Phyllosticta rizhaoensis sp. nov. causing leaf blight of Ophiopogon japonicus in China.

Ophiopogon japonicus (Asparagaceae) is a perennial grass species which can be cultivated as an ornamental and medicinal plant. From April 2021 to September 2022, a serious leaf blight disease of O. japonicus was discovered in Rizhao City, Shandong Province, China. The initial disease symptoms were small yellow spots, finally developing as tip blight, often associated with many small, black, semi-immersed pycnidial conidiomata formed in lesions. To obtain isolates of the causal agent for this disease, samples were randomly collected from O. japonicus diseased leaves in Rizhao City. In total 97 Phyllosticta isolates were obtained from samples, and studied using morphological features and multi-locus phylogenetic analyses of a combined dataset using the internal transcribed spacers (ITS), the 28S large subunit of ribosomal RNA (LSU), and partial translation elongation factor 1-alpha (tef), actin (act) and glyceraldehyde-3-phosphate dehydrogenase (gapdh) loci. Phylogenetically, these Phyllosticta isolates formed a clade in the P. concentrica species complex, and clustered with P. pilospora and P. spinarum. Morphologically, isolates in this clade differed from P. pilospora and P. spinarum by the size of conidiogenous cells and conidia, and the absence of an apical conidial appendage. As a result, these isolates were described as a novel species Phyllosticta rizhaoensis. Pathogenicity was confirmed using Koch's postulates, which showed that P. rizhaoensis could induce leaf blight symptoms on O. japonicus in China. Citation: Wang C-B, Wang T-T, Ma C-Y, Xue H, Li Y, Piao C-G, Jiang N (2023). Phyllosticta rizhaoensis sp. nov. causing leaf blight of Ophiopogon japonicus in China. Fungal Systematics and Evolution 11: 43-50. doi: 10.3114/fuse.2023.11.03.

Analysis of the mechanism of Sophorae Flavescentis Radix in the treatment of intractable itching based on network pharmacology and molecular docking.

OBJECTIVE: Sophorae Flavescentis Radix (Kuh-seng, SFR), a Traditional Chinese Medicine (TCM), is widely used alone or within a TCM formula to treat pruritus, especially histamine-independent intractable itching. In the previous study, potential antipruritic active components of the SFR were screened based on cell membrane immobilized chromatography (CMIC), revealing oxymatrine (OMT) as an antipruritic agent. However, the low oral bioavailability (OB) of OMT cannot explain the antipruritic effect of SFR when administered orally in clinic. In this study, we investigated the antipruritic effects and underlying mechanisms of orally administered SFR. MATERIALS AND METHODS: A network pharmacology and molecular docking were employed to screen the active components of SFR and predict their binding to disease-related target proteins, while the potential mechanisms were explored with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The binding energy between components and target proteins was calculated by molecular docking. RESULTS: The SFR-components-targets-intractable itching Protein-Protein Interactions (PPI) network was established, and 22 active components and 42 targets were screened. The GO enrichment analysis showed that the key target genes of SFR were related to nuclear receptors, transcription factors, and steroid hormone receptors. The results of the KEGG enrichment pathway analysis include Hepatitis B, epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor resistance, advanced glycation end product (AGE)-receptor for AGE (RAGE) signaling pathway in diabetic complications, etc. Molecular docking showed that three key target proteins in the network, the vascular endothelial growth factor A (VEGFA), epidermal growth factor receptor (EGFR) and caspase-3 (CASP3), have higher binding activities with inermine, phaseolin and kushenol O, respectively; the binding energy of each pair is stronger than that of the target protein-corresponding inhibitors. CONCLUSIONS: The complexity of the SFR-components-targets-intractable itching network demonstrated the holistic treatment effect of SFR on intractable itching. The partial coherence between results screened by CMIC in the previous study and network pharmacology demonstrated the potential of network pharmacology in active component screening. Inermine screened from both CMIC and network pharmacology is a VEGFA inhibitor, which possibly accounts for the antipruritic effect of orally administered SFR.

[Exploration on total parathyroidectomy for secondary hyperparathyroidism].

Objective: To evaluate the clinical significance of dissection parathyroidectomy for secondary hyperparathyroidism (SHPT) in patients with renal disease on maintenance dialysis. Methods: We retrospectively reviewed 195 patients with SHPT treated in the Department of Otolaryngology & Head and Neck Surgery of Beijing Civil Aviation General Hospital between September 2009 and September 2017, including 92 males and 103 females, aged from 23 to 77 years old. There were 167 patients by operated firstly and 28 patients by operated secondly for persistent or recurrent SHPT after operation. All patients received dissection parathyroidectomy with parathyroid autograft in the sternocleidomastoid. The easement of symptoms, the levels of serum intact parathyroid hormone (iPTH), serum-ionized calcium, phosphorus, and hemoglobin were compared before and after operation. Data were analyzed by SPSS 22.0 software. Results: Confirmed by postoperative pathology, a total of 804 hyperplastic parathyroid glands were removed in 195 patients with SHPT. Among them, 765 parathyroid glands were clearly identified and located with naked eye. The anatomic distribution of the glands showed 577 (75.4%) in the tracheoesophageal groove. The incidence of ectopic parathyroid glands was 24.6% (188/765). Other 39 (4.9%) hyperplastic parathyroid glands from 22(11.3%) patients, which were not identified and located with naked eye during operation, were pathologically detected in the dissected tissue specimens. Among 195 patients, 28(14.4%) showed supernumerary parathyroid glands. No serious complications occurred after operation. Within 6 months after the operation, the bone pain and skin itch symptoms were completely relieved and, also, the symptoms of muscle weakness, restless leg, anemia and poor sleep quality were significantly alleviated. Following-up at 6 months after surgery showed the serum levels of iPTH [(70.31±60.12) pg/ml], calium [(2.13±0.22) mmol/L], and phosphorus [(1.17±0.27) mmol/L] decreased significantly respectively compared with the preoperative serum levels of iPTH [(1 501.02±167.26) pg/ml], calium [(2.40±0.32) mmol/L], and phosphorus[(2.27±0.50)mmol/L], all with statistically significant differences (P<0.01); the levels of hemoglobin [(120.32±10.63) g/L] and hematocrit [(39.20±3.21)%] were higher than the preoperative levels of hemoglobin[(104.11±15.17) g/L] and hematocrit [(31.25±5.12)%], both with statistically significant differences (t valve was 12.22,18,37,respectively, all P<0.05). Conclusions: Dissection parathyroidectomy is a beneficial and safe surgical procedure for patients with medically refractory SHPT.

Mechanisms of vasorelaxation induced by total flavonoids of Euphorbia humifusa in rat aorta.

Euphorbia humifusa Willd. (EH), rich in flavonoids, has long been used for the treatment of bacillary dysentery and enteritis in China, and is known to have antioxidant, hypotensive and hypolipidemic properties. However, the vasorelaxant effect of total flavonoids of EH (TFEH) and action mechanisms are not clearly defined yet. The aim of the present study was to investigate the effects of TFEH on the vascular tension and its underlying mechanisms. Experiments were performed in rat thoracic aorta using the organ bath system. TFEH (0.01 - 100 µg/ml) caused a concentration-dependent vasorelaxation, which was dependent on a functional endothelium, and were significantly attenuated by inhibitors of endothelial NO synthase, its upstream signaling pathway, PI3K/Akt, and soluble guanylate cyclase, but not by blockade of KCa channel, KATP channel, cyclooxygenase, muscarinic and ß-adrenergic receptors. Extracellular Ca2+ depletion, and pre-treatment with modulators of the store-operated Ca2+ entry channels, Gd3+ and 2-aminoethyl diphenylborinate, significantly attenuated the TFEH-induced vasorelaxation. Our findings suggest that TFEH elicit vasorelaxation via endothelium-dependent NO-cGMP pathway through activation of PI3K/Akt- and Ca2+-eNOS-NO signaling. Further, it is suggested that TFEH-induced activation of the NO-soluble guanylate cyclase-cGMP-protein kinase G signaling relaxes vascular smooth muscle cells through an inhibition of the L-type Ca2+ channel activity.

Construction and transformation of expression vector containing Panax japonicus SS gene.

Panax japonicus C.A. Meyer, a perennial herb belonging to the Araliaceae ginseng genus, is one of the seven rare and endangered Chinese medical herbs. By cloning the SS segment, the expression vectors pCXSN-PjSS and pCXSN-antiPjSS were constructed and introduced into Agrobactria LBA4404, which is used for engineering bacteria. Polymerase chain reaction (PCR) showed that the PjSS, antiPjSS, and Hyg were integrated in Nicotiana tabacum. Reverse transcription-PCR indicated that the PjSS was transcribed into mRNA in N. tabacum and was highly expressed, while the antiPjSS was not expressed. Detection of Ginsenoside Re content showed that transgenetic N. tabacum can increase the content of Ginsenoside Re and that anti-transgenetic N. tabacum decreased the content of Ginsenoside Re.

Extracts from Plastrum testudinis promote proliferation of rat bone-marrow-derived mesenchymal stem cells.

OBJECTIVES: The purpose of this study is to identify active components of PT involved in promoting proliferation of MSCs and to investigate its mechanism. PT was extracted with petroleum ether, ethyl acetate, ethanol and water respectively. MATERIALS AND METHODS: Evidence provided by MTT, HE stain, BrdUrd, PCNA immunoreactivity and cell cycle indicated that Plastrum Testudinis Extracted with ethyl acetate (PTE) is the only active components responsible for increasing MSCs proliferation. RESULTS: This finding leads us to identify the chemical component of PTE. Steroid, fatty acids and their esters components in PTE were determined by GC-MS and HPLC. The mechanism of PTE action may be associated with the up-regulation of BMP4. CONCLUSIONS: Our findings give novel insights into the promoting effects of Plastrum Testudinis on proliferation of MSCs and help to identify the chemical component and to clarify the mechanism of its pharmacological activities.

Effects of electro-acupuncture on the expression of c-jun and c-fos in spared dorsal root ganglion and associated spinal laminae following removal of adjacent dorsal root ganglia in cats.

This study evaluated the plastic changes of c-jun and c-fos in the right sixth lumbar dorsal root ganglion (L6 DRG), Rexed's lamina II in representative spinal segments L3, L5, and L6 and in the nucleus dorsalis (ND) at L3 segments after electro-acupuncture (EA) in cats subjected to removal of L1-L5 and L7-S2 DRG. Following dorsal root ganglionectomy, there was a significant increase in the density of c-jun immunoreactivity in the neurons and glia in spinal lamina II and in the ND; there was also marked elevation in the expression of c-fos in ND. In both cases there was no change in the c-jun and c-fos immunoreactivity in the DRG. After EA in the operated animals, there was an up-regulation in the expression of c-jun in the L6 DRG and the associated spinal lamina II; however, increased c-fos expression was detected only in the L6 DRG. Western blot and RT-PCR were also performed to quantitatively explore the mRNA and protein expression changes in the spinal dorsal horn and associated DRG. Following partial deafferentation, there was a significant increase in the protein level of both c-jun and c-fos in the dorsal horn, while, in both cases there was no change in c-jun and c-fos protein and mRNA in the DRG. After EA in the operated animals, both c-jun protein and its mRNA in the L6 DRG as well as the associated dorsal horn of L6 spinal segment were upregulated, but increased c-fos protein and its mRNA was observed only in the L6 DRG. These findings suggested that c-jun and c-fos might be related to the acupuncture promoted spinal cord plasticity as reported previously.