Objective:To explore the prognostic value of serum free light chain in chronic lymphocytic leukemia patients.Methods:Retrospective cohort study was conducted. One hundred and fifty-six newly diagnosed chronic lymphocytic leukemia(CLL) patients in the first affiliated hospital of Nanjing Medical University from January 2016 to December 2020 were included in the retrospective analysis. Among them, there were 106 males and 50 females, with a median age of 60.7 (53.4, 66.0) years old.Serum sample was collected, serum free light chains were detected, and patients were divided into a treatment group (106 cases) and a follow-up group (50 cases) based on the presence of treatment indications.The threshold of serum free light chain(sFLC) was defined by the reference range of the instruction manual and ROC curve. Three indicators were used, including sFLCR, sFLC(κ+λ) and sFLC(κ-λ). Patients were divided into normal sFLCR group ( n=61)and abnormal group( n=95), as well as sFLC (κ+λ) low value group ( n=88) and high value group ( n=68), and sFLC (κ-λ) low value group ( n=64) and high value group ( n=92).The abnormal group and high value group were enrolled as the experimental group, while the normal group and low value group were enrolled as control group. Chi-square test and Fisher′s exact test were used to compare the clinical data, cytogenetics, and molecular biology characteristics of patients in two groups, Kaplan-Meier method was used to analyze the median treatment-free survival (TFS) of the patients, and Cox regression was used to screen the prognostic factors of the patients. Results:The proportion of Rai stage Ⅰ-Ⅳ ( χ2=8.16, P<0.05 and χ2=7.63, P<0.05 and χ2=5.45, P<0.05), Binet stage B-C( χ2=4.11, P<0.05 and χ2=9.43, P<0.05 and χ2=7.34, P<0.05), β 2-microglobulin>3.5 mg/L( χ2=5.13, P<0.05 and χ2=18.3, P<0.05 and χ2=12, P<0.05), ATM gene mutation rate( χ2=6.21, P<0.05 and χ2=4.88, P<0.05 and χ2=5.19, P<0.05), and immunoglobulin heavy chain variable region (IGHV) mutation free rate ( χ2=18.9, P<0.05 and χ2=24.6, P<0.05 and χ2=10.4, P<0.05)in the experimental group were significantly higher than that in control group 1 ( P<0.05). Multivariate analysis indicated that sFLC(κ+λ) ( HR=1.615,95% CI 1.012-2.576, P=0.044), β 2-microglobulin>3.5 mg/L( HR=2.103,95% CI 1.356-3.262, P=0.001) and TP53 deletion and/or mutation( HR=1.892,95% CI 1.082-3.308, P=0.025) were independent prognostic factors affecting the patients time to first treatment(TFT). Conclusions:Serum free light chains can predict the risk of early treatment and have good prognostic significance in newly diagnosed CLL patients.

To achieve rapid and visual detection of Plasmodium vivax,a detection method based on recombinase polymerase amplification(RPA)technology and lateral flow dipstick(LFD)was established and evaluated.Targeting the conserved sequence of the P.vivax 18S rRNA gene(GenBank:DQ660817.1)as the target sequence,primers and probes were designed with Primer Premier 5,and the P.vivax recombinant plasmid(pUCPv)was constructed as the standard.A sensitive and specific RPA-LFD-based rapid visual detection method for P.vivax nucleic acids was established.The plasmid standard was serially diluted 10-fold to concentrations of 1×103,1×102,1×101,1×10?,and 1×10?1 copies/μL for sensitivity testing.To evaluate specificity,whole blood DNA samples from patients infected with Plasmodium falciparum,Plasmodium malariae,Plasmodium ovale,or Leishmania donovani,as well as healthy participants,were tested by RPA-LFD.Additionally,The assay′s accuracy was evaluated by testing whole blood DNA samples from 24 confirmed P.vivax-infected patients.This study successfully established a sensitive,specific,and rapid visual RPA-LFD method for detecting P.vivax nucleic acids.The assay can complete P.vivax detection within 20 minutes under isothermal conditions at 39 ℃,achieving a sensitivity of 1 copy/μL.There is no significant cross reaction with parasites such as other Plasmodium species and L.donovani,and the specificity is 100%.All 24 DNA samples from confirmed P.vivax patients were detected,showing a 100%detection rate.The developed RPA-LFD assay exhibits excellent sensitivity and specificity,requires only simple heating equipment,and is user-friendly.This rapid visual detection method is particularly suitable for P.vivax screening in low-resource settings.

Objective:To evaluate the cost-effectiveness and impact on mortality of health management services for the elderly aged 65 years and older in national essential public health service project.Methods:Based on the data of county-level medical institutions in Henan Province from 2019 to 2024,the Random Forest Method was used to construct a counterfactual framework to predict the hospitalization expenses under the unmanaged scenario,and then the cost-benefit ratio(BCR)and net income were calculated.Time-dependent Cox proportional hazards model was used to evaluate the effect of health management on all-cause mortality and cardiovascular and cerebrovascular disease mortality in the elderly.Results:A total of 962 955 elderly patients were included,451 119(46.85%)were included in the management group.The average hospitalization cost of the management group was significantly lower than that of the non-management group(P<0.05).Except for 2020-2021,BCRS in 2019 and 2022-2024 were 6.34,2.05,4.45 and 6.60,respectively.The risk of all-cause death was reduced by 76.96%,and the risk of cardiovascular and cerebrovascular death was reduced by 75.57%in the elderly patients included in the management group compared with those not included in the management group.Suggestions:It is necessary to establish a health outcomes-based evaluation system and promote the transformation and upgrading of the service model from single chronic disease management to"integrated health services with multi-disease management".

Objective:To evaluate the cost-effectiveness and impact on mortality of health management services for the elderly aged 65 years and older in national essential public health service project.Methods:Based on the data of county-level medical institutions in Henan Province from 2019 to 2024,the Random Forest Method was used to construct a counterfactual framework to predict the hospitalization expenses under the unmanaged scenario,and then the cost-benefit ratio(BCR)and net income were calculated.Time-dependent Cox proportional hazards model was used to evaluate the effect of health management on all-cause mortality and cardiovascular and cerebrovascular disease mortality in the elderly.Results:A total of 962 955 elderly patients were included,451 119(46.85%)were included in the management group.The average hospitalization cost of the management group was significantly lower than that of the non-management group(P<0.05).Except for 2020-2021,BCRS in 2019 and 2022-2024 were 6.34,2.05,4.45 and 6.60,respectively.The risk of all-cause death was reduced by 76.96%,and the risk of cardiovascular and cerebrovascular death was reduced by 75.57%in the elderly patients included in the management group compared with those not included in the management group.Suggestions:It is necessary to establish a health outcomes-based evaluation system and promote the transformation and upgrading of the service model from single chronic disease management to"integrated health services with multi-disease management".

To achieve rapid and visual detection of Plasmodium vivax,a detection method based on recombinase polymerase amplification(RPA)technology and lateral flow dipstick(LFD)was established and evaluated.Targeting the conserved sequence of the P.vivax 18S rRNA gene(GenBank:DQ660817.1)as the target sequence,primers and probes were designed with Primer Premier 5,and the P.vivax recombinant plasmid(pUCPv)was constructed as the standard.A sensitive and specific RPA-LFD-based rapid visual detection method for P.vivax nucleic acids was established.The plasmid standard was serially diluted 10-fold to concentrations of 1×103,1×102,1×101,1×10?,and 1×10?1 copies/μL for sensitivity testing.To evaluate specificity,whole blood DNA samples from patients infected with Plasmodium falciparum,Plasmodium malariae,Plasmodium ovale,or Leishmania donovani,as well as healthy participants,were tested by RPA-LFD.Additionally,The assay′s accuracy was evaluated by testing whole blood DNA samples from 24 confirmed P.vivax-infected patients.This study successfully established a sensitive,specific,and rapid visual RPA-LFD method for detecting P.vivax nucleic acids.The assay can complete P.vivax detection within 20 minutes under isothermal conditions at 39 ℃,achieving a sensitivity of 1 copy/μL.There is no significant cross reaction with parasites such as other Plasmodium species and L.donovani,and the specificity is 100%.All 24 DNA samples from confirmed P.vivax patients were detected,showing a 100%detection rate.The developed RPA-LFD assay exhibits excellent sensitivity and specificity,requires only simple heating equipment,and is user-friendly.This rapid visual detection method is particularly suitable for P.vivax screening in low-resource settings.

Objective:To explore the prognostic value of serum free light chain in chronic lymphocytic leukemia patients.Methods:Retrospective cohort study was conducted. One hundred and fifty-six newly diagnosed chronic lymphocytic leukemia(CLL) patients in the first affiliated hospital of Nanjing Medical University from January 2016 to December 2020 were included in the retrospective analysis. Among them, there were 106 males and 50 females, with a median age of 60.7 (53.4, 66.0) years old.Serum sample was collected, serum free light chains were detected, and patients were divided into a treatment group (106 cases) and a follow-up group (50 cases) based on the presence of treatment indications.The threshold of serum free light chain(sFLC) was defined by the reference range of the instruction manual and ROC curve. Three indicators were used, including sFLCR, sFLC(κ+λ) and sFLC(κ-λ). Patients were divided into normal sFLCR group ( n=61)and abnormal group( n=95), as well as sFLC (κ+λ) low value group ( n=88) and high value group ( n=68), and sFLC (κ-λ) low value group ( n=64) and high value group ( n=92).The abnormal group and high value group were enrolled as the experimental group, while the normal group and low value group were enrolled as control group. Chi-square test and Fisher′s exact test were used to compare the clinical data, cytogenetics, and molecular biology characteristics of patients in two groups, Kaplan-Meier method was used to analyze the median treatment-free survival (TFS) of the patients, and Cox regression was used to screen the prognostic factors of the patients. Results:The proportion of Rai stage Ⅰ-Ⅳ ( χ2=8.16, P<0.05 and χ2=7.63, P<0.05 and χ2=5.45, P<0.05), Binet stage B-C( χ2=4.11, P<0.05 and χ2=9.43, P<0.05 and χ2=7.34, P<0.05), β 2-microglobulin>3.5 mg/L( χ2=5.13, P<0.05 and χ2=18.3, P<0.05 and χ2=12, P<0.05), ATM gene mutation rate( χ2=6.21, P<0.05 and χ2=4.88, P<0.05 and χ2=5.19, P<0.05), and immunoglobulin heavy chain variable region (IGHV) mutation free rate ( χ2=18.9, P<0.05 and χ2=24.6, P<0.05 and χ2=10.4, P<0.05)in the experimental group were significantly higher than that in control group 1 ( P<0.05). Multivariate analysis indicated that sFLC(κ+λ) ( HR=1.615,95% CI 1.012-2.576, P=0.044), β 2-microglobulin>3.5 mg/L( HR=2.103,95% CI 1.356-3.262, P=0.001) and TP53 deletion and/or mutation( HR=1.892,95% CI 1.082-3.308, P=0.025) were independent prognostic factors affecting the patients time to first treatment(TFT). Conclusions:Serum free light chains can predict the risk of early treatment and have good prognostic significance in newly diagnosed CLL patients.

A new method was developed for simultaneous detection of total 19 kinds of organophosphate esters(OPEs)and their diester metabolites(di-OPEs)in human serum(1.0 mL)and urine(1.5 mL)with low volume of samples.The target compounds were determined using ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)after acetonitrile liquid-liquid extraction combined with purification using an ENVI-18 solid-phase extraction(SPE)column.OPEs and di-OPEs were separated using a Shim-pack GIST C18 column(100 mm×2.1 mm,2 μm)with a Shim-pack GIST-HP(G)C18 guard column.An electrospray ionization source(ESI)was employed in mass spectrometry analysis,with positive/negative ion mode using the multiple reaction monitoring(MRM).All target compounds were separated within 15 min,and exhibited good linear relationships in the concentration range of 2-100 ng/mL,with correlation coefficients(R2)above 0.994.The method detection limits(MDL)in serum ranged from 0.001 to 0.178 ng/mL and the MDL in urine ranged from 0.001 to 0.119 ng/mL.The recoveries of the analytes spiked in serum and urine matrices at two concentration levels were 30.5%-126.8%,with the relative standard deviations(RSDs)ranged from 1%to 23%.In addition,paired serum and urine samples from 11 patients were analyzed.For all samples tested,the internal standards of OPEs exhibited recoveries between 61%and 114%,whereas the internal standards for di-OPEs had recoveries ranging from 43%to 103%.OPEs and di-OPEs exhibited high detection frequencies in 22 serum and urine samples.Triethyl phosphate(TEP),tributyl phosphate(TBP),tris(2-ethylhexyl)phosphate(TEHP),tris(2-butoxyethyl)phosphate(TBEP),tris(1-chloro-2-propyl)phosphate(TCIPP),triphenyl phosphate(TPHP),tri-m-tolyl-phosphate(TMTP)and 2-ethylhexyl diphenyl phosphate(EHDPP)were universally detected in all serum samples.TCIPP was identified at the highest concentrations(median 0.548 ng/mL)in serum samples.In urine samples,the detection frequency for 12 kinds of target compounds reached 100%.Notably,TBP emerged as the predominant OPE in urine,demonstrating a median concentration of 0.506 ng/mL.Regarding di-OPEs,bis(2-chloroethyl)phosphate(BCEP)and bis(2-butoxyethyl)hydrogen phosphate(BBOEP)were the most abundant in urine,with median concentrations of 6.404 and 2.136 ng/mL,respectively.The total concentrations of OPEs and di-OPEs in serum and urine were 1.580-3.843 ng/mL and 5.149-17.537 ng/mL,respectively.These results not only confirmed the effectiveness of the method in detection of OPEs and di-OPEs in biological matrices,but also revealed the widespread presence of OPE compounds in human body and pointed to potential exposure risks.

Objective:To investigate the characteristics of fine lymphocyte typing in patients with myelodysplastic sydrome (MDS) and evaluate the changes of immune status during the course of MDS.Methods:A retrospective study was conducted on 73 patients with MDS, including 47 males and 26 females, aged 66 (56, 73) years old, diagnosed at the First Affiliated Hospital of Nanjing Medical University from February 1, 2022 to January 31, 2024. Besides, 30 healthy individuals from the Health Management Center were selected as the control group. The deep lymphocyte typing was detected by flow cytometry. The differences of lymphocyte subset distribution among the groups were compared; Spearman′s rank correlation coefficient was used to analyze the correlation between lymphocyte subpopulation and blast cells or CD34 +cells. Results:As the risk level of revised International Prognostic Scoring System(IPSS-R)increased, the proportion of helper T(Th) cells ( P=0.047) and naivesubpopulation (CD4+CD45RA+CD62L+) ( P=0.021) in the low-risk group, middle-risk group, and high-risk group increased gradually, while memory subpopulation(CD4+CD45RA-) ( P=0.007) gradually decreased; compared to the group with the low amount of gene mutations, the proportion of B lymphocyte cells (8.76% vs 18.41%, P=0.005) and late activated cytotoxic T lymphocyte (Tc) cells (CD8+HLADR+)(21.21% vs 31.69%, P=0.020) decreased in the high group, and natural killer (NK) cells increased (14.98% vs 8.94%, P=0.032). There was a correlation between the percentage of blast cells and the proportion of Th cells ( P=0.022), naive subpopulation ( P=0.045), memory subpopulation ( P=0.017), and highly activated Tc cells(CD8 +CD38 +HLADR +) ( P=0.013). Conclusions:In patients with MDS, the proportion of Th cells and naive subpopulations is increased, while that of memory subpopulations is decreased, which may indicate an increased risk of IPSS-R; the decreased proportion of B cells, late activated Tc cells, and the increased proportion of NK cells may suggest the accumulation of gene mutations. The differences of lymphocyte subpopulation distribution among MDS patients with different IPSS-R risk level and different amount of gene mutations, indicate that there is immune dysregulation during the course of MDS. Fine lymphocyte typing is helpful to evaluate the changes in immune function in MDS patients.

To clone and express the K26 gene of Leishmania isolated from three types of visceral leishmaniasis epidemic ar-eas in China and evaluate its effect on detecting specific antibodies against visceral leishmaniasis.The K26 fragments from Leishmania isolated KS-6,SC6 and JIASHI-1 was synthesized and cloned into pET32a vector.The recombinant plasmid pET32a-K26 was transformed into Escherichia coli BL21 strains and induced by isopropyl-β-D-thiogalactopyranoside(IPTG).The expressed recombinant protein was purified by the His-tagged affinity column(Ni-NTA).Serum samples of 110 visceral leishmaniasis patients were used for evaluating the sensitivity by ELISA.Serum samples from patients with malaria,schisto-somiasis japonica,cystechinococcosis,toxoplasmosis,paragon-imiasis,clonorchiosis and 40 healthy people were used for eval-uating the specificity.Detection results of ELISA were compared with that of rK39 strip of American InBios company.Comparation among three K26 antigens were given by x2 test.The sensitivity of the recombinant K26 protein of KS-6,SC6 and JIASHI-5 strains of Leishmania and rK39 strip test to detect the sera of patients with visceral leishmaniasis was 90.00％(99/110),92.73％(102/110),90.91％(100/110)and 93.64％(103/110),respectively.There was no cross reactivity with malaria(10),schistosomiasis japonica(10),cystechinococcosis(10),toxoplasmosis(5),paragonimiasis(5)and clonorchiosis(5),and 40 sera from healthy people were also negative.The specificity was 100.00％.There was no statistical difference in the sensitivity of the recombinant K26 protein of KS-6,SC6 and JIASHI-1 strains of Leishmania and rK39 strip test,x2 values are 0.97,0.07 and 0.57 respectively and the P values are 0.33,0.79 and 0.45,respectively.There was no statis-tical difference in the sensitivity of three K26 antigens(x2=0.53,P=0.97).Conclusion The recombinant K26 antigen has po-tential application value in the diagnosis of visceral leishmaniasis.

Objective:To analyze the clinical and genetic characteristics of acute myeloid leukemia,myelodysplasia-related(AML-MR)patients and evaluate their prognostic risk stratification,to guide clinical treatment decisions and improve understanding of the biological characteristics and disease progression.Methods:The study analyzed cellular and molecular genetic information of 307 AML-MR patients,diagnosed based on clinical history,bone marrow morphology,cytogenetics,and molecular genetic abnormalities.The risk stratification followed the 2022 ELN guidelines.Results:57 cases(18.6％)met the AML-MR diagnostic criteria based on morphology and clinical history,110 cases(37.2％)met the AML-MR diagnostic criteria based on cytogenetic results,and 210 cases(74.5％)met the AML-MR diagnostic criteria based on molecular testing results.Among different type of mutations,ASXL1 mutation was the most frequent,followed by SRSF2 and BCOR mutations.Except for 2 cases with incomplete data that could not be classified.263(86.2％)of the 305 patients were classified as poor prognosis,20(6.6％)were classified as good prognosis group,and 22(7.2％)were classified as intermediate prognosis group.Conclusion:Molecular genetic information plays a crucial role in diagnosing AML-MR,highlighting the importance of genetics in diagnosis and prognosis.Most AML-MR patients fall into poor prognosis categories,necessitating early intensive and targeted therapy for better survival outcomes.