[Recommendations on the diagnosis and treatment of osteoarthritis in China].

Osteoarthritis (OA) is the most common form of arthritis and the leading cause of old age disability, affecting an estimated 302 million people worldwide. OA is seriously overlooked in the world. The awareness of OA and the popularization of standardized diagnosis and treatment are all lacking. Knees, hips, and hands are the most commonly affected joints in OA. Based on the experience of diagnosis and treatment, consensus and guidelines, we formulated this diagnosis and treatment standard in order to standardize the diagnosis and treatment of OA. We hope that our standard can reduce misdiagnosis and mistreatment and improve the prognosis of OA.

[Relationships between decreased LAMC3 and poor prognosis in ovarian cancer].

Objective: To investigate the correlations of laminin subunit gamma 3 (LAMC3) expression with prognosis of ovarian cancer (OC). Methods: LAMC3 protein expression was measured using immunohistochemical streptavidin-peroxidase-biotin connection method (IHC). Gene expression and related clinical data in the cancer genome atlas (TCGA) cohort and clinical proteomic tumor analysis consortium (CPTAC) were applied to analyse the correlation between gene and protein expressions and clinical outcomes. Correlations between LAMC3 and clinicopathological factors were evaluated using the Pearson χ2 test (2-sided). The probability of survival and significance was calculated using the Kaplan-Meier plot. The functional clustering of biological pathways enriched from co-expressed genes of LAMC3 was used to explore the possible mechanisms that LAMC3 might contribute to poor prognosis. Results: Based on the IHC results of 216 OC tissues or ovaries (including 208 tumors and 8 normal tissues) and 51 OC tissues (including 24 chemotherapy-resistant and 27 sensitive tissues), and the protein expression data from CPTAC (including 100 primary tumors and 25 normal tissues), the results showed that the protein expression of LAMC3 was significantly decreased in OC tissues compared with normal, decreased in advanced-stage tissues compared with early-stage tissues, and decreased in drug-resistant tissues compared with sensitive tissues (all P<0.05). Furthermore, low expression of LAMC3 protein was significantly associated with poor disease-free survival (DFS) and overall survival (OS) in 51 OC tissues (P<0.01), consistent with the results that the low levels of LAMC3 mRNA predicted short DFS and OS in 489 OC tissues of the TCGA cohort (P<0.05). The results suggested that low expression of LAMC3 might be the adverse factors for OC development, such as drug resistance and advanced tumors, and might be a risk indicator for prognosis. Moreover, functional clustering of biological pathways enriched from the co-expressed genes of LAMC3 in TCGA ovarian cohort indicated that LAMC3 potentially involved in regulation of OC via oncogene-pathways such as Ras associated protein 1 (Rap1), mitogen-activated protein kinase (MAPK), Ras and cell adhesion-related pathways such as extra cellular matrix (ECM)-receptor interaction and focal adhesion. It indicated that LAMC3 might contribute to short survival and tumor progression by regulation of the above pathways. Conclusion: Low expression of LAMC3 is related to poor prognosis and malignant progression in OC, and thus it is expected to be a new prognostic marker and therapeutic target for clinical treatment.

Aconitine induces cell apoptosis in human pancreatic cancer via NF-κB signaling pathway.

OBJECTIVE: Pancreatic cancer is one of the leading causes of death from cancer in European countries and the United States. This study sought to investigate the effects of aconitine, a well-known aconitum plant-produced toxin, on pancreatic cancer cell growth and apoptosis and to explore the potential mechanisms. MATERIALS AND METHODS: In this study, pancreatic cancer cell lines Miacapa-2 and PANC-1 were cultured, and cell viability was examined in these two cells treated with different doses of aconitine. Moreover, cell apoptosis was also analyzed upon aconitine treatment, and the specific mechanism was examined by Western blot assay and caspase activity detection. RESULTS: The results showed that aconitine inhibited pancreatic cancer cell growth in a dose- and time-dependent manner. The administration of aconitine in Miapaca-2 and PANC-1 cells also induced cell apoptosis by upregulating the expression of pro-apoptotic factors Bax, cl-caspase-3, cl-caspase-9, and cleaved poly (ADP-ribose) polymerase 1 (PARP1), and by decreasing the anti-apoptotic Bcl-2 expression. More importantly, NF-κB was also decreased upon aconitine treatment. In a xenograft mouse model of pancreatic cancer, aconitine suppressed tumor growth and increased cell apoptosis. CONCLUSIONS: This study is the first report on the effects of aconitine on pancreatic cancer, and it reveals that aconitine may serve as a potent therapeutic strategy for clinical treatment of pancreatic cancer.

Conscious sedation with midazolam and dezocine for diagnostic flexible bronchoscopy.

OBJECTIVE: This study aimed to assess the benefits and risks of conscious sedation with midazolam and dezocine in diagnostic flexible bronchoscopy (FB). PATIENTS AND METHODS: This prospective case control study enrolled 40 non-sedated and 40 sedated subjects who underwent diagnostic FB. All received the standard upper airway preparation, while sedated subjects received midazolam and dezocine for conscious sedation. Subject discomforts during FB were assessed using the verbal analogue score (VAS, 0-10 scale). Willingness to return was assessed as five scales to monitor subject's satisfaction level. Safety profiles throughout the procedures were also assessed. RESULTS: Anterograde amnesia existed in 75.0% sedated subjects. Compared to non-sedated subjects, sedated ones expressed less discomfort, with lower VAS scores regarding scope insertion (4 [0-10] vs. 0 [0-4], p < 0.001), cough (5.5 [0-10] vs. 0 [0-4], p < 0.001), dyspnea (3.5 [0-10] vs. 0 [0-4], p<0.001), pain (3 [0-10] vs. 0 [0-5], p < 0.001), and global tolerance of the procedures (5.5 [1-10] vs. 0 [0-5], p < 0.001). More sedated subjects expressed willingness to return (90.0% vs. 30.0%, p < 0.001). Sedated subjects had no more hypoxemic episodes during the procedure (7.5% vs. 5.0%, p > 0.99), which were all transient and not life-threatening. CONCLUSIONS: Conscious sedation with midazolam and dezocine reduces discomforts, improves satisfaction level, and carries no significantly risks in subjects undergoing diagnostic FB.

Interleukin-27 and interleukin-23 in patients with systemic lupus erythematosus: possible role in lupus nephritis.

OBJECTIVES: To analyse the concentration of interleukin (IL)-27 and IL-23 in serum and urine of patients with systemic lupus erythematosus (SLE) compared with healthy controls (HC). METHOD: An enzyme-linked immunosorbent assay (ELISA) was used to analyse the serum and urine concentration of IL-27 and IL-23 from 50 patients with lupus nephritis (LN), 55 patients without LN, and 30 HC. The correlations between the levels of IL-27, IL-23, and disease activity, clinical parameters in SLE patients were analysed. RESULTS: The levels of IL-27 and IL-23 increased significantly in the serum and urine of SLE patients with and without LN compared with HC. Moreover, urine levels of IL-27 and IL-23 were correlated with the renal SLE Disease Activity Index (rSLEDAI) score and 24-h urinary protein levels. After 6 months of immunosuppressive treatment, urine IL-27 expression rose significantly in SLE patients with LN. CONCLUSIONS: IL-27 and IL-23 may be involved in the pathogenesis of LN.

Profiling of differentially expressed microRNAs (miRNAs) during differentiation of rat hepatic oval cells (HOCs) into hepatocellular carcinoma (HCC) cells.

OBJECTIVE: To explore the expression profile of miRNAs during differentiation of rat hepatic oval cells (HOCs) into hepatocellular carcinoma cells (HCC). METHODS: Proliferation of rat HOCs was induced by chemical carcinogen, 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) in male rats. By using Percoll density gradient centrifugation method, HOCs were isolated, followed by continuous cultivation in vitro. The isolated HOCs were identified via Thy-1 and C-kit detection under laser scanning confocal microscope. Total miRNA was then extracted from HOCs during cell differentiation for microarray hybridization. Differentially expressed miRNAs among the indicated time points were identified. The target genes of identified miRNAs were predicted using PicTar, Target-Scan, and miRanda; then the functions and pathways of the genes were enriched. Y chromosome-specific polymerase chain reaction (PCR) technique was utilized to trace the differentiation of the male HOCs in carcinogen-induced HCC of female rats. RESULTS: It was shown that isolated HOCs expressed stem cells markers of Thy-1 and C-kit in cytoplasm and membrane. Among 1,210 miRNAs identified, 22 were differentially expressed (P < 0.05, fold change ≥2), including 19 up-regulated and 3 down-regulated ones. The predicted target genes of these miRNAs were enriched in several functions, including axon guidance, angiogenesis, post-transcriptional protein modification, and small molecular metabolism. For PCR-based SRY detection, HCC genomic DNA of female rats from the experimental group displayed the same PCR product as that from normal male rat. CONCLUSION: Differentially expressed miRNAs exerted important roles during the differentiation process of HOCs to HCC.

Prenatal ethanol exposure enhances the susceptibility to metabolic syndrome in offspring rats by HPA axis-associated neuroendocrine metabolic programming.

OBJECTIVE: The present study was designed to demonstrate that prenatal ethanol exposure (PEE) could enhance the susceptibility of high-fat diet-induced metabolic syndrome (MS) in adult male offspring via a hypothalamic-pituitary-adrenal (HPA) axis-associated neuroendocrine metabolic programmed mechanism. METHODS: Pregnant Wistar rats were intragastricly administrated ethanol 4 g/kg·d from gestational day 11 until term delivery. All male offspring were fed with high-fat diet after weaning, exposed to an unpredictable chronic stress at postnatal week (PW) 17 and sacrificed at PW20. RESULTS: In PEE group, body weight presented a "catch-up growth" pattern, and the HPA axis exhibited a lower basal activity but an enhanced sensitivity to chronic stress, leading to increased levels of serum glucose, insulin, insulin resistant index, total cholesterol and low-density lipoprotein-cholesterol, and decreased levels of high-density lipoprotein-cholesterol. Furthermore, many lipid droplets and vacuolar degeneration were observed in the hypothalamus, pituitary gland and liver. CONCLUSIONS: PEE induces enhanced susceptibility to MS in adult offspring fed with high-fat diet, and the underlying mechanism involves a HPA axis-associated neuroendocrine metabolic programming alteration.

Livin and caspase-3 expression are negatively correlated in cervical squamous cell cancer.

OBJECTIVE: Overexpression in cancer cells of inhibitor of apoptosis proteins like livin appears to promote tumorigenesis by regulating expression of proteins involved in apoptosis signaling. Here, the authors investigated expression of livin and an apoptosis protein that is known to inhibit, caspase-3, in cervical squamous cell carcinoma. MATERIALS AND METHODS: Their expression was assessed for correlation with tumor invasiveness. Immunohistochemistry for livin and caspase-3 was used in 36 normal cervical tissues and in 98 samples of cervical squamous cell carcinoma. The percentage of cells expressing these proteins was compared between normal and cancer samples. Their expression rates in cancer samples were subsequently compared with one another and with the clinical and pathological characteristics of the samples. RESULTS: Livin was more commonly expressed in tumor samples than in normal tissues, while the opposite pattern was observed for caspase-3. Expression of livin was significantly associated with advanced clinical stage, higher pathological grade, and lymph node metastasis (p < 0.05). Expression of caspase-3 was significantly associated with lower clinical stage, lower pathological grade, and lack of lymph node metastasis (p < 0.05). Finally, expression of livin was negatively correlated to caspase-3 expression in cervical squamous cell carcinoma tissue (r = -0.57, p < 0.05). CONCLUSIONS: Livin may inhibit apoptosis in cervical squamous cell carcinoma by downregulating caspase-3, thereby promoting disease progression.

Gamma-glutamyl transpeptidase level is a novel adverse prognostic indicator in human metastatic colorectal cancer.

AIM: Biomarkers have been utilized for prognosis in colorectal cancer; however, relatively few have been identified. We compared the prognostic value of serum alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and gamma-glutamyl transpeptidase (GGT) with carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) in patients with metastatic colorectal cancer (mCRC). METHOD: Blood samples were collected from 239 patients with mCRC presenting between 2005 and 2010 in the Sun Yat-sen University Cancer Center. RESULTS: CEA (P < 0.001), CA19-9 (P < 0.001), GGT (P < 0.001), ALP (P < 0.001) and LDH (P = 0.001) were statistically significant prognostic factors of overall survival (OS). CEA (P = 0.002) and GGT (P = 0.021) were validated as independent predictors. On univariate analysis, CEA (P = 0.003), CA19-9 (P = 0.006), GGT (P < 0.001) and ALP (P = 0.001) were statistically significant predictive factors of progression-free survival (PFS) in patients having first-line chemotherapy. CEA (P = 0.011) and GGT (P = 0.027) were independent predictors. GGT (P = 0.001), ALP (P = 0.016) and LDH (P = 0.039) levels were correlated with the tumour response rate assessed by CT, whilst CEA (P = 0.724) and CA19-9 (P = 0.822) were not. There was a statistically significant difference in OS (P < 0.001) and PFS (P < 0.001) among patients who had elevations of both CEA and GGT compared with those in whom only one or neither was elevated. CONCLUSION: Among GGT, LDH and ALP, only GGT plays an independent role with CEA in predicting OS and PFS in mCRC. When coupled with CEA, GGT may lead to improved prognostic predictors.

Elevated levels of interleukin-27 and effect on production of interferon-γ and interleukin-17 in patients with Behçet's disease.

OBJECTIVES: To measure the levels of interleukin (IL)-27, interferon (IFN)-γ, and IL-17 in serum and supernatants of stimulated peripheral blood mononuclear cells (PBMCs) of patients with Behçet's disease (BD) compared with healthy subjects, and to evaluate the effect of recombinant human IL-27 (rhIL-27) on IFN-γ and IL-17 production by PBMCs. METHODS: Forty-three patients with BD and 40 healthy controls were included in this study. The serum levels of IL-27, IFN-γ, and IL-17 and their production by PBMCs were measured by enzyme-linked immunosorbent assay (ELISA), together with the effects of levels of rhIL-27 on IFN-γ and IL-17 production by PBMCs. RESULTS: There were significantly higher levels of IL-27, IFN-γ, and IL-17 in sera of patients with BD compared with the controls. Levels of IL-27, IFN-γ, and IL-17 in BD patients with active uveitis were significantly higher than in those without. Moreover, IL-27, IFN-γ, and IL-17 production by stimulated PBMCs was increased in BD patients compared to controls. In the presence of rhIL-27, stimulation of PBMCs from BD patients resulted in decreased production of IL-17 but increased production of IFN-γ. CONCLUSIONS: These findings suggest that IL-27, IFN-γ, and IL-17 play a role in the pathogenesis of BD.

PKB/Akt promotes DSB repair in cancer cells through upregulating Mre11 expression following ionizing radiation.

An elevated DNA-repair capacity in cancer cells leads to radiation resistance and severely limits the efficacy of radiation therapy. Activation of Akt is tightly associated with resistance to radiotherapy, and Mre11 protein has important role during the repair of DNA double-strand breaks (DSBs). In this report, our results showed that inhibition of Akt activity impaired the repair of DSBs in CNE2 cells, whereas activated Akt promoted the repair of DSBs in HeLa cells. Knockdown of Mre11 also impaired the process of DSB repair in both these two cell lines. More importantly, we found that Akt could regulate Mre11 expression. Inhibition of Akt activity by small interfering RNA or LY294002 efficiently downregulated the Mre11 expression in CNE2 cells, and transfection with myr-Akt plasmid in HeLa cells upregulated the Mre11 expression. In addition, luciferase reporter analysis revealed that Mre11 reporter activity increased after transfection with myr-Akt1 plasmids, and this myr-Akt1-induced transcriptional activity was blocked in the presence of LY294002. Further study showed GSK3ß/ß-catenin/LEF-1 pathway was involved in this regulation. Knockdown of ß-catenin or LEF-1 led to the downregulation of Mre11, whereas overexpression of ß-catenin led to upregulation of Mre11. The chromatin immunoprecipitation assay assay showed ß-catenin/LEF-1 heterodimer could directly bind to the promoter of Mre11 in vivo. And the luciferase activity of the pGL3-Mre11 and pGL3-Lef increased in HeLa cells following ß-catenin plasmid co-transfected, but was abolished when the LEF-1-binding conserved sequences of Mre11 promoter were mutated. These results together support Akt can upregulate the expression of Mre11 through GSK3ß/ ß-catenin/LEF pathway to elevate DSB-repair capacity in cancer cells.

The pivotal role of c-Jun NH2-terminal kinase-mediated Beclin 1 expression during anticancer agents-induced autophagy in cancer cells.

The c-Jun NH2-terminal kinase (JNK) pathway represents one subgroup of MAP kinases that are activated primarily by cytokines and exposure to environmental stress. Autophagy is a protein-degradation system characterized by the formation of double-membrane vacuoles termed autophagosomes. Autophagy-related gene beclin 1 plays a key role in autophagosome formation. However, the relationships between activation of JNK pathway, autophagy induction and Beclin 1 expression remain elusive. In this study, we used human cancer cell lines CNE2 and Hep3B to investigate the role of JNK-mediated Beclin 1 expression in ceramide-induced autophagic cell death. Ceramide-treated cells exhibited the characteristics of autophagy (that is, acidic vesicular organelle formation and the LC3-II generation). JNK was activated in these two cell lines exposed to ceramide and the phosphorylation of c-Jun also increased. In the meantime, we found that ceramide upregulated Beclin 1 expression in cancer cells. The upregulation of Beclin 1 expression could be blocked by SP600125 (a specific inhibitor of JNK) or a small interfering RNA (siRNA) directed against JNK1/2 or c-Jun. Chromatin immunoprecipitation and luciferase reporter analysis revealed that c-Jun was involved in the regulation of beclin 1 transcription in response to ceramide treatment. In addition, inhibition of JNK activity by SP600125 could inhibit autophagy induction by ceramide. Furthermore, Beclin 1 knockdown by siRNA also inhibited ceramide-mediated autophagic cell death. JNK-mediated Beclin 1 expression was also observed in topotecan-induced autophagy. These data suggest that activation of JNK pathway can mediate Beclin 1 expression, which plays a key role in autophagic cell death in cancer cells.

[Study on the association between interleukin-5 and eosinophil in nasal polyp].

OBJECTIVE: To explore the expression of IL-5 and its role in the formation and development of nasal polyp. METHOD: 31 patients with nasal polyp, 11 patients with chronic sinusitis(CS) and 6 control cases were chosen. Their IL-5 concentration in tissue homogenate were measured by ELISA. All patients with chronic sinusitis, control cases and 15 patients with nasal polyp were chosen to be counted eosinophils in their HE slice. RESULT: 1. The values of IL-5(pg/ml) in nasal polyp, CS and control group separately were: 23.44 +/- 6.68, 16.41 +/- 3.09, 12.86 +/- 4.17. IL-5 concentration in nasal polyp group was higher than that in the other two groups(P < 0.001). 2. The numbers of eosinophils in nasal polyp, CS and control group were 7.42 +/- 2.33, 1.30 +/- 0.59, 1.07 +/- 0.70 separately, the numbers of eosinophils in nasal polyp group were higher than that in CR or control group (P < 0.05). CONCLUSION: Nasal polyp is a disease characterized by eosinophilia, and IL-5 which activates eosinophil plays part role in the formation of nasal polyp.