Objective To evaluate the expression and significance of differential expression gene Hepsin in the prostate cancer (PC) screened by the cDNA microarray technique.Methods The techniques of semiquantitative RT-PCR and Western blotting were used to detect the mRNA and protein expression of Hepsin. Specimens of 40 cases of PC, 15 benign prostatic hyperplasia (BPH) and 6 normal prostate tissues were examined by the immunohistochemical stain.Results Hepsin was more expressed in PC tissue than normal prostate tissue (P=0.026) and was confirmed by Western blot analysis. Immunohistochemical test confirmed this and demonstrated that Hepsin did not expressed in normal prostate but expressed in PC and BPH and there was a significant difference in Hepsin expression level between PC and BPH tissues ( P=0.000).Conclusion Hepsin high expressed in PC may be a new molecular marker in early diagnosis of PC and a new target for gene therapy of PC.

Four screening methods, colorimetric assay, blood-plate hemolysis method, surface tension activ- ity and oil spreading technique were introduced to isolate strains producing bio-demulsifiers from 6 different bacteria source samples. The results of various screening methods were evaluated in this paper. Seventeen demulsifying strains were obtained, which are qualified in demulsification test of kerosene model emulsions. Among them, 5 strains showed high demulsifying ability, achieving 70% plus demulsifying ratio within 24 hours. Petroleum-contaminated soil, excess sludge from biological process treating oilfield produced water and sludge from municipal wastewater treatment plant were the best among all tested sources. Due to the determination limit, the colorimetric assay and blood-plate hemolysis method are not competent to screen bio-demulsifiers strains. The measurement of surface tension and oil spreading method were easy but accu- rate methods to isolate bio-demulsifiers strains. Although demulsification test of model emulsion is an effec- tive technique to target strains with the capability of breaking emulsions, it is sophisticated and time-con- suming. Thus it is recommended to use surface tension and oil spreading methods in pre-screening and vali- date the results in demulsification test with kerosene model emulsions.

Objectives To screen the differentially expressed proteins in serum of population chronically exposed to arsenic in drinking water,thus to provide candidate protein biomarkers for arsenic exposure and arsenicosis.Methods Subjects were selected from the drinking water type of endemic arsenicosis areas in Shanxi province,China.Demographic characteristics,history of arsenic exposure,cigarette smoking,alcohol drinking,health and other information were collected using questionnaire.The subjects were divided into low-arsenic group (with arsenic in drinking water ＜ 10 μg/L),medium-arsenic group( 10 - 50 μg/L),high-arsenic group( ＞ 50 μg/L),and arsenicosis group(the drinking water with arsenic ＞ 50 μg/L was replaced by low arsenic water ＜ 10 μg/L).The number of cases in each group was 30.The arsenicosis patients were diagnosed according to “Standard of Diagnosis for Endemic Arsenism” (WS/T 211-2001 ).With the principle of informed consent,blood samples were collected.Differentially expressed serum proteins of different arsenic exposure groups and arsenicosis group were screened by two-dimensional differential gel electrophoresis(2-D DIGE),and further identified by mass spectrometry (MS).Results An average of (1299 ± 167) protein spots were identified in 6 gel images and 688 protein spots were discovered repeatedly in at least 5 gels.There were 33 protein spots differentially expressed among low-,medium- and high-arsenic groups P ＜ 0.01).Fifty four protein spots were significantly different among low-,medium-,high-arsenic exposure groups and arsenicosis group(P ＜ 0.01 ).Twenty five protein spots were selected for MS analysis,and13 protein spots were identified.Compared with low-arsenic group,the expressions of apolipoprotein A-Ⅳ,retinol binding protein,and estrogen receptor hypothalamic isoform in medium- and higharsenic exposure groups were down regulated,and the expressions of component 4A and 4B were up regulated.Compared with low-,medium- and high-arsenic groups,the expressions of beta-2-glycoprotein Ⅰ,Keratin 1,hemopexin,complement C1r subcomponent,and ficolin-3 in arsenicosis group were down regulated,and the expressions of pigment epithelial-differentiating factor,alpha-1-microglobulin and carboxypeptidase N catalytic chain were up regulated.Conclusions Chronic arsenic exposure can significantly change population's serum protein expression.Differentially expressed proteins in arsenicosis patients will not decline with the decline of arsenic in a short term.Whether or not the differentially expressed proteins identified in this study can be used as biomarkers for arsenic exposure and arsenicosis needs to be further verified.

Objective To investigate the anti-apoptotic function of clusterin in LNCaP cell line and the role of clusterin antisense oligodcoxynucleotide(AS-ODN)in TNF-?-induced death of LNCaP cell. Methods The wild type LNCaP(group L),LNCaP transfected with the control vector(group M),LNCaP transfected with full-length clusterin expression vector(group A,ie,study group)were cultured.For the de- tection of cytotoxic effect of TNF-?,MTT and ELISA methods were used to determine the cell proliferation and apoptosis of the 3 clones,and the changes of proliferation and apoptosis in A cell after transfection of clusterin AS-ODN were also assessed.Results MTT method showed that the cell proliferation activity(A value)of groups L,M,and A were 0.84?0.03,0.85?0.04,0.95?0.03,respectively;the difference be- tween groups L and M was not significant(P＞0.05);but compared with group A the cell proliferation activ- ity was significantly lower in groups L and M(P＜0.01 for both).ELISA resuhs showed that the A values of groups L,M,and A were 0.59?0.04,0.62?0.03,0.33?0.04,respectively;the difference between groups L and M was not significant(P＞0.05);but compared with group A,the apoptosis rates were significantly higher in groups L and M(P＜0.01 for both).In group A,A values of cell proliferation activity in subgroups control,AS-ODN,TNF-?,TNF-?+AS-ODN were 1.30?0.03,1.25?0.03,0.99?0.03,0.80?0.03, respectively;the differences between each group were significant(P＜0.05 for all).And the A values of cell apoptosis in the above 4 groups were 0.02?0.00,0.21?0.02,0.63?0.07,1.16?0.04,respectively,the differences between each group were significant(P＜0.01 for all).Conclusions Stable transfection and subsequent expression of clusterin result in resistance to the cytotoxic effect of TNF-?.Transfection with clus- terin AS-ODN enhances cytotoxic effect of TNF-?in A cells.These results suggest that clusterin plays an im- portant role in anti-apoptotic function in LNCaP cell line.

The mechanism of apoptosis induced by SIRT1 deacetylase inhibitors in both human breast cancer MCF-7 and MCF-7 doxorubicin-resistant cells was studied. MTT assay was used to detect growth-inhibitory effect on the cells. Protein expression was detected by Western blotting. Chromatin condensation was detected by a fluorescent microscope after Hoechst 33342 staining. Cell cycle distribution was analyzed with flow cytometry. Apoptotic cells were detected with Annexin V staining. Nicotinamide (NAM) and Sirtinol, two SIRT1 deacetylase inhibitors, exhibited the similar growth-inhibitory effects on MCF-7/DOX cells and MCF-7 cells, but no potentiation of DOX activities. The arrest at G2/M phase was detected by flow cytometry in both MCF-7 and MCF-7/DOX cells after NAM treatment. Activation of caspase pathway in MCF-7 cells, such as the cleavages of PARP, caspase-6, -7, -9, were observed after exposure to NAM 50 mmol x L(-1), accompanied by the occurrence of chromatin condensation and Annexin V positive cells. However, the cleavages of PARP, caspase-6 and -7 in MCF-7/DOX cells delayed after exposure to NAM for 24 h and obviously increased at 48 h with appearance of chromatin condensation and Annexin V positive cells. SIRT1 deacetylase inhibitors show no cross resistance to MCF-7 drug-resistant cells, and the similar growth-inhibitory actions of them to MCF-7 sensitive and drug-resistant cells by which it is mediated by activation of apoptotic caspase pathway.
Apoptosis ; drug effects ; Benzamides ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Caspases ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Enzyme Inhibitors ; pharmacology ; Female ; Humans ; Naphthols ; pharmacology ; Niacinamide ; pharmacology ; Sirtuin 1 ; antagonists & inhibitors

OBJECTIVETo study the risk factors related to the survival rate, recurrence and metastasis of malignant fibrous histiocytoma of bone.

METHODSFrom July 1997 and July 2010, 56 patients with malignant fibrous histiocytoma of bone were treated. Univariate and multivariate analysis were performed to determine the probable risk factors including gender, age, tumor location, tumor size and so on.

RESULTSForty-four cases were followed up ranged from 2 weeks to 78 months (medium 33.3). The 5-year overall survival rate was 50.1%, local recurrence rate 40.9% with a median time of 12 months (3 to 60 months) and metastatic rate 27.5% (11/40) with a median time of 6.5 months (2 to 23 months). Univariate analysis indicated that gender, condition of presentation (primary case or recurrence case), tumor location, surgical margin and surgical stage were significantly related to survival rate (P < 0.05), and tumor location and surgical margin were related to local recurrence rate (P < 0.05), and important vessel or nerve invasion was related to metastatic rate (P < 0.05). Multivariate analysis showed that surgical margin and surgical stage were independent risk factors for survival rate, of which surgical margin was the independent risk factor for recurrence rate.

CONCLUSIONSSurgical margin and surgical stage are independent risk factors for survival rate, of which surgical margin is the independent risk factor for recurrence rate.

Adolescent ; Adult ; Aged ; Bone Neoplasms ; diagnosis ; pathology ; Female ; Histiocytoma, Malignant Fibrous ; diagnosis ; pathology ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Recurrence, Local ; pathology ; Prognosis ; Retrospective Studies ; Survival Rate ; Young Adult

OBJECTIVETo study the clinical features and surgical treatment of osteoid osteoma and improve the diagnostic therapeutic level.

METHODSClinical data of 35 patients (25 males and 10 females) with osteoid osteoma diagnosed and treated between January 1997 to October 2009 were retrospectively reviewed. The average age was 21 years (ranged, 6 to 49 years). The average interval time between onset of symptoms and diagnosis was 12 months (ranged, 1 to 24 months). The most common sites were the tibia (13 patients) and the femurs (7 patients). The most common presenting complaints for patients with osteoid osteoma was pain which usually responded to NSAIDs and was generally more severe at night. The imaging manifestations revealed a circular or oval nidus. All the patients underwent surgical treatment. The tumors were treated with curettage or excision with autograft or allograft in 19 patients, simple surgical resection in 9 patients, curettage or excision with autograft or allograft and fixation in 7 patients.

RESULTThe mean follow-up period was 49 months (ranged,2 months to 12 years). The symptom of pain disappeared after operation. There were no evidence of recurrence. Tibial pathological fracture happened in one patient 4 months postoperatively, and the patient got healing after plate-screw internal fixation. One patient with sinus formation 5 years postoperatively got wound healing after sinus resection, intramedullary nail removal and debridement.

CONCLUSIONAccording to the typical clinical presentation, radiographic findings, the diagnosis of osteoid osteoma is not difficult. Once the diagnosis is confirmed, the operation should be carried out as early as possible to relieve the symptoms, improve the quality of life and prevent long-term complications.

Adolescent ; Adult ; Bone Neoplasms ; diagnosis ; surgery ; Child ; Female ; Humans ; Male ; Middle Aged ; Osteoma, Osteoid ; diagnosis ; surgery

Objective To investigate the relationship between drinking-water type of endemic arsenicosis and adult carotid artery atherosclerosis. Methods In 2009, 285 participants aged over 40 from drinking-water type of endemic arsenism areas and 293 residents aged over 40 from control areas were investigated in Yingxian county,Shanxi province. Portable-type B mode color ultrasound was used to examine the carotid artery of all participants.The carotid atherosclerosis were diagnosed and graded through the ultrasonograms. Content of water arsenic and hair arsenic of 10 people randomly selected in every villages were detected. Results A total of 5 villages with drinkingwater type of endemic arsenicosis as observation group and 5 villages without drinking-water type of endemic arsenicosis as control group were investigated. The prevalence rates of adult carotid atherosclerosis within observation group were 35.09%(20/57), 55.74%(34/61), 38.46%(20/52), 36.51%(23/63) and 46.15%(24/52), respectively,and standardized prevalence rates were 32.5%, 33.8%, 34.9%, 46.2% and 47.3%, respectively and the prevalence rates of adult carotid atherosclerosis within control group were 18.18%(10/55), 30.77%(16/52), 20.00%(10/50),18.67% (14/75) and 21.31% ( 13/61 ), respectively; the standardize prevalence rates were 22.4%, 17.7%, 10.7%,24.6%, 18.9%, respectively. The standardize prevalence rates were higher in observation group [39.50%(113/285) ]than that in control group[39.50%(113/285), T = 26, P ＜ 0.01 ]. The severity of adult carotid atherosclerosis (composition of 4 - 7 scores ) was compared between observation group [ 17.70%(20/113 )] and control group [ 14.06% (9/64) ], and the difference was insignificant(x2 = 0.26, P ＞ 0.05). Conclusions The prevalence rate of carotid atherosclerosis in drinking-water type of endemic arsenicosis areas is higher than that of the control areas.The study provides evidence that arsenic poisoning can cause atherosclerosis.

ObjectiveTo observe the changes of fluoride content in serum,bone and urine after rats were exposed to single fluoride,single aluminum or fluoride combined with aluminum and to investigate the effects of different doses of aluminum on fluoride accumulation and excretion in rats.Methods Male Wistar rats were randomly divided into 9 groups based on 3 × 3 factorial design.Different doses of fluoride(NaF,0,50,200 mg/L)and(or) aluminum(AlCl3,0,100,200 mg/L) were administered to rats in each group by drinking water.The rats took food and water ad libitum during the experimental period.After feeding for 18 weeks,rats with obvious dental fluorosis were determined as successful establishment of animal model.The fluoride content in the serum,bones and urine were measured.Results Fluoride affected the fluoride content in serum,bones and urine(F=166.74,577.81,160.96,all P ＜ 0.01 ).The interaction of fluoride and aluminum on serum,bone and urinary fluoride were statistically significant (F =7.95,5.13,6.94,all P ＜ 0.01 ).When the fluoride level was 50 mg/L,the serum fluoride contents were [ (0.08 ± 0.03) and (0.08 ± 0.02) mg/L] in the aluminum levels of 0 and 100 mg/L groups,which was higher than that of the aluminum level of 200 mg/L group[ (0.04 ± 0.01)mg/L,F=7.14,5.78.all P＜ 0.05].The bone fluoride content in the 0 mg/L aluminum level group[ (1996.53 ± 383.73) mg/kg] was higher than that of the 100 and 200 mg/L groups[(1252.51 ± 189.08),( 1160.63 ± 129.63) mg/kg,F=20.54,24.56,all P ＜ 0.01 ].When the fluoride level was 200 mg/L,the bone fluoride contents were decreased with the increasing doses of aluminum[ (4668.70 ± 887.67),(3920.30 ± 528.31 ),(3297.64 ± 396.04) mg/kg].Between any two groups,the differences were statistically significant (F =15.59,52.31,14.38,all P ＜ 0.01 ).When the fluoride level was 50 mg/L,the urinary fluoride content in the 0 mg/L aluminum level group[ (34.054 ± 9.30)mg/L] was higher than that of the 100,200 mg/L groups[( 14.81 ± 6.32),(14.67 ± 3.42) mg/L,F =25.30,24.32,all P ＜ 0.01 ].When the fluoride level was 200 mg/L,the urinary fluoride contents in the 0,100 mg/L aluminum level groups[ (57.14 ± 21.38),(51.75 ± 8.39)mg/L] were higher than that of the 200 mg/L group[(34.839 ± 9.30) mg/L,F=30.04,20.31,all P ＜ 0.01 ].ConclusionsAluminum is an antagonist of fluoride.The antagonism could be enhanced as the dose of aluminum increased.In this study,aluminum could effectively counteract the absorption of fluoride in rat model when the ratio of fluoride to aluminum is 1 ∶ 2.

Breeding is not only an important area of medicinal plants research but also the foundation for the superior varieties acquirement of medicinal plants. The rise of modern biotechnology provides good opportunities and new means for medicinal plants breeding research in China. Biotechnology shows its technical advantages and new development prospects in breeding of new medicinal plants varieties with high and stable yield, good quality, as well as stress-resistance. In this paper, we describe recent advances, problems, and development prospects about the application of modern biotechnology in medicinal plants breeding research in China.
Biotechnology ; methods ; Breeding ; China ; Plants, Medicinal ; genetics ; Research ; Tissue Culture Techniques