OBJECTIVETo explore the expressions of trefoil factor 1 (TFF1) and trefoil factor 3 (TFF3) in prostate cancer (PCa) and prostate intraepithelial neoplasia (PIN) and their clinical significance.

METHODSUsing immunohistochemistry, we detected the expressions of TFF1 and TFF3 in the prostatic tissues of 89 cases of PCa, 50 cases of PIN, and 65 cases of benign prostate hyperplasia (BPH), and evaluated their clinical significance.

RESULTSThe positive rates of TFF1 and TFF3 expressions were 77. 53% and 48. 31% in PCa and 66.00% and 30.00% in PIN, significantly higher than 49.23% and 13. 85% in BPH (P <0. 05). The expression of TFF1 was not correlated with Gleason score (P >0. 05), while that of TFF3 was significantly higher in the PCa cases with Gleason score ≤7 than in those with Gleason score > 7 (70. 00% vs 42. 03%, P <0. 05). No significant correlation was observed between TFF1 and TFF3 expressions in PCa (P >0. 05).

CONCLUSIONThe expressions of TFF1 and TFF3 may contribute to the occurrence and progression of PCa, and therefore could be used as laboratory indexes in the diagnosis, differential diagnosis, and prognosis of PCa.

Disease Progression ; Humans ; Immunohistochemistry ; Male ; Peptides ; metabolism ; Prognosis ; Prostatic Hyperplasia ; metabolism ; Prostatic Intraepithelial Neoplasia ; metabolism ; Prostatic Neoplasms ; metabolism ; Trefoil Factor-1 ; Trefoil Factor-3 ; Tumor Suppressor Proteins ; metabolism

OBJECTIVETo clone human intestinal trefoil factor (hITF/hTFF3) gene into an eukaryotic expression vector for its expression in eukaryotic cells.

METHODSThe total RNA was extracted from normal human colon mucosa, and transcribed into cDNAs using RT-PCR. hTFF3 gene was amplified by PCR and ligated into pGEMT vector by TA cloning method. After sequencing, the hTFF3 gene was transfered into the eukaryotic expression vector pCMV5-myc. The recombinant vector was transfected into 293-T cells, and the expression of the recombinant protein was detected by Western blotting.

RESULTS AND CONCLUSIONhTFF3 gene was successfully cloned from normal human colon mucosa. The vector pCMV5-myc-hTFF3 was reconstructed, and in 293-T cells transfected with the vector, hTFF3 expression was detected by Western blotting.

Blotting, Western ; Cell Line ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; genetics ; Humans ; Intestinal Mucosa ; metabolism ; Peptides ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; methods ; Trefoil Factor-2 ; Trefoil Factor-3

OBJECTIVETo investigate the changes of the goblet cells in the intestine during the restitution process of the gut barrier after hemorrhagic shock.

METHODSForty-nine Sprague-Dawley rats with body weight of 250-300 g were divided into control group (n=7) and experimental group (n=42). Rats in the experimental group was further divided into 6 groups (n=7 each) according to different time point at 1, 3, 6, 12, 24, and 36 hours after hemorrhagic shock resuscitation. The specimens from ileum tissue were taken to observe the morphological chan ges of the intestinal mucosa. The number of goblet cells was determined by light microscope and/or electron microscope. The contents of trefoil factor family 3 (TFF3) of goblet cells were examined using GC-9A gas chromatographic instrument.

RESULTSAfter hemorrhagic shock, mucosal epithelial injury was obvious in the small intestine. Tissue restitution was found after 3 hours, and mostly established after 12 hours. Following tissue restitution,the denuded mucosal surface was covered intensively by goblet cells. The number of goblet cells on the intestinal mucosa was reduced significantly from 243+/- 13 at 1 h to 157+/- 9 at 24 h (r=- 0.910, P< 0.01), and returned to normal level at 36 h. In the experimental group, the content of TFF3 in the intestinal mucosa increased significantly at 12 hours, decreased, but was still higher at 24 hours (t=3.24, P< 0.05).

CONCLUSIONSThe goblet cells play a key role in the restitution of intestinal mucosa. High expression of TFF3 may facilitate the intestinal mucosal restitution in the early phase.

Animals ; Goblet Cells ; metabolism ; Ileum ; cytology ; Intestinal Mucosa ; cytology ; metabolism ; pathology ; Neuropeptides ; metabolism ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic ; metabolism ; Trefoil Factor-3

OBJECTIVE: To explore the effect on proliferation and invasion of human papillary thyroid carcinoma K1 cells by application of small hairpin RNA (shRNA) silencing TFF3 gene expression. METHOD: Using liposome transfection method, TFF3-shRNA targeting of TFF3 gene will be transient transfected to papillary thyroid carcinama K1 cells, inducing the corresponding gene silencing. The experiment set up blank control group (Con group), negative control group (ConNC group) and interference group (TFF3-shRNA group). The TFF3 protein and mRNA expression were evaluated by RT-PCR, Real time-PCR, immunocytochemistry and Western blot in K1 cells after TFF3-shRNA transfected. CCK-8 method and Scratch test were used to detect the change of proliferation ability and invasion ability respectively. RESULT: (1) The recombinant plasmid Ca # HSH018037-4-HIVmU6 carrying TFF3-shRNA transfected K1 cells successfully. (2) RT-PCR and Real time-PCR detected the expression of TFF3 mRNA, which was 0.38 ± 0.11 times as many as the blank control group (P < 0.01) after TFF3 gene silenced. But the negative control group was 1.082 times of blank control group (P > 0.05). (3) Western blot show that after TFF3 gene silence induced TFF3 protein expression levels have decreased 59.5% (P < 0.01), The difference was statistically significant compared with the blank control group. (4) Cell scratch detects K1 cell invasion ability. The invasion ability of K1 cells in interference group (TFF3-shRNA group) reduced. The scratch width significantly decreased 57.1% than blank control group (P < 0.01). (5) CCK-8 kit detect cell proliferation ability. K1 cells grow significantly slower in the interference group (TFF3-shRNA group) than the blank control group through the analysis of the growth curve (P < 0.01). In the interference group (TFF3-shRNA group) proliferation inhibition rate of K1 cells at 6 h, 12 h, 24 h and 36 h, 48 h are 16.6%, 26.6%, 33.6%, 33.8%, 35.0% respectively. Compared with negative control group, proliferation ability of K1 cell decreased significantly. CONCLUSION Silenced TFF3 gene can cause the degradation of mRNA, reduce the protein translation , and inhibit the invasion and proliferation ability of K1 cell.
Carcinoma ; genetics ; pathology ; Carcinoma, Papillary ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Peptides ; genetics ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Thyroid Cancer, Papillary ; Thyroid Neoplasms ; genetics ; pathology ; Transfection ; Trefoil Factor-3

OBJECTIVETo investigate the synergistic anti-inflammatory effect of Radix Platycodon in combination with herbs for cleaning-heat and detoxification and its mechanism for Fel-targeting.

METHODSForty Wistar rats were randomly divided into five groups (8 per group): the sham-operated group, model group, Radix Platycodon group, Flos Lonicera and Fructus Forsythia (LF) group, and Radix Platycodon, Flos Lonicera and Fructus Forsythia combination (PLF) group, using a random number table. A rat chronic obstructive pulmonary disease (COPD) model was established by passive smoking and intratracheal instillation of lipopolysaccharide (LPS). The treatments started from the 15th day of passive smoking for a total duration of 14 days. At the end of the treatment, changes in the following measurements were determined: lung histopathology, inflammatory cytokines including tumor necrosis factor α (TNF-α), transforming growth factor β (TGF-β) and interleukin IL-1β (IL-1β) in bronchoalveolar lavage fluid (BALF), and mRNA expression of endogenous active substance intestinal trefoil factor 3 (TFF3) in the lung tissue.

RESULTSLight microscopy showed that compared with the sham-operated group, rats in the COPD model group had disrupted alveolar structure, collapsed local alveoli, significantly widened or even fused alveolar septa, and massive infiltration of inflammatory cells in the alveolar wall and interstitium. In addition, significant bronchial epithelium hyperplasia, partially shed epithelia, and marked inflammatory cell infiltration in the bronchial wall and its surrounding tissues were noticed. Electron microscopy showed that rats in the model group had degeneration of alveolar type II epithelial cell; reduction, breakage or even loss of cell surface microvilli; swollen mitochondria with disappearing cristae and vacuole-like structure; and, increased secondary lysosomes in alveolar macrophages. The TNF-α, TGF-β and IL-1β levels and white blood cell (WBC) count in BALF were significantly increased (P < 0.01 or P < 0.05) and TFF3 mRNA expression in the lung tissue was significantly reduced (P < 0.01). After treatment, the pathological morphology of lung injury was less severe in all three treatment groups. In addition, TGF-β and IL-1β and WBC count in BALF were decreased (P < 0.01 or P < 0.05), and TFF3 mRNA expression in the lung tissue was significantly increased in the PLF group (P < 0.01). Compared with the LF group, the IL-1β in BALF was significantly decreased P < 0.05), and TFF3 mRNA expression was significantly increased (P < 0.05) in the PLF group.

CONCLUSIONSRadix Platycodon synergizes with herbs for cleaning-heat and detoxification in reducing inflammatory injury in a rat model of COPD. The synergistic anti-inflammatory effect is reflected in the improvement in pathological changes and in the reduction of IL-1β levels in BALF. The mechanism of such synergistic action may be related to its effect on maintaining the TFF3 mRNA expression and Fel-targeting function.

Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; cytology ; Disease Models, Animal ; Drug Synergism ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Immunohistochemistry ; Lung ; drug effects ; pathology ; Male ; Microscopy, Electron ; Neuropeptides ; genetics ; metabolism ; Phytotherapy ; methods ; Plant Preparations ; therapeutic use ; Platycodon ; Polymerase Chain Reaction ; methods ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; pathology ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Sensitivity and Specificity ; Trefoil Factor-3