Five New Species of Gymnopilus from Xizang Autonomous Region of China and Surrounding Areas.

The species of Gymnopilus (Hymenogastraceae, Agricales) are commonly recognized as wood-decaying fungi. Certain members of this genus have been identified as psilocybin-producing mushrooms. Gymnopilus exhibits a diverse range and has a global distribution. In this study, a total of seventy-eight specimens were gathered from ten provinces in China. A comprehensive molecular phylogenetic analysis was conducted, employing gene sequences including ITS, nrLSU, nrSSU, rpb1, rpb2, and tef1-α. Additionally, morphological examinations were also carried out. The phylogenetic topology of Gymnopilus from this study generally agreed with previous studies and facilitated the identification of all those specimens. As a result, eleven species, including five newly discovered ones named Gy. gyirongensis, Gy. variisporus, Gy. tomentosiceps, Gy. tenuibasidialis, and Gy. aurantipileatus, were recognized. Significantly, four of the five newly identified species are native to the Xizang Autonomous Region, emphasizing their specialization in this distinctive habitat. This research contributes to our comprehension of Gymnopilus diversity and lays the groundwork for the conservation and sustainable utilization of Gymnopilus resources.

Erratum: [Corrigendum] Dl­3­n­butylphthalide alleviates cognitive impairment in amyloid precursor protein/presenilin 1 transgenic mice by regulating the striatal­enriched protein tyrosine phosphatase/ERK/cAMP­response element­binding pro­â€‹tein signaling pathway.

[This corrects the article DOI: 10.3892/etm.2022.11248.].

Emodin Ameliorates High Glucose-Induced Podocyte Apoptosis via Regulating AMPK/mTOR-Mediated Autophagy Signaling Pathway.

OBJECTIVE: To investigate the effect of emodin on high glucose (HG)-induced podocyte apoptosis and whether the potential anti-apoptotic mechanism of emodin is related to induction of adenosine-monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)-mediated autophagy in podocytes (MPC5 cells) in vitro. METHODS: MPC5 cells were treated with different concentrations of HG (2.5, 5, 10, 20, 40, 80 and 160 mmol/L), emodin (2, 4, 8 µ mol/L), or HG (40 mmol/L) and emodin (4 µ mol/L) with or without rapamycin (Rap, 100 nmol/L) and compound C (10 µ mol/L). The viability and apoptosis of MPC5 cells were detected using cell counting kit-8 (CCK-8) assay and flow cytometry analysis, respectively. The expression levels of cleaved caspase-3, autophagy marker light chain 3 (LC3) I/II, and AMPK/mTOR signaling pathway-related proteins were determined by Western blot. The changes of morphology and RFP-LC3 fluorescence were observed under microscopy. RESULTS: HG at 20, 40, 80 and 160 mmol/L dose-dependently induced cell apoptosis in MPC5 cells, whereas emodin (4 µ mol/L) significantly ameliorated HG-induced cell apoptosis and caspase-3 cleavage (P<0.01). Emodin (4 µ mol/L) significantly increased LC3-II protein expression levels and induced RFP-LC3-containing punctate structures in MPC5 cells (P<0.01). Furthermore, the protective effects of emodin were mimicked by rapamycin (100 nmol/L). Moreover, emodin increased the phosphorylation of AMPK and suppressed the phosphorylation of mTOR. The AMPK inhibitor compound C (10 µ mol/L) reversed emodin-induced autophagy activation. CONCLUSION: Emodin ameliorated HG-induced apoptosis of MPC5 cells in vitro that involved induction of autophagy through the AMPK/mTOR signaling pathway, which might provide a potential therapeutic option for diabetic nephropathy.

Dl-3-n-butylphthalide alleviates cognitive impairment in amyloid precursor protein/presenilin 1 transgenic mice by regulating the striatal-enriched protein tyrosine phosphatase/ERK/cAMP-response element-binding protein signaling pathway.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive impairment and the deposition of amyloid plaques in the brain. In a transgenic mouse model of AD, cognitive impairment and synaptic dysfunction were revealed to be associated with soluble amyloid oligomers and to occur prior to plaque formation. The results of our previous studies revealed that striatal-enriched protein tyrosine phosphatase (STEP)61 negatively regulated the ß-amyloid protein-mediated ERK/cAMP-response element-binding protein (CREB) signaling pathway. Dl-3-n-butylphthalide (NBP) is a synthetic compound approved by the Food and Drug Administration of China for the treatment of ischemic stroke in 2002. Studies have shown that the neuroprotective effects of NBP involve multiple mechanisms. The present study further explored the mechanism of NBP therapy in amyloid precursor protein (APP)/presenilin 1 (PS1) transgenic mice, and the involvement of the STEP/ERK/CREB signaling pathway. The results suggested that NBP treatment effectively ameliorated the spatial learning and memory impairment of the APP/PS1 transgenic mice, which was assessed using a Morris water maze. In addition, NBP reduced amyloid-induced activation of STEP61 levels, while increasing phosphorylated (p)-ERK1/2 and p-CREB levels in the cerebral cortex and hippocampus of APP/PS1 transgenic mice by western blotting and immunostaining. In conclusion, the present study provided evidence to suggest that the new drug NBP improved amyloid-induced learning and memory deficits, likely through the regulation of the STEP/ERK/CREB pathway. The results revealed that NBP, as a multi-target drug, may exert a neuroprotective effect. Therefore, NBP may serve as an effective treatment for AD.

A long-lasting biological larvicide against the dengue vector mosquito Aedes albopictus.

BACKGROUND: Aedes albopictus is the primary vector of mosquito-borne diseases, including dengue and chikungunya, in China. The management of vector mosquitoes is the primary strategy for the control of such infectious diseases. The gravid Ae. albopictus prefers to skip-oviposit its eggs into different small water containers, and the management of these breeding places is critical for mosquito control. Bacillus thuringiensis subspecies Israelensis (Bti) is a useful biological larvicide, but the effective period of the currently available commercial product is relatively short. This study aimed to develop a long-lasting formulation of Bti to control the dengue vector mosquito Ae. albopictus. RESULTS: Water-soluble polyethylene glycols and water-insoluble hexadecanol were mixed with Bti to develop the long-lasting formulation Bti-BLOCK, based on the solid dispersion technique. The controlled release of Bti-BLOCK and its effect on Ae. albopictus were assayed in the laboratory and in the field. The results showed that Bti toxins were slowly released from Bti-BLOCK into the water and maintained at an effective dose for at least 6 months. Bti-BLOCK caused high mortality during the immature stage (>90%) and achieved full inhibition during pupation (100%). The efficacy lasted at least 12 weeks in the laboratory and 6 weeks in the field. Furthermore, we confirmed an 89% reduction in Ae. albopictus density and a reduction in the R0 of dengue to a low-risk level after 6 months of open-field interventions. CONCLUSIONS: We developed a long-lasting biological larvicide, Bti-BLOCK, which displayed very good efficacy in the control of the dengue vector mosquito Ae. albopictus.

Delayed Facial Palsy After Microvascular Decompression for Hemifacial Spasm.

OBJECTIVE: We explored the risk factors for the occurrence of delayed facial paralysis (DFP) after microvascular decompression (MVD) for hemifacial spasm (HFS). METHODS: A retrospective study was conducted of 636 patients who had undergone MVD for HFS by the same neurosurgery department of China-Japan Friendship Hospital from January 2006 to May 2016. Of the 636 patients, 50 (7.9%) had presented with DFP, which had developed from 2 to 60 days postoperatively (average, 12.9 ± 10.0005 days). All 50 patients with DFP had recovered completely within 10-300 days (average, 88.7 ± 61.389 days) after the onset of DFP. We randomly selected 100 patients from the 586 patients without DFP as the control group. Univariate and multivariate logistic analyses were used to analyze the risk factors involved in the occurrence of DFP. RESULTS: Univariate analysis showed that the disease course was the only factor associated with the development of DFP (P = 0.003). Furthermore, on multivariate logistic analysis, the course of HFS was the only risk factor associated with the development of DFP (P = 0.01). Additionally, the Spearman test revealed a positive correlation between the onset of DFP and the duration of the DFP symptoms (rs = 0.682; P < 0.001). CONCLUSION: Although DFP frequently occurred after MVD, it can recover spontaneously. The longer the course of HFS, the more frequently DFP will occur after MVD. The earlier that DFP develops, the shorter will be the time to recovery.

Preservation or Ligation of Splenic Vessels During Spleen-Preserving Distal Pancreatectomy: A Meta-Analysis.

Purpose/aim: Spleen preservation distal pancreatectomy (SPDP) can be achieved by either splenic vessel preservation distal pancreatectomy (SVP-DP) or Warshaw technique (WT). Although studies comparing SVP-DP with WT have been reported, controversies exist. The aim of our study is to assess and compare the safety and feasibility of SVP-DP and WT. Materials and methods: Two authors searched the online database independently till April 30, 2017. Data extraction and quality assessment were performed independently by two authors. Short- and long-term outcomes of WT and SVP-DP were evaluated. Subgroup analysis was performed on laparoscopic surgery. Odds ratios (OR) with 95% confidence interval (CI) and mean difference (MD) with 95% CI were estimated. Results: A total of 664 patients from 11 retrospective cohort studies were included. Meta-analysis showed the WT group had a significantly higher incidence of splenic infarction (OR = 0.12; 95% CI: 0.07-0.20; p < 0.00001) and gastric/epigastric varices (OR = 0.11; 95% CI: 0.05-0.24; p < 0.00001). And more patients suffering from splenic infarction from WT group needed further splenectomy (OR = 0.13; 95% CI: 0.02-0.84; p = 0.03). While there was no difference between the two procedures in terms of pancreatic fistula (OR = 0.55; 95% CI: 0.25-1.19; p = 0.13), overall morbidity (OR = 0.87; 95% CI: 0.59-1.30; p = 0.50) and hospital stay (MD = -0.45; 95% CI: -1.73-0.82; p = 0.49). Conclusions: Due to relatively higher risk of postoperative splenic infarction, gastric/epigastric varices and Clavien-Dindo III-V complications, WT is not as safe as SVP-DP. However, well-conducted randomized clinical trials are still needed due to the limitations of current studies.

Construction of an efficient genomic editing system with CRISPR/Cas9 in the vector mosquito Aedes albopictus.

Aedes (Stegomyia) albopictus, also known as the Asian tiger mosquito, is a mosquito which originated in Asia. In recent years, it has become increasingly rampant throughout the world. This mosquito can transmit several arboviruses, including dengue, Zika and chikungunya viruses, and is considered a public health threat. Despite the urgent need of genome engineering to analyze specific gene functions, progress in genetical manipulation of Ae. albopictus has been slow due to a lack of efficient methods and genetic markers. In the present study, we established targeted disruptions in two genes, kynurenine hydroxylase (kh) and dopachrome conversion enzyme (yellow), to analyze the feasibility of generating visible phenotypes with genome editing by the clustered regularly interspaced short palindromic repeats (CRISPR) / CRISPR-associated protein 9 (Cas9) system in Ae. albopictus. Following Cas9 single guide RNA ribonucleoprotein injection into the posterior end of pre-blastoderm embryos, 30%-50% of fertile survivors produced alleles that failed to complement existing kh and yellow mutations. Complete eye and body pigmentation defects were readily observed in G1 pupae and adults, indicating successful generation of highly heritable mutations. We conclude that the CRISPR/Cas9-mediated gene editing system can be used in Ae. albopictus and that it can be adopted as an efficient tool for genome-scale analysis and biological study.

Danhong Injection Alleviates Mechanical Allodynia via Inhibiting ERK1/2 Activation and Elevates BDNF Level in Sciatic Nerve in Diabetic Rat.

Danhong injection (DHI) has been widely used in China for cardiocerebrovascular diseases treatments. And in this study, we demonstrated the therapeutic effect of DHI on experimental diabetic neuropathy for the first time. Methods. Streptozotocin- (STZ-) induced SD rats were used. In experiment 1, 4-week treatment with DHI or saline started 4 weeks after STZ injection; mechanical allodynia was measured before and every 2 weeks after STZ injection. In experiment 2, chronic intrathecal infusion of U0126 was conducted during the 8th week of diabetes. Phosphorylated and total ERK1/2 in spinal cord were analyzed by western blot. BDNF level in sciatic nerve was evaluated by ELISA. Results. DHI treatment significantly alleviated mechanical allodynia at the end of the study and downregulated the expression of phosphorylated ERK1/2 in spinal cord. In addition, DHI treatment also elevated brain-derived neurotrophic factor (BDNF) level in sciatic nerve of DPN rat. In experiment 2, inhibition of ERK1/2 activation was confirmed to result in the alleviation of mechanical allodynia. Conclusions. We demonstrated that DHI was able to alleviate mechanical allodynia in diabetic neuropathy rat through inhibiting the activation of ERK1/2. The reduction of BDNF content in sciatic nerve was also partially reversed by DHI treatment.

Two-Point Discrimination Predicts Pain Relief after Lower Limb Nerve Decompression for Painful Diabetic Peripheral Neuropathy.

BACKGROUND: Peripheral nerve decompression surgery has been reported to be effective for pain reduction in patients with painful diabetic peripheral neuropathy. The aim of this study was to characterize which patients may have more pain relief benefits in the lower limbs after nerve decompression surgery. METHODS: A retrospective study was conducted. Pain levels were measured with the Numerical Rating Scale. Treatment effects were classified by either substantial relief (at least 50 percent reduction in Numerical Rating Scale score compared with preoperative Numerical Rating Scale score) or nonsubstantial relief (<50 percent reduction or worse in Numerical Rating Scale score) at 12 months based on established criteria. Sex, age, body mass index, duration of diabetes mellitus, duration of diabetic peripheral neuropathy pain, preoperative Numerical Rating Scale score, and two-point discrimination were evaluated using univariate and logistic regression analysis. RESULTS: The mean preoperative Numerical Rating Scale score (8.65 ± 1.29) decreased significantly 6 days (3.56 ± 2.22; p < 0.01), 6 months (3.03 ± 2.11; p < 0.01), and 12 months (3.44 ± 2.36; p < 0.01) after surgery; 64.7 percent of patients had substantial pain relief at 12 months. According to univariate and logistic regression analysis, better two-point discrimination was associated with substantial pain relief (OR, 3.700; p = 0.046, logistic regression analysis). CONCLUSIONS: Nerve decompression surgery was able to alleviate pain in patients with painful diabetic peripheral neuropathy. Two-point discrimination may be a predictive factor for the prognosis of painful diabetic peripheral neuropathy after nerve decompression surgery. CLINICAL QUESTION/LEVEL OF EVIDENCE: Risk, III.

Tanshinone IIA Improves Painful Diabetic Neuropathy by Suppressing the Expression and Activity of Voltage-Gated Sodium Channel in Rat Dorsal Root Ganglia.

Painful diabetic neuropathy (PDN) is one of the intractable complications of diabetes mellitus, which manifest as exaggerated pain perception. Previous studies showed that Tanshinone IIA (TIIA), one of the major bioactive extracts of Salvia miltiorrhiza Bunge, have obvious analgesic effect on different types of pain process, and the underlying analgesic mechanisms are not fully understood. The present study combined the behavioral, electrophysiological and biochemical methods to elucidate the analgesic mechanism of TIIA, using streptozotocin (STZ)-induced PDN rat models. Intraperitoneal injection (i.p.) of TIIA for 3 weeks in PDN rats significantly improved mechanical allodynia and thermal hyperalgesia. Patch clamp recordings showed that the excitability of dorsal root ganglion (DRG) nociceptive neuron was increased in diabetic state, and TIIA treatment effectively recovered the subnormality, which was achieved by preventing augments of both Tetrodotoxin-sensitive (TTX-resistant) and Tetrodotoxin-sensitive (TTX-S) sodium currents. Further, the protein expressions of voltage-gated sodium channels (VGSCs) α-subunits Nav1.3, Nav1.7 and Nav1.9 increased in DRG of diabetic rats and were normalized by TIIA application. In conclusion, this study provides evidence that the TIIA attenuated PDN by effecting VGSCs activities and expressions, indicating that the TIIA could be a promising agent for PDN treatment.

Phytochemical and Pharmacological Studies on the Genus Psoralea: A Mini Review.

The genus Psoralea, which belongs to the family Fabaceae, comprises ca. 130 species distributed all over the world, and some of the plants are used as folk medicine to treat various diseases. Psoralea corylifolia is a typical example, whose seeds have been widely used in many traditional Chinese medicine formulas for the treatment of various diseases such as leucoderma and other skin diseases, cardiovascular diseases, nephritis, osteoporosis, and cancer. So, the chemical and pharmacological studies on this genus were performed in the past decades. Here, we give a mini review on this genus about its phytochemical and pharmacological studies from 1910 to 2015.

[Effect of Tongmai Jiangtang Capsule on Experimental Diabetic Peripheral Neuropathy Rats].

OBJECTIVE: To observe the effect of Tongmai Jiangtang Capsule (TJC) on experimental diabetic peripheral neuropathy (DPN) rats. METHODS: Forty Wistar rats were divided into the TJC group, the mecobalamin treatment group, the model group, and the normal group according to random digit table, 10 in each group. Except rats in the normal group, DPN rat model was prepared using intraperitoneally in- jecting streptozotocin (STZ) in the rest rats. One rat in the model group died during the modeling. Different drugs were administered by gastrogavage to rats in corresponding groups from the 8th week after successful modeling. TJC (0.23 g crude drugs/mL, 10 mL/kg) was administered to rats in the TJC group by gastrogavage. Suspension of mecobalamin and normal saline (10 mL/kg, 0.05 mg/mL) was administered by gastrogavage to rats in the mecobalamin treatment group to the end of the 12th week. Meanwhile, equal volume of distilled water was administered by gastrogavage to rats in the model group and the normal group. Peripheral nerve conduction velocity was detected in each group. Gait analysis was performed. Changes of intraepidermal nerve fiber were observed by immunohistochemical assay. Pathological changes of tibial nerve tissue were observed using HE staining. RESULTS: (1) Compared with the normal group, the nerve conduction velocity was slowed down; print length (PL), intermediary toe spread (ITS), and toe spread (TS) were added in the model group, with statistical difference (P <0. 01). Compared with the mod- el group, nerve conduction velocity was speeded; PL and ITS decreased in the TJC group and the mecobal- amin treatment group, with statistical difference (P <0. 01). Besides, the nerve conduction velocity was superior in the TJC group than in the mecobalamin treatment group, with statistical difference (P <0. 05). (2) Immunohistochemical results showed, the staining of intraepidermal nerve fiber was not clear and dispersedly distributed in the model group, with no nerve fiber staining in local regions. Nerve fibers were not regular in lesser amount and shallow stained in the mecobalamin treatment group, with no nerve fiber staining in local regions. Nerve fibers were not regular in lesser amount and dispersedly distributed in the TJC group. (3) HE staining showed that tibial nerve tissue was severely swollen with swollen myelin sheath in the mod- el group. It was difficult to identity myelin sheath. Vaculole degenerated in local regions. Swollen axon could be seen. Partial axons were separated and degenerated. In the mecobalamin treatment group tibial nerve tissue was edematous with swollen myelin sheath. It was difficult to identity myelin sheath. Axons were locally separated. In the JMC group tibial nerve tissue was swollen with unclear myelin sheath and swollen axons. CONCLUSION: TJC could improve peripheral neuropathy of diabetic rats.

[Complications of selective posterior rhizotomy for lower limb spasticity of cerebral palsy].

OBJECTIVE: To investigate the complications of spastic cerebral palsy with selective posterior rhizotomy (SPR). METHODS: In the study, 2 593 patients who had undergone SPR from January 2000 to September 2012 were followed-up for at least one year. The complications were classified. RESULTS: Peri-operative complications: pulmonary system complications including bronchial spasm (5 cases, 0.19%) and aspiration pneumonia (4 cases, 0.15%); digestive system complications including abdominal bloating (145 cases, 5.6%) and colic (80 cases, 3.1%); urinary system complications including temporary bladder dysfunction (54 cases, 2.1%) and urinary tract infection (6 cases, 0.23%); peripheral nervous system complications including lower extremity weakness (327 cases, 12.6%) and lower extremity sensory disturbances (140 cases, 5.4%); central nervous system complications including headache (112 cases, 4.3%) and epileptic seizures (4 cases, 0.15%). None spinal or intracranial infection, intraspinal hematoma or intracranial hemorrhage were identified. General surgery complications including back pain (1 382 cases, 53.3%), delay wound healing caused by infection (5 cases, 0.19%) and cerebrospinal fluid leakage (8 cases, 0.31%). Long-term follow-up complications including lower limb decreased exercise capacity (incidence: 7.33%) and lower extremity sensory disturbance (incidence: 5.59%). Urination occurred in only one case and defecation function disturbance with no sexual dysfunction was identified. The incidences of scoliosis, thoracic kyphosis, spondylolisthesis and long-term back pain were 7.23% (31/429), 4.2% (18/429), 10.49% (45/429) and 9.72% respectively. CONCLUSION: SPR is one of the effective and safe surgical treatments for spastic cerebral palsy. Valid methods should be applied to reduce the incidence of postoperative complications, such as choosing the appropriate patients, meticulously operating in the surgery, assistance of electrophysiological guidance, reinforcing perioperative management and regular rehabilitation training after operation.

[Chemical constituents from the fruits of Areca catechu].

OBJECTIVE: To investigate the chemical constituents of the fruits of Areca catechu. METHODS: The chemical constituents were isolated by silica gel, RP-18 and Sephadex LH-20 column chromatographies. Their structures were determined by chemical and spectroscopic methods. RESULTS: Thirteen compounds were isolated from the 95% ethanol extract of the fruits of Areca catechu. Their structures were identified as isorhamnetin (1), quercetin (2), liquiritigenin (3), 5,7,4'-trihydroxy-3',5'-dimethoxyflavanone (4), (+)-catechin (5), resveratrol (6), ferulic acid (7), vanillic acid (8), 5,8-epidioxiergosta-6,22-dien-3beta-ol (9), stigmasta-4-en-3-one (10), beta-sitosterol (11), cycloartenol (12), and de-O-methyllasiodiplodin (13), respectively. CONCLUSION: Compounds 2-4,6,7,9,10, 12,13 are isolated from this plant for the first time.

Immunofluorescence laser confocal expression and localization study of rat nerve growth guidance cues Netrin-1 and Slit2 after spinal cord injury.

OBJECTIVE: To observe the expression and distribution of adult rat axon guidance cues Netrin-1 and Slit2 at different time points after spinal cord injury and to investigate the guidance mechanism of regenerated axons. METHODS: Twenty adult Sprague Dawley (SD) rats were divided randomly into five groups with 4 in each. Four groups of them were used to make Allen's spinal cord punch models and we took materials randomly from one of them on the 2nd, 4th, 7th and 14th day respectively after operation. The left one group was taken as the control group. Immunofluorescence laser confocal scan was used to examine the co-expression and localization of Netrin-1 and Slit2 proteins in the injured site of the spinal cord. RESULTS: Within two weeks after SCI, the expression of Netrin-1 and Slit2 proteins increased temporarily and there was co-expression of them on the neuron plasma membrane. CONCLUSIONS: Synchronous high expression and co-expression of axon attractant Netrin-1 and repellent Slit2 are found in the adult rat injured spinal cord in the damaged local and vicinity parts, and probably, they act as the key regulators of axon guidance regeneration.

A novel C2-domain phospholipid-binding protein, OsPBP1, is required for pollen fertility in rice.

Pollen fertility is a crucial factor for successful pollination and essential for seed formation. Recent studies have suggested that a diverse range of internal and external factors, signaling components and their related pathways are likely involved in pollen fertility. Here, we report a single C2-domain containing protein, OsPBP1, initially identified through cDNA microarray analysis. OsPBP1 is a single copy gene and preferentially expressed in pistil and pollen but down-regulated by pollination. OsPBP1 had a calcium concentration-dependent phospholipid-binding activity and was localized mainly in cytoplasm and nucleus, but translocated onto the plasma membrane in response to an intracellular Ca(2+) increase. Pollen grains of antisense OsPBP1 transgenic lines were largely nonviable, germinated poorly in vitro and of low fertility. OsPBP1 protein was localized in a region peripheral to pollen wall and vesicles of elongating pollen tube, and its repressed expression reduced substantially this association and led to alteration of microfilament polymerization during pollen germination. Taken together, these results indicate that OsPBP1 is a novel functional C2-domain phospholipids-binding protein that is required for pollen fertility likely by regulating Ca(2+) and phospholipid signaling pathways.