Hyperpolarization-activated cyclic nucleotide gated cation channels and substances addiction / 中华行为医学与脑科学杂志

Substances addiction is one of the important factors that deeply affect human health.At present, there is still lack of effective treatment drugs in the clinic.Exploring mechanisms of substances addiction, finding new therapeutic targets and developing effective therapeutic drugs are important issues to be solved.Hyperpolarization-activated cyclic nucleotide gated cation channels (HCN channels) participate in many advanced brain activities and are closely related to the occurrence and progression of various brain diseases.Among them, the researches on the role and mechanism of HCN channels in substances addiction are gradually gaining attention.Reviewing the researches regarding substances addiction, abnormal function and abnormal expression of HCN channels were observed in many brain regions under the condition of psychoactive substances addiction.However, it has not yet been able to fully understand the mechanism and the behavioral consequences of this change.Therefore, we review the neurobiological mechanisms of HCN channels in substances addiction induced by opioids, cocaine, cannabis, amphetamines, alcohol and tobacco, in order to provide new ideas for the mechanism researches and treatment of substance addiction.

Relationship between phosphodiesterase 4D gene rs966221 single nucleotide polymorphisms and ischemic stroke / 中华老年心脑血管病杂志

Objective To study the relationship between phosphodiesterase 4D (PDE4D) gene rs966221 single nucleotide polymorphisms (SNPs) and ischemic stroke (IS) in Guangxi Zhuang population.Methods One hundred and one IS patients from Guangxi Zhuang Autonomous Region served as IS pgroup and 104 healthy subjects undergoing physical ecamination served as control group in this study.Their PDE4D gene rs966221 SNPs were detected by SNaPshot technique.The genotypes and frequencies of alleles were compared between the two groups and the relationship between PDE4D gene rs966221 SNPs and IS was analyzed.Results No significant difference was found in the GG,GA,AA genotypes and in the frequencies of G and A alleles between the two groups (0.99％ vs 3.85％,29.70％ vs 21.15％,69.31％ vs 75.00％,P＞0.05;15.84％ vs 14.42％,84.16％ vs 85.58％,P＞0.05).Univariate and multivariate logistic regression analysis showed that the PDE4D gene rs966221 SNPs were not related with the risk of IS in dominant AA vs GG+GA,recessive GG vs AA+GA and additive GG vs AA genetic models (P＞0.05).Conclusion The PDE4D gene rs966221 SNPs are not related with IS in Guangxi Zhuang population.

Hyperpolarization activated cyclic nucleotide-gated cation channels involved in central nervous system diseases:research advances / 国际药学研究杂志

Hyperpolarization activated cyclic nucleotide-gated cation (HCN) channels are involved in neuronal rhythm regulation, synaptic activity, membrane resistance and dendritic integration due to their voltage-gated physiological properties. Recent studies have shown that HCN channels play an important role in central nervous system diseases, such as temporal lobe epilepsy, neuropathic pain, learning and memory disorder, substance abuse addiction and other related diseases. In this paper, we summarize the research progress of HCN channels in central nervous system diseases in recent years.

Daily use of phosphodiesterase type 5 inhibitors as prevention for recurrent priapism / Prevenção do priapismo recorrente com a utilização diária de inibidores da fosfodiesterase tipo 5

Summary Objective: The pathogenesis of recurrent priapism is currently being investigated based on the regulation of the phosphodiesterase 5 (PDE5) enzyme. We explored the daily use of PDE5 inhibitors to treat and prevent priapism recurrences. Method: We administered PDE5 inhibitors using a long-term therapeutic regimen in seven men with recurrent priapism, with a mean age of 29.2 years (range 21 to 35 years). Six men (85.7%) had idiopathic priapism recurrences and one man (24.3%) had sickle cell disease-associated priapism recurrences. Tadalafil 5 mg was administered daily. The mean follow-up was 6.6 months (range 3 to 12 months). Results: Daily long-term oral PDE5 inhibitor therapy alleviated priapism recurrences in all patients. Five (71.4%) had no episodes of priapism and two (28.6%) referred decrease in their episodes of priapism. All patients referred improvement in erectile function. Conclusion: These findings suggest the hypothesis that PDE5 dysregulation exerts a pathogenic role for both sickle cell disease-associated priapism and for idiopathic priapism, and that it offers a molecular target for the therapeutic management of priapism. These preliminary observations suggest that continuous long-term oral PDE5 inhibitor therapy may treat and prevent recurrent priapism.

Resumo Objetivo: Uma das teorias propostas para explicar a etiologia do priapismo recorrente está baseada no mecanismo de regulação da fosfodiesterase tipo 5. Estudamos o uso diário dos inibidores de fosfodiesterase tipo 5 no tratamento e na prevenção do priapismo recorrente. Método: Sete homens com diagnóstico de priapismo recorrente, com idade média de 29,5 anos (21 a 35 anos), utilizaram inibidor de fosfodiesterase tipo 5 em dose diária (tadalafila 5 mg/dia) por período prolongado. Seis homens (85,7%) apresentavam priapismo recorrente de etiologia idiopática, e um homem (24,3%), de etiologia associada à anemia falciforme. O seguimento médio foi de 6,6 meses (3 a 12 meses). Resultados: Todos os pacientes se beneficiaram com a utilização de inibidores de fosfodiesterase tipo 5. Cinco (71,4%) não apresentaram nenhum episódio de priapismo e dois (28,6%) relataram diminuição dos episódios. Todos os pacientes relataram melhora da função erétil. Conclusão: Estes achados sugerem que a hipótese do mecanismo de regulação da fosfodiesterase tipo 5 exerce papel importante na patogenia do priapismo recorrente. O uso contínuo e diário de inibidores da fosfodiesterase tipo 5 pode ser uma opção no tratamento do priapismo recorrente.

Role and functions of HCN channels in nervous system with complicated modulations / 中国药理学与毒理学杂志

Hyperpolarization-activated and cyclic nucleotide-gated(HCN)channels which are distributed in a variety of tissues including excitable cells such as heart cells and different neurons have important functions in the control of heart rate and neuronal excitability. Unlike the typical potassium channels and sodium channels,HCN channels can evoke inward currents while the membrane potential is hyper-polarized. Some studies have discovered that HCN channels were important for the nervous system, due to their special electrophysiological features and regulatory effects on the cellular membrane excit-ability.HCN channels are modulated by such factors such as extracellular molecules and intracellular sig-naling cascades, which make their functions quite special in the brain. Given their role, HCN channels seem to be promising targets for specific brain disease as well as chronic pain.In this review,we focus on the roles and functions of HCN channels with their complex modulator factors and neural activity.

Effect of dronedarone on HCN channel mRNA and protein expression in neonatal rat ventricular myocytes / 重庆医学

Objective To explore the effect of dronedaronel on hyperpolarization-activated cyclic-nucleotide-gated(HCN) channel expression by detecting the change of HCN channel mRNA and protein level before and after giving dronedarone in neonatal rat ventricular myocytes.Methods Neonatal rat ventricular myocytes were separated and digested by type Ⅱ collagenase,and then single ventricular myocytes were collected through differential sticking wall separation method.According to the concentrations (0.1,0.5,1.0,5.0,10.0,20.0 μmol/L of dronedaronel for treating myocytes for 48 h) and time(10 μmol/L of dronedaronel for treating myocytes for 1,6,12,24,48 h)the gradient grouping was conducted.The levels of HCN2 and HCN4 channel mRNA and protein level were determined by real-time PCR and Western blot.Results The HCN2 mRNA and HCN4 mRNA expression levels in concentration gradient group and time gradient group were lower than those in the control group(P＜0.05);compared with the control group,the protein level in the 10 umol/L dronedaronel treatment for 12 h group was significantly down-regulated(P＜ 0.01).Conclusion Dronedaronel could inhibit the expression of HCN2/HCN4 channel mRNA and protein,moreover its action shows the concentration dependency and reaches the maximum at 12 h after medication.

Study of Angiogenesis-Related Phosphodiesterase Inhibitors.

Angiogenesis is the process through which new capillaries form from pre-existing capillaries and venules. Its occurrence depends on the migration of vascular endothelial cells which is inhibited by high levels of cAMP. Such levels can be regulated by the degradation caused by the phosphodiesterases (PDEs). Therefore, by inhibiting the action of PDEs it is assumed that angiogenesis can be inhibited with the prevention of migration of these cells. The aim of this study was to evaluate the effect PDE inhibitors on angiogenesis in mice by using nonspecific inhibitors (aminophylline) and selective inhibitors of PDE4 (roflumilast) and PDE5 (sildenafil). BALB/c mice were used as a model; under anesthesia, the mice had a sponge of 0.5 x 0.5 cm introduced into their dorsal subcutaneous tissue; they were then divided into 4 groups and daily gavage treated: 1) control group (n=13) – treated with 0.3 mL of saline solution; 2) aminophylline group (n=16) – 50 mg/kg; 3) roflumilast group (n=14) – 5mg/kg; 3) sildenafil group (n=12) –100 mg/kg. After 7 days, with the animals anesthetized, a blood sample was drawn for hemoglobin (Hb) measurement, the sponge implant was removed, and its content was obtained in 2 mL of saline solution for hemoglobin measurement. Absorbance levels (A), the amount of Hb from the sponge (S) and the total blood Hb concentration levels from each mouse were evaluated. According to the results obtained, we concluded that aminophylline, roflumilast and sildenafil (phosphodiesterase inhibitors) did not cause any alteration in the angiogenesis evaluated by the sponge-implantation method.

Cyclic nucleotide-gated channels and retinal cone photoreceptor cells function / 中华实验眼科杂志

Cyclic nucleotide-gated (CNG) channels are ion channels which are activated by the binding of cyclic guanosine monophosphate (cGMP) or cyclic adenosine monophosphate (cAMP),they play a central role in the signal transduction pathways of vision and olfaction.Six different genes encode CNG protein,containing four A subunits (A1-A4) and two B subunits (B1 and B3).CNGA3 and CNGB3 have been found to be implicated in achromatopsia-associated mutations.Recently,a huge amount of researches showed the good responses to gene therapy in achromatopsia animal models.This article briefly reviewed the physiological roles of CNG channel in retinal cone photoreceptor cells and the recent research achievements of gene therapy in CNG channel-deficient mouse models with achromatopsia.

Effect of hydrogen sulfide on vasorelaxation and content of guanosine 3', 5'-cyclic phosphate in vascular tissue of rats / 中华实用儿科临床杂志

Obgective To observe the effect of hydrogen sulfide (H2S) on vasorelaxation and expression of guanosine 3',5'-cyclic phosphate (cGMP) and activity of cyclic nucleotide phosphodiesterase (PDE) in vascular tissue.Methods H2S donor was provided by sodium bisulfide sodium hydrosulfide.The isolated perfused rat thoracic aorta rings were used to test the relaxation responses to H2S,which recorded by Power Lab system,and the enzyme linked immuno assay was used to detect intracellular cGMP.The activity of PDE was evaluated by using cyclic nucleotide PDE assay kit.Results (1) H2S relaxed the thoracic aorta rings,and the half maximal effective concentration (EC50) for the H2 S relaxation curve,represented by the corresponding concentration of H2 S that achieved 50％ of the maximum relaxation effect,was (1.79 ± 0.31) × 10-5 mol/L.(2)The cGMP content in vascular tissue increased from (22.29 ± 1.59) pmol/L to(41.45 ± 7.49) pmol/L and (31.35 ± 2.56) pmol/L after incubation with 50 μmol/L and 300 μmol/L H2 S,respectively (t =-3.09,t =-2.88;all P ＜ 0.05,n =7-8).(3) cGMP could be lysed into 5'-guanylicacid(5'-GMP) by PDE,which was an important pathway for cGMP degradation.This study showed that PDE activity was decreased in vascular tissue,the 5'-GMP decreased from (0.52 ±0.06) mol/L to (0.25 ±0.06) mol/Land (0.27 ±0.07) mol/L after incubation with 50 μmol/L and 300 μ mol/L H2S,respectively (t =3.21,t =2.58;all P ＜ 0.05,n =7-8).Conclusion The vasorelaxant effects of H2 S might be related to the inhibited activity of PDE and elevated content of cGMP.

Role of spinal HCN channels in dexmedetomidine-produced antinociceptic effect: in vivo and in vitro experiments / 中华麻醉学杂志

Objective To investigate the role of spinal hyperpolarization-activated cyclic nucleotide-gated (HCN) channels in dexmedetomidine-produced antinociceptic effect.Methods In vivo experiment Thirty wild type C57BL/6J mice and 30 HCN1 gene knockout (HCN1-/-) mice, aged 2-3 months, weighing 19-25 g, were randomly divided into 5 groups (n=6 each) using a random number table: control group (group C), and dexmedetomidine 10, 20, 30 and 40 μg/kg groups (Dex10, Dex20,Dex30 and Dex40 groups).In Dex10, Dex20, Dex30 and Dex40 groups, dexmedetomidine 10, 20, 30 and 40 μg/kg were intraperitoneally injected, respectively.The equal volume of normal saline was given in group C.Before dexmedetomidine administration, and at 15, 30, 45, 60, 75, 90, 105 and 120 min after dexmedetomidine administration, tail flick latency to a thermal nociceptive stimulus was measured, and the percentage of the maximum possible effect (MPE％) was calculated.In vitro experiment HCN1 and HCN2 plasmids and green fluorescent plasmids were transfected into HEK293 cells with liposome 2000.At 24-48 h after transfection, HCN1 and HCN2 channel currents were recorded using whole-cell patch clamp technique.HCN channel currents were recorded as baseline value after rupture of membrane.HEK293 cells were perfused with the extracellular fluid containing different concentrations of dexmedetomidine (0.1, 1.0 or 10.0 μmol/L).After the cells were perfused with dexmedetomidine 0.1 μmol/L for 5 min, HCN currents were recorded.The inhibition rate of currents were calculated.After washout, HCN currents were recorded after the cells were perfused with the next concentration for 5 min.The half-maximal activation voltage (V1/2) of HCN channels and the curve slope were recorded.The difference in V1/2 before and after administration (△V1/2) were calculated.Results Compared with group C, MPE％ was significantly increased in Dex10 group-Dex40 group of wild type and HCN1-/-mice (P＜0.05).Compared with Dex30 and Dex40 groups of wild type mice, MPE％ was significantly decreased in Dex30 and Dex40 groups of HCN1-/-mice (P＜0.05).There was no significant difference in MPE％ between Dex10 group and Dex20 group of wild type and HCN1-/-mice (P＞0.05).Compared with the baseline value, the currents and V1/2 of HCN1 and HCN2 channels in HEK293 cells were significantly decreased when the cells were perfused with dexmedetomidine 0.1, 1.0 and 10.0 μmol/L (P＜0.05).Compared with the value when the cells were perfused with dexmedetomidine 0.1 μmol/L, the currents and V1/2 of HCN1 and HCN2 channels in HEK293 cells were significantly decreased, and inhibition rate of currents and △ V1/2were increased when perfused with dexmedetomidine 1.0 and 10.0 μmol/L (P＜0.05).Compared with the value when the cells were perfused with dexmedetomidine 1.0 μmol/L, the currents and V1/2 of HCN1 and HCN2 channels in HEK293 cells were significantly decreased, and inhibition rate of currents and △ V1/2were increased when perfused with dexmedetomidine 10.0 μmol/L (P＜0.05).There was no significant difference in the activation curve slope of HCN1 and HCN2 channel currents in HEK293 cells when the cells were perfused with dexmedetomidine 0.1, 1.0 and 10.0 μmol/L (P＞0.05).Conclusion Dexmedetomidine-produced antinociceptic effect is likely related to the inhibition of spinal HCN channel opening.

Protective effects of adipose?derived stem cells with phosphodiesterase 5 inhibition by lentivirus?mediated stable gene silencing on ischemia?reperfusion injury of renal tubular epithelial cells / 中华肾脏病杂志

Objective To explore the protective effects of adipose - derived stem cells (ADSCs) with phosphodiesterase 5 inhibition by lentivirus-mediated stable gene silencing on the proliferation and apoptosis of renal tubular epithelial cells induced by ischemia-reperfusion injury in vitro. Methods To isolate cultivate and indentify ADSCs from rats. Lentiviral expression vector of carrying PDE5 shRNA gene was transfected into ADSCs, and a negative control group was set up.Western blotting was used to detect PDE5 protein expression levels. ADSCs were co-cultured with NRK-52E in a transwell system, and NRK-52E cells were treated with ischemia/reoxygenation protocol. Edu assay was performed to evaluate the proliferation of NRK cells, flow cytometry to detect the apoptosis of NRK cells, and ELISA to quantify the protein expressions of fibroblast growth factor (FGF) and hepatocyte growth factor (HGF). The expression of E - cadherin and cytokeratin 18 (CK18) was quantified by real time PCR and flow cytometry. Results Western blotting for PDE5 protein indicated a significant reduction of PDE5 protein levels in PDE5 shRNA transduced population. After the treatment of ischemia/reoxygenation in vitro, the proliferative viability and apoptosis of NRK-52E cells co-cultured with ADSCs induced by PDE5 gene inhibition were significantly improved, compared to the normal group (all P<0.05). And the release of HGF, FGF were markedly enhanced (all P<0.05). Moreover, the NRK-52E cells survival, the expression of E-cadherin and CK18 on PDE5 inhibited ADSCs co-cultured with I/R injured NRK cells was significantly increased compared to that in the negative control group (all P<0.05). Conclusion ADSCs preconditioned by inhibition of PDE5 can be a powerful novel approach to improve the survival of renal tubular cells following ischemia-reperfusion injury, and have an obvious tendency to transform epithelial cells.

Comparison of the pacemaker current in canine sino-atrial node cells and the wild type mHCN2 pacemaker current in neonatal rat myocardial cells / 中华老年医学杂志

Objective To investigate the dynamic characteristics of the pacemaker current of canine sino-atrial node cells and compare them with the wild type mHCN2 pacemaker current overexpressed in neonatal rat myocardial cells.Methods Fresh canine sino-atrial node cells were enzymatically isolated in a calcium-free solution containing collagenase and elastase,and the funny current was recorded and compared with the mHCN2 current overexpressed in cultured neonatal rat myocardial cells under the same experimental conditions.Results The canine sinus node cells were elongated,spindle-shaped or polygonal,with well-defined boundaries,and showed spontaneous beating.The elicited pacemaker current was an inward current and its rise in amplitude quickened as the hyperpolarization potential increased.At V ＝-75 mV,the canine sinus atrial node pacemaker current was (-2.1±0.3) pA/pF and had the same activation kinetics as those of the mHCN2 channel current overexpressed in neonatal rat myocardial cells [τact:(728±137) ms vs.(530±65) ms,P＞0.05].Conclusions Within the physiological range,the pacemaker current in canine sino-atrial node cells and the wild type mHCN2 pacemaker current over expressed in neonatal rat myocardial cells have similar activation kinetics.

Relationship of the phosphodiesterase 4D gene rs153031 polymorphism and the susceptibility of unstable angina pectoris / 重庆医学

Objective To investigate the association between the phosphodiesterase 4D(PDE4D) gene rs153031 polymorphism and the susceptibility of unstable angina pectoris(UAP) in Chinese Han population of Changwu region .Methods The PDE4D gene rs153031 polymorphism was genotyped by Taqman probe in 172 UAP patients(UAP group) ,as well as in gender-and-age-matched 220 subjects without coronary heart disease(CHD)(control group) .Results In this crowd ,there was PDE4D gene rs153031 poly-morphism in patients with UAP and in subjects without CHD .Compared with control group ,frequencies of GG ,GA ,AA genotypes and G allele of rs153031 in UAP group showed no statistical differences (P> 0 .05) .Conclusion In Chinese Han population of Changwu region ,PDE4D gene rs153031 polymorphism shows no association with the susceptibility of UAP .

Role of phosphodiesterase 4B in lipopolysaccharide-induced release of inflammatory factors in rat microglias / 中华麻醉学杂志

Objective To evaluate the role of phosphodiesterase 4B (PDE4B) in lipopolysaccharide (LPS)-induced release of inflammatory factors in rat microglias.Methods The pri mary cultured microglial cells were randomly divided into 5 groups (n =24 each) using a random number table:control group (group C),LPS group,vehicle group,mismatch small interfering RNA (siRNA) group and PDE4B-siRNA group.The cells were incubated for 48 h in C and LPS groups.The cells were transfected with lipofectaminTM RNAiMAX 1 μl for 48 h in vehicle group.In mismatch siRNA and PDE4B-siRNA groups,mismatch siRNA 2 μl and PDE4B-siRNA 2 μl were added to 100μl serum-free culture medium (final concentration of siRNA 20 nmol/L),respectively,lipofectaminTM RNAiMAX 1 μl was added simultaneously and the cells were then transfected for 48 h.The expression of PDE4B protein and mRNA was determined by Western blot and real-time PCR,respectively.The cells were cultured for 24 h in serum-free culture medium containing LPS 100 ng/ml in LPS,vehicle,mismatch siRNA and PDF4B-siRNA groups.Then the release of TNF-α and IL-1β was measured by ELISA and the expression of extracellular signalregulated protein kinase (ERK) and phosphorylated ERK (p-ERK) was detected by Western blot.Results Compared with C group,the expression of PDE4B protein and mRNA was significantly down-regulated in group PDE4BsiRNA (P ＜ 0.05),no significant changes were found in LPS,vehicle and mismatch siRNA groups (P ＞ 0.05),and the release of TNF-α and IU1β was increased and the expression of p-ERK was up-regulated in the other four groups (P ＜ 0.05).Compared with LPS group,the release of TNF-α and IL-1β was decreased and the expression of p-ERK was down-regulated in PDE4B-siRNA group,and no significant changes were found in vehicle and mismatch siRNA groups (P ＞ 0.05).There was no significant difference in ERK expression between the five groups (P ＞ 0.05).Conclusion PDF4B can promote LPS-induced release of inflammatory factors in rat microglias and activation of ERK is involved in the mechanism.

Role of expression of phosphodiesterase 4B in spinal cord in inflammatory responses in a rat model of neuropathic pain: relationship with extracellular signal-regulated kinase / 中华麻醉学杂志

Objective To evaluate the role of expression of phosphodiesterase 4B (PDE4B) in the spinal cord in inflammatory responses in a rat model of neuropathic pain and the relationship with extracellular signal-regulated kinase (ERK).Methods Sixty healthy male Sprague-Dawley rats,weighing 180-220 g,in which intrathecal catheters were implanted at L5,6 interspace,were used.The location of catheters was confirmed 6 days later.The rats were randomly divided into 5 groups (n =12 each):sham operation group (group Sham),normal saline (NS) group,vehicle group (group Ⅴ),mismatch siRNA group (group siR-M),and PDE4B-siRNA group (group siR-B).Neuropathic pain was induced by ligation of L5 spinal nerve (SNL).In Sham group,the L5 spinal nerve was only exposed,but not ligated.Immediately after ligation and on 1,3,5,and 7 days after ligation,10 μl NS,10 μl NS,LipofectaminTM RNAiMAX,PDE4B-siRNA (2 μg/10 μl) encapsulated in mismatch siRNA and PDE4B-siRNA (2 μg/10 μl) encapsulated in LipofectaminTM RNAiMAX were injected intrathecally in Sham,NS,V,siR-M,and siR-B groups,respectively.The mechanical pain threshold was measured at 1 day before and 2,4,6 and 8 days after operation.After behavioral testing on 8th day after operation,the rats were sacrificed and the lumbar segment of the spinal cord was removed for determination of PDE4B protein,ERK and phosphor-ERK (p-ERK)expression,and TNF-α,IL-1β and IL-6 levels.Results Compared with Sham group,the mechanical pain threshold was significantly decreased at 2,4,6 and 8 days after operation in NS,V,siR-M and siR-B groups (P ＜0.05),and no significant change was found in the mechanical pain threshold at 2,4,6 and 8 days after operation (P ＞ 0.05) and the expression of p-ERK and PDE4B protein,and levels of TNF-α,IL-1β and IL-6 were increased at 2,4,6 and 8 days after operation in V and siR-M groups (P ＜ 0.05).Compared with NS group,the mechanical pain threshold was significantly increased,and the expression of p-ERK and PDE4B protein and levels of TNF-α,IL-1β and IL-6 were decreased at 2,4,6 and 8 days after operation (P ＜ 0.05),and no significant change was found in the parameters mentioned above in V and siR-M groups (P ＞ 0.05).Conclusion Up-regulation of the expression of PDE4B protein in the spinal cord is involved in the development of neuropathic pain in rats,which may be related to promoted phosphorylation of ERK in the spinal cord and enhanced inflammatory responses.

Advance in recombinant adeno-associated virus mediated gene therapy for color blindness / 中华实验眼科杂志

Color blindness represents a group of vision disorders characterized by lack of ability to distinguish different colors.The inherited color blindness has been regarded as incurable for a long period of time.Recently,adeno-associated virus(AAV) mediated gene therapy has successfully restored cone system vision in animal models with color blindness caused by different gene mutations.These mutations are presented in human color blindness patients.It is predicted that gene therapy will become a novel treatment for these color blindness victims.In addition,a single gene transfer may achieve long-term correction of color deficiency.

ALOX5AP and PDE4D gene polymorphisms and ischemic stroke / 国际脑血管病杂志

Cerebral infarction is a polygenic disease caused by genetic factors and environmental factors.The first discovery in the Icelanders is that the ALOX5AP and PDE4D gene polymorphism may be associated with cerebral infarction.So far,many conclusions of foreign studies are still controversial.This article will summarize the research status and the progress of these two genes.

A Novel Carbamoyloxy Arylalkanoyl Arylpiperazine Compound (SKL-NP) Inhibits Hyperpolarization-Activated Cyclic Nucleotide-Gated (HCN) Channel Currents in Rat Dorsal Root Ganglion Neurons

In this study, we determined mode of action of a novel carbamoyloxy arylalkanoyl arylpiperazine compound (SKL-NP) on hyperpolarization-activated cyclic nucleotide-gated (HCN) channel currents (Ih) that plays important roles in neuropathic pain. In small or medium-sized dorsal root ganglion (DRG) neurons (<40 microm in diameter) exhibiting tonic firing and prominent Ih, SKL-NP inhibited Ih and spike firings in a concentration dependent manner (IC50=7.85 microM). SKL-NP-induced inhibition of Ih was blocked by pretreatment of pertussis toxin (PTX) and N-ethylmaleimide (NEM) as well as 8-Br-cAMP, a membrane permeable cAMP analogue. These results suggest that SKL-NP modulates Ih in indirect manner by the activation of a Gi-protein coupled receptor that decreases intracellular cAMP concentration. Taken together, SKL-NP has the inhibitory effect on HCN channel currents (I h) in DRG neurons of rats.

Autotransplantation of genetically-altered mesenchymal stem cells in treatment of complete heart block in porcine / 第二军医大学学报

Objective: To transplant the autologous mesenchymal stem cells (MSCs) carrying human hyperpolarization activated cyclic nucleoside gated channel 2 (hHCN2) gene into the His-bundle in porcine model of complete heart block (CHB), so as to assess the possibility of establishing autologous biological pacing. Methods: We constructed the recombinant adenovirus containing hHCN2 gene to transfect the porcine MSCs. After autotransplantation into the Hie-bundle in CHB model, the genetically-altered MSCs were tested for their ability to provide a stable and catecholamine-responsive heart rhythm. Histological and immunofluorescence analyses were also performed for the myocardium of the injection site. Results: Recombinant adenovirus pAd. hHCN2 was successfully constructed. Porcine MSCs were transfected by the adenovirus. After autotransplantation, transgenic MSCs significantly enhanced the heart rates in porcine CHB model compared with the control group (P < 0.01), and the cardiac rhythms in the transgenic MSC group were catecholamine responsive. Tissues obtained from the transplanted heart sites showed that the MSCs survived in the myocardium and overexpressed hHCN2 protein. Conclusion: Transplantation of autologous genetically-altered MSCs into the His-bundle in porcine CHB model can serve as a short-term catecholamine-responsive biological pacemaker.

Aspirin attenuates the anti-inflammatory effects of theophylline via inhibition of cAMP production in mice with non-eosinophilic asthma

Theophylline is commonly used to treat severe asthma and chronic obstructive pulmonary disease (COPD) characterized by non-eosinophilic inflammation. Acetyl salicylic acid (ASA) is one of the most widely used medications worldwide, but up to 20% of patients with asthma experience aggravated respiratory symptoms after taking ASA. Here we evaluated the adverse effect of ASA on the therapeutic effect of theophylline in mice with non-eosinophilic asthma. A non-eosinophilic asthma mouse model was induced by airway sensitization with lipopolysaccharide-containing allergen and then challenged with allergen alone. Therapeutic intervention was performed during allergen challenge. Theophylline inhibited lung inflammation partly induced by Th1 immune response. ASA attenuated the beneficial effects of theophylline. However, co-administration of the ASA metabolite salicylic acid (SA) showed no attenuating effect on theophylline treatment. The therapeutic effect of theophylline was associated with increase in cAMP levels, which was blocked by co-treatment of theophylline and ASA. ASA co-treatment also attenuated the anti-inflammatory effects of a specific phosphodiesterase 4 inhibitor. These results demonstrate that ASA reverses anti-inflammatory effects of theophylline, and that ASA exerts its adverse effects through the inhibition of cAMP production. Our data suggest that ASA reverses lung inflammation in patients taking theophylline, although clinical evidence will be needed.