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According to the latest global cancer statistics， the incidence and mortality of lung cancer rank first in China. Classical therapies remain the most common cancer treatment options， such as surgical resection， radiotherapy， and chemotherapy， but not all cancer patients respond to classical therapies， which require new lung cancer treatment strategies. After decades of research and development， cancer immunotherapy has achieved certain curative effect， which provides new possibilities for cancer treatment. Cyclic guanosine monophosphate-adenosine monophosphate synthase （cGAS） is a cytosolic DNA sensor. It can induce protective immune defense responses against various DNA-containing pathogens and provide anti-tumor immunity by activating the interferon （IFN） gene stimulator （STING） protein. At present， relevant researchers in China and abroad have done a lot of research on the occurrence and development of lung cancer and the pathophysiological mechanism of drug intervention in the treatment of lung cancer. The results show that cGAS/STING signaling pathway plays an important role in the development of the disease， and traditional Chinese medicine monomers or compounds can intervene in lung cancer cells by regulating the cGAS/STING signaling pathway， induce their autophagy and death， regulate their cycle operation， promote senescence， inhibit their proliferation and tumor angiogenesis， promote their invasion and metastasis， and promote the immune activation of anti-lung cancer cells， so as to inhibit or delay the occurrence and development of lung cancer. In recent years， the related research results have been updated rapidly， and the previous literature has not included the latest research results in time， which causes a lot of inconvenience for many scholars to search the literature. Based on this， this paper mainly summarized the mechanism of cGAS/STING signaling pathway intervention in lung cancer in China and abroad in recent years， as well as the research progress of related traditional Chinese medicine intervention， so as to provide new ideas for the development of lung cancer in molecular biology， drug treatment research， and clinical new drug research and provide a reference for further mechanism research.
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Targeting cGAS-STING pathway is a promising strategy in tumor treatment. The pattern recognition receptor cGAS identifies dsDNA and catalyzes the formation of the second messenger 2'3'-cGAMP, activating the downstream interferons and pro-inflammatory cytokines through the adaptor protein STING. Notably, in tumor immune microenvironment, key components of cGAS-STING pathway are transferred among neighboring cells. The intercellular transmission under these contexts serves to sustain and amplify innate immune responses while facilitating the emergence of adaptive immunity. The membrane-based system, including extracellular vesicles transport, phagocytosis and membrane fusion transmit dsDNA, cGAMP and activated STING, enhancing the immune surveillance and inflammatory. The membrane proteins, including specific protein channel and intercellular gap junctions, transfer cGAMP and dsDNA, which are crucial to regulate immune responses. And the ligand-receptor interactions for interferons transmission amplifies the anti-tumor response. This review elaborates on the regulatory mechanisms of cell-to-cell communications of cGAS-STING pathway in tumor immune microenvironment. We further explore how these mechanisms modulate immunological processes and discuss potential interventions and immunotherapeutic strategies targeting these signaling cascades.
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Objective To establish the method for the simultaneous determination of hypoxanthine, inosine, guanosine and adenosine in Dilong formula granules by HPLC and compare the fingerprints of Dilong formula granules from different manufacturers by HPLC chromatogram. Methods The contents of hypoxanthine, inosine, guanosine and adenosine were determined by Thermo AcclaimTM120C18 column (4.6 mm×250 mm 5 μm). The mobile phase was acetonitrile-water. Gradient elution with flow rate of 0.6 ml/min was used. Column temperature was 25 ℃. Detection wavelength was 254 nm. 10 batches of samples were tested. The HPLC chromatogram were compared and analyzed by using the similarity Evaluation system of chromatographic fingerprint of traditional Chinese Medicine (version 2012.130723). Results The linear ranges for the detection of hypoxanthine, inosine, guanosine and adenosine showed good linear relationships within their own ranges (r≥0.999 9). The average recoveries were 99.20%~102.98% with RSD of 0.26 %~0.71%. The contents of 4 components in 10 batches of samples were 0.740 0~4.457 4 mg/g, 2.132 3~7.805 0 mg/g, 0.325 4~1.596 1 mg/g, 0.537 2~2.222 9 mg/g respectively. The similarity between HPLC chromatogram and control fingerprints of Dilong formula granules from different manufacturers was greater than 0.91. Conclusion The method could be used to determine the contents of hypoxanthine, inosine, guanosine and adenosine in Dilong formula granule. HPLC fingerprints could be used to evaluation the quality in Dilong formula granule. The similarity of HPLC fingerprints from different manufacturer production of Dilong formula granule is high, but 4 contents in composition are difference.
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ObjectiveTo observe the therapeutic effect of Yiyuan Qiwei pills （YYQW） on diabetes mellitus-induced induced erectile dysfunction （DMED） in rats and explore its regulation on the nitric oxide （NO）-cyclic guanosine monophosphate （cGMP） signaling pathway. MethodFifty-five healthy SD male rats of clean grade aged 2-3 months underwent intraperitoneal injection of streptozotocin （STZ） to induce the DMED model， and another 10 healthy SD male rats of clean grade aged 2-3 months were assigned to the control group. The model rats were randomly divided into a model group， a sildenafil group （5 mg·kg-1， ig）， and low-， medium-， and high-dose YYQW groups （1.5， 3.0， 6.0 g·kg-1， ig）. The rats in the model group and the control group were given normal saline by gavage at 10 mL·kg-1， once a day for two months. After intervention， the penile erectile function of rats in each group was measured by a pressure detection system. The pathological changes and ultrastructure of penile corpus cavernosum were observed by hematoxylin-eosin （HE） staining and transmission electron microscopy， respectively. The level of NO in the corpus cavernosum was detected by nitrate reductase. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of cGMP and advanced glycation end products （AGEs）. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of endothelial nitric oxide synthase （eNOS）， neurogenic nitric oxide synthase （nNOS）， total nitric oxide synthase （NOS）， and phosphodiesterase type5 （PDE5） in rat penile tissues. The expression of above proteins was detected by Western blot. ResultCompared with the control group， the model group showed decreased intracavernous pressure （ICP）， NO， and cGMP levels， reduced mRNA and protein expression of nNOS and NOS， and increased PDE5 mRNA and protein expression （P<0.05）. Compared with the model group， the sildenafil group and the YYQW groups displayed increased ICP， NO， and cGMP levels， elevated mRNA and protein expression levels of nNOS and NOS， and reduced PDE5 mRNA and protein expression levels （P<0.05）. There were no pathological changes in the tissues and cell ultrastructure of the corpus cavernosum in the control group， while serious pathological changes were observed in the model group. Additionally， the sildenafil group and the YYQW groups were superior to the model group， the optimal effect was observed in the high-dose YYQW group. ConclusionYYQW can improve the penile erectile function of DMED rats and reduce the pathological damage of corpus cavernosum. The mechanism may be related to the promotion of nNOS and NOS expression， the inhibition of PDE5 expression， and the activation of the NO/cGMP signaling pathway.
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ObjectiveTo observe the therapeutic effect of Yiyuan Qiwei pills （YYQW） on diabetes mellitus-induced induced erectile dysfunction （DMED） in rats and explore its regulation on the nitric oxide （NO）-cyclic guanosine monophosphate （cGMP） signaling pathway. MethodFifty-five healthy SD male rats of clean grade aged 2-3 months underwent intraperitoneal injection of streptozotocin （STZ） to induce the DMED model， and another 10 healthy SD male rats of clean grade aged 2-3 months were assigned to the control group. The model rats were randomly divided into a model group， a sildenafil group （5 mg·kg-1， ig）， and low-， medium-， and high-dose YYQW groups （1.5， 3.0， 6.0 g·kg-1， ig）. The rats in the model group and the control group were given normal saline by gavage at 10 mL·kg-1， once a day for two months. After intervention， the penile erectile function of rats in each group was measured by a pressure detection system. The pathological changes and ultrastructure of penile corpus cavernosum were observed by hematoxylin-eosin （HE） staining and transmission electron microscopy， respectively. The level of NO in the corpus cavernosum was detected by nitrate reductase. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of cGMP and advanced glycation end products （AGEs）. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of endothelial nitric oxide synthase （eNOS）， neurogenic nitric oxide synthase （nNOS）， total nitric oxide synthase （NOS）， and phosphodiesterase type5 （PDE5） in rat penile tissues. The expression of above proteins was detected by Western blot. ResultCompared with the control group， the model group showed decreased intracavernous pressure （ICP）， NO， and cGMP levels， reduced mRNA and protein expression of nNOS and NOS， and increased PDE5 mRNA and protein expression （P<0.05）. Compared with the model group， the sildenafil group and the YYQW groups displayed increased ICP， NO， and cGMP levels， elevated mRNA and protein expression levels of nNOS and NOS， and reduced PDE5 mRNA and protein expression levels （P<0.05）. There were no pathological changes in the tissues and cell ultrastructure of the corpus cavernosum in the control group， while serious pathological changes were observed in the model group. Additionally， the sildenafil group and the YYQW groups were superior to the model group， the optimal effect was observed in the high-dose YYQW group. ConclusionYYQW can improve the penile erectile function of DMED rats and reduce the pathological damage of corpus cavernosum. The mechanism may be related to the promotion of nNOS and NOS expression， the inhibition of PDE5 expression， and the activation of the NO/cGMP signaling pathway.
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Mitochondria is one of the important organelles, which is composed of outer mitochondrial membrane and inner mitochondrial membrane. Mitochondrial structure and function are regulated by mitochondrial dynamics. Mitochondrial fusion- and fission-related proteins may participate in the process of mitochondrial fusion and fission, mediate mitochondrial dynamics, thereby regulating cell structure, function and energy metabolism. Mitofusin (MFN) 2, a protein located on the outer mitochondrial membrane of mammalian, has guanosine triphosphatase activity, which may mediate mitochondrial fusion, participate in mitophagy, formation of mitochondria-associated endoplasmic reticulum membrane and apoptosis, and significantly affect the incidence and development of ischemia-reperfusion injury (IRI). In this article, the structure, regulation, function of MFN2 and its role in IRI were reviewed, and the relationship between MFN2 and IRI and underlying mechanism were investigated, aiming to provide novel targets and ideas for the prevention and treatment of IRI.
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An eco-friendly and fast HPLC method was developed for the determination of adenosine, inosine, guanosine and uridine in Cordyceps and related products (fermented mycelia of Hirsutella sinensis andPaecilomyces hepiali). The sample was ultrasonically extracted using 0.5% phosphoric acid solutions for 2.5 min. Sample separation was performed on a Poroshell SB-Aq column (50 mm × 4.6 mm, 2.7 μm) using eco-friendly mobile phase consisting of formic acid and ammonium formate aqueous solution at a flow rate of 1.0 mL·min
Subject(s)
Adenosine , Chromatography, High Pressure Liquid , Cordyceps , NucleosidesABSTRACT
Inflammatory bowel disease is a recurrent chronic intestinal inflammatory disease with unknown etiology and no effective treatment. Phosphodiesterase (PDE) regulates a variety of physiological and pathophysiological processes by mediating the hydrolysis of intracellular second messengers cyclic adenosine monophosphate and cyclic guanosine monophosphate. In recent years, a series of researches suggest that PDE inhibitors such as several PDE4 inhibitors, PDE5 inhibitors (sildenafil, tadalafil and vardenafil), PDE3 inhibitors (cilostazol), PDE9 inhibitor (PF-04447943) and PDE3/PDE4 double inhibitor (pumafentrine) have ameliorating effect on experimental colitis in animals. In clinical trials, PDE4 inhibitor apremilast showed more therapeutic advantage than tetomilast. This article reviews the recent research progress of PDE inhibitors in treatment of inflammatory bowel disease.
Subject(s)
Animals , Colitis , Inflammatory Bowel Diseases/drug therapy , Phosphodiesterase 4 InhibitorsABSTRACT
Depression was a complex and difficult to regulate disease, which was closely related to purinergic system and purine metabolism disorder. Although there had been studies to improve depression by regulating purinergic system, the mechanism of action was complex and needed to be sorted out. Recently, a large number of studies had found that the addition of exogenous purine metabolites adenosine, inosine and guanosine had a significant antidepressant effect, indicating that regulating the level of purine substances in purine metabolism could also improve depression, which was of great significance to the further study of the pathogenesis and treatment of depression. In view of this, this study reviewed the relationship between purinergic system or purine metabolism and depression, in order to provide a reference for the further study of the pathogenesis of depression.
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Propolis from stingless bees (Heterotrigona itama) is a resinous compound that exhibits antihyperglycaemia, free radical scavenging, and cardioprotective properties. The effect of propolis on diabetic vessels has not been investigated. Thus, this research aimed to determine the effect of propolis supplementation on the level of antioxidants and its mechanism of action in the aorta of diabetic rats. Male Sprague-Dawley rats were divided into five groups (n=8/group): healthy (control), untreated diabetes (DM), metformin-treated diabetes (DM+M, 300 mg/kg/day metformin), propolis-treated diabetes (DM+P, 300 mg/kg/day propolis extract) and diabetes with combined treatment (DM+M+P, dosage as former). Oral supplementation was conducted for four weeks immediately upon successful induction of diabetes by streptozotocin (60 mg/kg, intraperitoneal injection). At the end of the study, the rats were euthanised, and thoracic aorta was processed into tissue homogenates to determine the levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase-1 (GPx-1) and soluble receptor for advanced glycation end-products (sRAGE). Aorta segments were harvested to examine their relaxation response towards graded concentration of acetylcholine (Ach; 10-8-10-4) M following precontraction with phenylephrine (PE; 10-6 M). Vasorelaxation towards a cumulative dose of propolis (0.01-1.00%) using PE-precontracted healthy aorta (n=6/experiments) was investigated under various simulated conditions: physiological buffer, L-NAME (10-4 M), methylene blue (10-5 M), indomethacin (10-5 M) and elevated glucose (25 mM). Propolis maintained antioxidative enzymes and sRAGE decoy molecules in the aortic tissue of the diabetic rats. The amelioration of diabetes-induced impairment of endothelium-dependent relaxation by propolis was mediated through the nitric oxide(NO)-cyclic guanosine monophosphate (cGMP) pathway. This non-clinical study reports vasoprotective property of propolis in diabetes mellitus.
Subject(s)
Animals , Male , Rats , Propolis/analysis , Bees/anatomy & histology , Rats, Sprague-Dawley/classification , Diabetes Mellitus/drug therapy , Endothelium/abnormalities , Nitric Oxide/adverse effects , Aorta/abnormalities , Relaxation , Vasodilation , Antioxidants/pharmacologyABSTRACT
Background and Purpose: Hie-symptom is more common in women, with complains of strong cold sensation of fingers and lower limbs during cold weather. From the cyanotic findings of hands and thighs and dark venous blood, blood stasis due to excessive peripheral vein contraction was suspected. Then we studied the changes of sublingual and body surface temperature, venous gas partial pressure in the warm and cold conditions. To examine the role of thermo-dilating effects of nitric oxide (NO), the effects oral administration PDE 5 inhibitor Tadarafil (TDF) were also studied. Subjects and Methods: The subjects were 10 women (31 +- 8.8 yrs) with Hie-symptom and 7 women (26+-3.7 yrs) without Hie-symptom, BMI, blood pressure, heart rate,sublingual and peripheral body surface temperature (hand and lower limb), venous and arterial blood gas partial pressure, and fingertip arterial oxygen saturation were measured. The measurement was carried out at warm indoors (about 23°C) and cold outdoors (about 12°C). Then 10 mg TDF tablet was taken and all measurements were repeated again at the same time on the next day. Results: There was no difference in fingertip arterial blood oxygen saturation in both groups either at indoor or outdoor conditions, and even after taking TDF. In the cold outdoor, the subjects with Hie-symptom, compared to without Hie-symptom, showed significantly lower body surface temperature and venous blood pO2, and increased pCO2. After taking TDF, although sublingual temperature and the decrease in body surface temperature outside the room improved in both groups, the improvement was greater in Hie-symptom. Consideration and Conclusion: Because of normal fingertip arterial blood oxygen saturation, Hie-symptom is not considered to be a disorder of the cardiopulmonary/arterial system. From a significant decrease in peripheral body surface temperature, and peripheral venous blood pO2, and an increase in pCO2 of Hie-symptom in cold outdoors, it is considered that blood stasis by excessive constriction of peripheral veins or arteriovenous anastomosis (AVA) by the cold. The better effects of oral TDF, in Hie-symptom seems to predict the involvement of NO or cGMP in blood stasis.
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Objective::To improve the quality control system in the production of Jinshuibao capsules, and to provide experimental basis for the follow-up research and application of this preparation. Method::High performance liquid chromatography (HPLC) was employed, the analysis was performed on a Ultimate AQ-C18 column (4.6 mm×150 mm, 5 μm). The chromatographic conditions for the determination of adenosine, guanosine and uridine were as following: mobile phase of methanol-0.1%formic acid aqueous solution for gradient elution, the flow rate of 0.4 mL·min-1, column temperature at 30 ℃, sample quantity of 10 μL, detection wavelength at 260 nm. The chromatographic conditions for the determination of ergosterol were as follows: mobile phase of methanol-water (98∶2), the flow rate of 1 mL·min-1, column temperature at 25 ℃, sample quantity of 10 μL, detection wavelength at 283 nm. Result::The main chromatographic peaks of fermented Cordyceps powder samples in different production stages showed little difference. The linear relationships of adenosine, guanosine and uridine were good (R2 >0.999), their recoveries were 106.06%, 101.25%and 105.88%, respectively. The contents of adenosine and ergosterol in 20 batches of samples extracted from 2016 to 2018 were in line with the requirements in the 2015 edition of Chinese Pharmacopoeia, the contents of guanosine and uridine were 0.97-1.36, 0.67-1.38 mg/capsule, respectively. Conclusion::The quality of Jinshuibao capsules in the market is stable. This method can be used to detect the quality of Jinshuibao capsules, and it is simple, stable and reliable.
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Vav-family proteins are guanosine nucleotide exchange factors (GEFs) and one of RhoGEFs family numbers.Vav proteins can influence cell signaling transduction,cytoskeleton recombination and adherence migration in special cells by RhoA,RhoG and Rac-1 pathway.There are 80 kinds of RhoGEFs in mammalian cell.Vav proteins play an important role in hematological system,nervous system,cardiovascular system and immune system.So the function of vav protein in immune system was reviewed in this paper.
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OBJECTIVE@#To evaluate the therapeutic effect of the combined treatment with balance acupuncture therapy and exercise re-learning rehabilitation therapy and the impact on serum cAMP and cGMP in the patients with hemiplegia of cerebral ischemic stroke.@*METHODS@#A total of 90 patients of hemiplegia of cerebral ischemic stroke were randomized into an observation group and a control group, 45 cases in each one. All of the patients in the two groups received health education, diet guidance, routine symptomatic treatment as well as exercise re-learning rehabilitation therapy. Additionally, in the observation group, balance acupuncture therapy was applied, in which, the acupoints on the aspect of the human body, on the governor vessel and bladder meridian were adopted in the morning and those on the aspect of the human body, on the conception vessel and kidney meridian were stimulated in the afternoon. In the control group, the regular acupuncture was given. In the two groups, both acupuncture and rehabilitation therapies were given 5 days a week, 2 week-treatment as one course and totally 2 courses were required. Separately, before and after treatment, the score of Fugl-Meyer assessment (FMA) and the score of Chinese stroke scale (CSS) were recorded, the levels of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) detected in serum and the clinical therapeutic effect were evaluated in the two groups.@*RESULTS@#After treatment, FMA score was increased in the patients of either of the groups as compared with that before treatment (<0.01) and CSS score decreased as compared with that before treatment (<0.01). After treatment, FMA score in the observation group was higher than that in the control group (<0.01) and CSS score was lower than the control group (<0.01). After treatment, the level of serum cAMP of the patients in either of the groups was increased as compared with that before treatment (<0.01) and that of cGMP decreased as compared with that before treatment (<0.01). After treatment, the level of cAMP in the observation group was higher than that in the control group (<0.01) and that of cGMP was lower than the control group (<0.01). The total effective rate was 93.3% (42/45) in the observation group, better than 73.3% (33/45) in the control group (<0.01).@*CONCLUSION@#The balance acupuncture therapy combined with exercise re-learning rehabilitation effectively improves the motor function of the affected limb, relieves injury and regulate the levels of serum cAMP and cGMP in the patients with hemiplegia of ischemic stroke.
Subject(s)
Humans , Acupuncture Points , Acupuncture Therapy , Brain Ischemia , Therapeutics , Cyclic AMP , Blood , Cyclic GMP , Blood , Hemiplegia , Therapeutics , Stroke , Therapeutics , Stroke Rehabilitation , Treatment OutcomeABSTRACT
Background and Purpose: Hie-symptom is more common in women, with complains of strong cold sensation of fingers and lower limbs during cold weather. From the cyanotic findings of hands and thighs and dark venous blood, blood stasis due to excessive peripheral vein contraction was suspected. Then we studied the changes of sublingual and body surface temperature, venous gas partial pressure in the warm and cold conditions. To examine the role of thermo-dilating effects of nitric oxide (NO), the effects oral administration PDE 5 inhibitor Tadarafil (TDF) were also studied. Subjects and Methods: The subjects were 10 women (31 +- 8.8 yrs) with Hie-symptom and 7 women (26+-3.7 yrs) without Hie-symptom, BMI, blood pressure, heart rate,sublingual and peripheral body surface temperature (hand and lower limb), venous and arterial blood gas partial pressure, and fingertip arterial oxygen saturation were measured. The measurement was carried out at warm indoors (about 23°C) and cold outdoors (about 12°C). Then 10 mg TDF tablet was taken and all measurements were repeated again at the same time on the next day. Results: There was no difference in fingertip arterial blood oxygen saturation in both groups either at indoor or outdoor conditions, and even after taking TDF. In the cold outdoor, the subjects with Hie-symptom, compared to without Hie-symptom, showed significantly lower body surface temperature and venous blood pO2, and increased pCO2. After taking TDF, although sublingual temperature and the decrease in body surface temperature outside the room improved in both groups, the improvement was greater in Hie-symptom. Consideration and Conclusion: Because of normal fingertip arterial blood oxygen saturation, Hie-symptom is not considered to be a disorder of the cardiopulmonary/arterial system. From a significant decrease in peripheral body surface temperature, and peripheral venous blood pO2, and an increase in pCO2 of Hie-symptom in cold outdoors, it is considered that blood stasis by excessive constriction of peripheral veins or arteriovenous anastomosis (AVA) by the cold. The better effects of oral TDF, in Hie-symptom seems to predict the involvement of NO or cGMP in blood stasis.
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OBJECTIVE@#To test the effect of Banxia Xiexin Decoction (, BXD) on the contraction and relaxation of gastric smooth muscle (SM) in diabetic gastroparesis (DGP) model rats, and to explore the mechanism of BXD in the prevention and treatment of DGP through experiments of signal pathway both in vivo and in vitro.@*METHODS@#Sixty Sprague-Dawley rats were divided into 6 groups according to a random number table: control group, model group, high-, medium- and low-dose BXD groups (9.2, 4.6 and 1.8 g/(kg·d), respectively), and domperidone group (10 mg/(kg·d)), 10 rats per group. DGP model was established initially by a single intraperitoneal injection of streptozotocin (STZ), and was confirmed by recording gastric emptying, intestinal transport velocity and gastric myoelectric activity of rats after 2 months. Each group was treated with a corresponding drug for 4 weeks. The mRNA and protein expressions of phospholipase C (PLC), inositol triphosphate (IP@*RESULTS@#Compared with the model group, high- and medium-dose BXD and domperidone significantly increased the expressions of PLC, IP@*CONCLUSIONS@#Treatment with high- and medium-dose BXD significantly attenuated STZ-induced experimental DGP in rats. The therapeutic effect of BXD on DGP rats might be associated with the PLC-IP
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Vav-family proteins are guanosine nucleotide exchange factors(GEFs)and one of RhoGEFs family numbers.Vav proteins can influence cell signaling transduction, cytoskeleton recombination and adherence migration in special cells by RhoA, RhoG and Rac-1 pathway.There are 80 kinds of RhoGEFs in mammalian cell.Vav proteins play an important role in hematological system, nervous system, cardiovascular system and immune system.So the function of vav protein in immune system was reviewed in this paper.
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@#【Objective】 To study the mutation characteristics of Tyrosine hydroxyls（TH） gene in a pedigree with dopa-responsive dystonia（DRD）. 【Methods】 Extraction of genomic DNA from peripheral blood of a proband and his parents and two sisters using high- throughput sequencing （NGS） method were detected on 256 known pathogenicity genes associated with dystonia and dyskinesia.【Results】Mutations on tyrosine hydroxylase（TH）gene in the exon 14 and exon 9 were detected in the proband and his eldest sister in this pedigree. They had a complex heterozygosity of c.1481C > T（p.Thr494Met）and c.943G >A（p.Gly315Ser），and one heterozygous mutation was carried by parents respectively. The mutation was not detected in his second sister and 50 people with normal phenotype controls. 【Conclusion】 The mutations of TH gene c. 1481C > T（p.Thr494Met）and c. 943G > A（p.Gly315Ser）led to the gene abnormality in DRD family，and a new mutation of TH gene was found，which expanded the relationship between DRD genotype and clinical phenotype. It is vital that early accurate diagnosis and treatment of DRD is the key to improve prognosis.
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Objective To establish a UPLC method for the simultaneous determination of uracil, adenosine, guanosine, and uridine in Aconiti Lateralis Radix Praeparata (ALRP) and Aconiti Radix (AR), and to compare the content difference of four nucleosides in the samples based on multivariate statistical analysis. Methods Nucleosides in ALRP and AR were extracted with purified water by ultrasonic extraction. The UPLC method was performed on a BEH C18 (100 mm × 2.1 mm, 1.7 μm) through a gradient elution of acetonitrile and water at a flow rate of 0.2 mL/min with column temperature at 30 ℃. The injection volume was set at 4.0 μL, and the detection wavelength was set at 260 nm. Difference significance analysis, hierarchical cluster heatmap analysis, principal component analysis, and TOPSIS analysis were used for data processing to comprehensively evaluate the quality of ALRP and AR. Results The method was in accordance with the regulations, the quantitative evaluation of uracil, adenosine, guanosine, and uridine was in good linear range (r2 > 0.999 3), and the average recovery was 99.86%, 99.14%, 99.74%, and 98.71% respectively, and the RSDs were all less than 2.0%. Taking the four nucleosides as indexes, the samples No. 18 of ALRP and the samples No. 23 of AR were the best in quality. Conclusion The established method is simple, accurate, and reliable with good precision, repeatability, and stability, which can be used for the simultaneous determination of four nucleosides in ALRP and AR, and it might provide the scientific basis for the study of water-soluble constituents in ALRP and AR.
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<p><b>OBJECTIVE</b>To investigate whether the methanol extract of Berberis amurensis Rupr. (BAR) augments penile erection using in vitro and in vivo experiments.</p><p><b>METHODS</b>The ex vivo study used corpus cavernosum strips prepared from adult male New Zealand White rabbits. In in vivo studies for intracavernous pressure (ICP), blood pressure, mean arterial pressure (MAP), and increase of peak ICP were continuously monitored during electrical stimulation of Sprague-Dawley rats.</p><p><b>RESULTS</b>Preconstricted with phenylephrine (PE) in isolated endotheliumintact rabbit corus cavernosum, BAR relaxed penile smooth muscle in a dose-dependent manner, which was inhibited by pretreatment with NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, and H-[1,2,4]-oxadiazole-[4,3-α]-quinoxalin-1-one, a soluble guanylyl cclase inhibitor. BAR significantly relaxed penile smooth muscles dose-dependently in ex vivo, and this was inhibited by pretreatment with L-NAME H-[1,2,4]-oxadiazole-[4,3-α]-quinoxalin-1-one. BAR-induced relaxation was significantly attenuated by pretreatment with tetraethylammonium (TEA, P<0.01), a nonselective K channel blocker, 4-aminopyridine (4-AP, P<0.01), a voltage-dependent K channel blocker, and charybdotoxin (P<0.01), a large and intermediate conductance Ca sensitive-K channel blocker, respectively. BAR induced an increase in peak ICP, ICP/MAP ratio and area under the curve dose dependently.</p><p><b>CONCLUSION</b>BAR augments penile erection via the nitric oxide/cyclic guanosine monophosphate system and Ca sensitive-K (BK and IK) channels in the corpus cavernosum.</p>