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.

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.

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

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 To investigate the influence of the eye acupuncture therapy in the expressions of brain derived neurophic factor(BDNF)and tyrosine kinase receptor B(TrkB)in penumbra brain tissue of ischemic area in the rats with acute cerebral ischemia reperfusion inj ury,and to explore the related mechanism of protective effect on brain of eye acupunture.Methods 62 SD rats were randomly divided into blank control group (n=11),sham operation group (n=11)and model copy group (n=40).The rats in copy model group were used to establish models with suture method,and the successful model rats (n=32)were randomly divided into model group(n=16)and eye acupuncture group(n=16).The rats in eye acupuncture group were treated with eye acupuncture intervention;the acupoints were chosen according to human acupoint selection method, the liver, the upper-jiao, the down-jiao district,and renal were selected for acupuncture intervention. After reperfusion for 2 h,the acupuncture was performed once every 8 h,lasted for 10 times;30 min after the last intervention,the rats were sacrificed;the regional ischemia penumbra around the parts of the brain tissue was obtained and the expressions of BDNF and TrkB mRNA and protein in brain tissue of the rats were detected with RT-PCR and Western blotting method. Results Compared with blank control group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in sham operation group had no statistical significance(P>0.05);compared with sham operation group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in model group were significantly up-regulated(P<0.01);compared with model group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in eye acupuncture group were significantly up-regulated(P<0.05 or P<0.01).Conclusion The expressions of BDNF and TrkB are increased after cerebral ischemia and reperfusion injury.Eye acupuncture can protect the brain of MCAO/R rats,which may be related to the up-regulation of the expressions of BDNF and TrkB in the corresponding penumbra.

Objective To acquire NF-?B p65 TAD(transcription activation domain) and construct its eukaryotic expression vector and express it in endothelial cells.Methods Human umbilical vein endothelial cells(HUVECs) were cultured and total RNA was extracted.The p65 TAD gene was amplified by RT-PCR.After sequencing,the p65 TAD gene was inserted into the eukaryotic expression vector pEGFP-N1 with the green fluorescence protein,named pEGFP-N1-p65 TAD.pEGFP-N1-p65 TAD was transfected into HUVECs and its expression was observed under fluorescence microscope and analyzed by Western blotting.Results p65 TAD(288-548) was cloned successfully.The constructed plasmid including p65 TAD gene was identical to the designed.p65 TAD gene was expressed successfully in HUVECs.Conclusion The construction of eukaryotic expression vector including p65 TAD gene and its expression in HUVECs are very successful.

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

Objective To investigate the association of gene polymorphism of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) with ulcerative colitis (UC) in Chinese patients. Methods The A+49G transition polymorphism at position 49 (exon 1) and C-318T transition polymorphism at position -318 in promoter of the CTLA-4 gene were determined by polymerase chain reaction with sequence specific primers (PCR-SSP) method in 82 Chinese patients with UC and 204 healthy controls of Han nationality. Results No significant differences in the distribution of genotype and allele frequencies were observed between CTLA-4 C-318T and A+49G gene polymorphisms in UC patients and normal controls. How- ever , comparing with healthy subjects,the frequency of haplotype 2,3 (C -318 -G 49 /T -318 -A 49 ) in UC patients was significantly reduced (26% vs 41%, P