Investigation and molecular identification of Anisakis infection in marine fish sold in Fuxin, Liaoning Province / 中国热带医学

@#Abstract: Objective　To investigate the infection of Anisakis in marine fish sold in Fuxin, and conduct molecular identification and evolutionary tracing of third-stage larvae to determine Anisakis species. Methods　From 2018 to 2021, marine fish sold in the market were collected randomly, and the third stage larvae of Anisakis were detected in marine fish sold in the market by direct dissection, and the morphological characteristics were used to preliminarily identify species by microscopy; the total DNA was extracted, the internal transcribed spacer sequence of the ribosomal DNA of Anisakis was amplified, and the sequence alignment and evolution analysis were carried out. Results　A total of 289 market-sold sea fish samples of marine fish sold in the market were dissected and 84 samples of Anisakis were detected with a detection rate of 29.1%, of which the infection rates of hairtail and small yellow croaker were higher, at 41.4% and 41.2%, respectively. BLAST comparison of 28 sequences revealed eight species of anisakids, including Anisakis pegreffii, Anisakis simplex, Anisakis typical, Raphidascaris trichiurid, Contracaecum muraenesoxi, Hysterothylcaium zhoushanensis, Hysterothylacium amoyense and Hysterothylcaium fabri,belonging to the genera Anisakis and Hysterothylacium. The phylogenetic tree constructed from 28 sequences generally formed two topological branches, with Anisakis pegreffii, Anisakis simplex, and Anisakis typical forming three separate clusters as the topology branch of Anisakis genus. However, meanwhile, Hysterothylacium, Contracaecum, and Raphidascaris formed a separate topological branch. Conclusions　The marine fish sold in Fuxin City have severe anisakid infection, with a wide variety of anisakid species, the dominant species being Anisakis pegreffii.

Mediating effect analysis of visceral adiposity index on free triiodothyronine to free thyroxine ratio and non-alcoholic fatty liver disease in euthyroid population.

Background: The association between free triiodothyronine/free thyroxine (FT3/FT4) and non-alcoholic fatty liver disease (NAFLD) in euthyroid subjects is unclear. In addition, few studies have explored whether VAI mediates the association between FT3/FT4 ratio and NAFLD in the euthyroid population. We aimed to analyze the mediating effect of VAI on the FT3/FT4 ratio and NAFLD risk in the euthyroid population. Methods: This cross-sectional study included 7 946 annual health examinees from the Health Examination Center, Hebei General Hospital, from January to December 2020. The basic information and biochemical parameters, as well as calculated FT3/FT4 ratio and VAI were collected. NAFLD was diagnosed according to abdominal ultrasonography. The fibrosis score for NAFLD positive subjects (NFS) was calculated to reflect the extent of liver fibrosis. The risk of NAFLD was analyzed by quartiles of FT3/FT4 ratio (Q1-Q4 quartiles) and VAI (V1-V4 quartiles), respectively. Pearson correlation analysis was performed to investigate the correlation between FT3/FT4 ratio and VAI. Multivariate logistic regression analysis was applied to analyze the effect of FT3/FT4 ratio and VAI on NAFLD and NFS status. Bootstrap was conducted to explore whether VAI mediated the association between FT3/FT4 ratio and NAFLD. Results: Of the 7 946 participants, 2 810 (35.36%) had NAFLD and 5 136 (64.64%) did not. Pearson correlation analysis indicated that FT3/FT4 ratio was positively associated with VAI (P<0.05). Multivariate logistic regression analysis indicated that compared to the Q1 group, the risk of NAFLD significantly increased in Q3 group [OR=1.255, 95%CI (1.011, 1.559)] and Q4 group [OR=1.553, 95%CI (1.252, 1.926)](P<0.05). Compared to the V1 group, the risk of NAFLD notably increased in V2 group [OR=1.584, 95%CI (1.205, 2.083)], V3 group [OR=2.386, 95%CI (1.778, 3.202)] and V4 group [OR=4.104, 95%CI (2.835, 5.939)] (P<0.01). There was no relevance between FT3/FT4 ratio, VAI and NFS status. Mediating effect analysis showed that FT3/FT4 ratio significantly directly influenced NAFLD prevalence [ß=3.7029, 95%CI (2.9583, 4.4474)], and VAI partly mediated the indirect effect of the FT3/FT4 ratio on NAFLD prevalence [ß=2.7649, 95%CI (2.2347, 3.3466)], and the mediating effect accounted for 42.75% of the total effects. Conclusion: Both FT3/FT4 ratio and VAI were predictors of NAFLD, and VAI partly mediated the indirect effect of the FT3/FT4 ratio on NAFLD prevalence in the euthyroid population.

A platform for blue-luminescent carbon-centered radicals.

Organic radicals, which have unique doublet spin-configuration, provide an alternative method to overcome the efficiency limitation of organic light-emitting diodes (OLEDs) based on conventional fluorescent organic molecules. Further, they have made great breakthroughs in deep-red and near-infrared OLEDs. However, it is difficult to extend their fluorescence into a short-wavelength region because of the natural narrow bandgap of the organic radicals. Herein, we significantly expand the scope of luminescent radicals by showing a new platform of carbon-centered radicals derived from N-heterocyclic carbenes that produce blue to green emissions (444-529 nm). Time-dependent density functional theory calculations and experimental investigations disclose that the fluorescence originates from the high-energy excited states to the ground state, demonstrating an anti-Kasha behavior. The present work provides an efficient and modular approach toward a library of carbon-centered radicals that feature anti-Kasha's rule emission, rendering them as potential new emitters in the short-wavelength region.

Treatment of gastric carcinoma with lymphoid stroma by immunotherapy: A case report.

BACKGROUND: Gastric cancer with lymphoid stroma (GCLS) is a rare type of gastric cancer characterized by abundant lymphocytic infiltration of the stroma. It is an Epstein-Barr virus-associated gastric cancer with a better prognosis than typical gastric cancer but with similar symptoms. GCLS diagnosis is based on pathological, histological and immunohistochemical examination and there are no standardized guidelines for treatment. CASE SUMMARY: This case report describes a 72-year-old man with a 6-mo history of abdominal pain. Endoscopy revealed ulcerative lesions in the stomach and gastric cancer was suspected. A preoperative endoscopic biopsy indicated undifferentiated carcinoma and postoperative pathological, histological and immunohistochemical analyses of the resected specimen confirmed a final diagnosis of GCLS. CONCLUSION: The patient showed high programmed cell death-ligand 1 expression and recovered well after immunotherapy.

Predictable maternal separation confers adult stress resilience via the medial prefrontal cortex oxytocin signaling pathway in rats.

Early-life stress is normally thought of as a major risk for psychiatric disorders, but many researchers have revealed that adversity early in life may enhance stress resilience later in life. Few studies have been performed in rodents to address the possibility that exposure to early-life stress may enhance stress resilience, and the underlying neural mechanisms are far from being understood. Here, we established a "two-hit" stress model in rats by applying two different early-life stress paradigms: predictable and unpredictable maternal separation (MS). Predictable MS during the postnatal period promotes resilience to adult restraint stress, while unpredictable MS increases stress susceptibility. We demonstrate that structural and functional impairments occur in glutamatergic synapses in pyramidal neurons of the medial prefrontal cortex (mPFC) in rats with unpredictable MS but not in rats with predictable MS. Then, we used differentially expressed gene (DEG) analysis of RNA sequencing data from the adult male PFC to identify a hub gene that is responsible for stress resilience. Oxytocin, a peptide hormone, was the highest ranked differentially expressed gene of these altered genes. Predictable MS increases the expression of oxytocin in the mPFC compared to normal raised and unpredictable MS rats. Conditional knockout of the oxytocin receptor in the mPFC was sufficient to generate excitatory synaptic dysfunction and anxiety behavior in rats with predictable MS, whereas restoration of oxytocin receptor expression in the mPFC modified excitatory synaptic function and anxiety behavior in rats subjected to unpredictable MS. These findings were further supported by the demonstration that blocking oxytocinergic projections from the paraventricular nucleus of the hypothalamus (PVN) to the mPFC was sufficient to exacerbate anxiety behavior in rats exposed to predictable MS. Our findings provide direct evidence for the notion that predictable MS promotes stress resilience, while unpredictable MS increases stress susceptibility via mPFC oxytocin signaling in rats.

Characterization of the Salmonella enterica Serotype Isangi Isolated from Patients for the First Time in China.

No studies have reported the isolation of serotype Salmonella Isangi from cases of salmonellosis in mainland China. We investigated an outbreak of foodborne disease with salmonella and collected the samples from the patients and surplus foods. Salmonella strains were isolated and the serotype was identified according to the Kauffmann-White scheme. The relatedness of the isolates was determined using pulsed-field gel electrophoresis (PFGE) and whole genome sequencing (WGS). Antimicrobial susceptibility was conducted by the broth microdilution method. There were 74 diners in the case, 33 of which got ill, with an attack rate of 44.6% (33/74). A total of 24 samples were collected from the outbreak cases, six Salmonella Isangi strains were isolated and susceptible to all tested drugs. PFGE and WGS analysis suggested that the pathogen dissemination through a single or limited vector(s), the steamed fish and mixed food (fry spicy chicken, braised pork ribs, and goose leg), may be the source of infection or be cross-contaminated. We first report the characteristics of an outbreak and molecular strain relatedness of Salmonella Isangi in mainland China.

Orexin-A-induced ERK1/2 activation reverses impaired spatial learning and memory in pentylenetetrazol-kindled rats via OX1R-mediated hippocampal neurogenesis.

Epilepsy is characterized by the occurrence of repetitive seizures and can greatly affect a patient's cognition, particularly in terms of learning and memory. Orexin-A is an excitatory neuropeptide produced by the lateral hypothalamus that has been shown to be involved in learning and memory. A reduction in the levels of orexin-A after seizures may underlie the learning and memory impairments induced by epilepsy. Thus, we used pentylenetetrazol (PTZ)-kindled rats to investigate the effects of orexin-A on learning and memory and the involvement of neurogenesis in the dentate gyrus in OX1R-mediated ERK1/2 activation. A Morris water maze test revealed reduced escape latencies, prolonged times in the target quadrant and an increased number of platform crossings in PTZ-kindled rats exposed to orexin-A. These ameliorating effects of orexin-A on spatial learning and memory were attenuated by the intracerebroventricular injection of the OX1R antagonist SB334867 or the ERK1/2 inhibitor U0126. Further studies using bromodeoxyuridine (BrdU) revealed that orexin-A increased the number of BrdU-positive cells, doublecortin (DCX)/BrdU levels and the number of NeuN/BrdU double-positive nuclei in the dentate gyrus of PTZ-kindled rats. However, these effects were inhibited by treatment with SB334867 or U0126. Taken together, these data suggest that orexin-A attenuated the impairment of spatial learning and memory in PTZ-kindled rats and that this attenuation involved neurogenesis in the dentate gyrus via OX1R-mediated ERK1/2 activation.

Risk factors associated with severe manifestations of 2009 pandemic influenza A (H1N1) infection in China: a case-control study.

BACKGROUND: No studies on the risk factors of 2009 pandemic influenza A (H1N1) in China have been reported. We aimed to investigate the risk factors for severe manifestations of 2009 pandemic H1N1 influenza in China METHODS: A case-control study with 343 severe hospitalized patients and 343 randomly selected mild controls was conducted. The diagnosis was established by assessment of clinical symptoms and confirmed by the real-time reverse-transcriptase-polymerase chain reaction assay. Severe or mild patients were classified by uniform criteria issued by the Ministry of Health in China. RESULTS: The multivariable logistic regression analysis showed that the overweight or obese subjects admitted to hospital with H1N1 influenza were more likely to experience severe manifestations. The ORs were 3.70 (95% CI: 2.04-6.72) and 35.61 (95% CI: 7.96-159.21) respectively. Subjects at age less than 5 years or older than 60 years had an increased risk of severe manifestations (OR = 21.14, 95% CI: 7.79-57.33). We also observed increased risk among subjects with longer time interval from symptom onset to hospital admission (OR = 3.26, 95% CI: 2.08-5.11) or peasants (OR = 9.79, 95% CI: 5.11-18.78). Those with chronic disorders had increased risk of severe manifestations of H1N1 influenza. CONCLUSION: We provide evidence on the risk factors associated with severe manifestations of 2009 pandemic H1N1 influenza in a study of hospitalized subjects in China.

[Effects of Chinese herbal medicine Xifeng Capsule on multidrug resistance-associated protein 1 expression in hippocampus and cortex of rats with lithium-pilocarpine-induced epilepsy].

OBJECTIVE: To study the effects of Xifeng Capsule, a compound traditional Chinese herbal medicine, combined with carbamazepine on spontaneous epileptic seizure induced by lithium and pilocarpine in rats and the expression level of multidrug resistance-associated protein 1 (MRP1). METHODS: Lithium and pilocarpine were used to induce epilepsies in rats. All epileptic rats were randomly divided into model, high-dose Xifeng Capsule, medium-dose Xifeng Capsule, low-dose Xifeng Capsule, high-dose Xifeng Capsule plus carbamazepine (CBZ) (combined high-dose group), high-dose Xifeng Capsule plus half dose of CBZ (combined low-dose group) and CBZ groups with 10 rats in each group. And another 10 normal rats served as control. After treating 28 d, immunohistochemical method was used to detect the MRP1 expression in cortex and hippocampus of the epileptic rats. RESULTS: MRP1 expression in hippocampus of the treated groups was higher than that of the normal control group, with wider range and darker positive particles, but was lower than that of the model group. In the cortical areas, the differences between the combined high-dose group or the combined low-dose group and the model group were statistically significant (P<0.05). Regardless of the hippocampus CA1, CA3, gyrus or cortical areas, the influence of high-dose Xifeng Capsule on MRP1 distribution was superior to that of low-dose Xifeng Capsule; Xifeng Capsule combined with CBZ had better effects than low-dose Xifeng Capsule, medium-dose Xifeng Capsule and CBZ used alone (P<0.05). CONCLUSION: Xifeng Capsule used alone or combined with CBZ can effectively inhibit MRP1 expression in hippocampus and cortex of epileptic rats.

[Reversal effect of gambogic acid on multidrug resistance of K562/A02 cell line].

This study was purposed to investigate the reversal effect of gambogic acid (GA) on multidrug resistance of K562/A02 cells and its mechanism. The IC(50) (half maximal inhibitory concentration) of adriamycin (ADM) was evaluated by MTT. Cell apoptosis was detected by flow cytometry. Morphological changes of K562/A02 cells were observed by fluorescent microscopy with DAPI staining. The expressions of Survivin and P-gp were determined by Western blot. The results showed that the IC(50) of ADM on K562 and K562/A02 cell proliferation were (1.42 ± 0.07) µg/ml and (28.42 ± 1.40) µg/ml respectively. GA ≤ 0.0625 µmol/L had no inhibitory effect on proliferation of K562 and K562/A02. 0.0625 µmol/L GA could enhance the sensitivity of K562/A02 cells to ADM (P < 0.05) and the reversal multiples was 1.53. The apoptotic rate was raised after treating with ADM combined with 0.0625 µmol/L GA for 48 h (P < 0.05). Morphological differences were typical and obvious between cells of control and treated groups under fluorescence microscopy using DAPI staining. After treating K562/A02 cells with ADM combined with 0.0625 µmol/L GA for 48 h, the expressions of Survivin and P-gp were down-regulated at protein levels. It is concluded that GA can enhance the sensitivity of K562/A02 cells to ADM, which may be related to increasing cell apoptosis and down-regulating expressions of Survivin and P-gp.