With deeper studies on the endocrine axis of growth hormone-releasing hormone-growth hormone-insulin-like growth factor(GHRH-GH-IGF-1),the correlation between molecular analysis of the growth hormone receptor (GHR) gene,nucleoside acid polymorphism and idiopathic short stature(ISS) have been explored gradually.The GHR gene abnormalities often occurred in extracellular domain of GHR proteins,resulting in dysfunction of intracellular signal transduction of GHR.When the growth hormone couldn't play a role or be insensitive fully,the ISS happened.As reported,GHR single nucleotide polymorphisms(SNP),particularly the GHR-exon3 polymorphisms,have closely correlated with the susceptibility to ISS.GHR gene abnormalities and SNP often have been related to the levels of serum IGF-1 and growth hormone binding protein,and the response of recombinant human growth hormone therapy.The screening candidate genes like GHR gene in ISS,the expression of functional protein and analysis of SNP can improve level of genetic diagnosis,and would be important for clearing the etiology and regulating the clinical treatment of ISS.

Objective·To construct effective shRNA-lentiviral vector targeting rat Eap1 gene and explore the effect of Eap1 on Kiss1 at cellular level. Methods·Four shRNA sequences targeting rat Eap1 mRNA and one negative control sequence were designed and synthesized, and then recombined with lentiviral vectors. After DNA sequencing identification, the shRNA recombinant vectors were co-transfected into 293T cells with pGag/Pol, pRev, pVSV-G to packaging lentiviral particles and the virus titers were determined. NRK-52E cells were transfected with all the four lentiviral-Eap1-shRNAs and the negative control lentivirus, and cells were harvested after 72 h. Real-time PCR and Western blotting was performed to detect the change of the mRNA and protein level of Eap1 gene. The most effective LV-Eap1-shRNA was used to infect NRK-52E cells [non-lentivirus and Eap1-shRNA(-) lentivirus as control groups], and then the level of Kiss1 mRNA expression was detected after infection for 72 h. Results·Seventy-two hours after transfection, the Eap1 mRNA and protein expression between blank group and LV-shRNA(-) group showed no significant difference; while Eap1 expression in all LV-Eapl-shRNA groups reduced significantly in comparing to the control groups. In addition, the lentiviral-Eap1-shRNA4 was the most effective in RNA interference. After blocking Eap1 expression in NRK-52E cells, the Kiss1 mRNA level was significantly increased comparing to the control groups. Conclusion·The effective Eap1-shRNA lentiviral vector is established successfully. The Kiss1 gene expression decreases after blocking Eap1 in NRK-52E cell line.

BACKGROUND: In association with advanced startover of hypothalamus-pituitary-sexual gland axis, the growth of skeleton is accelerated and epiphyseal fusion occurred earlier in puberty precocity children. In clinic,Chinese drug for nourishing yin and reducing fire retards bone age and the advanced fusion in the disease. At the age of sexual development, Chinese drug for benefiting kidney to supplement essence is replaced to promote sexual development and accelerate skeleton growth. The patients with puberty precocity are the same to that in normal puberty in sexual development and bone growing, just being advanced in time. But, it is still unknown the mechanism of Chinese drugs on bone growth and effects on somatostatin (SS) in bone growth.OBJECTIVE: To explore the regulation of kidney-nournishing Chinese drug on periventricular nucleus (PeVN) SS.DESIGN: Completely randomized group division and control experiment were designed.SETTING: Department of Endocrinology, Children's Hospital Affiliated to Shanghai Jiaotong University MATERIALS: The experiment was performed in Institute of Pediatrics of Children's Hospital Affiliated to Shanghai Fudan University from October 2001 to April 2002, in which, 24 SD white rats were employed, of purebred, female and clean grade. 4 groups were randomized, named the control A, the group with drugs of nourishing yin and reducing fire (experiment group No. 1), the control B and group with drugs of nourishing yin and reducing fire + benefiting kidney to supplement essence (experiment group No.2),6 rats in each group.METHODS: ① Control A:physiological saline was applied for gastricperfusion, 5 mL each time, once per day, totally for 1 month. Experiment No. 1 group: Chinese mixture for nourishing yin and reducing fire was applied for gastric perfusion, composed of shengdihuang (Radix Rehmanniae),zhiguiban (Chinemys reevesii.), huangbai (Cortex Phellodendri) and zhimu (Rhizoma Anemarrhenae) etc, 5 mL each time, once per day, totally for 1 month. Control B: physiological saline wasapplied for gastric perfusion,5 mL each time, once per day, totally for 2 months. Experiment No.2 group:Firstly, Chinese mixture for nourishing yin and reducing fire was applied for gastric perfusion, 5 mL each time, once per day, totally for 1 month.And then Chinese mixture for benefiting kidney to supplement essence was applied for breeding, composed of shudihuang (Radix Rehmanniae Preparata), guibanjiao (Colla Carapacis Testudinis), xianlingpi, lujiaojiao (Colla Cornus Cervi) etc., 5 mL each time, once per day, totally for 1 month.② In situ hybridization staining procedure with floating and routine immunohistochemical technique. Medical imaging analysis software was used to assay PeVN SS mRNA, SS immunoreactive neuron counts and absorption (A value) of positive cell area that reflects the gene expression and protein expression. Number of positive neurons of unit area refers to the positive neuron counts in each square millimeter.③ Singer-factoranalysis of variance was used for the comparison of measurement data. F test was adopted for difference comparison among groups.MAIN OUTCOME MEASURES: Effects of Chinese drugs for nourishing yin and reducing fire and benefiting kidney to supplement essence on PeVN SS gene mRNA and protein expressionRESULTS: 24 rats were all in result analysis. ① PeVN SS mRNA positive neuron counts, positive neuron counts of unit area and A value of positive cell area: Those in experiment No.1 group was higher remarkably than the those of control A (P ＜ 0.05); and those in experiment No.2 group were lower remarkably than those of experiment No. 1 group (P ＜ 0.05). ② PeVN SS positive neuron counts, positive neuron counts of unit area and A value of positive cell area: those in experiment No. 1 group: those in experiment No.1 group were higher remarkably than those of control A (P ＜ 0.05) and those in experiment No.2 group were lower remarkably than those of experiment No. 1 group (P ＜ 0.05).CONCLUSION: By controlling hypothalamus SS gene expression and protein expression, Chinese drugs for nourishing yin and reducing fire and benefiting kidney to supplement essence regulate the synthesis and releasing of pituitary growth hormone.

As the right of patient,informed Consent is of great benefit to patients and doctors.It not only improves the service quality of hospitals and doctor-patient relationship,but also strengthens the doctors law consciousness.

The cerebral salt wasting syndrome and syndrome of inappropriate antidiuretic hormone secretion are frequent in hyponatremia in patients with intracranial disorders.There is a major problem about the incidence,diagnosis,and differentiation of cerebral salt wasting syndrome and syndrome of inappropriate secretion of antidiuretic hormone in patients with acute central nervous system disorders.Clinical presentation of cerebral salt wasting syndrome is similar to syndrome of inappropriate antidiuretic hormone secretion,but the therapy is different.So differential diagnosis is essential for appropriate management.This paper differentiate two kinds of diseases from the pathogenesis,diagnosis and treatment.

Humans ; Medicine, Chinese Traditional ; Psoriasis ; classification ; therapy ; Syndrome

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Agents ; adverse effects ; therapeutic use ; Brain Neoplasms ; secondary ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; secondary ; Exanthema ; chemically induced ; Female ; Humans ; Liver Neoplasms ; secondary ; Lung Neoplasms ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Protein Kinase Inhibitors ; adverse effects ; therapeutic use ; Quinazolines ; adverse effects ; therapeutic use ; Remission Induction ; Survival Rate ; Vomiting ; chemically induced

Arginine Vasopressin ; Humans ; Motion Sickness

Objective: A capillary electrophoresis for the determination of berberine hydrochloride and palmatine hydrochloride in Ruyijinhuang Powder(Rhizoma Curcumae Longae, Radix et Rhizome Rhei, Cortex Phellodendri, Rhizoma Atractylodis, etc.) was established. Methods: The berberine hydrochloride and palmatine hydrochloride were determined using benzoic acid as an internal standard its detection wavelength was at 230nm. The optimized condition was achieved with a fused\|silica capillary 100?m?56.5cm, 10mmol?L -1 phosphate buffer(pH=4.80) containing 30% acetonitrile, a constant voltage of 20kv and temperature at 30℃. Results: Calibration curve was constructed in the range of 5?g?mL -1 ～100?g?mL -1 for berberine hydrochloride and palmatine hydrochloride, respectively. The regression equations were Y=0.02964X -0.00771 ( r =0.9998) and Y =0.01092 X -0.02939 ( r =0.9997) for berberine hydrochloride and palmatine hydrochloride, respectively. The average recovery was 98.7% and 102.1%( n =5), respectively. Conclusion: This method is sample, quick and sensitive.

Objective To explore the possible mechanism for KiSSI gene in control gonadotropin - releasing hormone(GnRH)secretion participating in sexual development onset and normal reproduction regulation by investigating the changes in expression of KiSS1,GnRH in hypothalamus and luteinizing hormone(LH),follicle stimulating hormone (FSH),and estradiol(E2 )in serum by using RNA interference mediated by lentivirus - based vectors,after interfering expression of KiSSI. Methods Ninety female Sprague - Dawley rats of 21 days were randomly divided into 3 groups:interference virus group receiving intracerebroventricular injection of KiSSI - microRNA interference lentivirus;lentivi-rus - control group given intracerebroventricular injection of lentivirus without interference effect;9 g/ L saline control group given intracerebroventricular injection of 9 g/ L saline. Ten rats in each group and the animals were sacrificed at 30 - day - old,35 - day - old,45 - day - old,respectively. Then the expressions of KiSSI and GnRH mRNA were detec-ted in the rat hypothalamus with real - time PCR,the LH,FSH,and E2 in serum was examined with chemiluminescence method. Results The levels of KiSSI mRNA in interference virus group were significantly reduced after being infected with recombinant lentivirus compared with those of 9 g/ L saline control group( at 30 d:0. 106 ± 0. 018;at 35 d:0. 218 ± 0. 025;at 45 d:0. 215 ± 0. 033,all P = 0. 000). The level of GnRH mRNA in interference virus group was sig-nificantly reduced after being infected with recombinant lentivirus compared with that of 9 g/ L saline control group(at 30 d:0. 230 ± 0. 040;at 35 d:0. 407 ± 0. 030,all P = 0. 000). The level of LH in interference virus group was lower than the other 2 groups at 35 d[(0. 101 ± 0. 004)IU/ L,P = 0. 467]. The level of FSH in the interference virus group was lower than that of the other 2 groups at 35 d[(0. 235 ± 0. 014)IU/ L,P = 0. 015]. The level of E2 in the interfe-rence virus group was lower than that of the other 2 groups at 35 d and 45 d[at 35 d:(171. 750 ± 11. 050)nmol/ L, P = 0. 000;at 45 d:(192. 310 ± 13. 100)nmol/ L,P = 0. 010]. Conclusions Lentivirus with KiSSI - microRNA can affect the expression of GnRH mRNA and the levels of sex hormone. Lateral cerebral ventricle microinjection of KiSSI -microRNA lentivirus can delay sexual development of Sprague - Dawley female rats.