Pestiviruses infection: Interferon-virus mutual regulation.

Pestiviruses are a class of viruses that in some cases can cause persistent infection of the host, thus posing a threat to the livestock industry. Interferons (IFNs) are a group of secreted proteins that play a crucial role in antiviral defense. In this review, on the one hand, we elaborate on how pestiviruses are recognized by the host retinoic acid-inducible gene-I (RIG-I), melanoma-differentiation-associated protein 5 (MDA5), and Toll-like receptor 3 (TLR3) proteins to induce the synthesis of IFNs. On the other hand, we focus on reviewing how pestiviruses antagonize the production of IFNs utilizing various strategies mediated by self-encoded proteins, such as the structural envelope protein (Erns) and non-structural protein (Npro). Hence, the IFN signal transduction pathway induced by pestiviruses infection and the process of pestiviruses blockade on the production of IFNs intertwines into an intricate regulatory network. By reviewing the interaction between IFN and pestiviruses (based on studies on BVDV and CSFV), we expect to provide a theoretical basis and reference for a better understanding of the mechanisms of induction and evasion of the innate immune response during infection with these viruses.

[Expression of Ras-Associated Protein 1 in the Vascular Endothelium of Bone Tissue with Non-Traumatic Osteonecrosis of Femoral Head].

Objective: To investigate the difference in the expression of Ras-associated protein 1 (Rap1) in necrotic and healthy areas of non-traumatic osteonecrosis of femoral head (NONFH) patients. Methods: Femoral head tissue samples from 30 cases of NONFH and 30 cases of traumatic osteonecrosis of the femoral head (TONFH) were collected after hip replacement surgery, respectively. No significant difference of Association Research Circulation Osseous (ARCO) staging was found between the NONFH and the TONFH groups ( Z=-0.769, P=0.442). In the NONFH group, 8 patients were ARCO stage IIIb, 10 were stage IV, and 12 were stage V, while in the TONFH ground, 11 patients were ARCO stage IIIb, 9 were stage IV, and 10 were stage V. There were 19 males and 11 females in the NONFH group, with an average age of 49.6 yr. (26-69 yr.), and 16 males and 14 females in the TONFH group, with an average age of 54.2 yr. (37-68 yr.). There was no significant difference in gender or age between the two groups ( P>0.05). Specimens were collected from different bone areas, including those from the necrotic areas (area A) and the healthy areas (area B) of the NONFH group, and those from the healthy areas (area B') of the TONFH group, i.e., the control group. Western blot and quantitative real-time reverse transcription PCR (qRT-PCR) were used to analyze the different expression of Rap1, vascular endothelial growth factor (VEGF) protein, phosphoinositide 3-kinase (PI3K), and Akt protein and their corresponding mRNA in the three areas of bone tissue. HE staining and immunohistochemisty staining were done in order to observe the morphological changes of each area. Results: Western blot results indicated that there was no statistical difference in the relative expression of Rap1, VEGF, PI3K, and Akt proteins ( P>0.05). The relative expressions of Rap1, VEGF, PI3K, and Akt proteins in the area A were lower than those in the area B and the difference was statistically significant ( P<0.05). qRT-PCR results showed that the relative expressions of Rap1, VEGF, PI3 K and Akt mRNA in area A were lower than those of area B, and a statistical difference was found ( P<0.05). The relative expression of the mRNA of Rap1, VEGF , PI3 K and Akt in area B and area B' were not significantly different ( P>0.05). HE staining and immunohistochemisty staining showed that chondrocytes decreased in the necrotic area (area A) of NONFH, chondrocytes nucleus disappeared, subchondral bone trabeculae were broken, bone trabeculae thickened, and empty bone lacunae appeared. Granulation tissues composed of new capillaries and fibrous cells have proliferated and crawled around the necrotic area. Positive expressions of the Rap1, VEGF, PI3K and Akt proteins in area A were weaker than those of the normal area. In addition, there were positive expressions of Rap1, PI3K and Akt on the trabecular bone of both area A and area B at similar intensity of expression. There were strong positive expressions of Rap1, VEGF, PI3K and Akt on the intima of arterioles and venules, and on the peripheral stromal cell membrane, but the positive expression in area A was significantly lower than that in area B. However, the positive expression positions and intensity of all indicators were similar in area B and area B'. Conclusion: The necrosis in NONFH may be related to vascular endothelial damages caused by the inhibition of the Rap1-PI3K/Akt signaling pathways and the subsequent decline in the protein expression.

T Follicular Helper Cells Regulate Humoral Response for Host Protection against Intestinal Citrobacter rodentium Infection.

Citrobacter rodentium colonizes at the colon and causes mucosal inflammation in mice. Previous studies have revealed the importance of the innate and adaptive immune response for controlling C. rodentium infection. In the present study, we examined the role of T follicular helper (Tfh) cells in intestinal C. rodentium infection using mice with Bcl6 deficiency in T cells. Tfh cells were absolutely required at the late, but not the early, phase to control infection. Compared with control mice, we observed systemic pathogen dissemination and more severe colitis in Tfh-deficient mice. Furthermore, the susceptibility of Tfh-deficient mice correlated with an impaired serum IgG1 response to infection, and serum Abs from infected wild-type mice protected Tfh-deficient mice from infection. The transfer of wild-type Tfh cells also restored the levels of IgG1 and led to effective clearance of the pathogens in Tfh-deficient mice. Moreover, during C. rodentium infection, IL-21- and IL-4-producing Tfh cells were increased obviously in wild-type mice, correlating with IgG1 as the major isotype in germinal center B cells. Taken together, our work highlights the requirement and the function of Tfh cells in regulating humoral response for the host protection against C. rodentium infection.

Up-regulation of TIMP-3 and RECK decrease the invasion and metastasis ability of colon cancer.

BACKGROUND AND STUDY AIMS: Although the function of microRNA21 (miR-21) in the invasion and metastasis of colon cancer has been extensively studied, the mechanisms of invasion and migration related pathways between and its targets are still not elucidated. This study explored the mechanisms of the pathway between miR-21 and the target genes in vitro and in vivo. MATERIALS AND METHODS: We transfected pmiRZip21 or Leti3 into colon cancer cells. The levels of miR-21 expression, mRNA transcription and protein of target genes were analysed by TaqMan microRNA assays, RT-PCR and western blot, respectively. Scratch migration and trans-well assays were used to evaluate metastasis and invasion. To build a subcutaneous tumour animal model, detect the level of miR-21 and the target genes and then identify the mechanisms in vivo. RESULTS: MiR-21 expression levels in colon cancer cells transfected with pmiRZip21 in vivo or in vitro were decreased (P < 0.05). The mRNA and protein levels of TIMP-3 and RECK were up-regulated after inhibiting miR-21 in vitro and in vivo (P < 0.05), but those of BMPR-II and PCDH17 were not. In pmiRZip21-transfected colon cancer cells, invasion and migration were significantly decreased both in vitro and vivo (P < 0.05). CONCLUSIONS: Up-regulation of TIMP-3 and RECK, by inhibiting miR-21 expression can decrease tumour invasion and metastasis ability in vitro and in vivo, and has potential as a possible target site in anti-tumour therapy. More effects in vivo have to be investigated in further research.

Sensitive FRET Biosensor Reveals Fyn Kinase Regulation by Submembrane Localization.

Fyn kinase plays crucial roles in hematology and T cell signaling; however, there are currently limited tools to visualize the dynamic Fyn activity in live cells. Here we developed and characterized a highly sensitive Fyn biosensor based on fluorescence resonance energy transfer (FRET) to monitor Fyn kinase activity in live cells. Our results show that Fyn kinase activity can be induced in both mouse embryonic fibroblasts (MEFs) and T cells by ligand engagement. Two different motifs were further introduced to target the biosensor at the cellular membrane microdomains in MEFs, revealing that the Fyn-tagged biosensor had 70% greater response to growth factor stimulation than the Lyn-tagged version. This suggests that the plasma membrane microdomains can be categorized into different functional subdomains. Further experiments show that while the membrane accessibility is necessary for Fyn activation, the localization of Fyn outside of its microdomains causes its hyperactivity, indicating that membrane microdomains provide a suppressive microenvironment for Fyn regulation in MEFs. Interestingly, a relatively high Fyn activity can be observed at perinuclear regions, further supporting the notion that the membrane microenvironment has a significant impact on the local molecular functions. Our work hence highlights a novel Fyn FRET biosensor for live cell imaging and its application in revealing an intricate submembrane regulation of Fyn in live MEFs.

Engineered proteins with sensing and activating modules for automated reprogramming of cellular functions.

Protein-based biosensors or activators have been engineered to visualize molecular signals or manipulate cellular functions. Here we integrate these two functionalities into one protein molecule, an integrated sensing and activating protein (iSNAP). A prototype that can detect tyrosine phosphorylation and immediately activate auto-inhibited Shp2 phosphatase, Shp2-iSNAP, is designed through modular assembly. When Shp2-iSNAP is fused to the SIRPα receptor which typically transduces anti-phagocytic signals from the 'don't eat me' CD47 ligand through negative Shp1 signaling, the engineered macrophages not only allow visualization of SIRPα phosphorylation upon CD47 engagement but also rewire the CD47-SIRPα axis into the positive Shp2 signaling, which enhances phagocytosis of opsonized tumor cells. A second SIRPα Syk-iSNAP with redesigned sensor and activator modules can likewise rewire the CD47-SIRPα axis to the pro-phagocytic Syk kinase activation. Thus, our approach can be extended to execute a broad range of sensing and automated reprogramming actions for directed therapeutics.Protein-based biosensors have been engineered to interrogate cellular signaling and manipulate function. Here the authors demonstrate iSNAP, a tool to detect tyrosine phosphorylation and activate desired protein enzymes allowing the control of phagocytosis in macrophages.

Dendritic cells transfected with indoleamine 2,3-dioxygenase gene suppressed acute rejection of cardiac allograft.

BACKGROUND: Immunomodulation by indoleamine 2,3-dioxygenase (IDO) has been documented in many studies yet its underlying mechanisms remain undefined, especially in solid organ transplantation. Recent research demonstrated that the active expression of IDO in dendritic cells (DCs) regulates immune reaction. This study assessed whether DCs transfected with IDO gene inhibit T cells responses and suppress cardiac allograft rejection. METHODS: Adenovirus vector containing IDO gene was transfected into DCs to obtain IDO-positive DCs (IDO(+) DCs). To evaluate the effect of IDO(+) DCs on T cells in vitro, CD4(+) T cell proliferation and apoptosis was assessed in mixed lymphocyte reactions and measured by flow cytometry, respectively. IDO(+) DCs from C57BL/6 mice were injected into BALB/c recipients before heterotopic cardiac transplantation. RESULTS: Supernatant fluids from cultures of IDO(+) DCs had decreased tryptophan and increased kynurenine levels, reflecting IDO activity. IDO(+) DCs suppressed CD4(+) T cell responses in vitro, as reflected by decreased proliferation and increased apoptosis. In the transplant model, IDO(+) DCs prolonged survival and alleviated rejection of cardiac allograft in recipients injected with IDO(+) DCs. In vivo, IDO(+) DCs also significantly impaired CD4(+) T cell responses promoting increased apoptosis and a Th2-dominant cytokine shift. CONCLUSIONS: IDO overexpression in DCs suppressed T cells alloresponses in vitro, and IDO(+) DCs attenuated acute allograft rejection in vivo. Regulation of tryptophan catabolism by means of IDO overexpression in DCs may be a useful approach in cardiac transplantation and immunological tolerance.

[Location of bone marrow mesenchymal stem cells in rats after small intestinal transplantation].

OBJECTIVE: To study the bone marrow mesenchymal stem cells (MSC) settled in rats after small intestinal transplantation. METHODS: Bone marrow MSCs were taken from 1-month male Lewis rats, isolated and cultured by density gradient centrifugation and differential adherent culture. The surface antigens (CD29, CD90, CD34 and CD45) of MSC were identified by flow cytometry. Final concentration of 5 µg/L CFSE was used to mark the third generation of MSCs. Adult male inbred line F344 rats were used as donor and adult male Lewis rats as acceptor. A heterotopic intestinal transplant rat model was established by F344 to Lewis. Labeled MSCs were injected into model rats through vena dorsalis penis after operation. Tissues at postoperative 7-day were collected for frozen pathology to reveal the location of transplanted MSCs under fluorescence microscope. RESULTS: MSCs were successfully isolated from rat bone marrow. The average positive expression rates of surface antigens CD29, CD90, CD34 and CD45 were 96.48%, 99.77%, 2.41% and 1.39% respectively. MSCs were successfully and effectively marked with CFSE. Seven days after operation, a large number of green fluorescence could be observed in transplanted intestine, spleen and thymus. Autograft intestinal tissues only showed trace fluorescence, and the heart, liver and lung tissue basically did not present the green fluorescence. CONCLUSION: Bone marrow MSCs can settle in transplanted small intestine of rat.

[Research on the suppressive effect on transplantation rejection by indoleamine 2, 3-dioxygenase].

OBJECTIVE: To study the suppressive effect of indoleamine 2, 3-dioxygenase on transplantation rejection in mice heterotopic cardiac transplantation. METHODS: Adenovirus vector containing IDO gene was used to infect donor (C57BL/6) DC to obtain IDO(+)DC. Mouse heterotopic cardiac transplantation models were established (C57BL/6-BALB/c) and the following groups were set up, including the control group, DC injection group, TC injection group, IDO(+)DC injection group and co-injection group of IDO(+)DC and TC, 12 donors and 12 recipients in each group.Survival time of the donor heart in every group was observed. Meanwhile, donor hearts were harvested 7 days post transplantation for different examinations, including pathological examination, mRNA expression of IDO through real-time PCR, IDO protein expression through Western blot. Peripheral blood of recipients was also harvested for CD3(+)T lymphocyte apoptosis rate examination through fluorescence-activated cell sorting.One-way ANOVA and Kaplan-Meier Survival Analysis were used for statistic analysis of IDO expression, CD3(+)T lymphocyte apoptosis rate and survival time of the donor heart respectively. RESULTS: Cadiac allograft median survival time of each group were 7.0, 7.5, 11.0, 17.5, 24.0 days respectively. Compared with control and DC injection group, IDO(+)DC, TC and co-injection group significantly prolonged the survival time of donor hearts (t = 3.523-8.449, P < 0.01). Both IDO mRNA and protein expression showed significant increase(t = 5.974-16.176, P < 0.01). The CD3(+)T lymphocyte apoptosis rate was also significantly increased (t = 6.324-38.120, P < 0.01). Compared with IDO(+)DC or TC group alone, co-injection group significantly prolonged the survival time of the donor heart (t = 5.971 and 2.831, P < 0.05). Both IDO mRNA and protein expression showed significant increase (t = 2.853-15.194, P < 0.01).Furthermore, the CD3(+)T lymphocyte apoptosis rate was significantly increased as well (t = 26.069 and 7.643, P < 0.05). CONCLUSIONS: Suppressive effect of co-injection of IDO(+)DC and TC is much more effective than administration of IDO(+)DC or TC alone, which suggests that IDO achieved immune suppressive effect through the pathway of tryptophan depletion and accumulation of TC.

Emergency caudate lobectomy for ruptured hepatocellular carcinoma with multiple primary cancers.

We report a case of metachronous multiple primary malignancies involving both rectum and liver with colonic metastasis from hepatocellular carcinoma (HCC) through hematogenous pathway. A 72-year-old woman was admitted to the emergency department with right upper abdominal pain for 4 h. Considering her surgical history of Mile's procedure plus liver resection for rectal cancer with liver metastasis three years ago and the finding of urgent computed tomography scan on admission, the preoperative diagnosis was spontaneous rupture of rectal liver metastasis located in caudate lobe and colonic metastasis from rectal cancer. The patient underwent an emergency isolated caudate lobectomy at a hemorrhagic shock status. Pathology reported a primary HCC in the caudate lobe and colonic metastasis of HCC with tumor embolus in the surrounding vessels of the intestine. No regional lymph node involvement was found. It is hypothesized that HCC may disseminate hematogenously to the ascending colon, thus making it a rare case.

A novel method for detection of mutation in epidermal growth factor receptor.

BACKGROUND: For the rapid and sensitive screening of epidermal growth factor receptor (EGFR) hot-spot mutations, we developed a novel method combining mutant-enriched PCR with amplification refractory mutation system (ARMS) TaqMan real-time PCR in a one-step reaction tube. METHODS AND RESULTS: We designed two pairs of primers to enrich and genotyping each mutation (E746_A750del and L858R): nest primers and ARMS primers. Before the PCR assays were carried out, the restriction enzymes were used to cut wild alleles. The results showed that this method could detect mutant alleles mixed samples containing 0.1% with a cutoff ΔCt value of 12. We used this method in a survey of 73 non-small cell lung cancer (NSCLC) samples, detecting 14 mutant samples of E746_A750del and 12 mutant samples of L858R. The results well agreed with the results of DxS. All unmatched samples were identified by sequencing and the results showed that our method has high specificity. CONCLUSION: The mutant-enriched ARMS TaqMan PCR could be useful in the detection of mutation in clinical samples containing only a small number of mutant alleles.

[Enteral versus parenteral nutrition after gastrointestinal surgery: a meta-analysis of randomized controlled trials].

OBJECTIVE: To compare the different prognosis between enteral nutrition (EN) and parenteral nutrition (PN) in patients after gastrointestinal surgery (GIS), and to investigate a reasonable regimen of enteral nutrition (EN) after GIS. METHODS: Randomized controlled trials (RCTs) on EN/PN after GIS from 1970 to 2008 retrieved from the data bank of Pubmed, EMBASE and Cochrane Library were analyzed. Evaluation endpoints were anastomotic dehiscence, infection (catheter sepsis, wound infection, pneumonia, intra-abdominal abscess and urinary tract infection), vomiting and abdominal distention, other complications, length of hospital stay and mortality rate. RESULTS: Twenty-three RCTs including 2784 patients met the entering criteria. Compared with PN, EN was beneficial in the reduction of anastomotic dehiscence (RR = 0.67, 95%CI: 0.50 - 0.91; P = 0.010), infections (RR = 0.72, 95% CI: 0.64 - 0.81; P < 0.001), other complication (RR = 0.82, 95%CI: 0.73 - 0.92; P < 0.001) and duration of hospital stay (weighted mean difference: -3.60; 95%CI: -3.88 - -3.32; P < 0.001). But the risk of vomiting was increased among patients with EN (RR = 1.39, 95%CI: 1.21 - 1.59; P < 0.001), and there was no significant differences in mortalities between the two groups (P = 0.400). CONCLUSIONS: There is no advantage in treating patients 'nil by mouth' after gastrointestinal surgery. It indicated that early commencement of enteral feeding is beneficial.

[Efficacy and safety of adjuvant post-surgical therapy with imatinib in gastrointestinal stromal tumor patients with high risk of recurrence: interim analysis from a multicenter prospective clinical trial].

OBJECTIVE: To evaluate the efficacy and safety of postoperative adjuvant chemotherapy with imatinib in gastrointestinal stromal tumor(GIST) patients who had high risk of recurrence. METHODS: A prospective, open-label, multi-center trial conducted in sixteen teaching hospitals in China was carried out. The criteria of the enrolled patients included age more than 18 years old, CD117 positive GIST, tumor size more than 5 cm, pathological mitosis counts more than 5/50 HPF, and treatment beginning within 4 weeks after complete resection and with imatinib (400 mg, once a day) for at least 12 months. The 1, 3 year recurrence rates, disease free survival, overall survival rate and quality of life were evaluated. RESULTS: From Aug. 16th 2004 to Sep. 13th 2005, there were totally 74 patients screened and 57 patients (34 men, 23 women) enrolled in the imatinib treatment group. The primary tumors were located in the stomach in 50.9%, the small intestine in 38.6% and the colorectum in 10.5% of the cases. All the patients received radical resection. Until the cut-off date of interim analysis, there was no evidence of tumor relapse or metastasis in all patients and no death was reported either. Among the 57 enrolled patients with intention to treat(ITT), twelve patients finished the protocol (per protocol, PP). The disease free survival was (268.3 +/-120.2) d in ITT analysis, and (396.7+/-38.2) d in the PP analysis. The incidence of adverse effect was 44.4% . The score in quality of life showed no statistically significant difference between the baseline visit and the follow-up visits. CONCLUSION: Imatinib is a promising postoperative adjuvant chemotherapy in GISTs patients with high risk of recurrence, and the adverse effects are receivable.

[Effect of 5-fluorouracil in combination with Astragalus membranaceus on amino acid metabolism in mice model of gastric carcinoma].

OBJECTIVE: To study the effect of 5-fluorouracil-FU in combination with astragalus membranaceus(AM) on amino acid metabolism in mice model of gastric carcinoma induced by 3-methylcholanthrene(MC). METHODS: Mice gastric carcinoma models were established by 3-methylcholanthrene induction and randomly divided into different groups, and received 5-FU treatment (group A) 5-FU plus AM (group B), 5-FU plus a high dose of AM(group C), no treatment (group D). Normal mice were used as control (group N). Free amino acid in the tumor specimens were examined. RESULTS: The levels of free Valine, Methionine, Leucine, Arginine and cystine in the tumor specimens in group D were significantly higher than that in group N(P< 0.05). The levels of free serine in group A, B, C, D were significantly higher than that in group N. The levels of free glutamic acid in group A, B were significantly higher than that in group N(P< 0.05). The levels of free proline in group C, D were significantly higher than that in group P, N(P< 0.05). CONCLUSIONS: The increasing levels of free serine and proline in tumor specimens in gastric cancer mice model reveals metabolic disturbance of amino acid. 5-FU plus astragalus membranaceus can decrease the level of free glutamic acid in the mice models, and inhibit tumor growth.

[Clinical value of the sixth edition TNM stages analysing prognosis of colorectal cancer].

OBJECTIVES: To study the clinical value analyzing of colorectal cancer prognosis by the Sixth Edition TNM Stages. METHODS: 5481 cases with colorectal cancer and treated by operational methods, were collected. All the cases were separately staged by the Fifth Edition or Sixth Edition TNM Stages standards. The 5-year survival rates were analyzed by the life table method. RESULTS: The 5-year survival rates of the Fifth Edition TNM Stages of I, II, III and IV were 80.1%, 68.0%, 40.5% and 9.8% respectively. The 5-year survival rates of the Sixth Edition TNM Stages of II(A) and II(B) were 71.6% and 66.4% respectively, and of the stages III(A), III(B) and III(C) were 46.2%, 40.1% and 28.3% respectively. There were statistical differences among the sub-stages II and III, P < 0.05. CONCLUSION: The Sixth edition TNM Stages laid more stress on effect of the local infiltration depths and lymphatic metastasis in the prognosis of colorectal cancer, therefore, the stages were more fine, to analyze prognosis of the colorectal cancer were more precise. It is high clinical value for the individual complex treatment with every sub-stages.

[Diagnosis and surgical treatment of 48 cases of parathyroid adenoma and parathyroid carcinoma].

OBJECTIVE: To summarize the experience in diagnosis and surgical treatment of parathyroid adenoma and carcinoma (PTA and PTC) in our department. METHODS: The clinical and pathological data of 48 cases admitted in our department from Jan 1995 to Dec 2005 were reviewed. Among the 48 cases, 46 cases were of parathyroid adenoma and 2 cases of parathyroid carcinoma. The average clinical history of the 48 cases was 3.65 +/- 2.83 years. The serum calcium and PTH levels were elevated in all the 48 cases. In 31 cases ultrasonographic results were consisted with that of 99mTc-MIBI scintigraphy. Unilateral neck exploration was performed in 18 cases and no case with post-operative tumor remnants was found. In other 13 cases bilateral exploration was performed but no one case was found to be tumor positive in the opposite side of the glands. Tumors resection was performed in all the 48 cases, among which in the 2 cases with PTC, ipsilateral thyroid lobe excision and modified neck dissection were also performed. RESULTS: Clinical symptoms of all the patients were relieved after operation. No recurrent case was found during the follow-up periods (from 1 month to 10 years). The average level of serum calcium and PTH declined significantly after operation. The post-operational serum calcium and PTH levels at 3 days after operation were even lower than normal. Transient post-operational hypocalcemia was found in almost all the patients. The serum calcium and PTH levels in all patients recovered to normal level within a periods from 1 week to 3 months after operation. The sensitivity and positive prediction value of localization methods were 97.0% and 94.1% of ultrasonography, respectively, and 100% and 97.3% of 99mTc-MIBI scintigraphy, respectively. CONCLUSION: Patients with chronic bone diseases, repeatedly recurrent nephrolithiasis, peptic ulcer disease or pancreatitis should be regarded as suspicious cases of PTA and PTC, and serum calcium assay should be performed as a routine screening procedure. Serum calcium and PTH assays are both reliable methods for the diagnosis of PTA and PTC. A combination of ultrasonography and 99mTc-MIBI scintigraphy is sufficient for locating adenomas. Accompanied by intraoperative pathological examination, unilateral neck exploration is an acceptable approach for patients with definitely preoperative confirmed adenoma localization.

[A study on correlation of hepatic metastasis of colorectal cancer to vascular endothelial growth factor, angiopoietin-2, and fibronectin].

OBJECTIVE: To study the correlation of hepatic metastasis of colorectal cancer to vascular endothelial growth factor (VEGF), angiopoietin- 2 and fibronectin (FN). METHODS: The transcription and expression of VEGF, angiopoietin- 2 and FN were detected by semiquantitative reverse transcription- polymerase chain reaction (SQRT- PCR) and immunohistochemical staining in the specimens from sixty patients with colorectal cancer. RESULTS: The mRNA transcription and expression levels of VEGF, angiopoietin- 2 and FN in colorectal cancer tissues were obviously higher than those in paratumor normal tissues P< 0.05. The transcription and expression levels of VEGF were correlated with tumor invasion P< 0.05,Dukes' stage P< 0.05,and lymph node and/or hepatic metastasis P< 0.05. Angiopoietin expression and transcription levels were correlated with tumor differentiation. The expression of FN in extra cellular matrix (ECM) was significantly higher (P< 0.05),whereas ECM in basement membrane was significantly lower in cancer tissues than that in paratumor normal tissues (P< 0.05),which both were correlated with tumor invasion P< 0.05,Dukes' stage P< 0.05,and lymph node metastasis P< 0.05. Absence of FN protein in basement membrane was also correlated with hepatic metastasis. CONCLUSION: Colorectal cancer cells can secrete VEGF and contribute to metastasis and proliferation of tumor by stimulating the growth of tumor vessel. Both of VEGF and angiopoietin- 2 contribute to angiogenesis and the decrease of FN in basement membrane of cancer tissue is an important primary factor of hepatic metastasis.

[Correlation between the activation of AP-1 signal transduction pathway and metastasis of colorectal carcinoma].

OBJECTIVE: To investigate the binding activity of activator protein-1 (AP-1) with DNA probe in the colorectal carcinoma (CRC) tissues and surrounding tissues and explore the correlation between the activation of AP-1 signal transduction pathway and metastasis of CRC. METHODS: The AP-1 DNA binding activities were investigated by electrophoretic mobility shift assay (EMSA) in CRC specimens (T), surrounding tissues including 2 cm (P(2)), 5 cm(P(5)) far away from primary tumor margin and distal resection margin of the specimens (N). The mRNA expression level of vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9) were measured by quantitive reverse transcription polymerase chain reaction (Q- RT-PCR). RESULTS: The AP-1 DNA binding activity in T was significantly higher than those in P(2), P(5) and N (P< 0.05) tissues. There were significantly positive correlations between AP-1 DNA binding activity in tumor and invasive degree, lymphatic metastasis respectively (P< 0.01), but no correlation with histological classification and differentiation (P> 0.05). The transcription levels of VEGF and MMP-9 in CRC were significantly higher than those in P(5) and N (P< 0.01, P< 0.05) tissues. The transcription levels of VEGF and MMP-9 were significantly correlated with increasing AP-1 DNA binding activity (P< 0.01). CONCLUSIONS: AP-1 is significantly correlated to the invasion and metastasis in CRC. The activation of AP-1 signal transduction pathway might be involved in the angiogenesis and of degradation extracellular matrix during tumor metastasis.