Effect of Hyaluronic Acid on Cytokines and Immune Cells Change in Patients of Knee Osteoarthritis

Purpose To evaluate the changes of cytokines and immune cells after Intra-articular hyaluronic acid（IAHA）injections in patients with knee osteoarthritis (KOA).Patients and Methods Sixteen patients were included in the study, with a total of 63 IAHA injections. The Numeric Rating Scale (NRS) and Lysholm scores were evaluated at each visit. The immune cells and 14 cytokines of synovial fluid were analyzed at each visit. The association between immune cells and cytokines were examined Results IL-6 and IL-8 were the most common cytokines in the synovial fluid of KOA patients. The synovial fluid was orchestrated by macrophages (69%) and Lymphocytes (18%). Neutrophils were less to count of the total cell population (<2%). The cytokines decreased significantly after the first injection and then tended to be stable. Lymphocytes increased a lot, while Macrophages decreased in the early stage, then increased after multiple injections. The proposition of M1 decreased in the early stage, then increased after multiple injections, while M2 increased consistently. M1 and M2 were positively associated with IL-6 and IL-8.Conclusion The synovial fluid of KOA patients was orchestrated by macrophages (69%) and Lymphocytes (18%) and cytokines like IL-6 and IL-8. IAHA may play an anti-inflammatory functional role through the decreased production of IL-6 and IL-8 by macrophages through polarization. The results from this study provided new evidence for IAHA treatment in KOA patients, and therapies targeting pathogenic cytokines and immune cells might be used to attenuate the knee joint inflammation and release pain.Trial registrationChiCTR2100050133; date registered 17 August 2021.

Differentiation of Tumorigenic C6 Glioma Cells Induced by Enhanced IL-6 Signaling

Background and objectives: Cancer stem cells (CSCs) are obstacles to cancer therapy due to their therapeutic resistance, ability to initiate neoplasia, and roles in tumor relapse and metastasis. Efforts have been made to cure CSCs, such as the use of differentiation therapy, which induces cancer stem-like cells to undergo differentiation and decrease their tumorigenicity. Interleukin 6 (IL-6) upregulates the expression of glial fibrillary acidic protein (GFAP) in C6 glioma cells, indicating that it is able to induce the differentiation of these cells. The C6 glioma cell line forms a high percentage of cancer stem-like cells, leading us to speculate whether IL-6 signaling could modulate the differentiation of tumorigenic C6 glioma cells. However, we observed that IL-6 alone could not efficiently induce the differentiation of these cells. Therefore, different IL-6 signaling elicitors, including IL-6 alone, a combination of IL-6 and soluble IL-6 receptor (IL-6/sIL-6R), and tumor necrosis factor-α (TNF-α) plus IL-6/sIL-6R (TNF-α/IL-6/sIL-6R), were evaluated for their potential use in differentiation therapy. Materials and Methods: The potential of IL-6 signaling elicitors in differentiation therapy were examined by assessing changes in biomarker levels, the rate of cell proliferation, and tumorigenicity, respectively. Results: Enhanced IL-6 signaling could effectively induce C6 glioma cell differentiation, as determined by observed variations in the expression of differentiation, cell cycle, and stem cell biomarkers. Additionally, the total cell population and the tumorigenicity of glioma cells were all considerably reduced after TNF-α/IL-6/sIL-6R treatment. Conclusions: Our findings provide evidence that enhanced IL-6 signaling can efficiently promote tumorigenic C6 glioma cells to undergo differentiation.

Mathematical models incorporating a multi-stage cell cycle replicate normally-hidden inherent synchronization in cell proliferation

We present a suite of experimental data showing that cell proliferation assays, prepared using standard methods thought to produce asynchronous cell populations, persistently exhibit inherent synchronization. Our experiments use fluorescent cell cycle indicators to reveal the normally hidden cell synchronization, by highlighting oscillatory subpopulations within the total cell population. These oscillatory subpopulations would never be observed without these cell cycle indicators. On the other hand, our experimental data show that the total cell population appears to grow exponentially, as in an asynchronous population. We reconcile these seemingly inconsistent observations by employing a multi-stage mathematical model of cell proliferation that can replicate the oscillatory subpopulations. Our study has important implications for understanding and improving experimental reproducibility. In particular, inherent synchronization may affect the experimental reproducibility of studies aiming to investigate cell cycle-dependent mechanisms, including changes in migration and drug response.

Mathematical models incorporating a multi-stage cell cycle replicate normally-hidden inherent synchronisation in cell proliferation

We present a suite of experimental data showing that cell proliferation assays, prepared using standard methods thought to produce asynchronous cell populations, persistently exhibit inherent synchronisation. Our experiments use fluorescent cell cycle indicators to reveal the normally-hidden cell synchronisation by highlighting oscillatory subpopulations within the total cell population. These oscillatory subpopulations would never be observed without these cell cycle indicators. On the other hand, our experimental data show that the total cell population appears to grow exponentially, as in an asynchronous population. We reconcile these seemingly inconsistent observations by employing a multi-stage mathematical model of cell proliferation that can replicate the oscillatory subpopulations. Our study has important implications for understanding and improving experimental reproducibility. In particular, inherent synchronisation may affect the experimental reproducibility of studies aiming to investigate cell cycle-dependent mechanisms, including changes in migration and drug response.

Population regulation in microbial consortia using dual feedback control

An ongoing area of study in synthetic biology has been the design and construction of synthetic circuits that maintain homeostasis at the population level. Here, we are interested in designing a synthetic control circuit that regulates the total cell population and the relative ratio between cell strains in a culture containing two different cell strains. We have developed a dual feedback control strategy that uses two separate control loops to achieve the two functions respectively. By combining both of these control loops, we have created a population regulation circuit where both the total population size and relative cell type ratio can be set by reference signals. The dynamics of the regulation circuit show robustness and adaptation to perturbations in cell growth rate and changes in cell numbers. The control architecture is general and could apply to any organism for which synthetic biology tools for quorum sensing, comparison between outputs, and growth control are available.

Inhibition of SENP5 by Cucurbitacin B Suppresses Cell Growth and Promotes Apoptosis in Osteosarcoma Cells

Purpose: To investigate the effect of cucurbitacin B on the expression of SUMO-specific proteases (SENP5).Methods: Effect of cucurbitacin B (10-50 mg/mL) on viability of U2OS and Saos-2 cells was determined in a plate reader by recording absorbance at 570 nm. In Western blot analysis, bicinconinic acid (BCA) method was used to determine protein concentration. Flow cytometry was employed to measure DNA content.Results: Ccucurbitacin B treatment inhibits the expression of SENP5 in U2OS and Saos-2 osteosarcoma cells in a dose- and time-dependent manner. Significant inhibition (p = 0.005) of SENP5 expression was observed at 50 mg/ml from day 10, reaching a maximum on day 20. It also induced a significant decrease (p = 0.005) in mRNA and protein levels of SENP5. The decrease in mRNA and protein levels of SENP5 led to decrease in proliferation of U2OS and Saos-2 cells. The 48 h cell cultures containing 50 mg of cucurbitacin B caused induction of apoptosis in 54.72 ± 5.42 % of the total cell population in U2OS cells. Similarly, in Saos-2 cells, exposure to 50 mg/mL cucurbitacin B increased apoptotic rate from 9.86 ± 8.89 % for 10 mg/mL to 48.54 ± 14.5 % with 50 mg/mL of cucurbitacin B.Conclusion: Cucurbitacin B is a potential therapeutic strategy for the treatment of aggressive malignancy in osteosarcoma.Keywords: Cucurbitacin B, Osteosarcoma, SUMO-specific proteases, Cell proliferation, Apoptosis, Malignancy

TOTAL CELL NUMBER IN FETAL BRAIN

In this study the material comprises brains from three aborted fetuses and two fullterm infants who died at birth.The gestational ages ranged from the 22nd week to term. All cases were without malformations, known chromosomal abnormality, hydrops, and systemic infections, and all had normal birth weights with fetal growth indices (observed birth weight/expected mean birth weight) between 0.9 - 1.05. The preliminary results show a five fold increase in the total cell population in the marginal zone/cortical plate, MZ/CP (future neocortex), from week 22 until term. In the transient subplate zone, SP, the total cell number was more than doubled from week 22 to week 30-35, and then decreased towards term. In the intermediate zone, IZ (future white matter), the total cell population was doubled from week 22 until term. The total cell number in the entricular/subventricular zone, VZ/SZ (germinal matrix), was reduced by a factor of five from week 22 until term. A histological differentiation between neurons and glial cells was not possible. The optical fractionator was used to estimate the total cell population in four characteristic developmental zones in the human fetal brain. Fetal brain tissue undergoes considerable and rather unpredictable shrinkage during fixation. However, using the fractionator principle it is possible to eliminate this problem, provided that the structure of interest (one brain hemisphere) is fully intact.

Neurochemical phenotype and birthdating of specific cell populations in the chick retina

The chick embryo is one of the most traditional models in developing neuroscience and its visual system has been one of the most exhaustively studied. The retina has been used as a model for studying the development of the nervous system. Here, we describe the morphological features that characterize each stage of the retina development and studies of the neurogenesis period of some specific neurochemical subpopulations of retinal cells by using a combination of immunohistochemistry and autoradiography of tritiated-thymidine. It could be concluded that the proliferation period of dopaminergic, GABAergic, cholinoceptive and GABAceptive cells does not follow a common rule of the neurogenesis. In addition, some specific neurochemical cell groups can have a restrict proliferation period when compared to the total cell population.

SNP-A Karyotyping Demonstrates a High Incidence of Segmental Uniparental Disomy in Patients with CMML and Shows Impact of Newly Identified Chromosomal Aberrations on Clinical Course.

The WHO classification recognizes chronic myelomonocytic leukemia (CMML) as an overlap syndrome sharing clinical and histomorphologic features with both MDS and MPD. Unlike in CML or classical MPD, which are often characterized by recurrent translocations or activating mutations, only 1/3 of CMML patients harbor lesions (e.g. balanced translocations that include PDGFb as a fusion partner). Also frequent are unbalanced aberrations similar to those seen in classical MDS. Of note is that IPSS assigns prognosis only to a portion of patients with CMML (those with &gt;10% of blasts and/or high WBC counts). Since chromosomal defects have a major impact on the diagnosis and prognosis of myeloid malignancies, it is likely that cytogenetic methods with higher resolution and an ability to detect uniparental disomy (UPD) could explain clinical heterogeneity and point to potential therapeutic targets in CMML. We applied 250K SNP-arrays (SNP-A) to examine karyotype and identify previously cryptic defects in patients with low grade and advanced forms of CMML. SNP-A allows for detection of clones spanning 25–50% of total cell population, and fidelity of LOH calls is &gt;99% as shown by analysis of chromosome (chr.) X in males. Any deletions, duplications, and/or UPD found by SNP-A in 76 controls or those on internet databases were considered copy number variants (CNV) and excluded from analysis. In total, we studied 77 patients with CMML; 42/77 showed abnormal MC, including most often lesions commonly associated with MDS/CMML, such as +8 (N=5) and +19 (N=2). DNA was available for SNP-A karyotyping in 46 patients. Abnormal karyotype was detected in 19/46 patients (40%) by MC compared to 37/46 patients (79%) by SNP-A. Examples of newly detected lesions included microdeletions of chrs. 12 and 7, and various micro-duplications/deletions. Remarkably, we found (perhaps in analogy to UPD9p seen in MPD) a high prevalence of segmental UPD, occurring in 20/47 patients (43%) with significant recurrence on chrs. 4 (N=4), 6 (N=4), and 11 (N=4). 7/20 patients had UPD as a sole or isolated abnormality. In 3 these patients, UPD11q was the sole contributing lesion while in one patient with UPD11, only one additional lesion, a small microdeletion of chr. 3, was found. Of note is that previously we have found also UPD11q in 4/29 patients with MDS/MPD-U. When we analyzed SNP-A results in CMML patients according to blast counts (CMML-1/2) and WBC (myeloproliferative type (MP) vs. myelodysplastic type (MD)), CMML-2 patients showed a higher frequency/more complex lesions, likely acquired in the process of transformation (1.5 vs. 2.2 avg. lesions). In addition to identifying abnormal overlapping/recurrent aberrations, SNP-A karyotyping has a potential clinical utility. When we stratified patients according to SNP-A detected lesions, we found a statistically significant difference between overall survival of patients with normal MC and normal SNP-A vs. those with normal MC but abnormal SNP-A (p=0.03, 40.2 vs. 7.3 months). In summary, SNP-A-based karyotyping complements MC and allows for precise definition of chr. aberrations in patients with CMML, including copy-neutral LOH. UPD is common in CMML and overlapping regions may point to potential causative genes.