Effect and mechanism of Zuogui Pills on neural function recovery in ischemic stroke mice based on OPN/IGF-1/mTOR / 中国中药杂志

To explore the effect and mechanism of Zuogui Pills in promoting neural tissue recovery and functional recovery in mice with ischemic stroke. Male C57BL/6J mice were randomly divided into a sham group, a model group, and low-, medium, and high-dose Zuogui Pills groups(3.5, 7, and 14 g·kg~(-1)), with 15 mice in each group. The ischemic stroke model was established using photochemical embolization. Stiker remove and irregular ladder walking behavioral tests were conducted before modeling and on days 7, 14, 21, and 28 after medication. Triphenyl tetrazolium chloride(TTC) staining was performed on day 3 after modeling, and T2-weighted imaging(T2WI) and diffusion-weighted imaging(DWI) were performed on day 28 after medication to evaluate the extent of brain injury. Hematoxylin-eosin(HE) staining was performed to observe the histology of the cerebral cortex. Axonal marker proteins myelin basic protein(MBP), growth-associated protein 43(GAP43), mammalian target of rapamycin(mTOR), and its downstream phosphorylated s6 ribosomal protein(p-S6), as well as mechanism-related proteins osteopontin(OPN) and insulin-like growth factor 1(IGF-1), were detected using immunofluorescence and Western blot. Zuogui Pills had a certain restorative effect on the neural function impairment caused by ischemic stroke in mice. TTC staining showed white infarct foci in the sensory-motor cortex area, and T2WI imaging revealed cystic necrosis in the sensory-motor cortex area. The Zuogui Pills groups showed less brain tissue damage, fewer scars, and more capillaries. The number of neuronal axons in those groups was higher than that in the model group, and neuronal activity was stronger. The expression of GAP43, OPN, IGF-1, and mTOR proteins in the Zuogui Pills groups was higher than that in the model group. In summary, Zuogui Pills can promote the recovery of neural function and axonal growth in mice with ischemic stroke, and its mechanism may be related to the activation of the OPN/IGF-1/mTOR signaling pathway.

Mechanism of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma Extracts in Delaying High Glucose-induced Vascular Calcification in Mice / 中国实验方剂学杂志

Objective::To investigate the protective effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts on vascular calcification induced by high glucose in mice by observing the expression of osteopontin (OPN) and smooth muscle 22α (SM22α) as well as vascular calcium deposition in the common carotid artery and thoracic aorta of mice. Method::Totally 130 male C57BL/6 mice were randomly divided into normal control group and high glucose group. The mice in high glucose group were intraperitoneally injected with streptozotocin(STZ), and fed on a high-fat diet for 7 months. Then, the mice were randomly divided into model group, low-dose and high-dose Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts groups (0.819, 1.638 g·kg-1), and metformin group (150 mg·kg-1). Each group was intragastrically administered once a day for 9 weeks. The changes in blood glucose were measured. Seven days before the end of the administration, a group of 4-week old male C57BL/6 mice were purchased and fed normally for one week as a youth group. At the end of the administration, the common carotid artery and thoracic aorta tissues of the mice were collected. Von Kossa staining was used to determine the degree of calcium deposition in the common carotid artery and thoracic aorta. The expression levels of OPN and SM22α protein in the common carotid artery and thoracic aorta were detected by immunohistochemistry. The expression of OPN and SM22α protein in the common carotid artery of mice was determined by Western blot. Result::As compared with the young group, the blood glucose of the normal control group was slightly increased without statistical difference, the common carotid artery and thoracic aorta were uniformly stained, and no black granular precipitate was observed. As compared with the normal control group, the blood glucose of the model group was increased (P<0.01), with a large amount of brown-black particles deposited in the intimal elastic fibers, showing obvious calcium salt deposition. As compared with the model group, blood glucose was significantly decreased in each administration group (P<0.05, P<0.01), and the degree of vascular calcium salt deposition was significantly reduced. There were no significant changes in expression levels of OPN protein and SM22α protein in the common carotid artery and thoracic aorta between the youth group and normal control group. As compared with the normal control group, the expression of intimal OPN protein in the common carotid artery and thoracic aorta of the model group was positive, SM22α protein expression was weakly positive, and the gray value of OPN protein expression in the common carotid artery was significantly increased (P<0.01), while the gray value of SM22α protein was decreased significantly (P<0.01). As compared with the model group, the expression levels of intimal OPN protein and SM22α protein in the common carotid artery and thoracic aorta of each administration group were significantly improved, and the gray value of OPN protein expression in the common carotid artery was reduced (P<0.05, P<0.01), while SM22α protein expression was significantly increased (P<0.01). Conclusion::High glucose can induce calcification of common carotid artery and thoracic aorta in mice and accelerate vascular aging. This formation process may be related to the expression of OPN and SM22α. Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma extracts can reduce vascular calcification and delay vascular aging by regulating the expression of OPN and SM22α.

The diagnostic and prognostic value of osteopontin in patients with pancreatic cancer: A meta-analysis / 中华肝胆外科杂志

Objective To study the expression of osteopontin (OPN) in pancreatic cancer and to determine its prognostic significance.Methods Studies which evaluated the relationship between the expression of OPN and pancreatic cancer published up to May 2018 were found by searching the electronic databases which included the PubMed,Embase,Cochrane Library,CNKI,Wanfang and VIP.The RevMan 5.3 was used to analyze the data in this meta-analysis by determining the odds ratios (ORs) and their 95％ confidence intervals (CIs).Results Using our predetermined selection criteria,this meta-analysis included 13 studies,with 1073 cases of pancreatic cancer tissues in the case group and 346 cases of normal tissues in the control group.The results indicated OPN was over expressed in pancreatic cancers (OR =8.13,95％ CI:6.00 ～ 11.02;P ＜ 0.05).The differences in the expressions of OPN between the well differentiation group and the moderate-poor differentiation group (OR =0.37,95％ CI:O.25 ～ 0.57;P ＜ 0.05),between the clinical stage Ⅰ-Ⅱ group and the clinical stage Ⅲ-Ⅳ group (OR =0.23,95 ％ CI:0.14 ～ 0.38;P ＜0.05),and between the lymph node metastasis group and the no lymph node metastasis group (OR =3.77,95％ CI:1.99 ～ 7.13;P ＜ 0.05) were significant.The expressions of OPN did not correlate with gender,age and location of pancreatic cancer.Conclusions Current evidence indicated that OPN was significantly correlated with pancreatic cancer and its clinicopathological features.OPN could be used as an effective diagnostic and survival evaluation marker.

Post-surgical resection prognostic value of combined OPN, MMP7, and PSG9 plasma biomarkers in hepatocellular carcinoma / 医学前沿

Biomarkers for hepatocellular carcinoma (HCC) following curative resection are not currently sufficient for prognostic indication of overall survival (OS) and disease-free survival (DFS). The aim of this study was to investigate the prognostic performance of osteopontin (OPN), matrix metalloproteinase 7 (MMP7), and pregnancy specific glycoprotein 9 (PSG9) in patients with HCC. A total of 179 prospective patients with HCC provided plasma before hepatectomy. Plasma OPN, MMP7, and PSG9 levels were determined by enzyme-linked immunosorbent assay. Correlations between plasma levels, clinical parameters, and outcomes (OS and DFS) were overall analyzed. High OPN ( ⩾ 149.97 ng/mL), MMP7 ( ⩾ 2.28 ng/mL), and PSG9 ( ⩾ 45.59 ng/mL) were prognostic indicators of reduced OS (P < 0.001, P < 0.001, and P = 0.007, respectively). Plasma PSG9 protein level was an independent factor in predicting OS (P = 0.008) and DFS (P = 0.038). Plasma OPN + MMP7 + PSG9 elevation in combination was a prognostic factor for OS (P < 0.001). OPN was demonstrated to be a risk factorassociated OS in stage I patients with HCC and patients with low α-fetoprotein levels ( < 20 ng/mL). These findings suggested that OPN, MMP7, PSG9 and their combined panels may be useful for aiding in tumor recurrence and mortality risk prediction of patients with HCC, particularly in the early stage of HCC carcinogenesis.

Effect of Osteopontin on Nuclear Mechanics of Bone Marrow Mesenchymal Stem Cells and Its Involved Molecular Mechanisms / 医用生物力学

Objective To study the effects of osteopontin (OPN) on the nuclear mechanics of bone marrow-derived mesenchymal stem cells (BMSCs) as well as its involved mechanisms. Methods The BMSC migration was evaluated using the Transwell assay. An atomic force microscope (AFM) was used to determine the elastic modulus of the BMSC nucleus and analyze the changes in the nuclear mechanics of the BMSCs after treatment with OPN. The activation of focal adhesion kinase (FAK) and extracellular signal-regulated kinase1/2 (ERK1/2) was measured by Western blot. The role of the FAK-ERK1/2 signaling pathway in mediating the OPN-affected BMSC nuclear mechanics was investigated by employing a specific inhibitor. RT-PCR and Western blot were used to detect the expression of Lamin A/C at mRNA and protein levels in the BMSCs, respectively. Results The elastic modulus of the BMSC nucleus exhibited a significant decrease after OPN treatment compared with that of the control group. OPN could upregulate the phosphorylation level of FAK and ERK1/2, but the inhibitor of FAK or ERK1/2 restored the OPN-decreased elastic modulus of the BMSC nucleus and inhibited the BMSC migration significantly. After treatment with OPN, the expression of Lamin A/C in the BMSCs reduced significantly, and such a reduced expression could be suppressed by the inhibitor of FAK or ERK1/2. Conclusions OPN could probably downregulate the expression of Lamin A/C of the BMSCs via the FAK-ERK1/2 signaling pathway, decrease the stiffness of the BMSC nucleus, and promote the migration of the BMSCs. The research outcomes provide the experimental evidence for further understanding the mechanism of the OPN-regulated BMSC migration and its potential clinical application.

Role of miR-27a in the osteogenic differentiation of beagle maxillary sinus membrane stem cells / 口腔疾病防治

Objective@#To detect the expression level of miR-27a during the osteogenic differentiation of beagle maxillary sinus membrane stem cells (MSMSCs) and explore the role of miR-27a in the osteogenic differentiation of MSMSCs.@*Methods@#Beagle MSMSCs were cultured in vitro. The expression level of miR-27a was detected via RT-PCR after an osteogenic inductive culture was prepared. The mRNA expression levels of Runx2 and OPN were examined via RT-PCR, and the protein expression levels of Runx2 and OPN were examined via Western blot after the cells were transfected with pre-miR-27a or anti-miR-27a. Finally, osteoprogenitor cells transfected with pre-miR-27a were composited with Bio-Oss particles and subcutaneously implanted into nude mice to form ectopic bone formation models, and then the inhibition of bone formation from miR-27a was observed in vivo. @*Results@#The expression level of miR-27a in the beagle MSMSCs decreased after osteogenic inductive culturing. The relative miR-27a levels were significantly decreased at day 1 (t=3.795, P=0.023), day 3 (t=4.493, P=0.011), day 7 (t=11.591, P < 0.001), day 14 (t=12.542, P < 0.001), and day 21 (t=5.621, P=0.008) compared with day 0. In addition, the expression levels of Runx2 mRNA (t=4.923, P=0.007) and protein (t=4.425, P=0.008) were reduced after the cells were transfected with pre-miR-27a. The expression levels OPN mRNA (t=5.253, P=0.006) and protein (t=5.132, P=0.006) were also reduced. In contrast, the mRNA expression levels of Runx2 (t=3.925, P=0.013) and OPN (t=3.712, P=0.019) were increased after the cells were transfected with anti-miR-27a, and bone formation was observed after the subcutaneous implantation of beagle MSMSCs composited with Bio-Oss in nude mice. Nevertheless, ectopic bone formation was inhibited by pre-miR-27a-transfected beagle MSMSCs composited with Bio-Oss (t=7.219, P=0.0020). @* Conclusion @# MiR-27a negatively regulates the osteogenic differentiation of MSMSCs.

Correlation Analysis of Wnt/β-catenin Signaling Pathway, OPN and MMP-13 in Varying Degrees of Osteoarthritis / 现代生物医学进展

Objective:To analysis the correlation of Wnt/β-catenin signaling pathway,OPN and MMP-13 in rabbit model with different degree of osteoarthritis (OA).Methods:Forty New Zealand white rabbits were randomly divided into four groups of 10 each,then we constructed the rabbit model with different degree of osteoarthritis (OA) by different concentrations of papain.The mRNA and protein levels of β-catenin,OPN,MMP-13,type Ⅱ collagen and proteoglycan were detected by Real-Time PCR and ELISA,respectively.Results:We have successfully established OA rabbit model by different concentrations of papain,and these OA rabbits model could be divided into mild,moderate and severe three level by Mankin scoring system,chondrocytes.Compared with the normal control group,the protein levels of β-catenin,OPN,and MMP-13 in the osteoarthritis were significantly increased,while the protein levels of type Ⅱ collagen and proteoglycan were significantly decreased (P＜0.05).With the increasing severity of OA,the results were consistent (P＜0.05).Conclusions:These results indicated that Wnt/β-catenin signaling pathway might regulate the expression of OPN,thereby affecting the expression of MMP-13,and ultimately have an impact on the occurrence ofosteoarthritis.

Total flavonoids from arachniodes exilispromotes osteogenic differentiation of hUCMSCs / 中国药理学通报

Aim To study the role of the total flavonoids from arachniodes exilis(TFAE) in osteogenic differentiation of human umbilical cord mesenchymal stem cells(hUCMSCs).Methods hUCMSCs were isolated and cultured by tissue explants adherent method, hUCMSCs at passage 3 were used to do the experiment, hUCMSCs were treated by different concentrations of TFAE, and cellular viability was measured by CCK 8 assay;alkaline phosphatase(ALP) activity was tested by AMP method;calcium nodule formation was detected by alizarin red staining;expression level of osteogenesis-related gene type I collagen enzyme a1(collagen type Ia1, Col1a1), osteopontin(OPN), Runx2, Osterix(Osx) mRNA was measured by RT-PCR;expression level of Col1a1,OPN protein was measured by Western blot.Results Certain concentration of TFAE(1 mg·L-1, 5 mg·L-1) promoted cellular proliferation, calcium nodules were more, ALP activity was enhanced;expression level of osteogenesis-related gene Col1a1, OPN, Runx2, OsxmRNA and Col1a1, OPN protein was up-regulated.Conculsion A certain concentration of TFAE promotes proliferation and osteogenic differentiation of hUCMSCs.

The relationships between the expression of osteopontin and esophageal squamous cell carcinoma and its clinicopathologic feature: A Meta-analysis / 中国医学装备

Objective:To systematic review the relationships between the expression of osteopontin (OPN) and esophageal squamous cell carcinoma and its clinicopathologic feature with meta-analyses.Methods: The published studies in the PubMed, the Cochrane Library, EMbase, CBM, CNKI, VIP and WanFang databases were searched, at the time, the documents retrospective method was adopted, and then a series researches about the relationships between the expression of OPN and esophageal squamous cell carcinoma and its clinicopathologic feature were collected. The Jan., 2017 was the deadline of search. The study arranged two researcher to screen documents as the standards of inclusion and exclusion, and extract data and evaluate the quality of methodology for these included documents. The RevMan 5.2 was applied to analyze these data in this Meta analysis.Results: A total of 12 case control studies were included, which included 782 cases of esophageal squamous cell carcinoma in case group and 700 persons in control group. The results of Meta analysis showed the OR of the expression of OPN between cases group and control group was 52.71, and its 95% confidence interval (CI) was 17.95-154.79. Besides, the OR of the expression of OPN between clinical Ⅰ-Ⅱ stage group and clinical Ⅲ-Ⅳ stage group was 0.20, and its 95% (CI) was 0.07-0.60. The OR of the expression of OPN between group with lymph node metastasis and group without lymph node metastasis was 6.37, and its CI was 3.64-11.15. The OR of the expression of OPN between T1+T2 group and T3+T4 group was 0.31, and its 95% (CI) was 0.15-0.64. The differences of the expression of OPN among various group had statistically significant.Conclusion:Current evidence indicates that the expression level of OPN is significantly correlated with esophageal squamous cell carcinoma and its clinicopathologic features. Due to the limited quantity of documents and quality of included studies, above conclusions need to be verified by conducting more high quality studies.

Expression of sodium iodide symporter mRNA and its clinical significance in differentiated thyroid carcinoma / 西安交通大学学报(医学版)

Objective To investigate the expression of sodium iodide symporter (NIS ) mRNA in differentiated thyroid Carcinoma (DTC)and further explore its value in clinical diagnosis and therapy of DTC. Methods The expression of NIS mRNA was detected and analyzed in 21 nodular goiter and 45 cases of DTC (including 35 cases of papillary thyroid carcinoma and 10 cases of follicular thyroid carcinoma)by using real-time fluorescent quantitative reverse transcription polymerase chain reaction (real-time RT-PCR).Results Compared with that in nodular goiter,the mRNA expression of NIS in DTC tissue was significantly decreased (P < 0.05 ). Moreover,the mRNA expression of NIS was significantly correlated with lymph node metastasis and AJCC stage, respectively.The expression of NIS mRNA in DTC with lymph node metastasis was significantly decreased compared with that in DTC without lymph node metastasis (P <0.05).In addition,the expression of NIS mRNA in Ⅱ-Ⅳstage DTC was significantly decreased compared with that in Ⅰ - Ⅱ stage DTC (P < 0.05 ).Conclusion Differential expression of NIS can provide evidence for individual 1 3 1 I therapy for DTC.

The expression level of OPN in serum of patients with oral squamous cell carcinoma / 实用口腔医学杂志

The concentration of osteopontin (OPN)in serum of patients with oral squamous cell carcinoma(n=58),benign maxillofacial tumor(n=45)and healthy controls(n=60)was examined using ELISA kit.The OPN level(ng/ml)in the 3 groups was 1 61 .8 ±1 2.6, 52.3 ±8.6 and 48.6 ±1 2.8(P<0.05)respectively,the cases with nodal metastasis and late clinical stage have relatively higher expression levels of OPN(P<0.05).Serum OPN may be related to OSCC progression.

Evaluation of the changes of osteopontin level after chemotherapy in patients with cervical cancer / 重庆医学

Objective To detect osteopontin (OPN) expression level of cervical cancer patients before and after chemotherapy , and explore the clinical value in the evaluation of chemotherapy efficacy .Methods 90 cases of cervical squamous cell carcinoma were treated with TP regimen ,immunohistochemical method was used to detect OPN expression before and after chemotherapy ,and studied the significance of OPN evaluation on curative effect in cancer chemotherapy .Results Before chemotherapy the positive ex-pression rate of OPN of tissue was 85 .56% ,and that after chemotherapy was 62 .22% ,the positive rate of OPN expression has sig-nificant difference before and after chemotherapy conclusion(χ2 =14 .118 ,P0 .05) .Patients with OPN negative expres-sion rate of CR was 100% (13/13) ,OPN patients with positive expression rate of CR was 45 .46% (35/77) ,compared two groups with statistical difference (P<0 .05) .Conclusion The expression of OPN in cervical cancer tissue in the tumor is related with chemotherapy sensitivity and effect of chemotherapy .

Expressions of OPN, COX-2 and CyclinD1 in breast infiltrating carcinoma and their relationship with clinicopathological features / 实用医学杂志

Objective To observe the expressions of OPN, COX-2 and CyclinD1 in breast infiltrating carcinoma and evaluate their relationships with clinic pathological features. Methods Expression of the above three indexes were detected from 70 breast cancinoma patients by immunohistochemistry. The relationships among them and clinicopathological features were analyzed. Results The positive expression rates of OPN were 78.8% in cases (≤45 years old) and 73.0% in cases (> 45 years old); the positive expression rates were 79.3%(tumor diameter ≤ 3 cm) and 73.2% (tumor diameter > 3 cm); the positive expression rates were 77.8%, 73.8% and 78.9% in cases ofⅠgrade, Ⅱgrade and Ⅲ respectively, the positive rates had no statistical significances(P > 0.05). The expression rates of OPN in cases of breast infiltrating carcinoma without and with axillary node metastasis were 62.5% and 93.3%, in cases at stage Ⅰ~Ⅱ and Ⅲ ~Ⅳ were, 68.0% and 95.0% respectively, the positive rates had statistical significances(P < 0.05). The expression of OPN was negatively correlated with ER and PR while positively correlated with CerbB2, COX-2 and CyclinD1. Conclusions OPN plays an important role in the invasion and metastasis of breast carcinoma coordinated with COX-2 and CyclinD1.

Osteopontin: A Novel Protein Molecule.

Osteopontin (OPN) is phosphorylated sialic acid –rich non-collagenous bone matrix protein. OPN is found in several biological fluids including human plasma, serum, breast milk and urine.OPN was named for its function as a bridge between cells and minerals. OPN has been implicated as an important factor in bone remodeling. Osteopontin is expressed in immune cells, including macrophages, neutrophils, dendritic cells with varying kinetics. OPN influences cell mediated immunity and has Th 1 cytokine functions .OPN is overexpressed in cancers of lung, breast, colorectal, stomach and ovary.OPN is found in atheromatous plaques within arteries. OPN plays an important role during both acute and chronic inflammation. OPN is upregulated in tissues during several pathological process including atherosclerosis, valve stenosis, myocardial infarction and rheumatic arthritis. OPN is a key cytokine regulating tissue repair. OPN’s plasma levels are elevated in overweight and obesity.

Different doses of vitamin K3 effect the expression of OPN of rats kidney / 中华泌尿外科杂志

Objective To investigate the effect of different doses of Vitamin K3 on the expression of osteopontin in rats kidney. Methods After different dose of Vitamin K3 were injected to different groups of rats feeded with same stone-inducing agent, the level of expression of OPN in renal tissues was observed with immunohistological staining. Results OPN expression located at distal convoluted tubule when the rats feeded without stone-inducing agent, and the OPN expression stain was extended to proximal convoluted tubule when the stone-inducing agent was feeded. Vitamin K3 can decrease the extension of OPN expression induced by the agent and more doses of Vitamin K3, more effects. Conclusions Vitamin K3 can decrease the expression of OPN in the kidney tubule of rats feeded with stone-induced agent.

Observing on relationship in changes of OPN,IL-18 levels with hepatic function in patients with chronic hepatic disease infected by HBV / 中国免疫学杂志

Objective:To observe clinical significance between changes of OPN,IL-18 levels and relation ship with hepatic function in patients with chronic hepatic disease infected by HBV.Methods:The levels of OPN,IL-18 in peripheral blood of 102 patients with chronic hepatitis B,34 patients with hepatitis B-related cirrhosis,95 HBV carriers and 20 healthy people as the control group were respectively detected with ELISA.Results:The levels of OPN and IL-18 in chronic hepatitis B and hepatitis B-related cirrhosis groups were significantly increased compared with that of health control group(P