Sonic hedgehog signaling pathway induces cell migration and invasion through focal adhesion kinase/AKT signaling-mediated activation of matrix metalloproteinase (MMP)-2 and MMP-9 in liver cancer.

The aberrant activation of sonic hedgehog (SHH) pathway contributes to initiation and progression of various malignancies. However, the roles and underlying mechanisms of SHH signaling pathway in invasion and metastasis of liver cancer have not been well understood. In this study, we found that SHH signaling was activated and correlated with invasion and metastasis in hepatocellular carcinoma (HCC). Enhanced SHH signaling by recombinant human SHH N-terminal peptide (rSHH-N) promoted hepatoma cell adhesion, migration and invasion, whereas blockade of SHH signaling with SHH neutralizing antibody or cyclopamine suppressed hepatoma cell adhesion, migration and invasion. Furthermore, matrix metalloproteinase (MMP)-2 and MMP-9 expressions and activities were upregulated and downregulated by rSHH-N and SHH signaling inhibitor, respectively. The rSHH-N-mediated hepatoma cell migration and invasion was blocked by MMP-specific inhibitors or neutralizing antibodies to MMP-2 and MMP-9. In addition, phosphorylations of AKT and focal adhesion kinase (FAK) were increased and decreased by rSHH-N and SHH signaling inhibitor, respectively. Further investigations showed that activation of AKT and FAK were required for rSHH-N-mediated upregulation of MMP-2 and MMP-9, cell migration and invasion. Finally, we found that SHH protein expression was positively correlated with phosphorylatd FAK Tyr397, phosphorylatd AKT Ser473, MMP-2 and MMP-9 protein expressions in HCC samples. Taken together, our findings suggest that SHH pathway induces cell migration and invasion through FAK/AKT signaling-mediated MMP-2 and MMP-9 production and activation in liver cancer.

Clinicopathologic characteristics and surgical outcomes of hepatocellular carcinoma with portal vein tumor thrombosis.

BACKGROUND: Hepatocellular carcinoma (HCC) has a high predilection for portal vein invasion. Furthermore, the treatment of HCC with portal vein tumor thrombosis (PVTT) is controversial. The objective of this study was to investigate clinicopathologic characteristics and surgical outcomes of HCC patients with PVTT. METHODS: The clinicopathologic data and surgical outcomes of 88 patients HCC with PVTT and 211 patients without PVTT who underwent surgery were retrospectively reviewed. The risk factors and the prognosis of HCC patients with PVTT were determined. RESULTS: Cirrhosis, serum alkaline phosphatase (ALP) > 100 IU/L, tumor size > 8 cm, incomplete tumor capsule, and adjacent organ invasion were risk factors for PVTT in HCC on multivariate analysis. Furthermore, HCC patients with PVTT received more major hepatectomies, had more intraoperative blood loss and greater blood transfusion requirements, and higher incidence of postoperative mortality compared with HCC patients without PVTT. The median overall survival of HCC patients with PVTT after surgery was 9 mo, with the 1-, 2-, and 3-y overall survival rates of 31.1%, 18.3%, and 15.2 %, respectively. AFP level, adjacent organ invasion, and PVTT location predicted overall survival of HCC patients with PVTT. CONCLUSIONS: High serum ALP level, cirrhosis, large tumor, incomplete tumor capsule and adjacent organ invasion are predictors of PVTT in HCC. Surgery is a valid therapy for selected HCC patients with PVTT.

Clinical significance of decreased nidogen-2 expression in the tumor tissue and serum of patients with hepatocellular carcinoma.

BACKGROUND AND OBJECTIVES: Nidogen-2 is a ubiquitous component of basement membrane (BM), which is modified by tumor cells to facilitate tumor invasion. However, the expression and function of nidogen-2 in hepatocellular carcinoma (HCC) remains unknown at present. In this study, we sought to investigate the potential role of nidogen-2 in HCC. METHODS: Nidogen-2 expression in HCC tissues, cell lines, and serum was evaluated by immunohistochemistry, immunoassay, and real-time PCR assays. The regulation of nidogen-2 expression was investigated using doxycycline induction and small interfering RNA analyses. RESULTS: Nidogen-2 was significantly decreased in both HCC tissues and serum (P < 0.001). The decreased expression of nidogen-2 in HCC tissues was significantly correlated with tumor progression factors (P < 0.05). Inhibition of matrix metalloproteinase (MMP)-9 led to significantly upregulate nidogen-2 expression in vitro assays. Moreover, patients with HCC had lowest serum nidogen-2 levels compared with patients with benign liver diseases and normal volunteers. Furthermore, the receiver operating characteristic curve analysis revealed a good diagnostic performance of nidogen-2 for HCC. CONCLUSIONS: These findings suggest that decreased expression of nidogen-2 may have a potential pathogenetic role in the development of HCC and may also have potential diagnostic value for HCC.

Toward Coordinate-based Cognition Dictionaries: A BrainMap and Neurosynth Demo.

Characterizing the functional involvement of specific brain regions has long been a central challenge in cognitive neuroscience. Functional magnetic resonance imaging (fMRI) techniques have offered solutions for mapping functional neural networks. The complex nature of structure-function correspondence makes an elaborate task design difficult to fully capture higher-order cognitive function. Other research practices, such as brain-behavior association or between-group comparisons, are thus widely used to explore cognitive correlations with specific brain regions. However, interpreting the results derived from a specific brain region with their underlying cognitive functions has been too general in publications. Here, we use two examples, i.e., a brain-intelligence correlation study and a depression-control comparison meta-study, to demonstrate use of two neuroimaging online databases, BrainMap and Neurosynth. One key utility of the two databases is collecting results from massive cognitive task-based fMRI (tb-fMRI) studies, i.e., coordinates in standard brain space. Just like looking up a "coordinate-based cognition dictionary", researchers can receive a plethora of related tb-fMRI activation information characterized by cognitive domains, specific cognitive functions, cognitive task paradigms, and related publications. Surprisingly, we found that only less than 1% of brain-behavior association or between-group comparison studies have utilized this dictionary approach. We encourage the community to further engage with the existing databases for specific and comprehensive interpretation of neuroimaging as well as guidance of future experimental tb-fMRI design.

Prognostic significance of serum miRNA-21 expression in human non-small cell lung cancer.

BACKGROUND: Deregulation of microRNAs (miRNAs) plays important roles in tumor progression. The aim of this study was to investigate miR-21 expression in serum of non-small cell lung cancer (NSCLC) and its correlation with prognosis of NSCLC patients. METHODS: Dysregulated miRNAs in NSCLC serum were identified by microarray. MiR-21 expression in NSCLC and control serum was detected by TaqMan RT-PCR assay. The correlation of serum miR-21 with clinicopathological factors of NSCLC patients was analyzed. Furthermore, the prognostic significance of serum miR-21 was analyzed by using Kaplan-Meier curves with log-rank tests and a Cox proportional hazard model. RESULTS: The level of miR-21 expression was higher in NSCLC serum samples than in control serum samples (P < 0.01). High serum miR-21 was significantly correlated with tumor-node metastases stage and lymph node metastasis of NSCLC patients (P = 0.016 and 0.026, respectively). The 3-year actuarial overall survival rates in NSCLC patients with high serum miR-21 expression (39.8%) was significantly shorter than those with low serum miR-21 expression (58.2%; P < 0.001). Furthermore, univariate and multivariate analyses for overall survival showed that serum miR-21 expression was an independent prognostic factor for NSCLC patients (P = 0.015, RR = 2.01, 95% CI: 1.78-3.26). CONCLUSION: Serum miR-21 expression might be useful as a prognostic marker for NSCLC patients.

Chinese Association for the Study of Pain: Expert consensus on diagnosis and treatment for lumbar disc herniation.

Lumbar disc herniation is a common disease in the clinical context and does great harm to either the physical or mental health of patients suffering from this disease. Many guidelines and consensus for the diagnosis and treatment of lumbar disc herniation have been published domestically and internationally. According to the expert consensus, clinicians could adopt tailored and personalized diagnosis and treatment management strategies for lumbar disc herniation patients.

Chinese Association for the Study of Pain: Experts consensus on ultrasound-guided injections for the treatment of spinal pain in China (2020 edition).

Spinal pain (SP) is a common condition that has a major negative impact on a patient's quality of life. Recent developments in ultrasound-guided injections for the treatment of SP are increasingly being used in clinical practice. This clinical expert consensus describes the purpose, significance, implementation methods, indications, contraindications, and techniques of ultrasound-guided injections. This consensus offers a practical reference point for physicians to implement successfully ultrasound-guided injections in the treatment of chronic SP.

Proteomic analysis of anti-tumor effects by Rhizoma Paridis total saponin treatment in HepG2 cells.

Rhizoma Paridis total saponin (RPTS) had been identified as the major components responsible for the anti-tumor effects of the herb Rhizoma Paridis, which had been used in China for centuries to treat many diseases including tumor. To elucidate the anti-tumor mechanism of RPTS, a proteomic analysis was carried out with RPTS treatment in HepG2 cells. More than 50 proteins showed a significant change between control (0.01% DMSO) and RPTS (IC(50) approximately 10microg/ml) treated cells after 48h. Twelve proteins had been identified by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) using peptide fingerprinting from 15 protein spots (density difference >2 fold between the control and RPTS-treated group). Among them, six proteins were down-regulated (dUTPase, hnRNP K, GMP synthase, etc.) and six proteins were up-regulated (DNase gamma, Nucleoside diphosphate kinase A, Centrin-2, etc.) by RPTS treatment in HepG2 cells as determined by spot volume (p<0.05). Most of the identified proteins were associated with tumor initiation, promotion, and progression. These findings might offer valuable insights into the mechanism of anti-tumor effect affected by RPTS treatment in HepG2 cells.

[The safety and outcome of patients with acute myocardial infarction transferred for primary angioplasty].

OBJECTIVE: To evaluate the safety and outcome of patients with acute myocardial infarction (AMI) transferred for primary percutaneous coronary intervention (PCI). METHODS: Data from patients with ST elevation AMI urgently transferred from first admitted hospitals to our cath-lab to receive primary PCI were analyzed. According to time intervals from symptom onset to transfer, the patients were divided into early transfer (< 6 h, n = 26), delayed transfer (6 - 24 h, n = 39) and late transfer (24 h to 1 week, n = 18) group. The major cardiac events during transfer periods and one month after PCI were obtained and echocardiogram and left ventricular systolic functions were compared among groups. RESULTS: There was no serious cardiac event during transfer period and all 83 patients received primary PCI with a mean transfer-to-balloon time about 180 minutes. Success rate of PCI was 92.3% in early transfer group, 89.7% in delayed transfer group, and 94.4% in late transfer group (P > 0.05). At one month follow-up after PCI, 0, 10.3% and 16.7% of patients developed heart failure in early, delayed transfer and late transfer group respectively (P > 0.05 vs. early), the LVEF of early transfer group (53.2% +/- 9.7%) was also significantly higher than delayed transfer group (48.6% +/- 8.2%, P < 0.05) and late transfer group (43.1% +/- 10.3%, P < 0.01). CONCLUSIONS: Transfer patients with AMI for primary PCI is safe in the observed time intervals during acute phase. Early transferred patients are associated with better outcome at 1 month post PCI compared to delayed and late transferred AMI patients.

[Genetic instability detected by flow cytometry: DNA aneuploid and P16 expression in biopsy specimens from lung cancer].

OBJECTIVE: To investigate DNA aneuploid and P16 expression in biopsy specimens from lung cancer, and to study genetic instability and the application of flow cytometry in lung cancer pernicious degree diagnosis. METHODS: Blood cells and cancer cells in biopsy specimens were marked simultaneously with anti-CD45 and anti-P16 fluorescent antibody, and the ratio of CD45+ P16+ cells and CD4- P16+ cells was compared. DNA content in biopsy specimens from lung cancer was detected by flow cytometry. RESULTS: Among the 74 cases of lung cancer, there are 46 cases of DNA aneuploid (62.2%). Thirty-seven cases of lung cancer expressed P16 lowly (50%). Twelve cases of lung cancer only expressed P16 lowly (16.22%), 21 cases of lung cancer only expressed DNA aneuploid (28.38%), and 25 cases not only expressed P16 lowly but also expressed DNA aneuploid (33.78%). Indexes of malign degree, such as P16 low expression or DNA aneuploid could be detected in 58 cases among the 74 cases (78.38%) by flow cytometry. CONCLUSION: P16 low expression and DNA aneuploid are the indexes of lung cancer malign degree, and flow cytometry can be used to study genetic instability and evaluate biopsy specimens from lung cancer.

[Effects of small interfering RNA specific for Her-2 gene on biological behavior of ovarian carcinoma cell].

OBJECTIVE: To explore the effects of small interfering RNA (siRNA) specific for Her-2 gene on biological behavior of ovarian carcinoma cell. METHODS: Her-2 siRNA recombinant plasmid and negative control plasmid were transfected into packing cell line PT67 by liposome, and PT67 was selected by puromycin later. SKOV3 was infected by the virus supernatant of stably transfected PT67 cell lines, and the stably transfected SKOV3 cell lines (SKOV3/siRNA, SKOV3/siRNA-negative) established by selection with puromycin were investigated in terms of the reduction levels of Her-2 mRNA and p185 by RT-PCR and immunohistochemistry. Cell proliferation was assayed with methyl thiazolyl tetrazolium, and cell cycle distribution and cell apoptosis were assayed with flow cytometry. The tumor growth of the null mice was analyzed after injection of SKOV3/siRNA and SKOV3/siRNA-negative into the skin. RESULTS: (1) The stable SKOV3 cell lines with a persistent silence of Her-2 gene were established. (2) The percentages of SKOV3/siRNA in G(0)/G(1) phase and S phase were 68.6%, 15.1% respectively; while the percentages of SKOV3/siRNA-negative in G(0)/G(1) phase and S phase were 55.8%, 23.3%. (3) The percentage of SKOV3/siRNA in early apoptosis was (10.500 +/- 0.250)%, while the percentage of SKOV3/siRNA-negative was (0.340 +/- 0.010)% (P < 0.01). (4) Compared with SKOV3/siRNA-negative, the proliferation of SKOV3/siRNA was delayed obviously (P < 0.05), and the growth of the corresponding implanted tumor slowed down significantly (P < 0.01). CONCLUSION: siRNA can inhibit the expression of Her-2 gene effectively, which restrains the biological behavior of ovarian carcinoma cell.

[A clinical comparative study on treatment of severe newly diagnosed immune thrombocytopenia by recombinant human thrombopoietin combined with glucocorticoid].

OBJECTIVE: To evaluate the efficacy and safety of recombinant human thrombopoietin (rhTPO) combined with glucocorticoid in treatment of severe newly diagnosed primary immune thrombocytopenia (ITP). METHODS: From June 2009 to December 2012, 24 male patients and 38 female patients with the diagnosis of severe primary ITP in our hospital were randomized into trial group (31 cases) or control group (31 cases), the median age was 50 years (range: 21-84 years). Trial group was treated with rhTPO combined with glucocorticoid, and control group was treated with glucocorticoid only. RESULTS: At the day 3, 7 and 14 from the beginning of treatment, the average platelet count (APC) in trial group[(35.5±24.9)×109/L, (135.2±94.9)×109/L and (192.0±109.1)×109/L]were significantly higher than that in control group[(24.5±15.6)×109/L, (78.2±121.9)×109/L and (95.8±60.5)×109/L, P=0.022, 0.009 and 0.001, respectively]. There was no significant difference in APC between the two groups at day 28 and 90 after treatment[(147.8±59.1)×109/L vs (105.1±56.9)×109/L, P=0.243; (137.4±52.3)×109/L vs (104.3±59.8)×109/L, P=0.568, respectively]. At the day 7, 14 and 28, the complete response rates in trial group were 61.3%, 87.1% and 80.6%, which were also significantly higher than that in control group (16.1%, 29.0% and 48.3%, P=0.000, 0.000 and 0.004, respectively). The median time to response in trial group was 3 days while in the control group was 5 days; the median duration of complete response in trial group was 76 days while in the control group was 54 days. In trial group, there were 4 cases treated with platelet transfusion, while in control group there were 11 cases, respectively. CONCLUSION: For patients with severe primary ITP, rhTPO combined with glucocorticoid could rapidly increase the platelet count, significantly improve the complete response rate and prolonged the effect with a low incidence of tolerable adverse events compared to single use of glucocorticoid. rhTPO combined with glucocorticoid could be a new therapeutic choice to those patients.

[Evaluation of the impact of R-CHOP chemotherapy on efficacy, safety and prognosis in newly diagnosed diffuse large B-cell lymphoma patients and its prognostic impact: a multicenter retrospective study with long term follow-up].

OBJECTIVE: To evaluate the efficacy, safety and prognostic impact of rituximab plus CHOP (R-CHOP) regimen on patients with diffuse large B-cell lymphoma (DLBCL), to access the impact of R-CHOP on patients' prognosis and to compare that with CHOP regimen. METHODS: Five hundred and seven newly diagnosed DLBCL patients were enrolled from Jan. 1, 2000 to May 1, 2010. Patients were administered with 6 cycles of CHOP or at least 4 cycles of R-CHOP treatments. Rituximab was administered intravenously on day 1 at a dose of 375 mg/m(2). The typical CHOP regimen include cyclophosphamide (750 mg/m(2), IV), doxorubicin (50 mg/m(2), IV) and vincristine (1.4 mg/m(2), IV, maximum 2 mg) and prednisone (60 - 100 mg, oral, day 3 - 7). The complete response (CR) rates, overall response (OR) rates, and side events of these 2 groups were compared. RESULTS: Of the 411 analyzable patients, 224 received CHOP regimen and 187 received R-CHOP regimen. CR rate for R-CHOP group and CHOP group was 77.01% and 71.43%, respectively. OR rate in R-CHOP group was higher than that in the CHOP group (95.19% vs 87.95%, P = 0.007). The median follow-up time of R-CHOP group was 28.1 months vs that of 35.2 months in CHOP group. There was significant difference in progression free survival (PFS) and overall survival (OS) between 2 groups (P = 0.018 and 0.034, respectively). At the end of follow-up, the estimated median PFS in R-CHOP group had not been reached, while that was 84.8 months in CHOP group. The median OS in both groups had not yet been reached. The adverse events in R-CHOP group were similar with that in CHOP group. CONCLUSIONS: R-CHOP is a safe and effective regimen for management of newly diagnosed DLBCL, with a better remission rate, PFS and OS.

MicroRNA-21 modulates chemosensitivity of breast cancer cells to doxorubicin by targeting PTEN.

BACKGROUND AND AIMS: Ovexpression of microRNA-21 (miR-21) is found in various human cancers. Our aim is to investigate the association of miR-21 expression with the sensitivity of breast cancer cells to doxorubicin (ADR). METHODS: The half maximal inhibitory concentration (IC(50)) value of ADR in resistant MCF-7/ADR or parental MCF-7 cells was determined by MTT assay. TaqMan RT-PCR or Western blot assay was performed to detect the expression of mature miR-21 and tumor suppressor gene (PTEN) protein. MCF-7 or MCF-7/ADR cell line was transfected with miR-21mimic or inhibitor. The IC(50) value of ADR was determined. Flow cytometry and TUNEL assays were performed to analyze apoptosis. The activity of caspase-3 was analyzed. RESULTS: The IC(50) of ADR in MCF-7 and MCF-7/ADR cells was 0.21 ± 0.05 and 16.5 ± 0.08 µmol/L, respectively. We showed that upregulation of miR-21 in MCF-7/ADR cells was concurrent with downregulation of PTEN protein. MiR-21 mimic or inhibitor could obviously affect the sensitivity of breast cancer cells to ADR. Moreover, miR-21 inhibitor could enhance caspase-3-dependent apoptosis in MCF-7/ADR cells. Overexpression of PTEN could mimic the same effects of miR-21 inhibitor in MCF-7/ADR cells and PTEN-siRNA could increase the resistance of MCF-7 cells to ADR. MiR-21 inhibitor could increase PTEN protein expression and the luciferase activity of a PTEN 3' untranslated region-based reporter construct in MCF-7/ADR cells. PTEN-siRNA could partially reverse the increased chemosensitivity of MCF-7/ADR cells induced by miR-21 inhibitor. CONCLUSIONS: Dysregulation of miR-21 plays critical roles in the ADR resistance of breast cancer, at least in part via targeting PTEN.

[Construction of recombinant deltaN IkappaBalpha adenovirus and its suppression of NF-kappaB activity in A549 cells].

AIM: To explore the regulatory effect of deltaN IkappaBalpha gene on the activity of nuclear factor-kappaB(NF-kappaB). METHODS: Ser32-and Ser36-deleted IkappaBalpha gene (deltaN IkappaBalpha) was cloned into adenovirus vector, and a replication-defective recombinant deltaN IkappaBalpha adenovirus(Ad-deltaN IkappaBalpha) was generated. A549 cells were divided into three groups: LPS-stimulated groups, Ad-LacZ+LPS group and Ad-deltaN IkappaBalpha+LPS group. Ad-LacZ+LPS group and Ad-deltaN IkappaBalpha+LPS group were infected with Ad-LacZ and Ad-deltaN IkappaBalpha, respectively, two days before LPS stimulation. The NF-kappaB activity of A549 cells was detected by Western blot and electrophoretic mobility shift assay (EMSA). TNF-alpha and IL-6 in the culture supernatant were detecteded by ELISA. RESULTS: The activity of NF-kappaB and the levels of TNF-alpha and IL-6 from Ad-deltaN IkappaBalpha virus-infected A549 cells were significantly decreased as compared with that of LPS-stimulated group and Ad-LacZ+LPS group. CONCLUSION: The results indicated that deltaN IkappaBalpha may inhibit the activation of NF-kappaB and reduce the release of TNF-alpha and IL-6, suggesting the recombinant deltaN IkappaBalpha adenovirus may be used for anti-inflammatory therapy.

[Prokaryotic expression of endostatin and preparation of polyclonal antibody].

BACKGROUND & OBJECTIVE: The aim of this study was to express and purify human endostatin and to prepare polyclonal antibody of mouse anti-human endostatin. METHODS: The cDNA of endostatin was amplified by PCR, then recombined into prokaryotic expression vector and transformed into Escherichia coli BL21 for expression; the mice were immunized with purified products. RESULTS: Prokaryotic expression vector pQE-30 of human endostatin was successfully constructed; the expression product was gained after pQE-30 was transferred into BL21. After purified by Ni affinity chromatography, the product was identified to be a single component by SDS-PAGE. Western blot analysis showed that high titer mouse anti-human endostatin polyclonal antibody was successfully prepared. CONCLUSION: Highly purified expression product and prepared polyclonal antibody provide the necessary material for further study.