Mechanical pain is one of the most common causes of clinical pain, but there remains a lack of effective treatment for debilitating mechanical and chronic forms of neuropathic pain. Recently, neurotoxin GsMTx4, a selective mechanosensitive (MS) channel inhibitor, has been found to be effective, while the underlying mechanism remains elusive. Here, with multiple rodent pain models, we demonstrated that a GsMTx4-based 17-residue peptide, which we call P10581, was able to reduce mechanical hyperalgesia and neuropathic pain. The analgesic effects of P10581 can be as strong as morphine but is not toxic in animal models. The anti-hyperalgesic effect of the peptide was resistant to naloxone (an μ-opioid receptor antagonist) and showed no side effects of morphine, including tolerance, motor impairment, and conditioned place preference. Pharmacological inhibition of TRPV4 by P10581 in a heterogeneous expression system, combined with the use of Trpv4 knockout mice indicates that TRPV4 channels may act as the potential target for the analgesic effect of P10581. Our study identified a potential drug for curing mechanical pain and exposed its mechanism.

OBJECTIVE To analyze the metabolites of Zhideke granules and speculate its metabolic pathway in rats in vivo. METHODS Male SD rats were randomly divided into blank group and administration group （Zhideke granules， 9.45 g/kg）； they were given ultrapure water or relevant medicine， twice a day， every 6-8 h， for 3 consecutive days. Serum， urine and feces samples of rats were collected， and their metabolites were identified by UPLC-Q-Exactive-MS technique after intragastric administration of Zhideke granules； their metabolic pathways were speculated. RESULTS After intragastric administration of Zhideke granules， 16 prototype components （i.g. irisflorentin， baicalin， chlorogenic acid） and 11 metabolites （i.g. hydration products of kaempferol or luteolin， methylation products of chlorogenic acid， and hydroxylation products of baicalin） were identified in serum， urine and feces of rats. Among them， 8 prototype components and 4 metabolites were identified in serum samples； 10 prototype components and 7 metabolites were identified in urine samples； 8 prototype components and 5 metabolites were identified in the fecal samples. CONCLUSIONS The metabolites of Zhideke granules in rats mainly include baicalin， irisflorentin，chlorogenic acid， and the main metabolic pathways included methylation， hydroxylation， glucuronidation.

OBJECTIVE To provide reference for basic analysis of the pharmacodynamic substance in Stahlianthus involucratus. METHODS Overall 24 SD male rats were randomly divided into blank group （purified water）， and administration group （ethanol extract of S. involucratus， 15.75 g/kg， calculated by crude drug）， with 12 rats in each group. They were given drug liquid/purified water intragastrically， twice a day， every 6-8 h， for consecutive 3 days. After medication， the blood， urine and fecal samples were collected from two groups of rats. UPLC-Q-Exactive-MS technology was used to identify the chemical constituents in the ethanol extract of S. involucratus， and metabolites in the blood， urine and fecal of rats after intragastrical administration of the ethanol extract of S. involucratus. Multivariate statistical analysis was employed to screen various serum metabolites. Metabolic pathways were analyzed by MetaboAnalyst 5.0 platform. RESULTS A total of 38 chemical constituents were identified from the ethanol extract of S. involucratus， including fourteen prototype components and three metabolites identified from 5 urine samples， nine prototype components identified from fecal samples， and ten prototype components and one metabolite identified from serum samples. A total of 71 differential metabolites were screened from two groups of rat serum samples， of which 44 differential metabolites， such as ferulic acid， glycyrrhizin， were up-regulated and 27 differential metabolites， such as arachidonic acid， phenylacetylglutamine， were down-regulated. The 71 differential metabolites were mainly enriched in 11 metabolic pathways， including phenylalanine metabolism， linoleic acid metabolism， arachidonic acid metabolism， and tryptophan metabolism. CONCLUSIONS Ferulic acid， liquiritigenin， isofraxidin and formononetin may be the material basis that directly exert pharmacological effects of S. involucratus. S. involucratus may exert anti-inflammatory effects by affecting metabolic pathways， including arachidonic acid metabolism and tryptophan metabolism.

Objective:To observe the effect of Nodal on the biological behavior of retinal vascular endothelial cells (RF/6A cells) in monkeys with high glucose.Methods:RF/6A cells were divided into normal group, mannitol group, high glucose group, high glucose combined with non-specific small interfering RNA treatment group (HG+NC group), high glucose combined with small interfering Nodal treatment group (HG+siNodal group). The transfection efficiency of siNodal was observed by real-time fluorescence quantitative PCR and western blot protein immunoblotting. The effect of Nodal on the proliferation of RF/6A cells was detected by thiazole blue colorimetry. The effect of Nodal on migration ability of RF/6A cells was detected by cell scratch assay. The effect of Nodal on the formation of RF/6A cell lumen was measured by Matrigel three-dimensional in vitro. The expression of extracellular signal phosphorylated regulated kinase 1/2 (pERK1/2) in RF/6A cells was detected by western blot protein immunoblotting. One-way analysis of variance was used to compare groups.Results:Compared with HG+NC group, Nodal protein ( F=33.469) and mRNA relative expression levels ( F=38.191) in HG+siNodal group were significantly decreased, cell proliferation was significantly decreased ( F=28.548), and cell migration ability was significantly decreased ( F=24.182). The number of cell lumen formation was significantly decreased ( F=52.643), and the differences were statistically significant ( P<0.05). Compared with HG+NC group, the relative expression of pERK1/2 protein in HG+siNodal group was significantly decreased, and the difference was statistically significant ( F=44.462, P<0.01). Conclusions:Silencing Nodal expression can inhibit proliferation, migration and tube formation of RF/6A cells induced by high glucose. It may act by inhibiting pERK1/2 expression.

Objective:To observe the effect of bone forming protein 4 (BMP4) on the proliferation and migration of human retinal pigment epithelium (RPE) cells under oxidative stress, and to preliminarily explore its effect on epithelial-mesenchymal transition (EMT) of RPE cells.Methods:Human RPE cells cultured in vitro were divided into normal group, pure 4-hydroxynonenal (HNE) group (4-HNE group), 4-HNE+NC group and 4-HNE+ small interfering BMP (siBMP4) group. The effect of 4-HNE on the proliferation of RPE cells was detected by thiazole blue colorimetry. The effects of 4-HNE and BMP4 on cell migration were determined by cell scratch test. The expression of BMP4 was detected by immunofluorescence staining, Western blot and real-time quantitative polymerase chain reaction. The transfection efficiency of siBMP4 was observed by fluorescence microscopy. Mitochondrial reactive oxygen species (MitoSOX) were detected by flow cytometry. The expression of EMT markers E-cadherin and Fibronection were detected by immunofluorescence assay. t-test was used for comparison between the two groups, and one-way analysis of variance was used for comparison between the three groups. Results:Compared with normal group, cell proliferation and migration ability of 4-HNE group were significantly enhanced, with statistical significance ( t=21.619, 24.469; P<0.05). The expression of BMP4 in cells was significantly increased, and the difference was statistically significant ( t=19.441, P<0.05). The relative expression levels of BMP4 mRNA and protein were also significantly increased, with statistical significance ( t=26.163, 37.163; P<0.05). After transfection with siBMP4 for 24 h, the transfection efficiency of BMP4 in RPE cells was>90%. Compared with 4-HNE group and 4-HNE+NC group, the relative expression levels of BMP4 protein ( F=27.241), mRNA ( F=36.943), cell mobility ( F=46.723) and MitoSOX expression levels ( F=39.721) in normal group and 4-HNE+siBMP4 group were significantly decreased. The differences were statistically significant ( P<0.05). The epithelial marker E-cadherin increased significantly, while the mesenchymal marker Fibronection decreased significantly, with statistical significance ( F= 51.722, 45.153; P<0.05). Conclusions:BMP4 inhibits RPE proliferation and migration under oxidative stress. BMP4 is involved in inducing EMT in RPE cells.

Liver fibrosis is a wound healing response that occurs in the setting of chronic liver injury and is caused by imbalance in the synthesis and degradation of extracellular matrix （ECM）. If left untreated， it can progress to liver cirrhosis and hepatocellular carcinoma. The activation of hepatic stellate cell （HSC） is now well established as a central driver of liver fibrosis. The activated HSC will transform into myofibroblasts that produce ECM protein. Transforming growth factor-β₁ （TGF-β₁） can induce the activation of hepatic stellate cell （HSC）， and TGF-β₁/Smads signaling pathway is one of the important pathways to promote liver fibrosis. Non-coding RNA （ncRNA） does not encode proteins during the transcription but plays an important regulatory role in the post-transcriptional process of genes. Accumulating evidence shows that the occurrence of liver fibrosis is closely related to the abnormal expression of ncRNA which participates in the activation of HSC by regulating TGF-β₁ signal transduction and then affects the process of liver fibrosis. MiRNA-mediated TGF-β₁/Smads signaling pathway can not only promote liver fibrosis but also play a role in anti-fibrosis. Long non-coding RNA （lncRNA） not only promotes the development of liver fibrosis by binding to target genes but also enhances TGF-β₁ signal transduction by acting as competitive endogenous RNA. circular RNA （circRNA） acts as a ''sponge'' to regulate TGF-β₁/Smads pathway， thereby inhibiting HSC activation and exerting the anti-liver fibrosis effect. Chinese medicinal plays an essential part in the prevention and treatment of liver fibrosis， and the active components can inhibit TGF-β₁/Smads pathway by regulating the expression of miRNA， thus alleviating liver fibrosis. This article reviews the role and mechanism of miRNA-， lncRNA- and circRNA-mediated TGF-β₁/Smads signaling pathway in liver fibrosis and summarizes the anti-liver fibrosis effect of active components of Chinese medicinals by regulating miRNA-mediated TGF-β₁/Smads signaling pathway， which can serve as a reference for clinical treatment of liver fibrosis and the development of new drugs.

Objective:To explore the potential genetic causes of unexplained neonatal encephalopathy.Methods:This retrospective study enrolled 113 infants diagnosed with unexplained neonatal encephalopathy and underwent genetic testing in the Children's Hospital of Hunan Province from January 2019 to May 2021. Perinatal data, clinical manifestations, electroencephalograph, brain MRI findings, genetic information, and prognosis of those patients were analyzed. T-test or Chi-square test were used for data analysis. Results:Of the 113 infants enrolled, 74 (65.5%) were males. The gestational age at birth was (38.6±1.5) weeks, and the birth weight was (2 957±561) g. The most common clinical manifestation was the disturbance of consciousness (83/113, 73.5%), followed by seizures (39/113, 34.5%). There were 38.2% (34/89) of the patients with abnormal brain MRI, and 80.4% (74/92) presented abnormal electroencephalography. Among the 113 infants, 60 (53.1%) had genetic abnormalities, including 48 with single nucleotide variations, eight with copy number variations, and four with chromosome abnormalities. Single nucleotide variations in the 48 patients were classified into syndromic ( n=18, 37.5%), metabolic ( n=16, 33.3%), epileptic ( n=11, 22.9%) and mitochondrial-related genes ( n=3, 6.3%), of which 14 were not included in any database. Among the 103 cases which were successfully followed up until December 31, 2021, 75 (72.8%) had a poor prognosis, including 52 (50.5%) death cases and 23 (22.3%) cases of development retardation. Birth weight and the incidence of seizures in the poor prognosis group were both lower than those in the non-poor prognosis group [(2 876±536) vs (3 254±554) g, t=3.15; 29.3% (22/75) vs 53.6% (15/28), χ2=5.20; both P<0.05], while the incidence of disturbance of consciousness was higher [80.0% (60/75) vs 53.6% (15/28), χ2=7.19, P<0.05]. The proportion of infants with genetic abnormalities in the poor prognosis group was higher than that in the non-poor prognosis group, but the difference was not statistically significant [53.3% (40/75) vs 46.4% (13/28), χ2=0.39, P=0.533]. Conclusions:Genetic abnormality is one of the leading causes of unexplained neonatal encephalopathy. Nucleotide variation is the most common genetic type. Syndromic, metabolic, and epileptic variants are frequently detected in these patients.

Objective:To isolate and culture WU polyomavirus (WUPyV), and to analyze the genome-wide evolutionary patterns, homology and population dynamics.Methods:Real-time quantitative PCR was used to detect the nasopharyngeal aspirate samples of hospitalized children with respiratory tract infection in Beijing Friendship Hospital during 2020 to 2022. Primary human airway epithelial cells cultured at the air-liquid interface were used to isolate and culture WUPyV. Whole genome sequence of the isolated strain was obtained by Sanger sequencing. For phylogenetic and evolutionary dynamics analysis, the whole genome was compared with the published whole genome sequences in GenBank database.Results:The detection rate of WUPyV was 4.7% (31/659) during 2020 to 2022, and a clinical strain BJ0593 of WUPyV type Ⅲc was successfully isolated. The homology of the whole genome and gene fragments of WUPyV was high. The average evolutionary rate of VP2 gene was about 1.256×10 -4 substitution/site every year, and the population dynamics of WUPyV tended to be flat in the last decade. Conclusions:This study successfully isolated a clinical WUPyV type Ⅲ strain for the first time, which provided the basis for further investigation on the molecular evolution and pathogenicity of WUPyV.

OBJECTIVE To establish a drug batch number traceability management mode in intelligent anesthesia pharmacy of operating room， improve the management level of batch number traceability in anesthesia pharmacy of operating room and ensure the safety of surgical medication. METHODS The problems encountered in the application of batch number traceability management mode in intelligent anesthesia pharmacy of operating room were analyzed to implement the optimization measure and summarize the optimization effect. RESULTS In view of the incompatibility of multiple batch numbers on the dosing track， the insufficient number of drugs in the automated dispensing cabinet， the long replenishment time， the low stability of the automated dispensing cabinet， and the tedious management of drugs with a near-expiration period， the following measures were implemented， including changing the minimum unit of dosing， adjusting the drug base number and surgery schedule， optimizing the supplementary drug delivery order， repairing dosing tracks of the automated dispensing cabinet， adding the batch number correction function， improving the screening function of drugs with a near-expiration period， and configuring the automated dispensing cabinet in the recovery room. It realized the coexistence of multiple batch numbers on the dosing track， the sufficient number of drugs in the automated dispensing cabinet， the shortening of the replenishment time， the improvement in the stability of the automated dispensing cabinet， and the efficient management of drugs with a near-expiration period. CONCLUSIONS The batch number traceability management mode is established in intelligent anesthesia pharmacy of operating room in our hospital， which improves the system function module， optimizes the work process， increases the stability of the automated dispensing cabinet， improves the work efficiency， and realizes the refined management of surgical medication.

The application of intraocular drug delivery is usually limited due to special anatomical and physiological barriers, and the elimination mechanisms in the eye. Organic nano-drug delivery carriers exhibit excellent adhesion, permeability, targeted modification and controlled release abilities to overcome the obstacles and improve the efficiency of drug delivery and bioavailability. Solid lipid nanoparticles can entrap the active components in the lipid structure to improve the stability of drugs and reduce the production cost. Liposomes can transport hydrophobic or hydrophilic molecules, including small molecules, proteins and nucleic acids. Compared with linear macromolecules, dendrimers have a regular structure and well-defined molecular mass and size, which can precisely control the molecular shape and functional groups. Degradable polymer materials endow nano-delivery systems a variety of size, potential, morphology and other characteristics, which enable controlled release of drugs and are easy to modify with a variety of ligands and functional molecules. Organic biomimetic nanocarriers are highly optimized through evolution of natural particles, showing better biocompatibility and lower toxicity. In this article, we summarize the advantages of organic nanocarriers in overcoming multiple barriers and improving the bioavailability of drugs, and highlight the latest research progresses on the application of organic nanocarriers for treatment of ocular diseases.
Drug Carriers ; Delayed-Action Preparations ; Drug Delivery Systems ; Nanoparticles/chemistry*