OBJECTIVETo observe the effect of single herb pilose antler (PA) on the expression of Smad2 and Smad3 in the cartilage of osteoarthritis (OA) rats.

METHODSOne hundred 3-month old female healthy SD rats, (200 +/- 20) g, were recruited and routinely fed for 1 week. They were randomly divided into 5 groups, i.e., the low dose PA group, the high dose PA group, the normal saline control group, the model group, and the normal control group, 20 in each group. The model was prepared using classic Hulth method except the normal control group. After 6-week modeling, the model was confirmed successful by pathologic observation. PA at 0.021 g/100 g and 0.084 g/1 00 g was given by gastrogavage to rats in the low dose PA group and the high dose PA group respectively. Normal saline was administered to those in the normal saline control group. No treatment was given to rats in the normal control group and the model group. Bilateral knee cartilages were harvested at week 2,4, and 6. mRNA and protein expressions of Smad2 and Smad3 were detected by immunohistochemical assay, fluorescent quantitative PCR, and Western blot.

RESULTSOA model was successfully prepared by pathological observation. Results of immunohistochemical assay showed that Smad2 and Smad3 expressed extensively in the cartilage, and located inside the chondrocyte membrane. Compared with the model group, mRNA expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 2, 4, and 6, showing statistical difference (P < 0.05). Compared with the same group at week 4 after gastrogavage, mRNA expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.05). Compared with the model group, protein expression of Smad2 and Smad3 obviously increased in the chondrocytes of the low dose PA group and the high dose PA group at week 2 and 4, showing statistical difference (P < 0.01). Compared with the same group at week 2 after gastrogavage, protein expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 4, showing statistical difference (P < 0.01). Compared with the same group at week 4 after gastrogavage, protein expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.01).

CONCLUSIONS(1) The pilose antler could repair cartilages by regulating mRNA and protein expressions of Smad2 and Smad3. (2) Up-regulating mRNA and protein expressions of Smad2 and Smad3 might be one of important mechanisms for the pathogenesis of OA.

Animals ; Antlers ; chemistry ; Cartilage ; cytology ; metabolism ; Chondrocytes ; drug effects ; metabolism ; Female ; Medicine, Chinese Traditional ; Osteoarthritis ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Smad2 Protein ; metabolism ; Smad3 Protein ; metabolism

OBJECTIVETo investigate the expression of heat shock protein 90 (HSP90) on the cell surface of highly invasive human prostate cancer cells PC3 and its possible molecular mechanisms of its effect on cell invasion through analyzing FAK/Src signaling pathway.

METHODSThe expression of cell surface HSP90 on PC3 cells was studied by immunofluorescence staining and surface biotinylation assay respectively. A specific HSP90 antibody was used to inhibit the cell surface HSP90. In vitro cell invasion was assessed by modified Boyden chambers. Phosphorylated FAK on tyr 397, 576, 577 and 925, and phosphorylated c-Src on tyr 416 were examined by Western blot assay. The association between FAK and c-Src was analyzed by immunoprecipitation. The effects of FAK knockdown by siRNA or Src kinases inhibitor PP2, with or without anti-HSP90 antibody, on PC3 cell invasion were also evaluated.

RESULTSA pool of HSP90 was detected on the cell surface of PC3 cells. A specific HSP90 antibody significantly retarded tumor cell invasion. Concomitant with this finding, targeting cell surface HSP90 significantly inhibited the phosphorylations of FAK and c-Src, and also the interactions between FAK and c-Src. FAK knockdown or PP2 dramatically suppressed cell invasion, however, anti-HSP90 antibody didn't further inhibit cell invasion.

CONCLUSIONSCell surface HSP90 promotes human prostate cancer cell invasion through a FAK/c-Src signaling, with may be a novel therapeutic target against metastatic tumors.

Antibodies ; pharmacology ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Focal Adhesion Protein-Tyrosine Kinases ; genetics ; metabolism ; Gene Knockdown Techniques ; HSP90 Heat-Shock Proteins ; immunology ; metabolism ; Humans ; Male ; Neoplasm Invasiveness ; Phosphorylation ; Prostatic Neoplasms ; metabolism ; pathology ; Pyrimidines ; pharmacology ; RNA, Small Interfering ; genetics ; Signal Transduction ; Transfection ; src-Family Kinases ; antagonists & inhibitors ; metabolism

Objective To provide a scientific basis for hospitalized pa-tients receiving warfarin anticoagulant therapy.Methods Collecting 136 medication records of inpatients that have used warfarin in cardiology wards of our hospital from January 2012 to June 2013 , and analyzing the correlation of drug interactions and pathological factors with warfarin effi-cacy and adverse reaction.Results and Conclusion Moxifloxacin , acarbose , metformin and methylprednisolone are probably effect on warfa-rin treatment -related indicators.Infection and bleeding adverse effect prolonged hospitalization days.Warfarin therapy were influenced by drug interactions and pathological factors.

Objective To investigate the distribution of related single nucleotide polymorphisms (SNPs) of drug transporters in healthy Chinese Han population.Methods 10 specific SNPs of 6 drug transporters,including organic anion transporting polypeptide 1B1 (OATP1B1),OATP1B3,organic cation transporter 1 (OCT1),P-glycoprotein (P-gp),multidrug resistance protein 2 (MRF2) and breast cancer resistance protein (BCRP),were determined in healthy Chinese Han volunteers by the established rapid polymerase chain reaction (PCR) methods and sequencing.Gene frequencies were calculated and compared with other populations in the 1000 genomes project.Results The minor allele frequencies of OATP1B1 388A ＞ G,OATP1B1 521T ＞ C,OATP1B3 699G ＞ A,OCT1 181C ＞ T,OCT1 1393G ＞ A,OCT1 1258delA,MDR1 3435T ＞ C,MRP2-24C ＞ T,BCRP 421C ＞ A and BCRP 376C ＞ T were 26.97％,6.25％,23.91％,0,0,0,38.47％,18.52％,31.97％ and 1.48％,respectively.By comparing the distribution of allele frequencies between test group and Mrican,the frequencies of the OATP1 B3 699G ＞ A variant alleles were 76.09％ and 35.63％,respectively;the frequencies of the MDR1 3435T ＞ C variant alleles were 61.53％ and 85.02％,respectively;the frequencies of the MRP2-24C ＞ T variant alleles were 18.52％ and 3.10％,respectively;the frequencies of the BCRP 421C ＞ A variant alleles were 31.97％ and 1.29％,respectively.And these differences were statistically significant (P ＜ 0.05).By comparing the distribution of allele frequencies between test group and American,the frequencies of the OATP1B1 388A ＞ G variant alleles were 26.97％ and 47.26％,respectively;the frequencies of the BCRP 421C ＞A variant alleles were 31.97％ and 14.12％,respectively.And these differences were statistically significant (P ＜ 0.05).By comparing the distribution of allele frequencies between test group and European,the frequencies of the OATP1B1 388A ＞ G variant alleles were 26.97％and 40.26％,respectively;the frequencies of the OATP1B1 521T ＞ C variant alleles were 6.25％ and 16.10％,respectively;the frequencies of the OCT1 181C ＞ T variant alleles were 0 and 6.26％,respectively;the frequencies of the BCRP 421C ＞ A variant alleles were 31.97％ and 9.44％,respectively.And these differences were statistically significant (P ＜ 0.05).There was no significant difference in the distribution of allele frequencies between test group and Japanese (P ＞ 0.05).Conclusion Significant differences were observed in the distributions of some SNPs between Chinese and other populations.It is important to analyze the distribution of the related genetic polymorphisms of drug transporters in Chinese people to guide rational drug use in clinical.

Allele frequencies for 15 short tandem repeat (STR) loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) were obtained from 7,636 unrelated individuals of Chinese Han population living in Qinghai and Chongqing, China. Totally 206 alleles were observed, with the corresponding allele frequencies ranging from 0.0001-0.4982. Chi-square test showed that all of the STR loci agreed with the Hardy-Weinberg equilibrium. We also compared our data with previously published population data of other ethnics or areas. The results are valuable for human identification and paternity testing in Chinese Han population.

Breast cancer is one of the leading causes of cancer death worldwide. This study aimed to analyze the expression of centromere protein H (CENP-H) in breast cancer and to correlate it with clinicopathologic data, including patient survival. Using reverse transcription-polymerase chain reaction and Western blotting to detect the expression of CENP-H in normal mammary epithelial cells, immortalized mammary epithelial cell lines, and breast cancer cell lines, we observed that the mRNA and protein levels of CENP-H were higher in breast cancer cell lines and in immortalized mammary epithelial cells than in normal mammary epithelial cells. We next examined CENP-H expression in 307 paraffin-embedded archived samples of clinicopathologically characterized breast cancer using immunohistochemistry, and detected high CENP-H expression in 134 (43.6%) samples. Statistical analysis showed that CENP-H expression was related with clinical stage (P = 0.001), T classification (P = 0.032), N classification (P = 0.018), and Ki-67 (P < 0.001). Patients with high CENP-H expression had short overall survival. Multivariate analysis showed that CENP-H expression was an independent prognostic indicator for patient survival. Our results suggest that CENP-H protein is a valuable marker of breast cancer progression and prognosis.
Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Breast ; cytology ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line ; Cell Line, Tumor ; Chromosomal Proteins, Non-Histone ; genetics ; metabolism ; Epithelial Cells ; cytology ; metabolism ; Female ; Follow-Up Studies ; Humans ; Ki-67 Antigen ; metabolism ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Survival Rate ; Up-Regulation

BACKGROUNDInoperable chronic thromboembolic pulmonary hypertension (CTEPH) is a severe clinical syndrome characterized by right cardiac failure and possibly subsequent liver dysfunction. However, whether serum markers of liver dysfunction can predict prognosis in inoperable CTEPH patients has not been determined. Our study aimed to evaluate the potential role of liver function markers (such as serum levels of transaminase, bilirubin, and gamma-glutamyl transpeptidase [GGT]) combined with 6-min walk test in the prediction of prognosis in patients with inoperable CTEPH.

METHODSFrom June 2005 to May 2013, 77 consecutive patients with inoperable CTEPH without confounding co-morbidities were recruited for this prospective cohort study. Baseline clinical characteristics and 6-min walk distance (6MWD) results were collected. Serum biomarkers of liver function, including levels of aspartate aminotransferase, alanine aminotransferase, GGT, uric acid, and serum bilirubin, were also determined at enrollment. All-cause mortality was recorded during the follow-up period.

RESULTSDuring the follow-up, 22 patients (29%) died. Cox regression analyses demonstrated that increased serum concentration of total bilirubin (hazard ratio [HR] = 7.755, P < 0.001), elevated N-terminal of the prohormone brain natriuretic peptide (HR = 1.001, P = 0.001), decreased 6MWD (HR = 0.990, P < 0.001), increased central venous pressure (HR = 1.074, P = 0.040), and higher pulmonary vascular resistance (HR = 1.001, P = 0.018) were associated with an increased risk of mortality. Serum concentrations of total bilirubin (HR = 4.755, P = 0.007) and 6MWD (HR = 0.994, P = 0.017) were independent prognostic predictors for CTEPH patients. Patients with hyperbilirubinemia (≥23.7 &mu;mol/L) had markedly worse survival than those with normobilirubinemia.

CONCLUSIONElevated serum bilirubin and decreased 6MWD are potential predictors for poor prognosis in inoperable CTEPH.

Aged ; Antihypertensive Agents ; therapeutic use ; Bilirubin ; blood ; Exercise Test ; Female ; Humans ; Hypertension, Pulmonary ; blood ; drug therapy ; pathology ; Male ; Middle Aged ; Prognosis ; Prospective Studies

Objective To explore the effect of gene mutations of arsenic transport proteins-muhidrug resistance-associated proteins(MRP1 and MRP2)on phenotype of endemic arsenic poisoning.Methods Two hundreds and thirty-nine rural residents in 3 villages of Shuocheng Region,Shanxi Province were interviewed and examined by simple random sampling who had been lived there for 20 yearn at least.All the objects were divided into two groups on the basis of clinical examination with"The Standard Diagnosis of Endemic Arsenic Poisoning" (WT/S 211-2001):subjectives with skin lesion as a arsenic poisoning group and without skin lesion as a control group. One hundred and ninety-three blood samples were collected from each participanL Seventy-five arsenic poisoning cases and 118 controls were detected the gene mutations in the 2,17,23 exons of M RPI and the 10,18,31 exons of MRP2 by PCR-single strand conformation polymorphism (PCR-SSCP) and compared by multivariate Logistic regression model. Results Seventy-five cases and 164 controls underwent questionnaires. Age[ (58.85±11.26) vs (45.73±11.92),OR = 3.378,P ＜ 0.05],gender[male,57.3%(43/75)vs 27.4%(45/164),OR = 3.553,P＜ 0.01 ],smoking[46.7%(35/75) vs 21.3%(35/164),OR = 3.225,P ＜ 0.01 ],drinking[ 17.3%(13/75) vs 8.5% (14/164),OR = 1.836,P ＞ 0.05],vegetable and fruit intake[5.3%(4/75) vs 9.1%(15/164),OR = 0.560,P ＞ 0.05],egg and meat intake[34.7%(26/75) vs 30.5%(50/164),OR = 1.210,P ＞ 0.05],exposure of pesticide [41.3%(31/75) vs 29.3%(48/164),OR = 1.864,P ＜ 0.05] were tested by Logistic regression model. There was no gene mutation detected in the 23 exon of MRP1 and the 18 exon of MRP2. The gene mutations frequencies of the 2 exons of MRP1 in arsenic poisoning and control groups were 8.00% (6/75) and 5.93% (7/118),respectively;they were 13.33%(10/75) and 8.47%(10/118) of the 17 exons of MRP1,respectively;they were 22.67%(17/75) and 18.64%(22/118) of the 10 exons of MRP2,respectively;they were 5.33%(4/75) and 2.54%(3/118) of the 31 exons of MRP2,respectively. There was no significant difference between two groups(x2 = 0.312,1.165,0.460, 2.794,respectively,all P ＞ 0.05). After age,gender,smoking,drinking,nutritional level and exposure of pesticide being adjusted by multivariate Logistic regression model,there was no significant difference between two groups (OR = 0.803,1.892,2.388,1.098,respectively,all P ＞ 0.05). Conclusions The gene mutations of 2,17,23 exons of MRPI and the 10,18,31 exons of MRP2 may have no effect on the phenotype of endemic arsenic poisoning.

Objective To explore the mechanisms of different cholic acid for reducing damage to human liver cells lines L-O2 induced by amanita toxic peptides (amataxins).Methods According to different concentrations of amataxins,the experiment was conducted with different dosages in 5 groups:0.00 g/L,0.26 g/L,0.40 g/L,1.40 g/L and 2.80 g/L.The human liver cells lines L-O2 in the exponential growth phase were cultured into 96-well plates,1 ×103 cells per well After 24 hours,the concentrations of amanita toxic peptides mentioned above were added.The minimum concentration of mushroom toxins keeping the liver cells alive was determined after 24,48 and 72 hours,respectively,and MTT method was used to test each group's liver cell activity.The experiment included 3 groups:the control group,the damage group,and the cholic acid group.Each group had 3 time points:24,48 and 72 hours after being attacked.Twenty four hours after attack,in cholic acid group,cholic acid drugs including taurocholic acid gca,goose deoxycholic acid,gansu ammonia goose deoxycholic acid and bovine goose deoxycholic acid were given,respectively,to protect the injured liver cells.Cells' morphology changes were observed under the inverted phase contrast microscope,living cells were counted by using MTT method,and aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities in the culture supernatant were tested by the biochemical method.Results The minimum attack concentration of lamanita toxic peptides keeping liver cell survival in vitro was 1.40 g/L.Seventy-two hours after attack by amanita toxic peptides,the absorbance value was 0.812 ± 0.035,0.345 ± 0.021,0.363 ± 0.018,0.387 ± 0.027,0.431 ± 0.018,0.465 ± 0.015 and 0.452 ± 0.030,respectively in the control group,the damage group,the taurocholic acid group,the goose deoxycholic acid group,the glycocholic acid group,the glycochenodeoxycholic acid group and the sodium deoxycholic acid group.Compared with the damage group,absorbance value 72 hours after attack in each cholic acid group gradually increased,and compared with damage group,the differences were statistically significant among goose deoxycholic acid group,glycocholic acid group,glycochenodeoxycholic acid group and sodium deoxycholic acid group(P ＜ 0.05).AST and ALT activity in each cholic acid group declined,and that in glycochenodeoxycholic acid group was the lowest.Compared with the damage group,the difference was statistically significant (P ＜ 0.01).Conclusions Cholic acid can protect human liver cells from the damage induced by amanita toxic peptides.Such effect may be related to the fact that both amanita toxic peptides and cholic acid are the substrates of NTCP and OATP.

Objective To investigate whether brachial-ankle pulse wave velocity(baPWV),independently and jointly with the level of body mass index(BMI),affect the risk of incident diabetes with hypertension. Methods The effects of baPWV and BMI level on incident diabetes with hypertension were evaluated using Kaplan-Meier curves (log-rank test) and multi logistical regression models after adjustment for various confounding factors. Results A total of 2 222 participants were recruited in this study. Higher baPWV (Q4) was associated with a greater risk of incident diabetes with hypertension simultaneously compared with baPWV in the lower quartiles(Q1-Q3),with HR 4.16(95% CI 2.14,8.09)in the crude model,2.52(95% CI 1.10,5.78) in modelI,and 2.45(95% CI 1.05, 5.70)in modelⅡ.The highest risk of diabetes with hypertension simultaneously was in the group with high baPWV and BMI≥25 kg/m2,with HR 11.50(95% CI 4.5,29.9)in the crude model,4.20(95% CI 1.4,13.1)in modelI,and 4.30(95% CI 1.4,13.3) in modelⅡ. Conclusions The joint effect of baPWV and BMI level on incident diabetes with hypertension simultaneously is a better predictor than their independent effect. The risk of incident diabetes with hypertension markedly increases in the subjects with BMI≥25 kg/m2and higher baPWV.