Effects of electroacupuncture on rats with cognitive impairment: An iTRAQ-based proteomics analysis / 中西医结合学报

OBJECTIVE@#The study explores the effects of electroacupuncture (EA) at the governing vessel (GV) on proteomic changes in the hippocampus of rats with cognitive impairment.@*METHODS@#Healthy male rats were randomly divided into 3 groups: sham, model and EA. Cognitive impairment was induced by left middle cerebral artery occlusion in the model and EA groups. Rats in the EA group were treated with EA at Shenting (GV24) and Baihui (GV20) for 7 d. Neurological deficit was scored using the Longa scale, the learning and memory ability was detected using the Morris water maze (MWM) test, and the proteomic profiling in the hippocampus was analyzed using protein-labeling technology based on the isobaric tag for relative and absolute quantitation (iTRAQ). The Western blot (WB) analysis was used to detect the proteins and validate the results of iTRAQ.@*RESULTS@#Compared with the model group, the neurological deficit score was significantly reduced, and the escape latency in the MWM test was significantly shortened, while the number of platform crossings increased in the EA group. A total of 2872 proteins were identified by iTRAQ. Differentially expressed proteins (DEPs) were identified between different groups: 92 proteins were upregulated and 103 were downregulated in the model group compared with the sham group, while 142 proteins were upregulated and 126 were downregulated in the EA group compared with the model group. Most of the DEPs were involved in oxidative phosphorylation, glycolipid metabolism and synaptic transmission. Furthermore, we also verified 4 DEPs using WB technology. Although the WB results were not exactly the same as the iTRAQ results, the expression trends of the DEPs were consistent. The upregulation of heat-shock protein β1 (Hspb1) was the highest in the EA group compared to the model group.@*CONCLUSION@#EA can effect proteomic changes in the hippocampus of rats with cognitive impairment. Hspb1 may be involved in the molecular mechanism by which acupuncture improves cognitive impairment.

Active components and mechanism of Jinwugutong Capsules in treatment of osteoporosis: a study based on UPLC-Q-Exactive-MS/MS combined with network pharmacology / 中国中药杂志

UPLC-Q-Exactive-MS/MS and network pharmacology were employed to preliminarily study the active components and mechanism of Jinwugutong Capsules in the treatment of osteoporosis. Firstly, UPLC-Q-Exactive-MS/MS was employed to characterize the chemical components of Jinwugutong Capsules, and network pharmacology was employed to establish the "drug-component-target-pathway-disease" network. The key targets and main active components were thus obtained. Secondly, AutoDock was used for the molecular docking between the main active components and key targets. Finally, the animal model of osteoporosis was established, and the effect of Jinwugutong Capsules on the expression of key targets including RAC-alpha serine/threonine-protein kinase(AKT1), albumin(ALB), and tumor necrosis factor-alpha(TNF-α) was determined by enzyme-linked immunosorbent assay(ELISA). A total of 59 chemical components were identified from Jinwugutong Capsules, among which coryfolin, 8-prenylnaringenin, demethoxycurcumin, isobavachin, and genistein may be the main active components of Jinwugutong Capsules in treating osteoporosis. The topological analysis of the protein-protein interaction(PPI) network revealed 10 core targets such as AKT1, ALB, catenin beta 1(CTNNB1), TNF, and epidermal growth factor receptor(EGFR). The Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment showed that Jinwugutong Capsules mainly exerted the therapeutic effect by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT) signaling pathway, neuroactive ligand-receptor interaction, mitogen-activated protein kinase(MAPK) signaling pathway, Rap1 signaling pathway and so on. Molecular docking showed that the main active components of Jinwugutong Capsules well bound to the key targets. ELISA results showed that Jinwugutong Capsules down-regulated the protein levels of AKT1 and TNF-α and up-regulated the protein level of ALB, which preliminarily verified the reliability of network pharmacology. This study indicates that Jinwugutong Capsules may play a role in the treatment of osteoporosis through multiple components, targets, and pathways, which can provide reference for the further research.

Small molecule proteomics quantifies differences between normal and fibrotic pulmonary extracellular matrices / 中华医学杂志(英文版)

BACKGROUND@#Pulmonary fibrosis is a respiratory disease caused by the proliferation of fibroblasts and accumulation of the extracellular matrix (ECM). It is known that the lung ECM is mainly composed of a three-dimensional fiber mesh filled with various high-molecular-weight proteins. However, the small-molecular-weight proteins in the lung ECM and their differences between normal and fibrotic lung ECM are largely unknown.@*METHODS@#Healthy adult male Sprague-Dawley rats (Rattus norvegicus) weighing about 150 to 200 g were randomly divided into three groups using random number table: A, B, and C and each group contained five rats. The rats in Group A were administered a single intragastric (i.g.) dose of 500 μL of saline as control, and those in Groups B and C were administered a single i.g. dose of paraquat (PQ) dissolved in 500 μL of saline (20 mg/kg). After 2 weeks, the lungs of rats in Group B were harvested for histological observation, preparation of de-cellularized lung scaffolds, and proteomic analysis for small-molecular-weight proteins, and similar procedures were performed on Group C and A after 4 weeks. The differentially expressed small-molecular-weight proteins (DESMPs) between different groups and the subcellular locations were analyzed.@*RESULTS@#Of the 1626 small-molecular-weight proteins identified, 1047 were quantifiable. There were 97 up-regulated and 45 down-regulated proteins in B vs. A, 274 up-regulated and 31 down-regulated proteins in C vs. A, and 237 up-regulated and 28 down-regulated proteins identified in C vs. B. Both the up-regulated and down-regulated proteins in the three comparisons were mainly distributed in single-organism processes and cellular processes within biological process, cell and organelle within cellular component, and binding within molecular function. Further, more up-regulated than down-regulated proteins were identified in most sub-cellular locations. The interactions of DESMPs identified in extracellular location in all comparisons showed that serum albumin (Alb) harbored the highest degree of node (25), followed by prolyl 4-hydroxylase beta polypeptide (12), integrin β1 (10), apolipoprotein A1 (9), and fibrinogen gamma chain (9).@*CONCLUSIONS@#Numerous PQ-induced DESMPs were identified in de-cellularized lungs of rats by high throughput proteomics analysis. The DESMPs between the control and treatment groups showed diversity in molecular functions, biological processes, and pathways. In addition, the interactions of extracellular DESMPs suggested that the extracellular proteins Alb, Itgb1, Apoa1, P4hb, and Fgg in ECM could be potentially used as biomarker candidates for pulmonary fibrosis. These results provided useful information and new insights regarding pulmonary fibrosis.

Effects of fluoride on growth and development, learning and memory, and oxidative stress in serum of offspring rats / 中华地方病学杂志

Objective:To observe the change of growth and development, learning and memory, and oxidative stress in serum of offspring rats with fluorosis, and to explore the mechanism of fluoride on neurobehavioral development of offspring rats.Methods:Seventy-two SD rats (female and male ratio 3 ∶ 1) were fed adaptively for one week. According to their body mass [(80 ± 20) g], they were divided into control group (drinking tap water, containing less than 0.5 mg/L fluoride), low fluorine group (drinking water containing 10.0 mg/L of fluoride), and high fluorine group (drinking water containing 100.0 mg/L of fluoride) with random number table. After six months of feeding, they mated freely and gave birth in each group (24 rats with 18 females and 6 males). The rats in each group continued to be exposed to fluoride after giving birth, and the offspring rats were exposed to fluoride through breast milk feeding until the 28th day after birth. Body and brain weight, growth and development indicators (auricle separation, eyes opening, teeth eruption and hair growth) and neurobehavioral development indicators (cliff avoidance, auditory startle, surface righting and vibrissa positioning) were recorded. On the 28th day after birth, the learning and memory abilities (escape latency) of offspring rats were tested by Morris water maze; blood samples were taken from eyeballs to detect the content of nitric oxide (NO), the activity of nitric oxide synthase (NOS) and inducible nitric oxide synthase (iNOS).Results:On the 21st day and 28th day after birth, the differences of body weight among control group, low fluorine group and high fluorine group [21st day: (54.70 ± 3.02), (52.30 ± 2.58), (51.30 ± 2.71) g, 28th day: (91.70 ± 5.03), (90.40 ± 4.76), (86.00 ± 4.55) g] were statistically significant ( F = 3.96, 3.70, P < 0.05); on the 21st day, the body weight of high fluorine group was lower than that of control group ( P < 0.05); on the 28th day, the body weight of the high fluorine group was lower than those of control group and low fluorine group ( P < 0.05). On the 28th day, the difference of brain weight of control group, low fluorine group and high fluorine group was statistically significant ( F = 6.19, P < 0.05); and the low fluorine group and high fluorine group were lower than that of control group ( P < 0.05). Among the growth development indicators, the difference of time of completing eyes opening in control group, low fluorine group and high fluorine group was statistically significant ( F = 3.64, P < 0.05); and the high fluorine group was higher than that of control group ( P < 0.05). In neurobehavioral development indicators, the differences of time of completing cliff avoidance, surface righting between the control group, low fluorine group and high fluorine group were statistically significant ( F = 8.29, 7.69, P < 0.05); and the time of completing cliff avoidance in high fluorine group was higher than those of control group and low fluorine group ( P < 0.05), the time of completing surface righting was higher than that of control group ( P < 0.05). In Morris water maze, on the 4th day, the escape latencies of low fluorine group and high fluorine group were higher than that of control group ( P < 0.05). The results of oxidative stress in serum showed that there were statistically significant differences in serum NO content, NOS and iNOS activitives between the control group, low fluorine group and high fluorine group ( F = 4.86, 66.48, 70.95, P < 0.05); and the NO content of the high fluoirne group was higher than that of the control group ( P < 0.05), the activities of NOS and iNOS of the high fluoirne group were higher than those of control group and the low fluorine group ( P < 0.05). Conclusion:Excessive fluoride can increase the level of oxidative stress in serum, which may be closely related to the neurobehavioral retardation and the decline of learning and memory ability of offspring rats.

Mechanism and research progress on radiotherapy combined with immunotherapy / 中华放射肿瘤学杂志

In recent years, the incidence rate of malignant tumors has been ever increased. As the persistent advancement of various therapeutic techniques, the therapeutic plans of cancer have been improved. Radiotherapy takes effect mainly by killing the topical tumor cells by radiation. During radiotherapy, the anti-tumor immune response can be induced or enhanced. Appropriate radiotherapy dose and segmentation model combined with certain immunotherapy plays a more and more significant role in the treatment of tumors. In this article, the underlying mechanisms of radiation-enhanced anti-tumor immune response and the current status and research prospects of radiotherapy combined with immunotherapy were reviewed.

Effect of fluoride on expression of phosphorylated NMDAR protein in rat hippocampal neurons / 中华地方病学杂志

Objective To observe the expression of phosphorylated N-methyl-D-aspartate receptor (P-NMDAR) protein in hippocampal neurons of rats with fluorosis and explore the mechanism of neuronal damage caused by fluorosis.Methods Sixteen Sprague-Dawley rats within 24 h after birth were selected,and hippocampus of the brain was isolated after sacrifice.The primary neurons were cultured in vitro.Observation of the cell morphology under an inverted microscope,neurons were identified by immunofluorescence staining.In the 7th day of cultivation,the neurons were divided into control group,low fluoride group,high fluoride group,antagonist group,low fluoride antagonist group and high fluoride antagonist group.The control group was treated with the same volume of medium as the experimental group.The concentration of NaF was 0.2 and 2.0 mmol/L in the low fluoride group and the high fluoride group,respectively.The antagonist group was treated with 10.0 μ mol/L NMDAR antagonist (MK-801).The low fluoride antagonist group and the high fluoride antagonist group were treated with 0.2 mmol/L NaF + 10.0 μmol/L MK-801,2.0 mmol/L NaF + 10.0 μmol/L MK-801,respectively.The culture time was 24 hours.The expression levels of phosphorylated protein (P-NMDAR1,P-NMDAR2A,P-NMDAR2B) of NMDAR subunits were detected by Western blotting.Results Under inverted microscope,the primary cell body of the cultured in vitro for 2-3 days became larger,and many protrusions appeared outward,showing a small spider shape;6-7 days,the cells synaptic long and slender,a network-like interlaced form.Under fluorescent microscope,NeuN positive cells (neurons) with red fluorescence were observed,and the cell purity exceeded 80％.There was no significant difference in the expression of P-NMDAR1 protein between the control group,the low fluoride group,the high fluoride group,the antagonist group,the low fluoride antagonist group and the high fluoride antagonist group (0.44 ± 0.12,0.46 ± 0.06,0.46 ± 0.12,0.56 ± 0.10,0.70 ± 0.12,0.46 ± 0.09,F=2.75,P ＞ 0.05);P-NMDAR2A and P-NMDAR2B protein expressions were statistically significant between the six groups (0.75 ± 0.17,0.74 ± 0.08,1.13 ± 0.27,0.87 ± 0.15,0.67 ± 0.11,0.66 ± 0.09;0.68 ± 0.24,0.66 ± 0.12,1.46 ±0.27,0.74 ± 0.16,0.56 ± 0.13,0.91 ± 0.35,F =3.68,6.11,P ＜ 0.05),and the expressions of P-NMDAR2A and P-NMDAR2B protein in high fluoride group were higher than those in the control group (P ＜ 0.05);the expressions in the high fluoride antagonist group were lower than those in the high fluoride group (P ＜ 0.05).Conclusion Excessive fluoride can increase the expressions of P-NMDAR2A and P-NMDAR2B protein of hippocampal neurons,and co-culture of MK-801 and NaF can antagonize the damage of fluoride to neurons.

Mechanism of nerve injury caused by fluorosis: a review of recent studies / 中华地方病学杂志

Excessive exposure to fluoride in the environment leads to a variety of health hazards,especially in the nervous system damage.Great number of experimental and clinical data have confirmed that excessive intake of fluoride can cause neurotoxicity,and produce a series of symptoms of central nervous system dysfunction.This review summarizes the mechanisms of neurotoxicity induced by fluorosis in recent years,including oxidative stress,neurotransmitters and receptors,central nervous system signaling molecules,neuronal energy metabolism,and glycosylation end products.This paper analyzes the effects of fluorine exposure on nervous system,thus providing a reliable basis for further prevention and treatment of fluorosis.

Efficacy and Safety of Cinacalcet in the Treatment of Hemodialysis Patients with Secondary Hyperparathy-roidism:A Systematic Review / 中国药房

OBJECTIVE:To systematically review the efficacy and safety of cinacalcet in the treatment of hemodialysis pa-tients with secondary hyperparathyroidism,and provide evidence-based reference for the clinical treatment. METHODS:Retrieved from Medline,Cochrane Library,EMBase and CBM,randomized controlled trials(RCT)about cinacalcet in the treatment of he-modialysis patients with secondary hyperparathyroidism (SHPT) were collected. Meta-analysis was performed by using Rev Man 5.3.5 software after data extract and quality evaluation by Cochrane systematic Rev Man 5.3.5. RESULTS:Totally 7 RCTs were en-rolled,involving 1 987 patients. Results of Meta-analysis showed cinacalcet can significantly reduce the rate of surgical parathyroid-ectomy[RR=0.23,95%CI(0.06,0.89),P=0.03],incidence of fracture[RR=0.26,95%CI(0.12,0.60),P=0.002] and increase the incidences of hypocalcemia[RR=9.81,95%CI(3.92,4.59),P<0.001],nausea[RR=1.97,95%CI(1.58,2.46),P<0.001] and vomit-ing[RR=1.91,95%CI(1.50,2.42),P<0.001],while it showed no significant effect on the the incidence of all-cause mortality and cardiovascular death. CONCLUSIONS:The clinical efficacy of cinacalcet in the treatment of hemodialysis patients with secondary hyperparathyroidism is good,but there are common adverse reactions such as nausea and vomiting,hypocalcemia.

Synthesis and in vitro antibacterial activity of 7-(4-alkoxyimino-3-methyl-3-methylaminopiperidin-1-yl)quinolones / 药学学报

To explore new agents of quinolone derivatives with high antibacterial activity, 7-(4-alkoxyimino-3-methyl-3-methylaminopiperidin-1-yl)quinolones were designed and synthesized, and their activity against gram-positive and gram-negative strains was tested in vitro. Sixteen target compounds were obtained. Their structures were established by 1H NMR, HRMS and X-ray crystallographic analysis. Compounds 14k and 14m-14o show good antibacterial activity against the tested five gram-positive strains and five gram-negative strains (MIC: 0.25-16 micromg x mL(-1)), of which the most active compound 14o is 8-fold more potent than levofloxacin against S. pneumoniae (MIC: 4 microg x mL(-1)), and comparable to levofloxacin against S. aureus, S. epidermidis, E. faecalis and E. coli (MIC: 0.25-1 microg x mL(-1)), but generally less potent than gemifloxacin.

Symptomatic characteristics of chronic rhinosinusitis patients and symptom-specific outcomes after endoscopic sinus surgery / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the symptomatic characteristics of chronic rhinosinusitis patients and the report symptom-based outcomes after endoscopic sinus surgery.@*METHOD@#One hundred and nineteen chronic rhinosinusitis patients underwent endoscopic sinus surgery, including 52 patients of chronic rhinosinusitis without nasal polyps and 67 patients of chronic rhinosinusitis with nasal polyps, were enrolled. Patients were asked to evaluate their symptoms before surgery and 12 months after endoscopic sinus surgery using 10 cm visual analog scale measures.@*RESULT@#The most commonly reported symptoms were nasal obstruction, nasal discharge, headache, facial pressure and altered sense of smell. Compared with patients of chronic rhinosinusitis with nasal polyps, patients of chronic rhinosinusitis without nasal polyps reported significantly higher scores of nasal discharge, whereas lower scores of altered sense of smell (P<0.01 for both). The most disturbing symptom was nasal discharge and altered sense of smell for chronic rhinosinusitis patients without and with nasal polyps, respectively. After endoscopic sinus surgery, the scores for all studied symptoms were improved greatly in both chronic rhinosinusitis with and without nasal polyps groups (P<0.01 for all).@*CONCLUSION@#Chronic rhinosinusitis patients with and without nasal polyps present different symptomatic characteristics. Endoscopic sinus surgery can improve patient-based symptoms significantly. Visual analog scale is a simple and powerful tool to evaluate chronic rhinosinusitis symptoms.