Natural products (NPs) are an important source of new drugs for the treatment of stroke. Identifying cellular targets for bioactive molecules is a major challenge and critical issue in the development of new drugs for stroke. Small-molecule probes play a unique role in target discovery. However, drawbacks to these probes include non-specificity, unstable activity, and difficulty in synthesis. Small-molecule probes based on NPs at least partially compensate for these shortcomings. NPs feature rich chemical and structural diversity, biocompatibility, and unique biological activities. These features could be exploited to provide new ideas and tools for target discovery. Small-molecule probes based on NPs provide a precise and direct search for interacting protein targets of NPs-active small molecules. This review explores the properties of small-molecule probes based on NPs and their applications in mechanistic studies of stroke and other diseases. We hope that this review will bring new perspectives to the mechanistic study of NPs-active small molecules and accelerate the translation of these ingredients into drug candidates for the treatment of stroke.

In this study, 34 deep eutectic solvents (DESs) were successfully prepared for the extraction of proanthocyanidin from Rhodiolae Crenulatae Radix et Rhizomes. The extraction process was optimized using single factor exploration and Box-Behnken design-response surface analysis. The extraction rate was significantly improved when the molar ratio of choline chloride to 1,3-propanediol was 1:3.5 and the water content was 30% (V/V) in DESs. AB-8 macroporous resin and ethyl acetate were used for separation and refining, and the oligomer-rich proanthocyanidin components were eventually obtained. The ultraviolet (UV) and infrared (IR) spectra showed that the proanthocyanidins were mainly composed of catechin and epicatechin. To further clarify the chemical composition of proanthocyanidin, an ion scan list containing 156 proanthocyanidins precursors was obtained by constructing a proanthocyanidins structural library and mass defect filtering (MDF) algorithm, combined with the full mass spectrometry (MS)/dd-MS² scan mode that turns on the "if idle pick others" function. By using ultra-high performance liquid chromatography and high-resolution MS (UHPLC/HRMS), the analysis used both targeted and non-targeted methods to detect proanthocyanidins. Finally, 50 oligomeric proanthocyanidin (OPC) compounds were identified, including 7 monomers, 22 dimers, 20 trimers, and 1 tetramer, most of which were procyanidins of proanthocyanidins (84%), and a small amount of prodelphinidin (14%) and other types of proanthocyanidins (2%), which enabled the systematic characterization of proanthocyanidin components from Rhodiolae Crenulatae Radix et Rhizomes. Meanwhile, the comparison with the grape seeds OPCs standard (United States Pharmacopeia) revealed that the proanthocyanidins in Rhodiolae Crenulatae Radix et Rhizomes were more abundant, suggesting that the proanthocyanidins in Rhodiolae Crenulatae Radix et Rhizomes has promising applications.

For wild natural medicine, unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials, which affects the efficacy and safety of clinical medication. DNA barcoding as an effective species identification tool is limited by its low sample throughput nature. In this study, combining DNA mini-barcode, DNA metabarcoding and species delimitation method, a novel biological sources consistency evaluation strategy was proposed, and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as "Guang Dilong" and 25 batches of proprietary Chinese medicines. Besides Amynthas aspergillum as the authentic source, 8 other Molecular Operational Taxonomic Units (MOTUs) were elucidated. Significantly, even the subgroups within A. aspergillum revealed here differ significantly on chemical compositions and biological activity. Fortunately, this biodiversity could be controlled when the collection was limited to designated areas, as proved by 2796 "decoction pieces" samples. This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control, and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.

ObjectiveTo observe the effect of bilateral sequential repetitive transcranial magnetic stimulation on the motor function of upper limbs in stroke patients. MethodsFrom December, 2020 to December, 2022, 62 stroke inpatients in the Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine were included. They were randomly divided into control group (n = 31) and observation group (n = 31). Both groups accepted conventional medicine and rehabilitation, as well as electroacupuncture antagonistic muscle therapy. Before electroacupuncture, the observation group acceped low-frequency repetitive transcranial magnetic stimulation at primary motor cortex (M1) on the healthy side, followed by intermittent theta burst stimulation at M1 on the affected side, for four weeks. They were assessed with Fugl-Meyer Assessment-Upper Extremities (FMA-UE), modified Barthel Index (MBI) and modified Ashworth Scale (MAS), and motor evoked potential (MEP) latency was compared before and after treatment. ResultsOne patient dropped down in the observation group, and no adverse event happened. After treatment, the scores of FMA-UE and MBI significantly increased (|t| > 9.953, P < 0.001), and the score of MAS and the latency of MEP significantly decreased (|t| > 5.043, P < 0.001) in both groups; while all of them were better in the observation group than in the control group (|t| > 2.237, P < 0.05). ConclusionBilateral sequential repetitive transcranial magnetic stimulation can effectively promote the recovery of upper limb motor function in stroke patients.

ObjectiveTo explore the current status, hot spots and development of researches on post-stroke dysphagia in recent five years with visualization analysis. MethodsResearches on post-stroke dysphagia were retrieved from Web of Science Core Collection, from January 1st, 2017 to December 31st, 2021, and reviewed with CiteSpace 5.7.R5 software. ResultsA total of 857 articles were included. The annual number of articles increased with time, published from 61 countries/regions, by 211 institutions and 247 authors. The United States, Japan and China were the most prolific countries, and the University of Manchester was the leading institution. The top three authors were Dziewas R, Wakabayashi H and Hamdy S. The keywords bursting in recent three years included prediction, severity, percutaneous endoscopic gastrostomy, accuracy and early management; co-word clustering showed that researches might be mainly about neuromuscular electrical stimulation, swallowing screening, stroke-associated pneumonia, evidence-based practice, etc. ConclusionThe researches in the field of post-stroke dysphagia have been increasing.

Objective:To analyze the causes and epidemiological characteristics of an outbreak of cutaneous anthrax in Huanan County of Jiamusi City, Heilongjiang Province, so as to provide scientific basis for prevention and control of anthrax.Methods:According to the "Anthrax Epidemiological Case Investigation Form", the field epidemiological investigation was used to investigate the anthrax cases reported in the disease surveillance information report management system (including epidemiological contact history and close contact information); the wound secretion and blood sample anthrax case of and tissue samples of diseased cattle were collected for laboratory testing ( Bacillus anthracis smear microscopy, bacterial isolation and culture, enzyme-linked immunosorbent assay). Results:A total of 8 cases of cutaneous anthrax were reported in this epidemic, including 6 clinically diagnosed cases and 2 confirmed cases (1 death), all of whom were farmers. The diseased cattle tissue samples were examined by smear microscopy and bacterial isolation and culture testing, and were identified as bovine anthrax, the source of the epidemic was determined to be dead cattle; the mode of transmission was mainly direct contact infection. A total of 228 people were in contact with the epidemic, 19 directly involved in peeling and segmentation, and 11 in transportation and loading and unloading, all of which were included as the key observation subjects; 198 people who had the behaviors of buying, processing, eating and storage were all included as the general observation subjects.Conclusions:Slaughtering and contacting infected cattle by key occupational groups are the main reasons for the epidemic. Health education should be strengthened for key populations in rural aquaculture industry to prevent and control the occurrence of anthrax.

Objective:To compare the therapeutic effect of fluid resuscitation strategy guided by pulse-indicated continuous cardiac output (PiCCO) monitoring and early goal-directed therapy (EGDT) on renal function of acute kidney injury (AKI) patients caused by septic shock.Methods:Septic shock patients with AKI admitted to the intensive care unit (ICU) of Tianjin Fifth Central Hospital and Teda International Cardiovascular Hospital from March 2017 to February 2020 were enrolled. All patients were given fluid resuscitation. Patients were divided into PiCCO-guided fluid resuscitation group [PiCCO group, intrathoracic blood volume index (ITBVI) was maintained between 850-1 000 mL/m 2] and EGDT-guided fluid resuscitation group [EGDT group, central venous pressure (CVP) was maintained between 8-12 mmHg (1 mmHg = 0.133 kPa) or CVP ≤ 15 mmHg when patients received mechanical ventilation (MV)] according to both the patient's condition and the informed consent of the patient's family. The changes of heart rate (HR), mean arterial pressure (MAP), CVP, blood lactic acid (Lac), fluid balance, urine volume and serum creatinine (SCr) at 6, 24, and 48 hours after fluid resuscitation in the two groups were observed, and the renal replacement therapy (RRT), duration of MV, length of ICU stay and 28-day mortality between the two group were compared. Results:① A total of 94 patients were enrolled, including 51 in the EGDT group and 43 in the PiCCO group. There was no significant difference in gender, age, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, sequential organ failure assessment (SOFA) score, procalcitonin (PCT), HR, MAP, CVP, Lac or SCr at ICU admission between the two groups. ② The parameters of hemodynamics, fluid balance, urine volume and SCr were improved with the time of resuscitation in the two groups, and there was no significant difference in HR, MAP or Lac between the two groups. Compared with the EGDT group, the CVP decreased significantly at 24 hours and 48 hours after fluid resuscitation in the PiCCO group (mmHg: 9.1±0.9 vs. 12.0±1.3 at 24 hours, 8.0±1.0 vs. 10.2±1.3 at 48 hours), the fluid balance significantly decreased (mL: 2 929.8±936.3 vs. 3 898.4±923.5 at 24 hours, 3 143.5±1 325.4 vs. 4 843.8±1 326.7 at 48 hours), and the condition of urine volume and SCr were better in the PiCCO group [urine volume (mL·kg -1·h -1): 1.02±0.21 vs. 0.79±0.14 at 24 hours, 1.28±0.18 vs. 0.94±0.22 at 48 hours; SCr (μmol/L): 145.7±37.6 vs. 164.3±46.4 at 24 hours, 128.4±33.6 vs. 143.5±37.7 at 48 hours), with significant differences (all P < 0.05). ③ Compared with the EGDT group, the rate of RRT in the PiCCO group was lower [11.6% (5/43) vs. 17.6% (9/51)], the duration of MV and the length of ICU stay were shorter [duration of MV (days): 4.64±1.31 vs. 6.50±2.19, length of ICU stay (days): 10.35±3.50 vs. 14.50±5.78), with significant differences (all P < 0.05). There was no significant difference in the 28-day mortality between the PiCCO group and EGDT group [14.0% (6/43) vs. 15.7% (8/51), P > 0.05]. Conclusion:Fluid resuscitation strategy guided by PiCCO in septic shock patients with AKI can reduce the amount of fluid load, improve renal function, shorten the MV duration and length of ICU stay, and shows clinical significance.

Objective To study the clinical value of double balloon endoscopy (DBE) in diagnosis and treatment of intestinal diseases. Methods Clinical data of 344 patients suspected of small bowel lesions from March 2006 to April 2014 was retrospectively analyzed. All the patients clinical manifestations including varying degrees of abdominal pain, bloating, nause and vomiting, diarrhea. The 344 patients underwent DBE for 397 times. Results The incidence of adverse reactions in checking was only 4.53%, and there was no complications occurred after checking. Positive results shown in 214 cases by DBE, the other 130 patients shown negative results. While 28 of the other 130 patients were found small bowel lesions by capsule endoscopy and CT scans, the remaining 102 cases had no obvious abnormalities. The misdiagnosis rate of DBE was 8.13%, and mainly focuses on polyps and vascular malformation. 242 of the 344 patients were diagnosed with intestinal diseases, and mostly of them were tumor, ulcers and polyps. The detect rate of DBE examination for tumor and ulcer diseases was greater than that of small intestinal polyposis, the difference was statistically significant (P < 0.05). Conclusion DBE in diagnosis and treatment of intestinal diseases is a relatively safe and effective way. It is worthy being popularized and applied.

Background and objective Our previous study found that nicotine could induce lung cancer cell epithelial-mesenchymal transition (EMT). The aim of this study is to explore the relationship between nicotine-induced EMT and lung cancer invasion and metastasis. Methods Real-time PCR and Western blot were used to detect the expression changes of EMT-related markers, E-cadherin and Vimentin, in A549 lung cancer cells treated with nicotine;hTe transposition ofβ-catenin protein expression was determined by immunolfuorescence;Scratch test and Transwell invasion assay were used to detect the effects of nicotine on lung cancer cell migration and invasion. Results Nicotine can signiifcantly down-regulate the expressional level of E-cadherin mRNA and protein of A549 cells in a manner of dose and time-dependent (P<0.01, P<0.01);Nicotine can signiifcantly up-regulate the expressional level of Vimentin mRNA and protein of A549 cells in a manner of dose and time-dependent (P<0.01, P<0.01);Immunolfuorescence results showed thatβ-catenin protein was signiifcantly transfered to nucleus;Scratch test and Transwell assay showed that Nicotine could remarkably increase the migration and invasion poten-tial of lung cancer cells (P<0.01, P<0.01). Conclusion Nicotine can induce cancer cells EMT, and promote the invasion and metastasis ability of lung cancer cells.

PAC1 is the neuropeptide pituitary adenylate cyclase activating polypeptide (PACAP) preferring receptor, which belongs to class B G protein-coupled receptors (GPCR) family. PAC1 mediates the most effects of PACAP as neurotransmitter, neuroregulator and neuroprotectant, while its high expression has close relationship with some physiological and pathological processes such as nerve-injury and tumor. To further understand the function of PAC1, a cell line that expressed inducible PAC1 was constructed to achieve Doxycycline (Dox) dependent expression of PAC1 in CHO (Chinese hamster ovary) cell using the improved Tet (tetracycline)-on Advanced System. First, the PAC1-EYFP fusion gene composed of PAC1 gene and gene encoding EYFP (enhanced yellow fluorescent protein) was sub-cloned to the tetracycline response element pTRE-Tight vector to construct the recombinant vector pEYFP-PAC1-EYFP by double enzyme digestion. Second, the tetracycline regulation components pTet-On advanced vector and the response element pTRE-PAC1-EYFP vector were both introduced into CHO cells successively and the positive clones were screened with G418 and hygromycin respectively. Third, the controlled expression of PAC1-EYFP in CHO was induced by tetracycline analogues Dox in different concentrations and the different levels of receptor PAC1-EYFP were detected. The results of fluorescence analysis and western blotting show that the cell strain with Dox dependent expression of PAC1-EYFP named PAC1-Tet-CHO was obtained. Moreover, in PAC1-Tet-CHO cells the expression of PAC1-EYFP was induced by Dox in a dose-dependent manner. The inducible expression of PAC1 still was stable after sub-culturing for more than 10 passages. It was also found by MTT assay that the higher expression level of PAC1 endowed the cells with higher proliferative viabilities. The construction of controlled expression system of PAC1 will lay a foundation for the further research on PAC1 profiles.
Animals ; Blotting, Western ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Genetic Vectors ; Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I ; biosynthesis