Cannabidiol regulates apoptosis and autophagy in inflammation and cancer: A review.

Cannabidiol (CBD) is a terpenoid naturally found in plants. The purified compound is used in the treatment of mental disorders because of its antidepressive, anxiolytic, and antiepileptic effects. CBD can affect the regulation of several pathophysiologic processes, including autophagy, cytokine secretion, apoptosis, and innate and adaptive immune responses. However, several authors have reported contradictory findings concerning the magnitude and direction of CBD-mediated effects. For example, CBD treatment can increase, decrease, or have no significant effect on autophagy and apoptosis. These variable results can be attributed to the differences in the biological models, cell types, and CBD concentration used in these studies. This review focuses on the mechanism of regulation of autophagy and apoptosis in inflammatory response and cancer by CBD. Further, we broadly elaborated on the prospects of using CBD as an anti-inflammatory agent and in cancer therapy in the future.

Global Research Trends in Dental Stem Cells: A Bibliometric and Visualized Study.

Dental stem cells (DSCs) are mesenchymal stem cell-like populations with self-renewal and multidifferentiation potential. These cells have been studied in regenerative medicine and tissue engineering. Despite rapid progress in the past two decades, there has been no bibliometric analysis of DSC research. Here, we performed a comparative study using bibliometric methods for DSCs. A total of 5498 articles were included. Our results showed that the United States was the leader in international cooperation and numbers of citations and was the largest contributor. The Journal of Endodontics published the largest number of articles. The author with the greatest contribution was Songtao Shi. The keywords were mainly related to the fields of tissue engineering and regenerative medicine. Relative research interest and the number of publications increased yearly worldwide. The hotspots of DSC research were transiting from basic research to clinical regenerative medicine. Impact statement Dental stem cells (DSCs) are stem cells with self-renewal and multidirectional differentiation potential. Research in this field is attracting increasing attention. This study aimed to understand the current research status of DSCs and to predict promising keywords and trends. We found the global trend of DSCs and their application in the field of regenerative medicine and tissue engineering. Our study makes a significant contribution to the literature because it will help researchers to understand the research trends and directions in this field.

Use of two kinds of antioxidants to restore the bond strength of bleached enamel.

OBJECTIVES: This work aimed to evaluate the ability of two kinds of antioxidants, namely, grape-seed extract and sodium ascorbate, in restoring bond strength at the resin-enamel interface after bleaching. METHODS: Ten groups of samples with 15 teeth per group were prepared for shear-bond-strength test at the resin-enamel interface after bleaching. The groups were as follows: control; no antioxidant; 2.5%, 5%, 10%, or 15% grape-seed extract; and 2.5%, 5%, 10%, or 15% sodium ascorbate. The peak values of shear bond strength when resin was debonded from teeth and the failure modes under a microscope were recorded. Ten other groups of teeth with two teeth per group were prepared and treated in a similar approach before resin bonding. The samples were cut vertically to the bonding interface. The structures of the bonding interface were compared by scanning electron microscopy. RESULTS: No statistically significant difference in shear bond strength was found among the no-antioxidant, 2.5% grape-seed extract, and 2.5%, 5%, or 10% sodium ascorbate groups (P>0.05), which were statistically significantly lower than the control group (P<0.05). Evidence of marginal gap was observed at the resin-enamel interface, and resin tags in enamel were short, poorly defined, and fragmented. No statistically significant difference in shear bond strength was found among 5%, 10%, or 15% grape-seed extract, 15% sodium ascorbate, and control groups (P>0.05). No evidence of discontinuity was found at the adhesion interface, and resin tags in enamel were long, well defined, and structurally intact. Failure in the adhesive joint was the major debond mode in all experimental groups. CONCLUSIONS: Immediately after bleaching, the bond strength of dental enamel significantly decreased. Bond strength can be restored by 5% grape-seed extract or 15% sodium ascorbate in 5 min.

Clinical significance of immunotherapy with combined three kinds of cells for operable colorectal cancer.

Surgery, chemotherapy, and radiotherapy have presented with the ability of killing tumor cells, as well as damaging the immune function, which can be corrected by the immunotherapy. The purpose of this perspective cohort study was to evaluate the efficacy of postoperative immunotherapies of tumor lysate-loaded dendritic cells (DC), in vitro DC-activated T (DC-AT), and activated T cells (ATC) combined with chemotherapy on the survival of patients with operable colorectal cancer. A total of 253 patients with primary colorectal cancer resection including 181 patients receiving postoperative simple chemotherapy (control group) and 72 patients receiving immunotherapies of DC, DC-AT, and ATC combined with chemotherapy during the corresponding period (immunotherapy group) were enrolled in this perspective cohort study. The survival of these patients was analyzed. The immunotherapy group presented a higher 5-year overall survival rate than the control group (75.63 vs 67.81 %, P = 0.035), as well as 3-year overall survival rate (87.07 vs 74.80 %, P = 0.045). For patients with advanced cancer (TNM stages III and IV), immunotherapy significantly promotes mean survival than control subjects (59.74 ± 3.21 vs 49.99 ± 2.54 years, P = 0.034). Patients who received more than three cycles of immunotherapies had a higher 5-year overall survival rate than those with less than three cycles (82.10 vs 69.90 %, P = 0.035). No serious adverse effect was observed in the immunotherapy group. Postoperative immunotherapies with DC, DC-AT, and ATC combination can promote the survival of patients with operable colorectal cancer (Clinical Trials, ChiCTR-OCH-12002610).

[Assessment value of preoperative neutrophil-lymphocyte ratio for the prognosis of patients with colorectal cancer after radical operation].

OBJECTIVE: To assess the value of preoperative neutrophil-lymphocyte ratio (NLR) for prognosis in patients with colorectal cancer after radical operation. METHODS: Clinical data of 140 patients with colorectal cancer undergoing radical operation in the Department of General Surgery of General Hospital of PLA from July 2005 to July 2011 were analyzed retrospectively. According to preoperative NLR, patients were divided into the low NLR group (NLR<5, n=105) and the high NLR group (NLR≥5, n=35). The overall 5-year survival rates of two groups were compared and the independent risk factors were examined by univariate analysis and Cox model. RESULTS: The overall 5-year survival rates of the low and high NLR groups were 74.8% and 54.7% respectively with significant difference (P=0.03). Univariate analysis revealed depth of tumor, lymph nodes metastasis, TMN stage and NLR were associated with survival (P<0.05, P<0.01). Cox model showed that NLR was independent risk factor of prognosis (RR=1.068, 95%CI:1.009-1.129, P=0.02). CONCLUSION: Preoperative NLR≥5 predicts poorer prognosis of colorectal cancer patients.

[Treatment pattern of adoptive transfer of immune cells and its application in perioperative period for advanced gastric cancer].

Recently immunotherapy for gastrointestinal tumor has rapidly developed, and has improved the effect of cancer comprehensive treatment as an adjunctive therapy in combination with surgery, chemotherapy, and radiation therapy. Adoptive transfer of immune cells is an important treatment method for advanced gastric cancer. In this paper, we reviewed the application of adoptive transfer therapy for advanced gastric cancer in the perioperative period and propose a new model for immunotherapy of advanced gastric cancer based on our experience and the results of clinical experiment.

[Acute ischemic stroke in the very elderly outcome and predictive factors].

OBJECTIVE: To investigate the clinical characteristics, 3-month outcome and predictive factors in the very elderly patients with ischemic stroke. METHODS: A total of 305 acute ischemic patients aged 65 years and over were enrolled in the study. They were divided into two subgroups by age: 80 years old and over (n = 78), 65 - 79 years old (n = 227). The clinical outcome was assessed by the modified Rankin Scale (mRS) on (90 ± 7) days after stroke, and categorized as good (scoring 0 - 2) or poor (scoring 3 - 6) outcome. RESULTS: Significantly lower BMI [(23.62 ± 4.92) kg/m(2) vs (25.08 ± 3.69) kg/m(2), P = 0.005], lower frequency of dyslipidemia (56.41% vs 71.13%, P = 0.006) and alcohol intake (0% vs 6.61%, P = 0.043) were found in the very elderly group. The rates of poor functional outcome in the ≥ 80 years group and the 65 - 79 years old group were 56.41% (44/76) and 41.40% (94/224) respectively, with a P value of 0.015. Multivariate logistic regression analysis showed that higher National Institute of Health stroke scale (NIHSS) total score (OR 1.48, 95%CI 1.19 - 1.83) and lower albumin level (OR 0.73, 95%CI 0.55 - 0.95) were associated with poor outcome in ≥ 80 year old, whereas higher NIHSS total score (OR 1.38, 95%CI 1.24 - 1.53) and complications during hospital stay (OR 2.58, 95%CI 1.07 - 6.19) were predictive factors in the 65 - 79 years old group. CONCLUSION: Our study suggests that NIHSS scores, albumin level and complications during hospitalization are useful predictive factors for the short-term poor functional outcome in the patients of ≥ 65 years old and ≥ 80 years old patients have a worse prognosis.

[Evaluation of in vivo labeled cytotoxic T lymphocytes and cytokine-induced killer cells migration and distribution of subcutaneous gastric tumor model in nude mice].

OBJECTIVE: To investigate the migration and distribution of CTL (cytotoxic T lymphocyte) and CIK (cytokine-induced killer) cells in gastric tumor model. METHODS: Subcutaneous gastric tumor model was established by BGC-823 cancer cells in nude mice. Both CTL and CIK cells were labeled with 99Tc(m) directly and then inoculated into nude mice with subcutaneous tumor by intravenous injection separately. Three mice of each group were evaluated by single-photon emission computerized tomography (SPECT) at 1 h, 6 h and 24 h post-inoculation. After SPECT imaging, 3 mice in each group were sacrificed and got samples of the tumor, liver, spleen, kidney, lung, intestine, etc. The tissue samples were weighed and radioactivity was determined with a well-type scintillation counter. The accumulation of labeled CTL and CIK cells in tissues were expressed as %ID/g (percentage activity of injection dose per gram of tissue) and T/NT (tumor/non-tumor) values were analyzed. RESULTS: The tracing of both cells in SPECT showed a clear migration path away from the injection point to solid tumor, and can be detected in all organs and tissues such as liver, spleen, kidney, lung and intestine, etc not long after injection. The %ID/g peak values of CTL in organs from the highest to the lowest were as follows: tumor (7.79 +/- 0.46), liver (4.12 +/- 0.51), intestine (2.71 +/- 0.16), kidney (1.44 +/- 0.25), spleen (1.24 +/- 0.12), kidney (1.12 +/- 0.11), and all the T/NTs were above 1. The %ID/g peak values of CIK cells in organs from the highest to the lowest were as follows: liver (6.64 +/- 0.67), tumor (5.47 +/- 0.87), intestine (3.55 +/- 0.23), kidney (2.34 +/- 0.41), spleen (1.45 +/- 0.17), lung (1.27 +/- 0.21), and T/NTs > 1 except for liver. After injection, the %ID/g values of tumor in CTL group were 2.35 +/- 0.28 (1 h), 4.58 +/- 0.52 (6 h) and 7.79 +/- 0.46 (24 h) respectively while the %ID/g values of tumor in CIK group 2.23 +/- 0.46 (1 h), 3.25 +/- 0.70 (6 h) and 5.47 +/- 0.87 (24 h) respectively. At 24 h point, the %ID/g of CTL in tumor was much higher than CIK cells (P < 0.05). CONCLUSION: The definite directional tumor-targeting capacity of CTL and CIK cells in tumor-bearing nude mice is promising.

[Experimental study of implantable engineered liver tissue using type I collagen gel as scaffold].

OBJECTIVE: To construct implantable engineered liver tissue (ELT) using type I collagen gel as scaffold. METHODS: Type I collagen was obtained from the tail of a rat. Hepatocytes were collected from a Sprague-Dawley rat, mixed with liquid type I collagen and Dulbecco's modified Eagle's medium to create hepatocyte/collagen gel construct. The construct was inoculated in a 96-well plate. 0, 3, 5, 7, 9, 11, 13, and 15 days after the inoculation the viability of hepatocytes in vitro was measured by MTT assay. Phase contrast microscopy was used to observe the morphology of the hepatocyte/collagen gel construct. Three SD rats underwent injection of the hepatocyte/collagen gel construct into the subcutaneous space. One week later the implant was taken out. The morphology was conducted by routine H.E. staining and immunohistochemical staining. The morphology and function of hepatocytes was investigated by inverted microscopy, routine H.E. staining and immunohistochemical staining. The constructs were also implanted into subcutaneous space, and the differentiation of hepatocytes and the formation of engineered liver tissue were observed by routine H.E. staining and immunohistochemical staining. RESULTS: Phase contrast microscopy showed that the hepatocytes were distributed evenly in the construct and remained round-shape throughout the in vitro culture. MTT assay demonstrated that the high viability of hepatocytes (87%) was maintained up to 7 days, and then decreased gradually. Albumin, the specific marker of hepatocytes remained positive by immunohistochemical staining after 15-day culture. One week after implantation into subcutaneous space, the implanted hepatocytes retained its hepatocyte-specific morphology, i.e. round shape, large nuclear/cytoplasm ratio as well as binuclear cells, and formed small engineered liver tissue containing blood vessels within and surrounding the tissue. CONCLUSION: A novel approach to construct implantable engineered liver tissue using collagen gel as scaffold for growth and differentiation of hepatocytes has been dev eloped. This technique is an attractive tool for the development of liver tissue engineering.

Study of biodegradable and self-expandable PLLA helical biliary stent in vivo and in vitro.

Biodegradable stents have advantages for the treatment of benign and malignant biliary stricture, especially eliminating the need for stent removal. In our present work, helical poly-l-lactic acids (PLLA) stent was fabricated and evaluated in vivo and in vitro. For in vivo study, bile duct injury canine models were made by transection of common bile ducts. Duct to duct anastomosis was done with helical PLLA biodegradable stents. Scanning electron microscopy (SEM) and histopathology were performed after three months. For In vitro study, sludge attachment assessment was performed. Polyethylene (PE) and PLLA membranes were immersed in human bile for two months. The samples were taken out and characterized by SEM. Self-expanding property of the helical stent was tested in 37 degrees Celsius water. The results demonstrate that the biodegradable stent had not only good biocompatibility, but also self-clearing effect to clear the attached sludge away. The self-expanding property facilitated stent implantation and also suggested possibility to be implanted endoscopically.

[In vitro differentiation of human bone marrow-derived mesenchymal stem cells into blood vessel endothelial cells].

OBJECTIVE: To study differentiation of human bone marrow-derived mesenchymal stem cells (BDMSC) into blood vessel endothelial cells for ideal cell origin of complex organ tissue engineering vascularization and injured tissue repairing by cell transplantation. METHOD: After different days of induction with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in 3D fibrin-gels and matrigel, BDMSC and angiogenesis were determined by the utilization of morphological observation, tissue section and CD34, CD31, vascular endothelial growth factor receptor-1 (VEGFR-1, Flt-1), VEGFR-2 (Flk-1), and vWF that were special for blood vessel endothelial cells. RESULT: After 3D-cultured and induced with VEGF and bFGF in vitro in fibrin-gels and matrigel for 3-21 days, BDMSC expressed CD34, CD31, Flt-1, Flk-1, and vWF came into vessel-like configuration. CONCLUSION: VEGF, bFGF as well as Flt-1 and Flk-1, expressed by BDMSC, may form a feasible microenviroment after induction and play an important role during processes of blood vessel endothelial cell differentiation and vessel-like configuration forming of BDMSC. Mesenchymal stem cells may be applied to tissue engineering vascularization and injured tissue repairing by cell transplantation.

[Analysis of the early and late gene expression of lipopolysaccharide activated macrophages by cDNA microarray].

OBJECTIVE: To study the early and late changes in mRNA expression in macrophages in response to lipopolysaccharide (LPS) with a cDNA microarray approach using the Clontech Atlas microarray. METHODS: mRNA was isolated from unstimulated control and LPS stimulated murine peritoneal macrophages at 2 hours and 24 hours poststimulation, converted to (33)P radiolabeled cDNA, and hybridized to mouse array membranes. RESULTS: In macrophages being stimulated for 2 hours, 69 out of 1 176 genes were found to differ by over 3-fold compared with the control. Among them 44 genes were up-regulated and 25 genes were down-regulated. In macrophages stimulated for 24 hours, 11 genes were up-regulated and 26 genes were down-regulated compared with the control. Only 8 genes were identified both at 2 hours and at 24 hours poststimulation. The expressions of many genes encoding transcription factor, cytokines, cell signaling modulators and apoptosis associated proteins were found to have changed. Some genes that were not previously linked to this model, such as bric-a-brac (BTB) and cap-n-collar(CNC) homology 1(BACH1), early growth response protein 2 (EGR2), E47 interaction protein 1 (EIP1), Ngfi-A binding protein 2 (NAB2), myeloblastosis oncogene-like protein (MYBL2), neurofibromatosis 1 (NF1), ciliarry neurotropic factor (CNTF) and semaphorin 4A (Sema4A). CONCLUSION: This study has allowed us to identify genes that may potentially be regulated by LPS at early and late phase in macrophages. These may contribute to better understanding of the mechanism underlying LPS or bacteria induced inflammatory and immune response following infection and trauma.

Relationship between cytokine mRNA expression and organ damage following cecal ligation and puncture.

AIM: To investigate the role of cytokine gene expression in organ damage at different tissue sites during sepsis. METHODS: Male NIH mice were subjected to cecal ligation and puncture (CLP) or sham operation (Sham). Pro-inflammatory cytokine (TNFalpha, IL-1beta and IL-6) and anti-inflammatory cytokine (IL-4) gene expression in the liver and lung tissue were assessed by RT-PCR. The permeability of microvascular and water content in the lungs and liver were also examined. RESULTS: Significant increase in TNFalpha, IL-1beta and IL-6 gene expression was observed at 3 and 12 h after CLP both in the liver and lungs (P<0.01).The level of IL-4 gene expression was not changed after CLP in the lungs, but increased at 12 h after CLP(P<0.01) in the liver tissue. Both the liver and lungs showed a significant increase in microcirculatory permeability at 12 h after CLP(P<0.01), and the increase in the lungs was higher than that in the liver. The water mass fractions in the liver (P<0.05) and lungs (P<0.01) were increased after CLP, and the increase in the lungs happened earlier and more severely than that in the liver. CONCLUSION: The inflammatory response in the liver and lungs was different during sepsis. At the early stage of sepsis, pro-inflammatory reaction dominates both in the liver and lungs. But at the later stage of sepsis, induction of compensatory anti-inflammatory response was seen in the liver but not in the lungs. This difference in situ activity may contribute to the different vulnerability of organ damage during sepsis. The strategy of systemic administration of anti-inflammatory drugs to sepsis should be reconsidered.