Correlation between the cardiometabolic index and arteriosclerosis in patients with type 2 diabetes mellitus.

BACKGROUND: The cardiometabolic index (CMI) is a new metric derived from the triglyceride-glucose index and body mass index and is considered a potential marker for cardiovascular risk assessment. This study aimed to examine the correlation between the CMI and the presence and severity of arteriosclerosis in patients with type 2 diabetes mellitus (T2DM). METHODS: This study involved 2243 patients with T2DM. The CMI was derived by dividing the triglyceride level (mmol/L) by the high-density lipoprotein level (mmol/L) and then multiplying the quotient by the waist-to-height ratio. Multivariate logistic regression was used to analyze the correlations between the CMI and BMI blood biomarkers, blood pressure, and brachial-ankle pulse wave velocity (baPWV). RESULTS: Patients were categorized into three groups based on their CMI: Group C1 (CMI < 0.775; n = 750), Group C2 (CMI: 0.775-1.355; n = 743), and Group C3 (CMI > 1.355; n = 750). Increased BMI, fasting glucose, insulin (at 120 min), total cholesterol (TC), and baPWV values were observed in Groups C2 and C3, with statistically significant trends (all trends P < 0.05). The CMI was positively correlated with systolic blood pressure (r = 0.74, P < 0.001). Multivariate analysis revealed that an increased CMI contributed to a greater risk for arteriosclerosis (OR = 1.87, 95%CI: 1.66-2.10, P < 0.001). Compared to the C1 group, the C2 group and C3 group had a greater risk of developing arteriosclerosis, with ORs of 4.55 (95%CI: 3.57-5.81, P<0.001) and 5.56 (95%CI: 4.32-7.17, P<0.001), respectively. The association was notably stronger in patients with a BMI below 21.62 kg/m² than in those with a BMI of 21.62 kg/m² or higher (OR = 4.53 vs. OR = 1.59). CONCLUSIONS: These findings suggest that the CMI is a relevant and independent marker of arteriosclerosis in patients with T2DM and may be useful in the risk stratification and management of these patients.

Effects of 60 days of 6° head-down bed rest on the composition and function of the human gut microbiota.

Spaceflight is rigorous and dangerous environment which can negatively affect astronauts' health and the entire mission. The 60 days of 6° head-down bed rest (HDBR) experiment provided us with an opportunity to trace the change of gut microbiota under simulated microgravity. The gut microbiota of volunteers was analyzed and characterized by 16S rRNA gene sequencing and metagenomic sequencing. Our results showed that the composition and function of the volunteers' gut microbiota were markedly was affected by 60 days of 6° HDBR. We further confirmed the species and diversity fluctuations. Resistance and virulence genes in the gut microbiota were also affected by 60 days of 6° HDBR, but the species attributions remained stable. The human gut microbiota affected by 60 days of 6° HDBR which was partially consistent with the effect of spaceflight, this implied that HDBR was a simulation of how spaceflight affects the human gut microbiota.

Preliminary investigation of hepatitis E virus detection by a recombinase polymerase amplification assay combined with a lateral flow strip.

Hepatitis E virus (HEV) is a zoonotic pathogen that is a significant public health problem. Detecting HEV relies mainly on conventional PCR, which is time-consuming and requires sophisticated instruments and trained staff. We aimed to establish a reverse-transcription (RT)-recombinase polymerase amplification (RPA) assay (RT-RPA) combined with a lateral flow strip (LFS; RT-RPA-LFS) to rapidly detect HEV RNA in human and rabbit samples. With the optimal reaction conditions (37°C for 30 min), our assay detected as few as 1.0 × 102 copies/mL of HEV and showed no cross-reactivity with other hepatitis viruses. We tested 28 human samples (4 fecal and 24 serum samples) and 360 rabbit samples (180 fecal and 180 serum samples) with our RT-RPA-LFS assay and compared our assay to an RT-qPCR method. There was no significant difference (p > 0.05) in the test results between the 2 assays. Our RT-RPA-LFS assay detected both HEV3 and HEV4 genotypes. Our rapid, sensitive, and specific RT-RPA-LFS assay for the detection of HEV may provide a useful detection tool for limited-resource areas.

Investigation of genotype diversity of 7,804 norovirus sequences in humans and animals of China.

Norovirus is a prominent enteric virus responsible for severe acute gastroenteritis disease burden worldwide. In our current study, we analyzed 7,804 norovirus sequences of human and animals in China which were detected from 1980 to 2020 from GenBank. The GenBank database was searched up to May 2021 with the following search terms: "norovirus" or "norwalk virus" and "China." The 7,804 norovirus sequences were collected and evaluated by phylogenetic analysis using MEGA X software package. The online typing tool (https://www.rivm.nl/mpf/typingtool/norovirus/) was used to confirm the genotypes. There were 36 norovirus genotypes prevailing in China. GII.4 was the most prevalent genotype, and GII.2, GII.3 and GII.17 also emerged during different time periods. Most sequences were detected in East China (41.72%, 3,256/7,804), but different norovirus genotypes were distributed widely across the country. A variety of norovirus genotypes, including GI, GII, GIII, GIV, GV, GVI, GVII and GX, were reported in different animals. Furthermore, a GI.3 sequence detected from animal had high identity with norovirus detected in human from the same region, indicating the potential norovirus zoonotic transmission in China. In conclusion, these results indicated that norovirus sequences with considerable genetic diversity distributed widely in China, with potential reverse zoonotic transmission from human to animals.

Characterization of highly expressed novel hub genes in hepatitis E virus chronicity in rabbits: a bioinformatics and experimental analysis.

BACKGROUND: Hepatitis E virus (HEV), which is the leading cause of acute viral hepatitis worldwide, usually causes self-limited infections in common individuals. However, it can lead to chronic infection in immunocompromised individuals and its mechanisms remain unclear. Rabbits are the natural host of HEV, and chronic HEV infections have been observed in rabbits. Therefore, we aimed to investigate potential key genes in HEV chronicity process in rabbits. In this study, both bioinformatics and experimental analysis were performed to deepen the understanding of hub genes in HEV chronic infection in rabbits. RESULTS: Ninety-four candidate differentially expressed genes (DEGs) and the pathways they enriched were identified to be related with HEV chronicity. A total of 10 hub genes were found by protein-protein interaction (PPI) network construction. Rabbits of group P (n = 4) which showed symptoms of chronic HEV infection were selected to be compared with HEV negative rabbits (group N, n = 6). By detecting the identified hub genes in groups P and N by real-time PCR, we found that the expressions of MX1, OAS2 and IFI44 were significantly higher in group P (P < 0.05). CONCLUSIONS: In this work, we presented that MX1, OAS2 and IFI44 were significantly upregulated in HEV chronic infected rabbits, indicating that they may be involved in the pathogenesis of HEV chronicity.

Prognostic Factors and Clinical Characteristics of Chronic Hepatitis B with or without Nucleos(t)ide Analogues Therapy: A Retrospective Study.

OBJECTIVE: We compared the prognostic factors and clinical characteristics of chronic hepatitis B (CHB) patients who received nucleos(t)ide analogues (NAs) therapy to those who did not. METHOD: A total of 315 CHB patients were enrolled in this study and were divided into NA (n=144) and non-NA (n=171) groups based on their therapy. RESULTS: The risk of hepatocellular carcinoma (HCC) development and mortality in the NA group were significantly lower than those in the non-NA group. Smoking, alcohol abuse, AFP, γ-GTP, and HBV DNA levels were significantly correlated with hepatocarcinogenesis. Alcohol abuse, AFP, γ-GTP, HBV DNA levels and NA treatment were associated with mortality in these patients. CONCLUSIONS: NA therapy reduced the risk of HCC and mortality in CHB patients. Smoking, alcohol abuse, AFP >20 ng/mL, HBV DNA >20,000 IU/mL, and elevated γ-GTP serum concentration were significantly related to poor outcomes.

Fecal Bile Acids Profile of Crewmembers Consuming the Same Space Food in a Spacecraft Simulator.

Introduction: Recently, bile acids (BAs) are increasingly being considered as unique metabolic integrators and not just for the cholesterol metabolism and absorption of dietary lipids. Human BAs profiles are evolved to be individual under different environmental, dietary, and inherited factors. Variation of BAs for crewmembers from freshly prepared kitchen diets to wholly prepackaged industrial foods in a ground-based spacecraft simulator has not been clearly interpreted. Methods: Three crewmembers were confined in a docked spacecraft and supplied with 7 days periodic wholly prepackaged industrial foods for 50 days. Fecal samples were collected before entry in the spacecraft simulator and after evacuation. Determination of 16 kinds of BAs was carried out by high-performance liquid chromatography tandem mass spectrometry method. Results: Bile acids metabolism is sensitive to diet and environment transition from freshly prepared kitchen diets in the canteen to wholly prepackaged industrial foods in a ground-based spacecraft simulator, which is also specific to individuals. A significant positive relationship with a coefficient of 0.85 was found for primary BAs as chenodeoxycholic acid (CDCA) and cholic acid (CA), and a significantly negative relationship with a coefficient of -0.69 for secondary BAs as lithocholic acid (LCA) and deoxycholic acid (DCA). Discussion: The profile of BA metabolism of individuals who share the same food in the same environment appears to be unique, suggesting that the inherent ability of different individuals to adapt to diet and environment varies. Since the transition from the free diet in open space to whole prepackaged space food diet in a space station simulator causes the variations of BAs pool in an individual manner, assessment of BA metabolic profiles provides a new perspective for personalized diet design, astronaut selection and training, and space flight diet acclimatization.

Case Report: A Novel ABCC8 Variant in a Chinese Pedigree of Maturity-Onset Diabetes of the Young.

Background: We aimed to analyze a novel ABCC8 variant of a Chinese patient with suspected maturity-onset diabetes of the young (MODY) and to provide evidence for precise diagnosis and appropriate treatment. Method: A Chinese family with suspected MODY was recruited in this study, which included a 15-year-old female patient with diabetes. Clinical data and blood samples were collected from the proband and other family members. All of the living relatives were given an oral glucose tolerance test. Next-generation sequencing was performed to identify the mutated genes in the proband. Sanger sequencing was utilized to confirm the location of the pathogenic variant in all subjects. Further treatment was referred to targeted family members according to genetic testing. Results: The proband was found to have a random blood glucose level of 244.8 mg/dl and an HbA1c level of 9.2%. Before this investigation, her grandparents had been diagnosed with diabetes. The second uncle, two aunts, mother, and cousin of the proband were diagnosed with diabetes by abnormal HbA1C (6.5-12.1%) and fasting blood glucose (FBG, 91.4-189.7 mg/dl). The second aunt of the proband had impaired glucose homeostasis (HbA1C = 6.4% and FBG = 88.0 mg/dl). One novel missense variant c.1432G>A (p.A478T) in exon 9 of the ABCC8 gene was detected in the proband with suspected MODY. The variant was also found in six family members with diabetes or impaired glucose homeostasis, including her second uncle, two aunts, mother, and cousin. After the treatment was switched to glimepiride, the fasting blood glucose was adjusted to 99.54 mg/dl, the 2-h postprandial blood glucose was 153.54 mg/dl, serum fructosamine was 259 µmol/l, and HbA1c was 5.8%. The glycemic control remained optimal, and no hypoglycemic episodes were observed in the living relatives. Conclusion: This study revealed one novel missense variant of the ABCC8 gene in Chinese families. The present findings indicated that the members of this family responded to treatment with sulfonylureas as previously seen in ABCC8 MODY.

Effects of spaceflight on the composition and function of the human gut microbiota.

Interaction between humans and the gut microbiota is important for human physiology. Here, the gut microbiota was analyzed via metagenomic sequencing, and the fluctuations in the gut microbiota under the conditions of spaceflight were characterized. The composition and function of the gut microbiota were substantially affected by spaceflight; however, individual specificity was uncompromised. We further confirmed the species fluctuations and functional genes from both missions. Resistance and virulence genes in the gut microbiota were affected by spaceflight, but the species attributions remained stable. Spaceflight markedly affected the composition and function of the human gut microbiota, implying that the human gut microbiota is sensitive to spaceflight.

Identification of peptides that bind hepatitis C virus envelope protein E2 and inhibit viral cellular entry from a phage-display peptide library.

Hepatitis C virus (HCV) envelope protein E2 is required for the entry of HCV into cells. Viral envelope proteins interact with cell receptors in a multistep process, which may be a promising target for the development of novel antiviral agents. In this study, a heptapeptide M13 phage-display library was screened for peptides that bind specifically to prokaryotically expressed, purified truncated HCV envelope protein E2. ELISA assay was used to quantify the binding of the peptides to HCV E2 protein. Flow cytometry, quantitative reverse-transcription PCR and western blotting were used to investigate the inhibition effect of one peptide on HCV infection in hepatoma cells (Huh7.5) in vitro. Four peptides capable of binding specifically to HCV E2 protein were obtained after three rounds of biopanning. Peptide C18 (WPWHNHR), with the highest affinity for binding HCV E2 protein, was synthesized. The results showed that peptide C18 inhibited the viral infectivity of both HCV pseudotype particles (HCVpp) harboring HCV envelope glycoproteins and cell-culture produced HCV (HCVcc). Thus, this study demonstrated that peptide C18 is a potential candidate for anti-HCV therapy as a novel viral entry inhibitor.

Cellular immunogenicity of a multi-epitope peptide vaccine candidate based on hepatitis C virus NS5A, NS4B and core proteins in HHD-2 mice.

To develop a vaccine against hepatitis C virus (HCV), a multi-epitope peptide was synthesized from nonstructural proteins containing HLA-A2 epitopes inducing mainly responses in natural infection. The engineered vaccine candidate, VAL-44, consists of multiple epitopes from the HCV NS5A, NS4B and core proteins. Immunization with the VAL-44 peptide induced higher CTL responses than those by the smaller VL-20 peptide. VAL-44 induced antigen-specific IFN-γ-producing CD4+ T cells and CD8+ T cells. VAL-44 elicited a Th1-biased immune response with secretion of high amounts of IFN-γ and IL-2, compared with VL-20. These results suggest that VAL-44 can elicit strong cellular immune responses. The VAL-44 peptide stimulated IFN-γ production from viral-specific peripheral blood mononuclear cells (PBMCs) of patients infected with HCV. These results suggest that VAL-44 could be developed as a potential HCV multi-epitope peptide vaccine.

[Immune response in BALB/c mice immunized with BCG expressing HBV truncated C gene and preS1 gene].

AIM: To study expressing of HBV truncated core and preS1 protein in BCG and to explore the effect of humoral and cellular immune response stimulated by the recombinant BCG. METHODS: A shuttle vector was constructed and was transformed into BCG which including truncated Core gene and preS1 gene of HBV. The recombinant BCGs were analyzed with SDS-PAGE and Western blot. The three groups of BALB/c mice were immunized with saline, BCG, BCG-pDE22 and BCG-pDE22-CS1 respectively. Then the antibody titer and CTL effects were evaluated. RESULTS: Compared with the control group, the recombinant BCG could express a new protein band of 24 kD which was consistent with the size of CS1 fused protein which displayed a good antigen-binding property by Western blot analysis. The antibody titer significantly increased and CTL effect apparently enhanced in the recombinant BCG immunized group. CONCLUSION: BCG could be as live vector which carrying HBV related genes, which developing a novel vaccine against HBV.

[Evaluation on the analytical sensitivity of 31 HBsAg enzyme immunoassay kits].

OBJECTIVE: To study the analytical sensitivity on 31 HBsAg enzyme immunoassy (EIA) test kits. METHODS: Thirty one HBsAg EIA kits produced by domestic or overseas manufactories and applied for approval during May 2007 to May 2008, were evaluated using the national reference panels. The hyperbolic curve of the log A value and log concentration for the national sensitivity standards was established. The cut-off value of each kit was substituted into the curvilinear equation to determine the analytical sensitivity which was compared between different HBsAg EIA kits. RESULTS: Twenty seven (351 lots) domestic and 4 (27 lots) overseas kits were compared. Among 378 lots of the 31 HBsAg EIA kits, only 2 lots of the domestic kits had a lower sensitivity when tested with the national HBsAg reference panels, with an average approvalr ate of 99.43% (349/351). The mean analytical sensitivity of the domestic kits for adr, adw, ay serotypes were 0.307, 0.419, 0.513 ng/ml, respectively. There was a significant difference between serotypes (F = 97.30, P < 0.01). The mean analytical sensitivity of the overseas kits for adr, adw, ay serotypes were 0.054, 0.066, 0.050 ng/ml respectively, with no significant difference between serotypes (F = 0.65, P > 0.05). The analytical sensitivity of the overseas kits for all the three serotypes was higher than that of the domestic kits (P < 0.01). There was no significant difference found between the analytical sensitivities of the kits produced by the same manufactory using 30- or 60-minute incubation of detection (P > 0.05). In contrast, there was significant difference noticed between the analytical sensitivities of the kits produced by the same manufactory when tested for 10 or 15-minute coloration of the results (P < 0.01). CONCLUSION: Analytical sensitivity of the HBsAg EIA domestic kits should be further improved, especially for detecting adw and ay serotypes.

[Primary evaluation of anti-HEV diagnostic reagent by experimental infection animal model with hepatitis E virus].

OBJECTIVE: To evaluate anti-HEV diagnostic kits by experimental infecting rhesus monkeys with HEV. METHODS: Eight rhesus monkeys were infected with genotype 1 and 4 HEV separately. The alanine aminotransferase (ALT) level of all monkeys were detected before and after the process of infection. HEV RNA in stool specimens was tested by reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Anti-HEV IgG in serum was detected by GL-IgG and WT-IgG. RESULTS: HEV RNA presented in the stool of all the 8 monkeys after infection. The ALT level of 1 monkey infected with genotype 1 HEV and 2 monkeys infected with genotype 4 HEV appeared abnormally after infection. Tested by GL-IgG, 2 of the 4 monkeys infected with genotype 1 HEV and 1 of 4 monkeys infected with genotype 4 HEV seroconverted to anti-HEV IgG. However, when tested by WT-IgG, all the infected monkeys seroconverted to anti-HEV IgG. The anti-HEV IgG tested by WT-IgG was positive during the whole observation period,and the anti-HEV IgG measured by GL-IgG only remained 12 weeks after infection. Detected by GL-IgG and WT-IgG, seropositive conversion of the anti-HEV IgG happened almost at the same time. CONCLUSION: Both GL-IgG and WT-IgG could detect the anti-HEV IgG of experimentally infected rhesus monkeys but the WT-IgG had a higher sensitivity for detection of anti-HEV IgG than

[Evaluation of multiplex nucleic acid testing assays for screening of hepatitis B virus DNA in blood donation process].

OBJECTIVE: To evaluate the multiplex nucleic acid testing (NAT) assays for HBV, HCV and HIV in detecting HBV DNA in plasma samples. METHODS: 534 plasma samples collected form several areas were detected with Abbott Architect i2000 HBsAg, ani-HBs, HBeAg, anti-HBe, anti-HBc and anti-HBc IgM diagnostic kits. HBV DNA levels of those samples were detected with Roche COBAS AmpliPrep/COBAS TaqMan HBV Test. Two kinds of multiplex NAT assays for HBV, HCV and HIV were used to test HBV DNA of those 534 samples. Results of serology-markers and quantitative HBV DNA levels with results of NAT were compared. RESULTS: HBV DNA was positive in all 81 HBsAg, HBeAg and anti-HBc positive samples, detected by both of NAT assays. HBV DNA was positive in 11 and 19 of 200 HBsAg negative samples when detected with the two kinds of NAT assays separately. Compared with the quantitative results detected by Roche COBAS AmpliPrep/COBAS TaqMan HBV Test, the HBV DNA positive rates were 96.9% and 94.3% in 193 samples of HBV DNA levels over 500 IU/ml while 40.2% and 45.3% in 117 samples of HBV DNA levels below 500 IU/ml while 99.3% and 96.0% in 151 samples of DNA negative HBV. CONCLUSION: There are some occult low level HBV DNA carriers with HBsAg negative results in China. NAT assays for HBV, HCV and HIV may be useful to improve the transfusion safety.

[Clinical significance of detecting RNA and anti HEV antibody in convalesent sera in patients with acute HEV hepatitis].

OBJECTIVE: To investigate the anti hepatitis E virus (HEV) and HEV RNA in acute and convalescent sera of patients with NonA-E acute hepatitis. METHODS: The serum samples were taken from 95 patients who were diagnosed as acute NonA-E hepatitis. Enzyme immunoassay (EIA) was used for detecting anti-HEV Immunoglobulin G (IgG, Genolable and Wantai EIA anti-HEV kits). RT-PCR amplification of HEV RNA was based on the open reading frame 2 region of HEV and the PCR products were sequenced. RESULTS: Sera from 95 patients who were negative for anti-HEV in acute phase were followed up for 11-35 days to detect the anti-HEV antibody in recovery phase, 16/95 (16.84%) were positive for anti-HEV (wantai EIA anti-HEV kits). Ten (62.50%) were positive for HEV RNA in acute phase. Sequence analysis showed that 4 were HEV genotype. 6 were HEV genotype; 12/95 (12.50%) were positive for anti-HEV (Genolable EIA anti-HEV kits). Seven were positive for HEV RNA; 4 belonged to HEV genotype, 3 were HEV genotype. CONCLUSION: It is significant and necessary to detect anti HEV antibody and HEV RNA in patients with HEV infection during acute phase and convalesent phase.

[Preparation and application of the national reference panel for hepatitis B virus DNA].

OBJECTIVES: To calibrate the national hepatitis B virus (HBV) DNA standard according to world health organization's standard material and prepare the national reference panel for HBV DNA reagents. METHODS: Sera from blood donors and HBV patients were collected and detected by home-made HBV DNA PCR kits, HBsAg kits, and anti-HBc kits, and then confirmed by HBV DNA PCR kits produced by Roche in German, which was recognized by the world health organization. The stability of the panel was detected by acceleration method. RESULTS: The convinced copies of the sensitivity samples were gotten by seven independent experiments, the coefficients of variation of logarithm of the copies of L0-L5 were all less than 15%. Regarding the national reference panel as the standard, the quality of most domestic HBV DNA PCR kits was improved, while part of the kits should be further qualified. CONCLUSION: The national reference panel for HBV DNA reagents is developed. It contains eight negative, nine positive sera and seven samples for sensitivity test

Prevalence, isolation, and partial sequence analysis of hepatitis E virus from domestic animals in China.

Evidence that hepatitis E is zoonotic is accumulating. Serum samples were collected from pigs, cattle, and goats from various regions of China to determine whether they had been infected with hepatitis E virus (HEV). An in-house enzyme immunoassay (EIA) and reverse transcriptase-polymerase chain reaction (RT-PCR) with primers from open reading frame (ORF) 2 were used to detect anti-HEV antibodies and HEV RNA. The mean positivity rates of anti-HEV antibody for pigs and cattle were 78.8% and 6.3% but none of the goat sera were positive. Pigs may be more susceptible to infection with HEV than cattle or goats. Five of 263 pig sera were positive for HEV RNA and four of these five were also positive for anti-HEV. The PCR products (nt 6007-6354) were cloned and sequenced and compared to other HEV sequences in the nucleotide databases. The five sequences shared 83-93% identity to each other at the nucleotide level and 74-79%, 73-74%, 73-78%, and 83-99% identity to HEV genotypes 1, 2, 3, and 4, respectively. They were closely related to human isolates of HEV genotype 4. Phylogenetic analyses also place these swine sequences in HEV genotype 4, resembling most closely viruses isolated from Chinese patients with acute hepatitis. These data support the hypothesis that sporadic hepatitis E in China is zoonotic.