0.05),but the average levels of DNMT1 mRNA of mild and moderate groups were significantly lower than that of the control group(P

Objective To investigate the effects of Mailuoning on oxidative stress and ischemic brain injury. Methods A total of 126 healthy male Kunming mice were divided into sham operation (n = 18), normal saline control (n = 54) and Mailuoning (n = 54) groups. A middle cerebral artery occlusion (MCAO) model was induced Two hours after MCAO,Mailuoning injection and equivalent saline were injected via the tail vein in the Mailuoning and normal saline control groups, respectively, and then they were injected every other 24 h.Neurological score was performed, and brain water content, infarct volume, membrane potential,as well as protein oxidative stress metabolites such as 3-nitrotyrosine (3-NT), lipid oxidative stress metabolite 4-hydroxy-2-nonenal (HNE) and nucleic acid oxidative stress metabolite 8-hydroxy-2'-deoxyguanosine (8-OHdG) were detected at 12, 24 and 72 h after MCAO. Results Mailuoning injection could significantly improve the neurological function of cerebral ischemia in mice, decrease brain edema, and reduce infarct volume at different time points after cerebral ischemia Of those, it was most significant at 72 h. Mailuoning injection could reverse the decreased mitochondrial membrane potential in cerebral cortex and internal capsule, and significantly downregulate the increased 3-NT, HNE and 8-OHdG in cerebral cortex, internal capsule and serum after ischemia, of those, the effect of reducing HNE was most significant.Conclusions Mailuoning injection may effectively protect against ischemic brain injury in mice,and its mechanism is associated with inhibiting oxidative stress, particularly anti-lipid oxidation.

Background Leber hereditary optic neuropathy (LHON) is a maternally inherited disease caused by mitochondrial DNA (mtDNA) mutation with the common mutation sites of m.3460 G＞A,m.11778 G＞A and m.14484 T＞C,and other mutation sites are rare.Understanding the mutation type of mtDNA in LHON patients has an important clinical significance.Objective This study was to analyze the clinical features of LHON and detect the mitochondrial mutation.Methods Twelve unrelated Chinese patients who was diagnosed as LHON were included in Peking University People's Hospital from 2010 to 2014.The visual acuity,perimetry,ocular segment,visual evoked potential,fundus were binocularly examined.The peripheral blood of 4 ml was collected from each patient and mtDNA was amplified and sequenced by using PCR.Three common genetic mutation sites for LHON and other mutation sites were determined and analyzed.This study protocol was approved by Ethic Committee of Peking University People's Hospital and complied with Helsinki Declaration.Written informed consent was obtained from each patient prior to any medical examination.Results Of the 12 patients,11 were male and 1 was female.The visual acuity of both eyes reduced simultaneously in 7 patients,and the visual acuity of left eye and the right eye first reduced in 3 patients and 1 patient,respectively.There was no significant correlation in the visual impairment between the left and right eyes (P＞0.05).In the near vision of the patients,J7 was invisible in 18 eyes,and J7 were obtained in 3 eyes,J6 were obtained in 2 eyes and J2 was obtained in 1 eye.In the distant vision of the patients,hand movement was obtained in 1 eye,light perception was obtained in 1 eye,0.01-0.1 were obtained in 18 eyes and 0.12-0.3 were obtained in 2 eyes.The visual field defect of nasal lateral was found in 7 eyes,visual field defect of temporal lateral was found in 3 eyes and the visual field defect of central was found in 8 eyes.mtDNA sequencing revealed that m.3460 G＞A mutation was seen in 3 patients,m.11778 G＞A mutation was seen in 5 patients and m.14484 T＞C mutation was seen in 2 patients.In addition,other 2 mutations were found in 2 patients,which were m.3497 C＞T and m.10663 T＞C mutations at the MT-ND1 and MT-ND4L genes,respectively.Conclusions LHON is more common in male.Visual impairment shows the varying degrees between both eyes of patients and appears to be severe in near vision.Central visual field defect is common in LHON patients.This study detects m.3497 C＞T and m.10663 T＞C mutation in Chinese LHON patients.

OBJECTIVE To investigate DNA hypermethylation of human 8-hydroxyguanine glycosy-lase(hOGG1 )gene and and the level of oxidative stress and DNA oxidative damage relations with arse-nic poisoning.METHODS In ende mic coal-pollution-borne arsenism area,Xinren county,Guizhou Province,according to the diagnostic criteria of ende mic arsenism(WS /T21 1 -2001 ),207 people with ende mic arsenism were selected and divided into four groups(The arsenic exposure group:46 cases, mild arsenism group:46 cases,moderate arsenism group:60 cases and severe arsenism group:55 cases).64 residents were selected as controls in a village about 12 km away fro m the ende mic arsenism area.With the informed consent principle,peripheral blood of all respondents was collected in order to analyze DNA methylation.Methylation-specific poly merase chain reaction were respectively performed to analyze hOGG1 Hypermethylation in arsenism respondents.Che mical methods were performed on the activity of super oxide dis mutase (SOD)and glutathione peroxidase (GSH-Px),while the contents of malondialdehyde (MDA)in the blood of patients were measured,and the contents of 8-hydroxy-2′-deox-yguanine(8-OHdG)urine of patients were measured and analysed.On the basis of methylation status are divided into hOGG1 gene methylation group (34 cases)and hOGG1 gene no methylation group (237cases).Analysis was performd on hOGG1 gene DNA methylation and the relationship between oxi-dative stress and arsenic poisoning.RESULTS The positive rates of hypermethylation of hOGG1 were associated with the degree of arsenic poisonin (co mpared with control group,χ2 =23.916,P <0.05, Co mpared with the Ende mic area normal group,χ2 =12.039,P <0.05 ).Co mparing with negative group,SOD〔(85 ±25)kU·L -1 〕,GSH-Px〔(70 ±26)kU·L -1 〕activity and 8-OHdG 〔(22.5 ±6.8)μg·L -1 〕contents were lower〔(1 18 ±41 )kU·L -1 ,(171 ±56)kU·L -1 ,(28.4 ±6.5)μg·L -1 ,P <0.05)〕in positive group.There was no significant difference between the MDA content(P>0.05).CONCLUSION Coal arsenic exposure can cause hOGG1 gene high methylation and oxidation and anti-oxidation system imbalance,causing DNA oxidative damage,it is one of the reasons to pro mote the develop ment of arsenic poisoning occurred.

Objective To detect the combining capacity of peripheral blood NF-E2-related factor 2 (Nrf2)of arsenic-exposed residents in the coal-contaminated arsenism area in Guizhou with the sequence of downstream antioxidant response element (ARE) as well as antioxidase gene expression,and to provide a basis for in-depth revelation of arsenic oxidative damage mechanism to human body.Methods Jiaole and Changqing villages in coal-burning-borne arsenism areas in Xingren County of Guizhou were selected as the survey spots,and 161 cases of arsenic-exposed residents were selected as the arsenic exposed group on the basis of physical examination.They were divided into non-patient group (21 cases) and patient group (140 cases) according to the Diagnostic Criteria of Endemic Arsenism (WS/T 211-2001),and the patient group was further divided into mild hepatosis group (52 cases),moderately severe hepatosis group (36 cases) and non-apparent hepatosis group (52 cases) according to the Diagnostic Criteria of Occupational Chronic (GBZ 59-2010).Moreover,45 residents from one village neighboring to non-epidemic area were selected as controls.The hemocyte nucleoprotein was extracted from peripheral blood in the sampling subjects.The combining capacity of peripheral blood Nrf2-ARE was tested by electrophoretic mobility shift assay (EMSA),and the relative expression quantity of Cu/Zn superoxide dismutase (Cu/Zn-SOD) and glutathione peroxidase 1 (GSH-Pxl) mRNA was tested with real-time fluorescence quantification PCR (qPCR).Results The testing results of Nrf2-ARE combining capacity showed that the difference of Nrf2-ARE combining capacity between groups was statistically significant (F =116.033,P ＜ 0.05).Compared with the control group (3.14 ± 1.34),the Nrf2-ARE combining capacity was higher in the non-apparent hepatosis group (5.17 ± 2.06),mild hepatosis group (13.13 ± 4.84) and moderately severe hepatosis group (32.35 ± 14.76,all P ＜ 0.05);compared with the non-patient group (5.15 ± 3.23) and non-apparent hepatosis group,the Nrf2-ARE combining capacity of mild hepatosis group and moderately severe hepatosis group was higher (all P ＜ 0.05);compared with mild hepatosis group,the Nrf2-ARE combining capacity of moderately severe hepatosis group was higher (P ＜ 0.05).The results of Cu/Zn-SOD and GSH-Pxl mRNA expression showed the relative expression quantities of Cu/Zn-SOD [Median (M):1.127 8,1.257 8,1.632 0] and GSH-Pxl (M:1.334 5,1.940 9,2.062 6) mRNA of non-apparent hepatosis,mild hepatosis,moderately severe hepatosis groups were higher than those of the control groups (M:0.961 8,0.884 3),respectively,and their differences were statistically significant (x2 =13.065,19.934,all P ＜ 0.05).The relative expression quantities of GSH-Pxl mRNA of mild hepatosis group and moderately severe hepatosis group were higher than that of the non-patient group (M:1.248 4),and their differences were statistically significant (all P ＜ 0.05).The correlation analysis indicated the Nrf2-ARE combining capacity was positively related to the Cu/Zn-SOD and GSH-Px1 mRNA expression (r =0.271,0.292,all P ＜ 0.01).Conclusion The increase of Nrtf2-ARE combining capacity by arsenic participates is involved in the regulation of Nrf2 downstream antioxidase gene expression,and this process possibly participates in occurrence and development of coal-burning-borne arsenism and hepatic injury.

Objective To study the expression and enzyme activity of thioredoxin reductase 1 (TrxR1) in liver and peripheral blood of human and rats exposed to airborne arsenic through coal-burning as well as its role in liver injury of coal-burning-borne arsenic poisoning.Methods This study was divided into 2 parts.Part 1 was a population study:133 local residents exposed to airborne arsenic through coal-burning were selected as arsenic exposure groups including a non-patient group (25 cases),no obvious hepatopathy group (38 cases),mild (43 cases) and moderate to severe hepatopathy groups (27 cases) from areas affected by endemic arsenism in Guizhou Province.Thirty-four healthy residents from arsenic not affected areas were selected as controls.Peripheral blood samples were collected from all these people.The expression of TrxR1 mRNA was determined by real-time fluorescence quantitative PCR (qPCR),and enzyme activity of TrxR was tested by visible spectrophotometry.Part 2 was an animal experiment study:Thirty Wistar rats,weighing about 80-100 g,were divided into control group,drinking-waterborne arsenic poisoning group and coal-burning-borne arsenic poisoning group (including low,medium and high arsenic contaminated grain groups) by means of a table of random number according to body mass,6 rats in each group.The control group was fed with normal diet for 3 months; drinking-water-borne arsenic poisoning group and coal-burning-borne arsenic poisoning group were fed with 10 mg/kg As2O3 solution and different concentrations(25,50,100 mg/kg) of arsenic-containing feed,respectively,for 3 months.The expression of TrxR1 mRNA was determined by qPCR; protein expression level of TrxR1 in liver tissue was detected by immunohistochemistry,and enzyme activity of TrxR in serum and liver tissue was tested by visible spectrophotometry.Results The mRNA expressions of TrxR1 in peripheral blood were 1.599 8 (1.128 9-2.156 8),1.469 3 (1.146 1-1.976 3),1.203 6 (0.463 1-1.816 2) and 0.912 3(0.631 8-1.535 0),respectively,among non-patient group,no obvious hepatopathy group,mild and moderate to severe hepatopathy groups.Compared to the control group[1.649 7(1.161 1-2.380 2)],the differences were significant statistically in mild and moderate to severe hepatopathy groups (all P ＜ 0.05).The enzyme activity of TrxR in peripheral blood was (3.12 ± 0.76),(2.81 ± 0.84),(2.52 ± 0.73),(2.42 ± 0.76)U/ml,respectively,in those corresponding groups.Compared to the control group [(3.02 ± 0.70)U/ml],the differences were significant statistically in mild and moderate to severe hepatopathy groups (all P ＜ 0.05).The mRNA expressions of TrxR1 in peripheral blood were 1.05 ± 0.14,1.18 ± 0.18,1.04 ± 0.10 and 0.97 ± 0.13,respectively,among drinking-water-borne arsenic poisoning group,low,medium and high arsenic contaminated grain groups; all of which were lower than that in the control group (1.23 ± 0.15,all P ＜ 0.05) except that of the low arsenic contaminated grain group.The mRNA expressions of TrxR1 in liver tissue were 0.78± 0.10,0.83 ± 0.10,0.79 ± 0.09 and 0.77 ± 0.11,respectively; all of which were lower than that in the control group (0.94 ± 0.12,all P ＜ 0.05).The protein expression of TrxR1 in liver tissue was 310.33 ± 38.81,312.50 ± 23.36,305.67 ± 20.57 and 298.17 ± 23.52,respectively,among the arsenic poisoning groups; all of which were lower than that in the control group (348.50 ± 32.35,all P ＜ 0.05).The enzyme activity of TrxR in serum was (4.22 ± 0.73),(4.86 ± 0.63),(4.04 ± 0.57),(3.73 ± 0.64)U/ml,respectively; all of which were lower than that in the control group [(9.52 ± 1.08)U/ml,all P ＜ 0.05].The enzyme activity of TrxR in liver tissue was (14.82 ± 1.67),(18.76 ± 2.76),(14.90 ± 2.17),(11.55 ± 1.74) U/mg,respectively; all of which were lower than that in the control group [(23.71 ± 3.05)U/mg,all P ＜ 0.05].Conclusion Arsenic aggravates liver injury of coal-burning arsenic poisoning through down-regulating the expressions of TrxR1 mRNA and protein and reducing its enzyme activity as well.

Objective: To investigate the roles of Bcl-xl and Bcl-xs in the development and progression of endometrial carcinoma, and to explore their correlation.Methods: The expression of Bcl-xl and Bcl-xs in 32 cases of endometrial carci-noma, 12 cases of endometrial atypical hyperplasia, 6 cases of endometrial simple hyperplasia and 10 cases of normal en-dometrial tissues were examined by RT-PCR and Western-blot.Results: The expression of Bcl-xl mRNA and protein was significantly higher in endometrial cancer tissues than in normal endometrial tissues (P<0.05), and was statistically associat-ed with the pathological stage of endometrial carcinoma.(F=5.33, P=0.02).The expression of Bcl-xs mRNA and protein in atypical endometrial hyperplasia and endometrial carcinoma tissues were significantlly lower than that in normal endometri-al tissues (P<0.05), which was also associated with clinical stage and lymph node metastasis of endometrial carcinoma (P<0.05).The expression of Bcl-xl was negatively correlated with the expression of Bcl-xs in different endometrial tissues (r=-0.76).Conclusion: The abnormal expression of Bcl-xs and Bcl-xl was a factor for the pathogenesis and development of endometrial carcinoma.The negative correlation between Bcl-xl and Bcl-xs in different endometrial tissues as well as their relative expression ratio may have certain impact on the genesis of endornetrial cancer.

This paper provides a detail review for the Standards in Speech Assessment for Chinese Children,which includes a general framework and its contents.A step-by-step procedure for the speech assessment and therapy has been provided.

In order to promote the integration between disciplines, the convergence between basic course and clinical teaching, increasing students ability including the active learning and life-long learning ability, finding problem and solving problem ability, teamwork spirit and so on. After nearly 3 years preparation, Hebei Medical University successfully carried out the PBL teaching in Seven-year Clinical Medicine Science. Combining with the teaching activities, formative assessment was carried out, and PBL teaching website was established. The reform has already achieved initial results, got good responds from teachers and students. Through the study, it has been confirmed that the PBL teaching method in Hebei Medical University is effective and worthy to reference.

Objective To investigate the relationship between genetic polymorphisms in nucleotide excision repair gene excision repair cross complementing group 6(ERCC6),xeroderma pigmentosum group A(XPA) and coal-burning-borne-arsenism in Guizhou Province.Method ERCC6 A3368G,ERCC6 C-6530G and XPA A23G gene polymorphisms were analyzed by polymerase chain reaction restriction fragment length polymorphism technique(PCR-RFLP) of 205 cases which were chosen as patients with arsenism and 187 residents as control group.Results The distributions of ERCC6 A3368G,ERCC6 C-6530G and XPA A23G in the case group were not statistically significant compared with those of the control group(x2 =3.209,2.963,3.335,all P ＞ 0.05); individuals carrying G allelomorphic gene(AG + GG) had a lower risk than individuals carring A allelomorphic gene(ORadj =0.282,95％CI:0.126-0.628,P =0.002); relationship was not found between single genetic polymorphisms of ERCC6 C-6530G,XPA A23G and coal-burning-borne-arsenism; the risk of arsenism was decreased for individuals carrying the following five genotypes combination:ERCC6 A3368G(AG + GG) genotype and ERCC6 C-6530G CC genotype(ORadj =0.287,95％CI:0.087-0.946,P=0.040); ERCC6 A3368G(AG + GG) genotype and ERCC6 C-6530G(CG + GG) genotype (ORadj =0.226,95％CI:0.077-0.661,P =0.007); ERCC6 A3368G(AG + GG) genotype and XPA A23G AA genotype (ORadj =0.150,95％CI:0.038-0.596,P =0.007); ERCC6 A3368G (AG + GG) genotype and XPA A23G(AG + GG) genotype(ORadj =0.325,95％CI:0.118-0.897,P =0.030) ; ERCC6 C6530G (CG + GG) genotype and XPA A23G AA genotype (ORadj =0.397,95％CI:0.162-0.975,P=0.036).Conclusions Individuals carring ERCC6 A3368G (AG + GG) genotype have a low risk of arsenism.There are five genotypes combination of three gene polymorphisms in two genes,ERCC6 and XPA,which may reduce the risk of coal-burning-borne-arsenism.