A new method: Characterize and quantify biofilm wrinkles by UNet and Sholl Analysis.

The wrinkles on the biofilm contain a lot of information about biofilm growth, so it is essential to characterize and quantify these wrinkles from the original microscopic images to discover more rules governing the biofilm morphology evolution. However, the existing methods to extract the wrinkles are time-consuming, error-prone, and require manual calibration. We propose a new system: using a deep learning method - UNet to identify the biofilm wrinkles in the original experimental images, which can achieve fast and accurate extraction of wrinkles on biofilms. Combining the result of UNet and medical neuron analysis method - Sholl Analysis, we can easily characterize and quantity the B. subtilis biofilm wrinkles. We proposed new characterization parameters such as wrinkle density, wrinkle length, and wrinkle projection area, which can precisely partition the biofilm surface wrinkles into different regions from the biofilm center to the edge, different regions correspond to different growth stages. Our system can be applied to study biofilms growing in different kinds of environments and to study the biofilm growth mechanisms.

Magnetic and near-infrared-II fluorescence Au-Gd nanoclusters for imaging-guided sensitization of tumor radiotherapy.

The significant role of multifunctional nanoprobes with complementary advantages in magnetic and near-infrared-II (NIR-II, 1000-1700 nm) fluorescence properties has been documented in precision cancer theranostics. However, certain limitations, including the large size (>10 nm), low NIR-II fluorescence quantum yield (QY < 1.0%), and inefficient magnetic performance (relaxation rate < 5.0 s-1 mM-1) of nanoprobes, restrict their biomedical applications and clinical translation. Albumin-based biomineralization was adopted to prepare bright NIR-II Au NCs, which were further conjugated with DTPA and Gd ions to produce magnetic and NIR-II Au-Gd NCs. Albumin-based biomineralization helped to develop ultrasmall Au-Gd nanoclusters with ultrasmall size (∼2 nm), high NIR-II fluorescence QY (∼3.0%), and effective magnetic resonance imaging (MRI) performance (relaxation rate (r1) = 22.6 s-1 mM-1). On the one hand, Au-Gd NCs achieved NIR-II fluorescence and MRI dual-modal imaging of tumors with a high signal-to-background ratio (SBR = 8.2) in mice. On the other hand, their effective metabolism simultaneously through the kidney and liver minimized their toxicity in vivo. Moreover, compared to the control group, the survival time of tumor-bearing mice was extended by three times when Au-Gd NCs with high-Z elements were used to perform dual-modal imaging-guided sensitization of tumor radiotherapy. Thus, ultrasmall nanoprobes with complementary imaging modalities and therapeutic functions manifest great potential in cancer precision diagnosis and therapy.

Three-year observations on the effect of different cusp inclinations on the restoration of short maxillary first molar implants: A randomized controlled trial.

Objective: To investigate the effect of different cusp inclination on short implant prosthesis of maxillary first molar after 3 years of weight-bearing in biology and mechanics. Methods: The clinical patients were randomly selected from the database and divided into four groups A, B, C, and D according to the cusp inclination of the maxillary first molar short implant restoration (4.8 mm × 8 mm, Dentium). 20 cases in each group. The cusp inclination was 10 degrees-15 degrees, 15 degrees-20 degrees, 20 degrees-25 degrees, 25 degrees-30 degrees. After 3 years of weight-bearing, cone beam computed tomography (CBCT) and Florida probe were used to measure and observe the height of alveolar bone (H), periodontal probing depth (PD) and modified sulcus bleeding index (MBI). Visual analogue scale (VAS) was used to evaluate the overall satisfaction of patients, and the mechanical complications of each group within 3 years of implant weight-bearing were counted. Results: The H and PD of group D were 1.09 ± 0.23 and 2.19 ± 0.11 respectively, which were significantly higher than those of group A, B and C (p < 0.05). There was no significant difference in MBI between groups A-D (p > 0.05). The VAS scores of group B and group C were 88.36 ± 5.12 and 88.70 ± 4.52 respectively, which were higher than those of group A and group D (p < 0.05). The incidence of food impaction, porcelain collapse and abutment loosening in group D were 40.0%, 25.0% and 15.0% respectively, which were higher than those in group B and C (p < 0.05). Conclusion: The risk of biological and mechanical complications increases after long-term weight-bearing of maxillary first molar short implant prostheses with high cusp inclination. The cusp inclination of short implant prostheses should be designed as low as 25 degrees.

Development and Validation of a Deep Learning Algorithm to Automatic Detection of Pituitary Microadenoma From MRI.

Background: It is often difficult to diagnose pituitary microadenoma (PM) by MRI alone, due to its relatively small size, variable anatomical structure, complex clinical symptoms, and signs among individuals. We develop and validate a deep learning -based system to diagnose PM from MRI. Methods: A total of 11,935 infertility participants were initially recruited for this project. After applying the exclusion criteria, 1,520 participants (556 PM patients and 964 controls subjects) were included for further stratified into 3 non-overlapping cohorts. The data used for the training set were derived from a retrospective study, and in the validation dataset, prospective temporal and geographical validation set were adopted. A total of 780 participants were used for training, 195 participants for testing, and 545 participants were used to validate the diagnosis performance. The PM-computer-aided diagnosis (PM-CAD) system consists of two parts: pituitary region detection and PM diagnosis. The diagnosis performance of the PM-CAD system was measured using the receiver operating characteristics (ROC) curve and area under the ROC curve (AUC), calibration curve, accuracy, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and F1-score. Results: Pituitary microadenoma-computer-aided diagnosis system showed 94.36% diagnostic accuracy and 98.13% AUC score in the testing dataset. We confirm the robustness and generalization of our PM-CAD system, the diagnostic accuracy in the internal dataset was 96.50% and in the external dataset was 92.26 and 92.36%, the AUC was 95.5, 94.7, and 93.7%, respectively. In human-computer competition, the diagnosis performance of our PM-CAD system was comparable to radiologists with >10 years of professional expertise (diagnosis accuracy of 94.0% vs. 95.0%, AUC of 95.6% vs. 95.0%). For the misdiagnosis cases from radiologists, our system showed a 100% accurate diagnosis. A browser-based software was designed to assist the PM diagnosis. Conclusions: This is the first report showing that the PM-CAD system is a viable tool for detecting PM. Our results suggest that the PM-CAD system is applicable to radiology departments, especially in primary health care institutions.

Integrative Molecular Analyses of an Individual Transcription Factor-Based Genomic Model for Lung Cancer Prognosis.

OBJECTIVE: Precision medicine with molecular profiles has revolutionized the management of lung cancer contributing to improved prognosis. Herein, we aimed to uncover the gene expression profiling of transcription factors (TFs) in lung cancer as well as to develop a TF-based genomic model. METHODS: We retrospectively curated lung cancer patients from public databases. Through comparing mRNA expression profiling between lung cancer and normal specimens, specific TFs were determined. Thereafter, a TF genomic model was developed with univariate Cox regression and stepwise multivariable Cox analyses, which was verified through the GSE72094 dataset. Gene set enrichment analyses (GSEA) were presented. Downstream targets of TFs were predicted with ChEA, JASPAR, and MotifMap projects, and their biological significance was investigated through the clusterProfiler algorithm. RESULTS: In the TCGA cohort, we proposed a TF-based genomic model, comprised of SATB2, HLF, and NPAS2. Lung cancer individuals were remarkably stratified into high- and low-risk groups. Survival analyses uncovered that high-risk populations presented unfavorable survival outcomes. ROCs confirmed the excellent predictive potency in patients' prognosis. Additionally, this model was an independent prognostic indicator in accordance with multivariate analyses. The clinical implication of the model was well verified in an independent dataset. High risk score was in relation to carcinogenic pathways. Downstream targets were characterized by immune and carcinogenic activation. CONCLUSION: The proposed TF genomic model acts as a promising marker for estimation of lung cancer patients' outcomes. Prospective research is required for testing the clinical utility of the model in individualized management of lung cancer.

Comparison between computed tomography and magnetic resonance imaging in clinical diagnosis and treatment of tibial platform fractures.

BACKGROUND: Tibial plateau fracture is one of the common fracture types. It occurs mainly in teenagers and is usually caused by a fall. After the occurrence of fracture, knee swelling, pain, limited activity, etc. greatly affect the patient's exercise habits and lifestyle. X-ray, computed tomography (CT) and magnetic resonance imaging (MRI) were used in this examination. X-rays are relatively new and easy to operate. However, there are some errors in the observation of fracture collapse and fracture displacement. In recent years, CT and MRI have been actively used to diagnose various types of clinical fractures. They have more diagnostic power than X-ray film. However, some scholars believe that CT is also prone to errors in clinical application. The volume effect leads to missed diagnosis and misdiagnosis in some cases, while the multidirection scanning of MRI technology can effectively overcome the shortcomings of CT. To facilitate the selection of clinical examination regimens, this study further observed the diagnostic ability of these two regimens in the diagnosis of tibial plateau fractures. AIM: To explore the value of nuclear MRI and CT in the clinical diagnosis of tibial plateau fractures. METHODS: A total of 120 patients with tibial plateau fractures admitted from September 2017 to August 2019 were included. All patients were examined by nuclear MRI and CT scanning. The results were sent to senior physicians in our hospital to complete the diagnosis. RESULTS: Nuclear magnetic resonance showed the same effects as CT in four aspects: fracture displacement, bone defect, fracture site and fracture comminution. There was no significant difference in the score data (P > 0.05). Nuclear magnetic resonance and CT tended to be consistent in the B3, C2 and C3 fracture diagnosis coincidence rate, combined injury detection rate and fracture detection rate. The diagnostic coincidence rate of type B1, B2 and C1 fractures and the accuracy rate of overall fracture classification indicated that the MRI technique was significantly better than that of CT (P > 0.05). CONCLUSION: MRI and CT have good diagnostic typing in the diagnosis of tibial plateau fractures, but MRI is more accurate and may be preferred.

Orthopedic treatment for open fracture of lower extremities and soft tissue defects in young children and rapid rehabilitation after operation.

Objective: This study aims to explore the clinical features of open fracture of lower extremities with soft tissue injury in young children, and its surgical and postoperative rehabilitation methods.Methods: A total of 32 children with open fracture of lower extremities and soft tissue injury treated in our department from January 2010 to December 2016 were included into this study. An individualized treatment plan was established according to the characteristics of each patient's condition, and rapid rehabilitation procedures were carried out after the operation to promote the healing of the wound, and shape and function recovery.Results: In the present study, four children developed superficial infection after the operation, and healed after proper management. Furthermore, two children were found to have osteomyelitis during follow-up after discharge, and healed after they returned to the hospital for debridement. All patients were followed up until fracture healing, and the follow-up duration ranged within 6-15 months, with an average duration of 8.5 months. All patients achieved bony union, and none of the patients had bone exposure. In the curative effect assessment, 24 children were classified as excellent, six children were classified as good, one child was classified as moderate, and one child was classified as poor. The excellent and good rate was 93.75%.Conclusions: Open fracture of the lower extremities commonly occur in the middle and lower segment of the tibia and fibula in children, and is usually combined with soft tissue injury. The correct treatment of fractures, and skin and soft tissue injuries, as well as the rapid rehabilitation of children, are the keys to achieving a good clinical effect.

Effects of postoperative enteral nutrition combined with adjuvant radiotherapy on inflammatory response, nutrition, healing and prognosis in patients receiving radical surgery for esophageal carcinoma.

PURPOSE: To explore the influence of postoperative enteral nutrition combined with adjuvant radiotherapy on inflammatory response, nutrition, healing and prognosis in patients undergoing radical surgery for esophageal carcinoma. METHODS: A total of 114 patients with esophageal carcinoma receiving radical surgery from January 2016 to July 2017 composed the observation group and randomly divided into control group (n=57) and study group (n=57). Patients in the control group were given routine nutritional support after surgery, while those in the study group received enteral nutrition after surgery. The changes in inflammatory response and nutritional level, healing and prognosis in the two groups of patients before and after treatment were compared and analyzed. RESULTS: After treatment, the levels of serum hypersensitive C-reactive protein (hs-CRP) and prostaglandin E (PGE) of patients were decreased in both the control group and study group, and they were lower in the study group than those in the control group, while the levels of serum pro-albumin (PA) and albumin (ALB) of patients in the study group were higher than those in the control group (p<0.05). The postoperative wound healing time, total length of hospital stay, postoperative first exhaust time and defecation time in the study group were shorter than those in the control group (p<0.05). The total incidence rate of postoperative complications of patients in the study group was lower than that in the control group (p<0.05). CONCLUSION: The application of postoperative enteral nutrition combined with adjuvant radiotherapy in patients subjected to radical surgery for esophageal carcinoma can suppress systemic inflammatory response, improve the nutritional condition, promote postoperative wound healing and improve prognosis and therefore it is worthy of promotion in clinical practice.

Effect of preoperative infusion chemotherapy combined with hyperthermia on sPD-L1 and CEA levels and overall survival of elderly patients undergoing radical resection of lung cancer.

PURPOSE: To explore the effect of preoperative infusion chemotherapy combined with hyperthermia on the expressions of sPD-L1 and CEA in elderly patients undergoing radical surgery for lung cancer, and their prognosis. METHODS: 136 elderly patients undergoing radical resection of lung cancer in our hospital from October 2012 to October 2014 were studied. Patients were randomly divided into two groups, namely the combination group and the individual treatment group, with 68 patients in each group. Patients in the individual treatment group received only preoperative chemotherapy, whereas those in combination group received preoperative infusion chemotherapy and preoperative and postoperative hyperthermia. The treatment efficacy, levels of sPD-L1 tumor marker CEA (carcino-embryonic antigen), and T-lymphocyte subsets (CD4+, CD3+, CD8+, CD29+) were compared between the two groups. Three-year follow-up data were collected to compare the overall survival (OS) of the two groups. RESULTS: The effective rates in the combination group and individual treatment group were 87.5 and 67.5%, respectively (p<0.05). After treatment, lower serum levels of CEA and sPD-L1 were seen in the combination group vs the individual treatment group (p=0.036, p=0.008, respectively). Levels of T-lymphocyte subsets CD4+, CD3+, and CD29+ in both groups increased, and were higher in the combination group vs the individual treatment group (p<0.05). Follow-up data demonstrated that OS in the combination group and the individual treatment group was 61.7 and 48.5%, respectively. Significant difference in OS between the two groups was confirmed by Log-rank test (p=0.043). CONCLUSIONS: Preoperative infusion chemotherapy combined with hyperthermia for elderly patients with lung cancer can improve patient immunity, inhibit tumor growth and lengthen overall survival by improving T-lymphocyte subset levels and reducing the circulating tumor cell content.

Seasonal variability of water characteristics in the Challenger Deep observed by four cruises.

Thirty conductivity-temperature-depth profiler casts in the Challenger Deep were conducted during four cruises from December 2015 to February 2017. Two cruises took place in the summer, and two in the winter. The results demonstrated that water characteristics varied seasonally. The temperature minimum values were the same between the four cruises, but its depth was noticeably shallower in the winter than that in the summer. The θ-S diagram indicated that deep water is more saline in the summer than that in winter at the same potential temperature. Mixing is more intense between 5000 and 6800 m in the summer than that in the winter. The dissipation rate and eddy diffusivity vertically averaged between 5000 and 6800 m in the summer were εT = 3.277 × 10-8 m2s-3 and KzT = 2.58 × 10-2 m2s-1, respectively. The geostrophic flows below the reference level of 3000 dbar were cyclonic in the summer, travelling westwards in the northern and eastwards in the southern areas of the Challenger Deep.

Perineal Ultrasound Versus Magnetic Resonance Imaging (MRI) Detection for Evaluation of Pelvic Diaphragm in Resting State.

BACKGROUND The aim of this study was to compare the consistency differences between ultrasound and MRI detection methods and the reliability between 2 independent observers. MATERIAL AND METHODS Under 2 kinds of states - the resting state and muscle contractions state - intra-class correlation coefficients (ICCs) were calculated and the consistency of 2 diagnostic methods was evaluated by 2 independent observers. We also assessed the interscorer reliability of the 2 observers. RESULTS In terms of the evaluation of biological parameters of the pelvic diaphragm, the consistency of the 2 diagnostic methods was moderate. The ICC of pelvic diaphragm area was 0.55 (95% CI 0.35-0.71), anteroposterior diameter was 0.48 (95% CI 0.28-0.64), and transverse diameter was 0.43 (95% CI 0.25-0.63). The ultrasound detection values of the perineal ultrasound were significantly smaller than those of the MRI. In addition, these differences were increased with the rise of the pelvic diaphragm area. CONCLUSIONS By evaluating the pelvic diaphragm in patients with pelvic organ prolapse in the resting state, it was preliminarily confirmed that the consistency of ultrasound and MRI was only moderate. The comparison of these 2 diagnostic methods under the dynamic muscle contraction state needs to be further explored.

Ubiquitin-specific protease 4 improves the prognosis of the patients in esophageal cancer.

Our study mainly investigated ubiquitin-specific protease 4 (USP4) expression in pathogenesis of esophageal cancer. The data showed significantly increased expression of USP4 in cancer tissues compared to that in para-carcinoma tissues (68.38% ± 25.60% vs 13.04% ± 9.95%, P= 0.000) and positive correlation between USP4 and pathology grade (r= 0.249, P= 0.014), although survival analysis revealed that USP4 expression was positively associated with the prognosis (32.4% vs 10.9%, P= 0.043). Grouped analysis revealed that the prognosis of patients with high USP4 expression were significantly better only in the small tumor subgroup (diameter ⩽ 5 cm) (52.6% VS 8.6%, P= 0.001) and the early stage subgroup (stages 1 and 2) (60.0% VS 16.7%, P= 0.006). Moreover, in the subgroup of clinical stages 1 and 2 with tumor diameter ⩽ 5 cm, high USP4 expression prolonged the survival time of esophageal cancer patients more significantly (75.5% VS 5.9%, P= 0.000). Based on these results, we speculated that it was possible to significantly improve the prognosis of patients with low USP4 expression by targeted therapy in early esophageal cancer. Taken together, our study uncovered a previously unknown function of USP4 in esophageal cancer and more investigations would be carried out to further study its regulation gene network and molecular biological mechanism in esophageal cancer.

Prognostic value of microRNA-126 and CRK expression in gastric cancer.

BACKGROUND: MicroRNA (miR)-126, acting as a tumor suppressor, has been reported to inhibit the invasion of gastric cancer cells in part by targeting v-crk sarcoma virus CT10 oncogene homologue (CRK). The aim of this study was to investigate the clinical significance of miR-126/CRK axis in gastric cancer. METHODS: miR-126 and CRK mRNA expression levels were detected by real-time quantitative reverse transcription polymerase chain reaction in 220 self-pairs of gastric cancer and adjacent noncancerous tissues. RESULTS: Expression levels of miR-126 and CRK mRNA in gastric cancer tissues were, respectively, lower and higher than those in adjacent noncancerous tissues (both P<0.001). Low miR-126 expression and high CRK expression, alone or in combination, were all significantly associated with positive lymph node and distant metastases and advanced TNM stage of human gastric cancer (all P<0.05). We also found that the overall survival rates of the patients with low miR-126 expression and high CRK expression were, respectively, shorter than those with high miR-126 expression and low CRK expression. Interestingly, miR-126-low/CRK-high expression was associated with a significantly worse overall survival of all miR-126/CRK groups (P<0.001). Moreover, multivariate analysis identified miR-126 and/or CRK expression as independent prognostic factors for patients with gastric cancer. Notably, the prognostic relevance of miR-126 and/or CRK expression was more obvious in the subgroup of patients with TNM stage IV. CONCLUSION: Dysregulation of miR-126/CRK axis may promote the malignant progression of human gastric cancer. miR-126 and CRK combined expression may serve as an independent predictor of overall survival in patients with advanced gastric cancer.

Prognostic significance of combined microRNA-206 and CyclinD2 in gastric cancer patients after curative surgery: A retrospective cohort study.

AIM: To investigate associations of microRNA (miR)-206 and CyclinD2 (CCND2) expression, alone or in combination, with clinicopathological characteristics and patients' prognosis in gastric cancer. METHODS: MiR-206 and CCND2 mRNA expression levels were detected by real-time quantitative RT-PCR in 220 self-pairs of gastric cancer and adjacent non-cancerous tissues. RESULTS: Compared with the adjacent non-cancerous tissues, the expression levels of miR-206 and CCND2 mRNA were respectively reduced and elevated in gastric cancer tissues dramatically (both P<0.001). Notably, the expression levels of miR-206 in gastric cancer tissues were negatively correlated with those of CCND2 mRNA significantly (r=-0.463, P<0.001). Then, statistical analysis showed that low miR-206 expression and high CCND2 expression, alone or in combination, were all significantly associated with great depth of invasion, positive lymph node and distant metastases, and advanced TNM stage of human gastric cancer (all P<0.05). After that, we also found that the overall survivals of the patients with low miR-206 expression and high CCND2 expression were respectively shorter than those with high miR-206 expression and low CCND2 expression. More interestingly, miR-206-low/CCND2-high expression was associated with a significantly worst overall survival of all miR-206/CCND2 groups (P<0.001). Furthermore, multivariate analysis identified miR-206 and/or CCND2 expression as independent prognostic factors for overall survival in patients with gastric cancer. CONCLUSION: Our data provide evidence that the dysregulation of miR-206-CCND2 axis may contribute to the aggressive progression and poor prognosis of human gastric cancer in clinical settings. Combined detection of their expression might be particularly helpful for surveillance of disease progression and treatment stratification.

Apoptosis-induced anti-tumor effect of Curcuma kwangsiensis polysaccharides against human nasopharyngeal carcinoma cells.

This study aimed to investigate the effect of Curcuma kwangsiensis polysaccharides on the viability of human nasopharyngeal carcinoma cell line CNE-2 cells, and explore the possible mechanisms. CNE-2 cells were treated with various concentrations of C. kwangsiensis polysaccharides, then the proliferation, apoptosis and the protein expression of apoptosis-related regulators p53 and Bcl-2 were assessed. The results demonstrated that C. kwangsiensis polysaccharides can significantly inhibit the proliferation of CNE-2 cells, which was possibly through the induction of apoptosis mediated by attenuating Bcl-2 expression and promoting p53 expression. The present study therefore indicates that C. kwangsiensis polysaccharides could be developed into potential drugs for nasopharyngeal carcinoma treatment.

Prognostic value of ADAM17 in human gastric cancer.

A disintegrin and metalloproteinase-17 (ADAM17, also named as tumor necrosis factor-alpha-converting enzyme) is a member of the ADAM family. Of all ADAMs, the strongest evidence for a role in malignancy exists for ADAM17. Especially, it has been demonstrated that ADAM17 expression was significantly increased in human gastric cancer. The aim of this study was to investigate the association between ADAM17 expression and the clinicopathological features of patients with gastric cancer. The expression of ADAM17 was detected by real-time quantitative RT-PCR in gastric cancer and adjacent non-cancerous tissues. In addition, ADAM17 expression was analyzed by immunohistochemistry in 220 clinicopathologically characterized gastric cancer cases. The expression levels of ADAM17 mRNA and protein in gastric cancer tissues were both significantly higher than those in non-cancerous gastric mucosa. In addition, positive expression of ADAM17 correlated with the degree of tumor differentiation, depth of invasion, lymph node metastases, distant metastases, and TNM stage (all P<0.05). Furthermore, multivariate analysis suggested that lymph node metastases, distant metastases, TNM stage, and ADAM17 expression were independent prognostic indicators for gastric cancer. Our data suggest for the first time that the increased expression of ADAM17 in gastric cancer is associated significantly with aggressive progression and poor prognosis. ADAM17 may be an important molecular marker for predicting the carcinogenesis, progression, and prognosis of gastric cancer.

Surgical treatment of a giant neurofibroma.

Neurofibromatosis type 1, an autosomal dominant inherited disease, presents pathologic symptoms of multiple systems, including neurofibromatosis, skeletal dysplasia, café-au-lait spots in skins, and so on. A 45-year-old man with neurofibromatosis type 1 was reported in this article. The patient presented a giant neurofibroma in his head and neck, dysplasia of skull, facial bones and spinal columns, and multiple café-au-lait spots in systematic skins. Satisfactory curative effects were obtained in this case after tumor resection and prosthesis implantation.

[Establishment and characterization of a nude mice model of human diffuse large B-cell lymphoma].

OBJECTIVE: To establish a diffuse large B-cell lymphoma (DLBCL)-mice model using human DLBCL cell line LY8, to investigate its characteristics of growth and to provide a model for in vivo study of DLBCL pathogenesis and treatment. METHODS: LY8 cells were injected subcutaneously into the right flank of nude mice. Harvested tumor tissues were cut into small pieces of 1.5 mm × 1.5 mm × 1.5 mm and implanted subcutaneously into nude mice. Tumor growth was visualized and the histologic characteristics were documented. Expression of LCA, CD20, CD79α, Ki-67, CD3, CD45RO, bcl-6, MUM-1, CD10 and bcl-2 were examined by using immunohistochemistry. IgH clonal rearrangement and status of three microsatellite loci (D14S68, D18S69, D20S199) in the xenografted tumor samples and the parental cell line LY8 were detected using PCR amplification followed by PAGE. RESULTS: The subcutaneous xenograft DLBCL model was successfully established by using cell line LY8, and a stable growth was achieved up to the 9th generation. The tumor in each generation showed similar growth characteristics and the rate of subcutaneous tumor formation was 91.9% (114/124). The tumor growth was observed from the 2nd week after implantation, reaching 1.3 cm in major diameter at the 3rd week and 2.0 cm at the 4th week. The tumor had identical morphological characteristics with those of human DLBCL, and expressed LCA, CD20, CD79α, bcl-6, MUM-1, CD10 and bcl-2. The tumor of xenograft mice and cell line LY8 showed identical IgH rearrangement and microsatellite length. CONCLUSIONS: A human DLBCL bearing mouse model was successfully established. The mice model is similar to human counterpart with high stability and repeatability. Therefore, it provides an ideal animal model for in vivo studies of the biological characteristics and treatment of DLBCL.

Characterization and evaluation of nanostructured lipid carrier as a vehicle for oral delivery of etoposide.

Nanostructured lipid carriers (NLCs) are a new generation of lipid nanoparticles, which have showed some advantages over traditional lipid nanoparticles, such as improved drug incorporation and release properties. The purpose of this study is to develop an optimized nanostructured lipid carrier formulation for etoposide (VP16), and to estimate the potential of NLCs as oral delivery system. VP16-NLCs were prepared by an emulsification and low-temperature solidification method. The average drug entrapment efficiency, particle size and zeta potential of VP16-NLCs, VP16-PEG40-St-modified NLCs (VP16-PEG40-NLCs) and VP16-DSPE-PE- modified NLCs (VP16-DSPE-NLCs) were 57.9-89.7%, 125.9-91.2 nm and -28.49 to -15.34 mV, respectively. The absorption of VP16-NLCs in the intestine was performed by the diffusion chamber. It was found that VP16-DSPE-NLCs with a smaller particle size made the drug transport easy from mucosal to serosal side. A pharmacokinetic study was conducted in rats. After oral administration of VP16 at a dose of 180 mg/kg in the form of either VP16-NLCs or suspension, the relative bioavailability of VP16-NLCs, VP16-PEG40-NLCs and VP16-DSPE-NLCs were enhanced about 1.8-, 3.0- and 3.5-fold, respectively, compared with VP16 suspension. Furthermore, VP16-DSPE-NLCs displayed the highest cytotoxicity against human epithelial-like lung carcinoma cells. The NLCs formulation remarkably improved the oral bioavailability of VP16 and demonstrated a promising perspective for oral delivery of VP16.

[Study on PI3K inhibitor LY294002 for chemotherapeutic sensitization in diffuse large B cell lymphoma cell lines].

OBJECTIVE: To investigate the effects on chemotherapeutic sensitization of the PI3K inhibitor LY294002 in diffuse large B cell Lymphoma (DLBCL) cell lines ly1, ly8, ly10. METHODS: The three cell lines were treated with LY294002, or doxorubicin alone or combined or sequentially respectively. Western blotting was used to detect the level of phospho-AKT after the treatment. Flow cytometry combined with annexin V-FITC assay and Brdu incorporation assay were used to analyze the alterations of cell cycle, proliferation, and apoptosis, respectively. RESULTS: LY294002 decreased the level of phospha-AKT efficiently in the three DLBCL cell lines. The ratio of S phase cells was significantly decreased (P < 0.05). Sequential use of LY294002 and doxorubicin increased the ratio of apoptosis and there was significant difference between the sequential group and the other four groups (P < 0.05) at 24, 48, 72(ly1), 48, 72 (ly8) or 24 h (ly10). CONCLUSION: LY294002 can sensitize doxorubicin-induced apoptosis and may be a potential molecular therapeutic agent targeted at AKT signaling pathway in DLBCL.