"CapFinder" technology,which can be used to clone the full length of 5′ UTR sequence of mRNA,was described.This technology used the terminal transferase activity of certain MMLV RT variants that added 3-5 residues(predominantly dC) to the 3′end of the first-strand cDNA exhibited when MMLV RT reached the 5′cap structure of mRNA.In the reverse reaction system containing GGG oligo,the terminal transferase activity was harnessed by the GGG oligo whose terminal stretch of dG residues can anneal to the dC-rich cDNA tail and serve as an extended template for RT.After RT switch templates from the mRNA template to the GGG oligo,a complete cDNA copy of the original RNA was synthesized with the additional GGG oligo sequences at the end.5′UTR of mRNA can be amplified with GGG oligo as forward primer and a gene-specific reverse primer.5′UTR of Bt toxin receptor E-Cadherin gene in midgut of cotton bollworm was cloned.

Noise, Occupational ; prevention & control ; Occupational Health Services ; standards ; Threshold Limit Values

Adolescent ; Adult ; Dust ; Female ; Humans ; Male ; Mental Health ; statistics & numerical data ; Middle Aged ; Occupational Exposure ; Steel ; Surveys and Questionnaires

Objective To observe the behavior changes and synaptic reconstruction of brain and the impact of gastrodin in rats with epilepysy induced by pentylenetetrazole.Methods Fifty Wistar rats in growth(50-70 g) were randomly divided into 5 groups:control group,Pentylenetetrazole (PTZ) group,gastrodin high dose group,gastrodin low dose group and sodium valproate group,each group had 10 rats.According to Racine classification,the behavior changes of rats and the frequency were recorded.Conventional method was adopt to perfuse cordis, fix and extract the brain,and the immunohistochemistry was used to detect synaptophysin(P38) expression in the temporal lobe and hippocampus of the brain after 4 weeks.Results 1.Four weeks after pentylenetetrazole ignited the growth period rats,the attack-level of each experimental group through comparison with each other,there was statistically significant difference(?2=35.83 P0.05).Conclusions There are mossy fiber sprouting and the formation of synaptic reconstruction in temporal lobe and hippocampus of the growth period rats repea-tedly ignited by pentylenetetrazole.Gastrodin may play a role in the formation of antiepileptic obviously,and through decreasing the expression of P38,which can inhibit the formation of synaptic reconstruction,as control seizures indirectly.

Objective To explore the clinical significance of the blood lactic dehydrogenase(LDH), ?_2-microglobulin(?_2-MG),D-dimer measuring in the diagnosis and treatment of Non-Hodgkin lymphoma. Methods In 40 cases with NHL,LDH was measured by L-P continuous monitoring method,?_2-MG was measured by luminescent immunoassay,D-dimer was measured by immunoturbidimettic assay.Results The levels of the blood LDH,?_2-MG and D-dimer in patients with NHL were higher than those of in the controls(P 0.05).Con- clusion The levels of blood LDH,?_2-MG,D-dimer can be taken as an auxiliary clinical index to diagnose, classify the phase,evaluate the effectiveness of treatment and prognosis in the NHL patients,and have impor- tant clinical significance.

Objective To discuss the strategy and intervention measures of the iodine deficiency disorders (IDD)control in coastal salt.producing areas so as to shoot the problem of non-iodized salt causing IDD.Methods Accordinng to different areas,periods and crowds,eomplicatd measures and strategies were taken such as supply of iodized salt to peopie in special need while universalizaion of iodized salt,health promotion,private salt factory censu8 and close.iodized salt quality monitoring and promotion of technology of iodized salt producion in Xiamen, where the probiem of non-iodized salt Was serious since 1995.Results Iodized salt manufactured Was qualified in a increased rate from 89.50% in 1995 to 96.17% in 1997,stablized at 99.00%since 2000.Qualified iodized salt sold in the shops waft increased from 87.33%in 1996 to 96.33%in 1998.Popularization covered by iodized salt in urban areas increaased from 0.92%in 1 995 to 100.00%in 2000,and it Was increased from 0 to 99.00%in suburban area8 and increased from 0 to 89.00%in rural areas.Since 2001 iodized salt covered up 93.00%of the people.The rate of child goitre in urban,suburban and rural areas respectively Wag 16.44%(228/1387), 20.57%(266/1293) and 24.93%(651/2611).Moreover,beginning from 1996,it reduced tO below of 5.00%respectively in 1999,2001 and 2005.The median of urinary iodine of children in urban,suburban and rural areas respectively was 137.50, 102.12,94.66 μg/L in 1995,since 1997 it reached 100.00μg/L and kept at 120.00μg/L In 2007 the median of urinarv iedine of children respectively was 271.10,240.40,198.10μg/L in urban,suburban and rural areas.The pereentage of awareness of IDD knowledge was 74.00%(444/600)in students in 1997 and reached 95.00% since 2000.Conclusion The paRern of eliminating iodine deficiency dis ease in Xiamen has successful established,which works efficiently and sets an example for iodized salt supplement in non-iodized salt areas and continually eliminating the iodine deficiency disease.

AIMTo determine the role of area postrema (AP) of rabbit in the regulation of cardiovascular function.

METHODSThe rabbits were anesthetized with intravenous injection of 10% urethane and 1% chloralose, and were artificially ventilated. The changes of mean arterial pressure (MAP) and heart rate (HR) were observed when AP was electrically stimulated with different frequency (10 Hz -80 Hz) and after chemical lesion of CVLM or RVLM, respectively.

RESULTSElectrical stimulation of AP with low frequency (10 Hz, 20 Hz) decreased MAP and HR. Stimulation with high frequency(60 Hz, 80 Hz) increased MAP but decreased HR. The changes in MAP and HR were significantly lower (P < 0.01) after CVLM was destroyed when electrical stimulation of AP with 20 Hz, and both changes of MAP and HR were disappeared (P < 0.01) after RVLM was destroyed when electrical stimulation with 20 and 80 Hz.

CONCLUSIONElectrical stimulation of AP with low frequency decreases MAP and HR, stimulation with high frequency induces an increase in MAP and decreases in HR. The former is probably related to excitation of CVLM, the cardiovascular effects induced by different frequency of electrical stimulation are all resulted from the activation of RVLM.

Animals ; Area Postrema ; physiology ; radiation effects ; Blood Pressure ; radiation effects ; Electric Stimulation ; Heart Rate ; radiation effects ; Male ; Medulla Oblongata ; physiology ; radiation effects ; Rabbits

AIMTo investigate the cardiovascular response caused by intracerebroventricular (ICV) microinjection of interleukin-2 (IL-2) and explore the underlying mechanism.

METHODSMale Sprague-Dawley rats were anesthetized with intraperitoneal urethane( 1.2 g/ kg). The changes of mean arterial blood pressure (MAP) and heart rate (HR) were observed during ICV microinjection of IL-2 with or without pretreatment of naloxone or atropine or phentolamine.

RESULTSThere were no significant effects on cardiovascular response after ICV injection of IL-2 at 500 IU/3 microl and 1 000 IU/3 microl, but IL-2 at 1 500 IU/3 microl could elevate MAP and HR. The responses of MAP and HR reached their maximum levels at 10 min (MAP: 10 +/- 1.8 mmHg, HR: 25 +/- 2 b/min, P < 0.05) after the injection and lasted 15 or 10 minutes respectively. Pretreatment with naloxone (10 microg/10 microl) or atropine (1.5 microg/10 microl) could block the cardiovascular response of ICV injection of IL-2. Pretreatment with phentolamine (10 microg/10 microl) failed to block the cardiovascular responses by IL-2.

CONCLUSIONICV microinjection of interleukin-2 (IL-2) can elevate the MAP and HR, which may be mediated by central opioid and cholinergic system. The alpha-adrenergic system may be not involved in the cardiovascular response of IL-2.

Animals ; Blood Pressure ; drug effects ; Heart Rate ; drug effects ; Injections, Intraventricular ; Interleukin-2 ; administration & dosage ; pharmacology ; Male ; Microinjections ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effects of Bufei Jianpi Recipe (BJR) on the diaphragmatic neural discharge and the diaphragmatic muscle function in rats with chronic obstructive pulmonary disease (COPD).

METHODSRats were randomly divided into the normal control group, the model group, the high dose BJR group (9.68 g/kg x d(-1)), the medium dose BJR group (4.84 g/kg x d(-1)), the low dose BJR group (2.42 g/kg x d(-1)), and the aminophylline group (2.3 mg/kg x d(-1)). The stable phase COPD rat model was prepared using repeated smoke inhalations and bacterial infections. The high, medium, and low dose BJR and aminophylline was respectively administered to rats from the ninth week to the twentieth week. The sampling was taken. The lung function, diaphragmatic neural discharge time (Td), and diaphragmatic neural discharge interval (Tdi), diaphragmatic neural discharge range (Rd), diaphragmatic neural discharge area (Ad), expiratory time (Tex), inspiratory time (Tin), respiratory rate (RR), respiratory excursion (RE), respiratory area (RA), and diaphragmatic muscular tension and endurance were detected.

RESULTSCompared with the normal control group, the tidal volume (TV), peak expiratory flow (PEF), and 50% tidal volume expiratory flow (EF50) significantly decreased in the model group (P < 0.01). Td, Tdi, Tex, and Tin were significantly prolonged (P < 0. 05, P < 0.01). Ad, Rd, RR, RE, RA, diaphragmatic muscular tension and endurance significantly decreased (P < 0.05, P < 0.01). The ratio of type I and IIA diaphragmatic fibers significantly increased and type IIB significantly decreased (P < 0.01). The activity of ATP decreased and the activity of SDH increased (P < 0.01). The aforesaid indices were improved to different degrees in BJR groups, especially in the high dose BJR group and the medium dose BJR group (P < 0.05, P < 0.01).

CONCLUSIONSBJR could significantly improve the diaphragmatic neural discharge and the diaphragmatic muscle function. Its efficacy was better than that of aminophylline.

Aminophylline ; pharmacology ; Animals ; Diaphragm ; drug effects ; physiopathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Phrenic Nerve ; drug effects ; physiopathology ; Phytotherapy ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; physiopathology ; Rats

Background Our previous study demonstrated that curcumin can induce the apoptosis of retinal pigment epithelial (RPE) cells and herein inhibit the proliferation of RPE cells,and it is proved that the intravitreous injection of 0.1mg curcumin has less adverse effect to ocular tissue, inferring a good applicative prospect in clinic. Objective The goal of this experiment was to evaluate the effectiveness of curcumin on the prevention and treatment of experimental proliferative vitreoretinopathy (PVR). Methods PVR models were induced by injection of 0.1ml RPE cells (containing 2×106 cells) into vitreous cavity in 40 eyes of 20 healthy and mature New Zealand albino rabbits.0. 1ml curcumin(0. 1 mg) was then injected into lateral eye of each model rabbit immediately following the injection of RPE cells,and the equal volume of normal saline solution containing 0. 5‰ DMSO was injected into the fellow eye of each model rabbit as controls. On 1,3,7,14,21 and 28 days after injection, the changes of cornea, aqueous humor, lens, vitreous and fundus were examined and recorded by slit lamp biomicroscope, indirect ophthalmoscope,fundus color camera and B-type ultrasonograph to evaluate the inflammatory response. The incidence rate of retinal detachment was calculated and compared between curcumin group and control group. Results The inflammatory reaction in anterior chamber and misty opacity in vitreous were found from 1 day through 3 days after injection, but no obvious proliferative strap and retinal detachment in all of the experimental eyes. On the 7th day after injection, inflammatory reaction was extinct in the anterior chamber of rabbit eyes, and proliferative strap occurred in 14 eyes(75% ) in the control group but only 2 eyes (10% ) in curcumin group,showing significant difference between these two groups (P<0. 01). No retinal detachment was seen in both the two groups. On 14,21 and 28 days after injection, the incidence rate of retinal detachment was 55% ,80% ,95% respectively in control group and that of curcumin group was 10% ,15% ,15% respectively,presenting considerably differences between two groups (P<0. 01, P<0. 01 ,P<0. 01 ). Conclusion Injection of curcumin into vitreous cavity can effectively inhibit the occurrence and development of PVR in rabbit.