KLF2 Orchestrates Pathological Progression of Infantile Hemangioma through Hemangioma Stem Cell Fate Decisions.

Infantile hemangioma (IH) is the most prevalent vascular tumor during infancy, characterized by a rapid proliferation phase of disorganized blood vessels and spontaneous involution. IH possibily arises from a special type of multipotent stem cells called hemangioma stem cells (HemSCs), which could differentiate into endothelial cells, pericytes, and adipocytes. However, the underlying mechanisms that regulate the cell fate determination of HemSCs remain elusive. Here, we unveil KLF2 as a candidate transcription factor involved in the control of HemSCs differentiation. KLF2 exhibits high expression in endothelial cells in proliferating IH but diminishes in adipocytes in involuting IH. Using a combination of in vitro culture of patient-derived HemSCs and HemSCs implantation mouse models, we show that KLF2 governs the proliferation, apoptosis and cell cycle progression of HemSCs. Importantly, KLF2 acts as a crucial determinant of HemSCs' fate, directing their differentiation toward endothelial cells while inhibiting adipogenesis. Knockdown of KLF2 induces a pro-adipogenic transcriptome in HemSCs, leading to impaired blood vessel formation and accelerated adipocyte differentiation. Collectively, our findings highlight KLF2 as a critical regulator controlling the progression and involution of IH by modulating HemSCs' cell fate decisions.

[Diagnosis and treatment of 11 patients with cevical spondylotic amyotrophy].

OBJECTIVE: To explore clinical features, treatment methods and clinical effects of cervical spondylosis with proximal muscular atrophy. METHODS: Eleven patients with proximal-type cervical spondylotic amyotrophy were retrospectively studied from September 2016 to November 2020, including 7 males and 4 females, aged 38 to 68 years old. Clinical symptoms, MRI and neuroelectrophysiological manifestations were analyzed, and patients were treated with conservative treatment or anterior cervical decompression fusion surgery, respectively. The efficacy was evaluated by manual muscle test (MMT) before and after treatment, and patients' satisfaction was followed up at the same time. RESULTS: All patients were followed up for 6 to 19 months. All 11 patients were unilateral, mainly manifested by atrophy of deltoid muscle, supraspinatus muscle and infraspinatus muscle, and may be accompanied by ipsilateral neck and shoulder pain at early stage. MRI showed lesions at C4,5, C5,6 segments were more common. Electrophysiological examination showed the affected muscle was denervated, and amplitude of compound muscle action potential (CMAP) of innervated nerve on the affected side was lower than that on the healthy side. All patients were obtained bone fusion. One patient who were underwent anterior cervical corpectomy and fusion (ACCF) occurred developed contralateral C5 nerve root paralysis after operation, which recovered completely after 10 weeks of symptomatic treatment. At 12 months after operation, the efficacy was evaluated according to MMT, 3 patients were treated conservatively, 2 patients excellent and 1 good;in 8 patients treated by operation, 3 patients were excellent, 4 good, and 1 moderate. CONCLUSION: The incidence of cervical spondylosis with proximal muscular atrophy is low, which is manifested as unilateral proximal muscle atrophy and may be accompanied by ipsilateral neck and shoulder pain in the early stage. Combined with MRI and neuroelectrophysiological examination, misdiagnosis could be reduced. In the early stage of disease, especially in the case of nucleus pulposus protrusion leading to nerve compression, conservative treatment could be taken. When the conservative treatment is ineffective or the pain cannot be tolerated, anterior decompression surgery is recommended, and the overall effect is satisfactory.

Investigation on the effect of ulinastatin on the apoptosis of vascular smooth muscle cells in rats with aortic dissection based on the Sirt1/FoxO3a pathway.

This study aimed to investigate the effects of ulinastatin on the apoptosis and (Sirt1/FoxO3a) pathway of vascular smooth muscle cells (VSMC) in aortic dissection (AD) rats. For this purpose a rat model of aortic dissection (AD) was constructed by giving drinking water containing 0.08% ß-aminopropionitrile (BAPN) to rats, HE staining was used to observe the pathological changes of the aorta in AD rats; the diseased blood vessels of AD rats were taken for primary culture and passage of VSMCs, the morphology of VSMCs was observed, and VSMCs were identify with immunofluorescence staining; VSMCs were treated with culture media containing 0, 1000, 2000, 3000, 4000, 5000, 6000, 7000 U/mL ulinastatin, and MTT kit was used to determine the effect of ulinastatin on VSMC proliferation in AD rats; the VSMC of AD rats were divided into blank group (normal culture), ulinastatin group (medium containing 5000 U/mL ulinastatin), Sirt1 inhibitor group (medium containing 1 µmol/L EX527), ulinastatin + Sirt1 inhibitor group (medium containing 5000 U/mL ulinastatin, 1 µmol/L EX527), flow cytometry was used to detect the VSMC apoptosis in each group, WB was used to detect the expression of VSMC apoptosis-related proteins and Sirt1/FoxO3a pathway-related proteins in each group. Findings suggested that the aortic wall of AD rats was thickened, and the dissection false cavity appeared; VSMC mostly presented different shapes such as triangles and stars, the immunofluorescence staining results showed that α-SMA was arranged in the cytoplasm in the form of myofilaments, showing green fluorescence, and the nucleus showed blue fluorescence, and the rate of positive cells was more than 95%; various doses of ulinastatin had a certain inhibitory effect on the proliferation of VSMC, and 5000 U/mL ulinastatin had a higher proliferation inhibition rate; compared with the blank group, the VSMC apoptosis rate, Caspase-3, Bax protein, Sirt1/FoxO3a pathway related protein expression in the ulinastatin group were significantly increased, and the Bcl-2 protein expression was significantly decreased (P<0.05), the VSMC apoptosis rate, Caspase-3, Bax protein, Sirt1/FoxO3a pathway related protein expression in the Sirt1 inhibitor group were significantly decreased, and the Bcl-2 protein expression was significantly increased (P<0.05); compared with the ulinastatin group, the VSMC apoptosis rate, Caspase-3, Bax protein, Sirt1/FoxO3a pathway related protein expression in the ulinastatin + Sirt1 inhibitor group were significantly decreased, and the Bcl-2 protein expression was significantly increased (P<0.05). It was concluded that ulinastatin can inhibit the proliferation of VSMCs in AD rats and promote their apoptosis, which may be achieved by activating the Sirt1/FoxO3a pathway.

PbrWRKY70 increases pear (Pyrus bretschneideri Rehd) black spot disease tolerance by negatively regulating ethylene synthesis via PbrERF1B-2.

Various pear plant cultivars exhibit diverse abilities to resist pear black spot disease (BSD), while the precise molecular mechanisms of resistance against pear BSD remain unclear. This study proposed a profound expression of a WRKY gene, namely PbrWRKY70, derived from Pyrus bretschneideri Rehd, within a BSD-resistant pear cultivar. Comparative analysis against the wild-type revealed that the overexpression of PbrWRKY70 engendered augmented BSD resistance of transgenic Arabidopsis thaliana and pear calli. Notably, the transgenic plants exhibited higher activities of superoxide dismutase and peroxidase, along with an elevated capacity to counteract superoxide anions via increased anti-O2-. Additionally, these plants displayed diminished lesion diameter, as well as reduced levels of hydrogen peroxide, malondialdehyde and 1-aminocyclopropane-1-carboxylic acid (ACC) contents. We subsequently demonstrated that PbrWRKY70 selectively bound to the promoter region of ethylene-responsive transcription factor 1B-2 (PbrERF1B-2), a potential negative regulator of ACC, thereby downregulating the expression of ACC synthase gene (PbrACS3). Consequently, we confirmed that PbrWRKY70 could enhance pear resistance against BSD by reducing ethylene production via modulation of the PbrERF1B-2-PbrACS3 pathway. This study established the pivotal relationship among PbrWRKY70, ethylene synthesis and pear BSD resistance, fostering the development of novel BSD-resistant cultivars. Furthermore, this breakthrough holds the potential to enhance pear fruit yield and optimize storage and processing during the later stages of fruit maturation.

A visual denaturation bubble-mediated strand exchange amplification and RGB visual analysis-based assay for quantitative detection of Helicobacter pylori in saliva.

Helicobacter pylori (H. pylori) is a class I carcinogen causing gastric cancer. Almost 50% of people on earth have been infected and it is worse in developing countries. Early diagnosis of H. pylori infection is the most important strategy for preventing the spread and worse consequences. H. pylori can be isolated from human saliva, and the sampling of saliva is easy and convenient. Therefore, we developed a visual denaturation bubble-mediated strand exchange amplification and RGB visual analysis-based assay for quantitative detection of H. pylori in saliva in this study. Under the optimized reaction temperature and time, the SEA reaction could be finished in 30 min with a simple reaction system and low dependency on equipment. The detection results could be qualitatively identified by the naked eye and quantitatively analyzed by a developed RGB visual analysis method. The limit of detection (LOD) of RGB visual analysis was 10.8 CFU/mL. This assay had good specificity and anti-interference capacity. In the artificial contamination test, the recovery rate of our assay was between 99.3% and 111.5%, with RSD values ranging from 1.7% to 3.5%. These indicated our assay also had good reliability in the detection of saliva. We believe this assay showed good potential for better non-invasive diagnosis of H. pylori infection.

A time-resolved fluorescent microsphere-lateral flow immunoassay strip assay with image visual analysis for quantitative detection of Helicobacter pylori in saliva.

Helicobacter pylori (H. pylori) is a kind of microaerobic and food-borne pathogen. More than 4.4 billion individuals have been infected by H. pylori and H. pylori was listed as Group I carcinogen by WHO in 1994. Considering the high infection rate of H. pylori and the limited medical resources, self-testing is helpful for diagnosis and timely treatment. Although the amount of H. pylori in human saliva is low, the sampling of saliva is simple and convenient compared with stomach, blood and stool samples. Therefore, H. pylori in human saliva can be an indicator for self-testing, and a sensitive and easy-to-use assay is necessary. In this study, we developed a time-resolved fluorescent microsphere-lateral flow immunoassay (TRFM-LFIA) strip assay with image visual analysis for detection of H. pylori in saliva. The detection of the TRFM-LFIA strip was easy to use and had a low dependency on equipment. With optimized preparation and detection parameters, the whole detection process could be finished in 8 min and the LOD by naked eyes was 102 CFU/mL. For quantitative analysis by image visual analysis, the LOD was as low as 1.05 CFU/mL in the linear range of 101-105 CFU/mL. Besides, the TRFM-LFIA strip also had good stability, reliability, repeatability and accuracy. All these advantages indicated that the TRFM-LFIA strips developed in this study had a good potential for self-testing for H. pylori infection.

Integration of 3D genome topology and local chromatin features uncovers enhancers underlying craniofacial-specific cartilage defects.

Aberrations in tissue-specific enhancers underlie many developmental defects. Disrupting a noncoding region distal from the human SOX9 gene causes the Pierre Robin sequence (PRS) characterized by the undersized lower jaw. Such a craniofacial-specific defect has been previously linked to enhancers transiently active in cranial neural crest cells (CNCCs). We demonstrate that the PRS region also strongly regulates Sox9 in CNCC-derived Meckel's cartilage (MC), but not in limb cartilages, even after decommissioning of CNCC enhancers. Such an MC-specific regulatory effect correlates with the MC-specific chromatin contacts between the PRS region and Sox9, highlighting the importance of lineage-dependent chromatin topology in instructing enhancer usage. By integrating the enhancer signatures and chromatin topology, we uncovered >10,000 enhancers that function differentially between MC and limb cartilages and demonstrated their association with human diseases. Our findings provide critical insights for understanding the choreography of gene regulation during development and interpreting the genetic basis of craniofacial pathologies.

Surface Phase Engineering Modulated Iron-Nickel Nitrides/Alloy Nanospheres with Tailored d-Band Center for Efficient Oxygen Evolution Reaction.

The oxygen evolution reaction (OER) plays a key role in many electrochemical energy conversion systems, but it is a kinetically sluggish reaction and requires a large overpotential to deliver appreciable current, especially for the non-noble metal electrocatalysts. In this study, the authors report a surface phase engineering strategy to improve the OER performance of transition metal nitrides (TMNs). The iron-nickel nitrides/alloy nanospheres (FeNi3 -N) wrapped in carbon are synthesized, and the optimized FeNi3 -N catalyst displays dual-phase nitrides on the surface induced by atom migration phenomenon, resulting from the different migration rates of metal atoms during the nitridation process. It shows excellent OER performance in alkaline media with an overpotential of 222 mV at 10 mA cm-2 , a small Tafel slope of 41.53 mV dec-1 , and long-term durability under high current density (>0.5 A cm-2 ) for at least 36 h. Density functional theory (DFT) calculations further reveal that the dual-phase nitrides are favorable to decrease the energy barrier, modulate the d-band center to balance the absorption and desorption of the intermediates, and thus promote the OER electrochemical performance. This strategy may shed light on designing OER and other catalysts based on surface phase engineering.

Preparation of Hollow Cobalt-Iron Phosphides Nanospheres by Controllable Atom Migration for Enhanced Water Oxidation and Splitting.

Transition metal phosphides (TMPs), especially the dual-metal TMPs, are highly active non-precious metal oxygen evolution reaction (OER) electrocatalysts. Herein, an interesting atom migration phenomenon induced by Kirkendall effect is reported for the preparation of cobalt-iron (Co-Fe) phosphides by the direct phosphorization of Co-Fe alloys. The compositions and distributions of the Co and Fe phosphides phases on the surfaces of the electrocatalysts can be readily controlled by Cox Fey alloys precursors and the phosphorization process with interesting atom migration phenomenon. The optimized Co7 Fe3 phosphides exhibit a low overpotential of 225 mV at 10 mA cm-2 in 1 m KOH alkaline media, with a small Tafel slope of 37.88 mV dec-1 and excellent durability. It only requires a voltage of 1.56 V to drive the current density of 10 mA cm-2 when used as both anode and cathode for overall water splitting. This work opens a new strategy to controllable preparation of dual-metal TMPs with designed phosphides active sites for enhanced OER and overall water splitting.

Overexpression of an apple LysM-containing protein gene, MdCERK1-2, confers improved resistance to the pathogenic fungus, Alternaria alternata, in Nicotiana benthamiana.

BACKGROUND: Lysin motif (LysM)-containing proteins are involved in the recognition of fungal and bacterial pathogens. However, few studies have reported on their roles in the defense responses of woody plants against pathogens. A previous study reported that the apple MdCERK1 gene was induced by chitin and Rhizoctonia solani, and its protein can bind to chitin. However, its effect on defense responses has not been investigated. RESULTS: In this study, a new apple CERK gene, designated as MdCERK1-2, was identified. It encodes a protein that shares high sequence identity with the previously reported MdCERK1 and AtCERK1. Its chitin binding ability and subcellular location are similar to MdCERK1 and AtCERK1, suggesting that MdCERK1-2 may play a role in apple immune defense responses as a pattern recognition receptor (PRR). MdCERK1-2 expression in apple was induced by 2 fungal pathogens, Botryosphaeria dothidea and Glomerella cingulate, but not by the bacterial pathogen, Erwinia amylovora, indicating that MdCERK1-2 is involved in apple anti-fungal defense responses. Further functional analysis by heterologous overexpression (OE) in Nicotiana benthamiana (Nb) demonstrated that MdCERK1-2 OE improved Nb resistance to the pathogenic fungus, Alternaria alternata. H2O2 accumulation and callose deposition increased after A. alternata infection in MdCERK1-2 OE plants compared to wild type (WT) and empty vector (EV)-transformed plants. The induced expression of NbPAL4 by A. alternata significantly (p < 0.01, n = 4) increased in MdCERK1-2 OE plants. Other tested genes, including NbNPR1, NbPR1a, NbERF1, and NbLOX1, did not exhibit significant changes after A. alternata infection in OE plants compared to EV or WT plants. OE plants also accumulated more polyphenols after A. alternata infection. CONCLUSIONS: Heterologous MdCERK1-2 OE affects multiple defense responses in Nb plants and increased their resistance to fungal pathogens. This result also suggests that MdCERK1-2 is involved in apple defense responses against pathogenic fungi.

Development process of traumatic heterotopic ossification of the temporomandibular joint in mice.

PURPOSE: The exact development process underlying traumatic heterotopic ossification of the temporomandibular joint (THO-TMJ) is largely unclear. In this study, we try to explore the histological development process of THO-TMJ. MATERIALS AND METHODS: Condylar cartilage of one-month-old male mice was partially removed from the left joint with small scissors to induce THO-TMJ. The phenotypes were observed using gross observation, microcomputed tomography (micro-CT) scans and histological examination from one month to six months after surgery. RESULTS: The micro-CT examination results showed that the injured condyle integrated with ectopic bone tissue to form an osteophyte and that the volume and density of the osteophyte grew exponentially with time. Hematoxylin and eosin (H&E), safranin O and fast green staining of the THO-TMJ specimens revealed that the ectopic bone tissue was mainly nonmineralized fibrous tissue 1 month after surgery. This tissue gradually transformed into cartilage 3 months after surgery. Finally, the tissues transformed into mature bone tissue 6 months after surgery. Immunofluorescence staining showed VEGF-α expression in the heterotopic tissue 1 month after surgery, and the expression of Sox9 in the heterotopic tissue was obvious 3 months after surgery. Furthermore, OCN expression was evident in most of the heterotopic tissue 6 months after surgery. The results also showed clear hypoxia-inducible factor 1-alpha (Hif-1α) expression in the injured chondrocytes of the condyle, especially in the articular proliferative zone and fibrocartilaginous zone. CONCLUSIONS: The THO-TMJ imaging characteristics indicated an exponential change with time. Histologically, the development process of THO-TMJ is an endochondral ossification process and includes three stages, fibroproliferative, chondrogenic and osteogenic stage. In addition, Hif-1α, which was expressed in some of the injured chondrocytes, may play an essential role in the initial THO-TMJ.

Macroalgae bloom decay decreases the sediment organic carbon sequestration potential in tropical seagrass meadows of the South China Sea.

Seagrass meadows are experiencing worldwide declines mainly because of nutrient enrichment, which always result in macroalgae bloom and consequently periodic collapse and decomposition. However, effects of macroalgae decay on the sediment organic carbon (SOC) sequestration capacity remain unknown. Depending on the macroalgae biomass in eutrophic seagrass meadows of South China Sea, we carried out a laboratory chamber experiment to investigate the sediment labile organic carbon (OC) compositions and the influencing SOC transformation enzyme activity variations of seagrass meadows in response to common macroalgae bloom species (Cladophora spp.) decomposition. Although the dehydrogenase and ß-glucosidase activities were not affected by macroalgae decomposition, the macroalgae decomposition significantly elevated the salt-extractable carbon (SEC) content, SEC/SOC, levels of invertase and polyphenol oxidase activities, and the CO2 release. Overall, this study indicates that macroalgae decomposition stimulates the SOC transformation, and therefore, it is not benefit for SOC sequestration within seagrass meadows of the South China Sea.

Antioxidant properties of a vegetable-fruit beverage fermented with two Lactobacillus plantarum strains.

Fermented vegetable-fruit beverages are a popular fermented food, with many potential health benefits. In this study, two commercial Lactobacillus plantarum strains were selected to ferment a beverage containing apples, pears, and carrots. The metabolites and antioxidant activities were examined during the fermentation process. Results showed that lactic acid and acetic acid accumulated gradually, whereas malic acid decreased. Glucose and fructose increased from 0.48 and 14.8 g/L to 7.7 and 20.8 g/L, respectively, while sucrose decreased slightly. Ascorbic acid also increased continuously during the fermentation to 90.74 mg/100 mL. DPPH and ABTS radical scavenging activity and FRAP reached their maximum value after 4-8 days. The accumulation of TPC, TFC, and SOD reached their maximum value on the 8th day of fermentation. Our study revealed that the L. plantarum-fermented vegetable-fruit beverage showed significant antioxidant activity, which is helpful in evaluating the fermentation end-point and developing a high-quality fermented beverage.

[McCune-Albright syndrome with hyperthyroidism: report of one case and review of literature].

McCune-Albright Syndrome(MAS) is a rare disease characterized by triad of polyostotic fibrous dysplasia of bone(FD), precocious puberty, and café-au-lait skin pigmentation. This paper reported a case of McCune-Albright syndrome with hyperthyroidism, and discussed the pathogenesis, clinical manifestations, diagnosis and treatment of MAS.

Successful cisplatin-etoposide chemotherapy-based treatment of a primary small cell neuroendocrine carcinoma of the tonsil with multiple metastases: a case report.

Extrapulmonary small cell neuroendocrine carcinoma (SNEC) is an extremely rare and highly malignant tumor with a poor prognosis. Multiple metastases of SNEC are even more rare, and patients with locally advanced and metastatic disease generally face a poor outcome. To date, only a few cases of SNEC have been reported. Here, we describe a rare case of a 70-year-old female patient with SNEC of the tonsil who presented with multiple metastases and had achieved a complete response (CR) of the primary lesion and cervical lymph nodes for more than 1 year after receiving palliative chemotherapy. Initially, the patient presented with a 2-month history of throat pain. Magnetic resonance imaging and computed tomography revealed a soft mass with moderate enhancement on the left tonsil, which was confirmed by incisional aspiration biopsy. She was additionally sent for a positron emission tomography scan to evaluate small metastases in the left cervical lymph node, right lung, multiple mediastinal lymph nodes, and the fourth lumbar (L4) vertebra body metastases. Histopathological examination of the SNEC confirmed a nested, typical endocrine appearance with small round cells containing ovoid-shaped nuclei and high mitotic activity. Immunohistochemically, the tumor cells were positive for cytokeratin 8/18+, synaptophysin+, CD56+, and Ki-67 (<50%). The patient received 6 cycles of cisplatin combined with etoposide and was subsequently placed under close observation (<12 months). To date, she has achieved a CR of the primary lesion and cervical lymph nodes. In summary, we have described a case of successful treatment after chemotherapy for SNEC and have elucidated professional knowledge regarding the relevant aspects of SNEC.

Increasing thermal stability of glutamate decarboxylase from Escherichia. coli by site-directed saturation mutagenesis and its application in GABA production.

Gamma-amino butyric acid (GABA) is an important bio-product used in pharmaceuticals, functional foods, and a precursor of the biodegradable plastic polyamide 4 (Nylon 4). Glutamate decarboxylase B (GadB) from Escherichia. coli is a highly active biocatalyst that can convert l-glutamate to GABA. However, its practical application is limited by the poor thermostability and only active under acidic conditions of GadB. In this study, we performed site-directed saturation mutagenesis of the N-terminal residues of GadB from Escherichia coli to improve its thermostability. A triple mutant (M6, Gln5Ile/Val6Asp/Thr7Gln) showed higher thermostability, with a 5.6 times (560%) increase in half-life value at 45 °C, 8.7 °C rise in melting temperature (Tm) and a 14.3 °C rise in the temperature at which 50% of the initial activity remained after 15 min incubation (T1550), compared to wild-type enzyme. Protein 3D structure analysis showed that the induced new hydrogen bonds in the same polypeptide chain or between polypeptide chains in E. coli GadB homo-hexamer may be responsible for the improved thermostability. Increased thermostability contributed to increased GABA conversion ability. After 12 h conversion of 3 mol/L l-glutamate, GABA produced and mole conversion rate catalyzed by M6 whole cells was 297 g/L and 95%, respectively, while those by wild-type GAD was 273.5 g/L and 86.2%, respectively.

[ISOLATION OF RAT PATELLAR TENDON STEM CELLS AND EFFECT OF MECHANICAL STRETCHING ON Sox-9 EXPRESSION].

OBJECTIVE: To isolate the tendon stem cells (TSCs) from rat patellar tendon and to investigate the effect of mechanical stretching on the expression of Sox-9. METHODS: TSCs were isolated from Sprague Dawley rat (12 weeks old) patellar tendon by collagenase digestion and low density culture. The cell colony morphology and number were observed by crystal violet staining; the cell morphology was observed by inverted phase contrast microscope, and the immunophenotypes of mesenchymal stem cells (MSCs) were determined by flow cytometry. The TSCs at passage 3 was given the mechanical stretching at 4%, 0.17 Hz for 4 hours and 24 hours in the experimental group, and cells without stretching was used as control. The Sox-9 gene and protein expressions were detected by real-time fluorescence quantitative PCR and Western blot. RESULTS: Primary cells showed clonal growth and star shape; after subculture, cells at passage 1 showed fibroblast-like shape. The cells formed cell colonies after 7 days; the expressions were positive for CD29, CD44, and CD90 and negative for CD45. The result of real-time fluorescence quantitative PCR showed that Sox-9 gene was down-regulated at 4 hours after mechanical stretching compared with control (P < 0.05), and up-regulated at 24 hours after mechanical stretching when compared with control group (P < 0.05). The result of Western blot showed that Sox-9 protein expression was lower at 4 hours after stretching, but higher at 24 hours after mechanical stretching than that in control group (P < 0.05). CONCLUSION: The rat patellar TSCs can be isolated successfully, and mechanical stretching inhibits the Sox-9 expression, but the inhibited effect might stimulate the Sox-9 expression after the mechanical stretching effect disappears.