OBJECTIVE: To evaluate the relationship between the tyrosine hydroxylase (TH) gene and mental disorder by comparing the two TH gene SNP points (extron 3 and intron 9) of patients with mental disorders and normal human. METHODS: DNA extracted from specimens of patients and normal subjects were quantified by using Real-time PCR. RESULTS: (1) G334A distribution was: G=0.133, A=0.867 in patients with schizophrenia, G=0.116, A=0.884 in patients with depression, and G=0.214, A=0.786 in normal group. (2) C5162G distribution was: C=0.962, G=0.038 in patients with schizophrenia, C=0.959, G=0.041 in patients with depression, and C=0.961, G=0.039 in normal group. CONCLUSION There are statistically significant differences in G334A locus between normal people and patients with mental disorders, but no statistically significant differences in C5162G locus.
Depressive Disorder/genetics* ; Female ; Forensic Genetics ; Humans ; Male ; Mental Disorders/genetics* ; Polymorphism, Single Nucleotide/genetics* ; Schizophrenia/genetics* ; Tyrosine 3-Monooxygenase/genetics*

Neurochemical investigation has played a major role in the search for the cause of schizophrenia. Among many hypotheses, dopamine hypothesis of schizophrenia prevails despite much criticism and qualification. Recently, evidences showing the atypical antipsychotics act via serotonergic mechanism suggest serotonin system as an etiologic factor for schizophrenia. We examined the possibility of the association of enzymes critical for the synthesis of serotonin (tryptophan hydroxylase, TPH) and dopamine (tyrosine hydroxylase, TH) with schizophrenia. The regions of DNA that has been known to be polymorphic were amplified using polymerase chain reaction from the peripheral blood cells of 374 biologically unrelated schizophrenic patients and 393 healthy controls. RFLP (A218C) and VNTR polymorphism (intron 1) were examined for TPH and TH, respectively. The patterns of polymorphisms and the frequencies of each allele were not significantly different between the control and the patient groups, suggesting no possible associations of the genetic polymorphisms of TPH and TH genes and schizophrenia. However, in schizophrenics, the frequency of A type allele was significantly higher in positive group than negative group. Thess findings suggest the association of positive schizophrenia with A type allele of TH gene.
Alleles ; Antipsychotic Agents ; Blood Cells ; DNA ; Dopamine ; Genetics ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Schizophrenia* ; Serotonin ; Tryptophan Hydroxylase* ; Tryptophan* ; Tyrosine 3-Monooxygenase* ; Tyrosine*

Until recently, the etiology of schizophrenia was generally attributed to abnormalities in dopaminergic neurotransmission. Specifically, an excess of dopaminergic activity in the mesolimbic system has been postulated to produce the positive symptoms, while decreased dopaminergic activity in the mesocortical system has been suggested to cause negative symptoms. Accordingly, we performed an association study of schizophrenia with TH gene. Three hundred and seventy four biologically unrelated schizophrenic patients meeting DSM-III-R criteria from Kangnam St. Mary's Hospital affiliated with Catholic university of Korea were recruited for our study. The 393 healthy controls were volunteers for DNA library of Kangnam St. Mary's Hospital without personal or family history of psychiatric and neurologic illness. DNA was extracted from peripheral mononuclear cells and polymorphic region was amplified by polymerase chain reaction. TH intron 1 VNTR polymorphism was typed by silver staining. The allele distributions of TH gene were not different between schizophrenics and controls. However, the frequency of allele A was significantly higher in positive group than that of negative group of schizophrenics. These findings suggest that poitive schizophrenia may be associated with allele A of TH gene.
Alleles ; DNA ; Gene Library ; Genetics ; Humans ; Introns* ; Korea ; Polymerase Chain Reaction ; Schizophrenia ; Silver Staining ; Synaptic Transmission ; Tyrosine 3-Monooxygenase* ; Tyrosine* ; Volunteers

OBJECTIVETo investigate the potential of gene therapy of rat prolactinomas mediated by adenoviral vectors with a gene encoding rat tyrosine hydroxylase.

METHODSRecombinant replication-deficient adenovirus named Ad-GFP-TH with rat TH-cDNA and control adenovirus named Ad-GFP were constructed by homologous recombination in bacterial cells. The rat pituitary prolactinoma cell line MMQ are chosen as the target cells to study the effect of gene therapy on their growth and prolactin secretion mediated by Ad-GFP-TH.

RESULTSRecombinant Ad-GFP-TH and Ad-GFP were successfully reconstructed. Transfection of MMQ cells with Ad-GFP-TH not only restrained their growth but also decreased their PRL secretion.

CONCLUSIONGene therapy may serve for a potential treatment for prolactinomas, especially invasive prolactinomas.

Adenoviridae ; genetics ; Animals ; Genetic Therapy ; Genetic Vectors ; Pituitary Neoplasms ; therapy ; Prolactinoma ; therapy ; Rats ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Tyrosine 3-Monooxygenase ; biosynthesis ; genetics

OBJECTIVE: To investigate the association of tyrosine hydroxylase (TH) C-824T polymorphism with essential hypertension (EH) susceptibility in Hunan Han population by a case-control study. METHODS: A case-control study was performed on 368 EH patients and 353 healthy controls of Han nationality recruited in Hunan province. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)was used to genotype the C-824T polymorphism. RESULTS: (1) Genotype frequencies for TH-824CC and -824CT+-824TT genotypes were 89.9% and 10.1%, respectively for EH patients and 88.7% and 11.3%, respectively for the controls. No significant difference in the genotype distribution of C-824T polymorphism between the patients and controls was observed (P=0.579). Allele frequencies of TH C-824T also showed no significant difference between the patients and controls (P=0.515). (2) When adjusted by EH risk factors, results of unconditional logistic regression analysis showed that there was no association between TH C-824T polymorphism and EH susceptibility (P=0.264). (3) When stratified by gender, no significant difference in the genotype distribution of TH C-824T polymorphism was observed between the patients and controls in either male or female subjects (P=0.841 and P=0.288). (4) Diastolic blood pressure (DBP) in individuals with -824 CT+TT genotype was significantly higher than that in individuals with -824 CC genotype in the controls (P=0.015). (5) When stratified by gender, significant difference in DBP between TH C-824T CT+TT genotype and CC genotype was observed in the male (P= 0.018) but not in the female (P=0.083) controls. CONCLUSION There is no association between TH gene C-824T polymorphism and EH susceptibility in Hunan Han population. The TH gene C-824T polymorphism is possibly associated with increased DBP in the males in Hunan Han population.
Adult ; Case-Control Studies ; China ; ethnology ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Hypertension ; enzymology ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Sex Factors ; Tyrosine 3-Monooxygenase ; genetics

Objective: To explore the clinical and genetic characteristics of children with dopa-responsive dystonia (DRD) caused by tyrosine hydroxylase (TH) gene variations. Methods: Clinical data of 9 children with DRD caused by TH gene variations diagnosed in the Department of Children Rehabilitation, the Third Affiliated Hospital of Zhengzhou University from January 2017 to August 2022 were retrospectively collected and analyzed, including the general conditions, clinical manifestations, laboratory tests, gene variations and follow-up data. Results: Of the 9 children with DRD caused by TH gene variations, 3 were males and 6 were females. The age at diagnosis was 12.0 (8.0, 15.0) months. The initial symptoms of the 8 severe patients were motor delay or degression. Clinical symptoms of the severe patients included motor delay (8 cases), truncal hypotonia (8 cases), limb muscle hypotonia (7 cases), hypokinesia (6 cases), decreased facial expression (4 cases), tremor (3 cases), limb dystonia (3 cases), diurnal fluctuation (2 cases), ptosis (2 cases), limb muscle hypertonia (1 case) and drooling (1 case). The initial symptom of the very severe patient was motor delay. Clinical symptoms of the very severe patient included motor delay, truncal hypotonia, oculogyric crises, status dystonicus, hypokinesia, decreased facial expression, and decreased sleep. Eleven TH gene variants were found, including 5 missense variants, 3 splice site variants, 2 nonsense variants, and 1 insertion variant, as well as 2 novel variants (c.941C>A (p.T314K), c.316_317insCGT (p.F106delinsSF)). Nine patients were followed up for 40 (29, 43) months, and no one was lost to follow-up. Seven of the 8 severe patients were treated by levodopa and benserazide hydrochloride tablets and 1 severe patient was treated by levodopa tablets. All the severe patients responded well to levodopa and benserazide hydrochloride tablets or levodopa tablets. Although the weight of the patients increased and the drug dosage was not increased, the curative effect remained stable and there was no obvious adverse reaction. One severe patient developed dyskinesia in the early stage of treatment with levodopa and benserazide hydrochloride tablets and it disappeared after oral administration of benzhexol hydrochloride tablets. Until the last follow-up, motor development of 7 severe patients returned to normal and 1 severe patient still had motor delay due to receiving levodopa and benserazide hydrochloride tablets for only 2 months. The very severe patient was extremely sensitive to levodopa and benserazide hydrochloride tablets and no improvement was observed in this patient. Conclusions: Most of the DRD caused by TH gene variations are severe form. The clinical manifestations are varied and easily misdiagnosed. Patients of the severe patients responded well to levodopa and benserazide hydrochloride tablets or levodopa tablets, and it takes a long time before full effects of treatment become established. Long-term effect is stable without increasing the drug dosage, and no obvious side effect is observed.
Female ; Humans ; Infant ; Male ; Benserazide/therapeutic use* ; Dystonia/genetics* ; Hypokinesia/drug therapy* ; Levodopa/pharmacology* ; Muscle Hypotonia ; Retrospective Studies ; Tyrosine 3-Monooxygenase/genetics*

OBJECTIVETo construct a recombinant adenovirus for carry tyrosine hydroxylase (TH) gene and expressing bioactive TH protein in the animal model of Parkinson disease.

METHODSThe TH gene was inserted into the shuttle plasmid, which was transformed into E.coli BJ-5183 for homologous recombination with the adenovirus genome. 293 cells were transfected with the recombinant adenovirus genome to obtain the recombinant virus, and the transcription and expression of TH were determined by RT-PCR and immunofluorescence assay, respectively. The production of L-DOPA in the in vitro reaction system was determined using capillary electrophoresis.

RESULTSWe have successfully constructed the recombinant adenovirus. The TH mRNA and the corresponding protein were detected by RT-PCR and immunofluoresence assay in 293 cells. L-DOPA was also detected in the reaction system.

CONCLUSIONThe adenovirus constructed allows efficient expression of bioactive TH protein in vitro, which provides a basis for future study of gene therapy of Parkinson disease in animal models.

Adenoviridae ; genetics ; metabolism ; Cell Line ; Electrophoresis, Capillary ; Escherichia coli ; genetics ; metabolism ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Levodopa ; analysis ; biosynthesis ; genetics ; Parkinson Disease ; therapy ; Recombinant Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Tyrosine 3-Monooxygenase ; biosynthesis ; genetics

OBJECTIVETo explore the mutation of tyrosine hydroxylase(TH) gene in Chinese patients with autosomal recessive(AR) dopa-responsive dystonia(DRD) and to lay a solid basis for gene diagnosis of AR-DRD in China.

METHODSMutation analysis of TH gene was performed in 5 probands with AR-DRD and 2 sporadic patients with DRD by use of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combining DNA direct sequencing.

RESULTSThe PCR-SSCP analysis and DNA direct sequencing following PCR revealed no mutation in all the 14 exons of TH gene.

CONCLUSIONThe mutation rate of TH gene in Chinese patients with AR-DRD is low, hence suggesting the genetic heterogeneity and a new locus for AR-DRD.

Asian Continental Ancestry Group ; genetics ; China ; Dystonia ; ethnology ; genetics ; Female ; Genes, Recessive ; genetics ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA ; Tyrosine 3-Monooxygenase ; genetics

In the experiment, we designed and synthesized two siRNAs based on the sequence of nuclear receptor-related factor 1 (Nurr1) mRNA. They were separately subcloned into the plasmid of pSilenCircle (pSC) containing U6 promoter. The pSC-Nurr1 vectors (pSC-N1 and pSC-N2) specific to Nurr1 gene and the negative control vector of short-hairpin RNA (shRNA) eukaryotic expression vector were constructed. We cultured the dopaminergic cell line MN9D and the verified vectors were transfected with LipofectamineTM 2000 in vitro. The positive cell clones transfected with pSC were obtained after being screened with 500 mug/ml G418. After that, the silencing effects of Nurr1 and TH mRNA or protein were detected by real time RT-PCR and Western blot. The neurite extension of MN9D cells was observed and photographed by inverted microscope. The results showed that Nurr1 mRNA expression in MN9D cells was specifically down-regulated by the vectors of pSC-N1 and pSC-N2, and the silencing effects were 62.3% and 45.6%, respectively. The dopaminergic phenotype of TH mRNA was also suppressed significantly and the silencing effects were 76.3% and 62.6%, respectively. Meanwhile, the expressions of Nurr1 and TH proteins were also significantly suppressed, and the silencing effects of Nurr1 and TH protein were 57.4%, 72.0% and 79.1%, 70.1% respectively. The negative control and liposome groups had no effect on the two genes. In conclusion, Nurr1 shRNA expressing vectors can inhibit the expressions of Nurr1 and TH mRNA or protein in MN9D cells, and Nurr1 might play a role in neurite extension of MN9D cells. Nurr1 shRNA expressing vector may provide a novel applicable strategy for the study on the function of the genes associated with Parkinson disease and the development of dopaminergic neuron.
Cell Line ; Dopaminergic Neurons ; cytology ; metabolism ; Down-Regulation ; Fetus ; Humans ; Mesencephalon ; cytology ; Neurites ; physiology ; Nuclear Receptor Subfamily 4, Group A, Member 2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Tyrosine 3-Monooxygenase ; genetics ; metabolism

OBJECTIVETo explore the role of inflammatory reaction mediated by p38-mitogen activated protein kinase (p38-MAPK) signal path on prevention and treatment of Parkinson disease (PD) model rats by electroacupuncture (EA).

METHODSThirty-two healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group and an EA group, eight rats in each one. The PD model was established in the model group and EA group by subcutaneous injection of rotenone in skin-back area (2 mg/kg, dissolved in sunflower oil, 2 mg/mL in density), while the injection of sunflower oil emulsion without rotenone at the same point and quantity as the model group was applied in the sham operation group. The normal group was not given any intervention. The EA treatment (continuous wave, 2 Hz in frequency, 1 mA in intensity, 20 min) was applied at "Fengfu" (GV 16) and "Taichong" (LR 3) in the EA group, once a day for continuously 14 days. No treatment was given in the other groups. The expression of tyrosine hydroxylase (TH), phosphorylated p38-MAPK, cyclooxygenase-2 (COX-2) in the substantia nigra were detected with immunohistochemical method.

RESULTSThere was typical PD ethology change in the model group. Compared with the normal group and sham operation group, the expression of TH positive neuron in the substantia nigra in the model group was significantly decreased, while the expression of phosphorylated p38-MAPK and COX-2 were significantly increased (all P < 0.01). Compared with the model group, the expression of TH positive neuron in the EA group was apparently increased, while the expression of phosphorylated p38-MAPK and COX-2 were significantly decreased (all P < 0.01).

CONCLUSIONThe EA therapy could obviously reduce the expression of inflammation mediator COX-2, inhibit the phosphorylation of p38-MAPK, reduce the damage of dopaminergic neurons in the rats with PD, and this effect may be related with the impact of p38-MAPK signal path

Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Electroacupuncture ; Humans ; Male ; Parkinson Disease ; enzymology ; genetics ; therapy ; Phosphorylation ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; enzymology ; Tyrosine 3-Monooxygenase ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism