PNAS-4 is a novel pro-apoptosis gene identified latetly. In recent years, there has been a large number of research reports on the basic studies about PNAS-4 in cancer gene therapy and gene therapy of PNAS-4 alone or combined with chemotherapy or radiotherapy manifested a good application prospect, but its molecular mechanisms to promote apoptosis is not clear yet. In this paper, recent research about PNAS-4 in cancer gene therapy is briefly reviewed, and recent hypotheses on its molecular mechanisms to promote apoptosis are especially elucidated. Based on its newly identified characteristics of structural domain, we made a point that PNAS-4 might regulate functions of some target protein related to apoptosis by deSumoylation as a new deSumoylating isopeptidase, and consequently promote apoptosis.
Bone marrow-derived mesenchymal stem cell (BMSC) transplantation is one of the most popular therapeutic measures in severe acute pancreatitis (SAP). However, technical challenges and ethical concern have hindered its clinical application. Paracrine factor, as a new safe and easy handing therapeutic measure, can work comparably effective as BMSC transplantation in SAP therapy, but bio-safe risks could be greatly reduced. In this paper, we reviewed the therapeutic effect and potential mechanism of paracrine factors in the treatment of SAP. The injection of paracrine factors yielded from cultured cell suspension will be a new cell therapeutic measure for SAP.
Objective To compare the healing process and clinical results of bioactive glass and allogenic bone in the repair of bone defects after benign bone tumor curettage. Methods Between November 2011 and December 2012, 20 patients with benign bone tumor received bioactive glass and allogenic bone for repair of bone defects after benign bone tumor curettage. There were 17 males and 3 females, aged 9-68 years (median, 18.5 years). The mean course of disease was 3.3 months (range, 1-9 months). Pathological examination revealed that there were 7 cases of chondroblastoma, 5 cases of bone cyst, 2 cases of non-ossifying fibroma, 2 cases of enchondroma, 1 case of vascular tumor of bone, 1 case of lipoma of bone, 1 case of osteoid osteoma, and 1 case of chondromyxoid fibroma. The lesion located at the femur in 5 cases, at the tibia in 11 cases, at the humerus in 1 case, at the calcaneus in 2 cases, and at the talus in 1 case. The bioactive glass and allogenic cancellous bone were implanted in the cavity at the same time. The Musculoskeletal Tumor Society (MSTS) function evaluation score was used for evaluation of postoperative limb function. According to the imaging and clinical benefit, the healing processes of two kinds of implants were evaluated. The healing rate and healing time were compared. The distribution of the bioactive glass was divided into two layers: the layer close to host bone and the layer close to allogenic bone. The bone ingrowth time and bone resorption time in different layers were evaluated and compared. Results All cases were followed up 12-42 months (mean, 34.5 months). All incisions healed by first intention. There were no complications of wound infection or deep infection, rejection, nonunion of bone, fracture at bone graft site, and collapsing of articular surface. There was no tumor recurrence during follow-up. The mean MSTS functional score was 29.5 (range, 28-30) at last follow-up. Complete healing was observed in 11 cases and healing in 9 cases. The healing rates of two kinds of implants were both 100%. The healing time of bioactive glass and allogenic bone was (4.7±1.3) months and (5.2±1.6) months, respectively, showing no significant difference (t=-1.240, P=0.244). The bone ingrowth time and the bone absorption time were (3.6±0.9) months and (3.7±1.0) months in the layer close to host bone and were (4.2±1.3) months and (4.2±1.3) months in the layer close to allogenic bone, all showing no significant difference (t=1.785, P=0.097; t=1.476, P=0.172). Conclusion For the repair of bone defects after benign bone tumor curettage, bioactive glass can achieve satisfactory healing result and has good safety.
This research is to explore the perfusion time-intensity curve parameters of a lung adenocarcinoma xenograft into nude mouse model with contrast enhanced ultrasonography (CEUS); and to investigate the angiogenesis features of tumor at different growth time. Twenty one lung adenocarcinoma xenografted nude mice were divided into three groups and inculcated with human lung adenocarcinoa. Time window for examining CEUS were respectively in 7-day, 14-day and 28-day. The perfusion parameters including rise time (RT), peak intensity (PI), area under the curve (AUC) of lung tumor were obtained on CEUS images by using off-line software Q lab. Immunohistochemically staining for CD34 was used to observe the microvessel density (MVD).The 7-day group had the highest AUC and PI; AUC and PI of 14-day and 28-day group decreased gradually (P < 0.05). RT was increased as tumor growth. In tumor with necrosis, AUC and PI of non-necrosis part were also larger than necrosis part (P < 0.05). Immunohistochemically staining for CD34 of all tumors reflected that the density of microvessels in necrosis tumor was significantly higher than those without necrosis (7.50±3.44 vs.12.44±5.74, P=0.034). Pearson correlation indicated that PI was positively related with MVD (r=0.668, P=0.008). Lung adenocarcinoma perfusion characteristic can be accessed from time-intensity curve parameters by using noninvasively and non-radiative contrast enhanced ultrasonography. Time-intensity curve parameters including AUC, PI and RT may reflect tumor angiogenesis.
ObjectiveTo explore the influence mechanism of proliferation and invasion in colon cancer cell after silence of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene. MethodsRT-PCR or Western blot method was used to detect the expression of PTEN mRNA or protein among four colon cancer cell lines (HT-29, WiDr, CaCo-2, and Colo320 cell lines). small interfering RNA (siRNA) was used to synthetize PTEN siRNA and transfect it into colon cancer cells. The expression of PTEN protein after transfecting was detected by Western blot. WsT-1 and invasion assay were used to examine the effects of PTEN siRNA silence on proliferation and invasion in colon cancer cells. ResultsPTEN mRNA and protein were expressed in all the four colon cancer cell lines. After PTEN siRNA transfected into the colon cancer cells, the expressions of PTEN proteins were inhibited in all the four colon cancer cell lines (P < 0.01), and the proliferation and invasion of colon cancer cells were enhanced significantly (P < 0.01). ConclusionsPTEN siRNA play an important role in metastasis process of colon cancer via enhanced its proliferation and invasion. Therefore, the understanding biologic mechanisms for regulation of PTEN might enable better molecular target therapy of treating the colon cancer patients with metastasis.
ObjectiveTo investigate regulation mechanism of glypican-3 (GPC3) on Hippo signaling pathway and its effects on biological behavior of hepatocellular carcinoma Huh7 cells. MethodsShort hairpin RNAs (shRNA) targeting GPC3 and Yes-associated protein 1 (YAP1) genes were constructed. All of the shRNAs were transfected into Huh7 cells by liposome transfection in order to screen out the stable expression cell lines. The expressions of GPC3 and YAP1 in Huh7 cells were detected by PCR and Western blot in order to screen out the effective shRNAs. The proliferation, invasion, and apoptosis of Huh7 cells were detected by Edu cell proliferation assay, transwell, and flow cytometry. GPC3 shRNA transfection experiments were divided into 6 groups:non-transfection group, empty vector group, GPC3-714-shRNA group, GPC3-647-shRNA group, GPC3-1718-shRNA group, and GPC3-2134-shRNA group. YAP1 shRNA transfection experiments were divided into 6 groups:non-transfection group, empty vector group, YAP1-906-shRNA group, YAP1-1363-shRNA group, YAP1-1666-shRNA group, and YAP1-2895-shRNA group. GPC3 regulation experiments were divided into 5 groups:non-transfection group, empty vector group, GPC3-1718-shRNA group, GPC3-1718-shRNA+ rhYAP1 group, and YAP1-1666-shRNA group. Results① GPC3-1718-shRNA and YAP1-1666-shRNA plasmids were successfully constructed to silence the expressions of GPC3 and YAP1. ② The expressions of GPC3 mRNA and protein in each transfection group were significantly lower than those in the non-transfection group (P<0.05) and the empty vector group (P<0.05), while which in the GPC3-1718-shRNA group was significantly lower than those in all the other transfection groups (P<0.05). The expressions of YAP1 mRNA and protein in each transfection group were significantly lower than those in the non-transfection group and empty vector group (P<0.05), while which in the YAP1-1666-shRNA group was significantly lower than those in all the other transfection groups (P<0.05). ③ The expressions of YAP1 mRNA and protein in the GPC3-1718-shRNA group and the YAP1-1666-shRNA group were significantly lower than those in the non-transfection group (P<0.05) and the empty vector group (P<0.05), while which in the GPC3-1718-shRNA+rhYAP1 group were significantly higher than those in the GPC3-1718-shRNA group (P<0.05) and the YAP1-1666-shRNA group (P<0.05). ④ Compared with the non-transfection group and the empty vector group, the abilities of cell proliferation and invasion in the GPC3-1718-shRNA group and the YAP1-1666-shRNA group were significantly decreased, and the cell apoptosis was significantly increased (P<0.05); The cell proliferation, invasion, and apoptosis in the GPC3-1718-shRNA+rhYAP1 group were significantly improved (P<0.05). ConclusionGPC3 is likely to affect biological behavior of hepatocellular carcinoma Huh7 cells through regulation of YAP1 in Hippo signaling pathway.
ObjectiveTo explore the dynamic changes of immune cell populations in Panc02 pancreatic cancer bearing immunocompetent mice. MethodsThe C57BL6/J mice syngeneic pancreatic cancer cell line Panc02 were subcutaneously implanted to establish the immunocompetent murine pancreatic cancer bearing model.According to the tumor size, the tumor was classified into 4 stages, named T1-T4, respectively.The flow cytometry was performed to identify the dynamic changes of different cell populations, such as inflammatory cells (CD45+), T helper (Th) lymphocytes, cytotoxic T lymphocytes (CTL), B lymphocytes, granulocytes, macrophages, dendritic cells (DC), natural killer (NK) cells, and natural killer T (NKT) cells in the peripheral blood and tumor tissue. ResultsThree dynamic types of immune cells with the tumor progression were identified:consistent increase, consistent decrease, increase and then decrease.①In peripheral blood:The proportion of the Th lymphocytes, CTL, and B lymphocytes consistently decreased; The proportion of granulocytes consistently increased; The proportion of the DC, macrophages, NK cells, and NKT cells increased from T1 to T3 stage but sharply decreased at T4 stage.②In tumor tissue:The intratumoral CD45+ cells consis-tently increased; The proportion of the granucolyte, macrophages, and DC consistently increased; The proportion of the Th lymphocytes and the CTL consistently decreased; The proportion of the B lymphocytes did not change significantly; The proportion of the NK cells or NKT cells increased from T1 to T3 stage but sharply decreased at T4 stage. ConclusionWith pancreatic cancer progression, the immune cell populations show different dynamic change models, which imply their important roles in predicting the prognosis and the integrated treatments of pancreatic cancer.
ObjectiveTo systematically review quality of evidence-based pneumocystosis guidelines for children, compare the differences and similarities among recommendations, and to provide references for clinical application. MethodsDatabases such as TRIP, PubMed, CNKI, VIP, WanFang Data, CBM, U.S National Guideline Clearinghouse (NGC), and Guidelines International Network (GIN) were searched to collect evidence-based guidelines on medication therapy for children with pneumocystosis. Methodological quality of included guidelines was evaluated according to the AGREE Ⅲ instrument, and the differences and similarities among recommendations were compared. ResultsA total of 3 evidence-based guidelines concerning children with pneumocystosis were included, of which, 2 were made by the USA and 1 by international academic organizations. Only 1 guideline was especially suitable for children, while others were for children of different ages and adults. The results of AGREE Ⅲ scoring showed that, among 6 domains which were rated using the AGREE Ⅲ instrument, "scope and purpose", "stakeholder involvement", "rigor of development", "clarity and presentation" and "editorial independence" were scored more than 60%; while "applicability" was only scored 46%, 25% and 31%. Besides, the grading of evidence and grading of recommendations were different. The medication recommendations of different guidelines were not the same. ConclusionThe quality of 3 guidelines is not high, and the grading of evidence and the strength of recommendation are still needed to be unified. The target population and the recommendations in the guidelines are different.