Objective To develop the detection method of common fungal pathogens by general primer PCR.Methods The primers were designed from the target genes of 5.8S rDNA and 28S rDNA of fungi,and the specificity and sensitivity were observed.Results The general primer PCR can be used to detected C.albicans,C.parapsilosis,C.krusei,C.glabrata,C.tropicalis,C.neoformans,A.fumigatus,A.flavus,A.nidulans,A.niger,C.carrionii,P.verrucosa,S.schenckii,F.pedrosoi,T.rubrum,T.mentagrophytes,M.gypseum,M.canis,E.floccosum,M.racemosus,the sensitivity was 15 pg/ml of DNA.Conclusion The general primer PCR can be used to detect common fungal pathogens.

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

0 05) between the two groups in the patient′s age, pregnancy age, parities, the cavity volume of uterine, hemoglobin and blood platelet count The bleeding volume was (51 6?17 2) ml for the treated group and (63 3?17 1) ml for the control group ( P 2 months It could decrease the volume of bleeding during the operation and benefit for patients recovering

Objective To study the content difference of naringin in Fructus Aurantii Immaturus of different specifications.Methods HPLC was performed on the Merck- Lichrospher RP-C18(4.6 mm? 250 mm, 5 ? m) with temperature of 35 ℃ .The chromatographic conditions for naringin:the mobile phase consisting of acetonitrile-0.3 % Phosphoric acid(20 ∶ 80), at the flow rate of 1 mL/min, and the detection wavelength at 283 nm. Results The content of naringin increased with the diameter of Fructus Aurantii Immature slices.Conclusion It is suggested that there exists significant differences in intrinsic quality among Fructus Aurantii Immaturus of different specifications.

Objective To study the relationship between the invasion of pituitary adenomas and expressions of MMP-9,Ki-67 and nm23. Methods The expressions of MMP-9,Ki-67 and nm23 were examined by immunohistochemical Elivision methods in 78 cases of pituitary adenomas. Of them,40 cases were invasive pituitary adenomas and, 38 cases were noninvasive pituitary adenomas. Results The expression levels of MMP-9 and Ki-67 in the invasive pituitary adenomas were significantly higher than those in the noninvasive ones (P

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 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 expression of Zbtb7 gene in Phorbol esters(PMA) induced differentiation of leukemia cell lines.Methods 50 nmol/L PMA was used to induce the differentiation of U937 and K562 cell lines.The expression of Zbtb7 was detected by real-time PCR in the cells after induction on day 0,1,3 and 5,respectively. Results The expression of Zbtb7 in leukemia cell lines was markedly enhanced after treated by PMA(P

Objective To explore the effects of low frequency electric stimulation at bilateral mastoid processes on motor function and cerebral blood flow of children with cerebral palsy (CP). Methods Ninety children with CP were randomly divided into two groups: an electric stimulation group (group 1) and an conventional rehabilitation control group (group 2), thirty healthy children served as the normal control group. The children of group 1 were treated with FES in addition to the routine rehabilitation treatment. The children of group 2 were given of the routine rehabilitation treatment only. All the patients were treated successively with the above protocol for 3 months. The clinical effect and motor development were evaluated with the gross motor function measure (GMFM), and the blood flow velocities of anterior cerebral artery (ACA), middle cerebral artery (MCA), posterior cerebral artery (PCA) were measured by transcranial doppler (TCD) ultrasound before and after treatment. Results In group 1, significant or some improvement were achieved in 19 and 24 CP children, respectively, with an effective rate of 95.6% . In group 2, significant or some improvement were achieved in 10 and 27 CP children, respectively, with an effective rate of 82.2%. There was significant difference between the two groups with regard to the significant effective rate (P