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Objective: To establish a high performance liquid chromatography(HPLC)method for the determi- nation of drug content and related substances in cyproheptadine hydrochloride injection. Methods: COSMOSIL C8 col- umn(4.6 mm×250 mm,5 μm)was used for the determination of drug content. The mobile phase was acetonitrile-phos- phate buffer(0.045 mol/L,adjusted to pH 4.5 with phosphoric acid)(42:58,V/V). The flow rate was 1.0 ml/min. Injec- tion volume was 20 μl. The detection wavelength was 286 nm. By reference to Chinese Pharmacopoeia 2015,the deter- mination of related substances was also carried out by the HPLC method under the same conditions as the drug content determination except for the gradient elution and the detection wavelength(set at 230). Using these established HPLC methods,the drug content was determined for the three batches of self-made cyproheptadine hydrochloride injection samples and the stability of the injection was also investigated. Results: The calibration curve of cyproheptadine hydrochlo- ride was linear in the range of 10-40 μg/ml(Y=40.551X+13.869,r=0.9999). The average recovery was 99.00%(RSD= 0.39%). Under the HPLC condition,cyproheptadine hydrochloride and its related substances were completely separated and the excipients did not interfere with their determination. Conclusion: The established HPLC method is simple,ac- curate,sensitive and specific,which could be used for the determination of drug content and related substances in the cy- proheptadine hydrochloride injection.
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Cyproheptadine is a H1 and 5- HT1/2 receptor antagonists, impairing the anticonvulsant activity of antiepileptic drugs and reduces threshold, increases severity of seizures, when administered chronically. Anuj a 13 years old male from Delhi, known case of SSPE stage-III with epilepsy, on oral anti-epileptic drug has seizure induction followed the use of oral cyproheptadine. Stopping cyproheptadine, patient didn’t sustain any further seizures.
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Cyproheptadine (CPH), a first-generation antihistamine, enhances the delayed rectifier outward K current (I) in mouse cortical neurons through a sigma-1 receptor-mediated protein kinase A pathway. In this study, we aimed to determine the effects of CPH on neuronal excitability in current-clamped pyramidal neurons in mouse medial prefrontal cortex slices. CPH (10 µmol/L) significantly reduced the current density required to generate action potentials (APs) and increased the instantaneous frequency evoked by a depolarizing current. CPH also depolarized the resting membrane potential (RMP), decreased the delay time to elicit an AP, and reduced the spike threshold potential. This effect of CPH was mimicked by a sigma-1 receptor agonist and eliminated by an antagonist. Application of tetraethylammonium (TEA) to block I channels hyperpolarized the RMP and reduced the instantaneous frequency of APs. TEA eliminated the effects of CPH on AP frequency and delay time, but had no effect on spike threshold or RMP. The current-voltage relationship showed that CPH increased the membrane depolarization in response to positive current pulses and hyperpolarization in response to negative current pulses, suggesting that other types of membrane ion channels might also be affected by CPH. These results suggest that CPH increases the excitability of medial prefrontal cortex neurons by regulating TEA-sensitive I channels as well as other TEA-insensitive K channels, probably I and inward-rectifier Kir channels. This effect of CPH may explain its apparent clinical efficacy as an antidepressant and antipsychotic.
Subject(s)
Animals , Female , Cyproheptadine , Pharmacology , Histamine H1 Antagonists , Pharmacology , Membrane Potentials , Physiology , Mice, Inbred C57BL , Patch-Clamp Techniques , Potassium Channel Blockers , Pharmacology , Potassium Channels , Metabolism , Prefrontal Cortex , Physiology , Pyramidal Cells , Physiology , Receptors, sigma , Metabolism , Tetraethylammonium , Pharmacology , Tissue Culture TechniquesABSTRACT
Objective To establish a HPLC method for the assay of cyproheptadine hydrochloride and the related sub-stances.Methods TheHPLCwasperformedonanAgilentEclipseXDB-C18column(4.6mm×250mm,5μm)atthetemper-ature of 25 ℃ with the mobile phase of acetonitrile-buffer Solution ( Dissolve 2 .16 g of sodium octane-1-sulfonate with about 500 ml of water and mix well .Add 10 .0 ml of glacial acetic acid and 5 .0 ml of triethylamine ,and dilute with water to 1 000 ml ,mix well and adjust to pH 7 .0 with triethylamine) (85∶15 ,V/V ) .The flow rate was 1 .0 ml/min and detection wavelength was 286 nm .The injection volume was 10μl .Results The calibration curves of cyproheptadine hydrochloride ,im-purity A ,B and C showed good linear response in the range from 0 .056 2 to 5 .620 μg/ml (r= 0 .999 8) ,0 .052 4 to 5 .240 μg/ml(r=1 .000 0) ,0 .050 3 to 5 .032 μg/ml (r=0 .999 9) and 0 .053 2 to 5 .316 μg/ml(r=0 .999 8) respectively .The LOQs for cyproheptadine hydrochloride ,impurity A ,B and C were within 0 .049-0 .054 μg/ml ,LODs were within 0 .019-0 .022 μg/ml with recovery between 98%-100% .RSD of repeatability was 5 .5% (n=6) .Conclusion This method gave an accurate and reliable results .It can be used for quality control of cyproheptadine hydrochloride .
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Objective:To establish a determination method for the content of cyproheptadine hydyochloride tablets and the related substances in the tablets by HPLC. Methods:The assay was performed on a CAPCELL PAK C18(Shiseido)(250 mm ×4.6 mm, 5μm) column with methanol-0. 002 5mol·L-1 sodium heptanesulfonate (adjusting pH to 3 with phosphoric acid)(60: 40) as the mo-bile phase. The detection wavelength was 225 nm, the flow rate was 1. 0 ml·min-1 , the column temperature was 30℃ and the sample size was 10 μl. Results: Cyproheptadine hydyochloride had good linear relationship within the range of 4. 12-82. 40 μg·ml-1 ( r=1. 000 0), and the average recovery was 99. 2%(RSD=0. 8%, n=9). The peaks of the related substances were well separated from that of cyproheptadine hydrochloride. Conclusion:The method is simple, fast and accurate, and can be used for the quality control of cyproheptadine hydyochloride tablets.
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Using cyproheptadine ( CYP) as template molecule, methacrylic acid ( MAA) as monomer, ethylene glycol dimethacrylate (EGDMA) as cross-linker, molecularly imprinted polymers (MIP) with high selectivity to cyproheptadine (CYP) were prepared by the optimization of porogen, monomer, and the mole ratio of monomer to template. The specific surface area of the prepared polymers was 24. 9 m2 / g. The recovery of CYP was above 94. 0% when the following procedure was applied to the cartridge of MIP as adsorptive material: conditioning with methanol and water, loading with water, washing with water and methanol, and eluting with methanol-ammonia (95: 5, V/ V). As a control, the recovery of CYP on non-imprinted polymers cartridge (NISPE) was only 38. 9% . The binding capacity of the molecularly imprinted solid phase extraction (MISPE) towards CYP found to be about 8. 8 mg of CYP/ g polymers and the imprinting factor (IF) was about 2. 32. Under optimal conditions, a mixed standard solution of CYP, amitriptyline, sulfadiazine and trimethoprim (10 mg / L each) was uploaded on the MISPE and NISPE for selectivity experiment. The gradient elution was used by using 0. 05% sodium pentanesulfonate solution (A)-acetintrile (B) as a mobile phase. The recoveries on the MISPE for sulfadiazine and trimethoprim (different structure with CYP) were less than 10% , however, the recovery for the similar structural amitriptyline was more than 70% , and the recovery more than 90% for CYP. All the recoveries on the NISPE for four analytes were less than 30% . This new MISPE cartridge was applied to extract and enrich CYP in livestock drinking water sample, and the recoveries of CYP ranged from 80. 5% -97. 7% , and the limit of detection (LOD) was 0. 01 mg / L.
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The goal of the present investigation was to design and evaluate mucoadhesive buccal patches of Cyproheptadine Hydrochloride (CPH) which is a sedating antihistamine with antimuscarinic, serotonin-antagonist, and calcium-channel blocking action. Buccal films were made with Hydroxy propylcellulose (HPC EF) and Hydroxy Propyl Methyl Cellulose (HPMC E15) as mucoadhesive polymers. Permeation of CPH was calculated ex vivo using porcine buccal membrane. The patches were evaluated for weight variation, thickness variation, surface pH, moisture absorption, in vitro residence time, mechanical properties, in vitro release, ex vivo permeation studies and drug content uniformity. The formulation F8 of HPMC E15 was found to give the better results and release of drug from the film followed Higuchi and Korsmeyer and Peppas models.
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OBJETIVO: O objetivo deste estudo foi determinar se a administração de ciproheptadina é capaz de induzir ganho de peso em pacientes com fibrose cística. MÉTODOS: Foi realizado um estudo duplo-cego, controlado com placebo em dois centros no Brasil. Vinte e cinco pacientes com fibrose cística entre 5 e 18 anos completaram o estudo. Os pacientes foram randomizados em dois grupos, para receber ciproheptadina 4 mg três vezes por dia durante 12 semanas ou placebo. Todos os dados foram coletados no início e no final do período de estudo e incluíram peso, altura e espirometria. RESULTADOS: O ganho de peso médio foi de 0,67 kg e 1,61 kg nos grupos placebo e ciproheptadina, respectivamente (p = 0,036). O índice de massa corporal (IMC) diminuiu 0,07 kg/m² no grupo placebo e aumentou 0,46 kg/m² no grupo intervenção (p = 0,027). A mudança no IMC para a idade (escore z) foi de -0,19 no grupo placebo e 0,20 no grupo ciproheptadina (p = 0,003). O IMC escore z diminuiu 0,19 no grupo placebo e aumentou 0,2 no grupo ciproheptadina (p = 0,003). Alterações na função pulmonar não foram estatisticamente diferentes. CONCLUSÃO: O uso de ciproheptadina em pacientes com fibrose cística foi bem tolerado, apresentando um ganho de peso significativo e um aumento no IMC após 12 semanas. Foi encontrado um tamanho de efeito clinicamente relevante para o peso/idade (escore z) e IMC para idade (escore z). Tais achados sugerem que a prescrição de ciproheptadina pode ser uma abordagem alternativa para pacientes que precisam de suporte nutricional por um curto período de tempo.
OBJECTIVE: To determine whether the administration of cyproheptadine was able to induce weight gain in patients with cystic fibrosis. METHODS: We performed a double-blind, placebo-controlled trial in two centers in Brazil. Twenty-five patients with cystic fibrosis between 5 and 18 years completed the study. Patients were randomized into two groups, to receive either cyproheptadine 4 mg three times per day for 12 weeks or placebo. All data were collected at the beginning and at the end of the study period and included weight, height and spirometry. RESULTS: Average weight gain was 0.67 kg in the placebo group and 1.61 kg in the cyproheptadine group (p = 0.036). Body mass index (BMI) decreased 0.07 kg/m² in the placebo group and increased 0.46 kg/m² in the intervention group (p = 0,027). The change in BMI for age (z score) was -0.19 in the placebo group and +0.20 in the cyproheptadine group (p = 0.003). BMI z score decreased 0.19 in the placebo group and increased 0.2 in the cyproheptadine group (p = 0.003). Changes in pulmonary function were not statistically different. CONCLUSION: Use of cyproheptadine in cystic fibrosis patients was well tolerated, showing a significant weight gain and a significant increase in BMI after 12 weeks. A clinically relevant effect size for weight/age (z score) and body mass index for age (z score) was found. Such findings suggest that the prescription of cyproheptadine can be an alternative approach for patients who need nutritional support for a short period of time.
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Appetite Stimulants/therapeutic use , Body Mass Index , Cyproheptadine/therapeutic use , Cystic Fibrosis/complications , Weight Gain/drug effects , Double-Blind Method , SpirometryABSTRACT
Aims: To study anticonvulsant and central nervous system depressant activity of methanol leaf extract of Croton zambesicus (MECZ) in Swiss albino mice and investigate the role of serotonin in these activities. Methodology: Anticonvulsant activity of graded doses (200, 300 and 400 mg/kg p.o) of MECZ was assessed through seizures induced by picrotoxin and pentylenetetrazole (PTZ). Effects of the extract on pentobarbitone-induced sleep and amphetamine-induced stereotype behavior were also evaluated. Possible involvement of serotonergic pathways was studied using cyproheptadine (4mg/kg i.p), a non-selective serotonin antagonist (5-HT1/5HT2). Results: In both picrotoxin and PTZ-induced seizures, the extract significantly delayed onset of seizure (p<0.05) in a dose-dependent manner and provided significant protection against death. There was a dose-dependent increase of pentobarbitone sleeping time and a significant reduction (p<0.05) in the sleep latency. The extract also produced a significant reduction in amphetamine-induced stereotype behavior. Pretreatment with cyproheptadine abolished the anticonvulsant effect of the extract. The inhibitory effect of the extract on amphetamine-induced hyperactivity and its potentiation of pentobarbitone-induced sleep were also reversed by cyproheptadine. Conclusion: The results of this study showed that methanol extract of Croton zambesicus leaf possesses anticonvulsant activity and other CNS depressant activities and these activities are possibly mediated through interaction between serotonergic and GABAergic transmissions.
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El síndrome serotoninérgico (SS) se caracteriza por la presencia de disfunción autonómica, deterioro neuromuscular y alteraciones en el estado mental. El diagnóstico es clínico y por laboratorio. El tratamiento es mediante medidas de apoyo a base de líquidos, hipotermia, benzodiacepinas y, cuando lo amerite, intubación y ventilación mecánica. El pilar de la intervención farmacológica es ciproheptadina, un antagonista central H1, con un antagonismo periférico 5-HT2A. El medicamento disponible sólo se puede administrar por vía oral, por lo que en enfermos críticos debe administrarse a través de sonda nasoyeyunal. El objetivo de este trabajo es presentar un caso de SS y revisar la literatura al respecto.
The serotonin syndrome (SS) is characterized by a spectrum of signs characterized by autonomic dysfunction, neuromuscular impairment, and alterations in mental status. The diagnosis of the serotonin syndrome is clinical and by laboratory evaluation. Treatment is with active cooling and benzodiazepines to control agitation, and intubation, especially in patients with severe hyperthermia and delirium. The mainstay of pharmacologic intervention is cyproheptadine, a centrally acting H1-antagonist, with a prominent peripheral 5-HT2A antagonism. The drug is only available orally, and if the patient is unable to swallow, the drug should be administered nasogastrically. The objective of this paper is report a case of Serotonin Syndrome and review the literature related to this disease.
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El síndrome serotonínico es un cuadro neurológico agudo debido a hiperactividad serotoninérgica, por la interacción de drogas que refuerzan o mimetizan la acción del neurotrasmisor. La incidencia del síndrome de serotonina es ascendente por la disponibilidad creciente de fármacos serotoninérgicos como los antidepresivos. Por ello es importante que los médicos reconozcan y manejen adecuadamente el síndrome serotonínico. Este reporte de caso se refiere a una intoxicación accidental por el neuroléptico atípico olanzapina en un niño de 2 años, quien desarrolló manifestaciones clínicas como agitación, sudoración, mioclonías, clonus espontáneo e hipertermia, considerados como criterios diagnóstico del cuadro. La terapia consistió en descontaminación interna con lavado gástrico, carbón activado y sulfato de sodio, ciproheptadina, propranolol y furosemida. Su evolución fue satisfactoria. En nuestro país hay disponibilidad de la mayoría de los fármacos causales y tienen amplio uso, por lo que es probable el subregistro del síndrome. De allí la importancia de este reporte de caso
Serotonin syndrome is an acute neurologic picture due to serotonergic hyperactivity, due to the interaction of drugs that enhance or mimic the action of the serotonin. The incidence of serotonin syndrome is rising because of the growing availability of serotonergic drugs such as antidepressants. It is therefore important that clinicians recognize and manage appropriately this syndrome. This case report refers to an accidental poisoning by the atypical neuroleptic olanzapine in a 2 year old boy who developed clinical manifestations such as agitation, sweating, myoclonus, spontaneous clonus and hyperthermia, considered as diagnostic criteria for the syndrome. Therapy consisted of internal decontamination with gastric lavage, activated charcoal and sodium sulfate, cyproheptadine, propranolol and furosemide. The clinical outcome was satisfactory. In our country the majority of the causal drugs are easily available and widely employed, for which reason it is probable that this syndrome is under registered. Hence the importance of this case report
Subject(s)
Humans , Male , Child, Preschool , Cyproheptadine/therapeutic use , Poisoning/complications , Serotonin Agents/adverse effects , Serotonin Syndrome/diagnosis , Serotonin Syndrome/therapy , PediatricsABSTRACT
OBJECTIVE:To establish a method for content determination of cyproheptadine hydrochloride in fuyan cream.METHODS:Ultraviolet spectophotometry was applied to determine the contents of cyprohepatadine hydrochloride as well as its adjuvant as two coexisting components without isolation and extraction,the wavelengths were286nm and258nm res_ pectively,linear regression and simultaneous equations were used to analyze the results.RESULTS:Cyproheptadine hydrochlor_ ide and its adjuvant showed good linear relationship in the range of2.52～25.2and2.5022～25.022?g/ml respectively,the average recovery of cyproheptadine hydrochloride was98.61%(RSD=1.08%).CONCLUSION:The present method is simple,convenient,reproducible and reliable,which is suitable for rapid determination for fuyan cream.
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AIM To study whether cyproheptadine(Cyp) affects endocrine functions in reproductive system with gender difference. METHODS Sixty SD rats were randomly distributed into 3 groups according to gender, respectively, and they were administered NS(5 mL*kg-1*d-1), Cyp 2.4, 4.8 mg*kg-1*d-1 accordingly by ig for 14 d or 21 d. The serum levels of luteinizing hormone(LH), follicle stimulating hormone(FSH), testosterone(T), progesterone(P), estrodiol(E2) were measured by radio-immunoassay and the ultrastructure of gonadotropin-releasing hormone(GnRH) cells, gonadotropin cells, Leydig cells, Sertoli cells, luteal cells, granulocytes and so on were observed by electronmicroscopy and microscopy. The calmodulin(CaM) mRNA expression in hypothalamus-pituitary-testis axis(HPTA) was detected by reverse transcriptase-polymerase chain reaction. RESULTS Cyp(2.4 mg*kg-1*d-1×14 d, ig) increased serum LH concentration while decreased serum FSH, P concentrations in female rats. Cyp(4.8 mg*kg-1*d-1×14 d, ig)increased serum LH, T concentrations in males, and increased serum LH concentration while decreased serum FSH, E2 and P concentrations significantly in females. The retrograde changes of ultrastructure were observed in part of gonadotropin and ovary endocrine cells, while a stimulating one in testis endocrine cells. CaM mRNA expression levels were elevated in testis but not in hypothalamus and pituitary in male rats. CONCLUSIONCyp had a negative effect on endocrine function in females, but a positive one in males. The ultrastructure showed relevant changes in target gland. Cyp promoted CaM mRNA expression in testis,which had close connections with Cyp′s stimulative effect in HPTA.
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AIM: To study the effects of cyproheptadine (Cyp) and anisodamine (Ani)on the changes of intracellular free Ca 2+ concentration ([Ca 2+ ] i) induced by tumor necrosis factor (TNF ?) in single endothelial cells, and to explore the mechanisms of TNF ?-mediated shock and antishock actions of Cyp and Ani. METHODS: Human umbilical vein endothelial cell strains(ECV304) were seed in 35 mm tissue culture dish with 2 mL DMEM culture medium. The cultured cells were loaded by Fluo-3/AM. The spatial distribution and the dynamic changes of [Ca 2+ ] i in single endothelial cell was determined by laser scanning confocal microscopy(LSCM). RESULTS: [Ca 2+ ] i in single endothelial cell after stimulation of TNF ? rapidly increased in a dose-dependent manner and approached the peak value within 60 seconds, afterwards, decreased and kept above the basal level. The confocal scanning image showed that [Ca 2+ ] i elevation was more obvious in nuclear than in cytoplasma, and decreased slowly. Cyp (3?10 -5 , 6?10 -5 mol/L) and Ani (2?10 -5 , 4?10 -5 mol?L -1 ) markedly inhibited TNF ? (1.2?10 -9 mol?L -1 )-induced [Ca 2+ ] i elevation. CONCLUSIONS: TNF ? markedly induces elevation of [Ca 2+ ] i in single endothelial cell, it may be an important mechanism of TNF ?-induced shock and tissue injury. Cyp and Ani obviously suppress TNF ?-induced [Ca 2+ ] i elevation, which probably is one of the mechanisms of their antishock effects.
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Objective To study the potentiation of chloroquine activity and mechanism by ketotifen and cyproheptadine in in vitro cultured Plasmodium falciparum Fcc SM1/yN strain. Methods In vitro cultured Fcc SM1/yN strain was added to pre-prepared drug plates at 50 ?l/well after synchronization to make final concentration of 0.312 5-2 560 nmol/L for chloroqine and of 9.80-5 000 nmol/L for ketotifen or cyproheptadine. After 34 hours' culture in 37 ℃, the number of schizonts with 3 or more nuclei was calculated among 200 parasites under microscope. Calculated half inhibitive concentration ( IC50 ) of chloroquine and every drug combination to parasite as well as chloroquine activity enhancement index ( AEI) of ketotifen (or cyproheptadin) . Time dependency of potentiation was studied. All data were analyzed statistically with SPSS 13.0. After 20 hours' action of one optimal combination dose of chloroquine/ketotifen or chloroquine/cyproheptadine, RNA of the Fcc SM1/yN strain was extracted and real-time PCR was used to determine the expression level of pfcrt and pfmdr1 genes. Results The best potentiation effect was observed with ketotifen or cyproheptadine of 625 nmol/L, with IC50 of 74.53 nmol/L for chloroquine/ketotifen and 89.7 nmol/L for chloroquine/cyproheptadine respectively, and activity enhancement index (AEI) of 0.42 for chloroquine/ketotifen and 0.30 for chloroquine/cyproheptadine respectively. Combination of 625 nmol/Lketotifen or cyproheptadine with 5 nmol/L chloroquine showed the highest potentiation potency. 6-7 hours during which ketotifen or cyproheptadine was added after chloroquine showed the highest effect, with IC50 of 67.70 nmol/L for chloroquine/ketotifen and 81.53 nmol/L for chloroquine/cyproheptadine respectively, and the AEI was 0.47 for chloroquine/ketotifen and 0.37 for chloroquine/cyproheptadine respectively. After action of chloroquine/ketotifen or chloroquine/ cyproheptadine at one optimal combination dose, expression level of pfcrt gene increased by 91% and that of pfmdr1 gene decreased by 14% respectively. Conclusion Appropriate combination of chloroquine/ketotiphen or chloroquine/ cyproheptadine potentiates chloroquine against in vitro cultured P. falciparum. 6-7 hour period is an optimal time when ketotifen or cyproheptadine was added after chloroquine. Potentiating activity of ketotifen and cyproheptadine may be related to the expression level of pfcr t and pfmdr1 genes.
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OBJECTIVE: Neuroleptic Induced Akathisia(NIA) often occurs in neuroleptic treated patients. Cyproheptadine, an antiserotonergic agent, was used to treat neuroleptic induced akathisia. METHOD: In an open trial 21 neuroleptic-treated patients with akathisia were administrated Cyproheptadine(16mg/day) over 4 days. Assessment of akathisia was evaluated using Barnes' rating scale(BAS) for neuroleptic induced akathisia. The degree of depression and psychosis were assessed by brief psychiatric rating scale(BFRS) and Hamilton rating scale for depression(HAM-D). RESULT: Most patients(20 of 21) with neuroleptic induced akathisia(NIA) showed improvement under the treatment of cyproheptadine. There was no aggravation of psychosis or depression during the treatment. Symptoms of akathisia aggravated when cyproheptadine was discontinued. CONCLUSION: Cyproheptadine may be useful in the treatment of neuroleptic induced akathisia(NIA).
Subject(s)
Humans , Cyproheptadine , Depression , Psychomotor Agitation , Psychotic DisordersABSTRACT
Scrum glucose, growth hormone and cortisol were determined at 1 a.m. and 7 a.m. in 40 patients with diabetes mel-litus. 18 cases were treated with cyproheptadine. Effect of cyproheptadine on "dawn phenomenon" of diabetes mel-litus was observed. The results showed that serum glucose, growth hormone and cortisol were higher at 7 a.m than at 1 a.m. Cyproheptadine could significantly decrease 7 a.m. serum glucose, growth hormone and cortisol. Those data suggested that diabetic dawn phenomenon has concern with the increase of growth hormone and cortisol level and cyproheptadine can be used for preventing and treating this dawn phenomenon.
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AIM To study the effects of cyproheptadine (Cyp) on endocrine functions and pituitary adrenalcortex axid in rats and its mechanism METHODS Radioimmunoassay(RIA)was used to measure the serum levels of ACTH and cortisone Ultramicrostructure of the ACTH cells and adrenalcortex and the calmodulin(CaM) mRNA expression of the pituitary and adrenalcortex were observed by electron microscope and the quantity PCR technique RESULTS Cyp 2 3 and 4 6 mg?kg -1 ?d -1 , ig for 14 d had significantly reduced the serum levels of ACTH and cortisone ( P