Objective · To investigate effect of fucoidan on autophagy, migration and invasion in human multiple myeloma U266 cells. Methods · The U266 cells treated with fucoidan were cultured in vitro. The formation of autophagosomes was observed by transmission electron microscopy (TEM). Transwell assay was used to evaluate the effect of fucoidan on migratory and invasive abilities of U266 cells. The protein levels of LC3-Ⅱ/LC3-Ⅰ, Beclin-1, P62, MMP9, CXCR4, p-AKT/T-AKT, p-mTOR/T-mTOR were detected by Western blotting. MMP9 concentration in the culture medium was examined by ELISA. Results · ① Autophagosomes increased in fucoidan-treated cells compared with control group under TEM. ② Migratory and invasive abilities were inhibited by fucoidan in a dose-dependent manner, which were suppressed by chloroquine. ③ Western blotting demonstrated that expression of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and MMP9 increased in fucoidan-treated cells, while P62, CXCR4, p-AKT/T-AKT and p-mTOR/T-mTOR decreased compared with control group. ④ The result of ELISA showed that MMP9 concentration in the culture medium of fucoidan-treated cells significantly decreased. Conclusion · Fucoidan induces autophagy and inhibits migration and invasion in U266 cells.

Objective To investigate the cerebral blood flow (CBF) perfusion patterns and related factors in hyperthyroidism patients.Methods Twenty-five patients with hyperthyroidism and twenty-two healthy controls matched for age,sex,education were enrolled.~(99)Tc~m-ethylene cysteinate dimer (ECD) SPECT CBF perfusion imaging was performed at rest.Statistical parametric mapping 5.0 software (SPM5) was used and a statistical threshold of P＜0.05 (corrected) was applied for signifying changes of regional CBF (rCBF).The semiquantitative values of rCBF were extracted automatically by brain search 1.1 software and were correlated with concentrations of serum thyroid hormones(FT_3,FT_4),thyroid autoimmune antibodies:sensitive thyroid stimulating hormone(sTSH),thyroid peroxidase antibody (TPOAb) and TSH receptor antibody (TRAb) by Pearson analysis,with disease duration by Spearman analysis.Results rCBF was decreased significantly in limbic system and frontal lobe,including parahippocampal gyrus,uncus (posterior entorhinal cortex,posterior parolfactory cortex,parahippocampal cortex,anterior cingulate,right inferior temporal gyrus),left hypothalamus and caudate nucleus (P＜0.05,corrected).rCBF in left lingual gyrus,posterior cingulated was negatively correlated with concentration of FT_3(r=-0.468,-0.417,both P＜0.05).rCBF in left lingual gyrus,bilateral inferior temporal gyrus,right superior parietal lobe was negatively correlated with concentration of FT_4(r=-0.4M,-0.418,-0.415,-0.459,all P＜0.05),while that in left mammillary body and putamen was positively correlated with concentration of FT_4(r=0.419,0.412,both P＜0.05).rCBF in left insula was negatively correlated with concentration of sTSH,and right auditory associated cortex was positively correlated with concentration of sTSH(r=-0.504,0.429,both P＜0.05).rCBF in left middle temporal gyrus,left angular gyrus was positively correlated with concentration of TRAb while that in right thalamus,right hypothalamus,left anterior nucleus,left ventralis nucleus was negatively correlated with concentration of TRAb(r=0.750,0.862,-0.691,-0.835,-0.713,-0.759,all P＜0.05).rCBF in right anterior cingulate,right cuneus,right rectus gyrus,right superior marginal gyrus was positively correlated with concentration of TPOAb(r=0.696,0.581,0.779,0.683,all P＜0.05).rCBF in postcentral gyrus,temporal gyrus,left superior marginal gyrus and auditory associated cortex was positively correlated with disease duration(r=0.502,0.457,0.524,0.440,all P＜0.05).Conclusion Hypoperfusions in limbic system and fontal lobe were found in hyperthyroidism Patients,which might be associated with thyroid function and disesse duration.

Objective To investigate the influence of EMR and ESD of endoscopic surgery on perioperative clinical parameters, complete resection rate and complications of patients with neuroendocrine tumors of digestive tract. Methods 40 patients with neuroendocrine tumors of digestive tract were chosen from June 2009 to June 2016 and randomly divided into 2 groups: A group (20 patients) with EMR and B group (20 patients) with ESD; and the operation time, the treatment cost, the lesion size, the lesion thickness, the complete resection of tumor, the negative rate of vertical margin and the complication incidence of the two groups were compared. Results The operation time and the treatment cost of B group were significant better than A group (P < 0.05). There was no significant difference in the lesion size and the lesion thickness between the two groups (P < 0.05). The completed resection rate of B group were significant higher than A group (P < 0.05). There was no significant difference in the negative rate of vertical margin between the two groups (P < 0.05). The complication incidence of B group were significant higher than A group (P < 0.05). Conclusion EMR and ESD of endoscopic surgery in the treatment of patients with neuroendocrine tumors of digestive tract possess the same clinical effects; EMR application can efficiently shorten the operation time and decrease the economic burden and ESD application maybe helpful to reduce the complication risk.

OBJECTIVETo confirm the diagnosis of multiple carboxylase deficiency (MCD) on the gene level and explore the mutations in Chinese children with MCD.

METHODSBiotinidase (BT) and holocarboxylase synthetase (HLCS) genes were analyzed by PCR and direct sequencing for the 4 BT deficiency patients and 8 HLCS deficiency patients, respectively. The identified mutations in the parents of the patients and 50 normal controls were screened by PCR-restriction fragment length polymorphism and direct DNA sequencing.

RESULTSTotal detection rate of gene mutation is 100% in the 12 children with MCD. Six mutations were detected in the 4 children with BT deficiency, they were c. 98-104del7ins3, c. 1369G>A (V457M), c. 1157G>A(W386X), c. 1284C>A(Y428X), c. 1384delA and c. 1493_1494insT. The last four were novel mutations. Four mutations were found in the 8 children with HLCS deficiency. They were c. 126G>T (E42D), c. 1994G>C (R665P), c. 1088T>A (V363D) and c. 1522C>T (R508W). The last two were hot-spot mutations [75%(12/16)], and c. 1994G>C (R665P) was a novel mutation.

CONCLUSIONThis study confirmed the diagnosis of 12 patients with MCD on the gene level. Six mutations were found in the BT gene and 4 in the HLCS gene, including 5 novel mutations. Two mutations of the HLCS gene are probably hot-spot mutations in Chinese children with HLCS deficiency.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; Biotinidase ; genetics ; Biotinidase Deficiency ; Carbon-Nitrogen Ligases ; deficiency ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Molecular Sequence Data ; Multiple Carboxylase Deficiency ; genetics ; metabolism ; Mutation

OBJECTIVETo investigate the use of a 32P application device (AD) in the treatment of condyloma acuminatum (CA) in the rectum, and to compare its clinical effect with that of the microwave therapy.

METHODSThis study included 107 cases of CA in the rectum, 99 males and 8 females, aged 21-58 (33.6 +/- 9.4) years. Forty-six of the patients (the AD group) were treated with a self-made 32P application device, which, as a tube-shaped carrier of radionuclide 32P colloid, was fixed in the rectum at the diseased part for medication at 4.9-8.2 Gy for 3-5 hours once and 1-2 times a week. The other 61 (the microwave group) were treated by microwave burning under local anesthesia. Both groups of patients were followed up for over 3 months for comparison of the therapeutic results and observation of the stability and reliability of the 32P application device.

RESULTSThe rates of cure, reoccurrence and adverse reaction were 84, 8%, 13.0% and 8.7% in the AD group, compared with 40.3%, 55.7% and 75.4% in the microwave group, with statistically significant differences between the two groups (P < 0.01).

CONCLUSIONThe 32P application device, with its advantages of low cost, easy operation, good effect, high safety and reliability, low recurrence, fewer adverse events and good acceptability, is highly valuable for the treatment of CA in the rectum.

Adult ; Condylomata Acuminata ; therapy ; Drug Delivery Systems ; instrumentation ; Female ; Humans ; Male ; Microwaves ; therapeutic use ; Middle Aged ; Phosphorus Radioisotopes ; administration & dosage ; therapeutic use ; Rectal Diseases ; therapy ; virology ; Young Adult

OBJECTIVETo study the incidence of various enzyme deficiency in tetrahydrobiopterin (BH4) metabolism and the related gene mutation among the patients with motor disturbance and mental retardation.

METHODSOne hundred patients with unknown motor disturbance and mental retardation were referred to this study. All patients were performed by phenylalanine (Phe) and BH4 loading test, urinary pterin analysis and dihydropteridine reductase (DHPR) activity. Some patients received the dopa treatment for diagnosis of dopa-responsive dystonia (DRD). The analysis of GTP cyclohydrolase 1 gene (GCH1) mutation for DRD patients and the analysis of 6-pyruvoyl tetrahydropterin synthase (PTS) gene mutations for PTS deficient patients were done under the consent from their parents.

RESULTSSeventy of 100 patients had normal basic blood Phe levels, six (6%) patients were diagnosed as DRD. Thirty patients had hyperphenylalaninemia (HPA), eight (8%) were diagnosed as PTS deficiency and 22(22%) were diagnosed as phenylalanine hydroxylase (PAH) deficiency. All patients had normal DHPR activity. The mutation IVS5+3insT of GCH1 was found in 2 patients with DRD. Seven kinds of PTS mutations were found in 8 patients with PTS deficiency, and 75% of the mutations were 259C-->T,286G-->A and 155A-->G.

CONCLUSIONSome patients with unknown motor disturbance and mental retardation may suffer from BH4 metabolism related diseases. Theses patients are necessary to be screened for such kind of diseases in order to confirm the diagnosis.

Adolescent ; Biopterin ; analogs & derivatives ; metabolism ; Child ; Child, Preschool ; Dihydropteridine Reductase ; genetics ; metabolism ; Dystonia ; genetics ; metabolism ; Female ; GTP Cyclohydrolase ; genetics ; metabolism ; Humans ; Infant ; Intellectual Disability ; genetics ; metabolism ; Male ; Mutation ; Phenylalanine Hydroxylase ; genetics ; metabolism ; Phosphorus-Oxygen Lyases ; genetics ; metabolism

AIMTo observe mRNA expression of muscarinic acetylcholine receptors in spinal cord and brainstem in morphine dependent or withdrawal rats.

METHODSThe mRNA expression level of m1, m2, m3, m4 and m5 were determined by RT-PCR, the beta-actin mRNA expression was used as internal control.

RESULTSThe mRNA level of m1, m2, m3, m4 and m5 in spinal cord and m1 and m2 in brainstem were increased significantly during morphine dependence, and the levels of m1, m2, m3 and m4 in spinal cord and m1 in brainstem were decreased 1 h after the injection of naloxone (4 mg.kg-1, i.p.) in morphine dependent rats. Either scopolamine (0.5 mg.kg-1) or pirenzepine (10 mg.kg-1) was shown to significantly decrease the morphine withdrawal symptoms in rats. The levels of m1, m2, m3 and m5 in spinal cord were increased by pretreatment with pirenzepine and the levels of m2, m3 and m4 in spinal cord were increased by pretreatment with scopolamine.

CONCLUSIONThe adaptive expression of muscarinic receptors at spinal and supraspinal levels play important role in mediating morphine dependence and withdrawal in rats.

Animals ; Brain Stem ; drug effects ; metabolism ; Gene Expression ; drug effects ; Male ; Morphine ; toxicity ; Morphine Dependence ; metabolism ; RNA, Messenger ; biosynthesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Receptors, Muscarinic ; biosynthesis ; classification ; genetics ; Spinal Cord ; drug effects ; metabolism ; Substance Withdrawal Syndrome ; metabolism

OBJECTIVETo observe the ECG and electrophysiological characteristic of patients with idiopathic ventricular tachycardia (VT) and premature ventricular contraction (PVC) originating from left (LVOT) and right (RVOT) ventricular outflow tracts and assess the clinical effect of radio frequency catheter ablation (RFCA) on these patients.

METHODSRFCA was performed in 58 patients (10 with VT and 48 with PVC, 5 patients with VT from RVOT under the guidance of non-contact mapping system Ensite3000). VT or PVC originated from LVOT in 15 patients (12 out of 15 from left sinus of Valsalva) and RVOT in 43 patients.

RESULTS(1) R wave in II, III, aVF leads was the common characteristics of VT or PVC originated from LVOT and RVOT and difference in wave duration index and R/S-wave amplitude ratio in V(1) or V(2) could be used to define VT and PVC originated from LVOT or RVOT. (2) Ablation was successful in 55 out of 58 patients (9 patients with the 2nd ablation, evaluated as arrhythmia-free at 3 months post ablation without medication) and failed in 3 patients. One patient developed pericardial tamponade during ablation and recovered without complication after related treatments.

CONCLUSIONSRFCA is an effective, safe and curative therapy for VT or PVC originated from LVOT and RVOT. Non-contact mapping system (Ensite3000) is a safe and reliable tool to guide mapping and ablation in patients with complex VT and unstable hemodynamics.

Adolescent ; Adult ; Aged ; Catheter Ablation ; Female ; Humans ; Male ; Middle Aged ; Tachycardia, Ventricular ; etiology ; therapy ; Ventricular Outflow Obstruction ; complications ; Ventricular Premature Complexes ; etiology ; therapy ; Young Adult

OBJECTIVETo investigate the incidence of hyperphenylalaninemia (HPA) caused by different etiologic factors in China and the relationship between the phenylalanine and mental development of patients with HPAs who were diagnosed by neonatal screening and early treated.

METHODSTwo hundred and twenty-three patients with HPA detected by neonatal screening programs were refered to us at the age of (41 +/- 27) days after birth. The differential diagnosis was performed by BH(4) (20 mg/kg) loading test, urinary pterin analysis and dihydropteridine reductase (DHPR) activity determination respectively. The control of phenylalanine (Phe) metabolism, growth and mental development were evaluated in all treated patients. Related gene mutation analysis was performed in some patients

RESULTSOne hundred and twenty-nine of 223 patients (57.8%) were diagnosed as phenylalanine hydroxylase deficiency (PAHD), 64 patients (28.7%) as BH(4) responsive PAHD, 30 patients (13.5%) as 6-pyruvoyl tetrahydropterin synthase deficiency (PTSD). One hundred and forty-nine patients were followed at age of 4 m - 2 y in our clinic. The 136 of 149 patients were treated according to different etiology at the age of 1.6 m (0.5 - 3.5 m) after birth. Thirteen patients were followed up without the need for treatment. All patients had normal growth development. One hundred and eight (79.4%) of 136 treated patients had normal mental development. The negative correlation (r = -0.439, P < 0.01) between IQ and average Phe levels were observed in 58 patients. Twenty-eight patients were able to go to primary school or even university. Nine kinds of PTS gene mutations were found in 9 cases with PTSD, among which 286G-->A and 259C-->T were most commonly seen, accounting for 45%. Seven kinds of PAH gene mutations were found in 13 cases with BH(4) responsive PAHD with the R241C (43.8%) mutation being the most frequent one.

CONCLUSIONThe differential diagnosis should be quickly made in all HPA patients detected by neonatal screening. Near 80% patients early treated had normal mental development. The good control of blood Phe level is a key factor for mental development.

China ; epidemiology ; Female ; Follow-Up Studies ; Humans ; Incidence ; Infant ; Infant, Newborn ; Male ; Neonatal Screening ; methods ; Phenylalanine ; blood ; Phenylketonurias ; diagnosis ; epidemiology ; prevention & control ; Time Factors

The antisense approach and RT-PCR were used to study the effects of muscarinic receptors on the scores of morphine-withdrawal syndrome and the expression of NMDA receptor subtypes (NR(1A) and NR(2A)) mRNA in rat spinal cord and brainstem. The concentrations of glutamate in periaqueductal grey (PAG) of morphine-withdrawal rats were determined by capillary electrophoresis with laser-induced fluorescence detection. The data showed that the NR(1A) and NR(2A) mRNA levels were increased significantly in the spinal cord and brainstem 1 h after the injection of naloxone (4 mg/kg, i.p.) in morphine-dependent rats. Moreover, in morphine-dependent rats pretreated (i.p.) with scopolamine (0.5 mg/kg), or pirenzepine (10 mg/kg), MK801 (0.125 mg/kg), L-N-nitroarginine methylester (10 mg/kg) 30 min before naloxone injection, the NR(1A) and NR(2A) mRNA levels were significantly lower than those of 1 h morphine-withdrawal rats. Intrathecal injection of NR(1A) or M(2) receptor antisense oligonucleotides (A-oligo, 4 microg/per rat) 24 h prior to naloxone challenge could block the morphine withdrawal symptoms including wet dog shaking, irritability, salivation, diarrhea, chewing and weight loss. Meanwhile, in morphine-dependent rats the NR(1A) mRNA levels in the spinal cord and brainstem were down-regulated by intrathecal injection of M(2) receptor A-oligo. The glutamate concentrations in PAG microdialysis were increased to a maximal level 15 min after naloxone injection. The glutamate response was inhibited by pretreatment with M(2) receptor A-oligo but not by M(1) A-oligo. The results suggest that the expression of NMDA receptors and the release of glutamate in brainstem are involved in the processes of morphine withdrawal and that the NMDA receptor expression is possibly regulated by the muscarinic receptors during morphine withdrawal.
Animals ; Brain Stem ; metabolism ; Glutamic Acid ; metabolism ; Male ; Morphine ; adverse effects ; Periaqueductal Gray ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Muscarinic ; physiology ; Receptors, N-Methyl-D-Aspartate ; biosynthesis ; genetics ; Spinal Cord ; metabolism ; Substance Withdrawal Syndrome ; genetics ; metabolism