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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 investigate the role of LncRNA ZBED3-AS1 in osteoblast proliferation and differentiation in osteoporotic rats through regulating miR-339-5p/Notch 1.Methods:The rat models of sham operation (Sham) group and model (Model) group were established, and the bone mineral density (BMD) of rats was examined. Rat osteoblasts were isolated and the expression of ZBED3-AS1 and miR-339-5p was detected by qRT-PCR. The osteoblasts of rats in Sham group and Model group were divided into different groups and transfected. CCK8, alizarin red (AR-S) staining and alkaline phosphatase (ALP) staining were used to detect the proliferation and differentiation ability of cells in each group. The distribution of ZBED3-AS1 in cells was determined by FISH assay. Double luciferase report confirmed the relationship between ZBED3-AS1 and miR-339-5p as well as miR-339-5p and Notch 1.Western blot was used to detect the expression of Notch pathway related factors.Results:The bone mineral density of femur in Model group was significantly lower than that in Sham group ( P=0.0057) . Compared with Sham group, the expression of ZBED3-AS1 in osteoblasts of Model group was lower, while that of miR-339-5p was higher (all P<0.05) . Overexpression of ZBED3-AS1 could promote the proliferation and differentiation of osteoblasts, while knockdown of ZBED3-AS1 could inhibit the proliferation and differentiation of osteoblasts (all P<0.05) . ZBED3-AS1 could regulate miR-339-5p as ceRNA (all P<0.05) . Overexpression of miR-339-5p can inhibit the proliferation and differentiation of osteoblasts, which can be partially saved by overexpression of ZBED3-AS1. Notch 1 was confirmed as a target of miR-339-5p, at the same time, interfering with the expression of ZBED3-AS1/miR-339-5p can affect the expression of Notch-1 protein, and the regulation of ZBED3-AS1/miR-339-5p on osteoblasts may be realized through Notch pathway. Conclusion:ZBED3-AS1 can be used as ceRNA to regulate miR-339-5p, and then affect the proliferation and differentiation of osteoblasts in osteoporotic rats.
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Objective:To investigate the predictive value of KAI1 expression in colon cancer tissues for tumor recurrence.Methods:Ninety-two pathological tissue samples were collected from patients undergoing radical operation for colon cancer in Tangshan People's Hospital from August 2010 to November 2011. According to the results of follow-up, the patients were divided into recurrent group (33 cases) and non-recurrent group (59 cases). KAI1 expression in tumor tissues was detected by immunohistochemistry. χ2 test was used to analyze the relationship between KAI1 expression in colon cancer tissues and clinicopathological characteristics of patients with colon cancer. Spearman correlation test was used to analyze the relationship between KAI1 expression in colon cancer tissues and the recurrence time of patients. Cox proportional hazards model was used to analyze the related factors affecting postoperative recurrence of colon cancer. Results:KAI1 expression in tumor tissues in the recurrent group was lower than that in the non-recurrent group [39.39% (13/33) vs. 62.71% (37/59), χ2 = 4.638, P = 0.031]. KAI1 expression was not associated with patients' gender, age and tumor maximum diameter (all P > 0.05), but related to the tumor differentiation and lymphatic metastasis [high and medium differentiation vs. low differentiation: 70.3% (26/37) vs. 43.6% (24/55), χ2 = 6.324, P =0.012; lymph node metastasis vs. non-lymph node metastasis: 43.2% (19/44) vs. 64.6% (31/48), χ2 = 4.238, P = 0.039]. KAI1 expression in tumor tissues was positively correlated with tumor recurrence time ( r = 0.845, P < 0.05). Cox multivariate analysis showed that the low differentiation of the tumor, lymph node metastasis and negative expression of KAI1 in colon cancer tissues were independent risk factors for recurrence of colon cancer after surgery ( HR = 1.736, 95% CI 1.598-5.391, P = 0.019; HR =1.526, 95% CI 1.175-3.029, P = 0.037; HR = 1.799,95% CI 1.756-5.825, P = 0.013). Conclusion:Low KAI1 expression in colon cancer tissues is closely related to colon cancer recurrence, and the detection of KAI1 expression in colon cancer tissues has certain predictive value for tumor recurrence.
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Objective:To study the astragalus polysaccharides ( APS) effect on immune induced by influenza A virus HA2 eu-karyotic expression vector.Methods: The HA2 encoded by the DNA vaccine vector was efficiently expressed in CHO cells, as determined by reverse transcription polymerase chain reaction ( RT-PCR) and fluorescence analysis.60 rats were divided into six groups randomly,which were immunized with normal saline,pEGFP-N1,pHA2/EGFP+different dose of APS by intramuscular injection.The control sera were collected before injection.After injected the 36th day, sera were collected to analyzing IFN-γ, IL-4 and IgG level.Results:IFN-γ,IL-4 and IgG level of pHA2/EGFP+mAPS group was different from that of pEGFP-N1 group or pHA2/EGFP+lAPS group( P<0.05 ).Conclusion: Middle dose of APS could increase immune induced by influenza A virus HA2 eukaryotic expression vector.
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Objective:To study whether high dose astragalus polysaccharides( APS) could affect the expression of pNS1/EGFP that included influenza A virus(IAV) non-structure protein 1(NS1) gene in the tissue.Methods:pNS1/EGFP was constructured with NS1 of IAV.Sixty Kunming mice were divided into three groups randomly.Each group of mice was injected separately with one of the following:pEGFP-N1, pNS1/EGFP and pNS1/EGFP+APS in intraperitoneal injection.The mice were injected by intramuscular injection twice with a 3-week interval between injections.The serum samples and muscle samples were obtained on day 14 and day 28 after the booster injection.Sera IL-4,sera IFN-γ,muscle caspase-3 and muscle NS1 expression were measured in ELISA,Western blot or RT-PCR.Results:There were no significant difference among the different groups in day 14 expect that IFN-γof pNS1/EGFP+APS were lower(P<0.05).IFN-γlevel or IL-4 level of pNS1/EGFP+APS were lower compared with other groups in day 28.caspase-3 of pNS1/EGFP+APS were lower compared with other groups in day 28.Conclusion:APS could increase the expression of pNS1/EGFP by decreasing the inflammation and apoptosis.
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Objective To explore the change of serum thyroid hormone related indicators and the probability of occurrence of thyroid dysfunction abnormality in the patient with type 2 diabetes mellitus(T2DM).Methods 86 patients with T2DM and 61 age-matched and gender-matched individuals with healthy physical examination as controls were selected and detected serum FT3,FT4 and TSH by the electrochemiluminescence method Results The serum FT3,FT4 and TSH in the T2DM group were 5.09 pmol/L, 17.32 pmol/L and 2.81 mIU/L respectively;which in the normal control group were 4.99 pmol/L,17.24 pmol/L and 2.71 mIU/L respectively,the differences between the two groups had no statistical significance(P >0.05).Among 86 cases of T2DM,29 cases had the serum abnormal TSH with the abnormal rate of 33.7%,which in the control group was 14.8% with statistical difference between the two groups(P <0.05).Among T2DM patients,the TSH abnormal rate of in females was 42.1%,which was higher than 17.2% in males.Conclusion The serum thyroid hormone detection is necessary for the T2DM patients,especially female pa-tients,which is conducive to early screening,prevention and treatment.
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Objective To explore the diagnostic value of four tumor markers analyzed with Logistic regression and receiver operator characteristic (ROC) curve in patients with lung cancer.Methods The serum levels of carcinoembryonic antigen (CEA) 、carbohydrate antigen 125 (CA125),cytokeratin 19 fragment (CYFRA21-1) and neuron specific enolase (NSE) were determined by radioimmunoassay in 112 patients with lung cancer and 74 patients with benign pulmonary disease.Four tumor markers were analyzed by Logistic regression and ROC curve.Results The serum levels of CEA,CA125,CYFRA21-1 and NSE in lung cancer patients were [4.53(2.22-11.53)ng/ml,28.97 (11.39-62.10) U/ml,4.05(2.29-8.18)ng/ml,14.11 (11.35-24.12) ng/ml],respectively,which were significantly higher than those in health adults[2.08 (1.45-2.52) ng/ml,12.90 (9.80-19.44) U/ml,1.53 (1.21-2.17) ng/ml,11.38 (9.07-12.80) ng/ml] (all P ＜ 0.01).According to regression equation Y =1/[1 + EXP (4.902-0.394X1-0.627X2-0.165X3)],the area under the ROC curve (AUC),sensitivity,specificity,and accuracy of new variable Y were 0.915 ± 0.020,79.46％,93.24％,and 84.95％,respectively.Conclusions Application of logistic regression and ROC curve increases diagnostic accuracy in lung cancer.
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UNLABELLED@#The patient was hospitalized for sudden hearing impairment for one day.@*PHYSICAL EXAMINATION@#the blood pressure was 150/90 mm Hg, the tympanic membranes in both ears were complete and otopiesis. Audiogram showed total deafness in the right ear and slight sensorineural deafness at speech frequency and 80 db for high tone air conduction and 70 db for bone conduction at high frequency in left ear. Tympanogram showed "A" type in both ears and the ipsilateral and contralateral acoustic reflex in both ears were not induced. BAEP showed that the V wave threshold on the right was not induced and it was 50 dbnHL on the left. CT showed a limited low density area in the clivus. MRI showed a space-occupying lesion behind the basilar clivus and ahead of brain stem. Pathological examination showed CK(+), EMA(+), S-100(+) according to immunohistochemistry, which was in accordance with chondroid chordoma.@*DIAGNOSIS@#chondroid chordoma of clivus.
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Feminino , Humanos , Pessoa de Meia-Idade , Cordoma , Diagnóstico , Perda Auditiva Súbita , Diagnóstico , Neoplasias Cranianas , DiagnósticoRESUMO
Objective To study cutaneous ultrastructural changes and FERMT1 gene mutations in a patient with Kindler syndrome. Methods Clinical data were collected, and tissue samples obtained from the lesions of poikiloderma were observed by using transmission electron microscopy. Fifteen coding exons and their flanking sequences of the FERMT1 gene were amplified by PCR and DNA sequencing was followed.Results Reduplication of lamina densa was seen between the dermal-epidermal junctions of the lesional skin. The patient was found to be homozygous for a novel splice-site mutation (IVS9 + 1G ＞ A) in FERMT1 gene, and his parents were heterozygous for it. The mutation was undetected in fifty normal control individuals.Conclusions Transmission electron microscopy may serve as an ancillary examination for the diagnosis of Kindler syndrome. The IVS9+1G＞A mutation of FERMT1 gene may contribute to the clinical phenotype of Kindler syndrome in this patient.
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A dual-label time-resolved fluoroimmunoassay was established for simultaneously detecting pepsinogen Ⅰ (PGⅠ) and pepsinogen Ⅱ (PG Ⅱ) in human serum. Two capture monoclonal antibodies, 8003# of PGⅠ and 8101# of PGⅡ, were co-coated in 96 microtitration wells. The counterpart tracer monoclonal antibodies, 8016# of PGⅠ and 8102# of PGⅡ, were labeled with Eu3+ and sm3+-chelates, respectively. The samples were assayed by one-step sandwich protocol with the time-resolved fluorometry. The measurement ranges of PGⅠ were 0.2～300.0 ?g/L with the within-run and between-run precision was 5.2% and 8.1%, and that of PGⅡ were 0.05～55.0 ?g/L with the within-run and between-run precision was 7.1% and 11.7%, respectively. The average recovery rates of PGⅠ and PGⅡ were 96.9% and 103.7%, respectively. The results obtained by the dual-label assay agreed well with those by enzyme-linked immunosorbent assays of PGⅠ and PGⅡ, whose correlation ratio were 0.9426 of PGⅠ and 0.9396 of PGⅡ, respectively. The means of 300 healthy volunteers were (157.3 ? 51.0) ?g/L for serum PGⅠ,(10.6 ? 5.9) ?g/L for serum PGⅡ, and (14.8 ? 4.3) for the PGⅠ/PGⅡ ratio. The normal ranges of serum PGⅠ levels for healthy volunteers were 55.3～259.3 ?g/L, those of serum PGⅡ levels were less than 23 ?g/L, the PGⅠ/PGⅡ ratio was more than 6. The proposed dual-label TRFIA for simultaneous detection of PGⅠ and PGⅡ is a simple, sensitive, and rapid method. It could provide serology high-screening of the samples for gastric diseases and would allow investigations into the possible diagnostic value of analysis in various clinical condition.