Association between Serum Ferritin Levels and Metabolic-associated Fatty Liver Disease in Adults: a Cross-sectional Study Based on the NHANES.

OBJECTIVE: Ferritin, initially acting as an iron-storage protein, was found to be associated with metabolic diseases. Our study was designed to investigate the association between serum ferritin and metabolic-associated fatty liver disease (MAFLD) using data from the National Health and Nutrition Examination Survey (NHANES) of the United State of America. METHODS: A cross-sectional study was conducted, enrolling a total of 2145 participants from the NHANES in the 2017-2018 cycles. Hepatic steatosis and liver fibrosis were assessed by ultrasound images and several non-invasive indexes. Multiple regression analysis was conducted to determine the associations between serum ferritin concentration and MAFLD and liver fibrosis. RESULTS: The analysis revealed that participants with higher serum ferritin levels (Q3 and Q4 groups) had a higher prevalence of MAFLD than those with the lowest serum ferritin levels [Q3 vs. Q1: OR=2.17 (1.33, 3.53), P<0.05 in fatty liver index (FLI); Q4 vs. Q1: OR=3.13 (1.91, 5.13), P<0.05 in FLI]. Additionally, participants with the highest serum ferritin levels (Q4 group) displayed a higher prevalence of liver fibrosis [Q4 vs. Q1: OR=2.59 (1.19, 5.62), P<0.05 in liver stiffness measurement; OR=5.06 (1.12, 22.94), P<0.05 in fibrosis-4 index], with significantly increased risk observed in participants with concomitant diabetes [OR=7.45 (1.55, 35.72), P=0.012]. CONCLUSION: Our study revealed that elevated serum ferritin levels are associated with a higher prevalence of MAFLD and advanced liver fibrosis in patients. Elevated serum ferritin levels combined with diabetes are important risk factors for liver fibrosis.

A simple and rapid protein purification method based on cell-surface display of SUMO-fused recombinant protein and Ulp1 protease.

The development of novel methods for highly efficient protein purification remains a research focus in the biotechnology field because conventional purification approaches, including affinity purification, gel filtration, and ion-exchange chromatography, require complex manipulation steps and are costly. Here, we describe a simple and rapid protein purification strategy in which the SUMO tag and Ulp1 protease are surface-displayed separately on Escherichia coli cells. After protein induction, the cells are harvested, resuspended in cleavage buffer, and incubated together for cleavage. In this approach, the surface-displayed Ulp1 cleaves the membrane-anchored SUMO fusion protein, resulting in the release of the target protein from the C-terminal of SUMO into the solution. The bacterial cells harboring SUMO and Ulp1 on their surfaces can be easily removed by centrifugation. To evaluate the purification method, we used red fluorescent protein (mCherry). Purified mCherry protein (7.72 ± 1.05 mg from 1 L of bacterial culture) was obtained after only 30 min of incubation. The protein purity was higher than 80%, and could be further improved (> 90%) by simple ultrafiltration. This study offers a promising and simple strategy for the purification of recombinant protein in its native form that requires only cleavage and centrifugation steps.

In vitro study of the effects of Angiostrongylus cantonensis larvae extracts on apoptosis and dysfunction in the blood-brain barrier (BBB).

It has been hypothesized that blood-brain barrier (BBB) dysfunction in Angiostrongylus cantonensis infection might be due to the apoptosis of the hosts' BBB cells. Here, we evaluated this hypothesis through several methods, all based on an in vitro mouse BBB model consisting of primary culture brain microvascular endothelial cells (BMECs) and brain astrocytic cells (BACs). In the present study, a four-hour percolation and HRP permeability experiment showed that A. cantonensis larvae extracts can increase the permeability of the BBB. Apoptosis among BMECs and BACs after exposure to larvae extracts was monitored by TUNEL and annexin-V-FITC/PI double staining. A. cantonensis larvae extracts were found to induce apoptosis in both BMECs and BACs. For this reason, we concluded that the induction of apoptosis might participate in the BBB dysfunction observed during angiostrongyliasis. Improved fundamental understanding of how A. cantonensis induces apoptosis may lead to new approaches to the treatment or prevention of this parasitic disease.

[Study on the virus-free technology of Pinellia temata].

OBJECTIVE: To solve the degradation of production and quality of Pinellia caused by the virus accumulation, rapid propagation technical of virus-free Pinellia was researched. METHODS: Pinellia leaves,petioles as explants, technology of using high temperature (38 degrees C, 40d) and shoot tip culture producing virus-free Pinellia was explored. RESULTS: The results showed that leaves without virus spots was about 88.9% when explants were culture for 40d at high temperature (38 degrees C). 1.0 mg/L 6-BA and 0.1 mg/L NAA could induce seedling from shoot tip,seedling rate is up to 91.4%; MS added 0.5 mg/L 6-BA and 0.1 mg/L NAA was conducive to growth of the plantlets; added 0.5 mg/L KT and 0.5 mg/L NAA was in favor of inducing root and promoting root growth, the survival rate of the transplanting seedling could reach 89.5%. CONCLUSION: A reliable system of virus-free Pinellia propagation is established.

[Analysis and strategy report on overseas large-scale systematic evaluation on clinical effectiveness of acupuncture].

In recent years, studies of large-scale systematic evaluation on clinical effectiveness of acupuncture were carried out in overseas. The literatures were conducted in Cochrane Library and overseas journals about systematic review of clinical effectiveness of acupuncture. The Cochrane Library contained a series of systematic reviews for the treatment of 67 kinds of diseases by acupuncture in 2009. Preliminary evaluations of clinical effectiveness of acupuncture on 37 kinds of disease were conducted. The results indicated that acupuncture therapy was effective for 7 kinds of disease, such as idiopathic headache, neck disorders, glaucoma, rheumatoid arthritis, chemotherapy-induced nausea or vomiting, primary dysmenorrhoea with TENS and knee osteoarthritis with TENS. However, these studies still need improved research designs and sufficient research evidence. The results also indicated that acupuncture was indecisive for the other 30 kinds of disease because of insufficient evidence. Through analysis, results of most systematic reviews indicated that there were no significant difference between therapeutic effects of acupuncture treatment and pseudo-acupuncture treatment. Effect of acupuncture treatment was equivalent to therapeutic effect of placebo. The likely reasons may be that some important clinical factors are disregarded in these researches, such as selection of acupoints, treatment with syndrome differentiation, the angel and depth of needle insertion, the proper time for treatment and so on. Therefore, the large-scale systematic evaluation on clinical effectiveness of acupuncture was criticized by acupuncturists. Thus, the pressing problem is to establish a rational evaluation system of clinical acupuncture. The suggestions are strengthening the research on diagnosis and treatment standard, strengthening the quality control of clinical acupuncture and establishing sound acupuncture control group and placebo acupuncture group. The basic researches on the relationship between diseases and acupoints need to be strengthened in order to explore the mechanism of acupoints reaction on diseases.

[Distribution of the activated acupoints after acute gastric mucosal injury in the rat].

OBJECTIVE: To observe the dynamic distribution of the extravasated Evans Blue (EB) dye points (neurogenic inflammatory response) at the skin after acute gastric mucosal injury (AGMI) and its relation to the related regular acupoints in the locations in rats. METHODS: A total of 70 Wistar rats were randomized into normal control (n=10), normal saline (n=10), and AGMI (n=50) groups. The AGMI group was further divided into 5 h, 2 d, 3 d, 4 d and 5 d subgroups with 10 rats in each. AGMI model was duplicated by intragastric perfusion of diluted hydrochloric acid (HCl, 0.5 mol/L). Evans Blue Dye (50 mg/kg, 50 mg/mL in 0.9% saline) was given to the rats before AGMI modeling. The plasma extravasated EB points at the skin of the whole body were observed after removal of the hair. RESULTS: The extravasated EB points presented a nerve-segmental distribution, with the proportion of the points in the location being 47.5% for "Geshu" (BL 17),58. 82% for "Jizhong" (GV 6), 88.23% for "Pishu" (BL 20), 82.35% for "Weishu" (BL21), 17.64% for "Zhongwan" (CV 12), and 5.88% for "Shangwan" (CV 13), respectively. The plasma extravasation of EB seldom appeared in normal rats and only fewer points were found in rats accepted administration of 0.9% saline. Significant differences were found between model and normal control groups, and between model and normal saline groups in the numbers of the extravasated EB points (P < 0.01, P < 0.05). The number of the extravasated EB points was related to the phase of gastric mucosa injury, being most on the 2nd and 3rd day after modeling and disappearing gradually along with the natural repair of AGMI. CONCLUSION: AGMI promotes the plasma extravasation of EB and the extravasated EB points present a nerve-segmental distribution and have a higher corresponding rate with some acupoints including "Pishu" (BL 20), "Weishu" (BL 21), etc., suggesting an activation of the normally silent acupoints under diseased conditions.

[Effect of electroacupuncture on cortical spreading depression and plasma CGRP and substance P contents in migraine rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on cortical spreading depression (CSD) and contents of plasma calcitonin gene-related peptide (CGRP) and substance P (SP) in migraine rats. METHODS: Thirty male SD rats were equally randomized into control, model and EA groups. Migraine model was established by topical application of KCI (3 mol/L) immersed in a piece of filter paper to the cerebral cortex (parietal lobe, 6 mm posterior to the Bregma and 5 mm to the sagital fissure) after exposure of the skull (in reference to Michael' method). KCI stimulation evoked CSD potentials (3 mm rostral to the Bregma, and 2 mm to the sagital fissure) were recorded by using a glass microelectrode. For rats of control group, filter paper containing 0.9% NaCl was applied to the same parietal cortex area. EA (1 mA, 2 Hz/100 Hz) was applied to bilateral "Yanglingquan" (GB 34) and "Taichong" (LR 3) for 30 min. The contents of plasma CGRP and SP were assayed by radioimmunoassay. RESULTS: CSD was induced 3-5 min after application of KCI to the parietal lobe. The average amplitude of model group was (-25.13 +/- 1.23) mV, and that of EA group was (-19.19 +/- 1.53) mV, displaying a significant reduction of CSD amplitude after EA (P < 0.01). Comparison among 3 groups showed that both plasma CGRP and SP contents in model group were significantly higher than those of control group (P < 0.001, P < 0.01), while compared with model group, plasma CGRP and SP levels in EA group decreased considerably (P < 0.05, P < 0.001), suggesting an inhibitory effect of EA on pain-producing substance. CONCLUSION: EA of GB 34 and LR 3 can effectively suppress KCI provoked cortical spreading depression and plasma CGRP and SP levels in the rat, which may contribute to its effect in relieving migraine in clinic.

[Mast cell and substance P are involved in the process of acupoint sensitization induced by acute gastric mucosal injury].

OBJECTIVE: To observe changes of mast cells (MCs) number and morphology, and substance P (SP) expression in Evans blue (EB) extravasated region around acupoint "Pishu" (BL 20) and "Weishu" (BL 21) after acute gastric mucosal injury (AGMI) so as to investigate the mechanism underlying visceral problems-induced acupoint activation. METHODS: Thirty adult Wistar rats were randomly divided into normal control (n = 15) and AGMI groups (n = 15). AGMI model was duplicated by perfusing the rats with 0.5 mol/L HCl (1 mL/100 g) after fasting for 20 h. Five hours after AGMI, the rats were treated by tail-intravenous injection of EB dye (5 mg/100 g, 50 mg/mL in normal saline) for inducing dye-plasma extravasation in the skin around BL 20, BL 21 regions, etc. at the back. The rats of the normal control group were treated with tail-intravenous injection of 0.9% NaCl. The skin and subcutaneous tissues (2 mmx 2 mm) of extravasated EB dye points (BL 20 or BL 21 region) and those 2 mm lateral to the extravasated EB dye points in the model group and the corresponding points in the normal control group were sampled (followed by fixing them in 4% paraformaldehyde), sectioned and stained by toluidine blue (for labeling MCs). The expression of SP in the extravasated EB dye skin and subcutaneous tissues was detected by immunohistochemistry (n = 5) and western blot (n = 5) respectively. The number of MCs in these samples was counted and the degranulation rate of MCs calculated. RESULTS: The total number of MCs and the number of degranulated MCs were significantly more in the EB extravasation points (corresponding to BL 20/BL 21 area) of AGMI group than those in the control spots of AGMI group and than those in the normal control group (P < 0.05, P < 0.001). The degranulation rate of MCs was significantly higher in the EB extravasation points of AGMI group than those in the control spots of AGMI group and in the normal control group (P < 0.01). In comparison with normal control group, the SP expression level was increased consideraly in the control spots of AGMI group and AGMI group (P < 0.01). CONCLUSION: After AGMI, the numbers of MCs and the degranulated MCs, and the SP expression level in BL 20/BL 21 area were increased significantly, suggesting an involvement of MCs and SP in the process of AGMI-induced activation of acupoints.