Objective To perform the identification at the subspecies-level of Mycobacterium abscessus(M.abscessus)and analyze its characteristics based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS).Methods Bi-otyper software was used to construct the predicted peak spectrum of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense.The predicted peak spectrum was constructed with expected maximum peak value number of 70 and peak frequency of 100％in the ex-perimental group and control group,respectively.A blind test was performed on 31 strains of M.abscessus that were not used to con-struct predictive peak spectra to evaluate the identification efficiency of predictive peak spectra.FlexAnalysis software was used to sum-marize and analyze the list of mass spectral peak value of M.abscessus,and screen the specific peaks in mass spectra of different sub-species of M.abscessus.The principal component analysis(PCA)algorithm was used to perform the cluster analysis for the data from mass spectrometry of M.abscessus,and explore the feasibility of PCA clustering in distinguishing the subspecies of M.abscessus.Results In the experimental group,96.8％(30/31)of the strains were correctly identified,and one strain of M.abscessus subsp.massiliense with rough colony form was mistakenly identified as M.abscessus subsp.abscessus.In control group,77.4％(24/31)of the strains were correctly identified,but 7 strains of M.abscessus subsp.massiliense were incorrectly identified or unable to be identified.The identification efficiency in the experimental group was significantly better than that in the control group with statistical difference(X2=5.167,P=0.026).M.abscessus subsp.abscessus exhibited three specific peaks(m/z 4 001.67,4 386.81 and 4 963.17),and M.abscessus subsp.massiliense also exhibited three specific peaks(m/z 4 950.48,4 381.78 and 5 214.90).In the PCA 3D scatter plot,the data points of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense were relatively dispersed without obvious clus-tering.The PC A dendrograph could be divided into six branches in which only four branches were composed of a single subspecies.The minimum level value of distance between M.abscessus subsp.abscessus and M.abscessus subsp.massiliense was about 0.1.Conclusion The predicted peak spectrum based on MALDI-TOF MS with the expected maximum peak number of 70 could accurately identify M.abscessus at the subspecies level.The specific peak of mass spectrometry method in this study should be feasible to distinguish the subspecies of M.abscessus subsp.abscessus and the subspecies of M.abscessus subsp.Massiliense,but PCA cluster analysis cannot be used as a means to distinguish M.abscessus subsp.abscessus from M.abscessus subsp.massiliense.

Objective:To investigate the levels of serum heat shock protein 27 (HSP27) in patients with multiple myeloma (MM) and its relationship with disease diagnosis, treatment, and prognosis.Method:Retrospective cohort study. We select 85 newly diagnosed MM patients from the Second Affiliated Hospital of Fujian Medical University from December 1, 2021 to June 30, 2023, including 43 males and 42 females, aged 60 (48, 81) years. Their serum HSP27 levels were measured at the time of initial diagnosis, and 100 healthy individuals who underwent physical examinations during the same period were recruited as controls. The correlation between serum HSP27 levels in MM patients and common clinical indicators such as gender, age, hemoglobin (Hb), serum creatinine (Cr), calcium (Ca), lactate dehydrogenase (LDH), β2-microglobulin levels (β2-MG), and bone marrow plasma cell ratio were analyzed; grouping 85 MM patients according to disease types and stages, and the differences in HSP27 levels before and after treatment in different MM classifications, stages, and treatments were compared; using receiver operating characteristic (ROC) curves to obtain the predictive value of initial HSP27 levels for remission in MM patients after standardized treatment, and calculating the optimal critical value. MM patients were divided into a high HSP27 group of 37 cases and a low HSP27 group of 48 cases based on the cutoff value, and Kaplan-Meier was used to compare the survival differences between the two groups.Result:The serum HSP27 levels in newly diagnosed MM patients were higher than those in healthy individuals [(281.33±27.69) pg/ml, (52.11±8.12) pg/ml, t=73.7, P0.001]. There was no correlation between HSP27 levels and patients' MM classification, gender, age, Hb serum Cr, Ca, LDH, and the proportion of bone marrow plasma cells, but positively correlated with β2-MG levels ( r=0.703, P0.05). There are significant differences in HSP27 levels at different stages of MM and before and after treatment; ROC curve results showed that serum HSP27 levels have certain predictive value for whether MM patients can achieve remission after treatment: sensitivity of 75.0%, specificity of 63.8%, and area under the curve of 0.720; the optimal cut-off value is 281.19 pg/ml. The survival time of the low HSP27 level group is longer than that of the high level group. Conclusion:Serum HSP27 is highly expressed in MM, and is correlated with the disease condition, treatment, and prognosis of MM.

Objective To perform the identification at the subspecies-level of Mycobacterium abscessus(M.abscessus)and analyze its characteristics based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS).Methods Bi-otyper software was used to construct the predicted peak spectrum of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense.The predicted peak spectrum was constructed with expected maximum peak value number of 70 and peak frequency of 100％in the ex-perimental group and control group,respectively.A blind test was performed on 31 strains of M.abscessus that were not used to con-struct predictive peak spectra to evaluate the identification efficiency of predictive peak spectra.FlexAnalysis software was used to sum-marize and analyze the list of mass spectral peak value of M.abscessus,and screen the specific peaks in mass spectra of different sub-species of M.abscessus.The principal component analysis(PCA)algorithm was used to perform the cluster analysis for the data from mass spectrometry of M.abscessus,and explore the feasibility of PCA clustering in distinguishing the subspecies of M.abscessus.Results In the experimental group,96.8％(30/31)of the strains were correctly identified,and one strain of M.abscessus subsp.massiliense with rough colony form was mistakenly identified as M.abscessus subsp.abscessus.In control group,77.4％(24/31)of the strains were correctly identified,but 7 strains of M.abscessus subsp.massiliense were incorrectly identified or unable to be identified.The identification efficiency in the experimental group was significantly better than that in the control group with statistical difference(X2=5.167,P=0.026).M.abscessus subsp.abscessus exhibited three specific peaks(m/z 4 001.67,4 386.81 and 4 963.17),and M.abscessus subsp.massiliense also exhibited three specific peaks(m/z 4 950.48,4 381.78 and 5 214.90).In the PCA 3D scatter plot,the data points of M.abscessus subsp.abscessus and M.abscessus subsp.massiliense were relatively dispersed without obvious clus-tering.The PC A dendrograph could be divided into six branches in which only four branches were composed of a single subspecies.The minimum level value of distance between M.abscessus subsp.abscessus and M.abscessus subsp.massiliense was about 0.1.Conclusion The predicted peak spectrum based on MALDI-TOF MS with the expected maximum peak number of 70 could accurately identify M.abscessus at the subspecies level.The specific peak of mass spectrometry method in this study should be feasible to distinguish the subspecies of M.abscessus subsp.abscessus and the subspecies of M.abscessus subsp.Massiliense,but PCA cluster analysis cannot be used as a means to distinguish M.abscessus subsp.abscessus from M.abscessus subsp.massiliense.

Objective:To investigate the levels of serum heat shock protein 27 (HSP27) in patients with multiple myeloma (MM) and its relationship with disease diagnosis, treatment, and prognosis.Method:Retrospective cohort study. We select 85 newly diagnosed MM patients from the Second Affiliated Hospital of Fujian Medical University from December 1, 2021 to June 30, 2023, including 43 males and 42 females, aged 60 (48, 81) years. Their serum HSP27 levels were measured at the time of initial diagnosis, and 100 healthy individuals who underwent physical examinations during the same period were recruited as controls. The correlation between serum HSP27 levels in MM patients and common clinical indicators such as gender, age, hemoglobin (Hb), serum creatinine (Cr), calcium (Ca), lactate dehydrogenase (LDH), β2-microglobulin levels (β2-MG), and bone marrow plasma cell ratio were analyzed; grouping 85 MM patients according to disease types and stages, and the differences in HSP27 levels before and after treatment in different MM classifications, stages, and treatments were compared; using receiver operating characteristic (ROC) curves to obtain the predictive value of initial HSP27 levels for remission in MM patients after standardized treatment, and calculating the optimal critical value. MM patients were divided into a high HSP27 group of 37 cases and a low HSP27 group of 48 cases based on the cutoff value, and Kaplan-Meier was used to compare the survival differences between the two groups.Result:The serum HSP27 levels in newly diagnosed MM patients were higher than those in healthy individuals [(281.33±27.69) pg/ml, (52.11±8.12) pg/ml, t=73.7, P0.001]. There was no correlation between HSP27 levels and patients' MM classification, gender, age, Hb serum Cr, Ca, LDH, and the proportion of bone marrow plasma cells, but positively correlated with β2-MG levels ( r=0.703, P0.05). There are significant differences in HSP27 levels at different stages of MM and before and after treatment; ROC curve results showed that serum HSP27 levels have certain predictive value for whether MM patients can achieve remission after treatment: sensitivity of 75.0%, specificity of 63.8%, and area under the curve of 0.720; the optimal cut-off value is 281.19 pg/ml. The survival time of the low HSP27 level group is longer than that of the high level group. Conclusion:Serum HSP27 is highly expressed in MM, and is correlated with the disease condition, treatment, and prognosis of MM.

Objective To investigate the association between four single nucleotide polymorphisms(SNP)(rs9340 in MAPK1,rs14804 in NRAS,rs712 and rs7973450 in KRAS)in the 3'UTR of ERK1/2 signaling pathway-related genes and non-small cell lung cancer(NSCLC).Methods A total of 478 NSCLC patients and 480 healthy controls were enrolled in this study.Four SNPs were genotyped by using TaqMan assays.The association between the four SNPs and NSCLC was analyzed.Results The distribution frequency difference of the allele of rs9340 was statistically significant between the control group and the non-small cell squamous cell carcinoma(SCC)group(P = 0.009),suggesting that the G allele of rs9340 may be a protective factor for non-small cell lung squamous cell carcinoma(OR = 0.67,95%CI:0.50～0.91).In addition,in the<50 years age group,the distribution frequency difference of the allele of rs9340 was statistically significant between the control group and the NSCLC group(P = 5.07×10-4),indicating that the G allele of rs9340 may be a protective factor for NSCLC(OR = 0.46,95%CI:0.29～0.72).Conclusion The SNP rs9340 in MAPK1 may be associated with the risk of NSCLC.

AIM: To establish the monoclonal antibody against human B lymphocyte stimulator (hBLyS) by DNA immunization and analyse its characterization. METHODS: The 858 bp DNA fragment of hBLyS was cloned into pcDNA3 plasmids. The cloned insert was identified by both sequence analysis and double digestion of the recombinant plasmid with restriction enzymes Xho I and EcoR I . After the splenocytes from BALB/c mice immunized with the recombinant plasmid of pcDNA3/hBLyS were fused with myeloma cells SP2/0,the hybridoma which can produce monoclonal antibodies against hBLyS were obtained. The specificity of anti-BLyS monoclonal antibody from hybridoma was verified by ELISA, Western blot and flow cytometry. RESULTS: The recombinant mammalian cell expression vector of pcDNA3/hBLyS was constructed, the sequence of the insert gene was identified to be the sequence encoding hBLy S antigen. The culture supernatants of hybridoma 9c10 were tested to be the monoclonal antibody with specificity against hBLyS on human peripheral blood CD3 + T cell activated by hIFN-? by ELISA, Western blot and flow cytometry. CONCLUSION: The monoclonal antibodies against hBLyS with high activity and specificity have been established successfully, and will be an useful tool in the studies of relationship between hBLyS and human autoimmunity diseases.