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During quantitative real-time polymerase chain reaction (RT-qPCR) data analysis, the selection of optimal housekeeping gene is necessary to ensure the accuracy of results. It is noteworthy that housekeeping genes commonly used in adult studies may not be applicable for fetus. However, the stability analysis of housekeeping gene in fetal kidney has not been reported. This study intends to screen the applicable compound housekeeping genes in rat fetal kidney. In this study, eight housekeeping genes used in kidney studies based on literature reports (GAPDH, ACTB, 18S, HPRT, YWHAZ, HMBS, PPIA, and TBP) were selected as the research object. Their expression levels in the rat fetal kidney in physiological condition and the intrauterine growth retardation (IUGR) model induced by prenatal dexamethasone exposure (PDE) (0.2 mg/kg·day from gestation Days 9 to 20) was measured. Furthermore, these eight housekeeping genes were used to conduct relative quantitative analysis of nephrin expression in the fetal kidney in PDE-induced IUGR model, to compare the influence of choosing different housekeeping gene on data analysis of nephrin expression and to verify the reliability of selected compound housekeeping genes. In this study, stable housekeeping genes of fetal kidney tissues in PDE-induced IUGR model were identified: ACTB, GAPDH, TBP, and HMBS for males; ACTB, YWHAZ, and GAPDH for females. Besides, our results suggest that ACTB + GAPDH were the best compound housekeeping genes for normalization analysis in male fetal kidney studies, and ACTB + YWHAZ in females. This study will provide an experimental evidence basis for the selection of housekeeping genes in the RT-qPCR experiment in renal development toxicology-related models.

Alhagi pseudalhagi, commonly known as camel thorn, is used as an indigenous medicinal plant in China. The present study was designed to elucidate the structure of a novel polysaccharide, APP90-2, isolated from Alhagi pseudalhagi and evaluate its osteogenic activity. A homogeneous polysaccharide (APP90-2) was obtained from A. pseudalhagi via DEAE-52 and Sephacryl S-100 columns, with a molecular weight of 5.9 kDa. Monosaccharide, GC–MS, and NMR analyses showed that APP90-2 consisted of α- l -Rhap-(1→, →3)-α- l -Araf-(1→, →5)-α- l -Araf-(1→, →4)-β- d -Xylp-(1→, α- d -Glcp-(1→, →3,5)-α- l -Araf-(1→, →4)-β- d -GlcAp-(1→, →4)-3-OAc-α- d -Glcp-(1→, →3)-α- d -Galp-(1→, →3)-β- d -GalAp-(1→, →4)-α- d -Galp-(1→, →6)-α- d -Manp-(1→, →4,6)-β- d -Galp-(1→, and →3,6)-β- d -Glcp-(1→ with relative molar ratios of 4.1:1.8:6.1:6.7:1.7:1.0:1.5:2.7:2.4:1.1:2.3:2.6:1.4:2.0. Morphological analyses revealed that APP90-2 interacted with Congo-red and had an obvious honeycomb structure. Additionally, APP90-2 significantly promoted proliferation, differentiation, and mineralization of MC3T3-E1 cells, indicating that APP90-2 exhibited pronounced osteogenic activity. Therefore, our findings suggest that A. pseudalhagi may be used as an alternative medicine or health supplement for the prevention and treatment of osteoporosis.

T-006, a small-molecule compound derived from tetramethylpyrazine (TMP), has potential for the treatment of neurological diseases. In order to investigate the effect of T-006 prophylactic treatment on an Alzheimer's disease (AD) model and identify the target of T-006, we intragastrically administered T-006 (3 mg/kg) to Alzheimer's disease (AD) transgenic mice (APP/PS1-2xTg and APP/PS1/Tau-3xTg) for 6 and 8 months, respectively. T-006 improved cognitive ability after long-term administration in two AD mouse models and targeted mitochondrial-related protein alpha-F1-ATP synthase (ATP5A). T-006 significantly reduced the expression of phosphorylated-tau, total tau, and APP while increasing the expression of synapse-associated proteins in 3xTg mice. In addition, T-006 modulated the JNK and mTOR-ULK1 pathways to reduce both p-tau and total tau levels. Our data suggested that T-006 mitigated cognitive decline primarily by reducing the p-tau and total tau levels in 3xTg mice, supporting further investigation into its development as a candidate drug for AD treatment.

T-006, a new derivative of tetramethylpyrazine, has been recently found to protect against 6-hydroxydopamine (6-OHDA)-induced neuronal damage and clear α-synuclein (α-syn) by enhancing proteasome activity in an α-syn transgenic Parkinson's disease (PD) model. The effect of T-006 on the 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced PD model, however, has not been tested and T-006's neuroprotective mechanisms have not been fully elucidated. In this study, we further investigated the neuroprotective and neurogenic effects of T-006 and explored its underlying mechanism of action in both cellular and animal PD models. T-006 was able to improve locomotor behavior, increase survival of nigra dopaminergic neurons and boost striatal dopamine levels in both MPTP- and 6-OHDA-induced animals. T-006 treatment restored the altered expressions of myocyte enhancer factor 2D (MEF2D), peroxisome proliferator-activated receptor γ (PPARγ) co-activator 1α (PGC1α) and NF-E2-related factor 1/2 (Nrf1/2) via modulation of Akt/GSK3β signaling. T-006 stimulated MEF2, PGC1α and Nrf2 transcriptional activities, inducing Nrf2 nuclear localization. Interestingly, T-006 promoted endogenous adult neurogenesis toward a dopaminergic phenotype by activating brain-derived neurotrophic factor (BDNF) and cAMP responsive element-binding protein (CREB) in 6-OHDA rats. Our work demonstrated that T-006 is a potent neuroprotective and neuroregenerative agent that may have therapeutic potential in the treatment of PD.

PTP70-2, a novel polysaccharide isolated from Polygala tenuifolia in our previous publication, exhibits potential anti-inflammatory effects. Here, we investigate the mechanisms underlying these effects and the neuroprotective activity of PTP70-2 in lipopolysaccharide (LPS)-damaged BV2 microglial cells and neuroinflammation-injured primary cortical neurons. The results suggest that PTP70-2 dramatically reduces the LPS-stimulated inflammatory cytokines overexpression, as well as down-regulates the levels of TLR4-, MyD88-, and NF-κB-related proteins. The effect of PTP70-2 in down-regulation of proinflammatory cytokines and downstream proteins implicated in MyD88 and NF-κB signaling is related to the TLR4 pathway. Furthermore, this effect is enhanced by the co-incubation of BV2 cells with PTP70-2 and TAK242, a TLR4 inhibitor, before exposure to LPS. Importantly, PTP70-2 prevents neuroinflammation-induced neurotoxicity by mitigating ROS overproduction and MMP dissipation. Overall, the PTP70-2's anti-neuroinflammation and neuroprotection are involved to the modulation of the TLR4-mediated MyD88/NF-κB signaling pathway.

Achyranthes bidentata is a species of flowering plant that is mainly distributed in China. The A. bidentata rhizome is a famous traditional Chinese medicine that has been widely used to treat lumbago, arthritis, and bone hyperplasia. In this work, A. bidentata rhizome was isolated and purified to obtain a pectic polysaccharide (ABPB-4). Chemical and spectral analyses showed that ABPB-4 had a main chain of →4)-α-d-GalpA-(1→ and →2,4)-α-l-Rhap-(1→, and the branch chains included →4)-β-d-Galp-(1→, →6)-β-d-Galp-(1→, →3,6)-β-d-Galp-(1→, →5)-α-l-Araf-(1→ and →3,5)-α-l-Araf-(1→, and it was terminated with α-l-Araf-(1→ and β-d-Galp-(1→. At concentrations of 0.01, 0.02, and 0.04 μmol/L, ABPB-4 significantly promotes the proliferation, differentiation, and mineralization of MC3T3-E1 cells in vitro, and it appreciably enhances the mRNA expression levels of osteogenic-related genes in these cells. Overall, the results reported herein indicate that ABPB-4 has outstanding osteogenic activity, and that it may be used as an anti-osteoporosis agent in the future.

Achyranthes bidentata is a traditional Chinese medicine used to treat osteoporosis. AB90, a crude saccharide from A. bidentata, showed excellent osteoprotective effects in ovariectomized rats, and ABW90-1, an oligosaccharide purified from AB90, stimulated significant differentiation of osteoblasts. However, the osteogenic effects and underlying mechanisms of ABW90-1 have remained unknown. In the present study, we found that ABW90-1 significantly promoted ALP activity, mineralization, and the expression of osteogenic markers in MC3T3-E1 cells. ABW90-1 showed strong binding with the WNT signaling complex and BMP2 based on number of interactions, hydrogen bond length, and binding energy in silico. ABW90-1 significantly increased the expression of active-β-catenin, p-GSK-3β, LEF-1, BMP2, and p-SMAD1. Importantly, the osteogenic effects of ABW90-1 were partially suppressed by DKK-1 and Noggin, which are specific inhibitors of the WNT and BMP signaling pathways, respectively. Collectively, these findings suggest that ABW90-1 has osteogenic effects through crosstalk between WNT/β-catenin and BMP2/SMAD1 signaling pathways.

This study was aimed to investigate the effect of prenatal ethanol exposure (PEE) on the susceptibility of offspring rats to glomerulosclerosis and to explore the mechanism. Pregnant Wistar rats were intragastrically administered ethanol (4g/kg·d) from gestational day (GD) 9 to GD 20, and the control group was given equal volume of normal saline. The offspring rats were all fed with high-fat diet after weaning, and were sacrificed at postnatal week 24 (PW24). The results revealed that the adult offspring kidneys in the male and female PEE groups exhibited higher glomerulosclerosis index and interstitial fibrosis index compared with the high-fat diet control groups, accompanied by elevated serum creatinine level. The protein expression of Nephrin and WT1, which were the marker genes of podocytes, was significantly decreased, whereas the protein expression of desmin and α-SMA, the marker genes of mesenchymal cells, was remarked enhanced in the male and female PEE groups. Compared with the high-fat diet control groups, the mRNA and protein expressions of renal angiotensin II receptor type 2 (AT2R) were decreased in the male PEE group, but increased in the female PEE group. PEE increased the mRNA and protein expressions of glucocorticoid (GC) activation system and inhibited the expression of insulin-like growth factor 1 (IGF1) signaling pathway in male offspring kidney; on the contrary, in female offspring kidney, PEE inhibited the mRNA and protein expression of glucocorticoid activation system and increased the expression of IGF1 signaling pathway. Taken together, PEE increased the susceptibility of the adult offspring to glomerulosclerosis, and the programming of renal AT2R or GC-IGF1 is respectively involved in the toxicity of PEE to the male or female offspring.

This study aimed to verify the toxic effects of prenatal caffeine exposure (PCE) on the podocyte development in male offspring, and to explore the underlying intrauterine programming mechanisms. The pregnant rats were administered with caffeine (30 to 120 mg/kg⋅d) during gestational day (GD) 9 to 20. The male fetus on GD20 and the offspring at postnatal week (PW) 6 and PW28 were sacrificed. The results indicated that PCE caused ultrastructural abnormalities on podocyte, and inhibited the expression of podocyte marker genes such as Nephrin, Wilms tumor 1 (WT1), the histone 3 lysine 9 acetylation (H3K9ac) level in the Kruppel-like factor 4 (KLF4) promoter and its expression in the male offspring from GD20 to PW28. Meanwhile, the expression of glucocorticoid receptor (GR) and histone deacetylase 7 (HDAC7) in the fetus were increased by PCE. In vitro, corticosterone increased GR and HDAC7 whereas reduced the H3K9ac level of KLF4 and KLF4/Nephrin expression. KLF4 over-expression reversed the reduction of Nephrin expression, knockdown of HDAC7 and GR antagonist RU486 partially reversed the inhibitory effects of corticosterone on H3K9ac level and KLF4 expression. In conclusion, PCE caused podocyte developmental toxicity in male offspring, which was associated with corticosterone-induced low-functional programming of KLF4 through GR/HDAC7/H3K9ac pathway.

Neuronal death is among the deleterious pathological changes that occur after cerebral ischemia and can lead to transient or permanent neurological deficits. The tetramethylpyrazine analog T-006 has been shown to be a multifunctional neuroprotective agent; however, its neuroprotective effect and mechanism of action have not been studied in ischemic stroke model rats. This study investigated the neuroprotective effects of T-006 in rat stroke model using a battery of behavioral and molecular biological tests. Results indicated that T-006 treatment significantly improved neurological function and behavior. Double immunofluorescence staining showed that T-006 visibly improved the number of NeuN/BrdU, Nestin/BrdU, and DCX/BrdU cells and induced neuronal regeneration. Western blot analyses indicated that T-006 upregulated neurogenesis-related protein expression of postsynaptic density protein 95, brain-derived neurotrophic factor, synaptophysin, and myelin basic protein. Collectively, these data suggest that T-006 stimulated neurogenesis in rats with middle cerebral artery occlusion and restored neurological functions.

Polygala tenuifolia Willd. is a vital industrial-export plant in Southeast Asia, particularly China. The rhizomes of P. tenuifolia are known for their nootropic and cognition-improving effects. As such, they have been widely used in traditional Chinese medicine (TCM) for centuries. However, the active constituents responsible for the biological benefits of P. tenuifolia remain unclear. In this study, P. tenuifolia rhizomes were extracted with hot water and then isolated and purified using diethylaminoethyl cellulose (DEAE-C) 52 and Sephacryl S-100 chromatographic columns. A homogeneous heteropolysaccharide (PTP70-2) with a relative molecular weight (MW) of 65.2 kDa was obtained. Based on monosaccharide composition, Fourier-transform infrared spectroscopy (FTIR), gas chromatography–mass spectrometry (GC/MS), and nuclear magnetic resonance (NMR) analyses, PTP70-2 was composed of 11 residues: →5)-α-L-Araf-(1→, →2,5)-α-L-Araf-(1→, →3,5)-α-L-Araf-(1→, →6)-β-D-Galp-(1→, →6)-α-L-Manp-(1→, →4)-β-D-Galp-(1→, →4,6)-β-D-Galp-(1→, α-L-Araf, α-L-Rhap, α-D-GlcAp, and α-L-Manp, at a molar ratio of 7.3:1.4:5.2:15.0:2.1:2.4:5.6:8.2:1.3:1.5:1.3. Scanning electron microscopy (SEM) results showed that PTP70-2 had an irregular surface with scattered strips. Pharmacological assessments showed that PTP70-2 inhibited the production of nitric oxide (NO) in lipopolysaccharide (LPS)–induced pro-inflammatory BV2 microglial cells, and the inhibitory effect of 3.08 μM PTP70-2 was even greater than that of the positive control, 12.5 μM minocycline. In addition to NO, PTP70-2 significantly suppressed the expression of pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Based on these results, the authors believe PTP70-2 is a new anti-neuroinflammatory agent that has great potential for the treatment and prevention of Alzheimer disease.

Lasia spinosa (L.) Thwaites (Araceae) is an ornamental medicinal plant grown in tropical and subtropical countries. The leaf extract of L. spinosa has significant anthelmintic efficacy (Yadav and Temjenmongla 2012). During July 2018 and 2019, L. spinosa plants at the Guangxi Botanical Garden of Medicinal Plants of Guangxi province, China, showed leaves with brown spots or irregular white necrotic lesions that were scattered with some shot holes. Sixty-eight plants from a total of 200 plants showed such symptoms. The overall incidence was about 34%. To isolate the pathogen, 15 infected leaves were excised near the margin of necrotic lesions, surface sterilized with 75% ethanol for 1 min, and treated with 0.5% sodium hypochlorite for 10 min followed with three rinses with sterile water. The leaf sections were then plated on potato dextrose agar (PDA) medium and incubated at 28°C. Further, to obtain pure cultures, hyphal tips from recently germinated spores were excised and transferred onto fresh PDA and incubated at 28°C. Ten fungal isolates with similar morphological characteristics were obtained. Colonies on PDA developed white fluffy aerial mycelia and produced abundant conidia 2 weeks later. Microconidia were abundant, oval, clavate, solitary, and hyaline, with an average size of 7.7 × 3.1 μm (n = 46). Macroconidia were mostly three to five septate, slender, almost straight, and sickle shaped, with an average size of 26.7 × 3.6 μm (n = 30). No chlamydospores were observed. These morphological characteristics were consistent with those of Fusarium fujikuroi (Leslie and Summerell 2006). For further identification, five DNA regions of isolate CYA, corresponding to the internal transcribed spacer (ITS), β-tubulin (TUB2), calmodulin (CL), translation elongation factor 1-alpha (TEF1), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were PCR amplified and sequenced (Jiang et al. 2018; Weir et al. 2012). BLASTN analyses of ITS and CL sequences showed 100% similarity with corresponding sequences of F. fujikuroi accession numbers MK630103.1 and MF984412.1; the TUB2 and TEF1 sequences shared 99.71% identity (MH398245.1 and MH341224.1), and the GAPDH sequence shared 98.65% identity (CP023092.1). Sequences were submitted to GenBank under the accession numbers MN704565 (ITS), MN490089 (TUB2), MN490091 (CL), MN490092 (TEF1), and MN490090 (GAPDH). According to those morphological and molecular characteristics, the pathogen was identified as F. fujikuroi. Further, to verify the pathogenicity, spore suspensions (10⁵ conidia/ml) were prepared from 10-day-old PDA cultures and inoculated onto leaves of nine 2-month-old L. spinosa plants using the wound/drop inoculation method (10 μl per wound). Three more plants were inoculated with sterile water as controls. All plants were grown in a growth chamber with a temperature of 28°C and 90% humidity. After incubating for 7 days, leaf spot symptoms were observed on the inoculated leaves, and no symptoms were observed on the control plants. F. fujikuroi was reisolated from leaf lesions on the inoculated leaves, thus confirming Koch’s postulates. The pathogenic experiment was subsequently repeated under the same conditions. To our knowledge, this is the first report of F. fujikuroi causing leaf spot of L. spinosa. Severe leaf spot disease on L. spinosa has affected its medicinal and ornamental values, which suggests breeders should take this disease into consideration in L. spinosa breeding programs.

This study aimed to demonstrate that prenatal dexamethasone exposure (PDE) can induce kidney dysplasia in utero and adult glomerulosclerosis in male offspring, and to explore the underlying intrauterine programming mechanisms. Pregnant rats were subcutaneously administered dexamethasone 0.2 mg/kg.d from gestational day (GD) 9 to GD20. The male fetus on GD20 and the adult offspring at age of postnatal week 28 were analyzed. The adult offspring kidneys in the PDE group displayed glomerulosclerosis, elevated levels of serum creatinine and urine protein, ultrastructural damage of podocytes, the reduced expression levels of podocyte marker genes, nephrin and podocin. The histone 3 lysine 9 acetylation (H3K9ac) level in the promoter of renal angiotensin II receptor type 2 (AT2R) and its expression were reduced, whereas the angiotensin II receptor type 1a (AT1aR)/AT2R expression ratio was increased. The fetal kidneys in the PDE group displayed an enlarged Bowman's space and a shrunken glomerular tuft, a reduced cortex width and an increase in the nephrogenic zone/cortical zone ratio, reduced the expression level of glial-cell-line derived neurotrophic factor/c-Ret tyrosine kinase receptor (GDNF/c-Ret) signal pathway and podocyte marker genes. Moreover, the H3K9ac and H3K27ac levels of AT2R as well as the gene and protein expression levels of AT2R in fetal kidneys were inhibited by PDE. In vitro, primary metanephric mesenchyme stem cells (MMSCs) were treated with dexamethasone. Overexpression of AT2R reversed the inhibited expression of GDNF/c-Ret and podocin/nephrin induced by dexamethasone, and glucocorticoids receptor antagonist abolished the decreased H3K9ac level and gene expression of AT2R. In conclusion, PDE induced the offspring’s kidney dysplasia as well as adult glomerulosclerosis, which was mediated by a sustained decrease in renal AT2R expression via decreasing the H3 K9ac level.

The objective of this work was to evaluate the effect of poly(lactic acid) (PLA) based biodegradable film packaging combining 0.5% nisin antimicrobial polypeptide on the physicochemical and microbial quality of Boletus edulis wild edible mushrooms stored at 4 ± 1 °C. The experiment was set up by packaging mushrooms with extruded PLA films containing 0, 7.5, and 15 wt.% triethyl citrate plasticizer. The low-density polyethylene (LDPE) film was used as the control. Mushrooms stored in PLA films containing 7.5 and 15 wt.% plasticizer provided better retention of quality characteristics and received higher sensory ratings compared to mushrooms stored in pure PLA film and LDPE film. Samples with these two treatments underwent minimal changes in texture, PPO activity, total bacteria count, and sensory attributes. Results suggest that nisin in combination with plasticized PLA film has the potential to maintain B. edulis wild edible mushroom quality and extend its postharvest life to 18 days. Industrial relevance B. edulis is one of the most commercialized mushrooms worldwide. However, as with all fresh mushrooms, there are severe preservation problems. Extruded PLA films containing triethyl citrate plasticizer plus antimicrobial agent nisin proved to be a suitable technology for mushroom conservation. This material exhibits an environmental-friendliness potential and a high versatility in food packaging.

BF-30 is a single chain polypeptide of an N-segment with an α-helix from cathelicidin gene encoding, and it contains 30 amino acid residues, with a relative molecular mass and isoelectric point of 3637.54 and 11.79, respectively. Cathelicidin-BF-30 was entrapped in four-arm star-shaped poly(ethylene glycol-b-dl-lactic acid-co-glycolic acid) block copolymers (4-arm-PEG-PLGA) by a double-emulsion solvent-evaporation method. Three release phases of cathelicidin-BF-30loaded 4-arm-PEG-PLGA microspheres were observed, including an initial burst-release phase, followed by a lag phase with minimal drug release and finally a secondary zero-order release phase. The delivery system released BF-30 over more than 15 days in vitro. Furthermore, the material for preparing the microspheres has good biocompatibility and biodegradability. Additionally, based on the drug resistance of food pathogenic bacteria, the antibacterial effects of BF-30 on Shigella dysenteriae CMCC 51105 (Sh. dysenteriae CMCC 51105), Salmonella typhi (S. typhi) and Staphylococcus aureus (S. aureus) as well as the stability of the in vitro release of the BF-30-loded microspheres were studied. The α-helix secondary structure and antibacterial activity of released BF-30 were retained and compared with native peptide. These BF-30 loaded microspheres presented

This study aimed to prove that prenatal ethanol exposure (PEE) can induce nephrotic syndrome in male rat offspring and to explore the underlying intrauterine programming mechanisms. Pregnant Wistar rats were intragastrically administered ethanol (4 g/kg d) from gestational day (GD) 9 to GD 20, and the male fetuses were delivered by cesarean section at GD20 and the male adult offspring were euthanized at postnatal week (PW) 24. In vitro, the primary metanephric mesenchyme cells were treated with ethanol at concentrations of 15-60 mM. The results indicated that the kidneys of adult offspring in the PEE group exhibited glomerulosclerosis as well as interstitial fibrosis. The levels of serum creatinine and urine protein were elevated; the serum total cholesterol level was increased and the serum albumin concentration was reduced. In the fetal kidney, developmental retardation was presented in the PEE group via pathological examinations, accompanied by the expressional inhibition of the glial-cell-line-derived neurotrophic factor/c-ret tyrosine kinase receptor (GDNF/c-ret) signaling pathway. Although serum angiotensin II (Ang II) level and the gene expression of renal angiotensin-converting enzyme (ACE) were increased in the PEE group, the expression of renal angiotensin II type 2 receptor (AT2R) was significantly inhibited, accompanied by a reduction in the H3K27ac level on the AT2R gene promoter. In the non-classical renin-angiotensin system (RAS), the expression of renal angiotensin converting enzyme 2 (ACE2) and Mas receptor (MasR) were inhibited in the PEE group. The above changes of the classical and non-classical RAS all sustained from utero to adulthood. In vitro, ethanol elevated the gene expression of ACE and angiotensin II type 1a receptor (AT1aR) whereas it reduced the expression of AT2R, ACE2, and MasR, accompanied by a reduction in the H3K27ac level on AT2R gene promoter. Taken together, these results suggested that PEE can induce fetal kidney developmental retardation and adult nephrotic syndrome, and direct regulation of ethanol to the renal RAS was involved in the mechanism of nephrotic syndrome induced by PEE.

Schizophyllum commune (SC) is widely consumed by Chinese, especially in southern part of China. The aim of the present study was to assess the extract of SC on vascular tone and the mechanisms involved. Experiments were performed on aorta of 18-week-old male Sprague-Dawley rats. Dried SC was extracted with 50% ethanol, 90% ethanol and deionized water, respectively. The effects of SC on the isometric tension of rat aortic rings were measured. Protein expression for the endothelial nitric oxide synthase (eNOS) was also determined in the primarily cultured rat aortic arterial endothelial cells (RAECs). The results showed that the water extract of SC induced a marked relaxation in aortic rings with or without endothelium. After the pretreatments of N(ω)-nitro-l-arginine methyl ester, indomethacin, RP-cAMP, and methylene blue, the SC-induced relaxation was significantly decreased. In addition, the contraction due to Ca(2+) influx and intracellular Ca(2+) release was also inhibited by SC. Furthermore, expression of the eNOS protein was significantly elevated in RAECs after treatment of SC. In conclusion, the water extract of SC induces an endothelium-dependent and -independent relaxation in rat aorta. The relaxing effect of SC involves the modulation of NO-cGMP-dependent pathways, PGI2-cAMP-depedent pathways, Ca(2+) influx though calcium channels and intracellular Ca(2+) release.

Rubella viruses of genotypes 1E and 2B are currently the most frequently detected wild-type viruses in the world. Genotype 1E viruses from China have been genetically distinct from genotype 1E viruses found elsewhere, while genotype 2B viruses found in China are not distinguishable from genotype 2B viruses from other areas. Genetic clusters of viruses of both genotypes were defined previously using sequences of the 739-nt genotyping window. Here we report phylogenic analysis using whole genomic sequences from seven genotype 1E and three genotype 2B viruses which were isolated in China between 2000 and 2013 and confirm the subgrouping of current circulating genotypes 1E and 2B viruses. In addition, the whole genomic characterization of Chinese rubella viruses was clarified. The results indicated that the Chinese rubella viruses were highly conserved at the genomic level, and no predicted amino acid variations were found at positions where functional domains of the proteins were identified. Therefore, it gives us the idea that the rubella control and elimination goal should be achieved if vaccine immunization coverage continues maintaining at the high level.

The incidence of rubella cases in China from 1991 to 2007 was reviewed, and the nucleotide sequences from 123 rubella viruses collected during 1999 to 2007 and 4 viral sequences previously reported from 1979 to 1984 were phylogenetically analyzed. Rubella vaccination was not included in national immunization programs in China before 2007. Changes in endemic viruses were compared with incidences of rubella epidemics. The results showed that rubella epidemics occur approximately every 6 to 8 years (1993/1994, 2001, and 2007), and a shift of disease burden to susceptible young adults was observed. The Chinese rubella virus sequences were categorized into 5 of the 13 rubella virus genotypes, 1a, 1E, 1F, 2A, and 2B; cocirculations of these different genotypes were found in China. In Anhui province, a shift in the predominant genotype from 1F and 2B to 1E coincided with the 2001 rubella epidemic. This shift may have occurred throughout China during 2001 to 2007. This study investigated the genotype distribution of rubella viruses in China over a 28-year period to establish an important genetic baseline in China during its prevaccination era.

We explored the effects on brain oedema and neurological functional recovery after transplantation of hAECs (human amniotic epithelial cells) into the lateral ventricle of rats with ICH (intracerebral haemorrhage). hAECs were isolated from human term placenta and seeded for primary culture. We delivered hAECs labelled with Hoechst33258 and transfected with EGFP (enhanced green fluorescent protein) gene using lentiviral vectors into ICH rat models. The behaviour of the animals and brain oedema were evaluated after 28 days, and brain sections were made for morphological and immunohistochemical analyses with fluorescence microscopy. Our results were as follows. Transplanted hAECs were observed along the lateral wall and survived for at least 4 weeks. Some of the cells were stained with human specific antibody to vimentin and nestin. Around the injury site, activated microglia stained with OX42 were reduced. The water content of ICH rats decreased in the treatment group. The behaviour test scores were improved in the treatment group compared with those in the control groups. In conclusion, hAECs cannot only survive in the lateral ventricle of ICH rats after transplantation, but also express vimentin and nestin. hAEC transplantation reduced brain oedema and improved the motor deficits of ICH rats.

The combination of ATO and bortezomib (ATO+bortezomib) has been recently shown to enhance antimyeloma activity; nevertheless, the mechanisms remained unclear in these studies. However, both bortezomib and ATO have been shown to activate the p38 MAPK pathway, which may counteract the enhancement induced by this combination. We studied the cytotoxicity of bortezomib, ATO, and ATO+bortezomib with or without inhibiting p38 MAPK, along with associated molecular changes in myeloma cells. The treatment of myeloma cells with ATO+bortezomib induced higher cytotoxicity than either agent alone. This increased cytotoxicity was further synergistically enhanced by inhibiting p38 MAPK. This effect was preserved in the presence of marrow stromal cells designed to simulate the tumor micro-environment and in the CD138+ neoplastic plasma cells directly isolated from myeloma patients. The enhanced cytotoxicity of ATO+bortezomib was associated with augmented STAT3 inhibition and JNK activation, up-regulation of Bim, p21, p27, p53 as well as down-regulation of Bcl-2. Furthermore, the synergistically potentiated apoptosis by p38 MAPK inhibition was associated with the attenuation of ATO+bortezomib-mediated activation of Hsp27 as well as the enhancement of ATO+bortezomib-mediated JNK activation, p53 up-regulation, and Bcl-2 down-regulation. The results suggest the opportunity for using p38 MAPK inhibition to enhance the efficacy of ATO+bortezomib in myeloma.

Alzheimer's disease is a progressive neurodegenerative disorder, characterized by irreversible impairment of memory and cognitive function. The exact causes of Alzheimer's disease still remain unclear and current single target drugs could only offer limited therapeutic effect to the patients. We have previously reported that T-006, a promising anti-Alzheimer's compound derived from Chinese medicinal component tetramethylpyrazine, might protect neurons through inhibiting the overproduction of intracellular reactive oxygen species (ROS) and reactive nitrogen species (RNS). In this study, we further investigated the neuroprotective effects, as well as the molecular pathways involved, of T-006 against glutamate-induced excitotoxicity in rat cerebellar granule neurons (CGNs). T-006 was also found to promote neuronal differentiation in both PC12 cells and primary cultured rat cortical neurons. The results showed that the pretreatment of T-006 (0.01-1 μM) might prevent glutamate-induced neuronal loss in a concentration-dependent manner. T-006 is found to inhibit the over-activation of NMDAR and ensued calcium overload caused by glutamate. The following activation of phosphorylated extracellular signal-regulated kinase (ERK) were also abolished. Moreover, T-006 concurrently prevented the suppression of phosphorylated protein kinase B (Akt) and glycogen synthase kinase 3β (GSK3β). T-006 was also found to promote neurite outgrowth in PC12 cells and primary cortical neurons. In our study, T-006 (0.1-3 μM) dose-dependently stimulated neurite outgrowth in PC12 cells and the efficacy was comparable to nerve growth factor (NGF). Moreover, co-treatment of T-006 and NGF revealed that T-006 could robustly potentiate the NGF-induced neuritogenesis. Further signal transduction studies indicated that T-006 rapidly up-regulated phosphorylation of ERK but did not activate tyrosine kinase receptor A (Trk A). These findings offer deeper understanding of the anti-neurodegenerative activity of T-006 and provide insight into its possible therapeutic potential for AD treatment in light of the multipotent nature of T-006.

Dear Editor, Anthrax is an acute zoonotic infectious disease that is caused by Bacillus anthracis. Humans and other animals are susceptible to anthrax. Cattle, horses, sheep, camels and other herbivores are the main sources of infection in human anthrax cases. Anthrax infection in humans can occur through contact with sick animals or their products.1 Patients with cutaneous anthrax may present with skin necrosis, ulceration, extensive edema in the surrounding tissues, toxemia and other symptoms. Other forms of the disease include pulmonary anthrax caused by inhalation and intestinal anthrax caused by ingestion.2, 3 Because anthrax is very contagious, has severe manifestations, and can have an abrupt onset, every country classifies anthrax as a highly infectious disease.1, 4 Moreover, since the ‘anthrax powder' terrorist event that occurred in the United States in 2001, there has been a global concern regarding anthrax bioterrorism.5 This report describes an outbreak of anthrax in humans that occurred in Yunnan province, China, in 2015 following the slaughter of cows. On October 30, 2015, the outpatient department of infectious diseases in the third hospital of Kunming (hereafter referred to as the hospital), received two cases (cases 1 and 2 in Table 1) in the morning and two more cases (cases 3 and 4 in Table 1) later that day. Blisters of different sizes were observed on both the hands and arms of each patient. All four patients were from the same village in Dongchuan, Kunming, Yunnan Province, China (25°57′ north latitude, 102°47′ east longitude). Dongchuan is located 150 km from Kunming. Table 1 Clinical manifestations and laboratory test results for four cutaneous anthrax cases By questioning the patients, the doctor learned that the four patients worked at a beef restaurant located in Dongchuan. The owner of the restaurant, who was one of the four patients, had purchased a head of sick cattle that was unable to walk from a local farmer at a cheap price, and it was immediately slaughtered at the restaurant. The four patients had aided in slaughtering the cattle on October 19, 2015. The slaughter commenced in the morning and was completed at noon on the same day. The beef and viscera were cooked for use in meals at the restaurant (the local villagers habitually eat beef). From 21 October, which was the third day following the slaughter, until 28 October, blisters appeared on the hands or arms of the four restaurant employees who had participated in the slaughter. Because no significant fever or other discomfort was noted, the affected individuals' blisters were dressed at a local clinic, and no other treatment was administered. As time passed, the number of blisters on the patients' arms increased, and all four patients experienced increasing pain. Therefore, a local doctor recommended that the patients seek treatment at the hospital in Kunming. A body surface examination of each patient revealed diffuse blisters of various sizes (with diameters ranging from 0.5 cm to 5 cm) on the left or right hands, back or arms. Blister-like skin lesions were noted on the entire right arm in the fourth case. A yellowish exudate was observed in the lesions, the tissues surrounding the blisters were swollen, and some of the blisters resembled creamy white pustules, with swelling of the corresponding arms. The findings from our examinations and the degree of skin damage to the arms in the four cases are shown in Table 1. Because the four cases had a common history of exposure to cattle (perhaps infected cattle) and exhibited signs of anthrax (for example, vesicular skin lesions), the attending physician determined that the patients could have been infected with B. anthracis during the slaughter of the cow at the patients' place of employment. To avoid further spread of the disease, the hospital immediately transferred the four patients from the outpatient ward to the isolation ward, which is located in a different building. The isolation ward was designed to accommodate patients with major infectious diseases (for example, severe acute respiratory syndrome and human avian influenza), so the four patients in this ward were kept isolated from each other. The doctors and nurses maintained proper physical protection against infection during the diagnosis and treatment of the four cases. The medical staff bandaged the damaged skin of the patients to avoid further exposure to infection due to exudation. Routine blood tests revealed that neutrophils accounted for more than 85% of the white blood cell count in three of the four patients (Table 1). The hospital immediately reported the disease to the local center for disease control and prevention (CDC). The CDC collected samples of the exudate from the damaged skin of the patients in the hospital and carried out bacterial smears and cultures. On 31 October, the hospital received the CDC's report, which indicated that the bacterial smears and cultures for two of the four patients were positive for B. anthracis. On the basis of the patients' epidemiological information, clinical manifestations and positive anthrax test results, the doctors determined that the four patients had cutaneous anthrax due to infection with B. anthracis that was transmitted during the slaughter of ailing cattle. Although the patients were hospitalized, they were stable, and no pulmonary infection, nausea, vomiting or other complications occurred. After treatment, the edema in the patients' arms where skin had been damaged gradually subsided. In addition, diminished exudation was noted from the original blisters, and black scabs formed. On 17 November, the local CDC performed anthrax smears and bacterial cultures using new liquid samples collected from the four patients' damaged skin, and all of the results were negative. The four patients were discharged from the hospital on 20 November, 2015. Anthrax is a natural focal disease with global distribution. Anthrax is most commonly found in pastoral areas of South America, Asia and Africa.6 Anthrax is a notifiable infectious disease in China, and each case must be promptly reported to health authorities.7 A total of 1911 cases of cutaneous anthrax were reported in 225 counties within 18 provinces from 2007 to 2011. Of these cases, 13 were fatal.8 Most of the cases were middle-aged individuals. Herders and farmers accounted for 86.08% of the total cases. In recent years, the number of anthrax cases has decreased in China.9 Anthrax cases in China are concentrated mainly in pastoral areas with poor economic and health conditions. Local herders are often unaware of how anthrax is transmitted and of the dangers of the disease; therefore, they are reluctant to discard dead animals and may skin them on-site for meat, resulting in infection. Cattle farming in Yunnan is common, and a large number of cattle is bred there. In addition, Yunnan is a natural focus of anthrax and several anthrax outbreaks related to the slaughter of cattle have been reported in recent years.9 The four reported cases in this study acquired anthrax during the slaughter of cattle. Because of timely treatment, the four patients fully recovered in a short time without complications. All of the patients returned home after treatment. In addition, the attending doctors quickly isolated the affected patients from the other patients at the hospital, thereby preventing the occurrence of secondary cases and further spread of the epidemic. As mentioned above, anthrax infections have occurred in some places in China. Until now, anthrax has been reported in mountainous areas, where cattle, sheep and other livestock are kept.10 Improving the management of sick animals and enabling herders to handle sick animals correctly by providing them with proper educational materials will help significantly reduce the number of human cases of anthrax due to contact with infected animals.

In China, rubella vaccination was introduced into the national immunization program in 2008, and a rubella epidemic occurred in the same year. In order to know whether changes in the genotypic distribution of rubella viruses have occurred in the postvaccination era, we investigate in detail the epidemiological profile of rubella in China and estimate the evolutionary rate, molecular clock phylogeny, and demographic history of the predominant rubella virus genotypes circulating in China using Bayesian Markov chain Monte Carlo phylodynamic analyses. 1E was found to be the predominant rubella virus genotype since its initial isolation in China in 2001, and no genotypic shift has occurred since then. The results suggest that the global 1E genotype may have diverged in 1995 and that it has evolved at a mutation rate of 1.65 × 10 −3 per site per year. The Chinese 1E rubella virus isolates were grouped into either cluster 1 or cluster 2, which likely originated in 1997 and 2006, respectively. Cluster 1 viruses were found in all provinces examined in this study and had a mutation rate of 1.90 × 10 −3 per site per year. The effective number of infections remained constant until 2007, and along with the introduction of rubella vaccine into the national immunization program, although the circulation of cluster 1 viruses has not been interrupted, some viral lineages have disappeared, and the epidemic started a decline that led to a decrease in the effective population size. Cluster 2 viruses were found only in Hainan Province, likely because of importation.