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Background To study the association of beta-carotene intake from diet with anxiety in US midlife women. Methods Analyses were performed on the baseline data of the Study of Women's Health Across the Nation (SWAN), a multicenter and population-based study of the natural history of US midlife women aged from 42 to 52. Logistic regression and restricted cubic spline models were performed to examine the association of beta-carotene intake with anxiety. Results A total of 3051 midlife women were included in the present study. In early perimenopausal women, the odds ratios (ORs) of anxiety in the crude model indicated that beta-carotene intake was inversely associated with anxiety. After additional adjustment for age, race/ethnicity, education, income, financial strain, physical activity, BMI, vasomotor symptoms (VMS), chronic stress, use of antidepressant and total caloric intake in model 1, the results were similar to those of the crude model. This association remained statistically significant and changed little when additional controlling for estradiol, testosterone and sex hormone binding globulin (SHBG) in the fully adjusted model 2. The fully adjusted ORs and 95% confidence intervals (CIs) were 0.606 (0.408-0.901). However, in premenopausal women, no statistically significant difference was observed between beta-carotene intake and anxiety. Limitations This was a cross-sectional study, limiting causal inferences. Conclusion Dietary beta-carotene intake may be inversely associated with anxiety in early perimenopausal women, but not in premenopausal women.

Background In December 2019, the coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) broke out in Wuhan. Epidemiological and clinical characteristics of patients with COVID-19 have been reported, but the relationships between laboratory features and viral load has not been comprehensively described. Methods Adult inpatients (≥18 years old) with COVID-19 who underwent multiple (≥5 times) nucleic acid tests with nasal and pharyngeal swabs were recruited from Renmin Hospital of Wuhan University, including general patients (n = 70), severe patients (n = 195), and critical patients (n = 43). Laboratory data, demographic data, and clinical data were extracted from electronic medical records. The fitted polynomial curve was used to explore the association between serial viral loads and illness severity. Results Viral load of SARS-CoV-2 peaked within the first few days (2–4 days) after admission, then decreased rapidly along with virus rebound under treatment. Critical patients had the highest viral loads, in contrast to the general patients showing the lowest viral loads. The viral loads were higher in sputum compared with nasal and pharyngeal swab (P = .026). The positive rate of respiratory tract samples was significantly higher than that of gastrointestinal tract samples (P < .001). The SARS-CoV-2 viral load was negatively correlated with portion parameters of blood routine and lymphocyte subsets and was positively associated with laboratory features of cardiovascular system. Conclusions The serial viral loads of patients revealed whole viral shedding during hospitalization and the resurgence of virus during the treatment, which could be used for early warning of illness severity, thus improve antiviral interventions.

Background To study the association of trans fatty acid intake from diet with depressive symptoms in midlife women. Methods Data for this cross-sectional study were retrieved from baseline assessment of the Study of Women's Health Across the Nation (SWAN). Linear regression, logistic regression and restricted cubic spline models were performed to examine the association trans fatty acid intake with depressive symptoms. Results A total of 3095 women aged 42–52 years were included in the present study. Trans fatty acid intake was positively associated with CES-D (Center for Epidemiological Studies-Depression) scores in unadjusted, age-, race/ethnicity-, total family income- and education-adjusted and fully adjusted linear regression model. The fully adjusted regression coefficient with 95% confidence intervals (CIs) of trans fatty acid intake were 0.146 (0.047-0.246) for CES-D score. Trans fatty acid intake was positively associated with depressive symptoms (CES-D score ≥16) in each logistic regression model. The fully adjusted odds ratios (ORs) with 95% CI of depressive symptoms were 1.410 (1.019-1.951) in quartile 4 compared with quartile 1 for trans fatty acid intake. Sensitivity analyses showed that the association between trans fatty acid and depressive symptoms were not affected by use of antidepressant. Stratified analysis showed that, in fully adjusted model, the trans fatty acid intake was not significantly associated with depressive symptoms in early perimenopausal women. Limitations This was a cross-sectional study, limiting causal inferences. Conclusion Trans fatty acid may be positively associated with depressive symptoms in premenopausal women, but not in early perimenopausal women.

The outbreak and widely spread of coronavirus disease 2019 (COVID-19) has become a global public health concern. COVID-19 has caused an unprecedented and profound impact on the whole world, and the prevention and control of COVID-19 is a global public health challenge remains to be solved. The retrospective analysis of the large scale tests of SARS-CoV-2 RNA may indicate some important information of this pandemic. We selected 12400 SARS-CoV-2 tests detected in Wuhan in the first semester of 2020 and made a systematic analysis of them, in order to find some beneficial clue for the consistent prevention and control of COVID-19.SARS-CoV-2 RNA was detected in suspected COVID-19 patients with real-time fluorescence quantitative PCR (RT-qPCR). The patients' features including gender, age, type of specimen, source of patients, and the dynamic changes of the clinical symptoms were recorded and statistically analyzed. Quantitative and qualitive statistical analysis were carried out after laboratory detection.The positive rate of SARS-CoV-2 was 33.02% in 12,400 suspected patients' specimens in Wuhan at the first months of COVID-19 epidemics. SARS-CoV-2 RT-qPCR test of nasopharyngeal swabs might produce 4.79% (594/12400) presumptive results. The positive rate of SARS-CoV-2 RNA was significantly different between gender, age, type of specimen, source of patients, respectively (P < .05). The median window period from the occurrence of clinical symptom or close contact with COVID-19 patient to the first detection of positive PCR was 2 days (interquartile range, 1-4 days). The median interval time from the first SARS-CoV-2 positive to the turning negative was 14 days (interquartile range, 8-19.25 days).This study reveals the comprehensive characteristics of the SARS-CoV-2 RNA detection from multiple perspectives, and it provides important clues and may also supply useful suggestions for future work of the prevention and treatment of COVID-19.

Background The SARS-CoV-2 RNA was detected positive again after discharged from hospital in some COVID-19 patients, with or without clinical symptoms such as fever or dry cough. Methods 1008 severe COVID-19 patients, with SARS-CoV-2 RNA positive detected with the mixed specimen of nasopharyngeal swab and oropharyngeal swab by real-time fluorescence quantitative PCR (RT-qPCR), were selected to monitor SARS-CoV-2 RNA with the 12 types of specimens by RT-qPCR during hospitalization. All of 20 discharged cases with COVID-19 were selected to detect SARS-CoV-2 RNA in isolation period with 7 types of specimens by RT-qPCR before releasing the isolation period. Results Of the enrolled 1008 severe patients, the nasopharyngeal swab specimens showed the highest positive rate of SARS-CoV-2 RNA (71.06%), followed by alveolar lavage fluid (66.67%), oropharyngeal swab (30.77%), sputum (28.53%), urine (16.30%), blood (12.5%), stool (12.21%), anal swab (11.22%) and corneal secretion (2.99%), and SARS-CoV-2 RNA couldn’t be detected in other types of specimen in this study. Of the 20 discharged cases during the isolation period, the positive rate of SARS-CoV-2 RNA was 30% (6/20): 2 cases were positive in sputum at the eighth and ninth day after discharge, respectively, 1 case was positive in nasopharynx swab at the sixth day after discharge, 1 case was positive in anal swab at the eighth day after discharge, and 1 case was positive in 3 specimens (nasopharynx swab, oropharynx swab and sputum) simultaneously at the fourth day after discharge, and no positive SARS-CoV-2 RNA was detected in other specimens including stool, urine and blood at the discharged patients. Conclusions SARS-CoV-2 RNA should be detected in multiple specimens, such as nasopharynx swab, oropharynx swab, sputum, and if necessary, stool and anal swab specimens should be performed simultaneously at discharge when the patients were considered for clinical cure and before releasing the isolation period.

Background Glucose-6-phosphate dehydrogenase deficiency (D-G6PD) is an X-linked recessive disorder resulted from deleterious variants in the housekeeping gene Glucose-6-phosphate 1-dehydrogenase (G6PD), causing impaired response to oxidizing agents. Screening for new variations of the gene helps with early diagnosis of D-G6PD resulting in a reduction of disease related complications and ultimately increased life expectancy of the patients. Methods One thousand five hundred sixty-five infants with pathological jaundice were screened for G6PD variants by Sanger sequencing all of the 13 exons, and the junctions of exons and introns of the G6PD gene. Results We detected G6PD variants in 439 (28.1%) of the 1565 infants with pathological jaundice. In total, 9 types of G6PD variants were identified in our cohort; and a novel G6PD missense variant c.1118 T > C, p.Phe373Ser in exon 9 of the G6PD gene was detected in three families. Infants with this novel variant showed decreased activity of G6PD, severe anemia, and pathological jaundice, consistent with Class I G6PD deleterious variants. Analysis of the resulting protein’s structure revealed this novel variant affects G6PD protein stability, which could be responsible for the pathogenesis of D-G6PD in these patients. Conclusions High rates of G6PD variants were detected in infants with pathological jaundice, and a novel Class I G6PD deleterious variants was identified in our cohort. Our data reveal that variant analysis is helpful for the diagnosis of D-G6PD in patients, and also for the expansion of the spectrum of known G6PD variants used for carrier detection and prenatal diagnosis.

Protein phosphatase 2A (PP2A) was reported to play an important role in cancer development; however, the relationship between PP2A and cervical cancer development has yet to be fully understood. The present study aimed to explore the role of PP2A in the development of cervical cancer. Serum levels of PP2A were detected by ELISA in 23 patients with cervical cancer and 30 patients with benign cervical lesions. Furthermore, the PP2A activities and the mRNA and protein levels of PP2A were measured in cervical cancer (n=8) and chronic cervicitis (n=10) tissues. The results showed that the serum levels of PP2A were significantly reduced in patients with cervical cancer. Further studies showed that not only the activities of PP2A but also the mRNA and protein levels of PP2A were significantly decreased in cervical cancer tissues. Wound healing and Transwell assays demonstrated that pharmacological and genetic upregulation of PP2A could inhibit the migration of HeLa cells, but the downregulation of PP2A promoted cellular migration. The activation of PP2A also inhibited the remodeling of actin and the activity of mitogen-activated protein kinases (MAPKs) including p-JNK, p-p38 and p-ERK. Meanwhile, the activation of PP2A was found to downregulate MMP-9 levels, which further inhibited the migration and invasion of HeLa cells. In conclusion, our data suggest that the activity and expression of PP2A are significantly reduced in cervical cancer tissues, and the activation of PP2A may inhibit the migration of cervical cancer cells by inhibiting the phosphorylation of p-JNK, p-p38 and the p-ERK/MAPK signaling pathway as well as by downregulating MMP-9, implying that PP2A plays an important role in cervical cancer development.

Background To study the association of manganese intake from diet with depressive symptoms in midlife women. Methods Data for this cross-sectional study were retrieved from baseline assessment of the Study of Women's Health Across the Nation (SWAN). Linear regression, logistic regression and restricted cubic spline models were performed to examine the association of manganese intake with depressive symptoms. Results A total of 2993 midlife women aged 42–52 years were included in the present study. In premenopausal women, manganese intake was inversely associated with CES-D score and after adjustment for total caloric intake, age, race/ethnicity, education, income, financial strain, physical activity, BMI, vasomotor symptoms, chronic stress and use of antidepressant, estradiol, testosterone and sex hormone binding globulin using linear regression. The fully adjusted regression coefficient 95% confidence intervals (CIs) were −0.533 (−0.993, −0.074). Similarly, manganese intake was inversely associated with depressive symptoms (CES-D scores≥16) using logistic regression adjusted same confounders. The fully adjusted odds ratios (ORs) with 95% CI of depressive symptoms were 0.512 (0.287–0.913) in quartile 4 compared with quartile 1 for manganese intake. However, in early perimenopausal women, no statistically significant difference was observed between manganese intake and depressive symptoms after adjustment for same confounders that adjusted in premenopausal women. Limitations This was a cross-sectional study, limiting causal inferences. Assessment of CES-D was based on a self-report scale. Conclusion Manganese intake may be inversely associated with depression symptoms in premenopausal women, but not in early perimenopausal women.

Background This prospective investigation examined relations between baseline dietary saturated fatty acid (SFA) intake of women at midlife and prevalence of depressive symptoms 4 years later. Methods Women enrolled in the Study of Women's Health Across the Nation (SWAN) had measures of CES-D depression scores 4 years later and SFA intake at baseline. Logistic regression models were used to study prospective associations between SFA intake and depressive symptoms. Results A total of 2400 women aged 42–52 years were included in the present study. The ORs of depressive symptoms in model 1 adjustment for baseline CES-D score indicated that baseline SFA intake was positively associated with depressive symptoms. After additional adjustment for age, race/ethnicity, education, financial strain, physical activity, smoking currently, BMI, menopausal status, VMS, chronic stress and use of antidepressant in model 2, the results were similar to those of the model 1. This association remained statistically significant and changed little when additional controlling for SHBG and FSH in fully adjusted model 3. The fully adjusted OR of depressive symptoms in model 3 was 2.423 (1.142–5.143) in quartile 4 (the highest) versus quartile 1 (the lowest) of the baseline SFA intake. Limitations Assessment of depression and SFA intake was based on a self-report scale. Conclusions Baseline SFA intake is a predictor for depressive symptoms assessed 4 years later in midlife women. This underlines the importance of targeting SFA intake in the prevention of major depression in midlife women.

Background: This study aimed to evaluate the performance of modified carbapenem inactivation method (mCIM) combined EDTA-carbapenem inactivation method (eCIM), and inhibitor-based combined disk test (CDT) in the detection and distinguishing of carbapenemase production in Enterobacteriaceae. Methods: A total of 101 nonrepetitive carbapenem insensitive Enterobacteriaceae [minimal inhibitory concentration (MIC) ≥2 µg/mL] were tested by mCIM, eCIM and CDT respectively, and the major carbapenemase genes including blaKPC, blaNDM, blaIMP, blaVIM and blaOXA-48-like genes were detected by polymerase chain reaction (PCR) as control. Results: Seventy-nine (78.2%) of isolates were found to harbour one or more carbapenemase genes by PCR, with blaKPC and blaNDM being the most common genes. OXA-48-like genes were undetectable. The coincidence rate of mCIM combined eCIM and CDT was 97.5% (77/79) and 96.2% (76/79) respectively, compared with gene detection. Both assays had a misclassification in two bla KPC+NDM -producing isolates of Klebsiella oxytoca . The sensitivity and specificity of two assays above were 100.0% vs . 95.0% and 98.4% vs . 98.4%, respectively in distinguishing serine-carbapenemase, while they were 95.1% vs . 97.6% and 100% vs . 100.0%, respectively in distinguishing metallo-carbapenemase. Conclusions: mCIM combined eCIM and the CDT are both useful tools for the reliable detection and distinguishing single serine-carbapenemase or metallo-carbapenemase, but not for mixed types.

Background An inverse association between dietary fiber intake and depressive symptoms was reported in the general population, but this association is unstudied in midlife women. This study was designed to investigate the association of dietary fiber intake with depressive symptoms in midlife women. Methods Analyses for this cross-sectional study were performed on baseline assessment of the Study of Women's Health Across the Nation (SWAN). Linear regressions were used to examine the association of fiber intake with Center for Epidemiological Studies-Depression (CES-D) score. Logistic regression and restricted cubic spline analyses were used to examine the association between fiber intake and depressive symptoms (CES-D score ≥ 16). Results A total of 3054 midlife women in our study were stratified into premenopausal women and early perimenopausal women by menstrual bleeding patterns. In premenopausal women, dietary fiber intake was inversely associated with CES-D scores in unadjusted, age-, education-, race/ethnicity-, total family income-, BMI-, sport-, use of antidepressant-, dietary total caloric intake-, SHBG-, and FSH-adjusted linear regression model. The fully adjusted regression coefficient with 95% confidence intervals (CIs) of fiber intakes was -0.146 (-0.235, -0.058) for CES-D score. Fiber intake was inversely associated with depressive symptoms (CES-D score ≥ 16) in crude and fully adjusted logistic regression model. The fully adjusted odds ratios (ORs) with 95% CI of depressive symptoms was 0.483 (0.314-0.745) in quartile 4 compared with quartile 1 for fiber intake. However, in early perimenopausal women, dietary fiber intake was not statistically significantly associated with depressive symptoms. Conclusion Dietary fiber is inversely associated with depressive symptoms in premenopausal women, but not in early perimenopausal women.

Alport syndrome (AS) is a hereditary nephropathy characterized by glomerular basement membrane lesions. AS shows a relatively rare entity with autosomal dominant gene mutation (accounts for less than 5% of AS cases) and is widely believed to be a consequence of heterozygous variants in the COL4A3 and COL4A4 genes. Until now, there have been no reports of homozygous variants in genes in AS patients, and it is scarce to detect both homozygous and heterozygous variants in a single AS pedigree. We performed genetic analysis by exome sequencing (exome-seq) in a Chinese family with AS and found four individuals harboring the COL4A4 c.4599T > G variant, a novel COL4A4 nonsense mutation that gains stop codon and results in a truncated protein. The proband and her two siblings were determined to be heterozygous, whereas their mother was homozygous. The proband satisfied the criteria for the diagnosis of AS, which included clinical manifestations of microscopic hematuria and proteinuria, and pathological features of the glomerular basement membrane (GBM), including irregular thickening and splitting. However, the other three individuals who were homozygous or heterozygous for the variant exhibited mild clinical features with isolated microscopic hematuria. In summary, we identified a novel pathogenic variant in either the heterozygous or homozygous state of the COL4A4 gene in a Chinese family with AS. Our results also suggest that the severity of clinical manifestations may not be entirely attributed to by the COL4A4 genetic variant itself in patients.

Background : To study the association of n-3 polyunsaturated fatty acid (n-3 PUFA) intake from diet with depressive symptoms in midlife women. Methods : Data for this cross-sectional study were retrieved from baseline assessment of the Study of Women's Health Across the Nation (SWAN). Logistic regression and restricted cubic spline models were performed to examine the association n-3 PUFA intake with depressive symptoms. Results : A total of 3054 women aged 42–52 years were included in the present study. In overall midlife women, n-3 PUFA intake was not significantly associated with depressive symptoms (CES-D score ≥16) after adjustment for potential confounders. In early perimenopausal women, n-3 PUFA intake was inversely associated with depressive symptoms after adjustment for potential confounders. The fully adjusted OR with 95% CI of depressive symptoms were 0.062 (0.009–0.457) in the highest quartile (n-3 PUFAs intake was categorized by quartiles) compared with the lowest quartile for n-3 PUFA intake. Restricted cubic spline analyses showed that n-3 PUFA intake were dose–response inversely associated with depressive symptoms in early perimenopausal women. However, in premenopausal women, no significant association between depressive symptoms and n-3 PUFA intake was observed. Limitations : This was a cross-sectional study, limiting causal inferences. Conclusion : Dietary n-3 PUFA may be inversely associated with depressive symptoms in early perimenopausal women, but not in premenopausal women.

Objectives The aim of this study was to review the association of dietary intake of oleic and linoleic acids (OA and LA, respectively) with depressive symptoms in perimenopausal women. Methods This cross-sectional study used data from the Study of Women's Health Across the Nation (SWAN). Linear and logistic regressions and restricted cubic spline models were performed to examine the association of intake of OA and LA with depression. Results We included 2793 women 42 to 52 y of age in the present study. Intake of the two acids was positively associated with the Center for Epidemiologic Studies Depression Scale (CES-D) scores in unadjusted and age-, race/ethnicity-, total family income- and education-adjusted linear regression model. The fully adjusted regression coefficients were β = 0.089 and β = 0.145 for oleic and linoleic acid intake, respectively. OA and LA intake was positively associated with depressive symptoms (CES-D score ≥16) in unadjusted and age-, race/ethnicity-, total family income- and education-adjusted logistic regression model. The fully adjusted odds ratios (ORs) with 95% confidence intervals (CIs) of depressive symptoms were 1.994 (1.298–3.063) and 1.592 (1.047–2.421) for the highest versus lowest quartile of intake of OA and LA, respectively. Conclusion Intake of OA and LA may be positively associated with depressive symptoms in perimenopausal women.

The ageing process is accompanied by the gradual development of chronic systemic inflammation (inflamm-ageing). Growth differentiation factor 15 (GDF15) is associated with inflammation and known to be a stress-induced factor. The present study aimed to explore the association of GDF15 with ageing. In this cross-sectional study, serum GDF15, hematological parameters, and biomedical parameters were determined in 120 healthy individuals (23-83 years old, males). Three telomere related parameters, including telomere length, telomerase activity, and the expression of human telomerase reverse transcriptase (hTERT) mRNA were also quantified. Our results showed that the older group has a higher levels of GDF15 and lower expression of hTERT mRNA, and PBMC telomerase activity (p 0.001). In individuals with high GDF15 levels, they were older, and presented with the lower level of hTERT mRNA and T/S ratio (p 0.01). Spearman correlation analysis shows that GDF15 positively correlated with age (r = 0.664, p 0.001), and negatively correlated with telomere length (r = -0.434, p 0.001), telomerase activity (r = -0.231, p = 0.012), and hTERT mRNA (r = -0.206, p = 0.024). Furthermore, in multivariate regression analysis, GDF15 levels showed a statistically significant linear and negative relationship with PBMC telomerase activity (β-coefficient = -0.583, 95% CI -1.044 to -0.122, p = 0.014), telomere length (β-coefficient = -0.200, 95% CI -0.305 to -0.094, p 0.001), and hTERT mRNA (β-coefficient = -0.207, 95% CI -0.312 to -0.102, p 0.001) after adjusting for confounders. These results support that circulating GDF15 is the potential biomarker of ageing that may influence the risk and progression of multiple ageing conditions.

CNGA1 encodes a membrane protein on rod photoreceptor related to phototransduction. The present study was to identify a novel mutation of CNGA1 associated with autosomal recessive retinitis pigmentosa by using next generation sequencing of a Chinese family. Next generation sequencing and Sanger sequencing has identified a compound heterozygous mutation in CNGA1 gene, c0.472 del C (reported) and c0.829G>A (novel mutation, same as c0.622G>A according to NM_000087.3) of the proband. SIFT and Polyphen-2 predicted the CNGA1 G622A site to be possibly deleterious. Evolutionary conservation analysis of amino acid residues showed this aspartic acid is highly conserved between species, and protein structure prediction by I-TASSER server indicated that the D208N mutation induced a large disappear of interactions between S2 and S4. Flag-tagged CNGA1 and mutant G622A cDNA were generated and inserted into pCIG-eGFP vectors. Transfection of human embryonic kidney 293T cells was performed with lipofectamine. Interestingly, western blot and immunofluorescence results indicated that the expression of mutant CNGA1 (D208N) decreased significantly, especially on the membrane of transfected HEK293T cells. The novel variant c0.622G>A (p. D208N) in this study enriched the CNGA1 mutation spectrum. Besides, this mutant was predicted "possibly damaging" due to bioinformatics analysis and validated by laboratorial experiments. Our study suggests that this mutation lead to the CNGA1 protein reduction from the cell membrane.

Diagnosis of acute promyelocytic leukemia (APL) has been accelerated by multiparameter flow cytometry (MFC). However, diagnostic interpretation of MFC readouts for APL depends on individual experience and knowledge, which inevitably increases the risk of arbitrariness. We appraised the feasibility of using stepwise discriminant function analysis (SDFA) based on MFC to optimize the minimal variables needed to distinguish APL from other acute myeloid leukemia (AML) without complicated data interpretation. Samples from 327 patients with APL (n = 51) and non-APL AML (n = 276) were randomly allocated into training (243 AML) and test sets (84 AML) for SDFA. The discriminant functions from SDFA were examined by correct classification, and the final variables were validated by differential expression. Finally, additional 20 samples from patients with atypical APL and AML confusable with APL were also identified by SDFA method and morphological analysis. The weighed discriminant function reveals seven differentially expressed variables (CD2/CD9/CD11b/CD13/CD34/HLA-DR/CD117), which predict a molecular result for APL characterization with an accuracy that approaches 99% (99.6 and 98.8% for AML samples in training and test sets, respectively). Furthermore, the SDFA outperformed either single variable analysis or the more limited 3-component analysis (CD34/CD117/HLA-DR) via separate SDFA, and was also superior to morphological analysis in terms of diagnostic efficacy. The established SDFA based on MFC with seven variables can precisely and rapidly differentiate APL and non-APL AML, which may contribute to the urgent initiation of all-trans-retinoic acid-based APL therapy.

MiR-139-5p has been reported to be overexpressed in many types of cancers, but its role in bladder cancer has not been elucidated yet. Here, we report that miR-139-5p functions as a tumor suppressor in bladder cancer and inhibits the cancer stem cell self-renewal by targeting Bmi1 directly. We found that miR-139-5p expression was significantly downregulated in the bladder cancer specimens compared with that in adjacent normal tissues. In vitro, restoration of miR-139-5p expression significantly inhibited the proliferation of bladder cancer cells. Mechanism analysis revealed that miR-139-5p could decrease Bmi1 protein levels by binding to the 3′ untranslated region of Bmi1 messenger RNA. Stem cell–related proteins such as c-MYC, NANOG, OCT4, and KLF4 and signaling pathways such as Wnt signaling were suppressed by restoration of miR-139-5p in bladder cancer cells. In addition, miR-139-5p expression also blocked self-renewal of bladder cancer stem cells by inhibiting Bmi1. In summary, our study supports that miR-139-5p acts as a tumor suppressor in bladder cancer development and suppresses cancer stem cell property of bladder cancer. Our study also suggests that miR-139-5p has the potential to be used as a therapeutic molecule for bladder cancer treatment.

Approximately 170 million people in the world are infected with Hepatitis C virus (HCV). There are no published population based studies about the prevalence of HCV genotypes and the associations of genotype and Infection frequency with gender and age in WuHan. We aimed to investigate the distribution of HCV prevalence and genotypes among different gender and age patients with chronic HCV infection in WuHan from 2011 to 2015. A total of 2685 anti-HCV positive serum samples from individuals of physical examinationwere recruited from the Renmin Hospital of WuHan University, Hubei Province in China from January 2011 to December 2015. From these 2685 anti-HCV positive serum samples, 496 samples were with a positive PCR for HCV RNA. The number of HCV infection showed an increase with year, but the annual infection rate has remained similar (χ(2) = 2.94, P = 0.568). 2685 cases were infected with HCV from 2011 to 2015 in WuHan city of China. Blood transfusion (18.14 %) was the main routs of transmission, followed by Surgery (8.94 %). The highest prevalence of HCV infection was at the age group 50-59 (25.85 % of 2685) and the lowest prevalence was 0-9 (0.93 % of 2685). HCV genotype 1 was the most prevalent (73.39 %), followed by genotypes 2 (17.14 %), 3 (5.25 %) and 6 (3.22 %). Genotype 4 and 5 was not detected in these patients. The most prevalent subtype was subtype 1b (71.98 %), followed by genotypes 2a (17.14 %). Five patients had mixed infection across the HCV subtypes. Among all genotypes, genotype 1 was highest in both male (73.27 %) and female (73.47 %) patients, followed by genotype 2. Genotype 1 (male: 29.84 % of 496, vs female: 43.55 % of 496, χ(2) = 20.07, P 0.0001), genotype 2 (male: 6.25 % of 496, vs female: 10.89 % of 496, χ(2) = 6.81, P = 0.009), and 6 (male: 1.41 % of 496, vs female: 1.81 % of 496, χ(2) = 0.626, P = 0.401) were more common in female patients than males, while no significant gender differences were observed for genotype 6. Among age group 50-59 years Genotype 1 was most common in male patients (29.05 % of 148) followed in 20-29 years (23.65 % of 148), genotype 2 in the age group 60-69 (12 cases of 31) and genotype 3 in the age group 50-59 (4 cases of 13) and genotype 6 was most frequent in the age group 30-39 (4 cases of 7). The frequency of HCV prevalence was significantly higher in female patients compared to males before ages 60, while the opposite result was observed after 60 years. The most common HCV genotype in WuHan was subtype 1b followed by 2a and more common among women than males patients. Further studies are needed to collect a large number of samples to estimate the different epidemiology of the HCV genotypes, because the sample size of non-genotype 1b and 2a is not large enough and other factors like disease history/monthly income/etc. are not included in our study.

PKHD1 gene mutations are found responsible for autosomal recessive polycystic kidney disease (ARPKD). However, it is inconvenient to detect the mutations by common polymerase chain reaction (PCR) because the open reading frame of PKHD1 is very long. Recently, long-range (LR) PCR is demonstrated to be a more sensitive mutation screening method for PKHD1 by directly sequencing. In this study, the entire PKHD1 coding region was amplified by 29 reactions to avoid the specific PCR amplification of individual exons, which generated the size of 1 to 7 kb products by LR PCR. This method was compared to the screening method with standard direct sequencing of each individual exon of the gene by a reference laboratory in 15 patients with ARPKD. The results showed that a total of 37 genetic changes were detected with LR PCR sequencing, which included 33 variations identified by the reference laboratory with standard direct sequencing. LR PCR sequencing had 100% sensitivity, 96% specificity, and 97.0% accuracy, which were higher than those with standard direct sequencing method. In conclusion, LR PCR sequencing is a reliable method with high sensitivity, specificity and accuracy for detecting genetic variations. It also has more intronic coverage and lower cost, and is an applicable clinical method for complex genetic analyses.

Background ADH1B Arg48His polymorphism is associated with the development of alcohol-related diseases. In this study, we aimed to explore an improved polymerase chain reaction with confronting two-pair primers (PCR-CTPP) assay for the detection of ADH1B Arg48His polymorphism. Methods A mismatch was introduced at the 3′ end of each of the two allele-specific to increase the specificity of the reaction. But beyond that, a new mismatch at-3 positions of outer primers was designed to decrease the efficiency of the aforementioned primers and depresses the amplification of an internal nonspecific DNA control. A total of 180 samples from healthy volunteers Han Chinese were tested to evaluate this new assay. Results The protocol of PCR-CTPP was successful for genotyping of ADH1B Arg48His. The results from the improved PCR-CTPP assay were confirmed by Sanger sequencing, and correct genotyping rates were 100%.The genotype frequencies were 49.44% (89 cases) for His/His, 46.67% (84 cases) for Arg/His, and 3.89% (seven cases) for Arg/Arg respectively. Conclusions This improved PCR-CTPP assay is simple, rapid, cost-effective, and reliable, specific for the detection of ADH1B Arg48His polymorphism in most clinical diagnostic laboratories.

HBV (hepatitis B virus) genotyping is important in determining the clinical manifestation of disease and treatment response, particularly, in patients with low viral loads. Also, sensitive detection of HBV antiviral drug resistance mutations is essential for monitoring therapy response. Asensitive direct sequencing method for genotyping and the drug resistance mutation detection of low levels of HBV DNA in patients’ plasma is developed by PCR amplification of the DNA with novel universal primers. The novel, common, and universal primers were identified by alignment of RT region of all the HBV DNA sequences in databases. These primers could efficiently amplify the RT region of HBV virus at low DNA levels by directly sequencing the resulting PCR products, and mapping with the reference sequence made it possible to clearly obtain the HBV subtypes and identify the resistance mutations in the samples with HBV DNA level as low as 20 IU/mL. We examined the reliability of the method in clinical samples, and found it could detect the HBV subtypes and drug resistance mutations in 80 clinical HBV samples with low HBV DNA levels ranging from 20 to 200 IU/mL. This method is a sensitive and reliable direct sequencing method for HBV genotyping and antiviral drug resistance mutation detection, and is helpful for efficiently monitoring the response to therapy in HBV patients.