Glycyrrhizin acid and licorice flavonoids are the component of Glycyrrhiza uralensis Fisch root that has been used for various medicinal purposes in traditional oriental medicine for thousands of years. Macrophages as a principal component of immune system play an important role in the initiation, modulation and final activation of immune response against pathogens. In the present study, glycyrrhizin acid and licorice flavonoids was investigated the anti-inflammatory effect on lipopolysaccharide (LPS)-induced macrophage cell line of RAW264.7. Well-grown RAW264.7 cells were collected and randomly divided into the blank control group, the LPS(1 mg x L(-1)) group, the dexamethasone (5 mg x L(-1)) with LPS group, the glycyrrhizin acid (400, 80, 16 mg x L(-1)) with LPS group and the licorice flavonoids (200, 40, 8 mg x L(-1)) with LPS group. RAW264.7 cells were cultured in 24-well plates, pre-incubated for 4 h with different concentrations of dexamethasone, glycyrrhizin acid, or licorice flavonoids. Then cells were stimulated for 20 h with LPS. The supernatant of culture medium was collected from each well and determinated the concentrations of cytokines by means of BioPlex mouse cytokines assay. Compared with the control group, the LPS group could significantly induced relatively high levels of granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor( GM-CSF), macrophage inflammatory protein-1 alpha (MIP-1α), macrophage inflammatory protein-1 beta (MIP-1β), regulated upon activation normal T cell expressed and secreted factor (RANTES), tumor necrosis factor alpha ( TNF-α), monocyte chemotactic protein 1 (MCP-1), chemokine (C-X-C motif) ligand 1 (KC), eotaxin, interleukin(IL)-1α, IL-1β, IL-3, IL-4, IL-5, IL-6, IL-10, IL-12 (p40), IL-12 (p70), IL-13, and IL-17 secretion (P < 0.05). The glycyrrhizin acid significantly inhibited IL-1β, IL-3, IL-5, IL-6, IL-10, IL-12 (p40), IL-12 (p70), IL-13, Eotaxin and TNF-α secreted by LPS-stimulated RAW264.7 cells (P < 0.05). The expression levels of IL-6 and Eotaxin were observably decreased in the licorice flavonoids with LPS group (P < 0.05). The data presented here suggested that the glycyrrhizin acid and licorice flavonoids modulate various cytokines secreted by macrophages and were important anti-inflammatory constituent of Licorice.
Animals ; Cell Line ; Cytokines ; genetics ; immunology ; Flavonoids ; pharmacology ; Glycyrrhiza ; chemistry ; Glycyrrhizic Acid ; pharmacology ; Lipopolysaccharides ; immunology ; Macrophages ; drug effects ; immunology ; Mice

To study the anti-inflammatory activity of glycyrrhizin acid, ligustrazine and puerarin. In the study, the liquichip-based high-throughput synchronous detection technique for 23 inflammatory factors, uniform design, comprehensive weight method were adopted to study the effect of different combined administration of glycyrrhizin acid, ligustrazine and puerarin in inhibiting the expression of lipopolysaccharide (LPS)-induced RAW264. 7 cells and multiple inflammatory cytokines. In the study, the uniform design table U₉ (9³) was adopted to design doses of glycyrrhizin acid, ligustrazine and puerarin. The liquichip technique was used to detect the effect of different combined administration of glycyrrhizin acid, ligustrazine and puerarin on the 23 cytokines expressed in LPS-induced mouse macrophage RAW264. 7 inflammation model. The traditional Chinese medicine component optimization software and the improved least angle regression algorithm were used to analyze the dose-effect relationship among the three components and the cytokine inhibition rate and produce the regression equation. The comprehensive weight method was applied to get the optimal dose ratio of glycyrrhizic acid, ligustrazine and puerarin with highest efficacy of 25:2:13 and verify the optimal dose ratio. The verification results were consistent with the prediction trend, indicating the accuracy of the mathematical model for predicting the experiment. The experimental results showed the multi-target and multi-level efficacies of glycyrrhizic acid, ligustrazine and puerarin and the high anti-inflammatory activity of their combined administration, which provides powerful basis for subsequent drug development.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Cytokines ; Glycyrrhizic Acid ; pharmacology ; Isoflavones ; pharmacology ; Lipopolysaccharides ; immunology ; Macrophages ; drug effects ; immunology ; Mice ; NF-kappa B ; genetics ; immunology ; Pyrazines ; pharmacology ; RAW 264.7 Cells

OBJECTIVE:To investigate the method for rapid and effective isolating of calcium-tolerant cardiomyocytes(CTC) from rats. METHODS: CTC were obtained by retrograde perfusion using modified Langendorff heart perfusion apparatus. After incubation of cardiomyocytes in KB solution, the ion currents were recorded with patch-clamp technique in whole-cell mode. RESULTS: Of the whole isolated cells, 70% were survival. Of which, 80% were CTC. A typical outward potassium current was recorded. CONCLUSIONS: A large number of CTC with normal electrophysiological characteristics can be obtained by mastering the digestion time based on variation of perfusion pressure and keeping all the factors in cell isolation under control.

Objective In the present study we used real-time quantitative RT-PCR to measure the relative expression levels of CGRP,CRLR,RDC-1,RAMP1-3 in periaqueductal gray(PAG)of intact rats. Methods With ?-actin as an endogenous control,write down the C T value and calculate the relative amount of every sample against its endogenous control. Results The results showed the specific amplification of CGRP,CRLR,RAMP1-2 in PAG of rats.Furthermore,the expression of CGRP and RAMP1 is the highest,that of CRLR is higher and the RAMP2 is low;whereas no band is detected in RDC-1 and RAMP3. Conclusion These findings suggest the specific expression of CGRP,CRLR,RAMP1 and RAMP2 mRNA and indicate their functional role in the modulation in PAG of intact rats.

OBJECTIVETo find out the rate of comorbidities of depression, anxiety disorder and attention deficit hyperactivity disorder (ADHD) symptoms in children with epilepsy and to analyze the relevant affecting factors and impacts on quality of life.

METHODTotally 142 children with various types of epilepsy underwent neuropsychological assessment with the Depression Self-rating Scale for Children, the Screen for Child Anxiety Related Emotional Disorders and the ADHD Rating Scale-IV, an 18-item parent-rated questionnaire based on the diagnostic criteria for ADHD, the quality of life was measured in 100 cases on antiepileptic medications by the Quality of Life in Epilepsy Inventory (QOLIE-31). The comorbidity rates were calculated using t-test, chi(2) test and multiple logistic analysis, the variables associated with psychiatric comorbidities were determined, and the impact on quality of life was analyzed.

RESULT(1) The total rate of emotional and behavioral comorbidities was 57.7% (82/142), the frequency of depressive disorder, anxiety disorder and ADHD was 14.8%, 44.4% and 17.6%, respectively. The suicidal ideation occasionally occurred in 5.6% of the cases and 0.7% of cases often had the ideation, but no suicidal action was found in any case. (2) Risk factors for the emotional and behavioral disorders: multiple logistic analysis indicated that age, gender and epilepsy illness-related variables were not relative to the comorbidities, P > 0.05, there were interactions among the disorders. (3) The impact on the quality of life: The emotional and behavioral conditions were associated with the low quality of life, which was significantly lower in epileptic children with co-morbid disorder compared to non-comorbidities epilepsy group. Especially negative impact on the total score of quality of life and four sub-items such as overall quality, emotional well-being, cognitive and social function, P < 0.001. There were also significant differences between the two groups in the other three sub-items including fear for seizure attack, energy/fatigue and medication effects (P < 0.05).

CONCLUSIONSThe frequency of emotional and behavioral disorders including depress disorder, anxiety disorder and ADHD was considerably high in children with epilepsy. Age, gender and epilepsy illness-related variables are not associated with the emotional and behavioral comorbidities, which interfere with each other. Emotional and behavioral disorder is one of the negative factors to the quality of life in epileptic patients. Neuropsychological assessment and treatment are important for improvement of the quality of life in children with epilepsy.

Attention Deficit Disorder with Hyperactivity ; epidemiology ; Child ; Child Behavior Disorders ; epidemiology ; pathology ; Comorbidity ; Emotions ; Epilepsy ; epidemiology ; psychology ; Female ; Humans ; Quality of Life ; Surveys and Questionnaires

UNLABELLEDStudy on the effect of a community-based early education and service program regarding intelligence

OBJECTIVETo assess the effect on a community-based early education and service programs regarding the development of infants' intelligence.

METHODSA community-based intervention study was carried out among 359 infants and their families. Base-line survey were carried out when infants reached their one month, where after infants and their families in the intervention group received instructions and services focused on baby fostering and intelligence development. When the infants reached their six-months and twelve-months of age, their families were informed to complete the follow-up surveys, using both questionnaire investigation and testing the infants' intelligence quotients. The infants' intelligence quotients were measured by Development Screening Test for children under six. By comparing intelligence quotients of infants in two study groups in the follow-up surveys, this paper evaluated the impacts of community intervention on the infants' intelligence development.

RESULTSDuring two follow-ups, no statistical difference had been detected between the two groups of infants in term of gender or delivery process. Baseline data showed that infants' mental index (MI) scored 98.26 in the intervention group and 101.79 in the control one, and development quotient (DQ) scored 94.50 and 99.36 in respective groups. Infants' MI score increased 6.07 and 8.86 at the six-month and twelve-month follow-up periods respectively in the intervention group compared during the baseline, higher than the MI increments of the control group at the two follow-up periods (-2.46 and 1.05 respectively). DQ score of infants in the intervention group increased 12.94 and 11.24 respectively in the two follow-up surveys, which were also higher than increments in the control group (-0.18 and 0.34). The group x time effect(interaction effect) of MI and DQ in six-month and twelve-month follow-ups were both significantly higher than that of the baseline level.

CONCLUSIONThe community-based early education and service programs could effectively improve the infants' intelligence.

Child Development ; Community Health Services ; Female ; Humans ; Infant ; Intelligence ; Male

Objective To investigate the prevalence and associated risk factors of hypertension in Songnan community so as to provide evidence for the early prevention of hypertension.Methods Cluster sampling was employed,and questionnaire survey was performed in 10 185 residents aged above 40 years in Songnan community of Baoshan District.The questionnaires involved with hypertension and associated risk factors.Results The prevalence of hypertension was 49.9% in Songnan community of Baoshan District,and that of men was significantly higher than that of women(53.7% vs 47.3%)(P

BACKGROUNDProneurotrophins such as the precursor of nerve growth factor (proNGF) and the precursor of brain-derived neurotrophic factor (proBDNF) interacted with sortilin and p75(NTR) to form a complex capable of activating an apoptotic signaling. We found that the expression of p75(NTR) and sortilin was increased in ischemic retina induced by elevated intraocular pressure (IOP), but the protein expression changes of proNGF and proBDNF in the same situation were not clear. This study aimed to ascertain the protein expression changes of proNGF and proBDNF in ischemic retina induced by elevated IOP.

METHODSExpression of proBDNF and proNGF was examined by double-labeling immunochemistry in normal rat retina, examined using Western blotting and analyzed using statistical methods in ischemic retina induced by elevated IOP.

RESULTSImmunocytochemistry showed that the proBDNF expressed in the ganglion cell layer (GCL) while the proNGF primarily existed in both the nerve fiber layers (NFL) and large ganglion cell bodies of normal rat retina. Western blotting analysis demonstrated that the molecule weights of 28 kD (proBDNF)/25 kD (proNGF) band were increased significantly (P < 0.05) at days 3, 5 and 7 after retinal elevated-IOP-induced ischemia.

CONCLUSIONProBDNF expressed in the GCL and proNGF primarily presented in NFL and large ganglion cell bodies of normal rat retina, the protein expression forms of 28 kD proBDNF and 25 kD proNGF increased in ischemic retina induced by elevated IOP.

Animals ; Blotting, Western ; Brain-Derived Neurotrophic Factor ; analysis ; Immunohistochemistry ; Intraocular Pressure ; physiology ; Ischemia ; metabolism ; Male ; Nerve Growth Factor ; analysis ; Protein Precursors ; analysis ; Rats ; Rats, Wistar ; Retinal Diseases ; metabolism

Tissue engineering, as a new technology, provides a new avenue for the reconstruction of circumferential tracheal defects, which has always been a tremendous challenge for surgeons around the world. Recently, technologies such as decellularization, 3-dimensional printing, electrospinning and cell sheet have significantly enhanced the chondrification. Implantation of epithelial cells or transplantation of epithelial cell sheets also has accelerated the process of epithelialization. And pedicle muscle flap proved to be a reliable strategy for vascularization of tissue-engineered trachea. But it is still a huge challenge to achieve circumferential tracheal functional reconstruction. The key difficulty lies in how to simultaneously realize the functional regeneration of cartilage, blood vessels and epithelial tissues of tissue-engineered trachea. Therefore, how to integrate the above schemes and finally realize segmental tracheal reconstruction needs further research. This article reviews the research progress of repairing circumferential tracheal defects based on tissue engineering technology.
Printing, Three-Dimensional ; Reconstructive Surgical Procedures ; Tissue Engineering ; Tissue Scaffolds ; Trachea/surgery*

Gene editing is a kind of technologies that makes precise modification to the genome. It can be used to knock out/in and replace the specific DNA fragment, and make accurate gene editing on the genome level. The essence of the technique is the DNA sequence change with use of non homologous end link repair and homologous recombination repair, combined with specific DNA target recognition and endonuclease.This technology has wide range of development prospects and high application value in terms of scientific research, agriculture, medical treatment and other fields. In the field of gene therapy, gene editing technology has achieved cross-time success in cancers such as leukemia, genetic disorders such as hemophilia, thalassemia, multiple muscle nutritional disorders and retrovirus associated infectious diseases such as AIDS and other diseases. The preparation work for new experimental methods and animal models combined with gene editing technology is under rapid development and improvement. Laboratories around the world have also applied gene editing technique in prevention of malaria, organ transplantation, biological pharmaceuticals, agricultural breeding improvement, resurrection of extinct species, and other research areas. This paper summarizes the application and development status of gene editing technique in the above fields, and also preliminarily explores the potential application prospect of the technology in the field of traditional Chinese medicine, and discusses the present controversy and thoughts.