[Effects and clinical significance of NLRP3 inflammasome activated by IL-17A in CRSwNP].

Objective: To investigate the effects and clinical significance of NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome activated by interleukin (IL)-17A in chronic rhinosinusitis with nasal polyps (CRSwNP). Methods: Patients underwent nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-sen University from January 2020 to December 2021 were collected, including 28 CRSwNP (including 19 males and 9 females, aged 19 to 67 years), 22 chronic rhinosinusitis without nasal polyps (CRSsNP) and 22 controls. qRT-PCR was used to detect the expressions of IL-17A, NLRP3, IL-1ß and IL-18 in the three groups, and their correlations were analyzed. The positions of IL-17A, NLRP3 and IL-18 in nasal polys were analyzed by immunofluorescence. Western Blotting and ELISA were employed to detect the expression of NLRP3, IL-1ß and IL-18 in the human nasal epithelial cells after using IL-17A stimulation or IL-17A receptor inhibitor. Immunofluorescence was used to observe the NLRP3, IL-1ß, and IL-18 protein expression after IL-17A stimulating human nasal epithelial cells, and after the use of IL-17A receptor inhibitor and NLRP3 inhibitor MCC950. The correlations between NLRP3, IL-1ß, IL-18 and CT scores, nasal endoscopic scores, visual analogue scale (VAS) scores, and sino-nasal outcome test (SNOT) 22 scores of CRSwNP patients were analyzed. SPSS 20.0 software was used for statistical analysis. Results: The expressions of IL-17A, NLRP3, IL-1ß and IL-18 in the tissues of CRSwNP patients were significantly higher than those in CRSsNP group(P=0.018,P<0.001,P=0.005, P=0.016) and the control group(all P<0.001). IL-17A was positively correlated with the expression of NLRP3, IL-1ß, and IL-18(r ralue was 0.643,0.650,0.629,respectively, all P<0.05). IL-17A, NLRP3, and IL-18 were co-localized in the epithelial propria of polyp tissue. IL-17A stimulated the expressions of NLRP3, IL-1ß, and IL-18 in human nasal epithelial cells. After the use of IL-17A receptor inhibitor, the expressions of NLRP3, IL-1ß, and IL-18 were significantly down-regulated. After the use of NLRP3 inhibitor MCC950, IL-17A was significantly down-regulated to promote the expression of NLRP3, IL-1ß, and IL-18. The expressions of NLRP3, IL-1ß and IL-18 were positively correlated with CT, nasal endoscopy, VAS, and SNOT22 scores in patients with CRSwNP. Conclusions: IL-17A promotes the release of IL-1ß and IL-18 by activating the NLRP3 inflammasome and aggravates the severity of the disease in CRSwNP.

[The expression profile and potential regulatory mechanism of ACE2 in chronic rhinosinusitis with nasal polyps].

Objective: To preliminarily analyze the expression of angiotensin-converting enzyme 2 (ACE2) and to investigate its potential regulatory mechanism in chronic rhinosinusitis with nasal polyps (CRSwNP). Methods: Patients underwent nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-sen University from February 2020 to May 2021 were selected, including 17 males and 6 females, aging from 23 to 66 years old. Expression of ACE2 was evaluated via immunohistochemical staining in controls with non-chronic rhinosinusitis, non-eosinophilic CRSwNP (non-ECRSwNP), and eosinophilic CRSwNP (ECRSwNP) tissue, respectively. Correlations between ACE2 and the indicated Th1/Th2-related cytokines (IFN-γ, IL-4, IL-5, IL-13, IL-25, IL-33, TSLP and periostin) were analyzed based on GSE72713 dataset. Protein-protein interaction (PPI) network was constructed via string database, immune infiltration of GSE72713 dataset was evaluated using cibersort algorithm. ACE2 was comprehensively analyzed by microRNA regulatory network, gene set enrichment analysis (GSEA) and pharmacological analysis. Statistical analysis was performed using GraphPad 7.0 and SPSS 20.0 software. Results: ACE2 was up-regulated in non-ECRSwNP compared with ECRSwNP. Microarray analysis showed that ACE2 was positively correlated with IFN-γ while inversely correlated with IL-5, IL-13 and periostin significantly. Analysis of immune infiltration suggested that ACE2 expression correlated positively with the number of M1 macrophage while negatively with M2 macrophage. GSEA demonstrated that interferon-related signaling pathways were up-regulated in non-ECRSwNP, and miRNA-200B/miRNA-200C/miRNA-429 pathways targeting ACE2 were enriched in ECRSwNP. Results of pharmacological analysis indicated that ampicillin was able to promote the expression of ACE2 whereas acetaminophen could down regulated the expression of ACE2. Conclusion: Expression pattern of ACE2 is varied in non-ECRSwNP and ECRSwNP, which may be related to the different infiltration of indicated cytokines and different regulatory pathways of miRNA.

[Evaluation of contrast-enhanced ultrasound in the differential diagnosis of benign and malignant lateral cervical lymph nodes].

Objective: To investigate the value of contrast-enhanced ultrasound (CEUS) in the diagnosis of benign and malignant lateral cervical lymph nodes. Methods: One hundred and fifteen cases of suspected lymph nodes disease, 60 males, 55 females, between November 2018 and June 2019 from clinic and inpatient in Fujian Cancer Hospital were collected. Routine ultrasound and CEUS were performed in 115 cases of lateral cervical lymph nodes. All cases underwent ultrasound-guided biopsy, and were divided into two groups of benign and malignant lymph nodes according to pathological results. The differences between the two groups were analyzed and statistically analyzed. The rate and frequency were used to qualitatively describe the ultrasound performance and observed lymph node morphology between the two groups of routine ultrasound and CEUS. The sensitivity, specificity, accuracy, negative predictive value and positive predictive value were used to evaluate the efficacy of routine ultrasound and CEUS. Results: There were 37 cases of benign lymph nodes and 78 cases of malignant lymph nodes. In benign lymph nodes, 51.4% (19/37) showed uniform high enhancement, and 48.6% (18/37) showed uneven or circular enhancement with clear boundaries and no focal enhancement area. In malignant lymph nodes, 94.9% (74/78) showed uneven enhancement, and only 5.1% (4/78) showed uniform enhancement. The difference of ultrasonic contrast medium uniformities in differentiating benign from malignant was statistically significant (P<0.001). The sensitivity, accuracy, negative predictive value and positive predictive value of routine ultrasound to differentiate benign and malignant lymph nodes were 64.1%, 65.2%, 47.2%, and 66.7%, respectively, while the corresponding values of CEUS were 93.6%, 75.7%, 73.7% and 83.9%. Conclusion: CEUS may provide a valuable basis for the differential diagnosis of benign and malignant lateral cervical lymph nodes.

[Clinical application of self-made drainage tubes in different layers of soft tissue for negative-pressure wound therapy in 33 patients].

From January 2014 to June 2018, 28 patients with different types of deep soft tissue injury or infection were admitted to the Affiliated Jiangyin Hospital of Medical College of Southeast University; 5 patients were admitted to the Zhengzhou First People's Hospital. There were 24 males and 9 females, aged 18-89 (40±20) years. Disposable suction tubes with holes cut on side walls were used as self-made drainage tubes. The authors placed the self-made drainage tubes on different deep soft tissue layers and wound surfaces after debridement. The effective drainage sections of the wound surface drainage tubes were wrapped with silver ion antimicrobial functional active dressings. Bio-permeable membrane was used to close the operative area. The drainage tubes in the deep layer of wound and wound surface were connected in parallel by a tee and connected to wall-hanging medical negative-pressure suction device to conduct negative-pressure wound treatment at -20.0 to -10.6 kPa. The deep drainage tubes were usually removed or changed 4 or 5 days after surgery.The drainage tubes in the wound surface were synchronously replaced when removing or replacing he drainage tubes in the deep layer of wound. On 4 to 15 days after surgery, the deep drainage tubes were removed. On 8 to 25 days after surgery, the wound surface drainage tubes were removed. Then the treatment was changed to a conventional dressing change until the wounds were completely healed or the wound bed was ready for skin grafts or tissue flaps. The indwelling time of deep drainage tubes in this group of patients was (6.2±2.8) days, and the indwelling time of wound surface drainage tubes was (12.0±3.0) days. The wound healing time was (22±5) days, the hospital stay time was (29±7) days, and wound bacteria were reduced from 6 species and 11 strains before treatment to 3 species and 4 strains after treatment. No adverse events such as wound bleeding, irritative pain, and chronic sinus occurred during treatment. Twenty-three patients were followed up for 13 to 28 months, no treatment-related complications were observed.

[Application and influencing factors of endorectal ultrasound in T-stage restaging of rectal cancer following neoadjuvant therapy].

Objective: To evaluate the accuracy and influencing factors of T-stage restaging of rectal cancer following neoadjuvant therapy with endorectal ultrasonography (ERUS). Methods: In a retrospective study, endorectal ultrasound was performed in 86 patients with rectal cancer following neoadjuvant therapy. The imaging results were compared with postoperative pathological T-stage. Results: The accuracy of overall T-stage restaging with ERUS was 67.4% (58/86). Additionally, the accuracy of restaging in middle and high rectal cancer was higher, with an accuracy of 76.1%(35/46)and 100%(4/4) respectively. Univariate analysis showed that the location of tumors was an independent factor affecting the accuracy of ERUS(P=0.033). Conclusion: ERUS is an effective method to restage T-stage of rectal cancer following neoadjuvant therapy.

[Effect of peripheral bloodgenomic DNA methylation on the relationship between methyl donor status and risk of breast cancer].

Objective: To explore the effect of peripheral blood genomic DNA methylation on the relationship between methyl donor status and risk of breast cancer. Methods: A case-control study was conducted. Each three hundred breast cancer cases and controls were consecutively recruited. Food frequency questionnaire was used to collect dietary information. Amounts on folate, methionine, choline and betaine intake were calculated. Blood samples were collected for DNA extraction. Peripheral blood genomic DNA methylation was measured by using the Methyl Flash(TM) Methylated DNA Quantification Kit. Pathway analysis was used to examine the effect of genomic DNA methylation on the relations between methyl donor status and risk of breast cancer. Results: The genome DNA methylation rates were 0.46%±0.25% and 0.53%±0.34%, respectively on both cases and controls, with differences statistically significant (P<0.01). Results from the pathway analysis, results showed that methionine consumption was related to genomic DNA methylation (ß=0.065, P<0.05) while genomic DNA methylation was related to the risk of breast cancerk (ß=-0.027, P<0.05), respectively. Conclusions: The level of peripheral blood genomic DNA methylation in breast cancer cases was significantly lower than that in the controls. Genomic DNA methylation seemed to have played a mediated role between methionine and the risk of breast cancer.

[Application of three-dimensional printing personalized acetabular wing-plate in treatment of complex acetabular fractures via lateral-rectus approach].

Objective: To investigate reduction and fixation of complex acetabular fractures using three-dimensional (3D) printing technique and personalized acetabular wing-plate via lateral-rectus approach. Methods: From March to July 2016, 8 patients with complex acetabular fractures were surgically managed through 3D printing personalized acetabular wing-plate via lateral-rectus approach at Department of Orthopedics, the Third Affiliated Hospital of Southern Medical University. There were 4 male patients and 4 female patients, with an average age of 57 years (ranging from 31 to 76 years). According to Letournel-Judet classification, there were 2 anterior+ posterior hemitransverse fractures and 6 both-column fractures, without posterior wall fracture or contralateral pelvic fracture. The CT data files of acetabular fracture were imported into the computer and 3D printing technique was used to print the fractures models after reduction by digital orthopedic technique. The acetabular wing-plate was designed and printed with titanium. All fractures were treated via the lateral-rectus approach in a horizontal position after general anesthesia. The anterior column and the quadrilateral surface fractures were fixed by 3D printing personalized acetabular wing-plate, and the posterior column fractures were reduction and fixed by antegrade lag screws under direct vision. Results: All the 8 cases underwent the operation successfully. Postoperative X-ray and CT examination showed excellent or good reduction of anterior and posterior column, without any operation complications. Only 1 case with 75 years old was found screw loosening in the pubic bone with osteoporosis after 1 month's follow-up, who didn't accept any treatment because the patient didn't feel discomfort. According to the Matta radiological evaluation, the reduction of the acetabular fracture was rated as excellent in 3 cases, good in 4 cases and fair in 1 case. All patients were followed up for 3 to 6 months and all patients had achieved bone union. According to the modified Merle D'Aubigné and Postel scoring system, 5 cases were excellent, 2 cases were good, 1 case was fair. Conclusions: Surgical management of complex acetabular fracture via lateral-rectus approach combine with 3D printing personalized acetabular wing-plate can effectively improve reduction quality and fixation effect. It will be truly accurate, personalized and minimally invasive.

[Value of dual contrast-enhanced ultrasound in the preoperative T staging of rectal carcinoma].

Objective: To explore the value of dual contrast-enhanced ultrasound in preoperative T staging of rectal carcinoma. Methods: Dual contrast-enhanced ultrasound examinations were performed on 72 patients with rectal carcinoma via transrectal infusion and intravenous injection. The accordance of preoperative dual contrast-enhanced ultrasound results and postoperative pathologic results was evaluated retrospectively. Results: The overall accordance rate of preoperative T staging was 73.6% (53/72). And accordance rate was 100.0% (3/3), 100.0% (5/5), 68.4% (13/19), 71.4% (25/35)and 70.0% (7/10) for Tis , T1, T2, T3 and T4, respectively. The consistency was good (κ=0.607, χ(2) =8.363, P<0.01). The accordance rate of middle/lower vs high rectal carcinoma was 68.7% and 85.7%. Conclusion: Dual contrast-enhanced ultrasound can provide reference for preoperative T staging for patients with rectal carcinoma.

Influence of Hydroxyethyl Starch on Renal Function After Orthotopic Liver Transplantation.

BACKGROUND: Postoperative renal impairment (RI) is one of the most common complications in orthotopic liver transplantation (OLT), and it occurs in 17% to 95% of the patients who undergo the surgery. METHODS: We reviewed 394 consecutive patients who underwent OLT. On the basis of the preoperative renal function level (presence of renal failure (RF): SCr >1.5 mg/dL before OLT), the patients were divided into an RF group and a non-RF group. In each group, the patients were subdivided into 4 subgroups according to the type and dosage of the intra-operative use of HES (hydroxyethyl starch). The changing tendency of the SCr (serum creatinine) of each group and the ratio of the change in the SCr within the first postoperative week were compared. RESULTS: In total, 139 of 394 patients (35%) had RI within the first week after OLT (RI group); 104 patients (75%) in the RI group and 181 patients (71%) in the non-RI group required HES transfusions. The multivariate logistic regression analysis identified old age, a low pre-operative platelet level, and massive red blood cell transfusions as risk factors for the postoperative development of RI. The changing tendency of the SCr and the ratio of change in the SCr among the different HES subgroups showed no significant difference in the RF group or in the non-RF group. CONCLUSIONS: Perioperative use of HES 200/0.5 or HES 130/0.4 has no significant effect on renal function in the first postoperative week in patients undergoing OLT.

Spinal protein kinase Mζ contributes to the maintenance of peripheral inflammation-primed persistent nociceptive sensitization after plantar incision.

BACKGROUND: Previous studies suggest that persistent post-surgical pain (PPSP) is correlated with preoperative pain status and amplification of central sensitization. Protein kinase Mζ (PKMζ) is an essential substrate of the late long-term potentiation underlying central sensitization, which is one mechanism of pain memory formation. However, the potential contributions of spinal PKMζ to PPSP, a condition in which preoperative pain is prevalent, are not known. METHODS: Here, a modified 'hyperalgesia priming' model was established to simulate the clinical situation. This model used intraplantar injections of carrageenan (Car) as priming stimuli to elicit persistent nociceptive sensitization after plantar incision in rats. Upon treatment with PKMζ inhibitor ZIP, Scr-ZIP or protein kinase Cs (PKCs) inhibitor NPC-15437, altered behaviour and spinal PKMζ/PKCs expression were observed. RESULTS: A long-lasting hypersensitivity induced by Car-priming was identified and precipitated by subsequent plantar incision in this 'two-hit' paradigm. Post-treatment with ZIP, but not Scr-ZIP and NPC-15437, after the resolution of Car-priming selectively relieved hypersensitivity. In contrast, pre-priming NPC-15437 treatment only prevented Car-induced initial transient hyperalgesia. Immunoassays showed a significant decrease in spinal PKMζ expression after plantar incision with post-priming ZIP treatment as compared with Scr-ZIP and NPC-15437, but no notable differences in PKCs expression were observed. CONCLUSIONS: Spinal PKCs solely contribute to the initial induction of persistent pain, whereas PKMζ plays an essential role in spinal plasticity storage. PKMζ is responsible for the maintenance of peripheral inflammation-primed PPSP. Therefore, spinal PKMζ may be a therapeutic target to prevent surgery-induced chronic pain in patients with preoperative pain.

Patients with IgA nephropathy have increased serum galactose-deficient IgA1 levels.

Immunoglobulin A (IgA) nephropathy is the most prevalent form of glomerulonephritis worldwide. A renal biopsy is required for an accurate diagnosis, as no convenient biomarker is currently available. We developed a serological test based upon the observation that this nephropathy is characterized by undergalactosylated IgA1 in the circulation and in mesangial immune deposits. In the absence of galactose, the terminal saccharide of O-linked chains in the hinge region of IgA1 is terminal or sialylated N-acetylgalactosamine. A lectin from Helix aspersa, recognizing N-acetylgalactosamine, was used to develop an enzyme-linked immunosorbent assay that measures galactose-deficient IgA1 in serum. The median serum lectin-binding IgA1 level was significantly higher for 153 Caucasian adult patients with IgA nephropathy without progression to end-stage renal disease as compared with that for 150 healthy Caucasian adult controls. As the lectin-binding IgA1 levels for the controls were not normally distributed, the 90th percentile was used for determination of significant elevation. Using a value of 1076 U/ml as the upper limit of normal, 117 of the 153 patients with IgA nephropathy had an elevated serum lectin-binding IgA1 level. The sensitivity as a diagnostic test was 76.5%, with specificity 94%; the positive predictive value was 88.6% and the negative predictive value was 78.9%. We conclude that this lectin-binding assay may have potential as a noninvasive diagnostic test for IgA nephropathy.

A sterically stabilized immunolipoplex for systemic administration of a therapeutic gene.

A sterically stabilized immunolipoplex (TsPLP), containing an antitransferrin receptor single-chain antibody fragment (TfRscFv)-PEG molecule, has been developed to specifically and efficiently deliver a therapeutic gene to tumor cells. A postcoating preparation strategy was employed in which a DNA/lipid complex (lipoplex) was formed first and then sequentially conjugated with PEG and TfRscFv. The complex prepared by this method was shown to be superior in ability to deliver genes to tumor cells than when prepared by a common precoating strategy, in which DNA is mixed with TfRscFv-PEG conjugated liposome. Using prostate cancer cell line DU145, a comparison was made between the in vitro and in vivo gene delivery efficiencies of four complexes, Lipoplex (LP), PEG-Lipoplex (PLP), TfRscFv-PEG-Lipoplex (TsPLP) and our standard TfRscFv-Lipoplex (TsLP). In vitro, the order of transfection efficiency was TsLP>LP approximately TsPLP>PLP. However, in vivo the order of transfection efficiency, after systemic administration via the tail vein, was TsPLP>TsLP>LP or PLP with TsPLP-mediated exogenous gene expression in tumor being two-fold higher than when mediated by TsLP. This suggests that the in vitro transfection efficiency of TsPLP was not indicative of its in vivo efficiency. In addition, it was found that the level of exogenous gene expression in the tumor mediated by TsPLP was higher than that mediated by TsLP and did not decrease over the time. More importantly, high exogenous gene expression in tumor, but low expression in liver, was observed after an i.v. delivery of TsPLP carrying either the GFP reporter gene or the p53 gene, indicating that tumor preferential targeting was maintained by this complex in the presence of PEG. These findings show that incorporation of PEG into our targeted lipoplex results in a more efficient delivery of the complex to the tumor cells, possibly by inhibiting the first pass clearance observed with non-PEG containing liposomes. Therefore, these data demonstrate that TsPLP is a improvement over our previously established tumor targeted gene delivery complex for systemic gene therapy of cancer.

Anesthetic management for separation of craniopagus twins.

We report the anesthetic management of a case of separation of craniopagus twins with unbalanced cross circulation and one twin with renal dysfunction. After intravenous induction, anesthesia was maintained with isoflurane inhalation and propofol infusion. Twin A survived but Twin B died after the surgery. The anesthetic problems during the operation are discussed.

The influence of anaesthesia and surgery on the circadian rhythm of melatonin.

BACKGROUND: Operations are typically associated with sleep and other circadian rhythm disturbances. The present study was set up to evaluate the influence of spinal and general anaesthesia associated with knee surgery on the circadian rhythm of melatonin, which has sleep inducing properties. Previously this context has been studied only in some invasive operations and it might be that general anaesthesia induces more disturbances on circadian rhythm of melatonin than operations done with patients awake. METHODS: The circadian secretion pattern of melatonin was monitored during the pre- and postoperative evenings, nights and mornings to clarify possible anaesthesia/surgery-induced changes in the nocturnal secretion of melatonin and in the phase of the melatonin rhythm. The study included 20 patients scheduled for minor orthopaedic operations. The patients were randomised to receive either spinal or general anaesthesia. Melatonin was measured from evening and morning saliva samples radioimmunologically. The nocturnal urine before and after surgery was radioimmunologically examined for 6-hydroxymelatonin sulphate. RESULTS: Melatonin secretion evaluated from the saliva samples was significantly diminished during the first postoperative evening as compared with that during the preoperative evening (P<0.001). There was also a significant decline of 26% (P<0.05) in postoperative 6-hydroxymelatonin sulphate excretion. There was no significant difference in melatonin secretion between the spinal and general anaesthesia groups. CONCLUSION: Our findings suggest that anaesthesia in conjunction with surgery acutely disturbed the normal circadian rhythm of melatonin by delaying the onset of nocturnal melatonin secretion.

Bone marrow endothelial cells secrete thymosin beta4 and AcSDKP.

Bone marrow endothelial cells are the essential component of the bone marrow microenvironment. They produce many kinds of cytokines, including stimulators and inhibitors. Many researchers have suggested that in the presence of endothelial cell layer, CD34+CD38- cells are capable of expansion. The ability of the endothelial cell layer to protect hematopoietic stem cells from extensive differentiation may be related to the inhibitors derived from endothelial cells. The aim of the present study was to determine whether the inhibitors thymosin beta4 and AcSDKP are elaborated by murine bone marrow endothelial cells. Murine bone marrow endothelial cells (mBMECs) were cultured in serum-free conditioned medium. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to analyze the differential expression of the thymosin-beta gene, and reverse phase high-performance chromatography (HPLC) and mass spectroscopy were used to determine the concentration of thymosin beta4 (Tbeta4) and AcSDKP in EC lysate and in the medium (mBMEC-CM). Colony-forming unit granulocyte-macrophage (CFU-GM) colony assays were used to examine the effect of components (mw 3-10 kD, <3 kD) of mBMEC-CM, thymosin beta4, and AcSDKP on the proliferation of hematopoietic cells.mBMECs expressed Tbeta4 mRNA. In EC lysate and mBMEC-CM, Tbeta4 and AcSDKP were detected. After adding protease inhibitors, the concentration of Tbeta4 in EC lysate increased significantly, while the concentration of AcSDKP decreased. mBMEC-CM (mw 3-10 kD) had no effect on the formation of CFU-GM. However, mBMEC-CM (mw <3 kD) could inhibit the growth of CFU-GM. Tbeta4 (10(-11) approximately 10(-7)mol/L) and AcSDKP (10(-11) approximately 10(-5)mol/L) had dose-dependent inhibitory effects on the growth of CFU-GM. Angiotensin converting enzyme (ACE), the enzyme degrading AcSDKP, could partially eliminate the inhibitory effect of mBMEC-CM (mw <3 kD) on CFU-GM.BMECs express and secrete Tbeta4 and AcSDKP. Tbeta4 exists in the 3-10 kD component of mBMEC-CM, while AcSDKP exists in the <3 kD component of ECCM. Both components exert inhibitory effects on the proliferation of hematopoietic progenitors.

Presence of gonadotropin-releasing hormone (GnRH) and its mRNA in rat pancreas.

Gonadotropin-releasing hormone (GnRH) has been reported to exist in many non-hypothalamic tissues, such as the placenta, gonads, and mammary glands, while there still have been no reports concerning the existence and expression of GnRH in the mammalian digestive system. Immunocytochemistry and in situ hybridization results show that GnRH molecule and GnRH mRNA are both exclusively distributed in exocrine pancreas, and RT-PCR result further proves that GnRH transcription units do exist in the pancreas, which possess the same sequence as the hypothalamus GnRH mRNA. Quantitative analysis indicates that mRNA levels in rat pancreas remain at a low level (less than 10% of that in hypothalamus) without sexual or developmental difference. This is the first report suggesting the existence and gene expression of GnRH in rat pancreas.

Expression of vascular endothelial growth factor and its role in oncogenesis of human gastric carcinoma.

AIM: To establish the role of vascular endothelial growth factor (VEGF) in the oncogenesis of human gastric carcinoma more directly. METHODS: The expression of VEGF and its receptor kinase-domain insert containing receptor (KDR) in human gastric cancer tissue were observed by immunohistochemical staining. VEGF levels were manipulated in human gastric cancer cell using eukaryotic expression constructs designed to express the complete VEGF(165) complimentary DNA in either the sense or antisense orientation. The biological changes of the cells were observed in which VEGF was up-regulated or down-regulated. RESULTS: VEGF-positive rate was 50%, and VEGF was mainly localized in the cytoplasm and membrane of the tumor cells, while KDR was mainly located in the membrane of vascular endothelial cells in gastric cancer tissues and peri-cancerous tissue. In 2 cases of 50 specimens, the gastric cancer cells expressed KDR, localized in both the cytoplasm and membrane. Introduction of VEGF(165) antisense into human gastric cancer cells (SGC-7901, immunofluorescence intensity, 31.6%)) resulted in a significant reduction in VEGF-specific messenger RNA and total and cell surface VEGF protein (immunofluorescence intensity, 8.9%) (P<0.05). Conversely, stable integration of VEGF(165) in the sense orientation resulted in an increase in cellular and cell surface VEGF (immunofluorescence intensity, 75.4%) (P<0.05). Lowered VEGF levels were associated with a marked decrease in the growth of nude mouse xenografted tumor (at 33 days postimplantation, tumor volume: 345.40 +/- 136.31 mm3)(P<0.05 vs control SGC-7901 group: 1534.40 +/- 362.88 mm3), whereas up-regulation of VEGF resulted in increased xenografted tumor size (at 33 days postimplantation, tumor volume: 2350.50 +/- 637.70 mm3) (P<0.05 vs control SGC-7901 group). CONCLUSION: This study provides direct evidence that VEGF plays an important role in the oncogenesis of human gastric cancer.

Expression and function of angiopoietin-1 in breast cancer.

Angiopoietin-1 (Ang1) has been shown to act as an angiogenic promoter in embryonic angiogenesis by promoting vascular branching, pericyte recruitment and endothelial survival. We have investigated the role of Ang1 in tumour neovascularization under clinical conditions and in animal models. The expression of Ang1 in clinical breast cancer specimens was analysed by using laser-capture microdissection and reverse transcriptase-linked polymerase chain reaction (RT-PCR) on RNA isolated from the samples. Despite the expression of Ang1 in many human breast cancer cell lines, the gene was expressed in only three of 21 breast cancer clinical specimens, even though its receptor, Tie2, is abundant in the vasculature of all of these tumours. Ang1 was then overexpressed in a human breast cancer cell line (MCF-7) on its own and in conjunction with FGF1, an angiogenic factor shown to be able to increase the tumorigenicity of MCF-7 cells. High concentrations of Ang1 were produced in the conditioned media of the transfected cells (range 156-820 ng ml(-1)). However, in contrast to its physiological role as promoter of angiogenesis, overexpression of Ang1 did not enhance tumour growth, but instead caused up to a 3-fold retardation of tumour growth (P = 0.003).