Simplification of genotyping techniques of the ABO blood type experiment and exploration of population genetics.

The ABO blood type is one of the most common and widely used genetic traits in humans. Three glycosyltransferase-encoding gene alleles, IA, IB and i, produce three red blood cell surface antigens, by which the ABO blood type is classified. By using the ABO blood type experiment as an ideal case for genetics teaching, we can easily introduce to the students several genetic concepts, including multiple alleles, gene interaction, single nucleotide polymorphism (SNP) and gene evolution. Herein we have innovated and integrated our ABO blood type genetics experiments. First, in the section of Molecular Genetics, a new method of ABO blood genotyping was established: specific primers based on SNP sites were designed to distinguish three alleles through quantitative real-time PCR. Next, the experimental teaching method of Gene Evolution was innovated in the Population Genetics section: a gene-evolution software was developed to simulate the evolutionary tendency of the ABO genotype encoding alleles under diverse conditions. Our reform aims to extend the contents of genetics experiments, to provide additional teaching approaches, and to improve the learning efficiency of our students eventually.

TRIM65 Suppresses oxLDL-induced Endothelial Inflammation by Interaction with VCAM-1 in Atherogenesis.

BACKGROUND AND OBJECTIVE: Endothelial cell activation, characterized by increased levels of vascular cell adhesion molecule 1 (VCAM-1), plays a crucial role in the development of atherosclerosis (AS). Therefore, inhibition of VCAM-1-mediated inflammatory response is of great significance in the prevention and treatment of AS. The tripartite motif (TRIM) protein-TRIM65 is involved in the regulation of cancer development, antivirals and inflammation. We aimed to study the functions of TRIM65 in regulating endothelial inflammation by interacting with VCAM-1 in atherogenesis. METHODS AND RESULTS: In vitro, we report that human umbilical vein endothelial cells (HUVECs) treated with oxidized low-density lipoprotein (oxLDL) significantly upregulate the expression of TRIM65 in a time- and dose-dependent manner. Overexpression of TRIM65 reduces oxLDL-triggered VCAM-1 protein expression, decreases monocyte adhesion to HUVECs and inhibits the production of the inflammatory cytokines IL-1ß, IL-6, IL-8, and TNF-α as well as endothelial oxLDL transcytosis. In contrast, siRNA-mediated knockdown of TRIM65 promotes the expression of VCAM-1, resulting in increased adhesion of monocytes and the release of the inflammatory cytokines IL-1ß, IL-6, IL-8, and TNF-α and enhances endothelial oxLDL transcytosis. In vivo, we measured the high expression of TRIM65 in ApoE-/- mouse aortic plaques compared to C57BL/6J mouse aortic plaques. Then, we examined whether the blood levels of VCAM-1 were higher in TRIM65 knockout ApoE-/- mice than in control mice induced by a Western diet. Furthermore, Western blot results showed that the protein expression of VCAM-1 was markedly enhanced in TRIM65 knockout ApoE-/- mouse aortic tissues compared to that of the controls. Immunofluorescence staining revealed that the expression of VCAM-1 was significantly increased in atherosclerotic plaques of TRIM65-/-/ApoE-/- aortic vessels compared to ApoE-/- controls. Mechanistically, TRIM65 specifically interacts with VCAM-1 and targets it for K48-linked ubiquitination. CONCLUSION: Our studies indicate that TRIM65 attenuates the endothelial inflammatory response by targeting VCAM-1 for ubiquitination and provides a potential therapeutic target for the inhibition of endothelial inflammation in AS.

[Research advances in the limbic system involved in acupuncture treatment of chronic pain].

Pain, especially chronic pain, can cause multiple changes including sensory-discriminative, emotional-affective, and cognitive-behavior changes and thus greatly affects patients' physical and mental health and quality of life. Therefore, multi-dimensional regulation of paralgesia, cognitive impairment, and negative emotion in patients with chronic pain has become a hot spot in recent studies. The brain regions in the limbic system are involved in the formation and expression of "pain sensation-emotion-cognition". Existing evidence suggests that acupuncture has a multi-dimensional comprehensive regulatory effect on chronic pain, and the brain regions in the limbic system also mediate the analgesic effect of acupuncture. However, further studies are still needed to explore the role and mechanism of action of these brain regions in the multi-dimensional regulation of chronic pain by acupuncture. This article reviews the research advances in the neural mechanism of the limbic system in chronic pain and the role of the limbic system in mediating acupuncture analgesia and mainly elaborates on the mechanism of action of the brain regions in the limbic system in the multi-dimensional regulation of chronic pain.

[Electroacupuncture Relieves Pain by Down-regulating Expression of Hippocampal High Mobility Group Protein 1 and Contents of TNF-α and IL-1 ß in Rats with Chronic Neuropathic Pain].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on expression of high mobility group protein 1 (HMGB 1) and related downstream effectors of proinflammatory cytokines in the hippocampus in chronic neuropathic pain rats, so as to investigate its mechanism underlying neuropathic pain relief. METHODS: Male SD rats were randomized into sham, model, and EA groups, with 12 rats in each group. The neuropathic pain model was established by ligature of the left sciatic nerve to induce chronic constriction injury (CCI). EA was applied to bilateral "Zusanli"(ST 36) and "Yanglingquan"(GB 34) for 30 min, once daily for 7 days. The mechanical withdrawal threshold (WMT) was detected using an electronic von Frey anesthesiometer. The expression level of HMGB 1 in the hippocampus was determined using quantitative RT-PCR and Western blot, separately, and the contents of hippocampal TNF-α and IL-1 ß were detected by ELISA. RESULTS: Compared with the sham group, the MWT values were markedly decreased on day 7, 10 and 14 after modeling in the model group (P<0.001). On day 10 and 14 after modeling, the MWT values were significantly up-regulated in the EA group relevant to those of the model group (P<0.05, P<0.01). The expression levels of HMGB1 mRNA and protein, and the contents of hippocampal TNF-α and IL-1 ß were markedly increased in the model group relevant to the sham group (P<0.001), and significantly down-regulated in the EA group relevant to the model group (P<0.001, P<0.01, P<0.05). CONCLUSION: EA stimulation of ST 36-GB 34 can relieve pain in chronic neuropathic pain rats, which may be related to its actions in down-regulating the levels of HMGB 1 and its downstream proinflammatory cytokines TNF-α and IL-1 ß in the hippocampus.

[Down-regulation of HDAC6 Expression Promotes Apoptosis of Human Leukemia K562 Cells].

OBJECTIVE: To study the promoting-apoptosis effect of HDAC6 on the human leukemia cells and its mechanism. METHODS: The siRNA interference technology was used to inhibit the expression of HDAC6 gene, the RT-PCR and Western blot were used to detect the expression of HDAC6 and related signal pathway proteins respectively, the flow cytometry and Hoechest staining were used to detect the apoptosis and morphology changes of K562 cells. RESULTS: Compared with the periphal blood monocyte and bone marrow stromal cells of healthy volunteers, the expression level of HDAC6 in leukemia cell lines was up-regulated significantly(P<0.05); the flow cytometry and Hoechest staining showed that after interference of HDAC6 gene, the apoptosis of K562 cells increased, moreover the cell morphology was changed; the Western blot detection showed that the interfering HDAC6 increased BAX/BCL-2 ratio and cleaved caspase 3 expression, and activated the MAPK, ATK, ERK signaling pathway. CONCLUSION: The interferance of HDAC6 can promote the K562 cell apoptosis, its mechanism may relate with activation of MAPK signaling pathway.

[Research on Acupuncture in Regulating the Inflammatory Reaction in Anti-ischemia-reperfusion Injury].

The inflammatory response during ischemia-reperfusion is one of principal reasons for secondary organ injury. Therefore, the inhibition of inflammation has become a research hot spot. The current research showed that acupuncture can not only directly inhibit the infiltration of inflammatory cells and regulate the expression of inflammatory cytokines, but is also involved in the overall regulation of the inflammatory signaling pathway and inhibition of the inflammatory response, thereby effectively improving ischemia-reperfusion injury. Here, we review research on the mechanism of acupuncture in ischemia-reperfusion injury via regulation of the inflammatory response.