression was obviously reduced in the A1R antagonist control group (all P <0. 05).</p><p><b>CONCLUSIONS</b>SI's neurobehavioral scores could be partially reversed in the Al R antagonist control group, the volume of cerebral infarction and Bcl-2 protein expression improved. AlR might possibly meditate neuroprotection of SI on MACO mire</p>

Excitotoxic effects of glutamate on cochlear organotypic cultures / 华中科技大学学报（医学）（英德文版）

Glutamate (Glu) is the major afferent excitatory neurotransmitter in the auditory system, and excessive Glu may play an important role in cochlear dysfunction. It is unclear how excessive Glu plays roles in cochlear dysfunction in cochlear organotypic cultures. In this study neonatal rat cochlear organotypic cultures were prepared, and then the cochlear tissues were incubated with a new medium containing specific concentrations of Glu (0.1, 0.5, 1, 10 or 20 mmol/L) for 24 h, or incubated with the medium containing a concentration of 20 mmol/L Glu for 6, 12, 24 or 72 h, respectively. It was found that when the cochlear tissues were cultured for 24 h, the inner hair cells (IHCs) were damaged at the concentration of 0.5 mmol/L Glu, and with the increases of the concentrations, the injury was gradually aggravated, and 20 mmol/L Glu resulted in the significant loss of IHCs. In the 20 mmol/L Glu groups, the stereocilia bundles were missing or disarrayed on a few IHCs after culture for 6 h and the damage effect was time-dependent. The missing of IHCs was more significant in the basal turn of the cochlea than in the middle turn of the cochlea under the same concentration of Glu exposure. These results suggest that excessive exogenous Glu affects the morphology of IHCs, but not affects the outer hair cells (OHCs) in cochlear organotypic cultures, and the excitotoxic effects are different on IHCs of different parts of the cochlea under the same concentration of Glu exposure.

Excitotoxic effects of glutamate on cochlear organotypic cultures / 华中科技大学学报（医学）（英德文版）

Glutamate (Glu) is the major afferent excitatory neurotransmitter in the auditory system, and excessive Glu may play an important role in cochlear dysfunction. It is unclear how excessive Glu plays roles in cochlear dysfunction in cochlear organotypic cultures. In this study neonatal rat cochlear organotypic cultures were prepared, and then the cochlear tissues were incubated with a new medium containing specific concentrations of Glu (0.1, 0.5, 1, 10 or 20 mmol/L) for 24 h, or incubated with the medium containing a concentration of 20 mmol/L Glu for 6, 12, 24 or 72 h, respectively. It was found that when the cochlear tissues were cultured for 24 h, the inner hair cells (IHCs) were damaged at the concentration of 0.5 mmol/L Glu, and with the increases of the concentrations, the injury was gradually aggravated, and 20 mmol/L Glu resulted in the significant loss of IHCs. In the 20 mmol/L Glu groups, the stereocilia bundles were missing or disarrayed on a few IHCs after culture for 6 h and the damage effect was time-dependent. The missing of IHCs was more significant in the basal turn of the cochlea than in the middle turn of the cochlea under the same concentration of Glu exposure. These results suggest that excessive exogenous Glu affects the morphology of IHCs, but not affects the outer hair cells (OHCs) in cochlear organotypic cultures, and the excitotoxic effects are different on IHCs of different parts of the cochlea under the same concentration of Glu exposure.

Preparation and property analysis of a novel dental lithium disilicate glass-ceramic / 实用口腔医学杂志

Objective:To prepare a novel dental lithium disilicate glass-ceramic,and to study its properties.Methods:A2 colored novel glass-ceramic was prepared by sintering technique.The flexural strength,density,elastic modulus,hardness,fracture toughness, translucency and microstructure of the novel glass-ceramic and IPS e.max Press (LT A2,MO1 and HO1 )ceramic ingots were studied. Results:The novel glass-ceramic demonstrated a lower flexural strength (31 5 MPa)than MO ceramic ingot(338 MPa)(P 0.05).The translu-cency parameter of the novel glass-ceramic(21 .2)was higher than that of HO(1 6.5)but lower than that of LT(27.8)and MO(27.5) ceramic ingots(P <0.05).SEMimages showed an interlocking microstructure of rod-shaped Li2 Si2 O5 crystals in all the glass-ceramics investigated.Conclusion:The mechanical properties,translucency and microstructure of the novel glass-ceramic are similar to those of IPS e.max Press ceramic ingots,which can meet the requirements of clinical application.

Honokiol combined with Gemcitabine synergistically inhibits the proliferation of human Burkitt lymphoma cells and induces their apoptosis / 中国实验血液学杂志

This study was aimed to investigate the effect of Honokiol (HNK) combined with Gemcitabine (GEM) on the proliferation and apoptosis of human Burkitt lymphoma Raji cells. Cell proliferation was detected by CCK-8 method to study the role of Honokiol and Gemcitabine in Raji cells. The cell apoptosis and cell cycle status were analyzed by flow cytometry. The level of apoptosis-related protein BCL-2 was measured with Western blot. The results showed that compared with cells treated with mentioned above drugs alone, the proliferative potential of cells in combination group was significantly inhibited (P < 0.01) and the inhibition rate was related to the concentration and action time of HNK; and apoptosis rate markedly increased (P < 0.01), while most Raji cells were arrested at G0/G1 phase and decreased in S phase after treatment with combination of two drugs; the expression of BCL-2 protein decreased (P < 0.01). It is concluded that Honokiol combined Gemcitabine can synergistically inhibit the proliferation, induce cell apoptosis, and down-regulate the expression of BCL-2 in Raji cells. The possible mechanism of synergistic effect may be related with arrest of cell cycle at G0/G1 phase and downregulation of the expression of BCL-2.

The changes of p-Akt/MuRF1/FoxO1 proteins expressions in the conditions of training and immobilization in rats' gastrocnemius muscle / 生理学报

This study was aimed to investigate the changes of muscle protein synthesis and degradation under different movement conditions, so as to provide theoretical basis for muscle atrophy mechanism. Sprague Dawley (SD) rats were randomly divided into control, endurance training (treadmill training), hind limb overhanging and eccentric training (treadmill training, angle -16º) groups. The gastrocnemius muscles of rats were taken and weighed. The muscle was sectioned, and HE staining was employed to determine the cell's cross-sectional area. Protein expression of p-Akt was measured by immunohistochemistry; and the expressions of MuRF1 and FoxO1 were determined by Western blot. The results showed that, compared with control group, hind limb overhanging and eccentric training groups exhibited decreased muscle weight and cross-sectional area, but endurance training group did not show any changes. The expressions of p-Akt in endurance and eccentric training groups, not in hind limb overhanging group, were significantly higher than that in control group. Compared with that of control, MuRF1 protein remained unchanged in endurance training groups, but was increased in eccentric training and hind limb overhanging groups; FoxO1 protein was decreased in endurance training group, but was increased in eccentric training and hind limb overhanging groups. These results indicate that movement (endurance and eccentric training) can activate Akt expression, but does not increase muscle weight, whereas eccentric training and hind limb overhanging can increase the expressions of MuRF1 and FoxO1, and induce amyotrophy, suggesting MuRF1 and FoxO1 are major determinant factors in muscle atrophy.

Synergistic killing effect of arsenic trioxide combined with curcumin on KG1a cells / 中国实验血液学杂志

This study was aimed to explore the effect of arsenic trioxide combined with curcumin on proliferation and apoptosis of KG1a cells and its potential mechanism. The cell survival rate was mesured by MTT; colony formation capacity was examined by methylcellulose colony formation test; flow cytometry was used to analyse the cell surface molecules, cell apoptosis rate and cell cycle; the cell morphology was observed with Wright-Giemsa staining and the protein expression of BCL-2, BAX, PARP was detected by Western blot. The results showed that the phenotype of KG1a cells was CD34(+)CD38(-), while the phenotype of HL-60 cell was CD34(+)CD38(+). The former possessed a stronger colony ability than the latter. Effect of curcumin and arsenic trioxide alone on cell proliferation and inhibition was in dose-dependent manner. Compared with single drug-treatment group, the cell survival rate and colony number were lower, and the apoptosis rate was higher in combined drug-treatment group. Protein expression of BCL-2 and PARP was upregulated, while the protein expression of PARP was downregulated in the combined treatment group. It is concluded that compared with HL-60 cells, KG1a cells are the earlier leukemia stem/progenitor cells. Arsenic trioxide combined with curcumin can effectively inhibit the KG1a cell proliferation and induce apoptosis, which may be associated with the downregulation of BCL-2 and PARP protein expression and the upregulation of BAX protein expression.

Effect of honokiol on proliferation and apoptosis in HL-60 cells and its potential mechanism / 中国实验血液学杂志

This study was aimed to investigate the effect of Honokiol (HNK) on proliferation and apoptosis of acute myeloid leukemia HL-60 cells and its potential mechanism. Inhibitory effect of HNK on the HL-60 cell proliferation was detected by MTT assay. Flow cytometry was used to detect the change of cell cycle and AnnexinV/PI staining was used to detect apoptosis. Western blot was applied to analyze the cell cycle protein (cyclins), cyclin-dependent kinase (CDK), P53, P21, P27, BCL-2, BCL-XL, Bax, caspase-3/9 and proteins for MAPK signal pathway. The results showed that HNK could inhibit the proliferation of HL-60 cells in time- and dose dependent ways. HNK arrested HL-60 cells in G0/G1 phase, and S phase cells decreased significantly (P < 0.05). The expression of cyclin D1, cyclin A, cyclin E and CDK2/4/6 were significantly down-regulated (P < 0.05), the expression of P53 and P21 was significantly upregulated after treating for 24 h with HNK (P < 0.05). After 24 h treatment with HNK, HL-60 cell apoptosis increased significantly with the upregulation of activated caspase-3, -9, BAX expression and the downregulation of BCL-2, BCL-XL expression. The MAPK subfamily, P38 and JNK were not significantly changed, but the expression of MEK1/2-ERK1/2 was significantly downregulated (P < 0.05). It is concluded that HNK arrestes the cells at G0/G1 phase and induces HL-60 cell apoptosis through the intervention of MEK1/2-ERK1/2 signaling pathway.

NVP-BEZ235 inhibits proliferation and colony-forming capability of CD34(+)CD38(-) human acute myeloid leukemia stem cells / 中国实验血液学杂志

This study was aimed to explore the effect of NVP-BEZ235, a dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor, on proliferation, cell cycle and colony forming capability of CD34(+)CD38(-) human acute myeloid leukemia (AML) KG1a cells. Flow cytometry was used to detect expression of CD34 and CD38 on the surface of human AML KG1a cells; Trypan blue assay was used to analyze the effect of NVP-BEZ235 at various concentrations on proliferation of KG1a cells; flow cytometry was performed to examine the cell cycle of KG1a cells after NVP-BEZ235 treatment; Soft agar colony-forming experiment was used to detect the colony forming ability of KG1a cells treated with NVP-BEZ235 at various concentrations. The results indicated that the percentage of CD34(+)CD38(-) AML KG1a cells was (98.02 ± 0.72)%. NVP-BEZ235 (0.125 - 1 µmol/L) inhibited the proliferation of KG1a cells in a time-and dose-dependent manner (P < 0.05) and the 50% inhibition concentrations (IC50) at 24 h and 48 h were 0.597 µmol/L and 0.102 µmol/L, respectively. KG1a cells were arrested at G0/G1 phase after treating with 0.5 µmol/L NVP-BEZ235 for 24 h, it was significantly higher than that of control group (83.2 ± 3.80)% vs (43.47 ± 9.60)% (P < 0.05). KG1a cells treated with NVP-BEZ235 (0 - 1 µmol/L) for 14 d and 21 d, the number of colony decreased respectively from (375.67 ± 21.46) per 2500 KG1a cells and (706.33 ± 87.31) per 2500 KG1a cells to 0, with statistical significance (P < 0.05). It is concluded that NVP-BEZ235 can inhibit proliferation and colony-forming capability of CD34(+)CD38(-) human AML KG1a cells.

Cytotoxicity of Naja Naja Actra Venom Component combined with activated immune cells on leukemia cell line KG1a / 中国实验血液学杂志

This study was aimed to investigate the cytotoxic effect of the Naja Naja Actra Venom Component (NNAVC) combined with activated immune cells on human acute myeloblastic leukemia line KG1a cells. The cytotoxic effects of NNAVC at different concentrations on KG1a cells were measured by CCK-8 method. LDH releasing assay was used to detect the cytotoxic effects of activated immune cells, NNAVC combined with activated immune cells on KG1a cells and the sensitivity of KG1a treated with NNAVC to activated immune cells. The results showed that the inhibitory rate of NNAVC on KG1a cells increased with the concentration enhancement, the cytotoxicity of activated immune cells at the different effector to target (E:T) ratios(6.25:1, 12.5:1, 25:1) on KG1a cells were 12.30%, 24.85% and 52.26%. The cytotoxicity of NNAVC combined with activated immune cells at the different E:T cell ratios (10:1, 20: 1) on KG1a cells were 56.21% and 85.59%, which were higher than that of NNAVC or activated immune cells alone. The cytotoxicity of activated immune cells at the E: T cell ratio of 10:1 on KG1a cells treated with NNAVC at different concentrations were 25.65%, 31.33%, 28.63% and 16.78%, respectively, and that at the E:T cell ratio of 20: 1 were 40.62%, 44.70%, 44.62% and 40.72%. It is concluded that:both of NNAVC and activated immune cells have lethal effect on KG1a cells, and the combination of NNAVC and activated immune cells can strengthen their effect on KG1a.

A case report of myelodysplastic/myeloproliferative disease unclassifiable with karyotype aberration of trisomy 8 and JAK2 mutation / 中国实验血液学杂志

This study aimed to investigate the relationship between clinical features of myelodysplastic/myeloproliferative disease, unclassifiable (MDS/MPD-U), karyotype of chromosome and JAK2 mutation in 1 case. The clinical features, karyotype and JAK2 mutation of the patient with MDS/MPD-U were studied by means of bone marrow biopsy, karyotype analysis and ARMS-PCR technique. The results indicated that the typical micromegakaryocytes and thrombocytosis, karyotype aberration of trisomy 8 as well as JAK2 V617F mutation were found in this patient. It is concluded that the patient was diagnosed as MDS/MPD-U with trisomy 8 and JAK2 V617F mutation. The data of this patient will provide evidence for studying correlation of chromosome karyotype aberration with JAK2 V617F mutation and for evaluating prognosis of MDS/MPD-U.