Effect of α-synuclein on the promoter activity of tyrosine hydroxylase gene

Objective To approach the associated mechanism by which α-synuclein （α-Syn） might regulate the metabolism of dopamine. Methods A DNA fragment, located at --495 to ＋25 of the human tyrosine hydroxylase （TH） gene, was amplified by PCR and inserted into the pGL3-Basic luciferase reporter vector. The recombinant plasmid pGL3-THprom was transfected into a dopammergic cell line MES23.5 or a α-Syn over-expressed MES23.5 （named MES23.5/hα-Syn^＋）. The promoter activity was detected by the Dual Luciferase Assay System. Results The luciferase activities in the MES23.5 cells transfected with pGl.,3-Basic, pGL3-THprom, and pGL3-Control vectors were 5.60±0.67, 26.80±4.11, and 32.90±4.75, respectively. On the other hand, the luciferase activity of pGL3-THprom in the MES23.5 （26.80±4.11） was significantly higher than that in the MES23.5/hα-Syn^＋（14.40±0.61） （P〈0.01）. Conclusion These results indicate that the -495 to ＋25 region in the TH gene possesses promoter activity for controlling the gene expression, and that α-Syn may negatively regulate the metabolism of dopamine by affecting the function of TH promoter as a trans-acting factor.

Verbascoside promotes the regeneration of tyrosine hydroxylase-immunoreactive neurons in the substantia nigra

Tyrosine hydroxylase is a key enzyme in dopamine biosynthesis. Change in tyrosine hydroxylase expression in the nigrostriatal system is closely related to the occurrence and development of Parkinson＇s disease. Verbascoside, an extract from Radix Rehmanniae Praeparata has been shown to be clinically effective in treating Parkinson＇s disease. However, the underlying mechanisms remain unclear. It is hypothesized that the effects of verbascoside on Parkinson＇s disease are related to tyrosine hydroxylase expression change in the nigrostriatal system. Rat models of Parkinson＇s disease were established and verbascoside（60 mg/kg） was administered intraperitoneally once a day. After 6 weeks of verbascoside treatment, rat rotational behavior was alleviated; tyrosine hydroxylase m RNA and protein expression and the number of tyrosine hydroxylase-immunoreactive neurons in the rat right substantia nigra were significantly higher than the Parkinson＇s model group. These findings suggest that the mechanism by which verbascoside treats Parkinson＇s disease is related to the regeneration of tyrosine hydroxylase-immunoreactive neurons in the substantia nigra.

Dark rearing maintains tyrosine hydroxylase expression in retinal amacrine cells following optic nerve transection

The present study examined changes in retinal tyrosine hydroxylase （TH） expression in rats having undergone optic nerve transection and housed under a normal day/night cycle or in the dark. The aim was to investigate the effects of amacrine cells on axonal regeneration in retinal ganglion cells and on the synapses that transmit visual signals. The results revealed that retinal TH expression gradually decreased following optic nerve transection in rats housed under a normal day/night cycle reaching a minimum at 5 days. In contrast, retinal TH expression decreased to a minimum at 1 day following optic nerve transection in dark reared rats, gradually increasing afterward and reaching a normal level at 5 7 days. The number of TH-positive synaptic particles correlated with the TH levels indicating that dark rearing can help maintain TH expression during the synaptic degeneration stage （5 7 days after optic nerve injury） in retinal amacrine cells.

Isolation and Expression Analysis of Two Genes Encoding Cinnamate 4-Hydroxylase from Cotton(Gossypium hirsutum)

Two genes （GhC4H1 and GhC4H2） that encode putative cotton cinnamate 4-hydroxylases that catalyze the second step in the phenylpropanoid pathway were isolated from developing cotton fibers. GhC4H1 and GhC4H2 each contain open reading frames of 1 518 base pairs （bp） in length and both encode proteins consisting of 505 amino acid residues. They are 90.89% identical to each other at the amino acid sequence level and belong to class I of plant C4Hs. GhC4H1 and GhC4H2 genomic DNA are 2 247 and 2 161 bp long, respectively, and contain two introns located at conserved positions relative to the coding sequence. GhC4HI and GhC4H2 promoters were isolated and found to contain many cis-elements （boxes P, L and AC-1 element） previously identified in the promoters of other phenylpropanoid pathway genes. Histochemical staining showed GUS expression driven by the GhC4H1 and GhC4H2 promoters in ovules and fibers tissues. GhC4H1 and GhC4H2 were also widely expressed in other cotton tissues. GhC4H2 expression reached its highest level during the elongation stage of fiber development, whereas GhC4H1 expression increased during the secondary wall development period in cotton fibers. Our results contribute to a better understanding of the biochemical role of GhC4H1 and GhC4H2 in cotton fiber development.

Tyrosine Hydroxylase as a Target for Deltamethrin in the Nigrostriatal Dopaminergic Pathway

Objective To study the effects of deltamethrin on tyrosine hydroxylase in nigrostriatum of male rats. Methods Sprague-Dawley rats were daily treated with deltamethrin at 6.25 or 12.5 mg/kg body weight by gavage for 10 days. Then HPLC-fluorescence detection was used to analyze the contents of dopamine （DA）, 3,4-dihydmxyphenylacetic acid （DOPAC） and homoranillic acid （HVA） in substantial nigra and striatum. The activities of tyrosine hydroxylase （TH） were also detected by HPLC-fluorescence detection. TH mRNA or TH protein levels were measured by RT-PCR and immunohistochemistry method. Results The content of DA in stfiatum was significantly decreased by the treatments, suggesting an inhibition of DA synthesis by deltamethrin. The contents of DA metabolites DOPAC and HVA increased, indicating increased dopamine turnover. Furthermore, deltamethrin significantly decreased the activity, as well as the mRNA and protein levels of TH. Conclusions These findings reveal a novel aspect of deltamethfin neurotoxicity and suggest tyrosine hydroxylase as a molecular target of deltamethin on dopamine metabolism in the nigrostriatal pathway.

Tyrosine hydroxylase expression in the midbrain of Parkinson's disease model rats treated with Xifeng Dingchan decoction

This study showed that abnormal behavioral changes were greatly improved in rats displaying Parkinson＇s disease-like symptoms after intragastric administration of Xifeng Dingchan decoction at 15, 7.5, 3.75 g/kg per day. In addition, tyrosine hydroxylase mRNA expression in the substantia nigra of the midbrain was up-regulated, and tyrosine hydroxylase content in the midbrain ventral tegmentum and substantia nigra pars compacta was also increased. The effect of administration of Xifeng Dingchan decoction at 7.5 g/kg per day was similar to that of Madopar at 67.5 mg/kg per day. These results indicate that the therapeutic effect of Xifeng Dingchan decoction on Parkinson＇s disease is associated with the up-regulated protein and mRNA expression of tyrosine hydroxylase in the midbrain.

Treatment time influences the effects of a low-frequency pulsed electric field on synthesis of tyrosine hydroxylase and dopamine in PC12 cells

BACKGROUND： Electromagnetic radiation can influence dopamine （DA） synthesis in brain tissues or ceils, but electromagnetic frequencies, intensities, and radiation time can produce different effects. In addition, the signal pathway by which electromagnetic radiation influences DA synthesis remains controversial. OBJECTIVE： To determine tyrosine hydroxylase （TH） expression in PC12 cells and DA levels in cell culture media after different periods of low-frequency pulsed electric field （LF-PEF） stimulation, and to determine how LF-PEF signaling stimulates TH synthesis using inhibitors. DESIGN, TIME AND SETTING： A parallel, controlled, cell experiment was performed at the Laboratory of Cell Biology, School of Life Science, East China Normal University, between January and October 2006. MATERIALS： PC12 cells were purchased from the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, China. Nerve growth factor was purchased from PeproTech, USA. The protein kinase A inhibitor, H-89, and mitogen-activated protein kinase kinase inhibitor, U0126, were purchased from Sigma, USA. METHODS： （1） Following routine culture in Dulbecco＇s modified eagle medium, primary PC12 cells were stimulated under LF-PEF （pulse frequency 50.Hz, pulse width 20 μs, peak field strength 1 V/m） for 5, 10, 15, 20, and 30 minutes. （2） Inhibitors （H-89 or U0126, 1 μmol/L） were added 30 minutes before LF-PEF stimulation for 10 minutes. MAIN OUTCOME MEASURES： （1） TH expression was determined by Western blot in PC12 cells at 0.5, 1,2, 3, and 4 days after LF-PEF stimulation. Similarly, DA was measured by high-performance liquid chromatography in media at 2, 3, 4, or 5 days after LF-PEE （2） TH expression was detected 1 day after H-89 or U0126 treatment and LF-PEE RESULTS： （1） Short-term LF-PEF stimulation （5 and 10 minutes） increased TH expression and media DA levels after short-term culture （2 days） （P 〈 0.01）, but both parameters decreased with longer culture （3 4 days） （P 〈 0.01）. Long-term LF-PEF stimulation （15, 20, or 30 minutes） decreased TH and DA synthesis, followed by a rapid increase （P 〈 0.01）. （2） H89 could completely inhibit TH expression in PC12 cells stimulated by LF-PEF for 10 minutes, while the inhibition rate of U0126 was 53.2%. CONCLUSION： Short-term LF-PEF first promotes then inhibits, while long-term LF-PEF first inhibits then promotes, TH and DA synthesis. LF-PEF stimulation regulates TH expression primarily by activating protein kinase A to regulate DA synthesis.

Tyrosine hydroxylase and Lewy body molecules immunoreactivity in the SNC neurons of an AS/AGU mutantrat

The AS/AGU rat has a recessive single point mutation in the gene coding for the gamma isoform of protein kinase C (PKC-γ) resulting in a failure to release dopamine in the striatum and impaired movement including a staggering gait, difficulty in initiating movement and a slight whole body tremor. This study examined the levels tyrosine hydroxylase, ubiquitin and parkin in individual SNC cell bodies, there was no evidence of a reduction in tyrosine hydroxylase levels although levels of ubiquitin and parkin were elevated in the cytoplasm. The findings support the hypothesis that the initial bar to dopamine availability in the striatum is reduced release, with substantia nigra cell death being a later phenomenon.

COEXISTENCE OF FOS-LIKE PROTEIN IN TYROSINE HYDROXYLASE-LIKE IMMUNOREACTIVE NEURONS IN THE HINDBRAIN OF ARTS FOLLOWING PERIPHERAL ELECTRICAL STIMULATION

In the present study we have found that proto-oncogene c-fos protein can expressin the noradrenergic neurons of rat hindbrain following peripheral electrical stimulation. Ratswere given peripheral electrical stimulation via thin stainless steel pins inserted into the pointsnear knee joint (S36) and ankle joint (Sp6) which mimic the manipulation of electroacupuncture(EA) performed in humans. Animals were perfused for double staining immunohistochemistry 2hafter the termination of EA. In rats subjected to EA stimulation Fos-like protein was found in thetyrosine hydroxylase (TH)-like immunoreactive neurons in rat hindbrain. The Fos and TH coex-isting neurons were distributed in the locus coeruleus, solitary tract nucleus, ventrolateral medul-la, periaqeductal gray, as well as superior colliculus. The percentage of the coexisting neuronscompared with the total number of neurons containing Fos-like protein in these nuclei rangedfrom 6% to 32%. The results suggest that the noradrenergic neurons in these regions may be ac-tivted by acupuncture stimulation.

Tyrosine hydroxylase gene transfections to different sites of striatum in the rat model of Parkinson’s disease

The use of gene therapy has been intensively studied as a potential method to treat Parkinson’s disease (PD) and other degenerative brain diseases. However, the effects of experimental measures and approaches on the outcome of gene delivery or on the physiological state of target tissues have not been analyzed as much and systematically. Therefore, we have infused adenovirus vectors expressing either a therapeutic tyrosine hydroxylase (TH) gene or a lacZ reporter gene into striatum in a rat model of PD. The experimental procedures were tested using the Ad lacZ vector in order to optimize concentrations, volumes, infusion speeds and transfection times. The expression of Ad lacZ vector was lower and declined earlier in the lesioned than unlesioned striatum suggesting that the lesion affects on the transfection efficiency and outcome of gene transfection. The effect of three different approaches of Ad TH vector transfection was compared: 1) the delivery of Ad TH gene vector alone into one single site of striatum, 2) the delivery of Ad TH gene vector alone into multiple sites of striatum, and 3) the delivery of Ad TH gene vector into one site of striatum followed by a continuous infusion of tetrahydrobiopterin (BH4) cofactor with a mini pump. There was a small and transient unsignificant decrease in the turning behavior when the Ad TH vector was delivered into one site of the striatum. Simultaneous infusion into several sites or together with BH4 cofactor did not improve more the effect of gene delivery. Thus, although the effects were unsignificant, the Ad TH transfection seemed to decrease the turning behavior in the rat model of PD and the optimal effect was seen at some specific doses and time points. Furthermore, the outcome of gene therapy could depend in addition to the amount and efficacy of gene vectors also on the physiological state and experimental strategies.

Structural Analysis of an Oligosaccharide and Glycopeptide Mixture from Panax Ginseng Root with Inhibition SHP-1 Function

A mixture of oligosaccharide and glycopeptide was isolated from the aqueous extract of Panax ginseng roots. The mixture inhibits protein tyrosinc phosphatase（SHP-1） function, implying it enhances immune activity. The peak molecular mass of the oligosaccharide portion is 1800 calculated via GPC software after separation by HPLC. And the structure of the oligosaccharide portion is the backbone of （1→3）- and （1→4）-linked arabinopyranoside, and （1→4）- and （1→6）-linked glucopyranoside, with non-reducing terminals of arabinopyranoside and glucopyranoside. The peak molecular mass of glycopeptide portion is 1900 calculated via GPC software after separation by HPLC. The structure of glycopeptide portion is the backbone of （1→3）- and （1→4）-linked arabinopyranoside, and （1→3,6）-linked glucopyranoside, with non-reducing terminals of galactopyranose and glucopyranoside. The peptide composition is Glu. Asp, Hyp, Set, Arg, Gly , Thr, Pro, Ala, Val, lie, Leu and Lys. The oligosaccharide-peptide linkage is formed by Ara and Hyp.

表皮生长因子受体酪氨酸激酶抑制剂联合化疗一线治疗表皮生长因子受体突变晚期非小细胞肺癌疗效及安全性的Meta分析

背景表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)的靶向治疗已成为表皮生长因子受体(EGFR突变晚期非小细胞肺癌(NSCLC)的规范治疗方案,但临床发现了不可避免的原发性或继发性耐药最终导致了疾病进展。为此,寻找早期预测有效人群的标志物和探索延长或逆转继发性耐药的优化治疗方案成为国内外研究重点。目的依据国内外文献数据系统评价EGFR-TKI联合化疗一线治疗EGFR突变晚期NSCLC的疗效及安全性。方法计算机检索PubMed、Embase、Cochrane Library数据库发布的有关EGFR-TKI联合化疗对比单药EGFR-TKI一线治疗EGFR突变晚期NSCLC患者疗效及安全性的随机对照试验,检索时间为建库至2023年11月。由2名研究者独立筛选文献、提取数据并评价纳入研究的偏倚风险,对无进展生存期(PFS)、总生存期(OS)、客观缓解率(ORR)、疾病控制率(DCR)以及≥3级不良反应等数据收集和分析。基于基线临床特征进行亚组分析,使用RevMan 5.4.1版本进行数据统计分析。结果纳入符合条件的10项研究,共2029例患者,其中EGFR-TKI联合化疗组1049例患者,单纯EGFR-TKI组980例患者。Meta分析结果显示,与单纯EGFR-TKI组相比,EGFR-TKI联合化疗组可延长EGFR突变晚期NSCLC患者的PFS(HR=0.54,95%CI=0.49~0.60,P<0.00001)和OS(HR=0.69,95%CI=0.59~0.79,P<0.00001)。与单纯EGFR-TKI组相比,EGFR-TKI联合化疗组可提高EGFR突变晚期NSCLC患者的ORR(OR=1.95,95%CI=1.57~2.42,P<0.00001)和DCR(OR=1.76,95%CI=1.13~2.74,P=0.01)。在伴随脑转移的患者中,与单纯EGFR-TKI治疗相比,EGFR-TKI联合化疗延长EGFR突变晚期NSCLC患者的PFS(HR=0.42,95%CI=0.34~0.52,P<0.00001)和OS(HR=0.69,95%CI=0.51~0.94,P=0.02)。在基线无脑转移的患者中,与单纯EGFR-TKI治疗相比,EGFR-TKI联合化疗延长EGFR突变晚期NSCLC患者的PFS(HR=0.62,95%CI=0.53~0.72,P<0.00001)。EGFR-TKI联合化疗治疗EGFR突变晚期NSCLC患者的≥3级不良反应发生率高于单纯EGFR-TKI治疗(OR=4.25,95%CI=2.74~6.61,P<0.00001)。结论EGFR-TKI联合化疗可显著延长EGFR突变晚期NSCLC患者的一线PFS及OS,尤其是基线脑转移患者;但不良反应发生率增加,尤其是≥3级血液学相关毒副作用发生率,整体是安全可控的。

Unraveling the mechanism of action of Shangxia Liangji formula for treating insomnia:a metabolomics and network pharmacology approach

Background:Insomnia is a prevalent clinical condition and Shangxia Liangji formula(SXLJF)is a well-established method of treatment.Nevertheless,the specific mechanism of action of SXLJF remains unclear.Methods:The mouse model of insomnia was established by intraperitoneal injection of para-chlorophenylalanine.Forty-two mice were randomly divided into a negative control group,model group,SXLJF group(18.72 g/kg/day),and positive control group(diazepam,2 mg/kg)and treated with the corresponding drugs for 7 consecutive days.The open field test and pentobarbital-induced sleeping test were conducted.LC-MS-based untargeted metabolomics and network pharmacology were applied to explore the potential targets of SXLJF for treating insomnia.Finally,key targets were validated using RT-qPCR.Results:Behavioral tests demonstrated that SXLJF reduced the total distance,average velocity,central distance,and sleep latency,and prolonged sleep duration.Metabolomics and network pharmacology revealed potential targets,signaling pathways,metabolic pathways,and metabolites associated with the anti-insomnia effects of SXLJF.Specifically,tyrosine hydroxylase(TH)and tyrosine metabolism emerged as crucial metabolic pathways and targets,respectively.RT-qPCR results supported the role of TH in the mechanism of SXLJF in treating insomnia.Conclusion:In conclusion,TH and tyrosine metabolism may represent significant targets and pathways for SXLJF in treating insomnia.

Effect of Bushen Huoxue Decoction(补肾活血饮) on the Orphan Receptor and Tyrosine Hydroxylase in the Brain of Rats with Parkinson's Disease

Objective：To explore the effect of Bushen Huoxue Decoction（补肾活血饮,BHD） on the orphan receptor（Nurr1） and tyrosine hydroxylase（TH） in the brain of rats with Parkinson＇s disease（PD）.Methods：One hundred and twenty SD rats were divided into 100 in the model group and 20 in the normal control group,fifty-eight SD rats from the model group,established into PD model successfully by injuring their substantia nigra （SSN） with 6-hydroxydopamine,were divided equally into the model group and the test group,and they were treated with saline and BHD,respectively,for eight successive weeks.The change in the rats＇ behavior before and after treatment was observed by counting the cycles of rotation induced by apomorphine injection; the pathology of neurons,level of Nurr1 mRNA expression,and amount of TH positive cells in SSN were observed after treatment.Results：The rats＇ behavior was improved in the tested group significantly,the rotation cycle after treatment being 84.0±20.0 cycles/40 min,which was significantly lower than that in the model group（377.0±62.3 cycles/40 min,P〈0.01）.Besides,the Nurr1 mRNA expression and TH positive cell in the test group were 0.97±0.15 and 49.40±14.72,respectively,which were significantly higher than those in the model group,0.22±0.03 and 5.45±2.58,respectively（all P〈0.01）.Conclusion：BHD could treat PD by enhancing the Nurr1 mRNA expression,increasing the TH content in brain,and promoting the repairing of injured neuron in cerebral SSN.

Quanti?cation of Tyrosine Hydroxylase and Erb B4 in the Locus Coeruleus of Mood Disorder Patients Using a Multispectral Method to Prevent Interference with Immunocytochemical Signals by Neuromelanin

The locus coeruleus(LC) has been studied in major depressive disorder(MDD) and bipolar disorder(BD). A major problem of immunocytochemical studies in the human LC is interference with the staining of the immunocytochemical end-product by the omnipresent natural brown pigment neuromelanin. Here, we used a multispectral method to untangle the two colors: blue immunocytochemical staining and brown neuromelanin.We found significantly increased tyrosine hydroxylase(TH) in the LC of MDD patients—thus validating the method—but not in BD patients, and we did not find significant changes in the receptor tyrosine-protein kinase ErbB4 in the LC in MDD or BD patients. We observed clear co-localization of ErbB4, TH, and neuromelanin in the LC neurons. The different stress-related molecular changes in the LC may contribute to the different clinical symptoms in MDD and BD.

CRISPR/Cas9-mediated Tyrosine hydroxylase knockout resulting in larval lethality in Agrotis ipsilon

Tyrosine hydroxylase (TH)is involved in insect melanin and the catecholamine biosynthesis pathway.TH as an enzyme catalyzing the conversion of tyrosine to 3,4- dihydroxyphenylalanine is the first step reaction in the pathway.Although TH has been proven to affect the pigmentation of the epidermis and development in many insects,there is no report about physiological function of the THgene inAgrotis ipsilon.Here we cloned the TH gene from A.ipsilon.Semi-quantitative real-time polymerase chain reaction (PCR) analysis showed that AiTH was expressed at all development stages.Moreover,its high expression levels in the head and epidermis suggest that it is mainly related to pigment deposition and insect development.Then,we used the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9system to target the AiTH gene:deletion events were detected at the target sites.Compared with the control group,a few mutants with the phenomenon of narrowing in the egg shell and embryos can develop but cannot hatch;the other hatched embryos were seriously dehydrated after hatching and died within the first day.Quantitative real-time PCR analysis revealed that THwas down-regulated inAiTHmutants.Here,our work demonstrated thatAiTHplays an important role in growth and development of newly hatched larvae;meanwhile,it would be a promising target to explore a control strategy for A.ipsilon.

Cost-effectiveness analysis of tyrosine kinase inhibitors(erlotinib,gefitinib,afatinib and osimertinib)as first-line therapy for epidermal growth factor receptor-mutated advanced non-small cell lung cancer

Gefitinib,erlotinib,afatinib and osimertinib have been recommended as the first-line treatment for epidermal growth factor receptor(EGFR)-mutated advanced non-small cell lung cancer(NSCLC),whereas no studies have compared the cost-effectiveness of these four tyrosine kinase inhibitors(TKIs)simultaneously in China.In the present study,we aimed to estimate the cost-effectiveness of erlotinib,gefitinib,afatinib and osimertinib for untreated EGFR-mutated advanced NSCLC.A Markov model was constructed to compare the 10-year impact of four TKIs for patients with treatment-naive EGFR-mutated advanced NSCLC from the perspective of the Chinese medical system.Clinical data and utility values were derived from published literature,and costs were obtained from Chinese official websites.The primary output indicator was the incremental cost-effectiveness ratio(ICER).Sensitivity analyses were performed to test the robustness of the model.We found that afatinib was estimated to spend the lowest cost with minimum life-years(LYs),while osimertinib was the most expensive regimen with maximum LYs.The ICER of gefitinib versus afatinib was$732/quality-adjusted life-year(QALY),which was less than the willingness-to-pay(WTP)of$29382/QALY.Compared with gefitinib,erlotinib yielded a higher cost and a shorter lifetime,hence it was identified as a dominated strategy.Then,osimertinib was compared to gefitinib,which produced an ICER of$71330/QALY,exceeding the WTP.It suggested that gefitinib was the most cost-effective regimen as the first-line treatment for EGFR-mutated advanced NSCLC.Decreasing the osimertinib price or increasing the WTP threshold to$68558/QALY might enhance the favorability of the outcome,by which osimertinib might become more cost-effective.One-way sensitivity analysis manifested that the model was robust.

Effect of Acupuncture on Contents of Tyrosine Hydroxylase and Glial Fibrillary Acidic Protein in Ventral Tegmental Area of Heroin Self-administrating Rats

Objective： This present study is to investigate the biochemical adaptations in the rdated brain regions of the mesolimbic dopamine system, such as the ventral tegmental area （VFA）, nucleus accumbens （NAc）, amygdale （Amy）, prefrontal cortex （PFC） , substantia nigra （SN） and caudateputamen （CPu） in response to heroin self-administration in rats and observe the effect of electroacupuncture on them. Methods. Thirty rats were trained by nose-poking response to establish stable intravenous heroin self-administration within 14 days, and then divided randomly into model group （group B） including 6 rats, withdrawal group which were withdrawn from heroin for 1 week （group C, n = 6） and for 2 weeks （group D, n= 6）, during which time they only lived in their individual home cages, and dectrtyacupuncture group which were also withdrawn from heroin for 1 week （group E, n= 6） and for 2 weeks （group F, n = 6）, during which time they were given electro-acupuncture treatment for 20 min daily and then returned to their individual home cages; in the meantime, another 6 rats were trained by nose-poking response with saline for 14 days as control （group A）; Then the leeds of tyrosine hydroxylase （TH） and glial fibrillary acidic protein （GFAP） in VFA, NAc, Amy, PFC, SN, CPu were detected with immunohistochemistry method. Results. The leeds of TH and GFAP in VFA of the heroin self administrating rats were obviously increased, and the leeds of TH and GFAP in NAc were also decreased, and these changes were not found in SN, CPu, Amy and PFC; Electro-acupuncture could promote the up regulation of TH and GFAP in VTA and down-regulation of TH and GFAP in NAc to return to the normal leeel. Conclusions： The chronic heroin self administration produced some biochemical adaptations in the related brain regions of the mesolimbic dopamine system and electroacupuncture could promote the repair of the ＂injured＂ DA neurons in VTA of heroin addicted rats and their functional recovery.

Comparison of tyrosine hydroxylase immunoreactivity in cochleas of chinchilla and guinea pig

Objective To investigate the distribution of the tyrosine hydroxylase (TH) immunoreactivity in the cochleas of chinchilla and compare it with guinea pig. Methods Ten healthy chinchillas and six healthy guinea pigs were used in this study. The surface preparation of cochleas were obtained by microdissection. The TH like immunoreactivity on the surface preparation as examined by the avidin and biotinylated horseradish peroxidase macromolecular complex (ABC) technique and investigated with light microscopy. Results TH like immunoreactivity was seen in the region from modiolus to the habenula perforata. There are two different groups of fibers in this aera, one is blood vessel dependent, the other is blood vessel independent system. No TH like immunoreactivity was present in the inner spiral bundle (ISB) and the tunnel spiral bundle (TSB) regions in chinchilla cochleas. TH like immunoreactivity was found in the region from modiolus to the habenula perforata and the ISB and TSB regions in the organ of corti (OC) in guinea pig cochleas. Conclusions The presence of TH immunoreactivity on the cochlea of chinchilla and guinea pig is different. TH like immunoreactivity on chinchilla cochlea was involved only in the fibers corresponding to the sympathetic fibers on cochlea of guinea pig. No fibers corresponding to the dopaminergic olivocochlear lateral efferent system of guinea pig were found in ISB and TSB within the OC in chinchilla.

A core-satellite-like nanoassembly reverses a decisive tyrosine hydroxylase loss in degenerative dopaminergic neurons

In recent years,neurodegenerative diseases,such as Parkinson’s or Alzheimer’s diseases,are rapidly rising in prevalence.The main hallmark of Parkinson’s disease is the falling levels of neurotransmitter dopamine in the mid-brain with dopaminergic neurons losing.Typical therapeutic solutions,including drugs,deep brain stimulation,and cell transplantation,can only alleviate the symptoms of Parkinson’s disease.It is a tremendous challenge to reverse the function degeneration of the crucial dopaminergic neurons.Herein,we develop a core-satellite-like nanoassembly(PDA-AFn(by integrating polydopamine nanoparticles and apoferritin))to raise the expression of tyrosine hydroxylase(TH),a rate-limiting enzyme in the formation of the dopamine.Both components in the nanoassembly could cooperate with each other,not only elaborately regulate the iron homeostasis and redox microenvironment,but also utilize excessive reactive oxygen species(ROS)and iron ions in the damaged neurons to supply extra dopamine and enhance TH activity,and consequently restore the function of the degenerated neurons.Remarkably,the nanoassembly-treatment relieves the dyskinesia and dramatical increases the tyrosine hydroxylase and dopamine level in the midbrain of Parkinson’s disease model mice.It is an explicit yet inspiring advance in treatment of the neurodegeneration.