Biphasic photobiomodulation of inflammation in mouse models of common wounds, infected wounds, and diabetic wounds.

Bidirectional photobiomodulation (PBM) therapy is an active research area. However, most studies have focused on its dependence on optical parameters rather than on its tissue-dependent effects. We constructed mouse models of wounds in three inflammatory states (normal, low, and high levels of inflammations) to assess the bidirectional regulatory effect of PBM on inflammation. Mice were divided into three groups to prepare common wounds, diabetic wounds, and bacteria-infected wounds. The same PBM protocol was used to regularly irradiate the wounds over a 14 d period. PBM promoted healing of all three kinds of wounds, but the inflammatory manifestations in each were significantly different. In common wounds, PBM slightly increased the aggregation of inflammatory cells and expression of IL-6 but had no effect on the inflammatory score. For wounds in a high level of inflammation caused by infection, PBM significantly increased TNF-α expression in the first 3 d of treatment but quickly eliminated inflammation after the acute phase. For the diabetic wounds in a low level of inflammation, PBM intervention significantly increased inflammation scores and prevented neutrophils from falling below baseline levels at the end of the 14 d observation period. Under fixed optical conditions, PBM has a bidirectional (pro- or anti-inflammatory) effect on inflammation, depending on the immune state of the target organism and the presence of inflammatory stimulants. Our results provide a basis for the formulation of clinical guidelines for PBM application.

Construction and assessment of a joint prediction model and nomogram for colorectal cancer.

Background: Colorectal cancer (CRC) is one of the most common tumors in the digestive system, and all its risk factors are not yet known. It is important to identify valuable clinical indicators to predict the risk of CRC. Methods: A total of 227 participants, comprising 162 healthy adults and 65 patients diagnosed with CRC at Tianjin Hospital from January 2017 to March 2022, were included in this study. Electrochemiluminescence was adopted to test the expression levels of carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA199). Univariate and multivariate logistic regression analyses were performed to identify independent risk factors for CRC, and a joint prediction model was then constructed. A nomogram was prepared, and the model was later assessed using the receiver operating characteristic curve and calibration curve. Results: The univariate analysis showed that there were statistically significant differences between the two groups in terms of smoking (χ2=8.67), fecal occult blood (χ2=119.41), Helicobacter pylori (H. pylori) infection (χ2=30.87), a history of appendectomy (χ2=5.47), serum total bile acid levels (t=19.80), serum CEA levels (t=37.82), serum CA199 levels (t=6.82), and serum ferritin levels (t=54.31) (all P<0.05). The multiple logistic regression analysis showed that smoking, fecal occult blood, H. pylori infection, a history of appendectomy, serum CEA levels, and serum CA199 levels were independent risk factors for CRC (all P<0.05). Based on the above findings, a joint prediction model was constructed, and the area under the receiver operator characteristic (ROC) curve of the model was 0.842. A nomogram and calibration curve was drawn, and the internal validation results indicated that the model had good diagnostic value. Conclusions: Smoking, fecal occult blood, H. pylori infection, a history of appendectomy, serum CEA levels, and serum CA199 levels are independent risk factors for CRC, and the prediction model based on these factors had good predictive ability.

Bioinformatics analysis of Myelin Transcription Factor 1.

BACKGROUND AND OBJECTIVE: We aimed to further study the role of Myelin Transcription Factor 1(MyT1) in tumor and other diseases and epigenetic regulation, and better understand the regulatory mechanism of MyT1. METHODS: Using bioinformatics analysis, the structure and function of MyT1sequence were predicted and analyzed using bioinformatics analysis, and providing a theoretical basis for further experimental verification and understanding the regulatory mechanism of MyT1. The first, second and third-level structures of MyT1 were predicted and analyzed by bioinformatics analysis tools. RESULTS: MyT1 is found to be an unstable hydrophilic protein, rather than a secretory protein, with no signal peptide or trans-membrane domain; total amino acids located on the surface of the cell membrane. It contains seven zinc finger domains structurally. At sub-cellular level, MyT1 is localized in the nucleus. The phosphorylation site mainly exists in serine, and its secondary structure is mainly composed of random coils and alpha helices; the three-dimensional structure is analyzed by modeling. CONCLUSIONS: In this study, the structure and function of MyT1 protein were predicted, thereby providing a basis for subsequent expression analysis and functional research; it laid the foundation for further investigation of the molecular mechanism involved in the development of diseases.

Characterization of microvessels and parenchyma in in-line phase contrast imaging CT: healthy liver, cirrhosis and hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is a cancer with a poor prognosis, and approximately 80% of HCC cases develop from cirrhosis. Imaging techniques in the clinic seem to be insufficient for revealing the microstructures of liver disease. In recent years, phase contrast imaging CT (PCI-CT) has opened new avenues for biomedical applications owing to its unprecedented spatial and contrast resolution. The aim of this study was to present three-dimensional (3D) visualization of human healthy liver, cirrhosis and HCC using a PCI-CT technique called in-line phase contrast imaging CT (ILPCI-CT) and to quantitatively evaluate the variations of these tissues, focusing on the liver parenchyma and microvasculature. METHODS: Tissue samples from 9 surgical specimens of normal liver (n=3), cirrhotic liver (n=2), and HCC (n=4) were imaged using ILPCI-CT at the Shanghai Synchrotron Radiation Facility (SSRF) without contrast agents. 3D visualization of all ex vivo liver samples are presented. To quantitatively evaluate the vessel features, the vessel branch angles of each sample were clearly depicted. Additionally, radiomic features of the liver parenchyma extracted from the 3D images were measured. To evaluate the stability of the features, the percent coefficient of variation (%COV) was calculated for each radiomic feature. A %COV <30 was considered to be low variation. Finally, one-way ANOVA, followed by Tukey's test, was used to determine significant changes among the different liver specimens. RESULTS: ILPCI-CT allows for a clearer view of the architecture of the vessels and reveals more structural details than does conventional radiography. Combined with the 3D visualization technique, ILPCI-CT enables the acquisition of an accurate description of the 3D vessel morphology in liver samples. Qualitative descriptions and quantitative assessment of microvessels demonstrated clear differences among human healthy liver, cirrhotic liver and HCC. In total, 38 (approximately 51%) radiomic features had low variation, including 11 first-order features, 16 GLCM features, 6 GLRLM features and 5 GLSZM features. The differences in the mean vessel branch angles and 3 radiomic features (first-order entropy, GLCM-inverse variance and GLCM-sum entropy) were statistically significant among the three groups of samples. CONCLUSIONS: ILPCI-CT may allow for morphologic descriptions and quantitative evaluation of vessel microstructures and parenchyma in human healthy liver, cirrhotic liver and HCC. Vessel branch angles and radiomic features extracted from liver parenchyma images can be used to distinguish the three kinds of liver tissues.

Metronomic photodynamic therapy with 5-aminolevulinic acid induces apoptosis and autophagy in human SW837 colorectal cancer cells.

Metronomic photodynamic therapy (mPDT) has emerged as an attractive treatment for the selective destruction of tumor cells by induction of apoptosis. Here, we compared the effects of mPDT and acute photodynamic therapy (aPDT) on human SW837 colorectal cancer (CRC) cells. CRC cells were subjected to mPDT using various exposure durations, concentrations of 5-aminolevulinic acid (ALA), fluence rates and energy densities. The effects were compared with those induced by aPDT. We found that apoptosis and autophagy were earlier induced to a greater extent by mPDT than by the same dose applied as aPDT. The survival rates for mPDT vs. aPDT were 35.2%, 32.4%,27.6%,31.6% vs. 85.7%, 71.1%, 67.8%, 42.1% after 3, 6, 12, and 24 h PDT, respectively. For the same time points, the apoptotic rates for mPDT vs. aPDT were 43.2%, 47.3%, 54.7%, and 50.3% vs. 14.6%, 17.6%, 27.1%, and 53.2%, respectively. mPDT induced a peak rate of autophagy of 20.0% at 3 h, whereas aPDT induced two smaller peaks at 3 h (14.1%) and 12 h (15.8%). Advanced autophagosomes were more abundant in mPDT- than aPDT-treated cells and appeared earlier after mPDT (3 h) than after aPDT (3-12 h). Western Bloting results showed that the ratio of LC3B-II/ß - actin at 3 h was higher (1.04 times) after mPDT than aPDT. Collectively, these datas indicated that ALA-mPDT was more effective than the same dose of ALA-aPDT at inducing SW837 CRC cell death via apoptosis and autophagy. Thus, mPDT may be a superior choice than aPDT for the treatment of human CRC.

Study on Umbilical Cord-Matrix Stem Cells Transplantation for Treatment of Acute Traumatic Brain Injury in Rats.

AIM: To evaluate the effect of umbilical cord derived mesenchymal stem cells (UC-MSCs) transplantation on traumatic brain injury (TBI). MATERIAL AND METHODS: UC-MSCs were isolated from human umbilical cord and TBI rat model was constructed. 30 male SD rats were randomly divided into 3 groups: control group, TBI group and MSCs transplantation group. Rats in MSCs group received the injection of a total of 1.5 C- 106 MSCs (25 I»l) via ventricle at operated ventricular coordinates (0 at bregma, 1.5 mm at lateral, 1.1 mm at behind, 4.5 mm in depth). RESULTS: 80% confluence of cells was formed from tissue at day 10 and the amount of CD90, CD73, CD105 positive cells increased correspondingly. In TBI model, clear hyperemia, edema and obvious infiltration of inflammatory cells in brain tissue were found. However, the manifestations were alleviated after the treatment of MSCs. In MSCs group, GFP in the brain tissue and the area around the vessels were found after the injection, while the expression levels of micro-vessel density (MVD), brain-derived neurotrophic factor (BDNF) and glial fibrillary acidic protein (GFAP) were elevated. CONCLUSION: UC-MSCs transplantation for treatment of acute TBI could effectively reduce the injury and improve the vascular reconstruction.

Mechanism of interaction between ocular and nasal neurogenic inflammation in allergic rhinoconjunctivitis.

PURPOSE: The mechanisms of naso-ocular interaction in allergic rhinoconjunctivitis are not well understood. Neurogenic inflammation affects both eyes and nose via the same neurogenic factors. The purpose of this study was to investigate the effects of neurogenic inflammation on conjunctival inflammation following nasal allergen provocation. METHODS: Sensitized rats were exposed to ovalbumin (OVA) via the nose. Parts of the nasal mucosa and conjunctivae were sliced and used for hematoxylin-eosin staining, immunohistochemical analysis, western blotting, and real-time polymerase chain reaction. The slides were observed under a light microscope, and the acquired images were analyzed. The levels of substance P (SP), vasoactive intestinal peptide (VIP), and nerve growth factor (NGF) were detected. RESULTS: The levels of SP, VIP, and NGF were increased in both nasal mucosa and conjunctivae 1 h and 24 h after OVA administration (p < 0.05). Higher levels of SP, VIP, and NGF expression were observed in the nasal mucosa and conjunctivae 24 h after OVA administration (p < 0.05). Following damage of the nasal sensory nerves by capsaicin, the protein and mRNA levels of SP, VIP, and NGF were reduced. CONCLUSION: In conclusion, the increased levels of VIP, SP, and NGF might be responsible for the ocular reaction following nasal challenge with allergen in rats.

Effect of photobiomodulation on neural differentiation of human umbilical cord mesenchymal stem cells.

Photobiomodulation therapy (PBMT) can enhance the mesenchymal stem cell (MSC) proliferation, differentiation, and tissue repair and can therefore be used in regenerative medicine. The objective of this study is to investigate the effects of photobiomodulation on the directional neural differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) and provide a theoretical basis for neurogenesis. hUC-MSCs were divided into control, inducer, laser, and lasers combined with inducer groups. A 635-nm laser and an 808-nm laser delivering energy densities from 0 to 10 J/cm2 were used in the study. Normal cerebrospinal fluid (CSF) and injured cerebrospinal fluid (iCSF) were used as inducers. The groups were continuously induced for 3 days. Cellular proliferation was evaluated using MTT. The marker proteins nestin (marker protein of the neural precursor cells), NeuN (marker protein of neuron), and GFAP (glial fibrillary acidic protein, marker proteins of glial cells) were detected by immunofluorescence and western blot. We found that irradiation with 635-nm laser increased cell proliferation, and that with 808 nm laser by itself and combined with cerebrospinal fluid treatment generated significant neuron-like morphological changes in the cells at 72 h. Nestin showed high positive expression at 24 h in the 808 nm group. The expression of GFAP increased in the 808-nm combined inducer group at 24 h but decreased at 72 h. The expression of neuN protein increased only at 72 h in both the 808-nm combined inducer group and inducer group. We concluded that 808 nm laser irradiation could help CSF to induce neuronal differentiation of hUC-MSCs in early stage and tend to change to neuron rather than glial cells.

Effect of 405 nm low intensity irradiation on the absorption spectrum of in-vitro hyperlipidemia blood.

BACKGROUND: Laser therapy is reported to be clinically effective for improving microcirculation, rheological properties and blood lipid profiles despite the lack of certainty on the mechanism. OBJECTIVE: This study intends to provide methods to drop blood lipid level of hyperlipidemia samples by low-intensity laser irradiation therapy and provide reasoning of mechanism. METHODS: Twenty whole blood samples of high level of lipids profile are irradiated by 405 nm low-intensity laser at 12 J/cm2 twice a day for 3 days and compared with normal lipids profile group. Then whole blood sample are centrifuged to obtain result of erythrocyte for further interpretation. Multi-scan spectrum microplate reader is used to measure absorption spectrum and data is analyzed by software SPSS 14.0. RESULTS: Results show that after 405 nm low-intensity laser irradiation, whole blood samples of high lipid level statistically have higher absorbance peak value than normal samples while erythrocyte samples have lower absorbance peak value. CONCLUSIONS: From the divergence of absorption peak value change after low-intensity laser irradiation for whole blood sample and erythrocyte, we suspect that low level laser irradiation affects the enzymes activity of lipid metabolism, improves the cholesterol balance of plasma and cytoplasm in erythrocyte, and decreases aggregation of the erythrocyte.

Umbilical cord mesenchymal stem cell (UC-MSC) transplantations for cerebral palsy.

This study reports a case of a 4-year-old boy patient with abnormalities of muscle tone, movement and motor skills, as well as unstable gait leading to frequent falls. The results of the electroencephalogram (EEG) indicate moderately abnormal EEG, accompanied by irregular seizures. Based on these clinical characteristics, the patient was diagnosed with cerebral palsy (CP) in our hospital. In this study, the patient was treated with umbilical cord mesenchymal stem cell (UC-MSC) transplantation therapy. This patient received UC-MSC transplantation 3 times (5.3*107) in total. After three successive cell transplantations, the patient recovered well and showed obvious improvements in EEG and limb strength, motor function, and language expression. However, the improvement in intelligence quotient (IQ) was less obvious. These results indicate that UC-MSC transplantation is a promising treatment for cerebral palsy.

High-voltage pulsed electric field plus photodynamic therapy kills breast cancer cells by triggering apoptosis.

This study evaluated the effects and mechanism of action of combining irreversible electroporation (IRE) and photodynamic therapy (PDT) in breast cancer cells in vitro and in vivo. Jin's formula was used to assess killing efficacy of different IRE+PDT dosing combinations in breast cancer MCF-7 cells. Flow cytometry, high-content imaging, and confocal laser scanning microscopy were used to detect apoptosis. qRT-PCR and western blotting were used to evaluate expression of apoptosis-related genes and proteins. IRE+PDT combination therapy was administered to BALB/C mice with breast cancer tumors in vivo; tumor size was used to assess treatment efficacy. Killing mechanisms were examined using transmission electron microscopy and immunohistochemistry. We found that IRE+PDT combination therapy produced significant synergistic killing effects in breast cancer cells (highest Jin q value of 1.32). Early apoptosis rates were significantly higher in the IRE+PDT group (16.0%) than in IRE-alone (7.6%) and PDT-alone (4.6%) groups (P<0.05). qRT-PCR showed higher Caspase-1, -3, -5, -6, -7, -8, and -9 and TNFRSF1A expression with IRE+PDT than with control. Western blots showed increased cleaved Caspase-3, -7, and -9, and PARP levels in the IRE+PDT group. In vivo tumor suppression rate for IRE (1200 V)+PDT (10 mg/kg) was 68.3%. Combination therapy produced the most obvious apoptosis effects. Compared with controls, the IRE+PDT group exhibited lower new blood vessel (VEGF, CD31), metastasis (TGF-ß), and cell proliferation (Ki-67) indicators and higher inflammation indicator (TNF-α) 1 day post-treatment. Thus, combining IRE and PDT enhanced their anti-tumor effects in breast cancer, and apoptosis played a key role in this process.

Hypothermia-Modulating Matrix Elasticity of Injured Brain Promoted Neural Lineage Specification of Mesenchymal Stem Cells.

Both chemical and physical microenvironments appear to be important for lineage specification of umbilical cord mesenchymal stem cells (UCMSCs). However, physical factors such as the elastic modulus in traumatic brain injury (TBI) are seldom studied. Intracranial hypertension and cerebral edema after TBI may change the brain's physical microenvironment, which inhibits neural lineage specification of transplanted UCMSCs. The purpose of this study is to investigate the potential regulatory effect of mild hypothermia on the elastic modulus of the injured brain. First, we found that more UCMSCs grown on gels mimicking the elastic modulus of the brain (0.5 kPa) differentiated into neural cells, which were verified with the formation of branched cells and the expression of neural markers. Then, UCMSCs were transplanted into TBI rats, and we observed that mild hypothermia resulted in the differentiation of more neurons and astrocytes from transplanted UCMSCs. To demonstrate that more neural specification of UCMSCs was due to the regulation of the elastic modulus, we monitored intracranial pressure and cerebral edema. The results showed that mild hypothermia significantly reduced intracranial pressure and brain water content, indicating modulation of the elastic modulus by mild hypothermia. An examination with atomic force microscopy (AFM) in a cell injury model in vitro further verified hypothermia-regulated elastic modulus. In this study, we found a novel role of mild hypothermia in modulating the elastic modulus of the injured brain, resulting in the promotion of neural lineage specification of UCMSCs, which suggested that the combination of mild hypothermia had more advantages in cell-based therapy after TBI.

The Distribution of Transplanted Umbilical Cord Mesenchymal Stem Cells in Large Blood Vessel of Experimental Design With Traumatic Brain Injury.

The authors aim to track the distribution of human umbilical cord mesenchymal stem cells (MSCs) in large blood vessel of traumatic brain injury -rats through immunohistochemical method and small animal imaging system. After green fluorescent protein (GFP) gene was transfected into 293T cell, virus was packaged and MSCs were transfected. Mesenchymal stem cells containing GFP were transplanted into brain ventricle of rats when the infection rate reaches 95%. The immunohistochemical and small animal imaging system was used to detect the distribution of MSCs in large blood vessels of rats. Mesenchymal stem cells could be observed in large vessels with positive GFP expression 10 days after transplantation, while control groups (normal group and traumatic brain injury group) have negative GFP expression. The vascular endothelial growth factor in transplantation group was higher than that in control groups. The in vivo imaging showed obvious distribution of MSCs in the blood vessels of rats, while no MSCs could be seen in control groups. The intravascular migration and homing of MSCs could be seen in rats received MSCs transplantation, and new angiogenesis could be seen in MSCs-transplanted blood vessels.

The research progress of mesenchymal stem cells in the treatment of Traumatic brain injury.

Traumatic brain injury (TBI) is the leading cause of mortality and morbidity in children and adults throughout the world. It is urgent to ameliorate TBI damage and reduce the disability and case-fatality rate. Stem cell therapy is another medical revolution after drug and surgical medication. Mesenchymal stem cells (MSCs) are a class of cells with significant self-renewal and multi-lineage differentiation properties and be favorable for the treatment of various diseases and injuries, it could be envisioned that MSCs transplantation may be a promising treatment for TBI. Currently, stem cell therapy has shown promising effects in the treatment of many diseases. In this article, we will review the characteristics of MSCs, MSCs for neuronal function restoration, the properties of immune-modulatory of MSCs, and the anti-apoptotic effects of MSCs, the angiogenesis effect of MSCs,and the safety issues in MSCs therapy in TBI.

[Effects of human umbilical cord mesenchymal stem cells on vascular endothelial growth factor and IL-6 expression in tissue of AMI rats].

OBJECTIVE: To investigate the effects of human umbilical cord mesenchymal stem cells (UC-MSCs) on vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) expression in acute myocardium infarction (AMI) rats. METHODS: The human UC-MSCs were cultured to the 4th generation for experiment. Sixty male Sprague-Dawley (SD) rats were randomly divided into sham group, AMI model group and UC-MSCs group, with 20 in each group. AMI animal model was produced by ligation of anterior descending coronary artery; in the sham group, the threading vein was gone below without ligation. In UC-MSCs group 2×106 UC-MSCs were infused through the caudal vein at 24 hours after successful model production. The animals were sacrificed after 7 days; the myocardial tissue and coronary artery below the ligation line were harvested. The mRNA and protein expressions of IL-6 in myocardium were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot. The positive expression of VEGF in coronary artery was observed by immunohistochemisty. RESULTS: Compared with the sham group, the mRNA and protein expressions of IL-6 in myocardium in AMI model group were increased significantly (gray value: 0.732±0.131 vs. 0.321±0.080, 0.678±0.191 vs. 0.286±0.061, both P < 0.05). Compared with the AMI model group, the mRNA and protein expressions of IL-6 in myocardium in UC-MSCs group were decreased significantly (gray value: 0.300±0.104 vs. 0.732±0.131, 0.312±0.101 vs. 0.678±0.191, both P < 0.05). Observation under light microscope, the VEGF positive cells in AMI model group was increased significantly compared with the sham group (cells/HP: 21.1±2.2 vs. 7.6±1.3, P < 0.05), the VEGF positive cells in UC-MSCs group were increased significantly compared with the AMI model group (cells/HP: 41.5±3.1 vs. 21.1±2.2, P < 0.05). CONCLUSIONS: Human UC-MSCs could promote angiogenesis by the improvement of VEGF in coronary artery and inhibit the inflammation by the reduction of IL-6 in rats with AMI.

Establishment of an ideal time window model in hypothermic-targeted temperature management after traumatic brain injury in rats.

Although hypothermic-targeted temperature management (HTTM) holds great potential for the treatment of traumatic brain injury (TBI), translation of the efficacy of hypothermia from animal models to TBI patientshas no entire consistency. This study aimed to find an ideal time window model in experimental rats which was more in accordance with clinical practice through the delayed HTTM intervention. Sprague-Dawley rats were subjected to unilateral cortical contusion injury and received therapeutic hypothermia at 15mins, 2 h, 4 h respectively after TBI. The neurological function was evaluated with the modified neurological severity score and Morris water maze test. The brain edema and morphological changes were measured with the water content and H&E staining. Brain sections were immunostained with antibodies against DCX (a neuroblast marker) and GFAP (an astrocyte marker). The apoptosis levels in the ipsilateral hippocampi and cortex were examined with antibodies against the apoptotic proteins Bcl-2, Bax, and cleaved caspase-3 by the immunofluorescence and western blotting. The results indicated that each hypothermia therapy group could improve neurobehavioral and cognitive function, alleviate brain edema and reduce inflammation. Furthermore, we observed that therapeutic hypothermia increased DCX expression, decreased GFAP expression, upregulated Bcl-2 expression and downregulated Bax and cleaved Caspase-3 expression. The above results suggested that HTTM at 2h or even at 4h post-injury revealed beneficial brain protection similarly, despite the best effect at 15min post-injury. These findings may provide relatively ideal time window models, further making the following experimental results more credible and persuasive.