Objective To explore the mediating effect of triglyceride-glucose(TyG)index on the risk of proteinuria in patients with type 2 diabetes mellitus(T2DM).Methods 734 patients with T2DM who underwent routine physical examination in Quyang Road Community Health Service Center,Shanghai from March 2023 to May 2023 were enrolled.The results of basic information,biochemical indicators,abdominal ultrasound and other results were collected.All patients were divided into the normal group,microproteinuria group,and massiveproteinuria group,and stratification analyses were underwent according to glycated hemoglobin(HbA1c),body mass index(BMI),TyG index,and presence or absence of non-alcoholic fatty liver disease(NAFLD).Factors affecting proteinuria in T2DM patients were analyzed.Multivariate logistic regression was used to analyze the impact of TyG index and NAFLD on proteinuria in type 2 diabetes population.Regression coefficient sequential test was used to analyze whether TyG mediates NAFLD associated proteinuria.Results There were statistically significant differences in age,BMI,urinary creatinine,HbA1c,TyG index,etc.among the normal group,microproteinuria group,and massiveproteinuria group(all P＜0.05);there was no statistically significant difference in gender among the three groups.Multivariate logistic regression analysis showed that taking the HbA1c＜7％and BMI＜24 kg/m2 group as a reference,the patients with HbA1c≥7％and BMI≥24 kg/m2 had the highest risk of proteinuria(P=0.022),followed by the HbA1c≥7％and BMI＜24 kg/m2 group(P=0.039).Taking the TyG index(7.65-8.69)as a reference,the risk of proteinuria in the(9.45-11.90)group was 3.321 times(P＜0.001).The mediation effect analysis showed that the TyG mediated NAFLD associated proteinuria(P＜0.001),with the mediation effect accounting for 55.70％of the total effect.Conclusion TyG index may be an independent risk factor for proteinuria in patients with T2DM,and the prevalence of proteinuria is high in patients with poor control in HbA1c and excessive BMI,and TyG may partially mediate the risk of proteinuria in patients with T2DM.

Objective:To evaluate the diagnostic effects of Xpert MTB/RIF, Fluorescence PCR melting curve and gene chip technology for rapid screening of rifampicin-resistant tuberculosis.Methods:The clinical data of 150 patients diagnosed with tuberculosis by Bactec MGIT 960 liquid culture drug susceptibility in Zhejiang Chinese Medicine and Western Medicine Integrated Hospital from September 2016 to August 2019 were collected, including Xpert MTB/RIF and gene chip results. The isolated and cultured strains from patients were subjected to fluorescence PCR melting curve detection. Using Bactec MGIT 960 drug susceptibility results as the reference, the diagnostic efficacy of Xpert MTB/RIF, Fluorescence PCR melting curve and gene chip technology for rifampicin resistance were analyzed, and the receiver operating characteristic curve (ROC) was drawn for comparative analysis.Results:Take Bactec MGIT 960 as the gold standard, the sensitivity of Xpert MTB/RIF, Fluorescence PCR melting curve and gene chip technology for rifampicin resistance were 88.89% (16/18), 94.44% (17/18), 88.89% (16/18) respectively; the specificity were 96.21% (127/132), 96.21% (127/132), 95.45% (126/132), respectively. There was no statistically significant difference in the sensitivity and specificity among the three detection methods ( P>0.05). The Kappa values of the three molecular methods for detecting rifampicin resistance were 0.794, 0.827 and 0.770, respectively. The three detection methods have good diagnostic value for rifampicin resistance ( P<0.01), but there is no statistically significant difference between the three methods ( P>0.05). There were 8 cases of inconsistent results between the three methods and Bactec MGIT 960 drug sensitivity. Conclusion:Xpert MTB/RIF, Fluorescence PCR melting curve and gene chip technology have comparable ability to detect rifampicin resistance, all of these have high sensitivity and specificity for detecting rifampicin resistance and are suitable for rapid screening.

Objective To compare the application of PCR-fluorescence probe, Bactec MGIT960 and Xpert MTB／RIF in diagnosis of tuberculosis from non-respiratory specimens.Methods Non-respiratory specimens from 225 patients with suspected tuberculosis admitted in Zhejiang Hospital of Integrated Chinese Medicine and Western Medicine from October 2017 to August 2018 were collected.There were 177 cases of tuberculosis and 48 cases of non-tuberculosis confirmed by clinical diagnosis.All specimens were tested with PCR-fluorescence probe, Xpert MTB／RIF and Bactec MGIT960.The clinical diagnostic results were used as the gold standard, and the receiver operating characteristic curve ( ROC) was drawn to evaluate the diagnostic values of three methods.The consistency of PCR-fluorescence probe method with Xpert MTB／RIF assay was analyzed.Results The sensitivity of PCR-fluorescent probe, Xpert MTB／RIF and Bactec MGIT960 in diagnosis of tuberculosis was 53.67%(95／177), 58.76%(104／177) and 31.07%(55／177), respectively.The sensitivity of PCR-fluorescent probe and Xpert MTB／RIF was higher than that of Bactec MGIT 960 culture ( χ2 ＝17.60 and 27.41, P＜0.01), while there was no significant difference between the PCR-fluorescent probe and the Xpert MTB／RIF (χ2 ＝0.93, P＞0.05).The specificity of three methods were 100.00%(48／48), 100.00%(48／48) and 97.92%(47／48), respectively (F＝1.83, P＞0.05).ROC curve analysis showed that the area under the ROC curve ( AUC) of PCR-fluorescent probe, Xpert MTB／RIF, and Bactec MGIT960 was 0.768, 0.794, and 0.645, respectively.The diagnostic value of PCR-fluorescent probe and Xpert MTB／RIF for tuberculosis was significantly higher than that of Bactec MGIT960 (Z＝5.19 and 6.52, P＜0.01); while Xpert MTB／RIF was superior to PCR-fluorescence probe (Z＝2.8, P＜0.05).In various types of specimens , there was no significant difference in the detection rate of tuberculosis between PCR-fluorescent probe method and Xpert MTB／RIF (χ2 ＝0.73, P＞0.05).The PCR-fluorescent probe and Xpert MTB／RIF had a good consistency (kappa＝0.829).Conclusion Xpert MTB／RIF is superior to PCR-fluorescence probe in the detection of tuberculosis in non-respiratory specimens such as tissues and pus, but the two have good consistency.The PCR-fluorescence probe method is economical and practical , and easy to promote, which has a high clinical application prospects.

Objective To investigate the clinico-pathological features of idiopathic membranous nephropathy (IMN ) and the expression of phospholipase A2 receptor (PLA2R ) in elderly patients. Methods A total of 109 elderly patients with IMN confirmed by renal biopsy at Wuxi People's Hospital from July 2008 to February 2015 were included.Data were retrospectively collected. Results (1)Participating patients with IMN had a mean age of (67.3 ± 5.4)years ,and 67.9% of them had hypertension and 65.1% had nephrotic syndrome.Compared with non-elderly patients ,elderly patients had a higher proportion with hypertension (67.9% vs.25.2%)(P＝0.000) ,higher systolic pressure[(143.1 ± 15.2)mmHg vs. (127.3 ± 13.3)mmHg](P＝ 0.000) ,higher diastolic pressure [(88.4 ± 10.0)mmHg vs. (80.2 ± 8.4)mmHg](P＝ 0.000) ,more severe tubulointerstitial lesions [(3.1±1.9)points vs.(2.0±1.9)points](P＝0.000),and lower eGFR[(70.9±22.9)ml·min－1· 1.73 m －2vs. (90.6 ± 27.1 ) ml·min－1·1.73 m －2] ( P ＝ 0.000 ). (2 ) There were more severe tubulointerstitial lesions[(4.7 ± 1.8)points vs. (2.4 ± 1.7)points ,2.9 ± 1.6 points](P ＝ 0.000 , 0.000)and lower eGFR[(50.4 ± 17.4)ml·min－1·1.73 m －2vs. (80.3 ± 19.7)ml·min－1·1.73 m －2, (72.3 ± 21.4)ml·min－1·1.73 m －2](P＝0.000 ,0.000)in elderly patients of pathological stage Ⅱ, compared with patients of pathological stages Ⅰ and Ⅰ-Ⅱ. (3)The rate of positive PLA2R was 82.4%.Patients with positive PLA2R had higher proteinuria[(4.5 ± 2.3)g vs. (2.9 ± 1.1)g](P＝0.042) ,lower eGFR[(66.8 ± 21.8)ml·min－1·1.73 m －2vs. (97.7 ± 16.0)ml·min－1·1.73 m －2](P＝0.000) ,and more severe tubulointerstitial lesions [(3.1 ± 2.0)points vs. (1.7 ± 1.1)points](P＝0.037)than patients with negative PLA2R. (4)Multiple regression analysis showed that PLA2R positive rate(P＝0.008) ,tubulointerstitial lesion(P＝0.000) ,and level of cholesterol(P＝0.025)were negatively correlated with eGFR (R2＝0.572). Conclusions Compared with non-elderly patients , elderly patients with IMN have poorer prognosis as a result of higher blood pressure and more severe tubulointerstitial lesions.Elderly patients with IMN of advanced pathological stages and positive PLA2R have more severe kidney injury and tubulointerstitial lesions ,resulting in poor prognosis.

Objective: Analysis of the molecular characteristics of eosinophilia. Methods: Targeting sequence to 24 patients with chronic eosinophilic leukemia (CEL) with rearrangement of PDGFRA, PDGFRB, or FGFR1 and 62 patients with hyper-eosinophilic syndrome (HES). Mutation annotation and analysis of amino acid mutation using authoritative databases to speculate on possible pathogenic mutation. Results: Thirty-seven kinds of clonal variant were detected from 17 patients with CEL, no recurrent mutation site and hot spot region were found. No pathogenic mutation was detected in 19 patients with PDGFRA rearrangement, but pathogenic mutations of ASXL1, RUNX1 and NRAS were detected from 2 patients with FGFR1 rearrangement who progressed to acute myeloid leukemia and 1 patient with PDGFRB rearrangement who progressed to T lymphoblastic lymphoma, respectively. One hundred and two kinds of clonal abnormalities were detected in 49 patients with HES. The main hot spot mutation regions included: CEBPA Exon1, TET2 Exon3, ASXL1 Exon12, IDH1 Y208C, and FGFR3 L164V. CRRLF2 P224L and PDGFRB R370C point mutations were detected separately in 2 patients with HES who treated with imatinib monotherapy and achieved hematologic remission. Conclusion The pathogenesis of CEL with PDGFRA, PDGFRB or FGFR1 rearrangement is usually single, and the progression of the disease may involve other driver mutation. A variety of genes with hot mutation regions may be involved in the pathogenesis of HES, and some mutation sites are sensitive to tyrosine kinase inhibitors.

Objective To investigate the features and correlation analysis of clinico-pathological and expression of phospholipase A2 receptor(PLA2R)in idiopathic membranous nephropathy. Methods A number of 244 patients of IMN proved by renal biopsy were recruited in Wuxi People's Hospital from July 2008 to February 2015. Data were restrospectively collected. Results In the 244 IMN patients(mean age 54.07 ± 15.22 years,130 males and 114 females),44.3% had hypertension and 62.7% had nephrotic syndrome. Compared with female patients ,male patients had more severe proteinuria and lower eGFR(P < 0.05). Compared with the group of proteinuria < 4 g/24 h,the groups of proteinuria 4~8 g/24 h and > 8 g/24 h had higher level of cholesterol and blood pressure ,lower eGFR and more severe tubulointerstitial lesions(P < 0.05). Pathological stage Ⅰ and Ⅱaccounted for 98%,and there were more severe tubulointerstitial lesions and lower eGFR in the patients of stage Ⅱthan that in the patients of stageⅠandⅠ~Ⅱ(P<0.05). The positive rate of PLA2R accounted for 84.3%. Lower eGFR and more severe tubulointerstitial lesions were found in PLA2R-positive patients than those in PLA2R-neg-ative patients(P < 0.05). Multiple regression analysis showed that tubulointerstitial lesions(B =-7.253),hyper-tension ratio(B=-10.726)and the level of cholesterol(B=-2.077)had negative correlations with eGFR(P<0.01, R2=0.470). Conclusions IMN patients of male gender,grave proteinuria,high pathological stage and positive PLA2R should be treated more actively , since severe tubulointerstitial lesions and kidney injury were more common in those patients.

Objective: To investigate the clinical manifestation, cytogenetics, gene mutations and prognostic factors of chronic neutrophilic leukemia (CNL) . Methods: 16 CNL cases, according to WHO (2016) -definition, were reviewed retrospectively. Identifications of the CSF3R, ASXL1, SETBP1, CALR and MPL mutations were performed by direct sequencing. JAK2 V617F mutation was detected by AS-PCR. Results: Of the 16 CNL patients, the median age was 64 (43-80) years with a male predominance of 75% (12/16) . The median hemoglobin was 114 (81-154) g/L, with median WBC of 41.20 (26.05-167.70) (10⁹/L and median PLT of 238 (91-394) ×10⁹/L.The median level of marrow fibrosis (MF) was 1 (0-3) degree. There was no other cytogenetic abnormalities except t (1;7) (p32;q11) , +21 and 14ps+ for each. All the 16 CNL patients harbored CSF3R T618I mutation. ASXL1 mutations were identified in 81% (13/16) , while SETBP1 mutations were confirmed in 63% (10/16) . The CALR K385fs*47 mutation was found. There was no mutation in JAK2 V617F or MPL in the above 16 patients. The median overall survival (OS) of patients presented with WBC≥50×10⁹/L at diagnosis (11 months) was significantly shorter than of WBC<50×10⁹/L (39 months, P=0.005) . Conclusion CSF3R T618I mutation was specific for CNL. The median OS of CNL patients was 24 months, and WBC≥50×10⁹/L at diagnosis was an unfavorable prognostic factor.

Objective: To study the characteristics of gene mutations in Chinese myelodysplastic syndromes (MDS) patients. Methods: A total of 511 Chinese patients with MDS performed 112-gene targeted sequencing were retrospectively analyzed. Results: Eighty-three distinct mutant genes were found in 511 patients with MDS. Amongst these, the most frequent mutations was associated with epigenetics (50%) , followed by spliceosome (37%) , signal transduction (34%) , transcription factors (24%) and cell cycle/apoptosis (17%) . 439 subjects (86%) had at least one gene mutation. The mean number of mutations in refractory anemia with unilineage dysplasia (RCUD) was 1.25, refractory anemia with multilineage dysplasia (RCMD) was 1.73, refractory anemia with ring sideroblasts (RARS) was 2.79, refractory anemia with excess blasts-1 (RAEB-1) was 2.22, RAEB-2 was 2.34, MDS with isolated 5q- was 2.67, MDS, unclassified (MDS-U) was 2.00. U2AF1 mutant subjects were more likely to have isolated+8[Q<0.001, OR=4.42 (95% CI 2.23-8.68) ]and less likely to have complex karyotypes[Q=0.005, OR=0.22 (95% CI 0.04-0.72) ]. According to the number of gene mutations, all subjects were categorized into three groups, namely group with 0-1 mutation, with 2 mutations and with three or more mutations. There was a significant difference in overall survival (OS) among three groups (P=0.041) . Conclusion About 90% patients with MDS have at least one gene mutation. Genes associated with epigenetics and spliceosome are most common mutated genes in MDS. The increased numbers of gene mutations accompany with disease evolution and associate with poor prognosis.

Objective: To observe the clinical efficacy and safety of the patients of myelodysplastic syndromes-refractory anemia with excess blasts (MDS-REAB) treated with decitabine alone or based on low dose cytarabine (Ara-C) regimen CAG/HAG [aclarubrci (ACR) /homoharring-tonine (HHT) +cytarabine+granulocyte colony stimulating factor (G-CSF) ]. Methods: Totally 121 patients with MDS-REAB were retrospectively analyzed, including 59 patients treated with decitabine alone (20 mg·m^-2·d^-1 for 5 days) , the rest 62 ones treated with low-dose Ara-C-based regimen CAG/HAG. Overall response rate (ORR) , overall survival (OS) and adverse events of the two groups were analyzed and compared retrospectively. Results: The ORR of decitabine alone or CAG/HAG were 66.2% and 56.4% respectively, with no statistically significant differences (χ²=1.185, P=0.276) . Initial response rate detected by the end of first cycle of CAG/HAG was higher than that of decitabine alone (94.3% vs 69.2%) , there was statistically significant difference in the overall comparison of two groups (χ²=7.612, P=0.009) . The median OS of decitabine alone was 19.5 (95% CI 10.5-28.4) months, the median OS of CAG/HAG was 20.3 (95% CI 10.7-29.9) months, with no statistically significant differences (χ²=0.004, P=0.947) . Grade 3-4 cytopenia and infection were the most prevalent adverses of two group patients. Grade 3-4 cytopenia rate of CAG/HAG was higher than that of decitabine alone (100.0% vs 64.4%, P<0.001) . The infection rate detected at third cycle of CAG/HAG was higher than that of decitabine alone (52.9% vs 15.2%, P=0.008) . Conclusion The efficacy of treating MDS-RAEB with decitabine alone or CAG/HAG was equivalent. CAG/HAG treatment came into effect faster, but decitabine alone treatment was safer.

OBJECTIVETo analyze the clinical, laboratory characteristics and PIG-A gene mutations in patients of myelodysplastic syndromes (MDS) with PNH clones.

METHODS218 MDS patients diagnosed from August 2013 to August 2015 were analyzed. The PIG-A gene mutations were tested in 13 cases of MDS with PNH clones, 17 cases of AA-PNH and 14 cases of PNH selected contemporaneously by PCR and direct sequencing.

RESULTS13 (5.96%) MDS patients were detected with PNH clones (13/218 cases). 9 patients were treated with cyclosporin A (CsA). Patients showed hematological improvement (HI). There were significant differences between MDS-PNH and PNH patients in terms of granulocyte clone size, red cell clone size and LDH levels [19.2% (1.0%-97.7%) vs 60.2% (3.1%-98.0%), P=0.007; 4.3% (0-67.2%) vs 27.9% (2.5%-83.6%), P=0.026; 246 (89-2014) U/L vs 1137 (195-2239) U/L, P=0.049], while the differences were not statistically significant in patients between MDS-PNH and AA-PNH patients [19.2% (1.0%-97.7%) vs 23.2% (1.5%-96.0%), P=0.843; 4.3% (0-67.2%) vs 14.4% (1.1%-62.8%), P=0.079; 246 (89-2014) U/L vs 406 (192-1148) U/L, P=0.107]. PIG-A gene mutations were detected in 7 MDS-PNH patients, of them, six were missense mutations, one were frameshift mutation and four cases with the same mutation of c.356G>A (R119Q). The PIG-A gene mutations were also detected in 9/11 AA-PNH patients and 11/14 PNH patients, both of them had the mutation of c.356G>A (R119Q). The PIG-A gene mutations of MDS-PNH, AA-PNH, PNH patients were all small mutations, the majority of those (59%) were missense mutation and mainly located in exon 2.

CONCLUSIONMDS patients with PNH clones had better response to CsA, smaller PNH clone size. The PIG-A gene mutations of MDS-PNH patients mainly located in exon 2, which could be a mutational hotspot of these patients.

Anemia, Aplastic ; genetics ; Clone Cells ; Erythrocytes ; cytology ; Exons ; Granulocytes ; cytology ; Hemoglobinuria, Paroxysmal ; genetics ; Humans ; Membrane Proteins ; genetics ; Mutation ; Myelodysplastic Syndromes ; genetics ; Polymerase Chain Reaction