Selenoprotein P mRNA expression in human hepatic tissues

AIM: To investigate the expression of selenoprotein P mRNA (SePmRNA) in tissues of normal liver, liver cirrhosis and hepatocellular carcinoma (HCC), and its relationship with HCC occurrence and development. METHODS: The expression of SePmRNA in tissues of normal liver, liver cirrhosis and HCC were detected by in situ hybridization using a cDNA probe. RESULTS: The enzyme digesting products of PBluescript-H uman Selenoprotein P were evaluated by electrophoresis. The positive expression of SePmRNA was found in the tissues of normal liver, liver cirrhosis and HCC. The expression of SeP mRNA was found in hepatic interstitial substance, especially in endothelial cells and lymphocytes of vasculature. The positive rate of SePmRNA in normal liver tissue was 84.6% (11/13) and the positive signals appeared in the nucleus and cytoplasm, mostly in the nucleolus, and the staining granules were larger in the nucleolus and around the nucleus. The positive rate of SePmRNA in liver cirrhosis tissue was 45.0% (9/20) and the positive signals were mainly in the nucleolus and cytoplasm, being less around the nucleus and inner nucleus than that in normal liver tissue. The positive rate of SePmRNA in HCC tissue was 30.0% (9/30) and the positive signals were in the cytoplasm, but less in the nucleus. CONCLUSION: SePmRNA expression in the tissues of normal liver and HCC is significantly different (84.6% vs 30.0%, P = 0.003), suggesting that SeP might play a role in the occurrence and development of HCC.

Dual Convergence of Facial Nerve Branches Innervating Whisker Pad in Rats

The precise anatomy of the facial nerve branches innervating rat whisker pad and the distribution of their corresponding motor neurons in facial nucleus area were investigated.The extratemporal facial nerves of 6 rats were anatomically observed under a surgical microscope,and then the nerve specimens of facial nerve branches at 7 anatomical sites were taken and examined for the axons and myelin sheath using Luxol fast blue staining.The distribution of facial motor neurons innervating the facial branches was observed in 12 rats by retrograde labelling.The distal pes,a fusing architecture of the buccal and marginal mandibular branches,was found to furcate into superior,middle and inferior branches to innervate whisker pad.Histologically,the myelin sheath of each branch was morphologically consistent,and the nerve fiber bundles of facial nerve branches became increasingly thinner and scattered,particularly after crossing the distal pes site and innervating the whisker pad.The facial motor neurons innervating the buccal and marginal mandibular branches were clearly distributed in similar regions in facial nucleus.This study confirmed the highly spatial synergy between the buccal and marginal mandibular branches innervating the whisker pad from extratemporal anatomy and distribution of facial motor neurons.

Effect of brain reflex instrument combined with acupuncture on convalescent neurotrophic status and nerve cell apoptosis in patients with cerebral infarction

Objective:To study the effect of brain reflex instrument combined with acupuncture on convalescent neurotrophic status and nerve cell apoptosis in patients with cerebral infarction. Methods:A total of 116 patients with convalescent cerebral infarction who were treated in Chinese Medicine Hospital Affiliated to Xinjiang Medical University between October 2014 and December 2016 were selected and randomly divided into two groups, intervention group received brain reflex instrument combined with acupuncture intervention + conventional intervention, and the control group only received conventional intervention. Serum levels of neurotrophic cytokines, monoamine neurotransmitter, and nerve cell apoptosis molecules were detected before intervention as well as 10, 20 and 30 d after intervention.Results:Serum BDNF, VEGF, bFGF, NE, E, 5-HIAA, DOPAC, HVA and Bcl-2 levels of both groups 10, 20 and 30 d after intervention were significantly higher than those before intervention while sFas, sFasL and sTRAIL levels were significantly lower than those before intervention, and serum BDNF, VEGF, bFGF, NE, E, 5-HIAA, DOPAC, HVA and Bcl-2 levels of intervention group 10, 20 and 30 d after intervention were significantly higher than those of control group while sFas, sFasL and sTRAIL levels were significantly lower than those of control group.Conclusions: Brain reflex instrument combined with acupuncture can significantly improve the convalescent neurotrophic status and nerve cell apoptosis in patients with cerebral infarction.

Effects of a non-selective TRPC channel blocker, SKF-96365, on melittin-induced spontaneous persistent nociception and inflammatory pain hypersensitivity

Objective Melittin is the main peptide in bee venom and causes both persistent spontaneous nociception and pain hypersensitivity. Our recent studies indicated that both transient receptor potential （TRP） vanilloid receptor 1 （TRPV 1） and canonical TRPs （TRPCs） are involved in mediating the melittin-induced activation of different subpopulations of pri- mary nociceptive cells. Here, we further determined whether TRPC channels are involved in melittin-induced inflamma- tory nociceptive responses in behavioral assays. Methods The anti-nociceptive and anti-hyperalgesic effects of localized peripheral administration of three doses of the non-selective TRPC antagonist, SKF-96365 （1-{[3-[3-（4-methoxyphenyl） propoxy]-4-methoxyphenyl}-lH-imidazole hydrochloride）, were evaluated in melittin tests. Pain-related behaviors were rated by counting the number of paw flinches, and measuring paw withdrawal thermal latency （s） and paw withdrawl me- chanical threshold （g）, over a l-h time-course. Results Localized peripheral SKF-96365 given before melittin prevented, and given after melittin significantly suppressed, the melittin-evoked persistent spontaneous nociception. Pre-blockade and post-suppression of activation of primary nociceptive activity resulted in decreased hypersensitivity to both thermal and mechanical stimuli applied to the primary injury site of the ipsilateral hindpaw, despite dose-effect differences between thermal and mechanical hyperalgesia. However, local administration of SKF-96365 into the contralateral hindpaw had no significant effect on any pain-associated behaviors. In addition, SKF-96365 had no effect on baseline threshold for either thermal or mechanical sensitivity under normal conditions. Conclusion Besides TRPV1, SKF-96365-sensitive TRPC channels might also be involved in the pathophysiological processing of melittin-induced inflammatory pain and hyper- sensitivity. Therapeutically, SKF-96365 is equally effective in preventing primary thermal and mechanical hyperalgesia as well as persistent spontaneous nociception. However, this drug is likely to be more effective in the relief of thermal hyper- algesia than mechanical hyperalgesia when applied 5 min after establishment of primary afferent activation.

Ethanol Increases Mechanical Pain Sensitivity in Rats via Activation of GABA_A Receptors in Medial Prefrontal Cortex

Ethanol is widely known for its ability to cause dramatic changes in emotion, social cognition, and behavior following systemic administration in humans.Human neuroimaging studies suggest that alcohol dependence and chronic pain may share common mechanisms through amygdala-medial prefrontal cortex（m PFC） interactions. However, whether acute administration of ethanol in the m PFC can modulate pain perception is unknown.Here we showed that bilateral microinjections of ethanol into the prelimbic and infralimbic areas of the m PFC lowered the bilateral mechanical pain threshold for 48 h without influencing thermal pain sensitivity in adult rats.However, bilateral microinjections of artificial cerebrospinal fluid into the m PFC or bilateral microinjections of ethanol into the dorsolateral PFC（also termed as motor cortex area 1 in Paxinos and Watson＇s atlas of The Rat Brain. Elsevier Academic Press, Amsterdam, 2005） failed to do so, suggesting regional selectivity of the effects of ethanol. Moreover, bilateral microinjections of ethanol didnot change the expression of either pro-apoptotic（caspase-3 and Bax） or anti-apoptotic（Bcl-2） proteins, suggesting that the dose was safe and validating the method used in the current study. To determine whether c-aminobutyric acid A（GABA_A） receptors are involved in mediating the ethanol effects, muscimol, a selective GABA_Areceptor agonist, or bicuculline, a selective GABA_A receptor antagonist, was administered alone or co-administered with ethanol through the same route into the bilateral m PFC. The results showed that muscimol mimicked the effects of ethanol while bicuculline completely reversed the effects of ethanol and muscimol. In conclusion, ethanol increases mechanical pain sensitivity through activation of GABA_A receptors in the m PFC of rats.

Actomyosin is Involved in the Organization of the Microtubule Preprophase Band in Arabidopsis Suspension Cultured Cells

The microtubule preprophase bands （PPBs） participate in the sequence of events to position cell plates in most plants. However, the mechanism of PPB formation remains to be clarified. In the present study, the organization of PPBs in Arabidopsis suspension cultured cells was investigated by confocal laser scanning microscopy combined with pharmacological treatments of reagents specific for the cytoskeleton elements. Double staining of F-actin and microtubules （MTs） showed that actin filaments were arranged randomly and no colocalization with cortical MTs was observed in the interphase cells. However, cortical actin filaments showed colocalization with MTs during the formation of PPBs. A broad actin band formed with the broad MT band in the initiation of PPB and narrowed down together with the MT band to form the PPB. Nevertheless, broad MT bands were formed but failed to narrow down in cells treated with the F-actin disruptor latrunculin A. In contrast, in the presence of the F-actin stabilizer phalloidin, PPB formation did not exhibit any abnormality. Therefore, the integrity, but not the dynamics, of the actin cytoskeleton is necessary for the formation of normal PPBs. Treatment with 2, 3-butanedine monoxime, a myosin inhibitor, also resulted in the formation of broad MT bands, indicating that actomyosin may be involved in the rearrangement of MTs to form the PPBs. Double staining of MTs and myosin revealed that myosin concentrated on the PPB region during PPB formation. It is suggested that the actin cytoskeleton at the PPB site may serve as a rack to transport cortical MTs by using myosin when the broad MT band narrows down to form the PPB.

Gabapentinoid Insensitivity after Repeated Administration is Associated with Down-Regulation of theα2δ-1 Subunit in Rats with Central Post-Stroke Pain Hypersensitivity

The α2δ-1 subunit of the voltage-gated Ca2＋ channel （VGCC） is a molecular target of gabapentin （GBP）, which has been used as a first-line drug for the relief of neuropathic pain. GBP exerts its anti-nociceptive effects by disrupting trafficking of the α2δ-1 subunit to the presynaptic membrane, resulting in decreased neurotrans- mitter release. We previously showed that GBP has an anti- allodynic effect in the first two weeks; but this is followed by insensitivity in the later stage after repeated adminis- tration in a rat model of central post-stroke pain （CPSP） hypersensitivity induced by intra-thalamic hemorrhage. To explore the mechanisms underlying GBP insensitivity, the cellular localization and time-course of expression of the α2δ-1 subunit in both the thalamus and spinal dorsal horn were studied in the same model. We found that the α2δ-1 subunit was mostly localized in neurons, but not astrocytes and microglia. The level of α2δ-1 protein increased in the first two weeks after injury but then decreased in the third week, when GBP insensitivity occurred. Furthermore, the c^2g-1 down-regulation was likely caused by later neuronal loss in the injured thalamus through a mechanism other than apoptosis. In summary, the present results suggest that the GBP receptor ~2^-1 is mainly expressed in thalamic neurons in which it is up-regulated in the early stage of CPSP but this is followed by dramatic down-regulation, which is likely associated with GBP insensitivity after long-term use.

Empathic Contagious Pain and Consolation in Laboratory Rodents:Species and Sex Comparisons

Dear Editor,In the past five years,we have developed a behavioral model of empathy for pain in rats[1-5].Experimentally,at least two types of behaviors associated with empathy for pain in rats can be identified,based on the evolutionary notion of the Russian doll model[6].One has been referred to as an observer's empathic consolation,which is driven by social interaction with a demonstrator in pain[3,7-9];the other is referred to as observational contagious pain(OCP or empathic transfer of pain)from a distressed object to a witnessing subject[1-3,5].

Image-Forming Visual Basis of Empathy for Pain in Mice

Dear Editor.Empathy for distress refers to the highly evolutionarilyconserved ability of humans and other social animals to feel,recognize,and understand others’painful conditions(pain,fear,and catastrophe)[1,2]and even benefit others by releasing distress through sharing,caring,and cooperation[1].In the past decade,several types of lower empathic response have been gradually identified and characterized in laboratory rodents(rats and mice),referred to as empathic contagious pain[3-5],observational fear learning[2],and contagious itch[6].