Objective To observe the effects of low-frequency pulsed electromagnetic fields (LFPEMFs) on microcirculation angiogenesis in the hindlimbs of diabetic rats with acute ischemia. Methods Models of acute hindlimb ischemia were established in 60 male Sprague-Dawley diabetic rats. The diabetes model was established using 60 mg/kg intraperitoneal injections of streptozotocin (STZ). Fasting blood glucose levels were greater than 300 mg/dL. The rats were randomly divided into experimental and control groups. The rats in the experimental group were exposed to low-frequency pulsed electromagnetic fields for 2 hours each day, while the control group was not given any treatment. Laser-Doppler perfusion was used to measure blood flow in the ischemic hindlimb on days 0,7, 14 and 28 after the operation. The immunofluorescence of rat endothelial cell antigen-1 ( RECA-1) was used to evaluate the changes in angiogenesis. The levels of vascular endothelial growth factor (VEGF) and fibroblast growth factor-2 (FGF-2) were determined by both Western blotting and ELISA, and VEGFR2 and FGFR1 levels in the ischemic skeletal muscle were determined by Western blotting on days 7, 14 and 28 after the operation. Results The average perfusion ratio was significantly greater in the experimental group at days 14 and 28 compared with the control group. RECA-1 density in the tissues had increased significantly in the experimental group at the 14th and 28th day. The same was observed for FGF-2 and its receptor, but there was no significant difference for VEGF or its receptor in either group. Conclusions LFPMEFs can promote angiogenesis in acute hindlimb ischemia of diabetic rats by up-regulating FGF-2. This suggests that LFPMEFs may be useful for preventing and treating lower limb ischemia in diabetic humans.

Objective To examine the association of arteriovenous fistula (AVF) blood flow (Qa) dynamics with inflammation state and its effect on cardiovascular diseases (CVD) in maintenance hemodialysis (MHD) patients.Methods Thirty MHD patients with AVF and twelve healthy people were enrolled in the study.Qa and cardiac output (CO) were measured by Transonic Hemodialysis Monitor HD 02.In MHD patients,pre-dialysis blood samples were taken before Qa monitoring.High-sensitivity C-reactive protein (hsCRP) was measured by immunoturbidimetry (Kyoma,Japan).Inflammatory factors IL-2,IL-6,IL-10,TNF were measured by Cytometric Bead Array (BDTM).Cardiovascular diseases morbidity was monitored prospectively within nineteen months follow-up period.Results There were no significant differences in age and sex between MHD patients and healthy people.The serum IL-6,IL-10,TNF and hsCRP were significantly higher in MHD patients than those in healthy controls [2.38 (1.86-4.69) vs 1.14 (0.27-1.18) ng/L,P＜0.01; 1.47 (1.19-2.10) vs 1.04 (0.00-1.23) ng/L,P＜0.01; 1.33 (1.05-1.56) vs 0.54 (0.00-1.24) ng/L,P＜0.05; 4.90 (1.58-7.45) vs 1.50 (0.63-1.90) mg/L,P=0.01].During the follow-up period,6 patients (20.0％) developed at least one episode of cardiovascular event.Qa,serum IL-6 and hsCRP levels were significantly higher in patients with CVD as compared to those without CVD [(1120±192) vs (893±189) ml/min,P＜0.05; 4.86 (2.96-7.85) vs 2.20 (1.80-3.10) ng/L,P＜ 0.01;11.75 (3.83-31.53) vs 4.45 (1.05-6.68) mg/L,P＜0.05].Binary Logistic regression analysis demonstrated that serum IL-6 was an independent and stronger risk factor for CVD morbidity [HR=1.943,95％CI (1.110-3.402),P=0.02].Spearman rank correlation analysis and liner regression analysis showed that Qa was positively correlated with serum IL-6 (β=0.492,P＜0.01).Path analysis suggested that Qa contributed to CVD mortality via the increase of serum IL-6.Conclusions AVF blood flow monitoring is important for MHD patients.IL-6 is an independent risk factor of CVD in MHD patients.AVF blood flow increases cardiovascular diseases morbidity in MHD patients via its promotion of IL-6 production.

Due to the complexity of bioactive ingredients in biological samples,the screening of target proteins is a complex process.Herein,a feasible strategy for directing protein immobilization on silica magnetic beads for ligand fishing based on SpyTag/SpyCatcher(ST/SC)-mediated anchoring is presented.Carboxyl functional groups on the surface of silica-coated magnetic beads(SMBs)were coupled with SC using the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysulfosuccinimide method,named SC-SMBs.The green fluorescent protein(GFP),as the capturing protein model,was ST-labeled and anchored at a specific orientation onto the surface of SC-SMBs directly from relevant cell lysates via ST/SC self-ligation.The characteristics of the SC-SMBs were studied via electron microscopy,energy dispersive spectroscopy,and Fourier transform infrared spectroscopy.The spontaneity and site-specificity of this unique reaction were confirmed via electrophoresis and fluorescence analyses.Although the alkaline stability of ST-GFP-ligated SC-SMBs was not ideal,the formed isopeptide bond was unbreakable under acidic conditions(0.05 M glycine-HCl buffer,pH 1-6)for 2 h,under 20％ethanol solution within 7 days,and at most temperatures.We,therefore,present a simple and universal strategy for the preparation of diverse protein-functionalized SMBs for ligand fishing,prompting its usage on drug screening and target finding.

Objective To observe the effect of external recombinant human basic fibroblast growth factor (rh-bFGF) on the skin and muscles.Methods 40 BALB/c mice of 6 weeks were randomly divided into skin and muscle groups.The skin group was randomly divided into group A and group B.The skin group was injected with 100U rh-bFGF and the same dose of saline in the chin.The muscle groups were randomly divided into C,D,E,and F,and the muscle group was injected into the left calf muscle in order of 200,400,800U rh-bFGF and the same dose of normal saline.The materal was taken at week 8.HE staining was used to observe the submental tissue and muscle fiber morphology.Two groups of muscle fibers,collagen and fibrous tissue were detected by trichrome stain.Immunohistochemistry was used to determine the blood vessel density of the chin and gastrocnemius.Results In week 8,10 mice in group A had a mass in the lower jaw,and the gastrocnemius of the groups C and D showed hypertrophy.Group A showed the thickness of epidermis,dermal papillary layer and mucosa,hair follicle and blood vessel quantity,glandular cavity,collagen and fiber content were sigificantly greater than that of group B (P＜0.05).In the groups C,D and E,the gastrocnemius muscle was significantly thicker than that of group F,and the thickening of group D was the most obvious (P＜0.05),and the contents of collagen and fiber in the groups C,D and E were significantly higher than that in group F.Conclusions Subcutaneous injection of rh-bFGF leads to submental skin thickening,vascular hyperplasia,increased diameter of hair follicle and higher collagen and fiber contents;intramuscular injection of it may induce the muscular hypertrophy and vascular proliferation.

A quantitative analytical method for multi-components with a single-marker (QAMS) was established for simultaneous determination of neochlorogenic acid, chlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Artemisia capillaris Thunb standard decoction. The separation was performed on a Waters CORTECS T3 column (2.1 mm × 100 mm, 2.7 μm), with the mobile phase consisting of 0.05% phosphate acid solution-acetonitrile for gradient elution. The column temperature was 30 ℃, and flow rate was 0.5 mL·min^-1. Using chlorogenic acid as an internal reference, the relative correlation factors of neochlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid were calculated following UHPLC, as 0.928 0, 0.546 2, 1.099 8, 0.872 1, 1.086 8, 0.739 2, 1.056 6, respectively. The results were compared with those obtained by the external standard method to verify the feasibility, rationality and repeatability of QAMS method. There was no significant difference in assay results between QAMS and the external standard method. In conclusion, the QAMS method is accurate and feasible, and could be used to determine the content such as neochlorogenic acid, caffeic acid, cryptochlorogenic acid, 1,3-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid in Artemisia capillaris Thunb standard decoction.

Coronavirus disease-2019 (COVID-19) belongs to the epidemic diseases of traditional Chinese medicine (TCM), which is infected by the air with disease and the initial stage of the disease is in upper energizer. TCM holds that the nose is the orifice of the lung and the gateway of the breath. WU Shang-xian, the famous external doctor in Qing dynasty, discussed in Liyue Pianwen that "for the disease in upper energizer, the most effective method is to use the medicine powder via nasal administration and sneeze to disperse". For thousands of years, ancient Chinese medical practitioners had explored and developed the TCM nasal administration method in the struggle against epidemics. Qingkailing is the basic formula for heat-clearing and detoxifying, and researches have clarified its therapeutic effect on upper respiratory tract infections. Therefore, based on TCM nasal administration, this article took Qingkailing as an example to study the feasibility of its nasal preparations for the treatment of COVID-19. On the one hand, it is helpful for the rapid development of Qingkailing nasal preparations for COVID-19. On the other hand, it can broaden the new thinking of TCM in treating epidemic diseases, and give full play to the advantages of TCM in treating epidemic diseases.

In this study, the texture analyzer acupuncture pressure sensor was used to objectively characterize the "herb soaking with exact amount of water" for moistening process of ginseng. The single factor rotation experiment was used to investigate the effects of puncture speed, puncture depth and puncture site on puncture force and work. According to ginseng processing method in Chinese Pharmacopoeia, ginseng medicinal materials with diameters of about 1 cm and 2 cm were selected, and puncture experiments were carried out at the set measurement time to determine the hardness, work and water absorption of the ginseng moistening process. The endpoint threshold for the ginseng softening process was determined and verified. To reflect the actual internal conditions of the ginseng softening process, the puncture depth was preferably 70%, and the puncture speed was 30 mm·min~(-1). In the ginseng moistening process, the softening hardness and the puncture work were in accordance with the first-order kinetic equation y=a×exp(-k×x). The 0 h initial hardness a of 1 cm and 2 cm ginseng herbs were 289.8 N and 1 227 N, and the rate constants K were 0.149 4 N·h~(-1) and 0.100 7 N·h~(-1), respectively. After the ginseng was completely softened, the force required for puncture was 10 N, which can be used as the standard for "drug penetration". At this time, the water absorption rate of ginseng was 70%-100%. The softening time of ginseng with a diameter of 1 cm was about 20-22 h, and the softening time of ginseng with a diameter of 2 cm was about 40-46 h. A needle-type pressure sensor was used to accurately determine the end point of the softening process of ginseng and reduce the loss of active ingredients. The study results provide reference for the softening process kinetics and the process intelligent monitoring of other dried roots and rhizomes.
Drugs, Chinese Herbal ; Panax ; Plant Roots ; Rhizome ; Technology, Pharmaceutical ; Water

Predatory hunting is an important type of innate behavior evolutionarily conserved across the animal kingdom. It is typically composed of a set of sequential actions, including prey search, pursuit, attack, and consumption. This behavior is subject to control by the nervous system. Early studies used toads as a model to probe the neuroethology of hunting, which led to the proposal of a sensory-triggered release mechanism for hunting actions. More recent studies have used genetically-trackable zebrafish and rodents and have made breakthrough discoveries in the neuroethology and neurocircuits underlying this behavior. Here, we review the sophisticated neurocircuitry involved in hunting and summarize the detailed mechanism for the circuitry to encode various aspects of hunting neuroethology, including sensory processing, sensorimotor transformation, motivation, and sequential encoding of hunting actions. We also discuss the overlapping brain circuits for hunting and feeding and point out the limitations of current studies. We propose that hunting is an ideal behavioral paradigm in which to study the neuroethology of motivated behaviors, which may shed new light on epidemic disorders, including binge-eating, obesity, and obsessive-compulsive disorders.
Animals ; Zebrafish ; Hunting ; Predatory Behavior/physiology* ; Neurons/physiology* ; Motivation