Objective To investigate DNA methylation in the promoter region,mRNA transcription and protein expression of glutathione-S-transferases-P1 (GSTP1) gene and their relation with arsenism.Methods In endemic coal-pollution-borne arsenism area,Jiaole village of Xinren county,Guizhou province,according to the diagnostic criteria of endemic arsenism(WS/T 211-2001),123 cases with endemic arsenism were selected and divided into three groups (mild arsenism group:42 cases,moderate arsenism group:41 cases and severe arsenism group:40 cases).Forty seven residents were selected as controls in a village about 12 km away from the endemic arsenism area.With the informed consent principle,peripheral blood of all respondents was collected in order to analyze DNA methylation and check mRNA.DNA methylation of GSTP1 gene promoter region in peripheral blood was assayed by PCR,and GSTP1 mRNA expression was assayed using real-time quantitative PCR.In addition,other cutaneous specimens originated from 53 cases with arsenism that accepted surgical treatment voluntarily were taken.Of these specimens,general pathological changes were 28 cases,precancerous 20 cases and cancerous 5 cases.Skin tissues of 15 cases of non-tumor surgery patients without abnormal pathological changes were as control group.GSTP1 protein expression in the skin tissue was detected using immunohistochemistry (IHC).Results Among different groups of arsenic poisoning,the positive rate of DNA methylation of GSTP1 gene was 28.57％(12/42) in the mild group,57.10％ (23/41) in the moderate group and 65.00％ (26/40) in the severe group.Compared with the control group (6.38％,3/47),the difference was statistically significant (x2 =7.792,26.000,33.412,all P ＜ 0.01).Among different groups of arsenic poisoning diagnosed by dermapathology,the positive rate of DNA methylation of GSTP1 gene was 21.43％(6/28) in the general pathological change group,50.00％(10/20) in the precancerous group and 80.00％(4/5) in the cancerous group.Compared with the control group(6.67％,1/15),the difference was statistically significant (x2 =3.562,7.468,10.756,all P ＜ 0.05).It showed that the positive rate of DNA methylation of GSTP1 gene increased with aggravation of the disease and dermatic lesion of arsenism (tendency x2 =38.239,x2 =13.659,all P ＜ 0.01).Compared with the control group(0.184 26),the expressions of GSTP1 mRNA in peripheral blood in moderate (0.087 77) and severe arsenic poisoning groups (0.056 93) were significantly reduced(all P ＜0.01),and that of severe group was significantly lower than that of the moderate group (P ＜ 0.01) ; compared with the control group(0.338 45) and the general lesion group(0.276 74),GSTP1 mRNA expression was significantly reduced in precancerous lesion group(0.104 81) and cancerous group(0.043 70),in which the cancerous group was significantly lower than that of the precancerous lesions.The difference of skin tissue GSTP1 protein expression rate between groups was statistically significant (x2 =20.948,P ＜ 0.05),in which the difference between the precancerous lesion group(65.00％,13/20),the cancer group (40.00％,2/5) and the control group(100.00％,15/15)was statistically significant (x2 =12.183,11.778,P ＜ 0.01).Spearman correlation analysis showed that the degree of skin lesion and the level of GSTP1 protein expression was negatively correlated (r =-0.520,P ＜ 0.05).Groups were divided according to DNA methylation of GSTP1 gene,and the mRNA and protein expression of GSTP1 in methylation group(0.038 40,57.14％) was significantly lower compared with that of unmethylated group(0.187 07,95.74％; Z =9.032,x2 =23.134,all P ＜ 0.01).Conclusions Arsenism may lead to DNA methylation of human GSTP1 gene promoter region,thereby inhibiting expression of mRNA and protein.GSTP1 gene plays an important role in arsenism or carcinogenic process.

Objective To evaluate the operative treatment of nonunion of humeral shaft fracture with in- ternal plate fixation and autogenous cancellous hone graft.Methods Forty-one cases of nonunion of humeral shaft fracture operatively treated from February 2002 to December 2004 were analyzed retrospectively.There were 32 males and nine females.Their average age was 37.5 years(range,17 to 67 years).Sixteen nonunions were defined as hypertrophic and 25 as atrophic.We followed all the patients and obtained their complete medical information. Results Our average follow-up was 22.6 months(range,8 to 42 months).Forty fractures(97.6%)were united within an average of 5.8 months(range,3 to 12 months).Complications included iatrogenic radial nerve injury in three patients,wound infection in one patient and fracture nonunion in one patient.At the final follow-up,shou]der and elbow functions were found to be satisfactory.Conclusion Open reduction and plate internal fixation sup- plemented with autogenous cancellous bone graft is an effective treatment for nonunion of humeral shaft fracture.

Objective To explore the possible mechanisms of Arsenic Trioxide in treating rheumatoid arthritis(RA)by observing the changes of HE staining and NF-KB expression as well as the apoptosis of syn- oviocytes in adjuvant-induced arthritis rats.Methods After the animal model was set up on Wistar rats sue- cessfully,they were randomly divided into AA model group and arsenic trioxide treatment group.The treat- ment group were injected with 4 mg'kg~(-1)9?d~(-1)arsenic trioxid fluid for 7 days.All of the rats were killed 3 days after the complete of injections.The joint specimens were exposed,fixed,decalcified,wrapped and cut into slices.All slices were examined by HE stain and immunohistological evaluation.Results HE staining showed that when compared with the normal control group,the layers of synoviocytes of the AA group were increased to 6-8,and the arrangement of synoviocytes was disordered and heavy inflammatory cell infiltration were found in the AA group.In the arsenic trioxide treatment group,the layers of synoviocytes increased to 3～4,and medi- um amount of inflammatory cell infiltration were found.The intensity of synovial NF-kB(p65)positive stain in AA model group was significantly higher than that in the normal control group.The synovial expression and ac- tivation of NF-kB in the treatment group were decreased markedly,and did not return to normal level.The average gray scale calculation showed that there were significant differences between the three groups(P

Objective Construct the eukaryotic expression vector of inhibitory member of the ASPP family (iASPP) and trans-fect it into colon carcinoma cell lines SW480 and Lovo by liposome .Then observe the expression of iASPP and detect the cell apop-tosis by flow cytometry .Methods The amplified PCR product was digested and inserted into pMD19-T simple vector and sub-cloned into eukaryotic expression vector pcDNA3 .1(+ ) .The recombinant eukaryotic expression plasmid pcDNA3 .1(+ )-iASPP was transfected into colon carcinoma cell lines SW480 and Lovo by liposome ,the iASPP expression was analyzed by RT-PCR .The cell apoptosis was detected by FCM .Results The eukaryotic expression plasmid pcDNA3 .1(+ )-iASPP was constructed success-fully ,the gene squence of iASPP was consistent with that reported (gi 60457962) in GenBank .The mRNA expression levels of iASPP gene of SW480 and Lovo cell lines which transfect the positive plasmid were increased ,and the cell apoptosis rates were de-creased .Conclusion We successfully constructed the recombinant expression plasmid pcDNA 3 .1(+ )-iASPP ,and the plasmid were successfully expressed in colon carcinoma cell lines SW 480 and Lovo ,the cell apoptosis rates of those cell lines were decreased .These facts indicated that reducing the high expression of iASPP may be a new strategy to renew the abilities of P 53 tumor suppressor .

Objective To analyze epidemiological characteristics of elderly cases with influenza A (H1N1) in Beijing. Methods Descriptive epidemiological methods were used to describe epidemiological characteristics of elderly cases with 2009 influenza A (H1N1) in Beijing. Results The 321 laboratory-confirmed elderly cases with influenza A (H1N1) were reported in Beijing, and the morbidity was 13.2/100 000. The peak of infection occurred during November and December, the cases in this period accounted for 84.7% of the whole year, and 53.0% of them were reported in suburb areas, with the highest morbidity (19.2/100 000) in people beyond 85 years, and the morbidity increased with age (x2 = 7.24, P＜0.01). The mild cases accounted for 63.6 %, severe and critical cases accounted for 36.4%. No significant difference was found between severity and BMI (x2=8.14, P=0.52). Severity was associated with number of chronic diseases (x2= 123.0, P＜0. 01). Conclusions The H1N1 morbidity and proportion of severe cases are high among the elderly in Beijing, more attention should be paid to this population for influenza A (H1N1) prevention and control.

OBJECTIVETo investigate the effect of oligochitosan in promoting intestinal absorption of protoberberine alkaloids in extracts from Corydalis saxicola total alkaloids.

METHODThe in vitro single-pass intestinal perfusion model in rats was established to study the changes in absorption kinetic parameters of dehydrocavidine, berberine hydrochloride and palmatine chloride in C. saxicola total alkaloids after the addition of different concentrations oligochitosan and evaluate the effect of oligochitosan in promoting intestinal absorption of the drugs.

RESULTThe concentration of oligochitosan had different effects on the absorption rate constant (Ka) and apparent permeability coefficient (Peff) of the three active component in rat intestines. Ka and Peff in 0.5% oligochitosan group significantly increased, indicating a stronger effect in promoting the absorption.

CONCLUSIONOligochitosan has a certain effect in promoting the intestinal absorptions of protoberberine alkaloids in C. saxicola total alkaloids.

Animals ; Berberine Alkaloids ; administration & dosage ; pharmacokinetics ; Chitin ; administration & dosage ; analogs & derivatives ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo determine the expression of Skp2 in different prostate cancer (PCa) cell lines and tissues, and explore its influence on the androgen receptor (AR) signaling pathway and development of castration-resistant prostate cancer (CRPC).

METHODSThe expression levels of Skp2 and AR in different PCa cell lines were detected by Western blot. After knockdown of Skp2 in the C4-2 and 22RV1 cells transfected with shRNA, the expressions of AR and P27 were determined and the activity of ARR3-Luc measured by dual-luciferase reporter gene assay following treatment with dihydrotestosterone (DHT). The expressions of AR and Skp2 in human naïve PCa or CRPC specimens were detected by immunohistochemical staining followed by analysis of their differences and correlation.

RESULTSThe Skp2 protein expression level was significantly higher in the C4-2 or 22RV1 cells than in the LNCaP cells. DHT treatment increased the expression of Skp2 in the C4-2 cells, but knock-down of Skp2 significantly up-regulated the expression of the well-known downstream protein P27 and down-regulated that of AR. Consistently, DHT treatment increased the activity of ARR3-Luc, while knockdown of Skp2 remarkably decreased it in the C4-2 and 22RV1 cells (P < 0.05). In addition, significantly higher expressions of Skp2 and AR were observed in the CRPC than in the naïve specimens (P < 0.05), with a positive correlation between the two proteins (r = 0.658 1, P < 0.05).

CONCLUSIONSkp2 can enhance the expression and transcription activity of the AR protein in CRPC cells or tissues and is promising to be a critical molecular therapeutic target.

Androgens ; pharmacology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Disease Progression ; Gene Knockdown Techniques ; Humans ; Male ; Neoplasm Proteins ; genetics ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; metabolism ; Receptors, Androgen ; genetics ; metabolism ; S-Phase Kinase-Associated Proteins ; physiology ; Transcriptional Activation ; Up-Regulation

Objective To study morphological changes of rabbit artery endothelial cell injury and atherosclerosis caused by high fluoride and the role of selenium. Methods Twenty healthy male New Zealand white rabbits, body weight (2.0 ± 0.5)kg, were randomly divided into control group(drinking deionized water, fed basic diet), fluoride group(drinking fluoride 100 mg/L deionized water, fed basic diet), selenium group(drinking selenium 1 mg/L deionized water, fed basic diet), fluoride plus selenium group(drinking fluoride 100 mg/L deionized water, selenium 1 mg/L of deionized water, fed basic diet). The experimental period was 6 months. At 0, 3, 6 months of the experiment, serum fluorine and selenium levels were determined. At the end of the experiment,thoracic aorta was collected to observe its pathology and ultrastructural changes. Results Serum fluoride was significantly higher at the 3rd and the 6th month of experiment(all P ＜ 0.01 ) in fluoride group[ (0.589 ± 0.146),(0.772 ± 0.175)mg/L] and fluoride plus selenium group[ (0.502 ± 0.094), (0.693 ± 0.158)mg/L] than in control group[ (0.174 ± 0.002), (0.208 ± 0.031 )mg/L] and serum fluoride was significantly higher at 6 months than at 3 months(P ＜ 0.05 ) in fluoride group. Serum selenium was significantly higher at the 3rd and the 6th month of experiment (all P ＜ 0.01 ) in selenium group[ (0.252 ± 0.022), (0.319 ± 0.052)mg/L] and fluoride plus selenium group[ (0.239 ±0.016), (0.294 ± 0.018)mg/L] than in control group[(0.135 ± 0.014), (0.167 ± 0.019)mg/L], and serum selenium was significantly higher at the 6th month than at 3rd month of experiment in selenium group(P ＜ 0.05). Endothelial cell apoptosis indices were (4.92 ± 1.32)%, (30.30 ± 6.80)%, (6.57 ± 2.14)% and (14.29 ± 2.99)%, respectively in control group, fluoride group, selenium group and fluoride plus selenium group. Their main effect of fluorine and selenium was statistically significant (F = 106.833,20.082, all P ＜ 0.01 ). There were antagonistic effect between fluoride and selenium(F = 30.402, P ＜ 0.01 ). Pathological changes of rabbit aortic endothelial cells in fluoride group included endothelial with attached fibrin and red blood cells, and structural of the cells changed, with serious vascular injury; in fluoride plus selenium group apoptosis of endothelial cells decreased, with reduced number of attached red blood cells and fibrin, endothelial cell structure normal, the extent and scope of vascular damage significantly reduced. Conclusions Appropriate amount of selenium inhibits the apoptosis of endothelial cells induced by high fluoride, reduces aortic structural damage caused by high fluoride, and maintains the integrity of endothelial cells, thereby antagonizes the vascular damage and atherosclerosis induced by high fluoride.

Objective To investigate the value of endovascular surgery for the treatment of thromboangitis obliterans (TAO). Methods Sixteen patients (18 limbs) with thromboangitis obliterans were treated by percutaneous transluminal plasty (PTA). Results Angiographic success on the limb basis was achieved in 15 of 18 treated limbs and the initial technical success rate was 83.34%. Doppler anklebrachial index (ABI) increased from 0.33±0.16 to 0.79±0.23 one week after PTA. The follow-up time was 2～24 months, the average time was 10.84 months. The 3-month, 12-month accumulated primary patency rate were 81.33% and 60. 23% respectively. Conclusion PTA is effective primary invasive treatment for thromboangitis obliterans yielding acceptable primary clinical success with a low complication rate and resulting in moderate long-term clinical patency and a high limb salvage rate.

Objective To evaluate the development of fetal cerebellar vermis using 3D transabdominal ultrasound,and provide evidence for prenatal screening fetal cerebellar vermis anomalies.Methods Totally 387 normal fetuses at 20～36 gestation weeks were examined by three-dimensional extended imaging(3DXI) to observe the fetal cerebellar vermis.The width,anteroposterior diameter,craniocaudal diameter of the cerebellar vermis,the angle between brain stem and vermis,a ratio between the area of anterior vermis and posterior vermis were measured,and the relationship between the gestational age and parameters mentioned above were analyzed.Results The multi slice view of 3DXI and 3D reconstructed sagittal section well evaluated the integrity of vermis morphous and vermis size,identified characteristic signs of vermis:the fourth ventricle apex and vermis crack.In normal fetus,brain stem and vermis were almost parallel,the angle between them was 3.97°±1.65°.There was no significant correlation between the angle and the gestational age.The area of the anterior vermis was smaller than that of posterior vermis,with a ratio of 0.76±0.06,which was also not related to gestational age.The width,anteroposte rior diameter and craniocaudal diameter of the cerebellar vermis were positively correlated with gestational age.Conclusions The multi slice view of 3DXI and 3D reconstructed sagittal section should help evaluate the development of the cerebellar vermis,accurately show the cerebellar vermis and its surrounding strutures,and provide a new way to evaluate the fetal cerebellar vermis.