Survey of physicians' knowledge about pediatric nonalcoholic fatty liver disease in China.

OBJECTIVES: To evaluate physicians' awareness and knowledge towards pediatric nonalcoholic fatty liver disease (NAFLD) and their attitude toward change in nomenclature from NAFLD to metabolic dysfunction-associated fatty liver disease (MAFLD) or metabolic dysfunction-associated steatotic liver disease (MASLD) in China. METHODS: The questionnaire survey contained five parts (characteristics of the participants, epidemiology, diagnosis, management of NAFLD, and attitudes toward the nomenclature of MAFLD/MASLD). The participants included 53 hepatologists, 88 gastroenterologists (GEs), 74 endocrinologists (ENDOs), 61 primary care physicians (PCPs), and 157 pediatricians across 31 municipalities, provinces and autonomous regions of China's mainland. RESULTS: Hepatologists saw the largest number of pediatric NAFLD patients annually (median 9 [range 1-20]), with the lowest number by PCPs (even notwithstanding one patient annually). The primary sources of pediatric NAFLD knowledge were acquired via guidelines. Hepatologists had the highest total knowledge score among all five types of physicians. Approximately one-third of nonspecialists (ENDOs and PCPs) considered liver biopsy necessary for pediatric NAFLD patients, and this percentage increased to half in specialists (hepatologists and GEs). For nonspecialists, the major barriers to the management of pediatric NAFLD were poor patient adherence to lifestyle modifications and lacking confidence in managing NAFLD. Above 90% physicians agreed to change the nomenclature NAFLD to MAFLD; however, they were not sure whether it could reduce the economic burden. CONCLUSIONS: Despite the epidemic of pediatric NAFLD in China, a significant knowledge gap remains in the identification, diagnosis, and treatment of pediatric NAFLD, particularly among frontline workers such as pediatricians and PCPs. More education programs should be carried out in the future.

Artificial Intelligence Model for Antiinterference Cataract Automatic Diagnosis: A Diagnostic Accuracy Study.

Background: Cataract is the leading cause of blindness worldwide. In order to achieve large-scale cataract screening and remarkable performance, several studies have applied artificial intelligence (AI) to cataract detection based on fundus images. However, the fundus images they used are original from normal optical circumstances, which is less impractical due to the existence of poor-quality fundus images for inappropriate optical conditions in actual scenarios. Furthermore, these poor-quality images are easily mistaken as cataracts because both show fuzzy imaging characteristics, which may decline the performance of cataract detection. Therefore, we aimed to develop and validate an antiinterference AI model for rapid and efficient diagnosis based on fundus images. Materials and Methods: The datasets (including both cataract and noncataract labels) were derived from the Chinese PLA general hospital. The antiinterference AI model consisted of two AI submodules, a quality recognition model for cataract labeling and a convolutional neural networks-based model for cataract classification. The quality recognition model was performed to distinguish poor-quality images from normal-quality images and further generate the pseudo labels related to image quality for noncataract. Through this, the original binary-class label (cataract and noncataract) was adjusted to three categories (cataract, noncataract with normal-quality images, and noncataract with poor-quality images), which could be used to guide the model to distinguish cataract from suspected cataract fundus images. In the cataract classification stage, the convolutional-neural-network-based model was proposed to classify cataracts based on the label of the previous stage. The performance of the model was internally validated and externally tested in real-world settings, and the evaluation indicators included area under the receiver operating curve (AUC), accuracy (ACC), sensitivity (SEN), and specificity (SPE). Results: In the internal and external validation, the antiinterference AI model showed robust performance in cataract diagnosis (three classifications with AUCs >91%, ACCs >84%, SENs >71%, and SPEs >89%). Compared with the model that was trained on the binary-class label, the antiinterference cataract model improved its performance by 10%. Conclusion: We proposed an efficient antiinterference AI model for cataract diagnosis, which could achieve accurate cataract screening even with the interference of poor-quality images and help the government formulate a more accurate aid policy.

Effect of the bacterial community on the volatile flavour profile of a Chinese fermented condiment - Red sour soup - During fermentation.

Red sour soup (RSS) is a traditional fermented seasoning used by people in Guizhou Province, China. The volatile compounds were detected by gas chromatography ion mobility spectrometry (GC-IMS), while the bacterial community compositions were revealed by 16S sequencing. A total of 70 volatile substances were assessed, and esters, terpenes, and alcohols played a dominant role in RSS. Lactic acid bacteria (LAB) were dominant in the microbial community. During fermentation, multiple volatile flavour substances and various LAB were mainly derived from the secondary fermentation stage. The core bacteria with an important influence on volatile flavour substances were analysed by the Mantel test, identifying Bacteroides, Lactobacillus, Lactococcus, Enterobacteriaceae, Pantoea, Clostridium, Enterobacter, unclassified genera, Caproiciproducens, Nitriliruptoraceae, Halomonas, Bradyrhizobium, Pediococcus, Caulobacteraceae, Weissella, Ligilactobacillus, and Levilactobacillus and et al.. This study provides us with useful information about flavour-generating bacteria among RSS fermentation periods. The abundances of these bacteria may be controlled to enrich the desired flavour metabolites while eliminating unwanted metabolites.

Dynamic SARS-CoV-2-Specific Immunity in Critically Ill Patients With Hypertension.

Background: The current outbreak of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses an unprecedented health crisis. The most common chronic illness among patients infected with SARS-CoV-2 is hypertension. Immune dysregulation plays an important role in SARS-CoV-2 infection and in the development of hypertension; however, the dynamic immunological characteristics of COVID-19 patients with hypertension remain largely unclear. Methods: In total, 258 hypertensive patients infected with SARS-CoV-2 were included in this study. CD38+HLA-DR+ and CD38+PD-1+ CD8+ T cells, IFNγ+CD4+ and IFNγ+CD8+ T cells, the titers of IgG, IgM, and IgA against SARS-CoV-2 spike protein, and SARS-CoV-2 throat viral loads were measured weekly over 4 weeks after the onset of symptoms. Clinical outcomes were also monitored. Findings: CD4+ T lymphopenia was observed in 100% of the severe and critical cases. Compared with the surviving patients, the patients with fatal outcomes exhibited high and prolonged expression of CD38+HLA-DR+ and CD38+PD-1+ on CD8+ T cells, low expression of SARS-CoV-2-specific IFNγ+CD4+ and IFNγ+CD8+ T cells, low titers of IgG, IgM, and IgA against SARS-CoV-2 spike protein, and high SARS-CoV-2 viral load during the illness. In the surviving patients, the viral load was significantly inversely correlated with SARS-CoV-2-specific IFNγ+CD8+and IFNγ+CD4+ T cells, IgG, IgM, and IgA. Interpretation: T lymphopenia is common in critical or severe COVID-19 cases with hypertension. Prolonged activation and exhaustion of CD8+ T cells were associated with severe disease. The delayed SARS-CoV-2-specific antibody responses may be insufficient for overcoming severe SARS-CoV-2 infection in the absence of SARS-CoV-2-specific cellular responses.

Deep insights into fungal diversity in traditional Chinese sour soup by Illumina MiSeq sequencing.

Sour soup is a traditional condiment in Guizhou Province, China. The purpose of this study was to investigate the differences in the fungi present in 5 types of sour soup (tomato sour soup, chili sour soup, cherry tomato sour soup, spoiled tomato sour soup, and red sour soup made from blended tomato and chili sour soup subjected to secondary fermentation) and to determine the reasons for the deterioration of tomato sour soup by comparing the fungal communities in normal and deterioratedtomato sour soup. A total of 5 phyla were detected in all 5 samples, including Ascomycota (69.38%), Basidiomycota (7.63%), Zygomycota (1.59%), Chytridiomycota (0.01%) and unclassified phyla (21.39%). Ascomycota was the main phylum in each sample except the red sour soup made from blended tomato and chili sour soup subjected to secondary fermentation. That sour soup contained many unrecognized phyla. At the genus level, there were major differences among the different samples. Dekkera spp. and Pichia spp. were the main dominant fungus in tomato sour soup, Saccharomyces spp. and Pichia spp. were the dominant fungus in chili sour soup, and Pichia spp. were the dominant fungus in cherry tomato sour soup. When sour soup went bad, the fungus of sour soup changed greatly, and the unknown fungal genera, Cladospora spp., Saccharomyces spp. and Emericella spp. became the dominant fungal genera. In addition, after the secondary fermentation of tomato and chili sour soup mixed with garlic and ginger, the fungal genera of the base fermentation were replaced by unknown fungal genera. Moreover, there were various spoilage fungi in sour soup, which indicated that there were safety risks in naturally fermented sour soup and should be further controlled. This study revealed the fungal flora in sour soup made from different vegetables and compared the fungal diversity of spoiled and normal tomato sour soup and thereby provided a basis for understanding the fungal diversity of sour soup in China and guiding the production of sour soup.

Deep Insights into Gut Microbiota in Four Carnivorous Coral Reef Fishes from the South China Sea.

Investigations of gut microbial diversity among fish to provide baseline data for wild marine fish, especially the carnivorous coral reef fishes of the South China Sea, are lacking. The present study investigated the gut microbiota of four carnivorous coral reef fishes, including Oxycheilinus unifasciatus, Cephalopholis urodeta, Lutjanus kasmira, and Gnathodentex aurolineatus, from the South China Sea for the first time using high-throughput Illumina sequencing. Proteobacteria, Firmicutes, and Bacteroidetes constituted 98% of the gut microbiota of the four fishes, and 20 of the gut microbial genera recovered in this study represent new reports from marine fishes. Comparative analysis indicated that the four fishes shared a similar microbial community, suggesting that diet type (carnivorous) might play a more important role in shaping the gut microbiota of coral reef fishes than the species of fish. Furthermore, the genera Psychrobacter, Escherichia-Shigella, and Vibrio constituted the core microbial community of the four fishes, accounting for 61-91% of the total sequences in each fish. The lack of the genus Epulopiscium in the four fishes was in sharp contrast to what has been found in coral reef fishes from the Red Sea, in which Epulopiscium was shown to be the most dominant gut microbial genus in seven herbivorous coral reef fishes. In addition, while unique gut microbial genera accounted for a small proportion (8-13%) of the total sequences, many such genera were distributed in each coral reef fish species, including several genera (Endozoicomonas, Clostridium, and Staphylococcus) that are frequently found in marine fishes and 11 new reports of gut microbes in marine fishes. The present study expands our knowledge of the diversity and specificity of gut microbes associated with coral reef fishes.

A novel HLA class I allele, HLA-B*13:02:21, identified by sequence-based typing in a Chinese individual.

HLA-B*13:02:21 has one nucleotide substitution, C > T(GAC to GAT) compared with B*13:02:01.

The unexpected diversity of microbial communities associated with black corals revealed by high-throughput Illumina sequencing.

The microbes associated with black corals remain poorly studied. The present study is the first attempt to investigate microbial community structure in the black corals Antipathes ceylonensis and A. dichotoma from the South China Sea by using high-throughput Illumina sequencing. A total of 52 bacterial and 3 archaeal phyla were recovered in this study, suggesting the black corals harboured highly diverse microbial communities. Among the 55 microbial phyla, Proteobacteria, Firmicutes, Bacteroidetes, Chloroflexi, Acidobacteria and Actinobacteria dominated in the two black corals from the South China Sea. Although most of the microbial phyla recovered from the two black corals have been reported in previous studies on coral-associated microbes, eight bacterial phyla including Synergistetes, Thermi, AncK6, GNO2, NKB19, NC10, WWE1 and GAL15, and the archaeal phylum Parvarchaeota are reported for the first time from corals in this study, which expands our knowledge about the diversity of coral-associated microbes. The comparison of microbial communities in the different black coral species indicated that A. ceylonensis harboured few abundant bacterial genera such as Citrobacter and Pseudomonas, whereas a high diversity of rare bacterial genera (<1% abundance), such as Winogradskyella and Rubricoccus, was detected only in A. dichotoma. These results suggested that the microbial community in black corals exhibited species-specific variation.

Modulation of human melanoma cell proliferation and apoptosis by hydatid cyst fluid of Echinococcus granulosus.

OBJECTIVE: The objective of this paper was to assess the effects of hydatid cyst fluid (HCF) of Echinococcus granulosus on melanoma A375 cell proliferation and apoptosis. METHODS: A375 cells were classified into five groups by in vitro culture: normal group, control group, 10% HCF group, 20% HCF group and 30% HCF group. Trypan blue staining method was employed to detect the toxicity of HCF. Effects of different concentrations of HCF on melanoma A375 cell proliferation at different time points were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Flow cytometry and propidium iodide (PI) staining were used to detect cell cycle, and Annexin-V/PI double staining method was used to determine A375 cell apoptotic rate. Western blotting was applied to detect the expression of phosphorylated extracellular regulated protein kinases, proliferating cell nuclear antigen (PCNA), cell-cycle-related proteins (cyclin A, cyclin B1, cyclin D1 and cyclin E) and apoptosis-related proteins (Bcl-2, Bax and caspase-3). RESULTS: HCF with a high concentration was considered as atoxic to A375 cells. HCF promoted A375 cell proliferation, and the effects got stronger with an increase in concentrations but was retarded after reaching a certain range of concentrations. HCF increased phosphorylation level and expression of extracellular regulated protein kinase, as well as PCNA expression. HCF also promoted the transferring progression of A375 cells from the G0/G1 phase to the S phase to increase the cell number in S phase and increased the expression of cyclin A, cyclin D1 and cyclin E. HCF increased the expression of procaspase-3 (the precursor of apoptosis-related protein caspase-3) and antiapoptotic protein-Bcl-2, and decreased the expression of proapoptotic factor Bax, thereby inhibiting cell apoptosis. CONCLUSION: As a result, this study confirmed that HCF promotes proliferation and inhibits apoptosis of melanoma A375 cells.

Knockdown of MPP8 suppresses cell proliferation via regulation of HOXA5 in non-small cell lung cancer cells.

M-phase phosphoprotein 8 (MPP8) is reported to be closely implicated in cancer initiation and progression. In addition, the homeobox gene HOXA5 has been shown to play critical roles in hematopoiesis, embryogenesis, and tumorigenesis. Nevertheless, the functional relevance of MPP8 and it's relation with HOXA5 in non-small cell lung cancer (NSCLC) is unknown. Therefore, the present study aimed to detect the expression profile of MPP8 in NSCLC and further explore it's biological roles in lung cancer cells. Cell proliferation was measured by CCK-8 assay and EdU incorporation assay. Real-time PCR was applied to detect the mRNA expression of MPP8 and HOXA5. The protein levels of MPP8 and HOXA5 were evaluated by western blot. Our study found that the expression of MPP8 was significantly increased in the NSCLC tissue compared with the adjacent non-tumorous tissue. Compared with the human lung fibroblasts, the elevated gene expression of MPP8 was also detected in the human NSCLC cell lines including NCI-H23 and NCI-H1299. In addition, knockdown of MPP8 led to an obvious reduction in cell viability and DNA synthesis in NCI-H23 and NCI-H1299 cells. Furthermore, down-regulation of MPP8 resulted in elevated expression of HOXA5 in NSCLC cells both at the mRNA and protein levels. Moreover, depletion of HOXA5 abolished the anti-tumor function of MPP8 knockdown in NSCLC cells. The present study demonstrated that MPP8 was associated with NSCLC cell proliferation through regulation of HOXA5, suggesting that MPP8 may act as a novel therapeutic target for treatment of NSCLC.

Effect of combined treatment with cyclophosphamidum and allicin on neuroblastoma-bearing mice.

OBJECTIVE: To evaluate the efficacy of allicin combined with cyclophosphamide on neuroblastoma (NB)-bearing mice and explore the immunological mechanism in it. METHODS: A total of 30 NB-bearing mice were equally randomized into model group, cyclophosphamide group and combined therapy group, 10 nudemice were set as normal saline (NS) group. Cyclophosphamide group and combined therapy group were weekly injected with 60 mg/kg cyclophosphamide for four weeks; besides, combined therapy group was given with allicin (10 mg/kg/d) by gastric perfusion for 4 weeks; model group and NS group were given with the same volume of NS. Serum VEGF content was detected by ELISA pre-treating (0 d) and on the 3rd d, 14th d and 28th d; on 29th d, all mice were sacrificed and the tumor, liver, spleen and thymic tissues were weighted. Tumors were made into paraffin section for detecting tumor cell apoptosis and proliferation by TUNEL and BrdU method, respectively. Survival curves were drawn by Kaplan-Meier method. RESULTS: After treatment, both treatment groups relieved on viscera indexes, VEGF level, T cell subsets distribution and tumor growth and each index of combined therapy group was better than cyclophosphamide group (P<0.05 or 0.01); only combined therapy group could significantly increase the lifetime of NB-bearing mice (µ (2)=5.667, P=0.017). CONCLUSIONS: Allicin can improve T cell subsets distribution and inhibit VEGF expression through its immunomodulatory activity, thereby improve the efficiency on NB in coordination with cyclophosphamide.

Statistical optimization of alkaline protease production from Penicillium citrinum YL-1 under solid-state fermentation.

Proteases from halotolerant and halophilic microorganisms were found in traditional Chinese fish sauce. In this study, 30 fungi were isolated from fermented fish sauce in five growth media based on their morphology. However, only one strain, YL-1, which was identified as Penicillium citrinum by internal transcribed spacer (ITS) sequence analysis, can produce alkaline protease. This study is the first to report that a protease-producing fungus strain was isolated and identified in traditional Chinese fish sauce. Furthermore, the culture conditions of alkaline protease production by P. citrinum YL-1 in solid-state fermentation were optimized by response surface methodology. First, three variables including peptone, initial pH, and moisture content were selected by Plackett-Burman design as the significant variables for alkaline protease production. The Box-Behnken design was then adopted to further investigate the interaction effects between the three variables on alkaline protease production and determine the optimal values of the variables. The maximal production (94.30 U/mL) of alkaline protease by P. citrinum YL-1 took place under the optimal conditions of peptone, initial pH, and moisture content (v/w) of 35.5 g/L, 7.73, and 136%, respectively.

Overcoming paclitaxel resistance in lung cancer cells via dual inhibition of stathmin and Bcl-2.

Lung cancer is the leading cause of death from malignancy in people and over 85% of these patients eventually die from disseminated disease. Paclitaxel (TAX) is widely used as an antimicrotubule agent for the treatment of lung cancer. Unfortunately, the resistance to this antimicrotubule agent occurs frequently. Stathmin (STMN1) is a ubiquitous microtubule destabilizing protein linked to cancer and cell health and its expression level often correlates with cancer stage progression and prognosis for survival. Overexpression of the antiapoptotic protein Bcl-2 has been shown to prolong drug-induced growth arrest, potentially inducing resistance. In this study, we used a short hairpin RNA (shRNA) approach to evaluate the effect of STMN1 and Bcl-2 downregulation in the sensitivity to TAX in lung cancer cells. We achieved significant downregulation of STMN1 and Bcl-2 mRNA and protein expression by a combination of double shRNA treatment strategy. Our experimental data showed that inhibition of STMN1 and Bcl-2 expression with RNA interference can sensitize lung cancer cells to TAX. These findings suggest a novel approach to improve the efficacy of certain antimicrotubule agents against lung cancer by regulating the function of STMN1 and Bcl-2.

[Appraisal of the repair gastroschisis with autogenous umbilical cord].

OBJECTIVE: To review the treatment of repair gastroschisis with autogenous umbilical cord in 13-year and evaluate its effect. METHODS: Twenty-two newborns who underwent the repair gastroschisis with autogenous umbilical cord between 1992 and 2005. The physical growth, intelligence measuring, area of operation in abdomen in the survived 18 cases were observed and followed-up. RESULTS: Eighteen patients recovered uneventfully, survival rate is 82%, their growth is well. They all developed incisional hernia near the operation, 9 cases recovered himself, 2 cases was operated to repair the abdominal hernia, 7 cases is under observed. CONCLUSIONS: The material is adopted easily in the operating, autogenous umbilical cord is elastic tissue and no toxicity could relax the abdominal press effectively after the operation, the survival rate is high.

[Experimental study on a novel esophageal prosthesis made of composite biomaterials].

OBJECTIVE: To design and develop a novel esophageal prosthesis by selecting appropriate biomaterials, developing special manufacturing techniques, and investigating the feasibility of replacement of cervical esophagus in mongrel dogs. METHODS: In accordance with the requirements of ideal esophageal substitutes, we designed a new type of esophageal prostheses. The inner stent were made with polyurethane of medical grade, and the outer surface of the prosthesis was coated with collagen-chitosan sponge. The silicone tube was used as a control. Thirteen adult mongrel dogs that were divided into two groups were used to establish the experimental models. RESULTS: In the experimental group (n = 8), the esophageal prostheses were completely incorporated with the native esophagus and adherent to the surrounding host connective tissues. Epithelial linings of varying degrees were formed on the luminal surface, and complete epithelization was seen in 1 month postoperatively. The granulation at the sites of the anastomosis in this group was less significant than that of the control group. One dog has been surviving for 12 months up to now without any complications. In the control group (n = 5), esophageal epithelial was not observed on the luminal surface, constriction of the regenerated esophagus progressed and all the dogs died within 2 months after operation. CONCLUSION: These observations suggest that this esophageal prosthesis made of composite biomaterials has high biocompatibility and potential for long-segment esophageal reconstruction, which is promising for the clinical repair of esophageal defects.