[The clinical application of two-complex and one-plane technique for renal artery location in transperitoneal laparoscopic radical nephrectomy].

Objective: To examine the clinical application effects of two-complex and one-plane technique for the renal artery dissected location in transperitoneal laparoscopic radical nephrectomy(LRN). Methods: Clinical data of patients diagnosed as renal tumor between January 2016 and December 2019 that treated with transperitoneal LRN and the two-complex and one-plane technique was performed to locate and dissect the renal arteries were analyzed retrospectively. A total of 206 qualified patients were enrolled, included 71 cases from Kaifeng Central Hospital and 135 cases from the First Affiliated Hospital of Zhengzhou University. There were 126 males and 80 females with median age of 54 years (range: 35 to 82 years). Renal tumor lived at left kidney in 102 cases and at the right in 104 cases, with a maximum tumor diameter of (6.8±2.5)cm (range: 3.0 to 12.7 cm). During the procession of transperitoneal LRN, The genital veins was followed to locate renal veins, soon the renal vein complex and renal lower pole complex and psoas major muscle plane that named as "two-complex and one-plane"anatomical markers were applied to the dissected location of renal arteries. The established application of two-complex and one-plane technique for renal artery location, and the anatomical features along with its locational time of renal artery, as well as vascular-related complications were recorded and analysed. The χ2 test or t test was used to compare the results of different lateral operations. Results: The surgical procedures were successful in all 206 patients. The operation time was (54.4±13.6) minutes (range:22 to 116 minutes), no injury to liver, spleen or intestine. Two-complex and one-plane technique was used to complete renal arteries location of 206 patients that accompanied with one or more arterial branch in the procession of transperitoneal LRN. Single renal artery branch was found in 163 cases, two or more renal artery branches were found in 43 cases, and 60 branches of accessory renal artery that in addition to the main trunks were detected in 14 cases (13.7%, 14/102) on left-side and 29 cases (27.9%, 29/104) on the right(χ²=6.251, P=0.012).The main branch of renal artery that directly been located inferior or posterior to renal vein in 165 cases, and the other 41 cases that born with higher-position of main branch been located through an upper"window-opening"of renal veins complex. Of the 60 accessory renal artery, 46 branch (76.6%, 46/60) been located inferior or posterior to renal veins, and the other 14 branch with higher-position (23.3%, 14/60) been located by a"window-opening"technique. The renal artery dissected location time was (21.2±9.4) minutes (range:11 to 43 minutes) in left-side and (17.5±9.3) minutes (range:9 to 32 minutes) in the right (t=2.840, P=0.005).The intraoprative bleeding was (51.8±25.2) ml (range:20 to 400 ml). There were 4 cases of vascular injury occured and treated with laporoscopy, only one need blood transfusion. Conclusions: The anatomical markers of renal vein complex and renal lower pole complex and psoas major muscle plane that can apply to locate various anatomical position types of renal artery in transperitoneal LRN effectively. The exactly application of two-complex and one-plane technique that could shorten the operational time of renal artery location and reduce the complications of vascular injury.

[Clinical and imaging characteristics of patients with different types of infectious sacroiliitis].

To investigate the clinical manifestations and imaging characteristics of patients with different types of infectious sacroiliitis. Clinical data of 40 patients diagnosed with infectious sacroiliitis were retrospectively analyzed. Among the 40 patients, 16 patients were diagnosed as non-brucellar and non-tuberculous infectious sacroiliitis (ISI), 13 with tuberculous infectious sacroiliitis (TSI), and 11 with brucellar sacroiliitis (BSI). In the ISI and TSI group, female patients accounted for 11/16, 12/13, while the proportion of unilateral involvement was 15/16 and 12/13, respectively. Compared with ISI and TSI group, BSI patients were mainly male (8/11) and presented more bilateral involvement (6/11) (P<0.05). Bone erosion was more common in ISI and TSI groups than in BSI group (6/15, 7/11 and 2/10), as well as abscess formation (3/15, 4/11 and 1/10, respectively). Symptoms in all patients relieved 1-2 weeks after administration of antibiotics or anti-tuberculosis treatment, but the resolution of the magnetic resonance imaging findings delayed about 6 (3-9) months. ISI and TSI patients with infectious sacroiliitis should be differentiated from spondyloarthritis, with a characteristic of more female patients, unilateral sacroiliitis, bone erosion, soft tissue involvement and abscess formation. However, BSI patients are mainly male, more bilateral involvement and less bone destruction and abscess formation. Antibiotic therapy demonstrates significant therapeutic effects, but resolution of the magnetic resonance imaging findings responses late.

[Clinical characteristics and risk factors of patients with systemic lupus erythematosus and cancer].

To investigate the clinical manifestations and risk factors in patients with systemic lupus erythematosus (SLE) and cancers. From October 2010 to February 2019, 5 566 SLE patients hospitalized in the First Affiliated Hospital of Zhengzhou University were enrolled. A total of 69 cancer patients were identified, and the clinical characteristics and previous treatment were analyzed. Cervical carcinoma (21.74%, 15/69) and thyroid cancer (21.74%, 15/69) were the most common types of cancer. Most cancers were diagnosed in SLE patients with an age 40~50 years. The disease duration of SLE was from 60~120 months. SLE patients without cancers were usually diagnosed between 20~30 years with duration of symptoms less than 12 months. As to the previous treatment of SLE, the uses of glucocorticoid, cyclophosphamide, methotrexate and azathioprine were comparable between patients with cancers and without (P>0.05). However, the use of hydroxychloroquine was more frequent in SLE patients than in patients with cancers (P<0.01). Correlation analysis revealed significant correlation between disease course of SLE (OR=4.25, 95%CI 1.79~10.01,P<0.001), hydroxychloroquine (OR=0.26, 95%CI 0.12~0.59,P<0.001) and cancer risk. Long disease course may be a risk factor for SLE patients to develop cancer, whereas hydroxychloroquine could be a protective factor.

Application of High-Resolution C-Arm CT Combined with Streak Metal Artifact Removal Technology for the Stent-Assisted Embolization of Intracranial Aneurysms.

BACKGROUND AND PURPOSE: Metal artifacts from coils and stents limit the level of detail in C-arm CT images of stent attachment and coiling attenuation in the aneurysm neck. We evaluated the utility of high-resolution C-arm CT combined with streak metal artifact removal technology for stent-assisted embolization of intracranial aneurysms. MATERIALS AND METHODS: From October 2017 to July 2018, the First Affiliated Hospital of Zhengzhou University treated 107 patients with intracranial aneurysms (118 aneurysms in total) with stent-assisted embolization. Conventional C-arm CT and high-resolution C-arm CT scanning of the stented area were performed during and after treatment. 3D images were reconstructed with and without streak metal artifact removal techniques. Subsequently, the image quality was compared. The reconstructed images indicated the stent deployment degree and packing density. Follow-up assessments included clinical and angiographic outcomes and complications. RESULTS: In total, 118 aneurysms were successfully embolized using 118 stents. Image quality was significantly higher (P < .05) with high-resolution C-arm CT combined with streak metal artifact removal reconstruction. Streak metal artifact removal reconstruction and 2D angiography at working angles showed incomplete deployment of 6 stents and incomplete aneurysm embolization of 15 patients, which were subsequently resolved. One case of hemorrhage was noted postoperatively. Follow-up of 93 patients at 6-13 months indicated 3 cases of aneurysm recurrence. CONCLUSIONS: High-resolution C-arm CT combined with the streak metal artifact removal technique effectively reduced metal artifacts from stents and coils during aneurysm embolization. This method can help physicians determine the extent of stent deployment and the packing density of coils and thus potentially reduce complications and aneurysm recurrence.

CalliSpheres® drug-eluting beads (DEB) transarterial chemoembolization (TACE) is equally efficient and safe in liver cancer patients with different times of previous conventional TACE treatments: a result from CTILC study.

PURPOSE: To assess the efficacy and safety of drug-eluting beads transarterial chemoembolization (DEB-TACE) in liver cancer patients with different times of previous conventional transarterial chemoembolization (cTACE) treatments. METHODS: 367 liver cancer patients about to receive DEB-TACE treatment were enrolled in this prospective cohort study. All patients were divided into no previous cTACE group (NPC group), 1-2 times previous cTACE group (PC group) and triple or above previous cTACE group (TPC group) according to the times of previous cTACE treatments. RESULTS: There was no difference in complete response (CR) (P = 0.671) and objective response rate (ORR) (P = 0.062) among three groups. Additionally, no difference in overall survival (OS) among groups (P = 0.899) was found. As to liver function, most liver function indexes were deteriorative at 1 week after DEB-TACE operation, but returned to baseline at 1-3 months after DEB-TACE operation in all three groups, while percentage of abnormal total bile acid (TBA) patients was higher in TPC group than NPC and PC groups at 1-3 month post-DEB-TACE (P = 0.018). As for safety profiles, the incidence of pain during DEB-TACE operation was lower in TPC group compared to NPC and PC groups (P = 0.005), while no difference of other adverse events was found during and 1 month post-DEB-TACE treatment among three groups. CONCLUSION: DEB-TACE treatment was equally efficient and tolerated in liver cancer patients with different times of previous cTACE treatments.

Analysis of risk factors and prevention strategies of post-ERCP pancreatitis.

OBJECTIVE: To analyze the relevant risk factors and preventive measures for post-endoscopic retrograde cholangiopancreatography (post-ERCP) pancreatitis (PEP) so as to improve the diagnosis and treatment levels of ERCP, thus reducing the prevalence rate of PEP. PATIENTS AND METHODS: The clinical data of 278 patients receiving ERCP from January 2014 to December 2016 were retrospectively analyzed. First, the univariate analysis was conducted for the factors such as gender, age, diameter of common bile duct, whether development occurred in the pancreatic duct and other factors. Then, the multivariate logistic regression analysis was performed for factors showing statistical significance in the univariate analysis so as to analyze the independent risk factors for PEP. RESULTS: The success rate of ERCP included in the study was 97.12%. The prevalence rate of PEP was 8.27%. Univariate analysis results showed that the prevalence rate of PEP in the group of patients younger than 60 years old was higher than that in the group of patients older than 60 years old (14.14% vs. 5.03%, p = 0.016); the prevalence rate of PEP in the group of patients with intubation difficulty was higher than that in the group of patients without intubation difficulty (19.61% vs. 5.73%, p = 0.004); the prevalence rate of PEP in the group of patients with operation time more than 60 min was higher than that in the group of patients with operation time less than 60 min (18.60% vs. 6.38%, p = 0.034); the prevalence rate of PEP in the group of patients with the pancreatic duct development was higher than that in the group of patients without the pancreatic duct development (50% vs. 6.72%, p < 0.001); the prevalence rate of PEP in the group of patients placed with nasobiliary drainage catheters was not higher than that in the group of patients not placed with nasobiliary drainage catheters (18.00% vs. 2.81%, p < 0.001). The above five relevant factors were included in the logistic regression equation for the multivariate analysis, which showed that the age less than 60 years old (p = 0.002) and the pancreatic duct development (p = 0.004) were independent risk factors for PEP, and nasobiliary drainage (p = 0.003) was a protective factor for PEP. CONCLUSIONS: The occurrence of PEP is associated with the age less than 60 years old, the pancreatic duct development, intubation difficulty and overlong operation time. Among them, the pancreatic duct development and the age less than 60 years old are independent risk factors for PEP. Placing nasobiliary drainage catheters after operation, avoiding the pancreatic duct development, improving the success rate of intubation, reducing ERCP operation time and other methods, can effectively reduce the occurrence of PEP.

LncRNA SNHG7 promotes the proliferation and inhibits apoptosis of gastric cancer cells by repressing the P15 and P16 expression.

OBJECTIVE: To investigate the relative expression of long non-coding small nucleolar RNA host gene 7 (lncRNA SNHG7) in gastric cancer tissues and cells, the effect of lncRNA SNHG7 on proliferation and apoptosis of gastric cancer cells in vivo and in vitro experiments, and the relevant mechanism. PATIENTS AND METHODS: Real-time quantitative polymerase chain reaction (qRT-PCR) experiment was performed to detect the relative expressions of SNHG7 in the gastric cancer tissues and cells. In presence of lip2000, SNHG7 interference sequence was transiently transfected into the gastric cancer cells followed by transfection efficiency detection by qRT-PCR. Cell count kit 8 (CCK-8) and clone formation assay were also carried out to detect the effect of SNHG7 on the proliferation of gastric cancer cells, flow cytometry (FCM) to detect the effect of SNHG7 on the cycle and apoptotic rate of gastric cancer cells, in vivo experiment to detect the effect of SNHG7 on biological behaviors of gastric cancer cells, and Western blotting assay to detect the variations in expression of downstream proteins after SNHG7 expression was interfered. RESULTS: The qRT-PCR experiment showed that in a total of 68 cases of cancer tissues and tumor-adjacent tissues, the relative expression of SNHG7 was upregulated in 48 cases of gastric cancer tissues and 5 gastric cancer cell lines. The in vitro experiments showed that after SNHG7 expression was interfered, the proliferation of gastric cancer cells was inhibited with an increase in apoptotic rate and arrest of cell cycle in G1/G0 phase. Experiment on nude-mouse transplanted tumor model confirmed that after SNHG7 expression was interfered, in vivo tumor growth was inhibited, and the Western blotting assay revealed that regulation of p15 and p16 expressions constituted a part of the potential molecular mechanism. CONCLUSIONS: Relative expression of SNHG7 is upregulated in gastric cancer tissues and cells, and partially contributes to the development and progression of gastric cancer through regulation of p15 and p16 expressions. Thus, interference on expression of SNHG7 can provide critical the theoretical basis for inverting the malignant phenotype of gastric cancer in clinical practice.

25-hydroxyvitamin D is associated with metabolic syndrome among premenopausal women with systemic lupus erythematosus in China.

Objectives This study aimed to investigate the status of 25-hydroxyvitamin D (25(OH)D) and its association with metabolic syndrome (MS) and different MS components among premenopausal women with systemic lupus erythematosus (SLE) in China. Patients and methods Altogether 113 premenopausal women with SLE and the age-matched healthy cohorts were recruited in this cross-sectional study. Clinical manifestations and laboratory data including serum 25(OH)D concentration were collected. A multivariable analysis was performed to analyze the association of 25(OH)D with MS and its components. Results The prevalence of 25(OH)D deficiency (25(OH)D < 20 ng/ml) and MS were common (24.8% and 30.1%, respectively) in premenopausal patients with SLE in China. Analysis of the association between 25(OH)D, MS and its components demonstrated that the lower level of 25(OH)D was associated with increased MS prevalence (OR = 0.920, p = 0.012), a decreased level of high-density lipoprotein (OR = 1.059, p = 0.033) and a higher level of fasting glucose (OR = 0.810, p = 0.004). These associations were still detectible after adjustment for age, body mass index and SLE-related variables. Conclusion The level of 25(OH)D is associated with MS and its components in premenopausal women with SLE.

Severe vitamin D deficiency increases the risk for moderate to severe disease activity in Chinese patients with SLE.

OBJECTIVE: We aimed to investigate the prevalence of vitamin D deficiency in Chinese lupus patients and to assess the association between vitamin D levels and disease severity. METHODS: Serum levels of 25OHD3 in 121 patients with systemic lupus erythematosus (SLE) and 150 healthy controls were measured by electrochemiluminescence immunoassay. Data regarding demographics and clinical parameters were collected. Disease activity of SLE was evaluated according to the SLE Disease Activity Index (SLEDAI) score and irreversible organ damage by the Systemic Lupus International Collaborating Clinic/American College of Rheumatology, SLICC/ACR Damage Index (SDI). The multivariate logistic regression model was used to investigate the association between the degree of vitamin D deficiency and SLEDAI or SDI scores. RESULTS: The prevalence of vitamin D insufficiency (25OHD3 <30 ng/ml) and severe deficiency (25OHD3 <10 ng/ml) in SLE patients was 62.81% and 34.71%, respectively. Logistic regression analysis indicated that the cut-off point of 25OHD3 concentration was 10 ng/ml where its level was correlated with increased SLEDAI (OR 6.420, p = 0.006), but not with the SDI. In addition, hydroxychloroquine treatment lowered the SLEDAI increased by the severe 25OHD3 deficiency (OR 0.280, p = 0.008). Moreover, long disease duration (OR 1.014, p = 0.008) predicted moderate to severe organ damage. CONCLUSIONS: Vitamin D deficiency is highly prevalent in patients with SLE. Severe deficiency increases the risk for moderate to severe disease activity, but not for organ damage.

C-arm CT-guided renal arterial embolisation followed by radiofrequency ablation for treatment of patients with unresectable renal cell carcinoma.

AIM: To explore the value of using flat detector (FD) equipped angiographic C-arm CT (CACT) systems in treating unresectable renal cell carcinoma (RCC) by selective renal arterial embolisation (RAE) followed by radiofrequency ablation (RFA) (RAE-RFA). MATERIALS AND METHODS: A total of 28 patients who were not candidates for surgery were enrolled. The average size of tumours was 6.7±2.2 cm (range 4.1-9.6 cm). Twenty-eight tumours were treated with CACT-guided RFA, 5-7 days after CACT-guided RAE. RESULTS: CACT-guided RAE-RFA was technically successful in all patients. Tumour enhancement disappeared after a single RAE-RFA session in 20 patients, after two RAE-RFA sessions in four patients and after three RAE-RFA sessions in the other four patients. One patient died of lung metastasis and haematuria 13 months after RAE-RFA, and another patient died of pulmonary heart disease 23 months after repeat RAE-RFA. In the 26 living patients, tumours remained controlled during a mean follow-up period of 27 months and showed significant reduction in tumour size (6.7±2.2 cm to 3.9±1.7 cm, p<0.01). There were no significant changes in creatinine levels or urea nitrogen concentrations before and after the last RAE-RFA (p>0.05). There were no serious complications during and after the procedure. CONCLUSION: CACT-guided RAE followed by RFA appears to be a safe and effective technique for treating patients with inoperable RCC.

Clinical and cytogenomic studies in a case of infertility associated with a nonmosaic dicentric Y chromosome.

In this study, a short stature male with infertility is reported. Semen analysis and serum concentrations of FSH, LH, T and PRL were estimated. Chromosome analysis was performed on lymphocytes obtained from both the male and his parents. Cytogenomic studies were performed by fluorescent in situ hybridisation and the CytoScan(™)  HD array analysis to detect Y chromosomal rearrangements and copy number mutations. Semen analysis showed severe oligozoospermia. Numerous spermatogenic cells were observed in the semen, and approximately 60% of the cells examined in semen were primary spermatocytes, showing spermatogenic arrest at the primary spermatocyte level. Cytogenomic studies of blood revealed his karyotype which was 46,X,i(Y) (p11.32) (Yqter→Yp11.32::Yp11.32→Yqter).ish (DYZ3++, SRY++, SHOX-). array (PLCXD1→SHOX) ×1,(SRY →GOLGA2P3Y)×2, (DHRSX→ ASMT, SPRY3 →IL9R)×3. The rearrangement Y chromosome is de novo. This is the first case reported with a nonmosaic 46,X, i (Y) (p11.32), which will be useful to estimate the infertility phenotype-molecular karyotype correlation. Haploinsufficiency of short stature homeobox-containing gene is primarily responsible for the short stature. Aberrations in pseudoautosomal region 1 on the rearranged Y chromosome may result in the deficiency of X-Y pairing or recombination, ultimately lead to the spermatogenic failure.

BMP-2 induces ATF4 phosphorylation in chondrocytes through a COX-2/PGE2 dependent signaling pathway.

OBJECTIVE: Bone morphogenic protein (BMP)-2 is approved for fracture non-union and spine fusion. We aimed to further dissect its downstream signaling events in chondrocytes with the ultimate goal to develop novel therapeutics that can mimic BMP-2 effect but have less complications. METHODS: BMP-2 effect on cyclooxygenase (COX)-2 expression was examined using Real time quantitative PCR (RT-PCR) and Western blot analysis. Genetic approach was used to identify the signaling pathway mediating the BMP-2 effect. Similarly, the pathway transducing the PGE2 effect on ATF4 was investigated. Immunoprecipitation (IP) was performed to assess the complex formation after PGE2 binding. RESULTS: BMP-2 increased COX-2 expression in primary mouse costosternal chondrocytes (PMCSC). The results from the C9 Tet-off system demonstrated that endogenous BMP-2 also upregulated COX-2 expression. Genetic approaches using PMCSC from ALK2(fx/fx), ALK3(fx/fx), ALK6(-/-), and Smad1(fx/fx) mice established that BMP-2 regulated COX-2 through activation of ALK3-Smad1 signaling. PGE-2 EIA showed that BMP-2 increased PGE2 production in PMCSC. ATF4 is a transcription factor that regulates bone formation. While PGE2 did not have significant effect on ATF4 expression, it induced ATF4 phosphorylation. In addition to stimulating COX-2 expression, BMP-2 also induced phosphorylation of ATF4. Using COX-2 deficient chondrocytes, we demonstrated that the BMP-2 effect on ATF4 was COX-2-dependent. Tibial fracture samples from COX-2(-/-) mice showed reduced phospho-ATF4 immunoreactivity compared to wild type (WT) ones. PGE2 mediated ATF4 phosphorylation involved signaling primarily through the EP2 and EP4 receptors and PGE2 induced an EP4-ERK1/2-RSK2 complex formation. CONCLUSIONS: BMP-2 regulates COX-2 expression through ALK3-Smad1 signaling, and PGE2 induces ATF4 phosphorylation via EP4-ERK1/2-RSK2 axis.

Percutaneous transhepatic cholangiobiopsy to determine the pathological cause of anastomotic stenosis after cholangiojejunostomy for malignant obstructive jaundice.

AIM: To investigate the feasibility and advantages of cholangiobiopsy during percutaneous transhepatic cholangiography in the histopathological diagnosis of anastomotic stenosis after cholangiojejunostomy for malignant obstructive jaundice. MATERIALS AND METHODS: Using biopsy forceps, specimens were collected from the site of stenosis in patients with recurrent jaundice (n = 24) who had previously undergone cholangiojejunostomy for malignant obstructive jaundice. RESULTS: Stenosis occurred in all patients at the biliary-enteric anastomosis based on percutaneous transhepatic cholangiography, and was the location of the biopsy. Satisfactory specimens were obtained from 22 out of 24 patients. The sensitivity was 91.7% (22/24). Tumour tissue was obtained in 18 cases, confirming disease recurrence. Histopathological changes in four patients were diagnosed as fibroplasia and/or inflammation. These were considered cicatricial stenoses based on histopathological, imaging, and laboratory findings. The remaining two histopathology-negative patients were proven to have recurrent tumour based on imaging, laboratory, and follow-up data. No complications occurred during biopsy, including gastrointestinal haemorrhage or perforation. Either cholangial drainage and/or an inner stent was used following biopsy, which resulted in a noticeable decrease in jaundice postoperatively (p < 0.05). CONCLUSION: Percutaneous transhepatic cholangiobiopsy using biopsy forceps for the diagnosis of anastomotic stenosis after cholangiojejunostomy for malignant biliary obstructive jaundice is easy to perform and safe, with a high level of sensitivity. Interventional therapies, such as percutaneous transhepatic cholangial drainage and stent placement, can be performed concurrently, markedly improving the symptoms of patients with obstructive jaundice.

Putative signaling action of amelogenin utilizes the Wnt/beta-catenin pathway.

BACKGROUND AND OBJECTIVE: While it has long been known that amelogenin is essential for the proper development of enamel, its role has generally been seen as structural in nature. However, our new data implicate this protein in the regulation of cell signaling pathways in periodontal ligament cells and osteoblasts. In this article we report the successful purification of a recombinant mouse amelogenin protein and demonstrate that it has signaling activity in isolated mouse calvarial cells and human periodontal ligament cells. MATERIAL AND METHODS: To determine the regulatory function of canonical Wnt signaling by amelogenin, we used TOPGAL transgenic mice. These mice express a beta-galactosidase transgene under the control of a LEF/TCF and beta-catenin-inducible promoter. To investigate in greater detail the molecular mechanisms involved in the beta-catenin signaling pathway, isolated osteoblasts and periodontal ligament cells were exposed to full-length recombinant mouse amelogenin and were evaluated for phenotypic changes and beta-catenin signaling using a TOPFLASH construct and the LacZ reporter gene. RESULTS: In these in vitro models, we showed that amelogenin can activate beta-catenin signaling. CONCLUSION: Using the TOPGAL transgenic mouse we showed that amelogenin expression in vivo is localized mainly around the root, the periodontal ligament and the alveolar bone.

Involvement of a disintegrin and a metalloproteinase 8 (ADAM8) in osteoclastogenesis and pathological bone destruction.

OBJECTIVES: The eventual role of a disintegrin and a metalloproteinase 8 (ADAM8) in osteoclastogenesis was studied in erosive rheumatoid arthritis (RA) and in vitro. METHODS: ADAM8 protein and mRNA expression was measured in RA pannus and synovitis and compared to osteoarthritic (OA) synovial membrane. Human monocytes were isolated and stimulated with proinflammatory cytokines and their ADAM8 expression and surface ADAM8 were measured. Human peripheral blood monocytes and RAW 264.7 mouse monocyte/macrophage cells were stimulated to osteclast like-cells, and their expression of ADAM8 and osteoclastic markers (calcitonin receptor, integrin beta 3, cathepsin K, TRAP) were analysed. Transfection and small interfering RNA (siRNA) were used to assess the role of ADAM8 in formation of polykaryons. RESULTS: Increased numbers of ADAM8 positive cells were shown particularly in the pannus-cartilage/bone junction close or adjoining to TRAP positive multinucleate cells under formation (60 (2)% in pannus, 47 (2)% in synovitis vs 10 (1)% in OA, p<0.001). Human pannus contained high ADAM8 mRNA copy numbers (23 (7) in pannus, 14 (4) in synovitis vs 1.7 (0.3) in OA, p<0.001). Functional studies in vitro disclosed ADAM8 mRNA and protein, which was first converted to a proteolytically active and then to fusion-active form. Gene transfection and siRNA experiments enhanced and inhibited, respectively, expression of osteoclast markers and maturation of multinuclear cells. CONCLUSIONS: ADAM8 may be involved in bone destruction in RA because it is upregulated in RA pannus adjacent to developing erosions and enhances maturation of osteoclast-like cells.

IGF-1 regulation of type II collagen and MMP-13 expression in rat endplate chondrocytes via distinct signaling pathways.

OBJECTIVE: Abnormal maturation and ossification of the endplate chondrocytes play a central role in the pathogenesis of degenerative disorders of the cervical spine. It is widely held that insulin like growth factor-1 (IGF-1) stimulates chondrocyte proliferation and inhibits chondrocyte terminal differentiation both in vitro and in vivo. However, the mechanism underlying such regulation is not fully understood. The present study aimed to determine the role of IGF-1 on the mRNA expression of collagen type II, alpha 1 (Col2a1) and matrix metallopeptidase 13 (MMP-13) in rat endplate chondrocytes. The possible pathways that transduce IGF-1 effects such as phosphatidylinositol-3 (PI-3)-kinase (PI3K) and mitogen activated protein kinase (MAPK) were also investigated in these cells. METHODS: Cultured endplate chondrocytes harvested from rat cervical spines were treated with IGF-1 (100ng/ml), and the changes in Col2a1 and MMP-13 mRNA were monitored with real-time polymerase chain reaction (PCR). MMP-13 activity was also assayed. Activation of signaling proteins was evaluated by western blot analysis. Cells were also treated with pharmacological agents that block PI3K and MAPK signaling pathways. RESULTS: IGF-1 increased Col2a1 mRNA expression in rat endplate chondrocytes in a time- and dose-dependent manner. IGF-1 treatment resulted in a fourfold increase of Col2a1 mRNA with the effect maximizing at 24h. In contrast, IGF-1 treatment for 24h caused a roughly 50% reduction in MMP-13 mRNA. Similar effects were seen on the protein levels of type II collagen (col2) and MMP-13. Consistent with these results, IGF-1 also repressed MMP-13 activity. IGF-1 activated both the PI3K and the extracellular signal-regulated kinase (ERK) pathways as evidenced by phosphorylation of either Akt or ERK1/2 (respectively). The PI3K inhibitor Wartmannin significantly inhibited the IGF-1 effect on Col2a1 mRNA expression but did not affect IGF-1-induced repression of MMP-13 expression. In contrast, the ERK/MAPK inhibitor PD98059 significantly inhibited the effect of IGF-1 on MMP-13 mRNA repression and enhanced IGF-1-induced Col2a1 mRNA expression. CONCLUSIONS: In rat endplate chondrocytes the PI3K pathway mainly transduces IGF-1 effect on col2 expression while the ERK pathway mediates IGF-1 effect on MMP-13 expression.

RANKL and cathepsin K in diffuse sclerosing osteomyelitis of the mandible.

BACKGROUND: Diffuse sclerosing osteomyelitis (DSO) of the mandible is characterized by mixed bone resorption and formation. METHODS: Immunohistopathology of DSO in the clinically acute and subacute phases was compared with healthy bone. RESULTS: Receptor activator of nuclear factor kappaB ligand (RANKL) was found in DSO lesions. When it was used in vitro to stimulate monocytes, cathepsin K expression was observed in mononuclear prefusion precursors and in multinuclear giant cells. Similarly, exacerbations of DSO were characterized by RANKL and induction of cathepsin K in mononuclear precursor cells, which subsequently seem to differentiate into osteoclasts or foreign body giant cells. The proportion of bone to soft tissue increased with the duration of disease. CONCLUSIONS: RANKL-driven osteoclastogenesis and acidic cysteine endoproteinase cathepsin K seem to play important roles in DSO as osteoclast-mediated bone resorption may represent the primary disease process later followed by new bone formation.

Degradation kinetics of fluorouracil-acetic-acid-dextran conjugate in aqueous solution.

The degradation kinetics of fluorouracil-acetic-acid-dextran conjugate (FUAC-dextran) was investigated in various buffer solutions with different pH value and physiological saline solution at 60 degrees C and 37 degrees C, respectively. The hydrolytic reaction displayed pseudo-first-order degradation kinetics. Hydrolytic rate constant obtained was the function of pH value and independent of species of buffering agents. The smallest rate constant was observed at pH round 3.00. The activation energy of the hydrolytic reaction was estimated from Arrhenius equation as 88.73 +/- 6.00 kJ.mol-1. The special base catalytic degradation of the conjugate was observed from acidic to slight alkaline condition and the special base catalytic rate constants were calculated. The conjugate was more stable in physiological saline than that in buffer solution at pH 7.00 or 9.00 at 37 degrees C. The results revealed that the conjugate was stable in acidic condition and will degrade in alkaline condition.

Pannus invasion and cartilage degradation in rheumatoid arthritis: involvement of MMP-3 and interleukin-1beta.

OBJECTIVE: Synovial inflammation in rheumatoid arthritis (RA) leads to pannus tissue invasion and destruction of cartilage/bone matrix by proteinases. Our intention was to analyze some of the key matrix metalloproteinases (MMPs) in pannus tissue overlying evolving cartilage erosions in RA. METHODS: Frozen tissue samples of pannus and synovium from advanced RA and synovium from osteoarthritic patients were used for immunohistochemical, western blotting and quantitative reverse transcriptase polymerase chain reaction (RT-PCR) analysis of MMP-1, -3, -13 and -14. Synovial fibroblast cultures, stimulated with tumour necrosis factor alpha (TNF-alpha) and interleukin-1 beta (IL-1beta), were analyzed with enzyme-linked immunosorbent assays (ELISA) and quantitative RT-PCR. RESULTS: MMP-3 was highly expressed in pannus tissue compared with significantly lower expression levels of MMP-1, -13 and -14. In fibroblast cultures IL-1beta was a potent stimulus for MMP-3, whereas TNF-alpha was more potent for MMP-1. CONCLUSION: This is the first study to demonstrate quantitatively in real time that MMP-3 mRNA expression is clearly higher in advanced RA pannus tissue compared to parallel RA or osteoarthritic synovium. MMP-3 mRNA levels were also clearly overexpressed in RA pannus compared to MMP-1, -13 and -14. Advanced RA has previously been found to overexpress IL-1beta. The high expression of MMP-3 in pannus and IL-1beta, mediated stimulation of MMP-3 suggest that MMP-3 plays a significant role in the progression of erosions through the proteoglycan-rich cartilage matrix.