Objective To investigate the DNA methylation feature and DNA methylation regulation to its transcription and expression of O6-methylguanine-DNA methyltransferase gene (MGMT) in NaAsO2-treated HaCaT cells. Methods HaCaT cells were treated 72 hours at intervals and repeatedly by 3.13, 6.25,12.50, and 25.00 μmol/L NaAsO2, MGMT gene promoter region was amplified in the transcription initiation site - 329 - + 93 region by bisulfate-sequencing polymerase chain reaction (BSP), the mRNA transcription and the protein expression of MGMT was detected by real-time quantitative PCR and Western blotting. NaAsO2-untreated HaCaT cell was set as a blank control, and human epidermal squamous carcinoma cell strain A431 was set as a positive control. Results Among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, the positive rates of the DNA methylation of promoter region in MGMT gene were 0.63%(l/160), 6.25% (10/160), 10.63%( 17/160) and 18.75% (30/160), respectively, and methylated CpG sites were mainly located in - 249--146 region relative to transcription start site. There was no DNA methylation in the blank control. There were significant differences between the blank control and the NaAsO2-treated cells (x2 = 76.687, P< 0.05). Average levels of MGMT mRNA were 1.518 31 ± 0.180 54, 1.425 22 ± 0.180 39, 1.014 54 ± 0.096 79 and 0.887 72 ± 0.020 00, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells(1.198 29 ± 0.159 97), there were significant differences(F = 37.359, P < 0.05). Average levels of MGMT protein were 1.174 47 ± 0.064 75, 0.848 83 ± 0.057 01, 0.471 63 ± 0.023 34 and 0.240 34 ± 0.014 43, respectively among the groups of HaCaT cells treated with 3.13, 6.25, 12.50 and 25.00 μmol/L NaAsO2, compared with the blank control cells (1.066 19 ± 0.061 24), there were significant differences(F = 20.687, P < 0.05). Conclusions Arsenic can cause CpC island hypermethylation in the promoter region of MGMT gene, which results in inhibited MGMT mRNA transcription and protein expression. It might be one of the important mechanisms of arsenic-induced skin lesion.

Objective To evaluate denervation changes [fibrillation potentials (fib) and positive sharp waves (psw)] in thoracic paraspinal muscles as diagnostic aids in amyotrophic lateral sclerosis (ALS). Methods The fib and psw of thoracic paraspinal muscles,the motor unit potential of sternocleidomastoid muscle were detected in 50 patients who had been identified as having ALS.Age and sex-matched,30 normal subjects and 30 patients who suffered radicular disease in cervical or lumbosacral region were also detected in thoracic paraspinal muscles for finding fib and psw serving as controls. Results In ALS group,a vast of fib and psw were found in thoracic paraspinal muscles in 41 of 50 ALS patients (82%),they were not found with sternocleidomastoid muscle,but the motor unit potential of this muscle was shown having long-duration and high-amplitude in 48 of 50 ALS patients (96%),which were coincided with the chronic neurogenic changes.6 of 50 patients had denervation changes of tongue muscles,3 of them found fib and psw.2 of 30 patients who suffered from radicular diseases were found with a few of fib and psw changes in thoracic paraspinal muscles.None of these symptoms were found in normal subjects.Conclusion A vast of fib and psw in thoracic paraspinal muscles should represent a sensitive tool for diagnosis of ALS.

Objective To evaluate the incidence of asymptomatic carotid stenosis (ACS) in patients with peripheral arterial occlusive disease (PAOD) who was admitted for revascularization.Methods 229 PAOD patients were evaluated by carotid ultrasound preoperatively and divided into severe ACS group (ACS ≥ 70％) and non severe ACS group (ACS ＜ 70％).The clinical characteristics and associated risk factors of the two groups were analyzed.Results The incidence of severe ACS was 19.7％,age,blood pressure,total cholesterol and fibrinogen levels in severe ACS group were significantly higher than those in non-severe ACS group (P ＜ 0.05),the more severe the symptoms of PAOD are,the higher is the incidence of severe ACS.Age,hypertension and hypercholesterolemia were all predictors of severe ACS.Conclusions The severity of PAOD is positively correlated with the incidence of severe ACS.Age,hypertension and hypercholesterolemia are independent risk factors for severe ACS.

Objective To compare the postoperative follow-up indexes of femoral popliteal artery bypass grafts and evaluate the sensitivity of vessel function test methods for the early detection of grafts restenosis.Methods 84 cases were followed-up after femoral popliteal graft bypass,using ankle brachial index (ABI),ABI decrease amplitude,and Duplex ultrasound to detect peak systolic velocity (PSV) and peak systolic velocity ratio (PSVR).All patients were divided into mild stenosis group (0-49％),moderate stenosis group (50％-74％) and severe stenosis group (75％-99％).Results ABI was 0.88 ±0.12,0.81 ± 0.10 and 0.52 ± 0.13;ABI decrease amplitude was 0.13 ± 0.06,0.12 ± 0.06 and 0.36±0.06;PSV(m/s) was 0.89 ±0.23,0.91 ±0.31 and 1.70 ±0.85,respectively.There was remarkable difference for ABI,ABI decrease amplitude and PSV between severe stenosis and mild to moderate stenosis group (P ＜0.05).PSVR was 1.00 ±0.08,2.14 ±0.31 and 3.58 ±0.31 respectively,in the three groups (P ＜ 0.05).Conclusion PSVR was more sensitive to detect grafts restenosis early than ABI,ABI decrease amplitude and PSV.

Objective To explore the clinical value of the duplex ultrasonography (duplex US) for evaluating the re-stenosis after peripheral arterial bypass grafting. Methods Eighty prosthetic grafts of sixty-three patients with femoral-pop-liteal arterial bypass grafting were follow-up regularly by duplex US. They were divided into non significant stenosis group (n=56), the significant stenosis group (n=15) and occlusion group (n=9) according to the tube diameter and arterial blood flow-ing parameters, which changed postoperatively. The diagnostic results were compared and analyzed between duplex US and digital subtraction angiography (DSA). The peak flow velocity of middle grafts (MG) to 40 cm/s was defined to evaluate risk of graft occlusion. Results The diagnostic coincidence rate of duplex US and DSA for grafts stenosis classification was 90%. The diagnostic sensitivity of duplex US to grafts stenosis was 91.7%, and the specificity was 92.9%. The positive pre-dictive value was 84.6%for grafts stenosis, and the negative predictive value was 96.3%, the false positive rate was 16.7%, and the false negative rate was 8.3%. The grafts occlusion rate was higher in MG<40 cm/s group than that of MG≥40 cm/s group. Conclusion There was a good consistency with Duplex US and DSA for the diagnosis of peripheral artery bypass graft restenosis. Duplex US showed characteristics of non-invasive, simple and easily accepted by patients.

China ; Gastroesophageal Reflux ; therapy ; Humans ; Integrative Medicine ; Medicine, Chinese Traditional ; methods

OBJECTIVETo investigate the effects of epigallocatechin gallate (EGCG)against exercise-induced fatigue in mice.

METHODSTotal 120 mice were randomly divided into three groups and tested separately. For each test, there were 30 mice subdivided into high dose (50 mg/kg . d EGCG) and low dose (10 mg/kg . d EGCG) groups as well as saline control group(1 ml/kg . d) with 10 in each. Burden swimming, running wheel endurance, stick climbing and hypoxia tolerance exercise were used to establish fatigue mice training model in three groups. And intraperitoneal injection with different doses of EGCG per day for consecutively 28 days and the mice in the control group were treated with normal saline. After the last each test, the blood lactic acid (BLA), blood urea nitrogen (BUN), blood lactate dehydrogenase (LDH), muscle glycogen (MG) and liver glycogen (LG) of each group of mice were determined.

RESULTSEGCG treatment groups(B and C)revealed a prolonged the mice survival time of burden swimming test, hypoxia tolerance, running wheel time and the ability of stick climbing(P < 0.05 or P <0.01), and increased LDH activity and MG and LG contents, reduced contents of BLA and BUN. High dose group had an obviously increase effect than lower dose group(P <0.05).

CONCLUSIONEGCG has significant effects against exercise-induced fatigue in mice.

Animals ; Blood Urea Nitrogen ; Catechin ; analogs & derivatives ; pharmacology ; Exercise Tolerance ; Fatigue ; drug therapy ; Glycogen ; metabolism ; L-Lactate Dehydrogenase ; blood ; Mice ; Physical Conditioning, Animal

Animals ; Catechin ; analogs & derivatives ; pharmacology ; Disease Models, Animal ; Fatigue ; metabolism ; physiopathology ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism

AIM: To study the expression of MDM2 and muta nt-type P53 proteins in primary hepatocellular carcinoma(HCC). METHODS: Using immunohistochemical staining method(SP),the expression of mutant-t ype P53 and MDM2 proteins was examined in 55 cases of HCC,23 cases of correspond ing paracancerous tissues and 10 cases of normal hepatic tissues. RESULT S: The frequencies of MDM2 and P53 positive expression in HCC were 17/55 (30 9%) and 23/55 (41 8%), respectively. There was both positive expression of MDM2 and mutant-type P53 in 11 cases (20%) with HCC. Expression of MDM2 showed a significantly positive correlation with expression of mutant-type P53 (r=0 310,P

Objective To investigate the transcription and expression of DNA methyltransferase 1 (DNMT1) mRNA in endemic arsenism patients by burning coal usage,to probe its effects on the development and carcinogenesis of arsenism. Methods In 2008,68 arsenism patients(including 24 mild cases,28 moderate cases and 16 severe cases) were selected in the areas with endemic arsenism according to Standarding of Diagnosis for Endemic Arsenism from Xingren county,Guizhou province. Among the subjects,40 cases were diagnosed by pathological methods,and they were divided into general pathological changes(20),precancerous(14) and cancerous group(6). Tweleve kilometer away from the endemic arsenism area,23 controls were selected in Daguoduo village (non-arsenism exposure). Under the principle of informed consent,blood samples were collected from individuals. The mRNA expression of DNMTI was detected by real-time quantitative reverse transcription polymerase chain reaction(FQ-PCR). At the same time,skin tissue samples were collected from the voluntary surgical treatment patients with endemic arsenism (total 61 cases,including 34 general pathological changes cases,21 precancerous cases and 6 cancerous cases) and from the control(15 cases). DNMT1 protein was detected by immunohistochemical method.Results Average level of DNMT1 mRNA were 0.221 83±0.595 09,0.246 11±0.509 79 and 0.389 27±0.411 33 respectively among mild,moderate and severe arsenism group. DNMT1 mRNA level of mild and moderate group were obviously lower than the control group(0.695 95±0.463 98,all P < 0.01). The mRNA average level of DNMT1 were 0.320 64±0.547 46,0.313 09±0.529 13 and 0.159 07±0.342 56 individually among general pathological changes,precancerous and cancerous group,which were obviously lower than the control group(0.695 95±0.463 98,all P < 0.05). The expression rates of DNMT1 protein in skin were 88.24%(30/34),100%(21/21) and 100% (6/6) among general pathological changes,precancerous and cancerous group were higher than the control group [0(0/15),all P < 0.01],and the extent of expression gradually increased with the aggravation of skin damage(r,= 0.740,P < 0.01). Conclusions DNMT1 participated in the development of the arsenism. High expression of its protein was an early event during the process of the arsenism. DNMT1 may be the new target markers for early diagnosis and treatment of arsenism.