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Objective:To investigate the effects of hyperthermia on the biological behavior of human laryngeal cancer Hep-2 cisplatin-resistant (Hep-2/CDDP) cell line and its possible mechanism.Methods:Hep-2/CDDP cell line was induced by high impact combined with increasing concentration method. Cell count method was used to detect the cell proliferation ability of Hep-2 parental cell group (Hep-2 cells without cisplatin-resistance and the cells were cultured with RPMI 1640 cultured medium without cisplatin), Hep-2/CDDP cell group and Hep-2/CDDP+cisplatin group (using RPMI 1640 cultured medium including 4 mg/L cisplatin). Hep-2/CDDP cell group and Hep-2 parental cell group were treated with cultured medium including 0, 0.004, 0.04, 0.4, 4, 40 mg/L cisplatin, respectively. The sensitivity of Hep-2/CDDP cells to cisplatin, vincristine and 5-fluorouracil was determined by using methyl thiazolyl tetrazolium (MTT) method. The half inhibitory concentration ( IC50) and resistance index (RI) were also calculated. Hep-2/CDDP cell group was divided into 4 subgroups: the cells in the control group were cultured for 24 h at 37 ℃; the cells in hyperthermia group were treated at 43 ℃ for 2 h and then re-cultured at 37 ℃ for 22 h; the cells in cisplatin group were cultured at 37 ℃ for 24 h in cultured medium containing 4 mg/L cisplatin. The cells in hyperthermia combined with cisplatin group were cultured in cultured medium containing 4 mg/L cisplatin, treated at 43 ℃ for 2 h and then re-cultured at 37 ℃ for 22 h. The effects of hyperthermia combined with cisplatin on the proliferation and early apoptosis of Hep-2/CDDP cells were detected by using MTT and flow cytometry. The interaction of hyperthermia combined with cisplatin on the proliferation and early apoptosis of HEP-2/CDDP cells was observed by using factorial analysis. Western blotting was used to detect the effect of hyperthermia combined with cisplatin on the expressions of wild-type p53 and PI3K in Hep-2/CDDP cells. Hep-2/CDDP cells were divided into 4 groups: the control group (Hep-2/CDDP cells were cultured for 24 h at 37 ℃); chemotherapy group was treated with 12 mg/L vincristine or 9 mg/L 5-fluorouracil; in the hyperthermia group, Hep-2/CDDP cells were treated at 43℃ for 2 h and then re-cultured at 37 ℃ for 22 h; in hyperthermia combined with chemotherapy group, the cells were cultured in a medium containing 12 mg/L vincristine or 9 mg/L 5-fluorouracil, treated at 43 ℃ for 2 h and then re-cultured at 37 ℃ for 22 h. MTT method was used to detect the effect of hyperthermia combined with vincristine and 5-fluorouracil on the proliferation of Hep-2/CDDP cells. Results:Hep-2/CDDP cell line was successfully established. There were no significant differences in the number of cells in Hep-2/CDDP cell group, Hep-2 parental cell line group and Hep-2/CDDP + cisplatin cell group at different time points (all P > 0.05), and the doubling time was 43.8, 40.6 and 43.5 h, respectively. The IC50 of Hep-2 parental cell line group and Hep-2/CDDP cell group to cisplatin was 4.771 mg/L and 42.749 mg/L, respectively, and the RI was 8.960. Hyperthermia combined with cisplatin could inhibit the proliferation of Hep-2/CDDP cells ( F = 327.91, P < 0.05) and promote the early apoptosis of Hep-2/CDDP cells ( F = 724.63, P < 0.05). Factorial analysis showed that hyperthermia combined with cisplatin had an interaction effect on the proliferation and early apoptosis of Hep-2/CDDP cells ( F = 185.68, 472.51, all P < 0.05). Western blotting showed that the relative expression levels of wild-type p53 protein and PI3K protein in the control group, hyperthermia group, cisplatin group and hyperthermia combined with cisplatin group were significantly different ( F = 547.76, 404.44, all P < 0.01). Hyperthermia combined with vincristine or 5-fluorouracil could inhibit the proliferation of Hep-2/CDDP cells ( F = 33.06, 34.61, all P < 0.05). Factorial analysis showed that hyperthermia combined with vincristine and 5-fluorouracil had no interaction effect on the proliferation of Hep-2/CDDP cells ( F = 0.64,0.60, all P > 0.05). Conclusions:Hyperthermia may reverse the resistance of Hep-2/CDDP cell line to cisplatin by upregulating wild-type p53 expression and inhibiting the PI3K pathway. Hep-2/CDDP cell line has cross-resistance to vincristine and 5-fluorouracil. Hyperthermia can increase the sensitivity of Hep-2/CDDP cell line to vincristine and 5-fluorouracil.
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Objective To evaluate the effect of recombinant human erythropoietin (rhEPO) combined with methylprednisolone sodium succinate (MPSS),compared to MPSS alone,in the treatment of neurological function of patients with acute spinal cord injury (SCI).Methods Twenty-one patients presenting in hospital within less than 8 hours after acute SCI were randomly divided into two groups,the control group (10 cases) and the intervention group (11 cases).The control group was treated by MPSS combined with placebo,while the intervention group received MPSS with rhEPO.Both groups received MPSS 30 mg/kg within the first hour,and if the patient was admitted within 4 hours,MPSS would be applied in the treatment with 5.4 mg/kg per hour in the subsequent 23 hours and till 47 hours if the patient was admitted within 4-8 hours after injury.The intervention group received 500 U/kg rhEPO on admission and another 500 U/kg in the next 24 hours,compared with the control group where placebo was used.The evaluation on neurologic function recovery was made on admission,24 h,72 h,one week,2 months and 6 months later,and statistical analysis was performed.Results The change in ASIA score: in the control group,the increase was seen from admission to 6 months after injury in terms of exercise,algesia and tactile sensation ((31.2±6.6) points vs.(57.8±9.8) points,(41.4±9.5) points vs.(64.3±10.6) points, (39.2±6.8) points vs,(61.5±11.3) points),the increase also took place in the intervention group ((29.5±7.2) points vs.(77.4±10.3) points,(39.7±7.2) points vs.(82.3±12.1) points,(37.4±6.2) points vs.(78.6±12.4) points).As time went on,the increase range in the intervention group became larger,compared with the control group.The difference between the two groups in ASLA score was statistically significant (P0.05).Conclusion The application of MPSS combined with rhEPO within 8 hours after acute spinal injury may be more effective than MPSS with placebo in the neurologic dysfunction recovery.
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Objective To determine the level of peripheral blood CD34 positive (CD34+) cells in patients with acute cerebral infarction (ACI),and to explore its clinical significance.Methods The level of peripheral blood CD34+ cells was determined by flow cytometry within 72 hours of onset of patients with acute cerebral infarction (infarct group,n=45),cerebrovascular risk factors in patients without cerebral infarction (high risk group,n=27) and healthy subjects (control group,n=20).The neural function defect score,infarction lesion volume and carotid artery intima-media thickness (IMT) were determined in patients with infarction group.Results The percentages of peripheral blood CD34 cells in infarction group (0.034 ±0.012)％ and the high risk group of patients (0.047±0.009)％ were lower than that of control group(0.063±0.009)％,and were lower in infarction group than in high-risk groups (all P＜0.05).The percentages of peripheral blood CD34+cells were significantly decreased compared with control group (P＜0.05) in infarction patients with mild [(0.047±0.009)％],moderate [(0.036±0.009)％],severe [(0.022±0.007)％] infarction nervous function defect score.Wherein,the percentages were lower in severe group than in the moderate group,moderate group was lower than in mild group (all P＜0.05).The percentages of peripheral blood CD34 cells in infarction patients with small,moderate,large infaret lesion volume were lower than in control group (P＜0.05),wherein,were lower in large group than in moderate group,lower in moderate group than in treatment group (all P＜0.05).Infarction patients were confirmed with carotid atherosclerosis (CAS) by carotid ultrasound.The extent of lesion were divided into carotid artery intimal thickening group [(0.043±0.010)％],carotid artery plaque group [(0.036±0.010)％],and carotid artery stenosis group [(0.023±0.009)％].The levels of peripheral blood CD34+ cells in three groups of patients were decreased compared with control group.The levels were lower in carotid artery stenosis group than in carotid artery plaque group,lower in carotid artery plaque group than in carotid artery intimal thickening group (all P＜0.05).Conclusions The level of peripheral blood CD34+ cells in acute cerebral ischemia is reduced,it can become a sensitive and early indicator of cerebral ischemia,and its level is related to neurologic impairment,infarction size and the degree of carotid artery atherosclerosis.
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Objective To observe the change of peripheral blood CD34+ cell level in patients with acute cerebral infarction, and explore its relationships with cerebrovascular risk factors,neurological function and carotid artery intima-media thickness (IMT). Methods The 45 patients with acute cerebral infarction (onset within 72 h) (infarction group) and 27 patients with cerebr ovascular risk factors but without cerebral infarction (high-risk group) were chosen for the study. The cerebrovascular disease risk factors including history of alcohol abuse, smoking, coronary heart disease, hypertension, diabetes, abnormal levels of serum triglycerides, total cholesterol,low-density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) were recorded in all subjects. The peripheral blood CD34+ cell levels were measured by flow cytometry.The correlations of peripheral blood CD34+ cell level with cerebrovascular disease risk factors were analyzed. The neurological function and carotid artery IMT were recorded in infarction group, and the correlations of peripheral blood CD34+ cell level with neurological function and carotid artery IMT were analyzed. Results (1) The peripheral blood CD34+ cell level was significantly negatively correlated with coronary heart disease, hypertension, diabetes and LDL-C level (r =- 0. 749,-0. 717, - 0. 688, - 0. 764, all P＜0. 01) ; (2) Multiple linear regression analysis showed that peripheral blood CD34+ cell level was an independent relative factor of acute cerebral infarction (P＜0.05); (3) The peripheral blood CD34+ cell level was lower in infarction group than in high-risk group, and was significantly negatively correlated with neurological deficit score (r=-0. 721, P＜0.01) and carotid artery IMT (r= -0. 695, P＜0. 01). Conclusions Peripheral blood CD34+ cell level could be an independent relative factor of acute cerebral infarction; The peripheral blood CD34+ cell level is significantly negatively correlated with neurological function and carotid artery IMT in patients with acute cerebral infarction; And it can be used as cytological marker which reflect early vascular endothelial function in patients with ischemic stroke.