Objective To assess the feasibility of a novel 99Tcm labelled polypeptide analogue for interleukin-11 receptor ( IL11R) imaging in a bone metastases model for prostate cancer. Methods A novel circular polypeptide analogue of IL11 ( c( CGRRAGGSC ) ) was indirectly labeled with 99Tcm and the product was named as 99Tcm-DTPA-IL11 RR. The labeling efficiency, radiochemical purity and stability of the product were measured with paper chromatography and high performance liquid chromatography (HPLC). The biodistribution of 99Tcm-DTPA-IL11RR was investigated in 28 ICR normal mice. The organ radioactivity was measured as percentage activity of injection dose per gram of tissue ( %ID/g). The models of bone metastases from prostate cancer were established at the tibias of BALB/c nude mice bearing human prostate cancer PC-3 cells. The tumor bearing ( n= 5 ) and standard closed fracture nude mice models underwent both 99Tcm-DTPA-IL11RR and 99Tcm-methylene diphosphonate (MDP) scintigraphy study. The images were acquired at 0.5, 1,2, 4, 6, 8, 24 h after intravenous injection of the tracers. The competitive inhibition imaging was perfomed in three tumor bearing mice. One-way variance analysis was used. Results The labeling efficiency was 90.7%. The radiochemical purity of 99Tcm-DTPA-IL11RR in normal saline solution was (99.57 ±0.09)%, (99.29 ±0.18)%, (98.95 ±0.78)%, (98.67 ±0.11)%, (96.53 ±0.91)%, (95.20±0.70)%, (88.38 ±0.22)% and (36.17±1.29)% at room temperature after0, 1,2, 3, 4, 6, 8 and 24 h, respectively. The tracer radiochemical purity in the blood of ICR mice remained over 90% at 37 C for 6 h. The labeling compounds were excreted mainly through kidney. The peak uptake of bone ( ( 1.910 ±0.109) %ID/g) and liver ( (0.366 ±0.030) %ID/g) was at 4 h after injection. In the tumor bearing mice, the uptake of spine marrow and large joints of extremities was mild. The highest uptake at tumor region was at 4 h and persistent at 6-8 h after injection. The tumor to non-tumor ratios (T/NT) were 1.17 ±0.17, 2.20 ±0.29, 3.20 ±0.15, 3.67 ±0.23, 13.61±0.85, 9.45 ±0.37 and 3.33 ±0.30 at 0.5,1, 2, 4, 6, 8 and 24 h, respectively (F=621.54, P＜0.05). In the standard closed fracture models,high uptake of 99Tcm-MDP was shown at the fracture site, with no increased 99Tcm-DTPA-IL11RR uptake noted. The tumor uptake was significantly depressed after a pre-injection of the unlabeled polypeptide analogue. Conclusions The synthesis of 99Tcm-DTPA-IL11RR is stable and the labeling efficiency is high. It may be a potential molecular probe in metastatic bone imaging for prostate cancer.

Objective To investigate the direct inhibitory effects of 153Sm- DTPA-c (Cys-Gly-Arg-Arg-Ala-Gly-Gly-Ser-Cys) NH2 ( 153 Sm-DTPA-c (CGRRAGGSC)) on human prostate cancer PC-3 cells. Methods 153Sm-DTPA-c(CGRRAGGSC) was synthesized by the reaction of 153SmCl3 with DTPA-c(CGRRAGGSC) using indirect synthesis method. PC-3 cells in vitro culture were divided into four study groups, groug A ( the control, with PBS only), group B with 1.5 mg/L c ( CGRRAGGSC), group C with 370 kBq 153 SmCl3 and group D with 370 kBq 153Sm-DTPA-c(CGRRAGGSC). PC-3 cell growth was assayed by 3-(4, 5-dimethylthiazol-2-yl ) -2, 5-diphenyltetrazolium bromide (MTT) method. Cell cycle and apoptosis were analyzed by flow cytometry. The expression changes of interleukin 11 (IL11 ) and IL11 receptor (IL1 1 R) in PC-3 cells were examined by Western Blot. One way analysis of variance (ANOVA) and paired-t test were applied for statistic analysis. Results The labeling yield of 153Sm-DTPA-c (CGRRAGGSC) was 85% and the radiochemical purity was 95.8%. The specific activity of 153Sm-DTPA-c(CGRRAGGSC) was 1.32 × 105 MBq/μmol. Significant inhibitory effects on the growth of PC-3 cells were found in both group C and D, with a time-dependent manner. However, no obvious inhibition was found either in group A or in group B. After 48 h,significant differences of sub-G1 peak area were found among groups, (0. 98 ± 0. 18)%, (0. 35 ±0. 10)%, (4.05 ±0.28)% and (13.38 ±0. 89)% for group A, B, C and D, respectively. Furthermore,sexpression of ILl 1R in group D was significantly lower than that in group B and C with absorbance values 0. 339 ~ 0.014, 0.338 ~ 0.019, 0.226 ~ 0. 015 for group B, C and D, respectively. Absorbance values in groups B and C were not significantly different after treatment, compared with those before treatment; however, there was difference between absorbance values after and before treatment in group D ( t = 0. 405,1. 163,135.989,P>0.05 >0.05, <0.05). Conchluion 153Sm-DTPA-c(CGRRAGGSC) can directly in hibit the cell growth and expression of human prostate cancer cells PC-3.

OBJECTIVETo observe the suppressive effect on the expression of pro-inflammatory cytokines in liver from brain dead (BD) rats through inhibition of p38 mitogen-activated protein kinase (MAPK) signaling pathway by SB203580.

METHODSA total of 30 male Wistar rats weighing from 180 to 200 g were randomly divided into 3 experimental groups: (1) BD group (n = 10): brain death was induced in rats; (2) BD+SB203580 group (n = 10): brain death was successfully induced and SB203580 (10 mg/kg) was given through dorsal vein of penis. After brain death artificial ventilation was maintained for 6 hours and only those with mean arterial blood pressure more than 80 mm Hg (1 mm Hg = 0.133 kPa) were accepted as BD donors. (3) Control group (n = 10): living healthy rats. The expressions of TNFalpha and IL-1beta mRNA in liver tissues were analyzed by RT-PCR and the protein expressions of TNFalpha, IL-1beta and phosphorylated p38MAPK were detected by Western blot.

RESULTSThe phosphorylated p38MAPK detected in the liver in BD group was significantly increased compared with the control group (q = 172.53, P < 0.01), and the expressions of TNFalpha and IL-1beta mRNA and proteins in liver were also significantly increased in BD group compared with the control group (q = 123.99, 135.35, 243.09 and 192.23, respectively, P < 0.01). The phosphorylated p38MAPK was decreased in BD+SB203580 group and significantly decreased compared with the BD group (q = 63.90, P is less than 0.05), but higher than that in control group (q = 108.63, P < 0.01). The expressions of TNFalpha and IL-1beta mRNA and proteins in liver were significantly decreased in BD+SB203580 group compared with the BD group (q = 55.11, 98.13, 61.03 and 50.85, respectively, P < 0.01), but higher than that in control group (q = 68.89, 37.22, 182.06 and 141.38, respectively, P < 0.01). SB203580 can suppress the expression of pro-inflammatory cytokines in the liver of brain dead rats through the inhibition of p38MAPK signaling pathway which may reduce the immunogenicity of donor livers.

Animals ; Brain Death ; metabolism ; Imidazoles ; pharmacology ; Inflammation ; Interleukin-1beta ; metabolism ; Liver ; drug effects ; metabolism ; pathology ; Male ; Pyridines ; pharmacology ; Rats ; Rats, Wistar ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

The synergistic antibacterial strategy of peroxidase mediated chemodynamic therapy(CDT)and photothermal therapy(PTT)has been proven to effectively resist bacteria.However,the antibacterial effect against Pseudomonas aeruginosa is severely limited due to the factors such as short lifespan of reactive oxygen species(ROS)(<200 ns)and limited diffusion distance(about 20?200 nm).In this study,glucopyranose functionalized MoO3-x(P-MoO3-x)nanoenzyme was successfully synthesized using a one-pot hydrothermal method.This nanoenzyme exhibited both peroxidase-like activity and a photothermal effect.The combined antibacterial performance and biological safety of P-MoO3-x was analyzed and verified.By utilizing specific interactions with glucopyranose and lectin,P-MoO3-x nanoenzyme could target the surface of Pseudomonas aeruginosa.This targeted approach effectively shortened the range of hydroxyl radicals,significantly enhancing the antibacterial effect against Pseudomonas aeruginosa.Under photothermal action,P-MoO3-x could reach the optimal reaction effect at 70℃.Even at low concentrations of hydrogen peroxide(50 μmol/L),it released more hydroxyl radicals.In vitro antibacterial analysis experiments demonstrated that the inactivation efficiency of the P-MoO3-x antibacterial system against Pseudomonas aeruginosa(106 CFU/mL)exceeded 99%.Furthermore,in vivo experiments confirmed the significant therapeutic effects of P-MoO3-x in treating methicillin-resistant Staphylococcus aureus(MRSA)infected wounds and promoting wound healing,without producing toxicity to cells.In conclusion,P-MoO3-x exhibited excellent antibacterial ability and good biocompatibility,making it a promising anti-infective nanoenzyme with broad application prospects.

This study was purposed to investigate the effect of rapamycin on proliferation, apoptosis, cell cycle progression and the regulation of chemokine receptor CXCR4 on RPMI8226 cells. Different concentrations of rapamycin were used to treat the multiple myeloma cell line RPMI8226 for different times. The proliferation of the cells was detected by MTT assay; the apoptosis rate and cell cycle were determined by flow cytometry (FCM); apoptosis of cells was observed by inverted microscopy; the cylin D1, CXCR4 and mTOR mRNA expressions were detected by RT-PCR or FQ-PCR after treating RPMI8226 cells with different concentrations of rapamycin. The results indicated that the rapamycin could inhibit the proliferation of RPMI8226 cells and induce their apoptosis. The cell cycle was arrested at the G(0)/G(1) phase. PCR results showed the down-regulation of mTOR, cyclin D1 and mTOR mRNA expressions after treating RPMI8226 cells with different concentrations of rapamycin for 24 hours. It is concluded that the rapamycin significantly inhibits the growth of RPMI8226 cells in a dose-and time-dependent mannes and induce cell apoptosis. Cell cycle arrests at the G(0)/G(1) phase, may be due to the down-regulation of the mTOR and cyclin D1 expressions. In additions, the down-regulation of CXCR4 mRNA expression is correlated with the reduction of adhesion between myeloma cells and stromal cells.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Humans ; Receptors, CXCR4 ; metabolism ; Sirolimus ; pharmacology

OBJECTIVETo investigate effect of oxidized low-density lipoprotein (ox-LDL) on memory CD8T cell subpopulation differentiation in mice with autoimmune diabetes.

METHODSCultured splenic CD8T cells from pre-diabetic NOD mice isolated with magnetic beads were treated with 30 µg/mL ox-LDL and 10 U/mL interleukin-2 (IL-2) for 24 h and the control cells were treated with IL-2 only. Flow cytometry was used to determine the percentage of splenic CD8IFN-γT cells, expressions of CD8, CD44 and CD62L on the T cells, and the activation of T cell factor-1 (TCF-1) and STAT-3. The CD127memory T cells were purified and transplanted into the pre-diabetic NOD mice via the tail vein, and the blood glucose was recorded weekly and survival time of the mice was monitored.

RESULTSTreatment with ox-LDL significantly reduced islet β cell-specific cytotoxic CD8T cells as compared with the control group [(0.7∓0.03)% vs (2.7∓0.14)%, P<0.01]. The percentage of effector memory CD8T cells (Tem) in the total memory CD8T cells was reduced [(10.3∓0.71)% vs (30.3∓1.36)%, P<0.01] and that of stem cell-like memory T cells was significantly increased [(72.3∓3.8)% vs (55.1∓2.61)%, P<0.05] following ox-LDL treatment, which also resulted in significantly decreased activation of TCF-1 [(14.5∓0.82)% vs (34.2∓1.23)%, P<0.01] and pSTAT-3 [(3.3∓0.12)% vs (22.1∓1.1)%, P<0.01]. Transplantation of ox-LDL-treated memory T cells in pre-diabetic NOD mice obviously inhibited the increase of blood glucose and prolonged the survival time of the mice (P<0.05).

CONCLUSIONOx-LDL decreases the activation of transcriptional factors TCF-1 and phosphorylation of STAT-3, inhibits the formation of effector memory CD8T cells with long-term cytotoxicity, but promote the generation of stem cell-like memory CD8T cells, which result in suppression of islet β cell-specific effector cytotoxic CD8T cell differentiation to lessen autoimmune injury to the islet β cells.

OBJECTIVETo investigate the changes of topoisomerase IIalpha (TOP2A) and HER2/neu genes in pancreatic ductal adenocarcinomas of Chinese patients, and to determine their roles during carcinogenesis and tumor progression.

METHODSExpressions of TOP2A and HER2/neu proteins were detected by using immunohistochemistry, while gene amplifications of TOP2A and HER2/neu were assessed by using multi-color fluorescence in situ hybridization (FISH). All the samples were of paraffin embedded and 10% formalin fixed tissue, including 26 cases of pancreatic ductal adenocarcinomas with adjacent non-neoplastic pancreatic tissues, 10 cases of chronic panreatitis, and 10 cases of normal pancreas. The correlation between TOP2A and HER2/neu gene status was analyzed.

RESULTSBy immunohistochemistry, the nuclear positive index of TOP2A in pancreatic ductal adenocarcinomas varied from 0.5% to 70%, and the positive rate of HER2/neu in pancreatic ductal adenocarcinomas was 46.2% (12/26). By FISH, 9/10 TOP2A amplified adenocarcinomas showed TOP2A and HER2/neu gene coamplification, while one case with HER2/neu gene amplification adenocarcinoma showed no TOP2A amplification. No expression of TOP2A, HER2/neu proteins and no amplification of TOP2A and HER2/neu gene were detected in adjacent non-neoplastic pancreatic tissues, chronic pancreatitis tissues and normal pancreas. No relationship was found between protein expression and gene amplification of TOP2A and HER2/neu (P > 0.05). TOP2A gene amplification was significantly correlated with HER2/neu gene amplification (P < 0.01).

CONCLUSIONSProtein expression of TOP2A and HER2/neu are not associated with the gene amplification. There is a significant correlation between TOP2A amplification and HER2/neu gene amplification. Co-amplification of TOP2A and HER2/neu may play an important role in the carcinogenesis and progression of pancreatic carcinoma. Evaluation of the status of TOP2A and HER2/neu may be helpful to achieve target therapy of pancreatic carcinoma.

Adenocarcinoma ; genetics ; metabolism ; pathology ; secondary ; Adult ; Aged ; Antigens, Neoplasm ; genetics ; metabolism ; Carcinoma, Pancreatic Ductal ; genetics ; metabolism ; pathology ; secondary ; DNA Topoisomerases, Type II ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Gene Amplification ; Gene Expression Regulation, Neoplastic ; Genes, erbB-2 ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Liver Neoplasms ; metabolism ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Pancreatic Neoplasms ; genetics ; metabolism ; pathology ; Poly-ADP-Ribose Binding Proteins ; Receptor, ErbB-2 ; genetics ; metabolism

OBJECTIVETo investigate 18q21 LOH in human pancreatic ductal adenocarcinomas and chronic pancreatitis by fluorescence in-situ hybrydization (FISH) technique, and to analyze the relationship between 18q21 LOH and clinicopathologic characteristics.

METHODSRP11-729G3 and RP11-850A17, the regions on 18q21, were selected as the target fragments, the region RP11-621L6, close to the centromere of chromosome 18, was selected as the reference fragment. The specific BAC clones were used to isolate and purify the corresponding genomic DNA, which were labeled with biotin or DIG by nick translation into dual color probes. 18q21 LOH was assessed by dual-color FISH in 30 cases of pancreatic ductal adenocarcinoma and 10 cases of chronic pancreatitis. All samples were 10% formalin fixed and paraffin embedded. The relationship between 18q21 LOH and clinicopathologic characteristics was analyzed.

RESULTSAmong 30 cases of pancreatic ductal adenocarcinoma, 25 cases showed LOH at the region RP11-729G3 (83.3%), and 26 cases showed LOH at the region RP11-850A17 (86.6%). Among these, 25 cases with LOH at both regions, 1 case showed LOH only at the region of RP11-850A17. No LOH was found in 10 cases of chronic pancreatitis.

CONCLUSIONS18q21 LOH is a high-frequency event in human pancreatic ductal adenocarcinomas. LOH at the regions RP11-729G3 and RP11-850A17 demonstrates a high concordance. 18q21 may play an important role during pancreatic carcinogenesis and tumor progression. 18q21 LOH may be used as a diagnostic marker for pancreatic ductal adenocarcinoma.

Adenocarcinoma ; classification ; genetics ; Adult ; Aged ; Carcinoma, Pancreatic Ductal ; classification ; genetics ; Chromosome Mapping ; Chromosomes, Human, Pair 18 ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Loss of Heterozygosity ; genetics ; Male ; Middle Aged ; Pancreatic Neoplasms ; classification ; genetics ; Pancreatitis, Chronic ; classification ; genetics

Echinostoma hortense (Digenea: Echinostomatidae) is one of the intestinal flukes with medical importance in humans. However, the mitochondrial (mt) genome of this fluke has not been known yet. The present study has determined the complete mt genome sequences of E. hortense and assessed the phylogenetic relationships with other digenean species for which the complete mt genome sequences are available in GenBank using concatenated amino acid sequences inferred from 12 protein-coding genes. The mt genome of E. hortense contained 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and 1 non-coding region. The length of the mt genome of E. hortense was 14,994 bp, which was somewhat smaller than those of other trematode species. Phylogenetic analyses based on concatenated nucleotide sequence datasets for all 12 protein-coding genes using maximum parsimony (MP) method showed that E. hortense and Hypoderaeum conoideum gathered together, and they were closer to each other than to Fasciolidae and other echinostomatid trematodes. The availability of the complete mt genome sequences of E. hortense provides important genetic markers for diagnostics, population genetics, and evolutionary studies of digeneans.
Amino Acid Sequence ; Base Sequence ; Databases, Nucleic Acid ; Dataset ; Echinostoma* ; Echinostomatidae ; Fasciolidae ; Genes, rRNA ; Genetic Markers ; Genetics, Population ; Genome ; Genome, Mitochondrial* ; Humans ; RNA, Transfer ; Trematoda

Objective:To assess the deformation of the bladder-neck opening and the impact of the bladder-neck angle(BNA)on urination in male patients by fluid-structure interaction(FSI)analysis.Methods:We established geometric models of the blad-der,prostate and urethra were established,incorporating both normal and enlarged BNAs,and assessed the effects of BNA alteration on urinary flow by FSI simulation of the flow rate and pressure of the urine within the bladder,bladder neck and urethra,and that of pros-tate displacement as well.We retrospectively analyzed the clinical data on 145 male patients from the Second Hospital of Hebei Medical University between June 2020 and June 2023,39 with acute urine retention(the AUR group)and 106 without(the non-AUR group),and evaluate the impact of BNA on urination based on the urinary flow rate and prostate volume.Results:Comparative simulation a-nalysis showed significant differences in the total urethral pressure and flow rate between the normal and enlarged BNA models(P＜0.05).The maximum prostate displacement was found at the bladder neck,with moderate displacement and unchanged urethral diame-ter in the normal BNA model,but significant displacement and a reduced urethral opening diameter in the enlarged BNA model.FSI analysis confirmed an evident impact of enlarged BNA on urination,more significant in the AUR than in the non-AUR patients(P＜0.05).The BNAs in the patients with the maximum urinary flow rate(Qmax)of＜10,10-15 or＞15 ml/s were 83.7°±2.5°,67.5°±1.8° and 65.1°±4.8° respectively,with statistically significant difference between the former one and the latter two groups(P＜0.05).The BNAs in the patients with normal prostate volume or BPH of grade Ⅰ,Ⅱ,Ⅲ or Ⅳ were 65.0°±3.7°,67.2°±3.1°,71.5°±2.0°,82.8°±3.5° and 105.8°±6.0°,respectively(P＜0.05),with statistically significant difference between BPH grades Ⅲ and Ⅳ(P＜0.05)as well as between these two and the other three groups(P＜0.05),but not among the normal prostate volume,BPH grade Ⅰ and BPH grade Ⅱ groups(P＞0.05).Spearman correlation analysis indicated that BNA was strongly correlated with total prostate volume(TPV),transition zone volume(TZV),intravesical prostatic protrusion(IPP),prostatic urethral angle(PUA),IPSS,and Qmax(P＜0.05).Conclusion:Changes in BNA affect urination and are closely associated with the se-verity of prostate hyperplasia.The BNA may be an important anatomical factor for assessing the severity of lower urinary tract symptoms in BPH patients.