Augmentative effect of tetrandrine on pentobarbital hypnosis mediated by 5-HT_(1A) and 5-HT_(2A/2C) receptors in mice

It has been reported that augmentative effect of tetrandrine on pentobarbital hypnosis in mice may be related to serotonergic system. The present study was undertaken to investigate the interaction of tetrandrine and different 5-HT receptors on pentobarbital-induced sleep by using the loss-of-righting reflex method. The results showed that augmentative effect of tetrandrine on pentobarbital hypnosis in mice were potentiated by the p-MPPI （5-HT1A receptor antagonist） （1 mg/kg, i.p.） and ketanserin （5-HT2A/2C receptor antagonist） （1.5 mg/kg, i.p.）, respectively. Pretreatment with either 8-OH-DPAT （5-HT1A receptor agonist） （0.1 mg/kg, s.c.） or DOI （5-HT2A/2C receptor agonist） （0.2 mg/kg, i.p.） significantly decreased pentobarbital-induced sleep time, and tetrandrine （60 mg/kg, i.g.） significantly reversed this effect. These results suggest that both the 5-HTLA and 5-HT2A/2C subfamily may be involved in the potentiating mechanism of tetrandrine＇s effects on pantobarbital hypnosis.

The 5-HT2c receptor gene Cys23Ser polymorphism influences the intravaginal ejaculation latency time in Dutch Caucasian men with lifelong premature ejaculation

It has been postulated that the persistent short intravaginal ejaculation latency time （IELT） of men with lifelong premature ejaculation （LPE） is related to 5-hydroxytryptamine （HT）2c receptor functioning. The aim of this study was to investigate the relationship of Cys23Ser 5-HT2c receptor gene polymorphism and the duration of IELT in men with LPE. Therefore, a prospective study was conducted in 64 Dutch Caucasian men with LPE. Baseline IELT during coitus was assessed by stopwatch over a 1-month period. All men were genotyped for Cys23Ser 5-HT2c receptor gene polymorphism. Allele frequencies and genotypes of Cys and Ser variants of 5-HT2c receptor gene polymorphism were determined. Association between Cys/Cys and Ser/Ser genotypes and the natural logarithm of the IELT in men with LPE were.investigated. As a result, the geometric mean, median and natural mean IELT were 25.2, 27.0, 33.9s, respectively. Of all men, 20.0%, 10.8%, 23.1% and 41.5% ejaculated within 10, 10-20, 20-30 and 30-60s after vaginal penetration. Of the 64 men, the Cys/Cys and Ser/Ser genotype frequency for the Cys23Ser polymorphism of the 5-HT2c receptor gene was 81% and 19%, respectively. The geometric mean IELT of the wildtypes （Cys/Cys） is significantly lower （22.6s; 95% CI 18.3-27.8s） than in male homozygous mutants （Ser/Ser） （40.4s; 95% CI 20.3-80.4s） （P = 0.03）. It is concluded that Cys23Ser 5-HT2c receptor gene polymorphism is associated with the IELT in men with LPE. Men with Cys/Cys genotype have shorter IELTs than men with Ser/Ser genotypes.

Effects of Estradiol on 5-HTsA and 5-HT2c Receptor Immunolabeling in Rat Hippocampus

Steroid hormones participate in the modulation of serotonergic transmission, including the regulation of synthetic and metabolic enzyme production, as well as receptor and transporter activity. The changes of 5-HT5A and 5-HT2c immunolabeling induced by steroids in the hippocampus ofovariectomized rats were studied in this work. Densitometric analysis in rat hippocampi were carried out for adjacent brain coronal immunolabeled sections after treatment with subcutaneous injections of vehicle, estradiol, progesterone or the combination of both steroids in ovariectomized rats. Exposure to estradiol and the combination of estradiol and progesterone significantly reduced the 5-HT5A-like immunosignal in the CA 1 region while progesterone did not induce changes. On the other hand, exposure to the combination of estradiol and progesterone or estradiol alone increased the 5-HT2c immunosignal in the same region. These results indicate that estradiol is involved in the discrete regulation of serotonin receptors 5-HT5A and 5-HT2c in rat hippocampus.

Role of 5-HT2A Receptors in Immunomodulation in Animal Models of Aggressive Behavior

Serotonin 5-HT2A receptors are playing an important role in the pathophysiology of aggressive behaviors and in the control of immune function. In the present study, we analyzed the effects of activation and blockade of 5-HT2A receptors with selective ligands on the immune response formation in animals with aggressive behaviors induced by genetic factors (rats selected for the increased aggressiveness toward human) or by chronic social stress (mice of the CBA/Lac strain engaged in 10 days of social confrontations). Activation of 5-HT2A receptors with DOI at 1.0 mg/kg reduced the immune response level both in aggressive rats and mice compared to the corresponding vehicle-treated groups, while DOI administration did not alter the immune reaction in nonaggressive animals. The blockade of 5-HT2A receptors with ketanserin at 1.0 mg/kg resulted in immunostimulation both in mice of the CBA strain not subjected to social stress (the controls) and in nonaggressive rats selected for elimination of aggressiveness. On the other hand, its administration to CBA mice demonstrating offensive aggression enhanced the immune reaction, while the same dose of ketanserin did not modify the immune response level in rats with genetic predisposition to the increased defensive aggression. Thus, our data suggest that the role of 5-HT2A receptors in immunomodulation depends on the specific type of aggression that may be taking into account in the treatment of some neuropsychiatric disorders with the antipsychotic drugs and antidepressants targeting 5-HT2A receptors.

Serotonin 2C Receptor Alternative Splicing in a Rat Model of Orofacial Neuropathic Pain

Abnormal serotonin 2C receptor (5HTR2C) alternative splicing and RNA editing are involved in the etiology of pain disorders. Functional 5HTR2C can only be generated when alternative exon Vb is included within the mRNA;the small nucleolar RNA RBII-52 is complementary to exon Vb and promotes its inclusion. The expression of HBII-52 (the human equivalent of RBII-52) is reduced in Prader-Willi syndrome, patients of which have a high pain threshold. Here, we measured the pain threshold in a rat model of orofacial neuropathic pain and related it to the expression levels of wild-type and variant 5HTR2C and RBII-52. We generated an infraorbital nerve loose ligation model of neuropathic pain in rats and measured the pain threshold of the animals using mechanical stimulation with von Frey filaments. We then sacrificed the animals and examined the RNA levels of 5HTR2C and RBII-52 in the cervical spinal cord by real-time PCR. On post-injury day 28, pain threshold values in injured rats were significantly lower than in sham-operated or na?ve animals. The levels of total and exon Vb-skipped 5HTR2C mRNA were significantly lower in injured rats than in that sham-operated or na?ve rats, and the ratio of exon Vb-skipped 5HTR2C to total 5HTR2C was significantly higher. There were no significant differences in RBII-52 expression among the groups. Our data suggest that neuropathic pain induces serotonergic dysfunction mediated by 5HTR2C alternative splicing. 5HTR2C might be subject to complicated and fine regulation both by RNA editing and by alternative splicing.

5-羟色胺2C受体对形觉剥夺性成年弱视大鼠离体脑片初级视皮层长时程增强的影响

目的探讨5-羟色胺2C受体(5-HT2CR)对形觉剥夺性成年弱视大鼠离体脑片视皮层V1M区长时程增强(LTP)的影响。方法将16只2周龄Spargue Dawley大鼠随机分为正常对照组和单眼形觉剥夺组,每组8只。正常对照组大鼠不做任何处理;单眼形觉剥夺组大鼠行右侧眼睑缝合术制备单眼形觉剥夺性弱视模型,造模成功后饲养6周,然后处死2组大鼠,冠状切取400μm厚视皮层脑组织切片并孵育于人工脑脊液中。根据人工脑脊液中加入的药物不同,将正常对照组大鼠视皮层切片作为A组,将剥夺眼对侧视皮层切片分为B、C、D和E组,将剥夺眼同侧视皮层切片分为F、G、H和I组。A、B、F组人工脑脊液中不加任何药物,C、G组加生理盐水,D、H组加10μmol·L-15-羟色胺盐酸盐,E、I组加10μmol·L-1SB 242084和10μmol·L-15-羟色胺盐酸盐。采用细胞外微电极记录法对各组大鼠视皮层组织切片进行电生理学实验,记录离体组织切片视皮层V1M区LTP并计算神经元场突触后电位(f PSP)斜率。结果 A、B、C、D、E、F、G、H、I组大鼠视皮层f PSP斜率分别为(198.1±13.5)%、(106.3±8.3)%、(106.3±8.3)%、(157.1±9.7)%、(102.6±4.7)%、(144.5±2.9)%、(144.5±2.9)%、(192.2±8.6)%和(129.7±13.5)%。A、B、F组大鼠视皮层f PSP斜率两两比较差异均有统计学意义(P<0.001),其中A组大鼠视皮层f PSP斜率高于B、F组(P<0.001),B组大鼠视皮层f PSP斜率低于F组(P<0.001)。D组大鼠视皮层f PSP斜率高于C组(t=-10.833,P<0.001);H组大鼠视皮层f PSP斜率高于D、G组(t=-6.841、-10.616,P<0.001);E组大鼠视皮层f PSP斜率低于D、I组(t=11.872、-3.910,P<0.001,P<0.05);I组大鼠视皮层f PSP斜率低于H组(t=9.911,P<0.001)。结论单眼形觉剥夺可造成双侧视皮层神经元功能减退,5-羟色胺盐酸盐可通过5-HT2CR起到一定逆转作用。

5-羟色胺2C受体作为精神分裂症治疗靶点的研究进展

随着第2代抗精神病药物引起体质量增加和心血管不良反应的报道越来越多,研究者近年来开始将关注点投向治疗精神分裂的新靶点——5-羟色胺2C受体(5-HT2C受体)。一方面,有研究表明激动5-HT2C受体具有降低黑质纹状体多巴胺的作用,推测其潜在的抗精神病作用;另一方面,第2代抗精神病药物对5-HT2C受体的拮抗作用是其引起患者体质量增加的重要因素,反之激动5-HT2C受体则可能具有降低体质量的效应。该文以文献综述的形式就5-HT2C受体的抗精神病效应和与之相关的体质量增加不良反应进行阐述。

Synergistic interaction between C5a and NOD2 signaling in the regulation of chemokine expression in RAW 264.7 macrophages

The innate immune response is a complex process involving multiple pathogen-recognition receptors, including toll-like receptors (TLRs) and nucleotide-binding oligomerization domain (NOD)-like receptors. Complement is also a critical component of innate immunity. While complement is known to interact with TLR-mediated signals, the interactions between NOD-like receptors and complement are not well understood. Here we report a synergistic interaction between C5a and Nod2 signaling in RAW 264.7 marophages. Long-term treatment with muramyl dipeptide (MDP), a NOD2 ligand, enhanced C5a-mediated expression of chemokine mRNAs in RAW 264.7 cells. This response was dependent on NOD2 expression and was associated with a decrease in expression of C5L2, a receptor for C5a which acts as a negative modulator of C5a receptor (C5aR) activity. MDP amplified C5a-mediated phosphorylation of p38 MAPK. Treatment of RAW264.7 cells with an inhibitor of p38 attenuated the synergistic effects of C5aon MDP-primed cells on MIP-2, but not MCP-1, mRNA. In contrast, inhibition of AKT prevented C5a stimulation of MCP-1, but not MIP-2, mRNA, in MDP-primed cells. Taken together, these data demonstrated a synergistic interaction between C5a and NOD2 in the regulation of chemokine expression in macrophages, associated with a down-regulation of C5L2, a negative regulator of C5a receptor activity.

乳腺癌患者血清NR3C2和DNMT3A表达水平及其诊断价值研究

目的 探讨乳腺癌患者血清核受体亚家族C组成员2(nuclear receptor subfamily 3, group C, member 2,NR3C2),DNA甲基化转移酶-3A[DNA(cytosine-5)-methyltransferase 3A,DNMT3A]水平及其临床诊断价值。方法收集复旦大学附属华东医院2017年5月~2020年12月期间住院的94例乳腺癌患者为乳腺癌组,另选取同期健康体检者86例作为对照组。酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测两组血清NR3C2和DNMT3A水平,Pearson法分析乳腺癌患者血清NR3C2和DNMT3A表达水平相关性,多因素Logistic回归分析乳腺癌发生的影响因素,受试者工作特征(receiver operating characteristic,ROC)曲线评估NR3C2和DNMT3A对乳腺癌的诊断价值。结果 乳腺癌组患者血清NR3C2水平为317.84±33.47 ng/L,低于对照组(374.25±47.72ng/L),DNMT3A的表达水平为451.63±75.47μg/L,高于对照组(349.85±63.72μg/L),差异有统计学意义(t=9.243,9.729,均P<0.05)。Pearson分析结果显示,乳腺癌患者血清NR3C2与DNMT3A表达水平存在明显负相关(r=-0.501,P=0.000)。Logistic回归分析结果显示,NR3C2高水平为乳腺癌发生的保护因素(OR=0.563,95%CI:0.372~0.851,P=0.006),DNMT3A高水平是乳腺癌发生的危险因素(OR=1.834,95%CI:1.249~2.693,P=0.002)。ROC曲线结果发现,血清NR3C2与DNMT3A诊断乳腺癌的曲线下面积(area under the curve,AUC)分别为0.853(0.791~0.915),0.930(0.896~0.965),当两者联合时其AUC为0.969(0.949~0.990),高于两者单独检测(Z=3.460,1.894,P=0.000,0.034)。结论 乳腺癌患者血清NR3C2表达水平降低,DNMT3A表达水平升高,两者联合检测能够提高乳腺癌的诊断价值。

Changes of Serotonin （5-HT）, 5-HT2A Receptor, and 5-HT Transporter in the Sprague-Dawley Rats of Depression, Myocardial Infarction and Myocardial Infarction Co-exist with Depression

Background： To evaluate whether serotonin （5-HT）, 5-HT2A receptor （5-HT2AR）, and 5-HT transporter （serotonin transporter [SERT]） are associated with different disease states of depression, myocardial infarction （MI） and MI co-exist with depression in Sprague-Dawley rats. Methods： After established the animal model of four groups include control, depression, MI and MI with depression, we measured 5-HT, 5-HT2AR and SERT from serum and platelet lysate.Results： The serum concentration of 5-HT in depression rats decreased significantly compared with the control group （303.25 ± 9.99 vs. 352.98 ±13.73; P =0.000）, while that in MI group increased （381.78 ±14.17 vs. 352.98 ±13.73; P = 0.000）. However, the depression ＋ MI group had no change compared with control group （360.62 ±11.40 vs. 352.98 ±13.73; P = 0.036）. The changes of the platelet concentration of 5-HT in the depression, MI, and depression ＋ MI group were different from that of serum. The levels of 5-HT in above three groups were lower than that in the control group （380.40 ± 17.90, 387.75 ±22.28,246.40 ±18.99 vs. 500.29 ±20.91 ; P = 0.000）. The platelet lysate concentration of 5-HT2AR increased in depression group, MI group, and depression ＋ MI group compared with the control group （370.75 ±14.75,393.47 ±15.73,446.66 ±18.86 vs. 273.66 ±16.90; P= 0.000）. The serum and platelet concentration of SERT in the depression group, MI group and depression ＋ MI group were all increased compared with the control group （527.51 ±28.32, 602.02 ±23.32, 734.76 ±29.59 vs. 490.56 ±16.90; P 0,047, P = 0.000, P = 0.000 in each and 906.38 ±51.84, 897.33 ±60.34, 1030.17 ±58.73 vs. 708.62 ±51.15; P = 0.000 in each）. Conclusions： The concentration of 5-HT2AR in platelet lysate and SERT in serum and platelet may be involved in the pathway of MI with depression. Further studies should examine whether elevated 5-HT2AR and SERT may contribute to the biomarker in MI patients with depression.

Bioassay-guided isolation of saikosaponins with agonistic activity on 5-hydroxytryptamine 2C receptor from Bupleurum chinense and their potential use for the treatment of obesity

5-Hydroxytryptamine 2C(5-HT2C) receptor is one of the major targets of anti-obesity agents, due to its role in regulation of appetite. In the present study, the 70% EtO H extract of the roots of Bupleurum chinense was revealed to have agonistic activity on 5-HT2 C receptor, and the subsequent bioassay-guided isolation led to identification of several saikosaponins as the active constituents with 5-HT2 C receptor agonistic activity in vitro and anti-obesity activity in vivo. The new compound, 22-oxosaikosaponin d(1), was determined by extensive spectroscopic analyses(HR-ESI-MS, IR, and 1D and 2D NMR). The primary structure-activity relationship study suggested that the intramolecular ether bond between C-13 and C-28 and the number of sugars at C-3 position were closely related to the 5-HT2 C receptor agonistic activity. Saikosaponin a(3), the main saponin in B. chinense, showed obviously agonistic activity on 5-HT2 C receptor with an EC50 value of 21.08 ± 0.33 μmol×L^(–1) in vitro and could reduce food intake by 39.1% and 69.2%, and weight gain by 13.6% and 16.4%, respectively, at 3.0 and 6.0 mg×kg^(–1) in vivo. This investigation provided valuable information for the potential use of B. chinense as anti-obesity agent.

Follicular helper T lymphocytes in health and disease

A correct antibody response requires the participation of both B and T lymphocytes and antigen presenting cells. In this review we address the role of follicular helper T lymphocytes(T FH) in this reaction. We shall focus on the regulation of their development and function in health and disease. T FH can be characterized on the basis of their phenotype and the pattern of secretion of cytokines. This fact is useful to study their participation in the generation of antibody deficiency in primary immunodeficiency diseases such as common variable immunodeficiency, X-linked hyper Ig M syndrome orX-linked lymphoproliferative disease. Increased numbers of T FH have been demonstrated in several autoimmune diseases and are thought to play a role in the development of autoantibodies. In chronic viral infections caused by the human immunodeficiency virus, hepatitis B or C virus, increased circulating T FH have been observed, but their role in the protective immune response to these agents is under discussion. Likewise, an important role of T FH in the control of some experimental protozoan infections has been proposed, and it will be important to assess their relevance in order to design effective vaccination strategies.

食管癌患者肿瘤组织CXCL5和CXCR2的表达与病情和预后的相关性分析

目的研究食管癌患者肿瘤组织CXC趋化因子5(CXCL5)和CXC趋化因子受体2(CXCR2)水平对食管癌病情及预后的影响,为食管癌临床诊断、治疗和评价预后提供新方向。方法选取2013年8月至2015年7月于徐州医科大学附属医院确诊且临床资料完整的126例未经放化疗的食管癌患者作为研究对象,通过免疫组织化学染色的方法检测食管癌组织CXCL5和CXCR2表达水平,探讨CXCL5和CXCR2表达水平与食管癌临床病理分型和生存预后的关系。结果食管癌组织CXCL5和CXCR2的表达阳性率显著高于癌旁组织(P<0.05),且两者存在相关性(P<0.05);食管癌组织CXCL5和CXCR2与食管癌TNM分期和淋巴结转移具有显著的相关性(P<0.05);生存曲线显示,食管癌组织CXCL5、CXCR2阳性的患者5年生存率和预期生存时间显著低于阴性者,差异有统计学意义(P<0.05);Cox多因素分析显示原发肿瘤部位、食管癌TNM分期、淋巴结转移和食管癌组织CXCL5、CXCR2水平为食管癌预后的独立危险因素(P<0.05)。结论食管癌患者肿瘤组织内CXCL5、CXCR2高表达,与食管癌病情相关,CXCL5、CXCR2是影响食管癌患者预后的独立危险因素。

Extract of Yokukansan improves anxiety-like behavior and increases serum brain-derived neurotrophic factor in rats with cerebral ischemia combined with amyloid-β42 peptide

OBJECTIVE: To examine the effects of Yokukansan(YKS) extract on two endogenous modulators of anxiety, brain-derived neurotrophic factor(BDNF)and serotonin (5-HT)_(2A) receptors pharmacologically, in the ischemic rat model of dementia.METHODS: The cerebral ischemia(CI) was induced by bilateral occlusion of the vertebral and common carotid arteries(4-vessel occlusion ischemia). The CI was combined with the amyloid-β42 peptide(A42) injected intracerebroventricularly, and referreβdto as CI+Aβ. Anxiety-like behaviors were assessed by elevated plus maze(enclosed arm), light/dark transition test(dark chamber), and open-field test.Wet-dog shakes were induced by the 5-HT_(2A) receptor agonist 2, 5-dimethoxy-4-iodoamphetamine(DOI). The concentration of BDNF in serum was determined by enzyme-linked immuno sorbent assay.RESULTS: CI + Aβ increased anxiety, as demonstrated by the increase of time spent in the enclosed arms and dark chambers, and locomotion in the outer zone of the open field(thigmotaxis). CI + Adecreased the serum concentration of BDNF. YKS reβ-duced the anxiety-like behaviors, suppressed the DOI-induced wet-dog shakes and increased serum BDNF concentrations.CONCLUSION: Our findings suggest that YKS extract improves CI + Aβ-induced anxiety by antagonizing 5-HT_(2A) receptors and increasing BDNF.

Antibody drug conjugate development in gastrointestinal cancers:hopes and hurdles from clinical trials

Gastrointestinal(GI)cancers represent the leading cause of cancer-related mortality worldwide.Antibody drug conjugates(ADCs)are a rapidly growing new class of anti-cancer agents which may improve GI cancer patient survival.ADCs combine tumour-antigen specific antibodies with cytotoxic drugs to deliver tumour cell specific chemotherapy.Currently,only two ADCs[brentuximab vedotin and trastuzumab emtansine(T-DM1)]have been Food and Drug Administration approved for the treatment of lymphoma and metastatic breast cancer,respectively.Clinical research evaluating ADCs in GI cancers has shown limited success.In this review,we will retrace the relevant clinical trials investigating ADCs in GI cancers,especially ADCs targeting human epidermal growth receptor 2,mesothelin,guanylyl cyclase C,carcinogenic antigen-related cell adhesion molecule 5(also known as CEACAM5)and other GI malignancy specific targets.We will review potential hurdles for their success and provide new perspective for future treatment.

电针足三里对严重烫伤致大鼠急性肺损伤的影响

目的 探讨电针足三里对严重烫伤致大鼠急性肺损伤的影响.方法 雄性SD大鼠40只,体重200～250 g,随机分为5组（n=8）：对照组、烫伤组、足三里组、非经非穴组和α-银环蛇毒素组（α-BGT组）.对照组尾静脉注射生理盐水1 ml.烫伤组、足三里组、非经非穴组和α-BGT组先制备30%总体表面积Ⅲ度烫伤模型,然后烫伤组尾静脉注射生理盐水1 ml;足三里组于双侧足三里穴垂直进针7 mm,给予脉冲电流（电压3V,电流2ms,频率3 Hz）持续刺激12 mim,间隔8 h刺激1次,持续2 d;非经非穴组于双侧足三里穴旁5mm处给予脉冲刺激,方法同足三里组;α-BGT组尾静脉注射α-BGT 1.0 μg/kg,再于双侧足三里穴给予脉冲刺激,方法同足三里组.各组处理结束后,处死大鼠,取肺组织,光镜下观察病理学结果,电镜下观察超微结构,采用ELISA法测定肺组织高迁移率族蛋白B1（HMGBl）含量,采用免疫组化法测定HMGBl蛋白表达,采用RT-PCR法测定HMGBl mRNA表达.结果 烫伤组肺组织光镜下可见肺泡壁崩解,泡内大量渗出液,间质水肿、肥厚和增生,伴大量炎性细胞浸润;电镜下可见细胞核形态不规则,核膜僵硬,部分凸凹不平和核溶解,胞质内板层小体明显减少,肺组织病理损伤程度较对照组减轻.与对照组比较,烫伤组、非经非穴组和α-BGT组肺组织HMGBl含量升高,HMGBl蛋白及其mRNA的表达上调（P〈0.05）,足三里组各指标差异无统计学意义（P〉0.05）;与烫伤组比较,足三里组肺组织HMGBl含量降低,HMGBl蛋白及其mRNA的表达下调,非经非穴组和α-BGT组肺组织HMGBl mRNA表达下调（P〈0.05）;与足三里组比较,非经非穴组和α-BGT组肺组织HMGBl含量升高,HMGBl蛋白及其mRNA的表达上调（P〈0.05）.结论 电针足三里可减轻严重烫伤致大鼠急性肺损伤,其机制与激活含α7亚基N型胆碱能受体介导的胆碱能抗炎通路,抑制肺组织HMGBl的表达有关.