Discrimination of SARS-CoV-2 omicron variant and its lineages by rapid detection of immune-escape mutations in spike protein RBD using asymmetric PCR-based melting curve analysis.

BACKGROUND: The SARS-CoV-2 Omicron strain has multiple immune-escape mutations in the spike protein receptor-binding domain (RBD). Rapid detection of these mutations to identify Omicron and its lineages is essential for guiding public health strategies and patient treatments. We developed a two-tube, four-color assay employing asymmetric polymerase chain reaction (PCR)-based melting curve analysis to detect Omicron mutations and discriminate the BA.1, BA.2, BA.4/5, and BA.2.75 lineages. METHODS: The presented technique involves combinatory analysis of the detection of six fluorescent probes targeting the immune-escape mutations L452R, N460K, E484A, F486V, Q493R, Q498R, and Y505H within one amplicon in the spike RBD and probes targeting the ORF1ab and N genes. After protocol optimization, the analytical performance of the technique was evaluated using plasmid templates. Sensitivity was assessed based on the limit of detection (LOD), and reliability was assessed by calculating the intra- and inter-run precision of melting temperatures (Tms). Specificity was assessed using pseudotyped lentivirus of common human respiratory pathogens and human genomic DNA. The assay was used to analyze 40 SARS-CoV-2-positive clinical samples (including 36 BA.2 and 4 BA.4/5 samples) and pseudotyped lentiviruses of wild-type and BA.1 viral RNA control materials, as well as 20 SARS-CoV-2-negative clinical samples, and its accuracy was evaluated by comparing the results with those of sequencing. RESULTS: All genotypes were sensitively identified using the developed method with a LOD of 39.1 copies per reaction. The intra- and inter-run coefficients of variation for the Tms were ≤ 0.69% and ≤ 0.84%, with standard deviations ≤ 0.38 °C and ≤ 0.41 °C, respectively. Validation of the assay using known SARS-CoV-2-positive samples demonstrated its ability to correctly identify the targeted mutations and preliminarily characterize the Omicron lineages. CONCLUSION: The developed assay can provide accurate, reliable, rapid, simple and low-cost detection of the immune-escape mutations located in the spike RBD to detect the Omicron variant and discriminate its lineages, and its use can be easily generalized in clinical laboratories with a fluorescent PCR platform.

Total serum vitamin B12 (cobalamin) LC-MS/MS assay as an arbiter of clinically discordant immunoassay results.

OBJECTIVES: Measurement of the serum levels of vitamin B12 (VB12) is key for evaluating VB12 deficiency-dependent anemia. Immunoassay, the major method for determining VB12, tends to give false-normal results because of the presence of anti-intrinsic factor (IF-Ab) or other factors such as heterophilic antibodies et al. This study aimed to develop a liquid chromatography tandem mass spectrometry (LC-MS/MS) method that is helpful for distinguish false normal VB12 results measured by the immunoassay. METHODS: Different forms of VB12 were derivatized into CN-B12, which was collected through solid-phase extraction and analyzed via LC-MS/MS. 236 serum samples were measured both by LC-MS/MS and immunoassay, results were compared, and the IF-Ab effect was evaluated. RESULTS: The LC-MS/MS assay afforded a linear slope from 20 to 4,000 pmol/L for CN-B12. OH-VB12, methyl-VB12, and CoA-VB12 showed recovery within 89.3-109.5%. The intra-assay CV of VB12 was 2.6-4.1%, whereas the total CV was 9.3-9.8%. Passing-Bablok regression between LC-MS/MS and immunoassay results showed that the slope was 1.085 and the intercept was -15.691. The Bland-Altman plot showed that the mean difference and difference% were -34.6 pmol/L and 0.3%, respectively. Inter-rater agreement analysis showed that the linear weighted kappa value was 0.885, implying good agreement between the two methods. However, two samples were falsely elevated and one sample was falsely normal in the immunoassay compared with LC-MS/MS. The LC-MS/MS method helped in the distinction of false-normal VB12 results shown by the immunoassay. CONCLUSIONS: The VB12 LC-MS/MS method can be used as an arbiter of clinically discordant immunoassay results.

Role of HLA class I and II alleles in susceptibility to ankylosing spondylitis in Chinese Han.

OBJECTIVE: The objective of the study was to clarify the associations of HLA class I and II alleles with ankylosing spondylitis (AS) among Chinese Han. METHODS: We performed HLA genotyping and Sanger sequencing for 68 HLA-B*27(-), 62 HLA-B*27(+) AS patients, and 70 controls. Case-control analyses and separate analyses of HLA-B*27(-) patients were performed. One-way ANOVA and Kruskal-Wallis multiple comparisons test were used to analyze the effects of HLA-A\B\C\DRB1\DQB1 alleles on clinical characteristics of HLA-B*27(-) and HLA-B*27(+) patients. RESULTS: In the HLA-B*27(+) group, positive associations were seen with A*11:02, B*27:04, B*27:05, C*02:02, C*12:02, and DRB1*04:01 and negative associations were seen with A*33:03, B*07:02, B*57:01, and C*07:02. The age at onset was greater in HLA-B*27(-) patients than in HLA-B*27(+) patients (30.03 ± 15.15 vs. 23.08 ± 7.79 years). In the HLA-B*27(-) group, those with A*01:01, B*13:01, B*13:02, C*01:02, C*04:01, DQB1*02:01, DQB1*06:01, and DRB1*03:01 had an earlier onset than those without these alleles, while patients carrying B*40:02, C*07:02, C*12:02, C*15:02, DQB1*05:02, and DQB1*05:03 had a delayed onset. In the HLA-B*27(-) group, A*32:01(+), C*08:01(+), and DRB1*04:05(-) women were likely to develop AS. In the HLA-B*27(+) group, DQB1*03:02(+) women may be more likely to develop AS. DRB1*12:02 and HLA-B*27 interacted with the distribution of AS-affected sites. In the HLA-B*27(+) group, DRB1*12:02(+) patients were likely to have peripheral joint involvement. CONCLUSION: HLA class I and II alleles other than HLA-B*27 contribute to AS predisposition and characteristics among Chinese Han patients.

Experimental Characteristics of Patients with Newly Diagnosed Multiple Myeloma.

Background: Multiple myeloma (MM), a malignant plasma cell proliferative disease, makes up to 1% of all cancers and somewhat exceeds 10% of all hematological cancers. Since it affects many organs, the signs and symptoms of myeloma vary greatly. This investigation was carried out to identify the clinical and laboratory characteristics of MM. Method: From January 1, 2014, to June 30, 2020, 169 in-patients who received a MM diagnosis for the first time at China-Japan Friendship Hospital in Beijing had their medical information examined. Results: Among 169 newly diagnosed patients, the median age was 60 years (26-84 years). Seven patients were younger than 40 years, and 16.0% (27/169) were 70 years or older. 40.8% (69/169) had IgG M-protein and 27.2% (46/169) had IgA. 84% (142/169) of patients were in the Durie Salmon stage 3. The major sign and symptoms at diagnosis were fatigue (100/169, 59.2%), bone pain (96/169, 56.8%), and weight loss (34/169, 20.1%). Anemia was present initially in 94.0% (159/169), high erythrocyte sedimentation rate in 92.7% (101/109), and thrombocytopenia in 26.6% (45/169). Similarly, hypercalcemia, renal insufficiency, and hypoalbuminemia were observed in 19.3% (31/161), 27.8%, and 75.7% respectively. Immunoparesis was found in 94% (110/117) of IgG, IgA, or IgM patients, and in 87% (33/38) of light chain myeloma patients. A localized band was found in 78.3% (123/157) of patients upon serum protein electrophoresis while monoclonal protein was detected by immunofixation in 91.5% (139/152) of patients. 4.1% (7/169) of the patients had non-secretory myeloma. The prevalence of light chain myeloma was 22.5% (38/169), and these individuals were more likely than other myeloma patients to have renal insufficiency (50% versus 21%, P < .05). In 84.8% of patients, the bone marrow had 10% or more plasma cells. Conclusion: The notable features that can be concluded from this study are the early onset of myeloma in the Chinese population and an advanced disease stage at the time of diagnosis with most of them accompanying anemia, hypoalbuminemia, and immunoparesis.

A novel method for early detection of colorectal cancer based on detection of methylation of two fragments of syndecan-2 (SDC2) in stool DNA.

BACKGROUND: Methylated SDC2 has been proved as a diagnostic marker for human colorectal cancer (CRC), noninvasive stool DNA-based methylation testing also emerges as a novel approach for detecting CRC. The aim of this study was to evaluate the clinical performance of stool DNA-based SDC2 methylation test by a new qPCR detection reagent for early detection of CRC. METHODS: A new qPCR detection reagent contained two differentially methylated regions in SDC2 CpG islands for the detection of CRC was used in this study. Performance of the SDC2 methylation detection reagent was evaluated by analyzing limit of detection, precision, and specificity. The effect of interfering substances on assay performance was also tested. 339 subjects (102 CRC patients, 50 patients with advanced adenomas, 39 patients with non-advanced adenomas, 18 colitis patients and 130 normal individuals) from the China-Japan Friendship Hospital were evaluated. Approximately 2.5 g of stool sample was collected from each participant. Stool DNA was extracted and bisulfite-converted, followed by qPCR assay, which contained two pairs of primers for the methylation detection of two fragments of the SDC2 gene (named SDC2-A and SDC2-B). The diagnostic value of this test in CRC was evaluated by calculating receiver operating characteristic (ROC) curve, and value of the area under the curve (AUC). RESULTS: The test kit was able to detect methylated SDC2 in stool DNA samples with concentrations as low as 90 copies/µL in 100% of replicates. The sensitivity for detecting CRC by methylated SDC2-A alone was 85.29% (95% CI 77.03-91.00%) with a specificity of 96.15% (95% CI 91.08-98.58%). The sensitivity by methylated SDC2-B alone was 83.33% (95% CI 74.82-89.42%) with a specificity of 97.69% (95% CI 93.14-99.51%). However, when methylated SDC2-A and methylated SDC2-B were combined, the sensitivity for CRC detection improved to 87.25% (95% CI 79.27-92.53%) with a specificity of 94.62% (95% CI 89.11-97.56%). Further, the detection reagent achieved ROC-AUC 0.874 (95% CI 0.822-0.927) for SDC2-A, 0.906 (95% CI 0.859-0.952) for SDC2-B, and 0.939 (95% CI 0.902-0.977) for SDC2-Combine A&B. CONCLUSIONS: This study validated the capability of stool DNA-based SDC2 methylation test for early screening of CRC, and combined detection of two fragments of SDC2 gene could improve detection sensitivity.

Rapid detection of the irinotecan-related UGT1A1*28 polymorphism by asymmetric PCR melting curve analysis using one fluorescent probe.

BACKGROUND: Determination of UGT1A1 (TA)n polymorphism prior to irinotecan therapy is necessary to avoid severe adverse drug effects. Thus, accurate and reliable genotyping methods for (TA)n polymorphism are highly desired. Here, we present a new method for polymerase chain reaction (PCR) melting curve analysis using one fluorescent probe to discriminate the UGT1A1*1 [(TA)6 ] and *28 [(TA)7 ] genotypes. METHODS: After protocol optimization, this technique was applied for genotyping of 64 patients (including 23 with UGT1A1*1/*1, 22 with *1/*28, and 19 with *28/*28) recruited between 2016 and 2021 in China-Japan Friendship Hospital. The accuracy of the method was evaluated by comparing the results with those of direct sequencing and fragment analysis. The intra- and inter-run precision of the melting temperatures (Tm s) were calculated to assess the reliability, and the limit of detection was examined to assess the sensitivity. RESULTS: All genotypes were correctly identified with the new method, and its accuracy was higher than that of fragment analysis. The intra- and inter-run coefficients of variation for the Tm s were both ≤0.27%, with standard deviations ≤0.14°C. The limit of detection was 0.2 ng of input genomic DNA. CONCLUSION: The developed PCR melting curve analysis using one fluorescent probe can provide accurate, reliable, rapid, simple, and low-cost detection of UGT1A1 (TA)n polymorphism, and its use can be easily generalized in clinical laboratories with a fluorescent PCR platform.

Persistent increase of carbohydrate antigen 19-9 with an unknown reason: A seven-year follow-up case.

BACKGROUND: We described a patient who exhibited a gradual increase in carbohydrate antigen 19-9 (CA19-9) concentrations for 4 years at three hospitals, with no associated clinical manifestations; however, we were unable to define the cause of this increase, forcing us to consider whether it was a false-positive result. METHODS: Given the potential for interference, this study used multiple system detection, gradient dilution, Polyethylene glycol (PEG) precipitation and heterophilic antibody blocking assay to evaluate the reliability of CA19-9 concentration increase. RESULTS: Analysis of the patient sample using multiple systems indicated that CA19-9 concentrations showed an obvious increase (154.0, and 889.2 IU/ml, respectively) using the Cobas E602 and Advia Centaur XP systems, and were within the reference ranges (<10 IU/ml) on other modules. PEG precipitation on the Cobas E602 and Advia Centaur XP systems reduced the CA19-9 concentration, as did heterophilic blocking tube (HBT-6, HBT-1) blockade. CONCLUSION: CA19-9 was incorrectly identified to increase due to the presence of heterophilic antibodies. We recommend that heterophilic antibodies should be evaluated in cases with elevated CA19-9 level but no associated clinical manifestations to prevent false positives.

[Phenotypes and genotypes of 78 patients with propionic acidemia].

Objective: Propionic acidemia is a rare inherited metabolic disorder caused by propionyl CoA carboxylase (PCC) deficiency. This study aims to analyze the clinical characteristics and gene variations of Chinese patients with propionic acidemia, and to explore the correlation between clinical phenotypes and genotypes. Methods: Single-center, retrospective and observational study. Seventy-eight patients of propionic acidemia (46 males and 32 females) from 20 provinces and autonomous regions were admitted from January 2007 to April 2022. Their age of initial diagnosis ranged from 7 days to 15 years. The clinical manifestations, biochemical and metabolic abnormalities, genetic variations, diagnosis, treatment and outcome were studied. Chi-Square test or Mann-Whitney U test were used for statistical analysis. Results: Among 78 cases, 6 (7.7%) were identified by newborn screening; 72 (92.3%) were clinically diagnosed after onset, and the age of onset was 2 hours after birth to 15 years old; 32 cases had early-onset disease and 40 cases had late-onset disease. The initial manifestations included lethargy, hypotonia, vomiting, feeding difficulties, developmental delay, epilepsy, and coma. Among the 74 cases who accepted gene analysis, 35 (47.3%) had PCCA variants and 39 (52.7%) had PCCB variants. A total of 39 PCCA variants and 32 PCCB variants were detected, among which c.2002G>A and c.229C>T in PCCA and c.838dupC and c.1087T>C in PCCB were the most common variants in this cohort. The variants c.1228C>T and c.1283C>T in PCCB may be related to early-onset type. The variants c.838dupC, c.1127G>T and c.1316A>G in PCCB, and c.2002G>A in PCCA may be related to late-onset disease. Six patients detected by newborn screening and treated at asymptomatic stage developed normal. The clinically diagnosed 72 cases had varied complications. 10 (12.8%) cases of them died. 62 patients improved after metabolic therapy by L-carnitine and diet. Six patients received liver transplantation because of recurrent metabolic crisis. Their clinical symptoms were markedly improved. Conclusion: The clinical manifestations of propionic acidemia are complex and lack of specificity. Newborn screening and high-risk screening are keys for early treatment and better outcome. The correlation between the genotype and phenotype of propionic acidemia is unclear, but certain variants may be associated with early-onset or late-onset propionic acidemia.

Subgroup analysis of the influence of body mass index on the association between serum lipids and cognitive function in Chinese population.

BACKGROUND: Previous studies reported that the association between lipid levels and cognitive function is related with gender, age and specific cognitive domains, but the influence of body mass index (BMI) on this association is limited. This triggered interest in exploring how serum lipids relate to cognitive function in different subgroups. METHODS: Data was collected from 2009 wave and 2015 wave of China Health and Nutrition Survey (CHNS). Multivariable linear regression analyses examined serum lipids level as predictors of sex- and age-specific measure of cognitive function in different BMI levels, which were adjusted for nationality, BMI, systolic blood pressure (SBP), diastolic blood pressure (DBP), smoking status, alcohol consumption and education level. RESULTS: Cognitive function score have different concentration curves in serum lipids quartile levels in different BMI categories. After adjustment for confounding factors, serum TG was positively associated with cognitive function score in underweight (ß ± SE: 2.06 ± 0.88, P = 0.023) and obese (ß ± SE: 1.44 ± 0.71, P = 0.045) male group, and serum HDL-C was positively associated with cognitive function score in overweight (ß ± SE: 1.89 ± 0.92, P = 0.041) and obese (ß ± SE: 5.04 ± 1.62, P = 0.002) female group. Serum TC was negatively associated with cognitive function score in overweight (ß ± SE: - 2.55 ± 1.26, P = 0.043) mid-life adults, and serum HDL-C was positively associated with cognitive function score in overweight (ß ± SE: 2.15 ± 0.94, P = 0.022) and obese (ß ± SE: 5.33 ± 2.07, P = 0.011) older adults. CONCLUSION: The associations of serum lipids with cognitive function were related with BMI levels and differed between gender and age groups. This result indicated that better nutritional status has superior cognitive function performance.

Short-term biological variation and Reference change values of urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine and 8-oxo-7,8-dihydroguanosine.

AIM: The DNA and RNA oxidative damage products urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-odGsn) and 8-oxo-7,8-dihydroguanosine (8-oGsn) have potential use in clinical practice. However, biological variation (BV) and reference change values (RCVs) have not been established. The aim of this study was to establish the short-term between-subject BV(CVG ), within-subject BV(CVI ), and RCVs for urinary 8-odGsn and 8-oGsn. METHODS: First-morning midstream urine specimens were collected from 20 apparently healthy subjects(ten males and ten females) on five consecutive days. 8-odGsn and 8-oGsn were measured using LC-MS/MS, while urine creatinine (U-Cr) was also measured to correct their results. A two-level nested ANOVA was used to estimate the CVI and CVG. RESULTS: The values of CVG for 8-odGsn, 8-odGsn/U-Cr, 8-oGsn, and 8-oGsn/U-Cr were 31.2%, 39.6%, 35.3%, and 28.8%, respectively, while CVI for them were 40.5%, 9.0%, 33.5%, and 12.1%, respectively. The RCVs for 8-odGsn, 8-odGsn/U-Cr, 8-oGsn, and 8-oGsn/U-Cr were 112.5%, 26.7%, 93.7%, and 36.5%, respectively. CONCLUSION: BV and RCVs were firstly established for 8-oxo-dGsn and 8-oGsn, and can be used in clinical practice.

Associations of urinary polycyclic aromatic hydrocarbons with albuminuria in U.S. adults, NHANES 2003-2014.

BACKGROUND: Polycyclic aromatic hydrocarbons (PAHs) exposure has been shown to be a risk factor for many diseases. However, studies on the association between PAHs exposure and kidney disease are limited. The aim of this study was to explore the association between urinary PAHs and albuminuria based on a national representative sample from the general U.S. METHOD: The data utilized were extracted from the 2003-2014 National Health and Nutrition Examination Survey (NHANES). Eight urinary PAHs were detected as PAH metabolites (OH-PAHs). Multivariable logistic regression analyses were applied to examine the association between urinary OH-PAHs and urinary albumin-creatinine ratio (ACR). All models were adjusted for confounding demographic, anthropometric and lifestyle factors. RESULT: A total of 8149 NHANES (2003-2014) participants with complete data were eligible. Compared with the lowest quartile, an increased prevalence of high ACR level (>3 mg/mmol) was observed in the participants with the highest quartile of 2-hydroxynaphthalene [OR (95% CI), 1.56 (1.28-1.90), P < 0.001], 3-hydroxyfluorene [OR (95% CI), 1.29 (1.06-1.58), P = 0.011] and 2-hydroxyfluorene [OR (95% CI), 1.47 (1.20-1.80), P < 0.001] levels after adjusting for confounding factors. In subgroup analysis, significantly high OH-PAHs leveland a strong relationship between OH-PAHs and ACR were observed in current smokers in the adjusted model. CONCLUSION: High levels of urinary OH-PAHs were positively associated with high levels of ACR in the U.S. POPULATION: Our finding provided evidence that PAHs exposure might potentially be related to albuminuria and therefore might have implications for environmental governance and prevention/treatment of this condition.

Association of body mass index and blood lipid profile with cognitive function in Chinese elderly population based on data from the China Health and Nutrition Survey, 2009-2015.

AIM: The associations of body mass index (BMI) and serum lipids with cognitive function are inconsistent and remain unclear, especially in the elderly population. This discrepancy triggered our interest in exploring the impact of BMI and serum lipids on memory status among the elderly Chinese population. METHODS: Data were collected from the China Health and Nutrition Survey database. We used data from the survey's 2015 wave to examine the association between BMI and memory status and from the 2009-2015 surveys to examine the association between serum lipids and cognitive function. We performed multivariable logistic regression analyses and multivariable linear regression analyses to examine these associations. RESULTS: Being underweight, normal weight, and severely obese were associated with an increased risk of bad self-reported memory status, with overweight as the reference. After adjustment for confounding factors, BMI was positively associated with cognitive function score in the low BMI group (≤24.5 kg/m2 ) (ß ± SE: 0.02 ± 0.01, P = 0.013) and negatively associated with cognitive function score in the high BMI group (>24.5 kg/m2 ) (ß ± SE: -0.04 ± 0.01, P = 0.009) in multivariable linear regression analysis. In men, higher levels of serum triglycerides and apolipoprotein B were associated with a decreased risk of cognitive impairment. In women, a higher level of high-density lipoprotein cholesterol was associated with a decreased risk of cognitive impairment. CONCLUSION: We found inverse U-shaped relationships between BMI and cognitive function and for the gender-specific association of serum lipids with cognitive function. This result indicated that among the elderly population, better nutritional status suggests superior memory status and cognitive function performance.

Genetically elevated circulating homocysteine concentrations increase the risk of diabetic kidney disease in Chinese diabetic patients.

Diabetic kidney disease (DKD) is a devastating and frequent complication of diabetes mellitus. Here, we first adopted methylenetetrahytrofolate reductase (MTHFR) gene C677T polymorphism as an instrument to infer the possible causal relevance between circulating homocysteine and DKD risk in a Chinese population and next attempted to build a risk prediction model for DKD. This is a hospital-based case-control association study. Total 1107 study participants were diagnosed with type 2 diabetes mellitus, including 547 patients with newly diagnosed and histologically confirmed DKD. MTHFR gene C677T polymorphism was determined using the TaqMan method. Carriers of 677TT genotype (14.55 µmol/L) had significantly higher homocysteine concentrations than carriers of 677CT genotype (12.88 µmol/L) (P < 0.001). Carriers of 677TT genotype had a 1.57-fold increased risk of DKD (odds ratio: 1.57, 95% CI: 1.21-2.05, P = 0.001) relative to carriers of 677CT genotype after adjusting for confounders. Mendelian randomization analysis revealed that the odds ratio for DKD relative to diabetes mellitus per 5 µmol/L increment of circulating homocysteine concentrations was 3.86 (95% confidence interval: 1.21-2.05, P < 0.001). In the Logistic regression analysis, hypertension, homocysteine and triglyceride were significantly associated with an increased risk of DKD and they constituted a risk prediction model with good test performance and discriminatory capacity. Taken together, our findings provide evidence that elevated circulating homocysteine concentrations were causally associated with an increased risk of DKD in Chinese diabetic patients.

Interaction of MTHFR C677T polymorphism with smoking in susceptibility to diabetic nephropathy in Chinese men with type 2 diabetes.

We investigated the interaction of MTHFR C677T polymorphism (rs1801133) with smoking in susceptibility to diabetic nephropathy (DN) in Chinese men with type 2 diabetes mellitus (T2DM). We studied 655 Chinese men with T2DM, who were divided into two groups (321 with DN and 334 without DN). The genotype of MTHFR C677T polymorphism was detected by real-time polymerase chain reaction. MTHFR TT genotype carried a higher risk of DN compared with the CC genotype (OR = 2.05; P = 0.002). The T allele showed marked association with DN development in patients who smoked, using additive, recessive, and dominant models (OR = 1.60, 1.83, and 1.88, respectively; P = 0.006, 0.002, and 0.04, respectively), which was not observed in the nonsmoking group. Patients with TT and CT genotypes, who smoked had a higher risk of DN compared with the control group (non-smoking with CC genotype; OR = 3.73 and 2.28, respectively; P < 0.001 and P = 0.004, respectively), whereas the other groups were not observed. In conclusion, the T allele of rs1801133 may be a risk factor for DN in Chinese men with T2DM, and synergy appears to exist between the MTHFR rs1801133 and smoking in susceptibility to DN.

Characteristics of women infected with human papillomavirus in a tertiary hospital in Beijing China, 2014-2018.

BACKGROUND: Human papillomavirus (HPV) infection is the main cause of precancerous lesions and cervical cancer in women. In order to determine the epidemiological characteristics as well as the relationship between the HPV genotype and cytology test results among women in Beijing, China, we retrospectively collected and analyzed the data from a tertiary hospital in Beijing, China. METHODS: A total of 21,239 women visited the China-Japan Friendship Hospital between 2014 and 2018 and their cervical exfoliations were collected. Thirteen HPV subtypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68) were examined and ThinPrep cytological test (TCT) was performed. RESULTS: Among all cases, 4473 (21.06%) women were infected with HPV. HPV52 (4.64%), HPV16 (4.44%) and HPV58 (4.28%) had higher prevalence. Single-type infection (77.26%) was more common than multiple-type infection (22.74%). Single-type infection was more frequently seen in women aged 50-60 years (17.17%), and multiple-type infection was more common in those aged < 30 years (7.88%). Significant differences in secular trends from 2014 to 2018 were observed for subtypes HPV39, 51, 52 and 58. HPV positive rates of women aged < 30 and 30-40 years changed significantly along with the time period, and the TCT positive rates of women aged 30-40, 40-50, 50-60 and >  60 years also showed significant differences from 2014 to 2018. In addition, 1746 (8.22%) women were TCT positive, of whom, 858 (4.04%), 561 (2.64%) and 327 (1.54%) had atypical squamous cells (ASCs), low-grade squamous intraepithelial lesions (LSILs) and high-grade squamous intraepithelial lesions (HSILs), respectively. Among four types of cytological lesions, the HPV infection rates were 16.76, 66.08, 63.99 and 85.32% in those negative for intraepithelial lesions or malignancy (NILM), ASC, LSIL and HSIL, respectively. CONCLUSIONS: HPV52, 16 and 58 are the most common infection subtypes in this study and among four types of cytological lesions, HSILs has the highest HPV prevalence. Significant differences in secular trends are observed for different subtypes in recent 5 years. The results on HPV genotype-specific prevalence should be considered when the HPV vaccine program is implemented in Beijing area.

Serum levels of IL-17 are elevated in patients with acute gouty arthritis.

Acute gouty arthritis (AGA) is one of the most common forms of auto-inflammatory arthritis. IL-17 is a key proinflammatory cytokine which has been implicated in several autoimmune diseases. However, to date little is known about the role of IL-17 in AGA. In the present study, we show that serum IL-17 levels are significantly elevated in AGA patients early in the onset of symptoms of gout, and decrease gradually as symptoms diminish. Correlation analysis indicated that IL-17 expression is not only positively correlated with disease activity, but is also correlated with serum levels of IL-1ß which plays a critical role in the differentiation of IL-17- γδT cells into IL-17+γδT cells. Flow cytometry analysis indicated that γδ T cells are a major source of IL-17 production during the early onset of AGA. We therefore identify IL-17 as a potential novel biomarker for AGA and suggest that targeting the γδ T cell/IL-17 immune axis is a potential strategy for treatment of acute flares of AGA.

Nosocomial transmission of Clostridium difficile ribotype 027 in a Chinese hospital, 2012-2014, traced by whole genome sequencing.

BACKGROUND: The rapid spread of Clostridium difficile NAP1/BI/027 (C. difficile 027) has become one of the leading threats of healthcare-associated infections worldwide. However, C. difficile 027 infections have been rarely reported in Asia, particularly in China. RESULTS: In this study, we identified a rare C. difficile bloodstream infection (BSI) from three isolates of a patient during repeated hospital admission. This finding triggered a retrospective epidemiological study to scan all cases and strains emerged from this ward during the past three years. Using medical personnel interviews, medical record reviews and the genomic epidemiology, two outbreaks in 2012 and 2013-2014 were identified. Through using whole genome sequencing, we succeeded to trace the origin of the BSI strain. Surprisingly, we found the genome sequences were similar to C. difficile 027 strain R20291, indicating the occurrence of a rare C. difficile 027 strain in China. Integrated epidemiological investigation and whole genome sequencing of all strains, we constructed a nosocomial transmission map of these two C. difficile 027 outbreaks and traced the origin of the infection. CONCLUSIONS: By genome sequencing, spatio-temporal analysis and field epidemiology investigation, we can estimate their complex transform network and reveal the possible modes of transmission in this ward. Based on their genetic diversity, we can assume that the toilets, bathroom, and janitor's equipment room may be contaminated area, which may be suggested to improve infection control measures in the following health care.

Detection the Frequency and Characteristics of FLT3 Internal Tandem Duplication Mutations by Capillary Electrophoresis Assay and Next-Generation Sequencing in.

BACKGROUND: FLT3-internal tandem duplication mutations (ITDs) are found in approximately 30% of patients with acute myeloid leukemia (AML) and are markers of poor prognosis. However, the characteristics of FLT3/ ITDs in Chinese AML patients have rarely been reported. The aim of this study was to analyze the frequency and characteristics of FLT3/ITDs in Chinese AML patients. METHODS: In the selected 152 cases of Chinese AML patients, capillary electrophoresis (CE) was used to analyze the frequency and characteristics of FLT3/ITDs. Next-generation sequencing was used to analyze the sequences of FLT3/ITDs positive patients. The differences of clinical features between FLT3/ITD positive group and FLT3/ITD negative group were estimated by statistical analysis. RESULTS: 42 cases (27.6%) were FLT3/ITDs-positive, in which 34 cases (81%) had a single duplication, and the remaining 8 cases (19%) had 2 or more ITDs. Median ITD size was 42 bp and median ITD allelic ratio was 0.25. Using next-generation sequencing, ITDs integrating in non-juxtamembrane (JM) domains were detected in 12 ITDs (24%) and in JM domains were detected in 38 ITDs (76%). Furthermore, duplication of at least one residue between Y591 and Y599 was detected in 48 (96%) of all 50 ITDs, and the insertion site was strongly correlated with ITD size: more C-terminal located inserted fragments were significantly longer. CONCLUSIONS: Our data provide further evidence of the heterogeneity of FLT3/ITDs among different subgroups in Chinese AML patients. ITDs varied widely, but hotspots were concentrated. These results also suggest that nextgeneration sequencing is a useful method for detection of FLT3/ITDs sequences.

Regulation of soluble epoxide hydrolase in renal-associated diseases: insights from potential mechanisms to clinical researches.

In recent years, numerous experimental studies have underscored the pivotal role of soluble epoxide hydrolase (sEH) in renal diseases, demonstrating the reno-protective effects of sEH inhibitors. The nexus between sEH and renal-associated diseases has garnered escalating attention. This review endeavors to elucidate the potential molecular mechanisms of sEH in renal diseases and emphasize the critical role of sEH inhibitors as a prospective treatment modality. Initially, we expound upon the correlation between sEH and Epoxyeicosatrienoic acids (EETs) and also addressing the impact of sEH on other epoxy fatty acids, delineate prevalent EPHX2 single nucleotide polymorphisms (SNPs) associated with renal diseases, and delve into sEH-mediated potential mechanisms, encompassing oxidative stress, inflammation, ER stress, and autophagy. Subsequently, we delineate clinical research pertaining to sEH inhibition or co-inhibition of sEH with other inhibitors for the regulation of renal-associated diseases, covering conditions such as acute kidney injury, chronic kidney diseases, diabetic nephropathy, and hypertension-induced renal injury. Our objective is to validate the potential role of sEH inhibitors in the treatment of renal injuries. We contend that a comprehensive comprehension of the salient attributes of sEH, coupled with insights from clinical experiments, provides invaluable guidance for clinicians and presents promising therapeutic avenues for patients suffering from renal diseases.

Serum circPRDM5 as a novel diagnostic biomarker for acute myocardial infarction.

BACKGROUND: Circular RNA (CircRNA) is known to play an important role in cardiovascular diseases, but its use as a biomarker of acute myocardial infarction (AMI) has not been studied. This study explores the feasibility of circPRDM5 as a novel biomarker of AMI. METHODS: CircPRDM5 was screened by bioinformatics, the correct circPRDM5 primers were tested by agarose gel electrophoresis (AGE) and Sanger sequencing, and the expression level of serum circPRDM5 was detected by Quantitative Reverse Transcription-Polymerase Chain Reaction. (qRT-PCR), and the diagnostic value of circPRDM5 was analyzed by the receiver operating characteristic (ROC) curve. RESULTS: The expression of circPRDM5 in serum of AMI patients was significantly decreased compared with that of healthy control group and angina group (P < 0.001). The area under ROC curve of serum circPRDM5 was 0.862 [95 % CI, 0.814-0.909]. The combined diagnosis of serum circPRDM5, cardiac troponin T (cTnT) and creatine kinase-MB (CK-MB) could improve the sensitivity of diagnosing AMI. The expression level of serum circPRDM5 increased after percutaneous coronary intervention (PCI). CONCLUSIONS: CircPRDM5 can be used as a novel biomarker for AMI, and its combination with cTnT and CK-MB can improve diagnostic value.