[Progress of studies on anti-breast tumor mechanism of alkaloids].

Breast tumor has become one of the malignant tumors with the highest incidence, and is a serious threat to human health, especially to women. Chemotherapy is an important anti-breast tumor therapy, which can be used in almost every stage of breast tumor therapy alone or in the combination with surgery and radiation therapy. Alkaloids are a kind of ubiquitous natural products, and important active components of various medicinal plants. A large number of studies have shown that alkaloids could exert an anti-breast tumor effect by inhibiting proliferation, metastasis and angiogenesis, resisting mitosis, promoting apoptosis and autophagy, and triggering cell cycle arrest. The extensive anti-breast tumor effect makes alkaloids an important candidate drug source. This paper reviews the anti-breast tumor mechanism of natural products of alkaloids.

Circular RNA circ-Foxo3 inhibits esophageal squamous cell cancer progression via the miR-23a/PTEN axis.

Circ-Foxo3 is a circRNA encoded by the human FOXO3 gene and works as a sponge for potential microRNAs (miRNAs) to regulate cancer progression. However, the role of circ-Foxo3 in esophageal squamous cell cancer (ESCC) is not clear. In this study, circ-Foxo3 was lowly expressed in cell lines and ESCC tissues. Meanwhile, overexpression of circ-Foxo3 inhibited cell growth, migration, and invasion, whether in vivo or in vitro. Mechanically, we found a potential miRNA target, miR-23a, which negatively correlated with circ-Foxo3 in ESCC. Then, a luciferase assay confirmed the relationship between the circ-Foxo3 and miRNA. Moreover, circ-Foxo3 upregulation of PTEN occurred through "sponging" miR-23a. Taken together, these results indicated that the circ-Foxo3/miR-23a/PTEN pathway was critical for inhibiting the ESCC progression. This may provide a promising target for treat ESCC.

Cost-effectiveness of Different Intra-articular Drug Injections on Wrist and/or Elbow Compromise in Patients With Established Rheumatoid Arthritis: Short-term Results of an Open Observational Evaluation.

BACKGROUND: Intra-articular (IA) injections are an integral part of the management of rheumatoid arthritis (RA). However, there are few reports regarding the association between drug effectiveness and cost. OBJECTIVES: The aim of this study was to assess the cost-effectiveness of various combinations of IA injections of betamethasone, hyaluronic acid (HA) or etanercept for oligoarthritis in RA. METHODS: Seventy RA patients were assigned to 4 groups according to the IA injection drug(s): betamethasone alone, betamethasone + etanercept, betamethasone + HA, or etanercept alone. Data for the following were collected before and after IA injection: erythrocyte sedimentation rate, C-reactive protein (CRP), disease activity score in 28 joints calculated with CRP, and patient global visual analog scale. In addition, power Doppler ultrasonography and gray-scale ultrasonography scores were obtained for synovitis, and passive range of motion of joints was measured. RESULTS: Sixty-eight RA patients completed the trial. Compared with patients given etanercept alone, the visual analog scale, power Doppler ultrasonography, and gray-scale ultrasonography scores of each of the other groups were significantly better at each time point. At 1 month, the passive range of motion of joints in patients given betamethasone + HA was significantly better than that of each of the other groups. Synovial hyperplasia improved significantly in all groups, but less so in those given etanercept alone. All other clinical parameters of the 4 groups were similar. The costs per joint for the betamethasone-alone, betamethasone + etanercept, betamethasone + HA, and etanercept-alone groups were, respectively, $7.55, $181.77, $42.68, and $174.22. CONCLUSIONS: Intra-articular injection of betamethasone alone was the most cost-effective treatment for oligoarthritis of RA. Betamethasone combined with HA injection resulted in the best improvement in joint function.

[The role of human lysozyme-like protein 4 in fertilization and its enzymatic properties].

OBJECTIVE: To elucidate the possible role of human lysozyme-like protein 4 (LYZL4) in fertilization and characterize its enzymatic properties. METHODS: The localization of LYZL4 in human spermatozoa was investigated by immunofluorescence staining, the sources of LYZL4 on the sperm surface examined by RT-PCR, and the role of LYZL4 in fertilization assessed by the zona-free hamster egg penetration test. The recombinant plasmid pPIC9K-LYZL4 was constructed and its expression induced with methanol after transformed into competent Pichia pastoris GS115. The recombinant LYZL4 protein (rLYZL4) was purified from the fermentation supernatant and subsequently identified by Western blot. The hyaluronan binding ability of rLYZL4 was determined by ELISA and the muramidase activity, hyaluronidase activity, and free radical scavenging ability examined by spectrophotometric methods. RESULTS: Immunodetection with a specific antiserum localized LYZL4 on the acrosomal membrane of mature spermatozoa, which was exclusively secreted from the testis and epididymis as shown by RT-PCR. Immunoneutralization of LYZL4 significantly decreased the number of human spermatozoa bound to zona-free hamster eggs in a dose-dependent manner in vitro. The recombinant protein was expressed successfully by the P. pastoris strain GS115. Purified rLYZL4 exhibited a potent hyaluronan binding ability and a strong free radical scavenging ability but no muramidase or hyaluronidase activity. CONCLUSIONS: LYZL4 secreted from the testis and epididymis is localized on the acrosomal membrane of mature spermatozoa and plays a role in sperm-egg binding as well as in binding hyaluronan and scavenging free radicals, which suggests that it might be a multi-functional molecule contributive to sperm protection and sperm-egg binding.

The establishment and evaluation of a swine model of deinagkistrodon acutus snakebite envenomation.

OBJECTIVE: To establish a preclinical large-animal model of Deinagkistrodon acutus snakebite envenomation and evaluate its feasibility. METHODS: The venom of D. acutus (0 mg/kg, 1 mg/kg, 2 mg/kg, 5 mg/kg, or 10 mg/kg) was injected into the left biceps femoris of 11 male pigs. Then, the circumferences of the limbs were regularly measured, and changes in muscle injury biomarkers, blood parameters, coagulation function, vital organ function and injury biomarkers were regularly detected. At 24 h after venom injection, the animals were euthanized, and the pathological damage to the vital organs mentioned above was evaluated. RESULTS: The two pigs receiving 10 mg/kg and 5 mg/kg snake venom died at 8 h and 12 h after injection, respectively. The remaining pigs were equally divided into 0 mg/kg, 1 mg/kg, and 2 mg/kg snake venom groups, and all of them survived to 24 h after injection. Compared with the pigs receiving 0 mg/kg snake venom, the pigs receiving 1 mg/kg or 2 mg/kg snake venom exhibited significant abnormities, including limb swelling; increased muscle injury biomarker creatine kinase (CK) and coagulation function indicators prothrombin time and D-dimer; and decreased blood routine indicator platelet and coagulation function indicator fibrinogen. Moreover, significant abnormalities in myocardial and cerebral function and injury biomarkers in the heart, brain, liver, kidney and intestine were also observed. In particular, the abnormalities mentioned above were significantly obvious in those pigs receiving 2 mg/kg snake venom. Pathological evaluation revealed that the morphology of muscle, heart, brain, liver, kidney, and intestine in those pigs receiving 0 mg/kg snake venom was normal; however, pathological damage was observed in those pigs receiving 1 mg/kg and 2 mg/kg snake venom. Similarly, the pathological damage was more severe in those pigs receiving 2 mg/kg snake venom. CONCLUSION: The intramuscular injection of 2 mg/kg D. acutus venom seems to be an optimal dose for examining the preclinical efficacy of existing and novel therapeutics for treating D. acutus envenomation in pigs.

Effects of aquatic versus land-based exercise on irisin and fibroblast growth factor 21 expression and triiodothyronine and free fatty acid levels in elderly women.

BACKGROUND: This study investigated the impacts of exercise on irisin and fibroblast growth factor 21 (FGF-21) expression, as well as triiodothyronine (T3 ) and free fatty acid (FFA) levels in elderly women. METHODS: Thirty women aged 65 to 70 years (10 per group) were randomly assigned to aquatic exercise, land exercise, and control groups. The aquatic and land groups engaged in 3 exercise sessions per week (60 min/session) for 16 weeks. The intensity was progressively increased every 4 weeks. RESULTS: Irisin and FGF-21 levels significantly increased in the aquatic exercise group. In the posttest, the aquatic exercise group had the highest irisin levels. Significant findings were observed for irisin and FGF-21 for the main effect between aquatic and band exercise groups (p<0.05 for both), the main effect between measurement times (p<0.01 and p<0.001, respectively), and the interaction effect (p<0.05 and p<0.001, respectively). The irisin level was significantly higher in the aquatic than in the land group 30 minutes after the last session (p<0.05). In both exercise groups, T3 levels were significantly higher 30 minutes after the final session (p<0.05) than before the program. The FFA level was significantly higher in the aquatic exercise group than the others. In the aquatic group, FFA levels were significantly higher 30 minutes after both the first (p<0.01) and the last (p<0.001) session compared to pre-program values. CONCLUSION: Differences in exercise type and environment can promote fat metabolism by stimulating hormonal changes that induce brown fat activity and browning.

[Role of computed tomographic angiography in the evaluations of dural arteriovenous fistulas with pulsatile tinnitus as initial symptom].

OBJECTIVE: To evaluate the sensitivity and specificity of computed tomographic angiography ( CTA) for dural arteriovenous fistulas ( DAVFs) in patients presenting with pulsatile tinnitus( PT). METHODS: The clinical and imaging data were collected for all patients undergoing CTA for PT from 2008 to 2012. Nine PT patients with DAVFs confirmed by digital subtraction angiography ( DSA) and 9 age and gender-matched control PT patients without DAVFs were selected. The CTA images were blindly analyzed by two experienced neuroradiologists for the following signs: asymmetric venous collaterals in extracranial space , asymmetric attenuation of internal jugular vein ( IJV) , asymmetric external carotid artery( ECA) branches, "shaggy" appearance of dural venous sinus, multiple transcalvarial channels, enlarged foramen spinosum, asymmetric cavernous sinus and enlarged cortical veins. RESULTS: The sensitivities of the following DAVFs signs were quite different: asymmetric attenuation of IJV ( 89% ) , asymmetric venous collaterals ( 89%) , asymmetric ECA branches ( 78%) , shaggy dural venous sinus ( 67%) , multiple transcalvarial channels (67%), enlarged foramen spinosum (56%), stenosis of venous sinus (33%) and asymmetric cavernous sinus ( 2 2 % ) . The presence of asymmetric attenuation of IJV , asymmetric ECA branches, shaggy dural venous sinus, multiple transcalvarial channels and asymmetric cavernous sinus all demonstrated a highly specificity of 100% while the presence of asymmetric venous collaterals and enlarged foramen spinosum had a specificity of 89% . The presence of stenosis of venous sinus revealed a specificity of 78%. Enlarged cortical veins were all absent. CONCLUSION: CTA may be used as a screening examination for DA VFs in PT patients. The presence of asymmetric venous collaterals, asymmetric attenuation of UV,asymmetric ECA branches, shaggy dural venous sinus and multiple transcalvarial channels has a high sensitivity and specificity for diagnosis. Enlarged ECA branches usually serve as DA VFs feeders.Meanwhile, DA VF should be considered in PT patients when multiple transcalvarial channels and enlarged foramen spinosum are detected on high-resolution CT of temporal bone.

Clinical value of SLC12A9 for diagnosis and prognosis in colorectal cancer.

OBJECTIVE: The goal of the study is to assess the clinical value and the potential mechanism of SLC12A9 combing transcriptome and single cell sequencing data. METHODS: In this study, the expression level and the receiver operating characteristic curve analysis of SLC12A9 in CRC and normal tissue were analyzed in multiple data cohort. The standardized mean difference (SMD) calculation and the summary receiver operating characteristic (SROC) analysis were performed further to detect its diagnostic ability and expression level. KM survival analysis was performed to assess the prognosis value of SLC12A9. The expression level of SLC12A9 in different clinical characteristics was analyzed to explore the clinical value. Single cell data was studied to reveal the potential mechanism of SLC12A9. The correlation analysis of immunoinfiltration was performed to detect the potential immune cell related to SLC12A9. The nomogram was drawn to assess the probable mortality rate of CRC patient. RESULTS: We found that SLC12A9 was significantly up-regulated with the moderate diagnostic value in CRC. Patients with overexpressed SLC12A9 had a worse prognosis. SLC12A9 was related to Age, Pathologic N stage, Pathologic M stage, Lymphatic invasion and Pathologic stage (p < 0.05). The 1, 3 and 5-year survival rates of patient named TCGA-G4-6309 are 0.959, 0.897 and 0.827. PCR also showed that SLC12A9 was overexpressed in CRC comparing with normal tissue. CONCLUSION: In conclusion, our study comprehensively analyzed the clinical value of SLC12A9 and its potential mechanism, as well as immune cell infiltration, which may accelerate the diagnosis and improve the prognosis of CRC.

A Practical Deep Learning Model in Differentiating Pneumonia-Type Lung Carcinoma from Pneumonia on CT Images: ResNet Added with Attention Mechanism.

OBJECTIVE: We aim to develop a deep neural network model to differentiate pneumonia-type lung carcinoma from pneumonia based on chest CT scanning and evaluate its performance. MATERIALS AND METHODS: We retrospectively analyzed 131 patients diagnosed with pneumonia-type lung carcinoma and 171 patients with pneumonia treated in Beijing Hospital from October 2019 to February 2021. The average age was 68 (±15) years old, and the proportion of men (162/302) was slightly more than that of women (140/302). In this study, a deep learning based model UNet was applied to extract lesion areas from chest CT images. Lesion areas were extracted and classified by a designed spatial attention mechanism network. The model AUC and diagnostic accuracy were analyzed based on the results of the model. We analyzed the accuracy rate, sensitivity, and specificity and compared the results of the model to the junior and senior radiologists and radiologists based on the model. RESULTS: The model has a good efficiency in detecting pneumonia-like lesions (6.31 seconds/case). The model accuracy rate, sensitivity, and specificity were 74.20%, 60.37%, and 89.36%, respectively. The junior radiologist's accuracy rate, sensitivity, and specificity were 61.00%, 48.08%, and 75.00%, respectively. The senior radiologist's accuracy rate, sensitivity, and specificity were 65.00%, 51.92%, and 79.17%, respectively. The results of junior radiologists based on the model were improved (76.00% for accuracy rate, 62.75% for sensitivity, and 89.80% for specificity). The results of senior radiologists based on the model were also improved (78.00% for accuracy rate, 64.71% for sensitivity, and 91.84% for specificity) and the diagnostic accuracy of which was statistically higher than other groups (P < 0.05). Based on the lesion texture diversity and the lesion boundary ambiguity, the algorithm produced false-positive samples (13.51%). CONCLUSION: This deep learning model could detect pneumonia-type lung carcinoma and differentiate it from pneumonia accurately and efficiently.

Diagnostic Value of Artificial Intelligence Based on CT Image in Benign and Malignant Pulmonary Nodules.

Objective: To evaluate the diagnostic value of artificial intelligence-assisted CT imaging in benign and malignant pulmonary nodules. Methods: The CT scan screening of pulmonary nodules from November 2018 to November 2020 was retrospectively collected. The diagnosis of pulmonary nodules and surgical treatment were performed. A total of 194 nodules in 152 patients with clear pathological results were observed. All patients underwent CT examination to analyze the consistency of the results of artificial intelligence, physician reading according to imaging features, multidisciplinary team work (MDT) diagnosis, and postoperative pathological results; the diagnostic efficacy of different diagnostic methods for solitary pulmonary nodules and the differences of ROC curve and AUC were analyzed. The accuracy, specificity, sensitivity, positive predictive value, negative predictive value, false negative rate, and false positive rate of different diagnostic methods for pulmonary nodules were calculated, and the ROC curves of different diagnostic methods were plotted. Results: The accuracy, sensitivity, specificity, and Youden index of artificial intelligence (AI) were 89.69%, 92.98%, 65.22%, and 58.20%; the accuracy, sensitivity, specificity, and Youden index of physician reading were 85.57%, 88.30%, 65.22%, and 53.52%; the accuracy, sensitivity, specificity, and Youden index of MDT were 96.91%, 98.25%, 86.96%, and 85.21%, respectively. The kappa values of artificial intelligence, physician reading, and MDT were 0.541, 0.437, and 0.852, and the AUC was 0.768, 0.791, and 0.926, respectively (P < 0.001). The average detection time of pulmonary nodules in the AI group, manual reading group, and MAT group was (145 ± 97) s, (534 ± 297) s, and (421 ± 128) s (P < 0.001). Conclusion: Artificial intelligence pulmonary nodule detection system can improve the coincidence rate and accuracy of early diagnosis of lung cancer, shorten the average detection time, and provide more accurate information for clinical decision-making.

Application Value of Spectral CT Imaging in Quantitative Analysis of Early Lung Adenocarcinoma.

Objective: To investigate the clinical value of gemstone energy spectral CT imaging for the quantitative analysis of early lung adenocarcinoma. Methods: 76 cases of pulmonary ground-glass nodules pathologically confirmed as precancerous lesion and early lung cancer (including pure ground-glass nodules in 46 cases and mixed ground-glass nodules in 30 cases) underwent chest CT scan first and then underwent contrast-enhanced gemstone energy spectral CT to get arterial phase images, venous phase images, and delayed phase images. All the lesions were set the region of interest (ROI). Cases of the pure ground-glass nodule (pGGN) were measured at the maximum level of lesions, cases of the mixed ground-glass nodule (mGGN) were measured in two areas of ground-glass and solid components, CT value and iodine concentrations of lesions in three-phase scanning were separately measured, and at the same time, iodine concentrations of the thoracic aorta were also measured. The normalized iodine concentrations (NICs) were calculated, that is, the ratio of iodine concentrations of lesions and the thoracic aorta. CT values of lesions were also measured at each stage of 70 keV. All the quantitative data were expressed by the mean ± standard deviation, and paired t-test was used for pairwise comparison. Results: In 76 cases, in the spectral CT imaging mode, the NIC value of solid components of the GGN was 0.33 ± 0.16 in the arterial phase (AP), 0.52 ± 0.25 in the venous phase (VP), and 0.58 ± 0.34 in the delayed phase (DP). There were significant differences of P values of NICs between each two phases in both solid component cases and the ground-glass component cases in AP/VP, VP/DP, and AP/DP (P < 0.05); there were no statistically significant P values of CT values between each two phases in three-period enhanced CT in both the solid component cases and the ground-glass component cases in AP/VP, VP/DP, and AP/DP (P > 0.05). Conclusion: Gemstone energy spectral CT with quantitative imaging can dynamically reflect the enhancement features of the pulmonary GGN.

Integrated Analysis of Genomic and Transcriptomic Profiles Identified the Role of GTP Binding Protein-4 (GTPBP4) in Breast Cancer.

Purpose: To explore the significance of GTP-binding protein 4 (GTPBP4) in breast cancer. Methods: Firstly, GTPBP4 expression analysis was performed in TIMER and UALCAN databases. Subsequently, the TCGA cohort and multiple Gene Expression Omnibus Cohorts were used as validation for GTPBP4 expression. Besides, we also evaluated the diagnostic value of GTPBP4 in TCGA Cohort and multiple GEO Cohorts. The predictive effect of GTPBP4 in breast cancer was then assessed using survival analysis. Then we look at the role of GTPBP4 in the immune milieu and create a Nomogram to help patients with breast cancer understand their prognosis. Finally, in vitro tests were carried out to look at GTPBP4 expression and function in breast cancer cell lines. Results: GTPBP4 is an independent breast cancer prognostic factor that is upregulated in the disease (p < 0.05). Enrichment analysis showed that GTPBP4 was associated with multiple functions and pathways. In addition, GTPBP4 is associated with a variety of immune cell types (p < 0.05). PCR assay showed that GTPBP4 expression was up-regulated in breast cancer cell lines. The activity, migration, and proliferation of breast cancer cells were considerably reduced after GTPBP4 knockdown in the CCK-8, Transwell, and Scratch assays. Conclusions: Our research discovered a new breast cancer biomarker that can be used as a guide for breast cancer diagnosis and treatment.

Micropatterns and peptide gradient on the inner surface of a guidance conduit synergistically promotes nerve regeneration in vivo.

Both of the surface topographical features and distribution of biochemical cues can influence the cell-substrate interactions and thereby tissue regeneration in vivo. However, they have not been combined simultaneously onto a biodegradable scaffold to demonstrate the synergistic role so far. In this study, a proof-of-concept study is performed to prepare micropatterns and peptide gradient on the inner wall of a poly (D,L-lactide-co-caprolactone) (PLCL) guidance conduit and its advantages in regeneration of peripheral nerve in vivo. After linear ridges/grooves of 20/40 µm in width are created on the PLCL film, its surface is aminolyzed in a kinetically controlled manner to obtain the continuous gradient of amino groups, which are then transferred to CQAASIKVAV peptide density gradient via covalent coupling of glutaraldehyde. The Schwann cells are better aligned along with the stripes, and show a faster migration rate toward the region of higher peptide density. Implantation of the nerve guidance conduit made of the PLCL film having both the micropatterns and peptide gradient can significantly accelerate the regeneration of sciatic nerve in terms of rate, function recovery and microstructures, and reduction of fibrosis in muscle tissues. Moreover, this nerve conduit can also benefit the M2 polarization of macrophages and promote vascularization in vivo.

Combining Single-Cell and Transcriptomic Data Revealed the Prognostic Significance of Glycolysis in Pancreatic Cancer.

Background: Pancreatic cancer (PC), the most common fatal solid malignancy, has a very dismal prognosis. Clinical computerized tomography (CT) and pathological TNM staging are no longer sufficient for determining a patient's prognosis. Although numerous studies have suggested that glycolysis is important in the onset and progression of cancer, there are few publications on its impact on PC. Methods: To begin, the single-sample gene set enrichment analysis (ssGSEA) approach was used to quantify the glycolysis pathway enrichment fraction in PC patients and establish its prognostic significance. The genes most related to the glycolytic pathway were then identified using weighted gene co-expression network analysis (WGCNA). The glycolysis-associated prognostic signature in PC patients was then constructed using univariate Cox regression and lasso regression methods, which were validated in numerous external validation cohorts. Furthermore, we investigated the activation of the glycolysis pathway in PC cell subtypes at the single-cell level, performed a quasi-time series analysis on the activated cell subtypes and then detected gene changes in the signature during cell development. Finally, we constructed a decision tree and a nomogram that could divide the patients into different risk subtypes, according to the signature score and their different clinical characteristics and assessed the prognosis of PC patients. Results: Glycolysis plays a risky role in PC patients. Our glycolysis-related signature could effectively discriminate the high-risk and low-risk patients in both the trained cohort and the independent externally validated cohort. The survival analysis and multivariate Cox analysis indicated this gene signature to be an independent prognostic factor in PC. The prognostic ROC curve analysis suggested a high accuracy of this gene signature in predicting the patient prognosis in PC. The single-cell analysis suggested that the glycolytic pathway may be more activated in epithelial cells and that the genes in the signature were also mainly expressed in epithelial cells. The decision tree analysis could effectively identify patients in different risk subgroups, and the nomograms clearly show the prognostic assessment of PC patients. Conclusion: Our study developed a glycolysis-related signature, which contributes to the risk subtype assessment of patients with PC and to the individualized management of patients in the clinical setting.

Selective Adhesion and Directional Migration of Endothelial Cells Guided by Cys-Ala-Gly Peptide Density Gradient on Antifouling Polymer Brushes.

Selective adhesion and directional migration of endothelial cells (ECs) on biomaterials is critical to realize the rapid endothelialization. In this study, a Cys-Ala-Gly (CAG) peptide density gradient is generated on homogeneous cell-resisting poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) brushes by immersing the brushes in a complementary gradient solution of CAG and competitive mercapto-terminated methoxyl poly(ethylene glycol). The adhesion and spreading of smooth muscle cells (SMCs) is impaired effectively on the gradient surface. About six folds of adherent ECs over SMCs are achieved at the position (10 mm) of highest CAG density on the gradient surface in a co-culture condition. Due to the gradient cues, ECs migrate fastest with the best directionality of 86.7% at the middle of the gradient, leading to the maximum net displacement as well.

Temperature-Gating Titania Nanotubes Regulate Migration of Endothelial Cells.

External stimuli-responsive biomaterials represent a type of promising candidates for addressing the complexity of biological systems. In this study, a platform based on the combination of temperature-sensitive polymers and a nanotube array was developed for loading sphingosine 1-phosphate (S1P) and regulating the migration of endothelial cells (ECs) at desired conditions. The localized release dosage of effectors could be controlled by the change of environmental temperature. At a culture temperature above the lower critical solution temperature, the polymer "gatekeeper" with a collapsed conformation allowed the release of S1P, which in turn enhanced the migration of ECs. The migration rate of single cells was significantly enhanced up to 58.5%, and the collective migration distance was also promoted to 25.1% at 24 h and 33.2% at 48 h. The cell morphology, focal adhesion, organization of cytoskeleton, and expression of genes and proteins related to migration were studied to unveil the intrinsic mechanisms. The cell mobility was regulated by the released S1P, which would bind with the S1PR1 receptor on the cell membrane and trigger the Rho GTPase pathway.

Biodegradable Anisotropic Microparticles for Stepwise Cell Adhesion and Preparation of Janus Cell Microparticles.

The biomimetic anisotropic particles have different physicochemical properties on the opposite two sides, enabling diverse applications in emulsion, photonic display, and diagnosis. However, the traditional anisotropic particles have a very small size, ranging from submicrons to a few microns. The design and fabrication of anisotropic macron-sized particles with new structures and properties is still challenging. In this study, anisotropic polycaprolactone (PCL) microparticles well separated with each other were prepared by crystallization from the dilute PCL solution in a porous 3D gelatin template. They had fuzzy and smooth surfaces on each side, and a size as large as 70 µm. The fuzzy surface of the particle adsorbed significantly larger amount of proteins, and was more cell-attractive regardless of the cell types. The particles showed stronger affinity toward fibroblasts over hepatocytes, which paved a new way for cell isolation merely based on the surface morphology. After a successive seeding process, Janus cell microparticles with fibroblasts and endothelial cells (ECs) on each side were designed and obtained by making use of the anisotropic surface morphology, which showed significant difference in EC functions in terms of prostacyclin (PGl2) secretion, demonstrating the unique and appealing functions of this type of anisotropic microspheres.

Inflammatory activation of human serum albumin- or ovalbumin-modified chitosan particles to macrophages and their immune response in human whole blood.

Nanomaterials have been extensively used in the biomedical field. These nanoscale objects may either promote or restrain immune responses depending on their surface characteristics and compositions. In this study, chitosan (CS) particles prepared using an emulsion-crosslinking method were modified with different amounts of human serum albumin (HSA) and ovalbumin (OVA), resulting in four types of modified CS particles, i.e. CS@HSA-10, CS@HSA-57, CS@OVA-13 and CS@OVA-65, respectively. They had a similar size of about 150 nm in a dry state, and were swollen 2-3 fold in PBS. No significant cytotoxicity was determined toward in vitro cultured RAW264.7 and THP-1 cells. However, all the modified CS particles, in particular the OVA-modified ones (CS@OVA-13 and CS@OVA-65), induced significantly higher secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) compared with the negative control. In human whole blood, CS@OVA-13 and CS@OVA-65 were phagocytosed with a significantly higher ratio by granulocytes and monocytes, leading to the higher secretion of TNF-α, IL-1ß and IL-8, and a larger extent of platelet activation than CS@HSA-10 and CS@HSA-57, respectively.

[Analysis of the relationship between living habit and cervical instability in adolescent patients with neck pain].

OBJECTIVE: To investigate the relationship between living habit and cervical instability in adolescent patients with neck pain. METHODS: Fifty-nine adolescent patients with neck pain(neck pain group) and seventeen healthy teenagers (control group) were recruited and divided into two groups, and clinical information, living habit were collected. In addition, all people were taken lateral, hyperextension and hyperflexion radiography to analyze relationship between living habit and cervical instability. RESULTS: There was no obvious difference in age, height, weight and body mass index between two groups. The neck pain group using cellphone time per day is longer than control group, while control group had more exercise time than neck pain group(P<0.01). The incidence of instability in neck pain group was significantly higher than that in control group(P<0.01). In hyperflexion, angular displacement(AD) of neck pain group in vertebral body between C3-C4, C4-C5 and C5-C6 was significantly higher than that of control group. In neck pain group, AD of hyperflexion was higher than that of hyperextension on C4-C5(P<0.01), and AD of hyperextension is higher than that of hyperflexion on C6-C7(P<0.05). In neck pain group, AD of hyperextension on C4-C5 was positively correlated with time of using cellphone every day(r=0.275, P=0.035). And AD of hyperflexion was significantly positive correlated with time of using cellphone(r=0.577, P<0.001), but was negatively correlated with exercise time(r=-0.279, P=0.032). The AD of hyperflexion on C5-C6 was negatively correlated with exercise time every day(r=-0.292, P=0.025), AD of hyperextension was negatively correlated with time of using computer every day(r=-0.262, P=0.045). CONCLUSIONS: Adolescent neck pain patients had more time to use cellphone than normal teens every day, and exercise time is less than healthy teenagers, and occurrence rate of cervical instability is higher on C3-C4, C4-C5, C5-C6 segment. The longer daily exercise time, the smaller C4-C5 and C5-C6 AD values; the longer cellphone usage every day, the greater C4-C5 AD values.

Regulating the migration of smooth muscle cells by a vertically distributed poly(2-hydroxyethyl methacrylate) gradient on polymer brushes covalently immobilized with RGD peptides.

The gradient localization of biological cues is of paramount importance to guide directional migration of cells. In this study, poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate)-block- poly(2-hydroxyethyl methacrylate) (P(HEMA-co-GMA)-b-PHEMA) brushes with a uniform underneath P(HEMA-co-GMA) layer and a gradient thickness of PHEMA blocks were prepared by using surface-initiated atom-transfer radical polymerization and a dynamically controlled polymerization process. The polymer chains were subsequently functionalized with the cell-adhesive arginine-glycine-aspartic acid (RGD) peptides by reaction with the glycidyl groups, and their structures and properties were characterized by X-ray photoelectron spectrometry (XPS), quartz crystal microbalance with dissipation (QCM-D) and air contact angle. Adhesion and migration processes of smooth muscle cells (SMCs) were then studied. Compared with those on the sufficiently exposed RGD surface, the cell adhesion and mobility were well maintained when the RGD peptides were localized at 18.9 nm depth, whereas the adhesion, spreading and migration rate of SMCs were significantly impaired when the RGD peptides were localized at a depth of 38.4 nm. On the RGD depth gradient surface, the SMCs exhibited preferential orientation and enhanced directional migration toward the direction of reduced thickness of the second PHEMA brushes. Half of the cells were oriented within ±â€¯30° to the x-axis direction, and 72% of the cells moved directionally at the optimal conditions. Cell adhesion strength, arrangement of cytoskeleton, and gene and protein expression levels of adhesion-related proteins were studied to corroborate the mechanisms, demonstrating that the cell mobility is regulated by the complex and synergetic intracellular signals resulted from the difference in surface properties. STATEMENT OF SIGNIFICANCE: Cell migration is of paramount importance for the processes of tissue repair and regeneration. So far, the gradient localization of biological cues perpendicular to the substrate, which is the usual case for the biological signaling molecules to locate in ECM in vivo, has been scarcely studied, and has not been used to guide the directional migration of cells. In this study, we prepare a depth gradient of RGD peptides along the polymer chains, which is used to guide the directional migration of SMCs after a second hydrophilic bock is prepared in a gradient manner. For the first time the directional migration of SMCs is achieved under the guidance of a depth gradient of RGD ligands. The mechanisms of different cell migration abilities are further discussed based on the results of cell adhesion, cell adhesion force, cytoskeleton alignment and expression of relative proteins and genes. This work paves a new strategy by fabricating a gradient polymer brushes with immobilized bioactive molecules to dominate the directional cell migration, and elucidates the mechanisms underlining the biased migration along RGD depth localization gradients, shedding a light for the design of novel biomaterials to control and guide cell migration and invasion.