Clinimetric Properties of the "FIND-NEEDS" to Screen Geriatric Conditions.

INTRODUCTION: Comprehensive geriatric assessment (CGA) is used to thoroughly assess and identify complex healthcare problems among older adults. However, administration of CGA is time-consuming and labor intensive. A simple screening tool with the mnemonic "FIND-NEEDS" was developed to quickly identify common geriatric conditions. The present study was to evaluate the clinimetric properties of the FIND-NEEDS. METHODS: The participants comprised first-visiting older adults aged 65 years and above (and who were able to communicate by themselves or with the help of a caregiver) who were assessed (October to December, 2021) using the FIND-NEEDS and CGA at geriatric outpatient clinics of a tertiary, referred medical center. The FIND-NEEDS was examined for its criterion-related validity and compared with the CGA results. Two types of scoring (summed score and binary score) of FIND-NEEDS and CGA were analyzed using Spearman correlation, sensitivity and specificity, and area under receiver operating characteristic curve (AUC). RESULTS: The mean age of the 114 outpatients was 78.3 ± 7.6 years, and 79 (69.3%) were female. The internal consistency was excellent when using all FIND-NEEDS items, and was acceptable when using domain scores. Exploratory factor analysis showed that most of the FIND-NEEDS domain scores had factor loadings higher than 0.3. Intercorrelations of binary scores between domains of FIND-NEEDS and CGA showed most domains were moderately correlated. The overall correlation of summed scores between FIND-NEEDS and CGA was high. The FIND-NEEDS summed score was moderately correlated with CGA score (r = 0.494; p < 0.001), and the binary score showed excellent correlation (r = 0.944; p < 0.001). When using the CGA score as the gold standard, the FIND-NEEDS showed excellent AUC (0.950), sensitivity (1.00), and specificity (0.90). DISCUSSION/CONCLUSION: The present study demonstrated that the FIND-NEEDS had acceptable clinimetric properties to screen for geriatric problems among older adults. Further in-depth assessment and care plan can then be conducted afterwards.

Active secretion of IL-33 from astrocytes is dependent on TMED10 and promotes central nervous system homeostasis.

Interleukin-33 (IL-33), secreted by astrocytes, regulates the synapse development in the spinal cord and hippocampus and suppresses autoimmune disease in the central nervous system (CNS). However, the mechanism of unconventional protein secretion of this cytokine remains unclear. In this study, we found that IFN-γ promotes the active secretion of IL-33 from astrocytes, and the active secretion of IL-33 from cytoplasm to extracellular space was dependent on interaction with transmembrane emp24 domain 10 (TMED10) via the IL-1 like cytokine domain in astrocytes. Knockout of Il-33 or its receptor St2 induced hippocampal astrocyte activation and depressive-like disorder in naive mice, as well as increased spinal cord astrocyte activation and polarization to a neurotoxic reactive subtype and aggravated passive experimental autoimmune encephalomyelitis (EAE). Our results have identified that IL-33 is actively secreted by astrocytes through the unconventional protein secretion pathway facilitated by TMED10 channels. This process helps maintain CNS homeostasis by inhibiting astrocyte activation.

Cellular Mechanisms of Psoriasis Pathogenesis: A Systemic Review.

Psoriasis is a common inflammatory skin disease characterized by abnormal proliferation of epidermal keratinocytes and massive infiltration of inflammatory cells. Many kinds of cells, including keratinocytes, T lymphocytes, dendritic cells, neutrophils, and macrophages, are reported to play critical roles in the pathogenesis and progression of psoriasis. However, to date, the role of each kind of cell in the pathogenesis and development of psoriasis has not been systematically reviewed. In addition, although antibodies developed targeting cytokines (e.g. IL-23, IL-17A, and TNF-α) released by these cells have shown promising results in the treatment of psoriasis patients, these targeted antibodies still do not cure psoriasis and only provide short-term relief of symptoms. Furthermore, long-term use of these antibodies has been reported to have adverse physical and psychological effects on psoriasis patients. Therefore, gaining a deeper understanding of the cellular and molecular pathogenesis of psoriasis and providing new thoughts on the development of psoriasis therapeutic drugs is of great necessity. In this review, we summarize the roles of various cells involved in psoriasis, aiming to provide new insights into the pathogenesis and development of psoriasis at the cellular level and hoping to provide new ideas for exploring new and effective psoriasis treatments.

Topical application of TOPK inhibitor OTS514 suppresses psoriatic progression by inducing keratinocytes cell cycle arrest and apoptosis.

T-LAK cell-oriented protein kinase (TOPK) potently promotes malignant proliferation of tumour cells and is considered as a maker of tumour progression. Psoriasis is a common inflammatory skin disease characterized by abnormal proliferation of keratinocytes. However, the role of TOPK in psoriasis has not been well elucidated. This study aims to investigate the expression and role of TOPK in psoriasis, and the role of TOPK inhibitor in psoriasis attenuation. Gene Expression Omnibus datasets derived from psoriasis patients and psoriatic model mice were screened for analysis. Skin specimens from psoriasis patients were collected for TOPK immunohistochemical staining to investigate the expression and localization of TOPK. Next, psoriatic mice model was established to further confirm TOPK expression pattern. Then, TOPK inhibitor was applied to investigate the role of TOPK in psoriasis progression. Finally, cell proliferation assay, apoptosis assay and cell cycle analysis were performed to investigate the potential mechanism involved. Our study showed that TOPK was upregulated in the lesions of both psoriasis patients and psoriatic model mice, and TOPK levels were positively associated with psoriasis progression. TOPK was upregulated in psoriatic lesions and expressed predominantly by epidermal keratinocytes. In addition, TOPK levels in epidermal keratinocytes were positively correlated with epidermal hyperplasia. Furthermore, topical application of TOPK inhibitor OTS514 obviously alleviated disease severity and epidermal hyperplasia. Mechanismly, inhibiting TOPK induces G2/M phase arrest and apoptosis of keratinocytes, thereby attenuating epidermal hyperplasia and disease progression. Collectively, this study identifies that upregulation of TOPK in keratinocytes promotes psoriatic progression, and inhibiting TOPK attenuates epidermal hyperplasia and psoriatic progression.

IL-33 and HMGB1 modulate the progression of EAE via oppositely regulating each other.

Interleukin-33 (IL-33) and high mobility group box 1 (HMGB1) have been reported to play crucial and distinct roles in experimental autoimmune encephalomyelitis (EAE). However, little is known about their interaction in the progression of EAE. In this study, the dynamic expression and release of IL-33 and HMGB1 in different stages of EAE in vivo, and their interaction in vitro were explored. We found that HMGB1 was dominant in pre-onset stage of EAE, while IL-33 was dominant in peak stage. Moreover, both blockade of extracellular HMGB1 in the central nervous system (CNS) and conditional knockout of HMGB1 in astrocytes decreased IL-33 release. HMGB1 promoted the release of IL-33, while IL-33 reduced the release of HMGB1 from primary astrocytes in vitro. Taken together, IL-33 and HMGB1 in the CNS jointly participate in the EAE progression and the inhibitory effect of IL-33 on HMGB1 may be involved in the self-limiting of EAE.

IL-33 Contributes to the Pathological Changes of Hair Follicles in Psoriasis: A Potential Target for Psoriatic Alopecia.

Purpose: IL-33 is constitutively expressed in skin tissues. Alopecia, a T cells-driven disorder of the hair follicles (HFs), is a common complication in the development of psoriasis. However, the role of IL-33 in psoriatic alopecia remains uncovered. Here, we investigated the roles of IL-33 in inducing pathological changes of hair follicles in psoriasis. Patients and Methods: Clinical samples and imiquimod (IMQ)-induced psoriatic mice samples were used to investigate the pathological changes and T-cell infiltration of HFs. By using immunohistochemistry staining, the distribution and expression alteration of IL-33 in HFs were determined. Next, by using IL-33 and ST2 knockout mice, we investigated the role of IL-33/ST2 axis in the pathological changes of HFs in psoriasis. Meanwhile, recombinant IL-33 protein was subcutaneous injected to confirm its effect. Finally, RNA sequencing was used to clarify the genes and signaling pathways that involved in this process. Differentially expressed genes were further verified by RT-PCR in cultured HFs in vitro. Results: We found that the pathological changes of HFs and T cells infiltration in imiquimod-induced psoriatic mice were similar to that in psoriasis patients. The IL-33 positive keratinocytes in the outer root sheath of HFs were increased in both psoriasis patients and psoriatic model mice compared with the controls. By using gene knockout mice, we found that the pathological changes and T cell infiltration were attenuated in IL-33-/- and ST2-/- psoriatic model mice. In addition, subcutaneous injection of recombinant IL-33 exacerbated the pathological changes of HFs and T cell infiltration. RNA sequencing and RT-RCR revealed that IL-33 upregulated the transcription of genes related to keratinocytes proliferation and T lymphocytes chemotaxis. Conclusion: Our study identifies that IL-33 promotes the pathological changes of HFs in psoriasis, which contributes to psoriatic alopecia. Inhibition of IL-33 may be a potential therapeutic approach for psoriatic alopecia.

HMGB1 from Astrocytes Promotes EAE by Influencing the Immune Cell Infiltration-Associated Functions of BMECs in Mice.

High mobility group box 1 (HMGB1) has been reported to play an important role in experimental autoimmune encephalomyelitis (EAE). Astrocytes are important components of neurovascular units and tightly appose the endothelial cells of microvessels by their perivascular endfeet and directly regulate the functions of the blood-brain barrier. Astrocytes express more HMGB1 during EAE while the exact roles of astrocytic HMGB1 in EAE have not been well elucidated. Here, using conditional-knockout mice, we found that astrocytic HMGB1 depletion decreased morbidity, delayed the onset time, and reduced the disease score and demyelination of EAE. Meanwhile, there were fewer immune cells, especially pathogenic T cells infiltration in the central nervous system of astrocytic HMGB1 conditional-knockout EAE mice, accompanied by up-regulated expression of the tight-junction protein Claudin5 and down-regulated expression of the cell adhesion molecules ICAM1 and VCAM1 in vivo. In vitro, HMGB1 released from astrocytes decreased Claudin5 while increased ICAM1 and VCAM1 expressed by brain microvascular endothelial cells (BMECs) through TLR4 or RAGE. Taken together, our results demonstrate that HMGB1 derived from astrocytes aggravates EAE by directly influencing the immune cell infiltration-associated functions of BMECs.

Allograft or Recipient ST2 Deficiency Oppositely Affected Cardiac Allograft Vasculopathy via Differentially Altering Immune Cells Infiltration.

The role of IL-33/ST2 signaling in cardiac allograft vasculopathy (CAV) is not fully addressed. Here, we investigated the role of IL-33/ST2 signaling in allograft or recipient in CAV respectively using MHC-mismatch murine chronic cardiac allograft rejection model. We found that recipients ST2 deficiency significantly exacerbated allograft vascular occlusion and fibrosis, accompanied by increased F4/80+ macrophages and CD3+ T cells infiltration in allografts. In contrast, allografts ST2 deficiency resulted in decreased infiltration of F4/80+ macrophages, CD3+ T cells and CD20+ B cells and thus alleviated vascular occlusion and fibrosis of allografts. These findings indicated that allografts or recipients ST2 deficiency oppositely affected cardiac allograft vasculopathy/fibrosis via differentially altering immune cells infiltration, which suggest that interrupting IL-33/ST2 signaling locally or systematically after heart transplantation leads different outcome.

Identification of Tumor Tissue of Origin with RNA-Seq Data and Using Gradient Boosting Strategy.

BACKGROUND: Cancer of unknown primary (CUP) is a type of malignant tumor, which is histologically diagnosed as a metastatic carcinoma while the tissue-of-origin cannot be identified. CUP accounts for roughly 5% of all cancers. Traditional treatment for CUP is primarily broad-spectrum chemotherapy; however, the prognosis is relatively poor. Thus, it is of clinical importance to accurately infer the tissue-of-origin of CUP. METHODS: We developed a gradient boosting framework to trace tissue-of-origin of 20 types of solid tumors. Specifically, we downloaded the expression profiles of 20,501 genes for 7713 samples from The Cancer Genome Atlas (TCGA), which were used as the training data set. The RNA-seq data of 79 tumor samples from 6 cancer types with known origins were also downloaded from the Gene Expression Omnibus (GEO) for an independent data set. RESULTS: 400 genes were selected to train a gradient boosting model for identification of the primary site of the tumor. The overall 10-fold cross-validation accuracy of our method was 96.1% across 20 types of cancer, while the accuracy for the independent data set reached 83.5%. CONCLUSION: Our gradient boosting framework was proven to be accurate in identifying tumor tissue-of-origin on both training data and independent testing data, which might be of practical usage.

A machine learning framework to trace tumor tissue-of-origin of 13 types of cancer based on DNA somatic mutation.

Carcinoma of unknown primary (CUP), defined as metastatic cancers with unknown cancer origin, occurs in 3-5 per 100 cancer patients in the United States. Heterogeneity and metastasis of cancer brings great difficulties to the follow-up diagnosis and treatment for CUP. To find the tissue-of-origin (TOO) of the CUP, multiple methods have been raised. However, the accuracies for computed tomography (CT) and positron emission tomography (PET) to identify TOO were 20%-27% and 24%-40% respectively, which were not enough for determining targeted therapies. In this study, we provide a machine learning framework to trace tumor tissue origin by using gene length-normalized somatic mutation sequencing data. Somatic mutation data was downloaded from the Data Portal (Release 28) of the International Cancer Genome Consortium (ICGC), and 4909 samples for 13 cancers was used to identify primary site of cancers. Optimal results were obtained based on a 600-gene set by using the random forest algorithm with 10-fold cross-validation, and the average accuracy and F1-score were 0.8822 and 0.8886 respectively across 13 types of cancer. In conclusion, we provide an effective computational framework to infer cancer tissue-of-origin by combining DNA sequencing and machine learning techniques, which is promising in assisting clinical diagnosis of cancers.

A Deep Learning Framework to Predict Tumor Tissue-of-Origin Based on Copy Number Alteration.

Cancer of unknown primary site (CUPS) is a type of metastatic tumor for which the sites of tumor origin cannot be determined. Precise diagnosis of the tissue origin for metastatic CUPS is crucial for developing treatment schemes to improve patient prognosis. Recently, there have been many studies using various cancer biomarkers to predict the tissue-of-origin (TOO) of CUPS. However, only a very few of them use copy number alteration (CNA) to trance TOO. In this paper, a two-step computational framework called CNA_origin is introduced to predict the tissue-of-origin of a tumor from its gene CNA levels. CNA_origin set up an intellectual deep-learning network mainly composed of an autoencoder and a convolution neural network (CNN). Based on real datasets released from the public database, CNA_origin had an overall accuracy of 83.81% on 10-fold cross-validation and 79% on independent datasets for predicting tumor origin, which improved the accuracy by 7.75 and 9.72% compared with the method published in a previous paper. Our results suggested that the autoencoder model can extract key characteristics of CNA and that the CNN classifier model developed in this study can predict the origin of tumors robustly and effectively. CNA_origin was written in Python and can be downloaded from https://github.com/YingLianghnu/CNA_origin.

A novel drug repurposing approach for non-small cell lung cancer using deep learning.

Drug repurposing is an attractive and pragmatic way offering reduced risks and development time in the complicated process of drug discovery. In the past, drug repurposing has been largely accidental and serendipitous. The most successful examples so far have not involved a systematic approach. Nowadays, remarkable advances in drugs, diseases and bioinformatic knowledge are offering great opportunities for designing novel drug repurposing approach through comprehensive understanding of drug information. In this study, we introduced a novel drug repurposing approach based on transcriptomic data and chemical structures using deep learning. One strong candidate for repurposing has been identified. Pimozide is an anti-dyskinesia agent that is used for the suppression of motor and phonic tics in patients with Tourette's Disorder. However, our pipeline proposed it as a strong candidate for treating non-small cell lung cancer. The cytotoxicity of pimozide against A549 cell lines has been validated.

A comparison study of SARS-CoV-2 IgG antibody between male and female COVID-19 patients: A possible reason underlying different outcome between sex.

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in China at the end of 2019 has spread throughout the world and caused many thousands of deaths. The previous study reported a higher severe status rate and mortality rate in male patients in China. However, the reason underlying this difference has not been reported. The convalescent plasma containing a high level of SARS-CoV-2 immunoglobulin G (IgG) antibody has been used in clinical therapy and achieved good effects in China. In this study, to compare the differences of the SARS-CoV-2 IgG antibody between male and female patients, a total number of 331 patients confirmed SARS-CoV-2 infection were enrolled. The serum of these patients was collected during hospitalization and detected for the SARS-CoV-2 IgG antibody. Our data showed that the concentration of IgG antibody in mild, general, and recovering patients showed no difference between male and female patients. In severe status, compared with male patients, there were more female patients having a relatively high concentration of serum SARS-CoV-2 IgG antibody. In addition, the generation of IgG antibody in female patients was stronger than male patients in disease early phase. Our study identified a discrepancy in the SARS-CoV-2 IgG antibody level in male and female patients, which may be a potential cause leading to a different outcome of Coronavirus Disease 2019 between sex.

Technology-embedded health education on nutrition for middle-aged and older adults living in the community.

Mobile technology provides young adults important support for self-directed learning, but whether there is related support for older adults is not clear. This study aims to determine whether 1) nutrition education combined with mobile technology-supported teaching improves knowledge of and self-efficacy for a healthy diet; 2) if adults who reported reviewing the electronic course material or searching health information online, showed significantly greater progress in knowledge of and self-efficacy for a healthy diet than did those who did not adopt the electronic support. A total of 35 middle-aged and older adults were recruited from the community. Enrollees who were unable to read, who participated in the course fewer than five times, who did not take the post-test, or who did not return complete questionnaires at the pre-test were excluded. Overall, 21 participants were finally analyzed, and 14 participated in the qualitative investigation. The study interventions included three traditional nutrition lectures and three touch-screen tablet computer lessons to access the Internet and nutrition applications. Structured and semi-structured questionnaires were used to collect both quantitative and qualitative data and record participants' Internet use conditions at home. Participants' nutrition knowledge significantly improved (meanpost-pre = 1.19, p = 0.001) and their self-efficacy about a healthy diet showed marginal improvement (meanpost-pre = 0.22, p = 0.07). Nutrition knowledge was positively correlated with their intensity of surfing the Internet (r = 0.46, p < 0.05), or reviewing the electronic course material (r = 0.48, p < 0.05) but not correlated with reviewing paper course material (r = 0.19, p = 0.09). Qualitative results showed that participants reported feeling freshness, joyfulness, and great achievement because of the combined course material. Technology-supported learning combined with traditional health education might provide great opportunities for positive behavioral change, even in older adults without any previous Internet experience.

The Role of Leisure Activities in the Relationship Between Marital Transition in Later Midlife and Psychological Well-Being Trajectories.

This study examined the levels and rates of changes in psychological well-being for middle-aged adults of different statuses or marital transitions. The moderating effects of different leisure activities were also tested. Longitudinal data on 1,270 persons aged 50 to 65 years at baseline from the Taiwan longitudinal study on aging were analyzed. Adults who were stably unmarried or unpartnered reported worse mental health at baseline, but their psychological well-being improved over time. The trajectory of depressive symptoms fluctuated markedly in adults who became widowed during our observation period. Engagement in physical, cognitive, or social activities was significantly associated with participants' psychological well-being. Participation in religious activities was significantly associated with life satisfaction and decreased depressive symptoms for those undergoing bereavement. Findings from this study suggest that social and physical activities, among the four selected leisure activities, have the greatest association between decreasing depressive symptoms and increasing life satisfaction, respectively. Religious activities, in particular, may improve psychological well-being in bereaved middle-aged and older adults.

Suicide Reporting on Front Pages of Major Newspapers in Taiwan Violating Reporting Recommendations Between 2001 and 2012.

Media reporting can lead to the acceptance of incorrect ideas and information by the general public, which in turn can impact behavior. A number of studies have found that suicide reporting in the media can lead to an increase in the suicide rate or copycat suicides. Therefore, the aims of this study are to (a) investigate front-page reporting of suicide in four major newspapers in Taiwan; (b) investigate violation of recommendations for reporting suicide by the four major newspapers in Taiwan; and (c) investigate the impact of important events and government policy on front-page reporting of suicide (i.e., the effect of the Apple Daily newspaper entering the market, and the start of media monitoring). We carried out content analysis of reports of suicide on the front pages of the four newspapers with the highest rates of readership in Taiwan (China Times, Liberty Times, Apple Daily, and United Daily News) between 2001 and 2012. We used the chi-squared test, chi-squared test for trend, and analysis of variance to analyze trends in violation of reporting recommendations and potential associated factors. We found that (a) suicide was most commonly reported as a front-page headline and as a whole-page spread by the four major newspapers in Taiwan, with many reports including photographs; (b) reporting of suicide by the four major newspapers in Taiwan frequently violated World Health Organization (WHO) recommendations for suicide reporting in the media; (c) following the entry of the Apple Daily newspaper into the Taiwanese market, reporting approaches to suicide by the other three major newspapers changed to more sensational photos and texts; and (d) monitoring of suicide reporting by the Taiwan Suicide Prevention Center appears to have been only partially effective. In summary, reporting of suicide by the four major newspapers in Taiwan could be further improved. Effective regulation of suicide reporting by newspapers could reduce the impact of this reporting on readers. In addition, regular assessment of observance of the WHO recommendations for suicide reporting by newspapers is an important part of suicide prevention.

Effects of parenting role and parent-child interaction on infant motor development in Taiwan Birth Cohort Study.

BACKGROUND: Previous studies have rarely focused on healthy infants' motor development, and nationwide birth cohort studies in Taiwan are limited. It has been shown that parent-child interactions significantly influence infant motor development and the effect of mother-infant attachment on infant development is stronger than father-infant attachment. However, it is not well understood that whether the mother-infant or father-infant interaction has the confounding effect on infant motor development. AIMS: To understand healthy infant motor development in Taiwan; and to investigate the effects of parenting roles and parent-child interactions on infant motor development. METHODS: Data were derived from the 1st through the 2nd waves of the Taiwan Birth Cohort Study-Pilot Database. Infants were classified into two categories (complete or incomplete development) according to their developmental milestones. Generalized estimating equations (GEE) and random effects models were used to clarify the possible long-term effects. RESULTS: The rate of infants who completed development in 6 months was 30.50%; however the rate was increased in 18 month-old children (80.01%). A mother's perceived infant care competence was the most important factor for infant motor development. "Whether or not the infant was the only baby in the family" and "parent-child interaction" had slightly significant effect on infant motor development. CONCLUSIONS: In conclusion, the mother's perceived competence must be strengthened and parent-infant interactions should be emphasized on a daily basis.

Influence of stressors and possible pathways of onset of seventh graders' suicidal ideation in urban and rural areas in Taiwan.

BACKGROUND: Suicide is the second leading cause of death among young people in Taiwan. However, few studies have investigated children's suicidal ideation, and longitudinal studies are particularly rare. The purposes of this study were: (1) to describe the proportion of students with suicidal ideation in one month and incidence of suicidal ideation in the 7th graders (the first year of junior high school) living in urban and rural areas; (2) to realize the influence of perceived stressors on the onset of 7th graders' suicidal ideation; and (3) to explore possible pathways through which trigger factors and perceived stressors lead to suicidal ideation. METHODS: A total of 1,589 students were followed from grade 4 to grade 7. Logistic regression was then used to investigate the influence of perceived stressors on the onset of suicidal ideation in grade 7. Structural equation modeling was used to analyze possible pathways through which trigger factors led to increased pressure from certain stressors which in turn resulted in suicidal ideation. RESULTS: The proportion of students with suicidal ideation in one month was 2 to 3 times higher in both areas compared to that in elementary school. However, the incidence in the rural area showed a large increase from 10.2% and 9.5% in grades 5 and 6 to 15.5% in grade 7. Urban-rural difference was observed. Important stressors and pathways of suicidal ideation differ between urban and rural areas. CONCLUSIONS: This study showed that the influential stressors in urban and rural areas might be different. Thus, interventions focused on coping skills for regional specific stressors and trigger factors could be beneficial in the transition time.

[In vitro studies on the growth and proliferation characteristics of rat bone mesenchymal stem cells].

OBJECTIVE: This study was to investigate the growth and proliferation characteristics of rat bone mesenchymal stem cells (BMSCs) isolated by the method of whole bone marrow culture and to explore the effect of cell inoculation density and incubation period on cell proliferation, with an aim to provide multipotential seed cells for preventing from degenerative disease. METHODS: Bone mesenchymal stem cells were isolated by the method of whole bone marrow culture and then cultured in vitro. The cell morphologic features were observed by inverted microscope. The cell surface antigens were identified by flow cytometry. The effect of cell inoculation density and culture period on cell growth and proliferation was explored by analyzing the characteristics of a ten-day cell growth curve in 96-well plates. RESULTS: Flow cytometry results showed the detection rates for CD29, CD34 and CD45 were 97.68% (7607/7788), 7.93% (661/8340) and 2.76% (215/7788) respectively, which was consistent with the expression characteristics of BMSCs surface antigens. BMSCs became uniform after three cell passages, existing in a typical shape of whirlpool or radial colony. The senescent cells started to appear at 7(th) passage, and more senescent cells were found at 10(th) passage. The growth curve for moderate inoculation density was typically S-shaped. Lag phase was found during the first two days, and logarithm growth phase was in the following three days. Plateau phase started from the 6(th) day and cell numbers decreased slightly from the 8(th) day. CONCLUSION: The whole bone marrow culture is an effective way to obtain BMSCs. A moderate inoculation density was more advantageous to cell proliferation, by which more seed cells could be obtained.

[Effects of drynaria total flavonoid on osteogenic differentiation of bone marrow mesenchymal stem cells at different glucose concentrations: experiment with rats].

OBJECTIVE: To study the effects of drynaria total flavonoid on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) at different glucose concentrations. METHODS: BMSCs of SD rats were isolated, cultivated in vitro, and divided into 6 groups to be induced to differentiate into osteoblasts under different conditions: (1) low glucose control group, (2) high glucose control group, (3) low glucose classical induction group (sodium glycerophosphate+vitamin C+dexamethasone), (4) high glucose classical induction group (sodium glycerophosphate+vitamin C+dexamethasone), (5) low glucose+drynaria total flavonoid group, and (6) high glucose with drynaria total flavonoid group. Alkaline phosphate (ALP) test kit was used to examine the level of ALP. The ALP staining positive rate was examined with modified calcium cobalt method. Alizarin red staining was adopted to observe the number of calcium nodes. Immunohistochemistry was used to detect type I collagen level. Advanced glycosylation end products (AGEs) were tested by ELISA. RESULTS: The A value indicating the ALP activity, ALP staining positive rate, calcium node number, and type I collagen expression score of the low glucose+drynaria total flavonoid group were (0.439±0.024), 48.7%, (9.75±1.71) nodes/HP, and (2.21±0.07) respectively, all significantly higher than those of the sodium glycerophosphate+vitamin C+dexamethasone [(0.385±0.029), 35.0%, (6.25±0.96) nodes/HP, and (1.93±0.13) respectively, all P<0.05]. The A value, ALP staining positive rate, calcium node number, and type I collagen expression score of the high glucose with drynaria total flavonoid group were (0.352±0.022), 25.3%, (4.50±1.29)/HP, and (1.70±0.03) respectively, all significantly higher than those of the sodium glycerophosphate+vitamin C+dexamethasone [(0.139±0.013), 22.7%, (3.25±1.50)/HP, and (1.28±0.27) respectively, all P<0.05]. The AGE expression levels of the high glucose classical induction group and high glucose+drynaria total flavonoid group were both significantly higher than those of the low glucose classical induction group and low glucose+drynaria total flavonoid group (both P<0.05). There were no significant differences in the AGE level among the low glucose control, low glucose classical induction, and low glucose+drynaria total flavonoid groups (all P<0.05); and among the high glucose control, high glucose classical induction, and high glucose+drynaria total flavonoid groups (all P<0.05). However, the AGE levels of the high glucose groups were all significantly higher than those of the corresponding low glucose groups (all P<0.05). Glucose increased the AGE levels dose- and time-dependently. The concentrations of AGEs were significantly negatively correlated with the expression of type I collagen (r=-0.410, P<0.05). CONCLUSIONS: Drynaria total flavonoid promotes the osteogenic differentiation of BMSCs and relieves the inhibitory effect of osteogenic differentiation by glucose at high concentration. Thus drynaria total flavonoid may provide a potential therapy for diabetic osteoporosis.