ABSTRACT
To detect the expression of miR-221/222 in serum and plasma cells in patients with monoclonal gammopathy of undetermined significance(MGUS) and multiple myeloma(MM), and to explore the possibility of miR-221/222 as biomarkers in the diagnosis and prognosis predicting of MGUS and MM.Bone marrow and serum samples from 14 patients with newly diagnosed MGUS, 81 patients with newly diagnosed or relapsed MM and 10 controls were collected from Sir Run Run Shaw Hospital of Zhejiang University and Tongde Hospital of Zhejiang Province during January 2013 and December 2015. The expressions of miR-221/222 in serum and in sorted CD138 positive plasma cells were detected by qRT-PCR, and the relative expression of miR-221/222 (Δct) was compared between the groups. Serum levels of miR-221 before and after treatment were compared in both remission group (=22) and refractory group (=13) in MM patients, and its correlation with serum level of β-MG was assessed using Pearson's correlation analysis.Serum levels of miR-221/222 in MGUS and MM groups were significantly higher than those in control group (all<0.01), while miR-221/222 levels in plasma cells were significantly lower in MGUS and MM groups than those in the control group (<0.05 or<0.01). No significant difference in miR-221/222 levels in serum and plasma cells was observed between MGUS group and MM group (all>0.05). There was no correlation between miR-221/222 levels in serum and plasma cells (=0.024 and -0.127, all>0.05), but miR-221 levels were correlated with miR-222 levels in both serum and plasma cells (=0.534 and 0.552, all<0.01). Receiver operating characteristic (ROC) curves showed that the areas under the curve (AUCs) of serum miR-221/222, plasma cell miR-221/222 in diagnosis of MGUS/MM were 0.968, 0.976, 0.801 and 0.727, respectively. There was no significant difference in serum level of miR-221 among MM patients with different paraprotein isotypes (>0.05), but serum level of miR-221 in patients with relapsed MM was higher than that in patients with newly diagnosed MM (<0.01). Compared with the patients with MGUS or MM stageⅠ and Ⅱ, patients with MM stage Ⅲ were of higher serum levels of miR-221 (<0.01). Serum level of miR-221 decreased after chemotherapy in the remission group (=51.5,<0.01), but such decrease was not observed in the refractory group (=67.5,>0.05). Serum level of β-MG was positively correlated with serum level of miR-221 (=0.524,<0.01).miR-221/222 in serum and plasma cells may be biomarkers for early diagnosis of MGUS, and are helpful for diagnosis and efficacy evaluation of MM.
Subject(s)
Humans , Biomarkers , Blood , Bone Marrow , Chemistry , Disease Progression , MicroRNAs , Blood , Monoclonal Gammopathy of Undetermined Significance , Genetics , Multiple Myeloma , Chemistry , Genetics , Myeloma Proteins , Paraproteinemias , Genetics , Prognosis , RecurrenceABSTRACT
Objective To analyse the distribution characteristics of negative pattern of hepatitis B virus Markers (HBV‐M ) in healthy population in Hangzhou district in 2014 ,and provide strategy for the prevention and control of HBV infection in HBV‐M negative population .Methods The HBV‐M (HBsAg ,HBsAb ,HBeAg ,HBeAb and HBcAb) in blood specimens of health examina‐tion population were tested by using ELISA .For 300 cases preserved HBV‐M negative specimens ,HBsAg and HBsAb were detec‐ted by using chemiluminescence immunoassay and HBV‐DNA was detected by using PROCLEIX ULTRIO? Assay .The viral load of HBV‐DNA reactive sample was quantitatively determined .Results Among 9 143 blood samples ,2 213 samples were HBV‐M negative ,and the negative rate was 24 .20% .The negative rate of male to female was 1∶1 .21 .Using chemiluminescence immunoas‐says and PROCLEIX ULTRIO? Assay simultaneously ,we found one case of low concentration of HBsAg(both HBsAb and HBV DNA nonreactive) ,four cases of low concentration of HBsAb(both HBsAg and HBV DNA nonreactive) ,two cases of HBV‐DNA reactive(HBV‐M negative) .One HBV‐DNA reactive sample could be quantified as 560 IU/mL .Conclusion In HBV‐M (ELISA) negative population of health examination of Hangzhou district ,a few subjects had low concentrations of HBsAg or HBsAb or HBV‐DNA .For HBV‐M negative population ,quantitative detection of HBV‐M and HBV‐DNA before HBV vaccination is recom‐mended to determine w hether they need HBV vaccine and the HBV vaccination plan .
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Objective To identify the hemolysin BL(HBL) gene from Bacillus cereus of patients with endophthalmitis comfirmed by API bacterial identification test strip,and detect its biological activity in vitro.Methods Three pairs of specific primers were designed according to the gene sequence of HBL(B,L1 and L2 components),then the PCR assay were established through condition optimization,and to further detect five Bacillus cereus strains isolated from clinical patients with endophthalmitis.HBL with biological activity was extracted by salt fractionation from a randomly selected strain,and a series of different concentrations of HBL were prepared and acted on sheep red blood cells(SRBC),Vero cells and Hela cells; virulence of HBL was also assessed through observating lethal effect of which on mice with intraperitoneal injection.Results Three genes of HBL were detected in all B.cereus strains from clinical patients; Strong hemolytic activity of HBL showed obvious time-and dose-dependent.In the study,we found the morphological changes of Vero and Hela cells caused by HBL were different,but cell death were the same result with contents released; Within 48 h after infection,lethality of HBL for mice showed 100％ with the concentration of more than 2.0 HU/ml,and was also in a time-and dose-dependent manner.Conclusion HBL isolated from B.Cereus had high hemolysis activity and low concentration.The expression of all BL genes provided a strong basis for the clinical feature of B.Cereus infection,which was developed rapidly and with a poor prognosis.It also provided a new method for rapid diagnosis and molecular epidemiology investigation in clinical.
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Objective To investigate the activity of recombinant Vibrio vulnificus hemolysin (rVvhA) on the apoptosis of J774A.1 cells and the related mechanism. Methods The cytotoxic effect of rVvhA on the growth of J774A.1 cells was identified by MTT, celluar and mitochondrial morphology were observed by transmission electron microscopy, apoptosis or necrosis and mitochondrial membrane potential in J774A.1 cells were measured by flow cytometry, activities of caspase-3 ,-8,-9 were detected by spectrophotometry. Results The viability of J774A.1 cells exposed to rVvhA was inhibited, and it is dependent on dose. Celluar and mitochondrial uhrastructure both occurred to change obviously observed by transmission electron microscopy in J774A.1 treated by 2.0 HU/ml and 3.0 HU/ml rVvhA after 8 hours; and 3.0 HU/ml rVvhA group had a better cytotoxic effect on J774A.1 than that of 3.0 HU/ml rVvhA group. The percentage of apoptosis is (7.80±0.62)%, (12.33±0.12)%, respectively. Besides, the mitochondriai membrane potential also reduced, because the rate of fluorescence which is green increase 1.0% (normal) to 9.8% (2.0 HU/ml rVvhA) and 39.2% (3.0 HU/ml rVvhA). At the same time, the caspase-3, -9 activity increased gradually, but caspase-8 remained unchanging. In J774A.1 cells treated by 3.0 HU/ml rV-vhA + caspase-3 inhibitor(Ac-DEVD-FMK) or caspase-9 inhibitor(Ac-LEHD-FMK), The apoptosis of was reduced to(6.23±3.95)% ,(9.60±3.14)%, and the activity of caspase-3, -9 reduced, too. Conclusion The rVvhA has cytotoxic effect on J774A.1. Mitochondria-mediated apoptosis pathway which is dependent on caspase may be related to apoptosis induced by rVvhA in J774A.1.
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OBJECTIVE To analyze the clinical cases of fungal infections and drug resistance to provide a basis for the treatment of mycotic infection.METHODS A total of 215 cases of fungal strains were identified by API 20C AUX.Drug susceptibility was determined by Rosco slip diffusion.RESULTS In 215 fungal strains of specimens,Candida accounted for 87.9%,of which C.albicans accounted for 37.2%.The yeast-like fungi sensitivity rate to amphotericin B,nystatin and ketoconazole respectively was 100.0%,97.9% and 93.5%.CONCLUSIONS Candida are the most common pathogens in the 215 fungal stains.Yeast-like fungi is sensitive to amphotericin B,nystatin and ketoconazole.