Exosomes secreted from human umbilical cord mesenchymal stem cells promote pancreatic ductal adenocarcinoma growth by transferring miRNAs / 中华肿瘤杂志

Objective: To observe the effects of exosomes derived from human umbilical cord mesenchymal stem cells on the proliferation and invasion of pancreatic cancer cells, and to analyze the contents of exosomes and explore the mechanisms affecting pancreatic cancer cells. Methods: Exosomes extracted from human umbilical cord mesenchymal stem cells were added to pancreatic cancer cells BxPC3, Panc-1 and mouse models of pancreatic cancer, respectively. The proliferative activity and invasion abilities of BxPC3 and Panc-1 cells were measured by cell counting kit-8 (CCK-8) and Transwell assays. The expressions of miRNAs in exosomes were detected by high-throughput sequencing. GO and KEGG were used to analyze the related functions and the main metabolic pathways of target genes with high expressions of miRNAs. Results: The results of CCK-8 cell proliferation assay showed that the absorbance of BxPC3 and Panc-1 cells in the hucMSCs-exo group was significantly higher than that in the control group [(4.68±0.09) vs. (3.68±0.01), P<0.05; (5.20±0.20) vs. (3.45±0.17), P<0.05]. Transwell test results showed that the number of invasion cells of BxPC3 and Panc-1 in hucMSCs-exo group was significantly higher than that in the control group (129.40±6.02) vs. (89.40±4.39), P<0.05; (134.40±7.02) vs. (97.00±6.08), P<0.05. In vivo experimental results showed that the tumor volume and weight in the exosomes derived from human umbilical cord mesenchymal stem cells (hucMSCs-exo) group were significantly greater than that in the control group [(884.57±59.70) mm(3) vs. (695.09±57.81) mm(3), P<0.05; (0.94±0.21) g vs. (0.60±0.13) g, P<0.05]. High-throughput sequencing results showed that miR-148a-3p, miR-100-5p, miR-143-3p, miR-21-5p and miR-92a-3p were highly expressed. GO and KEGG analysis showed that the target genes of these miRNAs were mainly involved in the regulation of glucosaldehylation, and the main metabolic pathways were ascorbic acid and aldehyde acid metabolism, which were closely related to the development of pancreatic cancer. Conclusion: Exosomes derived from human umbilical cord mesenchymal stem cells can promote the growth of pancreatic cancer cells and the mechanism is related to miRNAs that are highly expressed in exosomes.

Scientific basis of acupuncture on mesenchymal stem cells for treating ischemic stroke / 中国针灸

The scientific basis of acupuncture on mesenchymal stem cells (MSCs) for treating ischemic stroke (IS) is discussed. MSCs transplantation has great potential for the treatment of tissue damage caused by early stage inflammatory cascade reactions of IS, but its actual transformation is limited by various factors. How to improve the homing efficiency of MSCs is the primary issue to enhance its efficacy. As such, the possible mechanisms of acupuncture and MSCs transplantation in inhibiting inflammatory cascade reactions induced by IS are explored by reviewing literature, and a hypothesis that acupuncture could promote the secretion of stromal cell-derived factor-1α (SDF-1α) from ischemic foci to regulate SDF-1α/CXC chemokine receptor 4 (CXCR4) axis, thereby improving the homing efficiency of MSCs transplantation, exerting its neuroprotective function, and improving the bed transformation ability, is proposed.

An experimental study on the biological characteristics of SDF-1/CS/β-GP composite bio-membrane in vitro / 口腔医学

Objective@#To prepare a composite membrane by chitosan/β-sodium glycerophosphate(CS/β-GP) thermosensitive hydrogel combined with stromal cell derived factor-1(SDF-1) and observe its biological characteristics in vitro.@*Methods@#Different doses of SDF-1 were added into CS/β-GP solution and then the thermosensitive gel time was measured. The SDF-1/CS/β-GP solution was membrane paved and dried to prepare composite membranes. The morphological characteristics were observed by scanning electron microscope(SEM). Composite membranes were placed into cell culture medium, and the supernatant(n=3) was extracted after standing at 6, 12, 24, 36, 48, 60 h, respectively. The concentration of SDF-1 in the solution was measured. Bone mesenchymal stem cells(BMSCs) were cultured in the Transwell room, and the composite membranes containing different concentrations of SDF-1 were placed in the lower chamber. There were four groups(n=3): Group M0 used CS/β-GP membrane(control group), Group M1, M2, M3 used SDF-1/CS/β-GP membrane(SDF-1 was 100, 200, 400 ng/mL respectively). After culture for 6, 12 and 24 h, the cells under the membrane were preserved and Giemsa stained and counted. The absorbance(OD) value was measured by MTT method to calculate the cell proliferation rate. SPSS 19.0 was used for multi-factor analysis of variance.@*Results @#After adding a certain amount of SDF-1 into CS/β-GP solution, the gel time did not change significantly(P>0.05). The SDF-1/CS/β-GP membrane was translucent and porous at 37 ℃. In this experiment, the volumic mass of SDF-1 released by SDF-1/CS/β-GP composite membrane increased gradually with the experimental time(P<0.01). Transwell cell chemotaxis test showed that the number of BMSCs cells with directional migration increased with the prolongation of observation time(P<0.01) and the increase of SDF-1 volumic mass(P<0.01). In MTT test, the OD value of migration cell solution increased with the prolongation of time(P<0.01) and the increase of SDF-1 volumic mass(P<0.01). @*Conclusion@# The SDF-1/CS/β-GP composite membrane has a porous structure and biological activity of chemotactic BMSCs directional migration. It is a potential membrane for guided tissue regeneration.

Effects of atractylodin on inflammatory damage of periodontal tissue and alveolar bone loss in periodontitis rats and its mechanism / 中国药房

OBJECTIVE To investigate the effects and mechanism of atractylodin on inflammatory injury of periodontal tissue and alveolar bone loss in periodontitis rats. METHODS A total of 144 SD rats were divided into control group （intragastric and intraperitoneal injection of normal saline）， model group （intragastric and intraperitoneal injection of normal saline）， atractylodin low-dose， medium-dose and high-dose groups （intraperitoneal injection of 6.665， 13.33， and 26.66 mg/kg atractylodin）， metronidazole group （positive control group， intragastric injection of 0.05 g/kg metronidazole， intraperitoneal injection of normal saline）， AMD3100 [stromal cell-derived factor-1 （SDF-1）/CXC chemokine receptor-4 （CXCR4） pathway inhibitor] group （intragastric injection of 1 mg/kg AMD3100， intraperitoneal injection of normal saline）， atractylodin high-dose+AMD 3100 group （intraperitoneal injection of 26.66 mg/kg atractylodin， intragastric injection of 1 mg/kg AMD3100）， with 18 rats in each group. Except for the control group， all other groups of rats were inoculated with Porphyromonas gingivalis to construct a periodontitis model. After successful modeling， they were given relevant medicine or normal saline， once a day， for 4 consecutive weeks. The gingival index of rats was detected； the levels of interleukin-6 （IL-6） and tumor necrosis factor α （TNF-α） in rat serum were also determined； alveolar bone resorption， periodontal histopathologic changes and the number of osteoclasts were detected by methylene blue staining， HE staining and TRAP staining， respectively. The expressions of osteoprotegerin （OPG）， receptor activator of NF-κB ligand （RANKL）， SDF-1 and CXCR4 proteins were determined. RESULTS Compared with the control group， serious pathological injury of periodontal tissue was found in the model group， the gingival index， the levels of IL-6 and TNF- α， alveolar bone absorption value， the number of osteoclasts， and the expression of RANKL protein were all increased significantly （P＜0.05）， while the expressions of OPG， SDF-1 and CXCR4 proteins were decreased significantly （P＜0.05）. Compared with the model group， pathological injury of periodontal tissue in rats was reduced； the gingival index， the levels of IL-6 and TNF-α， alveolar bone resorption value， osteoclast number and RANKL protein expression were decreased significantly， while protein expressions of OPG， SDF-1 and CXCR4 were increased significantly in atractylodin low-dose， medium-dose and high-dose groups and metronidazole group （P＜0.05）. The change trend of corresponding indexes in the AMD3100 group was opposite to the above （P＜0.05）. AMD3100 attenuated the inhibitory effect of high-dose atractylodin on inflammatory response and alveolar bone loss in rats with periodontitis （P＜0.05）. CONCLUSIONS Atractylodin may improve the inflammatory response and alveolar bone loss in periodontitis rats by activating the SDF-1/CXCR4 signaling pathway.

Risk factors of hemorrhagic cystitis after allo-HSCT and therapeutic effects of early transfusion of umbilical cord mesenchymal stem cells / 中华器官移植杂志

Objective:To explore the clinical efficacy and risk factors of umbilical cord mesenchymal stem cells (UCMSCs) infusion at an early stage (i.e.gross hematuria) for hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:The relevant clinical data were retrospectively reviewed for 300 patients undergoing allo-HSCT from January 2016 to July 2021.According to the presence or absence of HC, they were assigned into two groups of HC (n=89) and non-HC (control, n=211). According to whether or not receiving an infusion of UCMSCs, 51 patients of HC degree Ⅱ-Ⅳ were divided into two groups of UCMSC infusion and non-infusion.The risk factors of HC after allo-HSCT were analyzed by χ2 test.Logistic regression was employed for multivariate analysis of P<0.05.Mann-Whitney U test was utilized for statistically analyzing the duration of gross hematuria and urinary tract irritation symptoms and evaluating the clinical efficacy of UCMSCs infusion for HC. Results:Among them, 89 (29.67%) developed HC post-allo-HSCT.Clinical grades were Ⅰ (n=38, 42.70%), Ⅱ (n=36, 40.45%), Ⅲ (n=13, 14.61%) and Ⅳ (n=2, 2.25%). The median occurrence time was 29 (21.5-35.0) days post-allo-HSCT.In univariate analysis, age ≤30 years, haploid transplantation, antithymocyte globulin (ATG), acute graft-versus-host disease (aGVHD), CMV-DNA positive pretreatment significantly boosted the risk of HC ( P<0.05). In multivariate analysis, aGVHD was an independent risk factor for HC ( OR=10.281, 95% CI: 1.606-65.813, P=0.014). Among 89 HC patients, 38 grade Ⅰ patients were complete remission(CR). Among 51 patients of grade Ⅱ-Ⅳ HC, the outcomes were CR (n=48) and non-remission(NR)(n=3). And 24/51 of them received UCMSCs plus conventional treatment.The duration of gross hematuria was shorter in UCMSCs infusion group than that in UCMSCs non-infusion group [12(9-17) vs 17(12.0-26.5) day] and the difference was statistically significant ( P=0.045). And the duration of urinary tract irritation symptoms was shorter in UCMSCs infusion group than that in UCMSCs non-infusion group [18(11-30) vs 27(18.0-35.5) days] and the difference was statistically significant ( P=0.048). Conclusions:Indicated for post-ALLO-HSCT HC, infusion of UCMSCs may significantly shorten the course of disease.Age ≤30 years, haploid transplantation and preconditioning with positive ATG, aGVHD and CMV-DNA may boost the risks of HC post-allo-HSCT.And aGVHD is an independent risk factor for HC after allo-HSCT.

Comparative Evaluation of Mouse Bone Marrow Mesenchymal Stromal Cells Characteristics Cultured in Two Different Supplemented Media

@#Introduction: Preclinical studies on mesenchymal stromal cells (MSC) have allowed the cells to be considered as a promising candidate for cellular therapy. In recent years, conflicting data have been reported regarding various aspects of their characteristics, development and differentiation potential, which may be due to arrange of factors. Among the factors worth investigating is the culture medium formulation. Methods: Here we have made a comparative characterization of mouse bone marrow mesenchymal stromal cells (mBM-MSC) that were cultured using two common supplements, MesenCult™ Stimulatory Supplement (MSS) and fetal bovine serum (FBS), under the same experimental conditions at different passages. Clonogenic potential, cumulative population doubling level (CPDL), population doubling time (PDT), immunophenotyping, differentiation, immunosuppression potentials and chromosome analysis of early and late passages mBM-MSC were assessed. Results: Our findings showed that the CPDL, immunophenotype and immunosuppression potential of mBM-MSC were similar. However, variations were seen in their clonogenicity, population doubling time and differentiation efficacy whereby all of these were enhanced in DMSS. These observations suggest that their genetic make-up may be affected by both supplements upon prolonged culture. Interestingly, this conjecture was supported when chromosomal analysis revealed genetic instability of mBM-MSCs cultured in both supplements. Conclusion: In conclusion, culture medium formulation was shown to cause variations and spontaneous transformation in mBM-MSCs raising concerns on the usage of late passages mBMMSCs in fundamental and preclinical downstream experiments.

Effect of Acute Myeloid Leukemia Cells on the Proliferation and Apoptosis of Bone Marrow-Derived Mesenchymal Stromal Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of acute myeloid leukemia cells in leukemia-microenvironment on proliferation and apoptosis of bone marrow-derived mesenchymal stromal cells (BM-MSC).@*METHODS@#Acute myeloid leukemia (AML) murine models overexpressing MLL-AF9 were established. The number of BM-MSC of wild type (WT) and AML-derived mice were analyzed by flow cytometry. Morphology and growth differences between WT and AML-derived BM-MSC were analyzed by inverted fluorescence microscope. Proliferation and apoptosis of BM-MSC between these two groups were detected by Brdu and Annexin V/PI.@*RESULTS@#Compared with WT-derived BM-MSC, the number and proliferation rate of AML-derived BM-MSC significantly increased (P<0.01, P<0.001), while apoptosis rate decreased (P<0.05). When cultured in vitro, BM-MSC grew faster under conditional medium.@*CONCLUSION@#AML cells can promote proliferation and inhibit apoptosis of BM-MSC.

Research Advances on the Role of Bone Marrow Stromal Cell in Acute Lymphoblastic Leukemia --Review / 中国实验血液学杂志

Acute lymphoblastic leukemia (ALL) is a kind of the most common hematopoietic malignancy, its recurrence and drug resistance are closely related to the bone marrow microenvironment. Bone marrow stromal cell (BMSC) is an important part of the bone marrow microenvironment and their interaction with leukemia cells cannot be ignored. BMSC participates in and regulate signaling pathways related to proliferation or apoptosis of ALL cells by secretes cytokines or extracellular matrix proteins, thus affecting the survival of ALL cells. In this review, the research advance of several signaling pathways of the interaction between BMSC and ALL cells was summarized briefly.

Lowered expression of CCN5 in endometriotic tissues promotes proliferation, migration and invasion of endometrial stromal cells / 南方医科大学学报

OBJECTIVE@#To explore the expression of CCN5 in endometriotic tissues and its impact on proliferation, migration and invasion of human endometrial stromal cells (HESCs).@*METHODS@#We collected ovarian endometriosis samples from 20 women receiving laparoscopic surgery and eutopic endometrium samples from 15 women undergoing IVF-ET for comparison of CCN5 expression. Cultured HESCs were transfected with a recombinant adenovirus Ad-CCN5 for CCN5 overexpression or with a CCN5-specific siRNA for knocking down CCN5 expression, and the changes of cell proliferation, migration and invasion were evaluated using CCK-8 assay, wound healing assay and Transwell chamber assay. RT-qPCR and Western blotting were used to examine the expression levels of epithelial-mesenchymal transition (EMT) markers including E-cadherin, N-cadherin, Snail-1 and vimentin in HESCs with CCN5 overexpression or knockdown.@*RESULTS@#CCN5 expression was significantly decreased in ovarian endometriosis tissues as compared with eutopic endometrium samples (P < 0.01). CCN5 overexpression obviously inhibited the proliferation, migration and invasion of HESCs, significantly increased the expression of E-cadherin and decreased the expressions of N-cadherin, Snail-1 and vimentin (P < 0.01). CCN5 knockdown significantly enhanced the proliferation, migration and invasion of HESCs and produced opposite effects on the expressions of E-cadherin, N-cadherin, Snail-1 and vimentin (P < 0.01).@*CONCLUSION@#CCN5 can regulate the proliferation, migration and invasion of HESCs and thus plays an important role in EMT of HESCs, suggesting the potential of CCN5 as a therapeutic target for endometriosis.

Effects of psychotherapy combined with early comprehensive rehabilitation therapy on cerebral infarction / 中国基层医药

Objective:To investigate the clinical efficacy of psychotherapy combined with early comprehensive rehabilitation therapy in the treatment of acute cerebral infarction and the possible mechanism of action.Methods:Eighty-four patients with acute cerebral infarction who received treatment in Yuhang Fifth People's Hospital, China between June 2018 and February 2020 were included in this study. They were randomly divided into an observation group ( n = 44) and a control group ( n = 40). The control group was treated with conventional therapy. The observation group was subjected to early comprehensive rehabilitation therapy combined with psychotherapy based on conventional therapy. All patients were treated for 1 month. Clinical efficacy, the percentage of highly glycosylated type I transmembrane glycoprotein-positive cells and vascular endothelial growth factor receptor-2-positive cells (CD 34+KDR +) in monocytes, and serum level of stromal cell-derived factor-1α were compared between the observation and control groups. Results:After treatment, the scores of Hamilton Anxiety Scale (HAMA), Hamilton Depression Scale (HAMD) and National Institutes of Health Stroke Scale (NIHSS) were (16.4 ± 3.8) points, (17.9 ± 5.2) points, (3.56 ± 0.46) points, respectively, which were significantly lower than those in the control group [(23.4 ± 5.6) points, (23.7 ± 6.4) points, (5.39 ± 0.87) points, t = 7.896, 7.258, 6.935, all P < 0.05]. Barthel Index, Fugl-Meyer score, and Functional Independence Score in the observation group were (79.7 ± 20.8) points, (54.6 ± 17.2) points, (96.8 ± 8.5) points, respectively, which were significantly higher than those in the control group [(60.4 ± 17.6) points, (39.6 ± 14.8) points, (83.1 ± 9.7) points, t = 8.123, 7.251, 8.009, all P < 0.05]. After treatment, the percentage of CD34 +KDR + in monocytes and serum level of stromal cell-derived factor-1α in the observation group were (1.58 ± 0.19)% and (1.84 ± 0.11) μg/L, respectively, which were significantly higher than those in the control group [(0.73 ± 0.20)% and (1.34 ± 0.09) μg/L, t = 7.125, 6.983, both P < 0.05). Conclusion:Based on conventional treatment, psychotherapy combined with early rehabilitation treatment can improve the clinical efficacy in the treatment of acute cerebral infarction possibly through increasing the percentage of CD 34+KDR + in monocytes and the serum level of stromal cell-derived factor-1α.

Effect of Astragali Radix-Curcumae Rhizoma on SDF-1/CXCR4/NF-κB Signaling Pathway of Orthotopic Transplantation Model of Colon Cancer in Mice / 中国实验方剂学杂志

Objective:To explore the possible mechanism of Astragali Radix-Curcumae Rhizoma （AC） in inhibiting tumor growth in the orthotopic transplantation model of colon cancer in mice. Method:The molecular docking technology was used to predict the intermolecular interaction between the main active components of AC and the pathway target proteins， such as stromal cell-derived factor-1 （SDF-1）， C-X-C motif chemokine receptor 4 （CXCR4）， and nuclear factor kappa-B p65 （NF-<italic>κ</italic>B p65）. The orthotopic transplantation model of CT26.WT colon cancer was established in mice for <italic>in vivo</italic> experimental verification. Sixty BALB/c male mice were randomly divided into a sham operation group， a model group， a 5-fluorouracil （5-Fu， 30 mg·kg^-1） group，and low- （0.32 g·kg^-1）， medium- （0.64 g·kg^-1）， and high-dose （1.28 g·kg^-1） AC groups， with 10 mice in each group. The sham operation group and the model group received normal saline by gavage. The corresponding drugs were administered by gavage in the 5-Fu group and by intraperitoneal injection in the AC groups. After intervention for 15 days， the tumor <italic>in situ</italic> was completely stripped， and the colon tissues 5-6 cm in length adjacent to the tumor were taken. The tumor volume was measured and calculated. The pathological changes of tumor tissues and colon tissues were observed by Hematoxylin-Eosin （HE） staining. Western blot was used to detect the protein expression of SDF-1， CXCR4， p-NF-<italic>κ</italic>B p65 in colon tissues. Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） were used to detect SDF-1， CXCR4， NF-<italic>κ</italic>B p65， Cyclin D₁， oncogene c-Myc protein and mRNA expression in tumor tissues. Result:Compared with the model group， 5-Fu and AC groups showed reduced tumor volumes <italic>in situ</italic> （<italic>P</italic><0.05， <italic>P</italic><0.01）， with the tumor inhibition rate in the 5-Fu group as high as （61.38±2.34）%. The tumor-inhibiting effect was optimal in the medium-dose AC group， with the tumor inhibition rate of （43.43±3.71）%. Compared with the model group， 5-Fu and AC groups showed relieved pathological changes of tumor and colon tissues. Specifically， AC down-regulated the protein expression levels of SDF-1， CXCR4， and p-NF-<italic>κ</italic>B p65 in colon tissues （<italic>P</italic><0.01）， and down-regulated the protein and mRNA expression levels of SDF-1， CXCR4， NF-<italic>κ</italic>B p65， Cyclin D₁， and c-Myc in tumor tissues （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:AC can inhibit the growth of orthotopic transplantation tumor of colon cancer， and its intervention mechanism may be related to the regulation of related protein and mRNA expression in the SDF-1/CXCR4/NF-<italic>κ</italic>B signaling pathway.

Effect and Mechanism of Qixian Tongluo Prescription on Neural Function Recovery in Patients with Cerebral Infarction / 中国实验方剂学杂志

Objective:To investigate the effect of Qixian Tongluo prescription on neural function recovery in patients with cerebral infarction and its mechanism. Method:A total of 100 inpatients （January to June，2020）with cerebral infarction in the Neurology Department of Wenzhou Hospital of Traditional Chinese Medicine were assigned to an experimental group （<italic>n</italic>=50） and a control group （<italic>n</italic>=50） according to the random number table. Both groups received conventional treatment of western medicine，while the experimental group took additional Qixian Tongluo prescription. Treatment lasted for 12 weeks. The clinical efficacy，National Institutes of Health Stroke Scale （NIHSS） score， the modified Barthel index （MBI），Fugl-Meyer assessment （FMA） score， and levels of brain-derived neurotrophic factor（BDNF），vascular endothelial growth factor（VEGF）， and stromal cell-derived factor-1（SDF-1） in peripheral blood of the two groups before and after treatment were compared. Result:The total response rate in the experimental group was 84.00%（42/50），higher than 66.00%（33/50） in the control group （<italic>Z</italic>=-7.365，<italic>P</italic><0.05）. There was no significant difference in the scores of MBI，FMA， and NIHSS before treatment between the two groups. The MBI and FMA scores of the two groups increased （<italic>P</italic><0.01）， and the NIHSS scores decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the control group after treatment， the experimental group showed increased MBI and FMA scores and decreased NIHSS score （<italic>P</italic><0.05）. There was no significant difference in BDNF level between the two groups before and after treatment. The VEGF and SDF-1 levels in the peripheral blood of the two groups were higher than those before treatment （<italic>P</italic><0.05）， and the experimental group was higher than the control group （<italic>P</italic><0.05）. Conclusion:Qixian Tongluo prescription can effectively improve the clinical efficacy，the quality of life， and the prognosis of patients with cerebral infarction during convalescence. The underlying mechanism is associated with the promotion of the expression of endogenous VEGF and SDF-1 in the peripheral blood to activate the SDF-1/chemokine receptor 4（CXCR4） signaling pathway， induce the recruitment and mobilization of endothelial progenitor cells， and facilitate the angiogenesis and repair of ischemic brain tissues.

In vitro anti-tuberculosis effect of chitosan-gelatin/poly(lactic acid co-glycolic acid) combined with drug-loaded hydrogel / 中国组织工程研究

BACKGROUND: Anti-tuberculous chemotherapy is the main method for treating bone and joint tuberculosis. However, systemic administration hardly maintains the effective drug concentration in the focus area, and the therapeutic efficacy is unsatisfactory. OBJECTIVE: To prepare a chitosan-gelatin/poly(lactic-co-glycolic acid) combined with drug-loaded hydrogel, which can release anti-tuberculosis drugs in situ for a long time and promote osteogenesis. METHODS: Isoniazid, a hydrophilic anti-tuberculosis drug, and a hydrophobic stromal cell derived factor-1 were loaded into poly(lactic-co-glycolic acid) by double emulsion method to prepare drug-loaded poly(lactic acid co-glycolic acid) microspheres, which were then mixed into chitosan gelatin/poly(lactic acid co-glycolic acid) combined with drug-loaded hydrogel. The ability of drug delivery and anti-tuberculosis of poly(lactic acid co-glycolic acid) microspheres and chitosan gelatin/poly(lactic acid co-glycolic acid) combined with drug-loaded hydrogels in vitro were tested. MC3T3-E1 cells were inoculated on the surface of microspheres and hydrogel respectively. The biocompatibility was detected by cell counting kit-8 assay. The osteogenetic activity was detected by alkaline phosphatase activity. RESULTS AND CONCLUSION: (1) The burst release of isoniazid in the microspheres was about 23.3% in 1 hour, 42.6% in 2 days, and then it entered the sustained-release stage in the later 25 days. The burst release of stromal cell derived factor was about 19.8% in 1 hour, 44.7% in 2 days, and then it entered the sustained-release stage in the next 25 days. The release of isoniazid and stromal cell-derived factor in the combined drug-loaded hydrogel was 8.3% and 8.5% in the first hour, respectively. The cumulative release rates on the second day were 15.2% and 17.6%, respectively, which were much lower than that of poly(lactic acid co-glycolic acid) microspheres. (2) After 4 weeks in vitro, the antibacterial diameter of the combined drug-loaded hydrogel was much larger than that of the drug-loaded microspheres, and the antibacterial rate was higher than that of the drug-loaded microspheres (P < 0.05). (3) The combined drug-loaded hydrogel and the drug-loaded microspheres had good cytocompatibility and cell viability was about 100%. (4) After 5 and 10 days of culture, there was no significant difference in the activity of alkaline phosphatase on the surface of drug-loaded hydrogel and drug-loaded microspheres. (5) These results show that the in situ chitosan-gelatin/poly(lactic acid co-glycolic acid) combined with drug-loaded hydrogel can be used for treating tuberculosis and other bone and joint infections.

Expression and significance of stromal cell-derived factor-1 and its receptor CXCR4 in patients with pterygium / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the expression and significance of stromal cell-derived factor(SDF-1)and its receptor CXC chemokine receptor 4(CXCR4)in patients with pterygium of different ages.<p>METHODS: From January 2018 to October 2018, surgical specimens of 60 patients with primary pterygium and 60 eyes(including 30 eyes younger than 50 years old and 30 eyes older than 50 years old)were collected from the No.474 Hospital of Chinese PLA, at the same time collect age matched strabismus diorthosis and normal conjunctiva tissues of 30 patients with retinal detachment repairs(including 15 eyes younger than 50 years, older than 50 years of age 15 eyes).The expression and localization of SDF-1/CXCR4 in pterygium tissue specimens were detected by HE staining and immunohistochemistry, and the relationship between the expression of SDF-1/CXCR4 and the clinical characteristics of patients was analyzed. The mean optical density of SDF-1 /CXCR4 was measured by IPP 6.0 software.<p>RESULTS: SDF-1/CXCR4 showed slightly positive or no positive expression in normal conjunctival basal cells, but positive expression in both full-layer conjunctival epithelial cells and vascular endothelial cells in pterygium, with significant difference in expression level, and more obvious expression in basal cells, showing obvious polarity. The expression levels of SDF-1 and CXCR4 in pterygium tissues were higher than those in normal conjunctival tissues, and the difference was statistically significant(<i>P</i><0.05). The expression of CXCR4 in patients younger than 50 years old was greater than that in patients older than 50 years old, and the difference was statistically significant(<i>P</i><0.05).<p>CONCLUSION: SDF-1 and CXCR4 expression is up-regulated in pterygium, suggesting that SDF-1 and CXCR4 participates in the formation of pterygium and inhibits the SDF-1/CXCR4 signaling pathway, which may inhibit the occurrence of pterygium, and may also become a drug therapeutic target for pterygium, and become a new research direction. The higher expression of CXCR4 in young pterygium patients suggests that individualized drug administration may be realized in the future to reduce the waste of medical resources.

Effect of Duhuo Xuduan Tang for Oral Administration and Iontophoresis on SDF-1/CXCR4 Signaling Pathway and Relevant Factors of Cartilage Degradation in Knee Joint Fluid of Patients with Knee Osteoarthritis with Liver and Kidney Deficiency / 中国实验方剂学杂志

Objective::To observe the clinical efficacy of Duhuo Xuduan Tang for oral administration and iontophoresis in the treatment of knee osteoarthritis (KOA) with liver and kidney deficiency and its effect on stromal cell-derived factor-1 (SDF-1)/C-X-C chemokine receptor type 4 (CXCR4) signaling pathway. Method::Totally 150 KOA patients with deficiency of liver and kidney diagnosed in the Teaching Hospital of Tianjin University of Traditional Chinese Medicine(TCM) were randomly divided into control group, oral TCM group and iontophoresis group, with 50 cases in each group. The control group was given glucosamine sulfate capsule, 0.5 g/time, twice a day, while the oral TCM group was given Duhuo Xuduan Tang, 150 mL/time, twice a day. In the iontophoresis group, Duhuo Xuduan Tang was administered at Kuangu acupoint, Xiguan acupoint, Xiyan acupoint and Dubi acupoint for iontophoresis for 30 minutes, once a day. All of the three groups were treated for 4 weeks. The swelling degree and the pain degree of knee joint before and after treatment were observed, and the clinical efficacy was recorded. The protein contents of SDF-1, CXCR4, matrix metalloproteinase-3 (MMP-3) and matrix metalloproteinase-13 (MMP-13) in knee joint fluid before and after treatment were detected by enzyme-linked immunosorbent assay (ELISA). Result::The efficacy of oral TCM group was better than that of iontophoresis group and control group, and the recurrence rate was the lowest (P<0.05). Compared with before treatment, the tenderness increased, whereas visual analogue scale(VAS) score, knee swelling score, The Western Ontario and McMaste Universities (WOMAC) score and SDF-1, CXCR4, MMP-3 and MMP-13 protein content in knee joint fluid decreased in oral TCM group after treatment, which were better than those in iontophoresis group and control group (P<0.05). Conclusion::Duhuo Xuduan Tang for oral administration and iontophoresis has an obvious effect on KOA with liver and kidney deficiency, with the best effect through oral administration. Its mechanism may be related to the inhibition of SDF-1/CXCR4 inflammatory signaling pathway and cartilage decomposition.

Effect of Ru′ai Shuhou Prescription on Proliferation and Metastasis of Breast Cancer MDA-MB-453 Cells Through SDF-1/CXCR4 Biological Axis / 中国实验方剂学杂志

Objective:To investigate the effect of Ru′ai Shuhou prescription （RSR） drug-containing serum on the proliferation and invasion ability of breast cancer cells MDA-MB-453 based on the biological axis of stromal cell-derived factor-1（SDF-1）/chemokine receptor 4 （CXCR4）. Method:A model of MDA-MB-453 cells with SDF-1-induced high expression of CXCR4 was established， and the rat drug-serum containing RSR and blank rat serum were prepared respectively. The cells were divided into fetal bovine serum control group （Blank）， blank rat serum group， SDF-1+blank rat serum group， SDF-1+RSR group， AMD3100+ SDF-1+blank rat serum group， and AMD3100+ SDF-1+RSR group. After intervention for 48 h， cell proliferation was detected by cell counting kit-8 （CCK-8） assay， cell invasion ability was detected by transwell assay， and mRNA and protein expressions of CXCR4， matrix metalloproteinase-2 （MMP-2） and MMP-9 were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. Result:As compared with the blank serum group， the proliferation of MDA-MB-453 cells was promoted and expression of CXCR4 mRNA was increased significantly when SDF-1 was 100 μg·L-1 （P<0.05）. As compared with SDF-1+blank rat serum group， RSR inhibited the proliferation and invasion of MDA-MB-453 cells induced by SDF-1， and at the same time， down-regulated the mRNA and protein expressions of CXCR4， MMP-2 and MMP-9 （P<0.05）. After pre-treatment with AMD3100 for 24 h， the inhibitory effect of RSR to cell proliferation was significantly increased （P<0.05）， and meanwhile， the decreases in mRNA and protein expression of CXCR4， MMP-2 and MMP-9 were more obvious， with statistically significant differences （P<0.05）. Conclusion:Through SDF-1/CXCR4 biological axis， RSR could down-regulate the expression of MMP-2 and MMP-9， reduce the degradation of extracellular matrix （ECM）， and then inhibit the metastasis of MDA-MB-453 cells. In addition， it has a synergistic effect with CXCR4 inhibitor AMD3100.

In vitro study on promoting migration ability of rat adipose derived stem cells modified by stromal cell-derived factor 1α / 中国修复重建外科杂志

Objective: To explored the effect of stromal cell-derived factor 1α (SDF-1α) on promoting the migration ability of rat adipose derived stem cells (rADSCs) by constructed the rADSCs overexpression SDF-1α via adenovirus transfection. Methods: rADSCs were isolated from adipose tissue of 6-week-old SPF Sprague Dawley rats. Morphological observation, multi-directional differentiations (osteogenic, adipogenic, and chondrogenic inductions), and flow cytometry identification were performed. Transwell cell migration experiment was used to observe and screen the optimal concentration of exogenous SDF-1α to optimize the migration ability of rADSCs; the optimal multiplicity of infection (MOI) of rADSCs was screened by observing the cell status and fluorescence expression after transfection. Then the third generation of rADSCs were divided into 4 groups: group A was pure rADSCs; group B was rADSCs co-cultured with SDF-1α at the best concentration; group C was rADSCs infected with recombinant adenovirus-mediated green fluorescent protein (Adv-GFP) with the best MOI; group D was rADSCs infected with Adv-GFP-SDF-1α overexpression adenovirus with the best MOI. Cell counting kit 8 (CCK-8) and Transwell cell migration experiment were preformed to detect and compare the effect of exogenous SDF-1α and SDF-1α overexpression on the proliferation and migration ability of rADSCs. Results: The cell morphology, multi-directional differentiations, and flow cytometry identification showed that the cultured cells were rADSCs. After screening, the optimal stimulating concentration of exogenous SDF-1α was 12.5 nmol/L; the optimal MOI of Adv-GFP adenovirus was 200; the optimal MOI of Adv-GFP-SDF-1α overexpression adenovirus was 400. CCK-8 method and Transwell cell migration experiment showed that compared with groups A and C, groups B and D could significantly improve the proliferation and migration of rADSCs ( P<0.05); the effect of group D on enhancing the migration of rADSCs was weaker than that of group B, but the effect of promoting the proliferation of rADSCs was stronger than that of group D ( P<0.05). Conclusion: SDF-1α overexpression modification on rADSCs can significantly promote the proliferation and migration ability, which may be a potential method to optimize the application of ADSCs in tissue regeneration and wound repair.

Effect of ranibizumab combined with vitrectomy on the serum VEGF-A and SDF-1 expression in patients with proliferative diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To discuss the effect of ranibizumab combined with vitrectomy(VT)on the serum vascular endothelial growth factor A(VEGF-A)and human stromal cell derived factor 1(SDF-1)expression in patients with PDR.<p>METHODS: Totally 120 patients with PDR were selected from January 2017 to January 2018 in our hospital, according to the random digital table, they were divided into group A and group B, 60 cases in each group, group A was given VT treatment, group B was given ranibizumab vitreous injection treatment on this basis, the VT operation, complication and VEGF-A, SDF-1, BCVA, CMT were compared between the two groups.<p>RESULTS: The postoperative serum VEGF-A and SDF-1 levels in the two groups were significantly lower than those in the preoperative, and the group B was significantly lower than group A(<i>P</i><0.05). The VT operation time, electrocoagulation, intraoperative bleeding and complication rate in the group B were significantly lower than those in the group A(<i>P</i><0.05). The 1d, 3mo postoperative BCVA, CMT in the two groups were significantly lower than those in the preoperative, and the group B was significantly lower than the group A(<i>P</i><0.05).<p>CONCLUSION: ranibizumab combined with VT can effectively improve the serum VEGF-A and SDF-1 levels in patients with PDR, it can reduce the trauma and complications of VT, and improve the BCVA and CMT in patient.

Activation of farnesoid X receptor by GW4064 inhibits invasive growth of colon cancer cells and expression of stromal cell-derived factor 1 / 第二军医大学学报

Objective To explore the mechanism by which specific agonist of farnesoid X receptor (FXR) GW4064 inhibits the growth and invasion of colon cancer cells. Methods Human colon cancer cell lines HT-29 were in vitro cultured. After treatment with GW4064 of 0, 0.1, 1, 3, 5, 7 and 10 μmol/L for 72 h, cell viability was measured by MTT assay. After treatment with GW4064 of 0, 1 and 5 μmol/L for 24 h, the cell morphology was observed under phase contrast microscope, and the mRNA expression levels of FXR and stromal cell-derived factor 1 (SDF-1) were detected by PCR. After treatment with GW4064 of 0 and 1 μmol/L for 24 h, the cell invasive ability was detected by cell scratch test. After treatment with GW4064 of 0, 1, 5 and 7 μmol/L for 24 h, the SDF-1 expression in the culture medium was detected by ELISA. Nude mouse. tumorigenesis model was established by subcutaneous inoculation of HT-29 cells. After intragastric administration with GW4064 or DMSO for 16 d, the tumor growth and the mRNA expression of FXR and SDF-1 in the tumors were determined. ResultsGW4064 inhibited the growth of HT-29 cells in a dose-dependent manner, and there was significant difference in the cell viability of HT-29 cells between the GW4064 groups (1, 3, 5, 7 and 10 μmol/L) and the control group (0 μmol/L, all P<0.05). After treatment with GW4064, phase contrast microscopy showed contracted and rounded colon cancer cells and slender cells transforming into epidermoid cells. The cell scratch test showed that the invasion ability of the colon cancer cells was significantly reduced after treatment with GW4064 compared with the control group (0 μmol/L, P<0.05). PCR results showed that the mRNA expression level of FXR was increased in a dose-dependent manner after GW4064 treatment, while the expression of SDF-1 mRNA changed in the opposite way. ELISA results showed that the SDF-1 expression in the cell culture supernant was decreased with the increase of GW4064 concentrations, and there were significant differences between the GW4064 (1, 5 and 7 μmol/L) groups and the control group (0 μmol/L, P<0.05). GW4064 significantly reduced tumor size compared with the control group (DMSO, P<0.05). In addition, the mRNA expression of FXR in the tumors was increased, and the mRNA expression of SDF-1 was decreased. ConclusionThe activation of FXR can inhibit the invasive growth of colon cancer cells and the expression of SDF-1.