[The research of liver failure in Banna miniature pigs caused by amanita exitialis].

Objective: To explore the characteristics of Banna miniature pig liver failure induced by amanita exitialis. Methods: From September to October 2020, a reverse high performance liquid chromatography (RP-HPLC) method was used to determine the toxin content of amanita exitialis solution, and 2.0 mg/kg amanita exitialis solution (α-amanitins+ß-amanitins) was administered orally to Banna miniature pigs. Toxic symptoms, blood biochemical indexes and histopathological changes of liver, heart and kidney were observed at each time point. Results: All Banna miniature pigs died within 76 h of exposure, and different degrees of digestive tract symptoms such as nausea, vomiting and diarrhea appeared between 6 and 36 h. The biochemical indexes of alanine aminotransferase, aspartate aminotransferase, total bilirubin, lactate dehydrogenase, myoglobin, creatine kinase isoenzyme, blood urea nitrogen and creatinine increased significantly at 52 h after exposure, and the differences were statistically significant compared with 0 h (P<0.05). The bleeding of liver and heart was obvious under macroscopic and microscopic observation, hepatocyte necrosis, renal tubule epithelial cell swelling. Conclusion: Large dose of amanita exitialis can cause acute liver failure of Banna miniature pigs, which is in line with the pathophysiological characteristics of acute liver failure, and lays a foundation for further research on the toxic mechanism and detoxification drugs of amanita exitialis induced liver failure.

[Molecular epidemiology and clinical characteristics of six cases of CA-MRSA pneumonia after influenza].

Objective: To summarize and analyze the strains' molecular epidemiology and clinical characteristics of 6 strains of post-influenza community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) pneumonia. Methods: Six cases of CA-MRSA pneumonia after influenza from 2014 to 2022 were retrospectively collected and CA-MRSA strains from each patient were cultured. Then, SCCmec typing, MLST typing, and spa typing were performed on the samples, which also included the procedures for the detection of virulence factors. Antibiotic susceptibility test was then performed on all 6 strains. Results: ST59-t437-Ⅳ was the predominant type in all the strains of CA-MRSA(2/6). Leukocidin (PVL) was detected in 5 cases, and hemolysin α (HLAα) and phenol soluble regulatory protein α (PSMα) were detected in 6 cases. Five of the cases included in this study were diagnosed with severe pneumonia. In terms of treatment, 4 cases received antiviral therapy, and 5 patients with severe pneumonia received anti-infection treatment with vancomycin as the first choice and were discharged after improvement of their condition. Conclusions: The molecular types and virulence factors of CA-MRSA after influenza infection could vary considerably. Our experiments also showed that secondary CA-MRSA infection after influenza was more common in young people with no underlying diseases and could cause severe pneumonia. Vancomycin and linezolid were the first-line drugs for treating CA-MRSA infection and were highly effective in improving the condition of diagnosed patients. We highlighted the importance of referring patients with severe pneumonia after influenza for etiological tests to determine whether they had CA-MRSA infection, so that they could be properly treated with anti-influenza agents and receive appropriate anti-CA-MRSA infection treatment.

[Investigation on a case of Amanita neoovoidea poisoning].

This paper reported a case of poisoning caused by ingestion of Amanita neoovoidea. The patient experienced nausea, vomiting, oliguria, acute renal function injury, and was discharged after symptomatic support treatment and blood purification treatment. Given the different toxicity of different mushrooms, species identification of poisonous mushrooms can help clinicians in diagnosis and treatment.

[Three cases of mushroom poisoning caused by lethal Amanita species].

Mushroom poisoning with amatoxins can cause liver dysfunction in patients, and death in severe cases. The amatoxins detection by enzyme-linked immunosorbent assay (ELISA) can help early clinical diagnosis. Three patients were identified as α-amatoxin containing mushroom poisoning by ELISA. The first symptoms of patients was gastrointestinal symptoms, and liver function damage occured later. One patient gave up treatment and died. After received supportive treatments such as adsorption of toxins, catharsis, fluid supplementation to promote toxin metabolism and liver protection, 2 patients were recovered and discharged.

Prognostic potentials of miRNA-19a-3p and PDCD5 in nasopharynx carcinoma.

OBJECTIVE: To determine expressions of MicroRNA-19a-3p (miRNA-19a-3p) and PDCD5 in nasopharyngeal carcinoma (NPC) tissues, and their prognostic potentials in NPC. PATIENTS AND METHODS: Expressions of miRNA-19a-3p and PDCD5 in NPC tissues and controls were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between expressions of miRNA-19a-3p and PDCD5 in NPC was evaluated by Pearson correlation test. Furthermore, potential influences of miRNA-19a-3p and PDCD5 on clinical features of NPC patients were assessed. Through 5-year follow-up, survival analysis in NPC patients was conducted by Kaplan-Meier method. Finally, factors influencing prognosis of NPC were determined using the Cox regression model. RESULTS: MiRNA-19a-3p was upregulated and PDCD5 was downregulated in NPC tissues. Pearson correlation test uncovered a negative correlation between expression levels of miRNA-19a-3p and PDCD5 in NPC tissues. MiRNA-19a-3p level was correlated with N classification and clinical stage in NPC patients, while PDCD5 level was correlated with T classification, pathological grade and clinical stage. Survival analysis showed poor prognosis in NPC patients expressing high level of miRNA-19a-3p or low level of PDCD5. Cox regression analysis illustrated that N2+3 classification, clinical stage III+IV, high level of miRNA-19a-3p and low level of PDCD5 were independent risk factors for the prognosis of NPC. CONCLUSIONS: MiRNA-19a-3p is upregulated and PDCD5 is downregulated in NPC tissues. High level of miRNA-19a-3p and low level of PDCD5 are unfavorable for the prognosis of NPC.

Methylation-Based Age Estimation Model Construction and Its Effectiveness Evaluation.

OBJECTIVES: To screen the DNA methylation loci associated with the age of Han males in northern China and to construct an age estimation model. METHODS: Twenty-one candidate methylation loci were screened. The DNA methylation levels of 476 blood samples from Chinese Han males were detected for 21 amplicons using EpiTYPER technology platform, and data on 153 DNA methylation loci were obtained. RESULTS: After correlation analysis, 8 age-related DNA methylation loci were finally screened. CpG1, CpG2, CpG4, CpG7, CpG8 were located on TRIM59, RASSF5, Clorf132, CSNK1D, ELOVL2,CpG5, CpG6 on PDE4C, and CpG3 on chr17:21452808. Based on the 8 loci, 352 samples were used for model construction. A multivariate linear regression age estimation model was constructed （R2=0.93）, with mean absolute deviation （MAD） of 2.69 years old. When 109 samples were used for model validation, the MAD was 3.80 years old. The test was repeated 3 times in 15 new samples, with MADs of 4.08, 4.68 and 3.93 years old, respectively. CONCLUSIONS: The age estimation model on Han males in northern China constructed in this study can be used to estimate the age of victims and suspects and to narrow the scope of investigation, and therefore has practical application value.

Risk factors for imipenem-resistant Pseudomonas aeruginosa in neonatal intensive care units in south China.

This multi-centre retrospective study was designed to investigate the risk factors for infection with imipenem-resistant Pseudomonas aeruginosa in neonatal intensive care units (NICUs) in south China. All patients with confirmed P. aeruginosa infection from eight NICUs in south China were divided into two groups: imipenem-susceptible P. aeruginosa and imipenem-resistant P. aeruginosa. Data were analysed using Chi-squared test and logistic regression. In total, 188 medical records were reviewed. On multi-variate logistic analysis, the only independent risk factor was imipenem treatment within two weeks of isolation of P. aeruginosa (odds ratio 6.409, 95% confidence interval 1.926-21.333).

Response to transarterial chemoembolization as a selection criterion for resection of hepatocellular carcinomas.

BACKGROUND: Liver resection for intermediate (Barcelona Clinic Liver Cancer (BCLC) stage B) hepatocellular carcinoma (HCC) remains controversial. This study attempted to demonstrate the effectiveness of preresection transarterial chemoembolization (TACE) as a selection criterion for BCLC-B HCC. METHODS: The study included patients with BCLC-B HCC who underwent liver resection after TACE. The tumour response to TACE was evaluated according to the modified Response Evaluation Criteria in Solid Tumours (mRECIST). Patients with a complete or partial response comprised the responder group, whereas those with stable or progressive disease were classified as non-responders. RESULTS: A total of 242 patients were included. After between one and eight sessions of TACE, 141 patients were included in the responder group: 37 patients (15·3 per cent) who achieved a complete response and 104 who had a partial response. The cumulative 1-, 3- and 5-year overall survival rates were 97·2, 88·7 and 75·2 per cent respectively in the responder group, compared with 90·1, 67·3 and 53·5 per cent among 101 non-responders (P < 0·001). Tumour-free survival rates were also better among responders than non-responders (P < 0·001). In multivariable analysis, independent predictors of overall and tumour-free survival were response to TACE and microvascular invasion (all P < 0·001). CONCLUSION: mRECIST may represent selection criterion for intermediate HCC for surgical treatment.

Tauroursodeoxycholate, a chemical chaperone, prevents palmitate-induced apoptosis in pancreatic ß-cells by reducing ER stress.

BACKGROUND: Free fatty acids (FFA) can have deleterious effects on ß-cells and promote type 2 diabetes, a process known as lipotoxicity. Recently, the induction of endoplasmic reticulum (ER) stress is one mechanism proposed to contribute to the detrimental effects of FFA on ß-cells. Tauroursodeoxycholic acid (TUDCA) has been reported to show cytoprotective effects by alleviating ER stress induced by some cytotoxic stimuli. The aim of this study was to investigate the effects of TUDCA on FFA (palmitate)-induced apoptosis and ER stress in rat islet ß-cells. METHODS: The rat pancreatic ß-cell line INS-1 was cultured with palmitate (0.5 mM), or cultured togther with TUDCA (100 µM), Annexin V-fluorescein-isothiocyanate/propidium iodide flow cytometry was used to assess apoptosis in INS-1 cells. Cell viability was evaluated with MTT reduction conversion assay. The expressions of ER stress marker GRP78, ER stress-associated pro-apoptotic effectors CHOP and ATF4 were detected by Western blotting. RESULTS: TUDCA significantly reduced palmitate-induced cell apoptosis and growth inhibition in INS-1 cells. TUDCA also attenuated palmitate-induced expressions of GRP78, CHOP and ATF4 in INS-1 cells. CONCLUSIONS: Our results thus suggested that TUDCA could protect INS-1 cells from palmitate-induced injury, which might be due to the amelioration of ER stress and blocking the ATF4/CHOP signaling pathway.

Inhibition of tumor cell proliferation and induction of apoptosis in human lung carcinoma 95-D cells by a new sesquiterpene from hairy root cultures of Artemisia annua.

Artemisia annua is a rich source of many bioactive substances, and in our recent work, a new sesquiterpene, (Z)-7-acetoxy-methyl-11-methyl-3-methylene-dodeca-1,6,10-triene (AMDT), was isolated and identified from hairy roots culture of A. annua, and its bioactivity was characterized in this work. AMDT showed moderate cytotoxic activities against the human tumor cell lines of HO8910 (ovary), 95-D (lung), QGY (liver) and HeLa (cervix) by MTT assay, whose IC(50) values were ranged within 52.44-73.3 microM. As lung cancer is the No. 1 killer of global cancer patients, our interest is to investigate the ability of AMDT in inducing apoptosis of 95-D tumor cells. The 95-D cell growth was inhibited by AMDT, and the flow cytometry analysis showed its cell cycle was arrested in the G1 phase. The apoptotic rate of the cells increased in a dose-dependent manner. AMDT lowered the mitochondrial membrane potential and increased the expression of caspase-9 and -3. These results revealed that AMDT could efficiently induce 95-D cell apoptosis through mitochondrial dependent pathway, and it may be a potential chemotherapeutic agent.

Damage of Escherichia coli membrane by bactericidal agent polyhexamethylene guanidine hydrochloride: micrographic evidences.

AIMS: The purpose of this study was to provide micrographic evidences for the damaged membrane structure and intracellular structure change of Escherichia coli strain 8099, induced by polyhexamethylene guanidine hydrochloride (PHMG). METHODS AND RESULTS: The bactericidal effect of PHMG on E. coli was investigated based on beta-galactosidase activity assay, fluorescein-5-isothiocyanate confocal laser scanning microscopy, field emission scanning electron microscopy and transmission electron microscopy. The results revealed that a low dose (13 microg ml(-1)) of PHMG slightly damaged the outer membrane structure of the treated bacteria and increased the permeability of the cytoplasmic membrane, while no significant damage was observed to the morphological structure of the cells. A high dose (23 microg ml(-1)) of PHMG collapsed the outer membrane structure, led to the formation of a local membrane pore across the membrane and badly damaged the internal structure of the cells. Subsequently, intracellular components were leaked followed by cell inactivation. CONCLUSIONS: Dose-dependent membrane disruption was the main bactericidal mechanism of PHMG. The formation of the local membrane pores was probable after exposure to a high dose (23 microg ml(-1)) of PHMG. Micrographic evidences were provided about the damaged membrane structure and intracellular structure change of E. coli. SIGNIFICANCE AND IMPACT OF THE STUDY: The presented information helps understand the bactericidal mechanism of PHMG by membrane damage.

Every-other-day interferon beta-1b versus once-weekly interferon beta-1a for multiple sclerosis (INCOMIN Trial) II: analysis of MRI responses to treatment and correlation with Nab.

BACKGROUND: In RRMS, clinical exacerbations are usually associated with different types of active lesions at MRI, including: hyperintense lesions on T1-weighted post-gadolinium sequences; new hyperintense lesions or enlarging old lesions on PD/T2-weighted scans; or new hypointense lesions on T1-weighted pre-Gd sequences. OBJECTIVE/METHODS: Primary outcome was the occurrence of patients with at least one active MRI lesion of the different types indicated above during treatment with 250 microg every other day (EOD) interferon beta (IFNbeta)-1b or 30 microg once weekly (OW) IFNbeta-1a in outpatients with RRMS (INCOMIN Trial). RESULTS: The number of patients with at least one 'active' lesion, evaluated over the two-year follow-up, was significantly (P = 0.014) lower in the EOD IFNbeta-1 b arm (1 3/76, 17%) then in the OW IFNbeta-1a arm (25/73, 34%). NAb frequency over two-year follow-up was 22/65 (33.8%) in the EOD IFNbeta-1b arm and 4/62 (6.5%) in the OW IFNbeta-1a arm, significantly greater in the EOD IFNbeta-1b arm. CONCLUSIONS: The development of MRI active lesions is strongly reduced by EOD-IFNbeta-1b compared with OW-IFNbeta-1a, indicating that EOD-IFNbeta-1b is more effective than OW-IFNbeta-1a in reducing ongoing inflammation and demyelination in MS. Logistic regression showed that NAb status did not affect the risk of MRI activity.

Biochemical engineering of the production of plant-specific secondary metabolites by cell suspension cultures.

Plant cell culture has recently received much attention as a useful technology for the production of valuable plant-derived secondary metabolites such as paclitaxel and ginseng saponin. The numerous problems that yet bewilder the optimization and scale-up of this process have not been over emphasized. In spite of the great progress recorded in recent years towards the selection, design and optimization of bioreactor hardware, manipulation of environmental factors such as medium components, light irradiation, shear stress and O2 supply needs detailed investigations for each case. Recent advances in plant cell processes, including high-density suspension cultivation, continuous culture, process monitoring, modeling and control and scale-up, are also reviewed in this chapter. Further developments in bioreactor cultivation processes and in metabolic engineering of plant cells for metabolite production are expected in the near future.

Improvement of Panax notoginseng cell culture for production of ginseng saponin and polysaccharide by high density cultivation in pneumatically agitated bioreactors.

A Panax notoginseng cell culture was successfully scaled up from shake flask to 1.0-L bubble column reactor and concentric-tube airlift reactor. High-density bioreactor batch cultivation was carried out using a modified MS medium. The maximum cell density in batch cultures reached 20.1, 21.0 and 24.1 g/L in the shake flask, bubble column and airlift reactors, respectively, and their corresponding biomass productivity was 950, 1140 and 1350 mg/(L x d) for each. The productivity of ginseng saponin was 70, 96 and 99 mg/(L x d) in the flask, bubble column and airlift reactors, respectively; and the polysaccharide productivity reached 104, 119 and 151 mg/(L x d) for each. Furthermore, a fed-batch cultivation strategy was developed on the basis of specific oxygen uptake rate (SOUR), i.e., sucrose feeding before a sharp decrease of SOUR, and the highest cell density of 29.7 g/L was successfully achieved in the airlift bioreactor on day 17 with a very high biomass productivity of 1520 mg/(L x d). The concentrations of ginseng saponin and polysaccharide reached about 2.1 and 3.0 g/L, respectively, and their productivity was 106 (saponin) and 158 mg/(L x d) (polysaccharide). This work successfully demonstrated the high-density bioreactor cultivation of P. notoginseng cells in pneumatically agitated bioreactors and the reproduction of the shake flask culture results in bioreactors. The cell density, biomass productivity, production titer and productivity of both ginseng saponin and polysaccharide obtained here were the highest that have been reported on a reactor scale for all the ginseng species.

Diffusion-weighted MR in reversible wernicke encephalopathy.

Diffusion-weighted images (DWI) of a patient with Wernicke encephalopathy were obtained during routine MR examination. Mammillary bodies were hyperintense on T2-weighted and enhanced on T1-weighted images; on DWI, a mild hyperintensity was noticed. Calculation of the apparent diffusion coefficient (ADC) demonstrated an increased diffusion on the affected regions; the hyperintensity on DWI was probably due to a "T2-shine-through" effect. These findings are consistent with the presence of extracellular oedema, without significant neuronal damage. The patient recovered promptly after thiamine administration, and MR alterations disappeared. The favourable evolution indicates that no relevant neuronal death occurred. This is consistent with DWI findings. DWI are more sensitive than ordinary T1- and T2-weighted images to neuronal irreversible damage, and may differentiate between neuronal necrosis and extracellular oedema in various brain pathologies. The demonstration of a limited neuronal damage may represent a favourable prognostic factor in patients with WE.

Improvement of the performance of H2O2 oxidation at low working potential by incorporating TTF-TCNQ into a platinum wire electrode for glucose determination.

A micro-biosensor was constructed by incorporating the organic conducting salt tetrathiafulvalene-tetracyanoquinodimethane (TTF-TCNQ) into a platinized platinum (Pt) wire and further covering with the electrochemical polymerical heteropolypyrrole film, in which glucose oxidase (GOx) was entrapped. The enzyme electrode can sensitively determine glucose at a low working potential, mainly based on the oxidation of H2O2. The incorporated TTF-TCNQ can significantly improve the oxidation of H2O2 on the electrode, although a part of the TTF-TCNQ functions as a mediator. Compared with the same electrode prepared without TTF-TCNQ incorporated, the TTF-TCNQ modified electrode had better performance characteristics at a working potential of 200 mV (versus SCE). The response time to 90% of the steady value was shortened from about 40 s to less than 10 s, the lower limit of the linear response was greatly extended from about 1.6 mM to 10 microM, the linear range was shifted from 1.6-10.0 to 0.01-5 mM and the sensitivity was increased from about 1 to 1.5 microA/mM. The electrode was quite stable. For continuous operation, the electrode could work for about 5 weeks and only lost 60% of its original sensitivity. Stored at 4 degrees C for intermittent determinations, the electrode kept 80% sensitivity for over 6 months. Due to covering the electrode with a non-conductive heteropolypyrrole film, ascorbate, urate and 4-acetamidophenol caused only negligible current response at an applied potential of 200 mV.

Production of ginseng and its bioactive components in plant cell culture: current technological and applied aspects.

Ginseng (the root of Panax ginseng CA Mayer) is a valuable oriental herb, which has been used in traditional Chinese medicine for thousands of years, both as a disease-healing drug and a general tonic. The medicinal value of ginseng is now also widely recognized in the west and the world ginseng market is expanding. The current supply of ginseng depends mainly on field cultivation, which is a slow and laborious process. Plant cell and tissue culture methods have been explored as potentially more efficient alternatives for the mass production of ginseng and its bioactive components. Research into ginseng cell and tissue cultures started in the early 1960s and commercial applications have been underway since the late 1980s. The ginseng cell culture has continued to attract considerable research and development effort in recent years as scientists seek to understand and optimize the culture conditions. In this paper, we review recent studies on ginseng cell culture processes, focusing on the physiological and bioengineering factors affecting the productivity of ginseng biomass and useful metabolites (e.g. ginseng saponin and polysaccharide) and the progress and concerns in large-scale applications.