Objective To investigate the clinical application value of fluorescence PCR in vitro diagnosis(IVD)reagent kits in the HLA-B27 detection by comparing 2 kinds of HLA-B27 IVD reagent kit approved by CFDA.Methods A total of 573 clinical blood samples were collected and detected for HLA-B27 by the approved reagent kits based on the fluorescence PCR technique and the flow cytometry.The samples with inconsistent testing results by the two kits were further confirmed by the PCR sequencing.At the same time,about 5% samples of the positive results detected by the fluorescence PCR method were extracted for conducting the re-testing.Results Among 573 samples,191 samples were HLA-B27 positive and 382 cases were HLA-B27 negative by flow cytome-try;the same samples had 194 cases of HLA-B27 positive and 379 cases of HLA-B27 negative by real-time PCR.With flow cytome-try as reference of the final results,the positive coincidence rate of the two kinds of kit was 96.33%(184/191),the negative coinci-dence rate was 94.76%(362/382),27 samples had inconsistent results from the two kinds of assay(accounting for 4.71% of the to-tal number of samples),the total coincidence rate was 95.29% [(184+362)/573],the Kappa value was 0.896(P =0.02);the chi-square test P =0.021,the two kinds of testing method had the high consistency,but the differences existed in the testing results. The re-testing results by PCR sequencing(including 27 samples with inconsistent results by two kinds of kit)were entirely consist-ent with the fluorescence PCR testing results.Conclusion Compared with the authority method flow cytometry for HLA-B27 tes-ting in clinic,the fluorescence PCR kit may present more accurate judging ability for the HLA-B27 testing on the basis of ensuring the higher consistency of the testing results,is easier compared with the sample preparation and operating procedures,and has the stronger clinical application value and prospects s.

Objective To observe the clinical Resultsof ambroxol combined with glucocorticoids in the treatment of otitis media with effusion.Methods 81 cases fron June 2015 to 2016 October, were randomly, double-blind principle divided into observation group(42cases)and control group(39cases).The control group glucocorticoid therapy observation group were treated with ambroxol combined with glucocorticoid treatment, pure tone was observed before and after treatment were compared listen valve measured air conduction hearing level, hearing recovery and clinical efficacy.ResultsThe observation group were pure tone hearing valve after treatment was measured air conduction hearing level was significantly higher (P<0.05);drum pipe tobacco functioning proportion of patients in the observation group after treatment was 85.71%, significantly higher than 48.72% (P<0.05);The observation group total effective rate was 92.86%, significantly higher than 74.36% (P<0.05).Conclusion Ambroxol combined with glucocorticoids be effective in treating secretory otitis media, otitis media with effusion improve the clinical efficacy, and easy to operate, and is worthy of clinical application.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; blood ; diagnosis ; Phosphopyruvate Hydratase ; blood ; Prognosis

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; metabolism ; pathology ; Polycomb Repressive Complex 1 ; genetics ; metabolism ; Prognosis ; RNA, Messenger ; genetics

Objective To investigate the relationship between the cell cycle blocked by hyperthermia and the expression of rhythm gene Bmall in gastric cancer MKN28 cells so as to provide the academic evidence in hyperthermia therapy for gastric cancer.MethodsThe MKN28 cells were resuscitated and cultured in vitro.In control group MKN28 cells were cultivated at 37 ℃.In experimental groups MKN28 cells were heated at 43 ℃ for different durations.The cell morphology was observed by microscopy.Methylthiazdyl tetrazolium (MTT) assay was adopted to evaluate the inhibitory effect of the cell line.The flow-cytometry was adopted to observe the influence on the cell cycle.The Bmall mRNA expression was investigated by real-time quantitative reverse transcription-polymerase chain reaction ( RT-PCR).ResultsThe remarkable changes of cell morphology were observed by microscopy after exposure to heating.According to the data of MTT assay,37 ℃ heating could not inhibit the proliferation of MKN28.The inhibitive rates of cell growth after 0.5 h,1 h,l.5 h at 43 ℃ was (21.76±5.46)％,(25.30 ±4.36)％ and (27.62 ± 3.78 )％,respectively.Results from flow-cytometry showed that G0/G1 phase cells in lh at 43 ℃ were remarkably less than those in the control group.However G2/M cells were significantly more than those in the control group.The mRNA expression of Bmall was the lowest when heating lh at 43 ℃ as compared to the control group.ConclusionsHyperthermia could induced the cell cycle changes and the expression of Bma11 in gastric cancer MKN28 cells.

Combined Modality Therapy ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Genetic Therapy ; Hepatitis B ; complications ; drug therapy ; Humans ; Liver Cirrhosis ; drug therapy ; etiology ; Medicine, Chinese Traditional ; Phytotherapy

Objective Non steroidal anti inflammatory drugs(NSAIDs)can induce apoptosis in gastric cancer cell and the mechanism is not clear. We aimed to study the mechanism of selective COX 2 inhibitor Nimesulide induced apoptosis of human gastric cancer line SGC 7901 by detecting the expressions of COX 2 at mRNA level, c myc, Bcl 2 and caspase 3 at protein level. Methods Apoptosis was determined by electronic microscopy, Annexin V FITC staining and flow cytometry. The mRNA of COX 2 was detected by RT PCR. The protein expressions of c myc, Bcl 2 and caspase 3 were examined by immunohistochemistry. Results Nimesulide of 50 ?mol/L at 48 and 72 h, and of 100 ?mol/L and 200 ?mol/L at 24, 48 and 72 h induced apoptosis of gastric cancer cells in a dose and time dependent manner.Their apoptotic rates were 7.51%, 9.86% and 11.58%, 12.45%, 16.66% and 12.21%, 15.38%, 20.28% respectively. It increased c myc and caspase 3 expression and decreased Bcl 2 expression and COX 2 mRNA expression. The positive protein expression rates of Bcl 2, c myc and capase 3 were (20.2?7.6)%,(49.2?15.1)% and (34.6?12.9)% respectively with Nimesulide of 200 ?mol/L at 72 h,while the controls being (44.6?12.1)%, (24.7 ?9.5)% and (14.8?6.4)% the three comparative P

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Immunoglobulin D ; Male ; Middle Aged ; Multiple Myeloma ; classification ; pathology ; Retrospective Studies

Objective To evaluate in vitro effects of propranolol and isoproterenol on proliferation of infantile hemangioma endothelial cells(IHECs)as well as expressions of vascular endothelial growth factors(VEGF and VEGF-2) and human basic fibroblast growth factor (bFGF). Methods The second - third passage endothelial cells derived from the proliferative phase of infantile hemangioma were divided into propranolol and isoproterenol groups. The propranolol group was further classified into 5 groups to be treated with propranolol solutions at concentrations of 10, 15, 20 mg/L, EGM-2 medium (blank control group 1), or EGM-2 medium containing 0.16% DMSO (DMSO group), while the isoproterenol group was classified into 4 groups to be treated with isoproterenol solutions at concentrations of 5, 10, 20 mg/L or EGM-2 medium(blank control group 2). Methyl thiazolyl tetrazolium(MTT)assay was performed to evaluate cellular proliferative activity in these propranolol groups at 24, 48, 72 and 96 hours separately, enzyme-linked immunosorbent assay (ELISA)to measure VEGF, VEGF-2 and bFGF lev els in culture supernatants of IHECs at 24 and 48 hours separately. Results The proliferative activity of IHECs showed no significant differences between 10-, 15- and 20-mg/L propranolol groups at either 24 or 48 hours (H = 1.152, 2.643, respectively, both P > 0.05), or between the blank control group 1, DMSO group, and 10- and 15-mg/L propranolol groups at either 72 or 96 hours, but significantly decreased in the 20-mg/L propranolol group compared with the blank control group 1 at 72 and 96 hours (both P < 0.05). The 24-hour treatments with propranolol or isoproterenol at the above concentrations all affected the expressions of VEGF, VEGF-2 and bFGF to different degrees. At 48 hours, there was a significant decrease in VEGF levels in 15- and 20-mg/L propranolol groups, as well as in VEGF-2 and bFGF levels in 10-, 15- and 20-mg/L propranolol groups compared with the blank control group 1 (all P < 0.05), but a significant increase in VEGF levels in 5-, 10- and 20-mg/L isoproterenol groups compared with the blank control group 2 (all P < 0.05), as well as in VEGF-2 and bFGF levels in the 20-mg/L isoproterenol group compared with the blank control group 2, 5- and 10-mg/L isoproterenol groups (all P < 0.05). Conclusions The treatment with propranolol at certain concentrations for certain durations can suppress the growth of, as well as expressions of VEGF, VEGF-2 and bFGF in, endothelial cells derived from the proliferative phase of infantile hemangioma, whereas that with isoproterenol has opposite effects. The therapeutic mechanism of propranolol in infantile hemangioma may be associated with expressions of β-adrenergic receptors and their downstream signal transduction-related cytokines.