BACKGROUND:Small diameter artificial vessels are urgently needed to treat coronary artery and peripheral artery diseases in clinical practice.At present,vascular tissue engineering has become the main method for preparing small diameter artificial vessels.Selecting suitable biomaterials and cell sources is the key factor for successful construction of small diameter tissue engineered vessels. OBJECTIVE:To observe the effect of four kinds of electrospinning membrane materials on proliferation,adhesion and differentiation of bone marrow mesenchymal stem cells into vascular smooth muscle cells. METHODS:Bone marrow mesenchymal stem cells were isolated and extracted from SD rats.The bone marrow mesenchymal stem cells were inoculated separately on polycaprolactone(PCL),polycaprolactone-hyaluronic acid(PCL-HA),polycaprolactone-silk-filament proteins(PCL-SF),and polycaprolactone-gelatin(PCL-GEL)electrospinning membrane materials.After 1,3,and 7 days of culture,the cell arrangement on the material was observed under scanning electron microscope.The proliferation and adhesion of the material were observed by phalloidin staining.The mRNA expressions of CD90,Meflin,and transforming growth factor β were detected by qRT-PCR.After 7 days of induced differentiation into vascular smooth muscle cells,the mRNA expression ofɑ-smooth muscle actin on the material was detected by qRT-PCR. RESULTS AND CONCLUSION:(1)Bone marrow mesenchymal stem cells were arranged along the fibers of the four kinds of electrospinning membranes under scanning electron microscopy.(2)Phalloidin staining showed the regular distribution of bone marrow mesenchymal stem cells on the four kinds of electrospinning membranes and parallel distribution along the fiber direction.Moreover,PCL-HA,PCL-SF,and PCL-GEL electrospinning membranes were more conducive to the proliferation and adhesion of bone marrow mesenchymal stem cells than PCL electrospinning membranes.Compared with PCL-HA and PCL-GEL electrospinning membranes,PCL-SF electrospinning membranes were more conducive to the proliferation and adhesion of bone marrow mesenchymal stem cells.(3)qRT-PCR showed that the four kinds of electrospun membrane materials could maintain the mRNA expression of CD90 and Meflin in bone marrow mesenchymal stem cells,but there was no significant difference between groups(P>0.05).The mRNA expression of transforming growth factor β in PCL-HA,PCL-SF,and PCL-GEL groups was higher than that in PCL group on days 1 and 7(P<0.05),and the mRNA expression of transforming growth factor β in PCL-SF group was higher than that in the other three groups on days 3 and 7(P<0.05).The mRNA expression of transforming growth factor β in PCL-HA group was higher than that in PCL-GEL group on day 7(P<0.05).(4)qRT-PCR showed that the mRNA expression of ɑ-smooth muscle actin in PCL-SF group was higher than that in the other three groups(P<0.05),and that in PCL-HA group was higher than that in PCL group(P<0.05).(5)The results indicate that compared with PCL,PCL-HA and PCL-GEL electrospinning membranes,PCL-SF electrospinning membranes combined with bone marrow mesenchymal stem cells are more suitable for the preparation of small diameter tissue engineered vessels.

As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate?adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.

Objective:To investigate the effect of high-intensity interval exercise(HIIT)on necroptosis and neuroinflam-mation in hippocampus of type 2 diabetes mellitus(T2DM)mice. Method:Male C57BL/6 mice fed with high-fat for ten weeks and then injected with streptozotocin(STZ)to establish the model of T2DM mice.Mice were divided into normal control group(CON,n=10),T2DM mod-el group(T2DM,n=10),and T2DM exercise group(HIIT,n=10).Mice in HIIT group received treadmill training for 7 weeks.At the end of the experiment,the necroptosis of hippocampal neurons was determined by lactate dehydrogenase(LDH)content and propidium iodide(PI)staining.The NOD-like receptor protein 3(NLRP3)level was examined by immunofluorescence staining.The mRNA expressions of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in hippocampus were evaluate by quantitative real-time polymerase chain reaction(qRT-PCR).The level of necroptosis and inflammatory cytokines related proteins in hippocam-pus were measured by the Western Blot. Result:Compared with the CON group,the LDH release level was significantly increased(P＜0.0001),the number of PI-stained positive cells and the expression level of necroptosis related proteins were increased in T2DM group.The fluorescence intensity of NLRP3,the expression of NLRP3,caspase-1,cleaved caspase-1 protein and inflammatory cytokines iNOS,IL-6,TNF-α and IL-1β were significantly increased(P＜0.05)in T2DM group.After 7 weeks of the HIIT intervention,the hippocampal LDH release level of HIIT group was significantly lower than T2DM group(P＜0.0001),the number of PI-stained positive cells and the expressions level of necroptosis related proteins was also decreased.The fluorescence intensity of NLRP3 and the expres-sion of NLRP3,caspase-1,Cleaved caspase-1 protein and the expression levels of proinflammatory factor were all decreased in HIIT group compared to the T2DM group(P＜0.05). Conclusion:Necroptosis was found in the hippocampal tissue of T2DM mice.HIIT can inhibit the necroptosis and eventually reduce the inflammatory response in the hippocampal tissue of T2DM mice.

Platycodon grandiflorum (Jacq.) A. DC. is a famous medicinal plant commonly used in East Asia. Triterpene saponins isolated from P. grandiflorum are the main biologically active compounds, among which polygalacin D (PGD) has been reported to be an anti-tumor agent. However, its anti-tumor mechanism against hepatocellular carcinoma is unknown. This study aimed to explore the inhibitory effect of PGD in hepatocellular carcinoma cells and related mechanisms of action. We found that PGD exerted significant inhibitory effect on hepatocellular carcinoma cells through apoptosis and autophagy. Analysis of the expression of apoptosis-related proteins and autophagy-related proteins revealed that this phenomenon was attributed to the mitochondrial apoptosis and mitophagy pathways. Subsequently, using specific inhibitors, we found that apoptosis and autophagy had mutually reinforcing effects. In addition, further analysis of autophagy showed that PGD induced mitophagy by increasing BCL2 interacting protein 3 like (BNIP3L) levels.In vivo experiments demonstrated that PGD significantly inhibited tumor growth and increased the levels of apoptosis and autophagy in tumors. Overall, our findings showed that PGD induced cell death of hepatocellular carcinoma cells primarily through mitochondrial apoptosis and mitophagy pathways. Therefore, PGD can be used as an apoptosis and autophagy agonist in the research and development of antitumor agents.
Humans ; Mitophagy ; Carcinoma, Hepatocellular/pathology* ; Liver Neoplasms/pathology* ; Cell Line ; Autophagy ; Apoptosis ; Membrane Proteins ; Proto-Oncogene Proteins/genetics* ; Tumor Suppressor Proteins/pharmacology*

Objective:To investigate the expression and its diagnostic value of long-chain non coding RNA (lncRNA) and colon cancer-related transcript-2 (CCAT2) in serum of patients with cervical cancer (CC).Methods:Serum samples of 100 CC patients, 60 CIN patients and 80 healthy people enrolled by Nantong Tumor Hospital from January 2016 to June 2017 were collected.The expression levels of CCAT2 in sera of CC patients and their corresponding postoperative patients, CIN patients and healthy controls were detected by real-time fluorescent quantitative PCR (RT-qPCR). The correlation between CCAT2 and clinicopathological features, as well as the traditional auxiliary diagnostic makers of CC, such as carbohydrate antigen 125 (CA125) and squamous cell carcinoma antigen (SCC) was analyzed. The working characteristic curve (ROC) of the subjects was used to evaluate the CCAT2 pair in the auxiliary diagnostic value of cervical cancer.Results:The relative expression levels of serum CCAT2 in patients with cervical cancer, patients after operation, patients with CIN and healthy controls were1.689 (0.616, 4.776), 1.018 (0.227, 3.328), 0.815 (0.453, 1.266) and 0.740 (0.271, 1.670), respectively.The relative expression of CCAT2 in cervical cancer patients was significantly higher than that in post-operative patients, CIN patients and healthy controls. The difference was statistically significant( t=6.999,8.193,9.345, P<0.001).There was no significant difference in the relative expression of CCAT2 between CIN patients and healthy controls ( t=0.327, P>0.05).The relative expression of CCAT2 in serum of cervical cancer patients had no significant difference in age 1.636(1.000,2.370),1.705(1.095,2.243) (χ 2=0.137, P=0.712) and menopause 1.672(1.059,2.342),1.659(1.068,2.298) (χ 2=0.000, P=1.000), but had significant difference with tumor size expression1.189(0.916,1.725),2.019(1.537,2.497)(χ 2=17.508, P=0.000),International Federation of Obstetrics and Gynecology (FIGO) staged expression stage 0.993(0.779,1.266),2.056(1.547,2.549),3.987(3.699,4.275)(χ 2=36.075, P=0.000) and lymph node metastasis 1.434(1.007,2.251),2.019(1.731,3.098) (χ 2=8.634, P=0.003). There was no correlation between the relative expression of serum CCAT2 and CA125 ( r2=0.003, P=0.563) and SCC (r 2=0.128, P=0.000).The diagnostic efficacy of serum CCAT2, CA125 and SCC in cervical cancer patients was analyzed by ROC curve. When compared with CIN patients, the areas under the curve were 0.890, 0.549 and 0.744, respectively. When compared with healthy patients, the areas under the curve were 0.857, 0.650 and 0.758, respectively. Conclusions:The level of serum CCAT2 in patients with cervical cancer is significantly higher than that in patients with cervical cancer, CIN patients and healthy controls. Serum CCAT2 may be a relevant marker for the diagnosis and prognosis of cervical cancer.

Objective: To investigate the association of periodontal diseases in pregnant women with preterm birth and low birth weight. Methods: 213 pregnant women aged 22-40 years were enrolled while receiving periodontal examination by 26 weeks of gestation. Periodontal measurements included plaque index(PLI),bleeding on probing (BOP),probing pocket depth(PD) and clinical attachment loss(CAL) were used as the criteria to classify the groups: healthy group(HG; n = 63),gingivitis group(GG; n = 74) and periodontitis group(PG; n = 76). At delivery,birth date and birth weight were recorded. Results: Preterm birth ratio in PG group is higher than that in HG group and GG group(P ＜ 0. 05). Conclusion: Periodontal disease may lead to preterm birth in pregnant women.

Exosomes are small (30 -100 nm) membrane vesicles secreted by various cell types , they mediate cell-to-cell communication by transferring mRNAs , miRNAs, and proteins.As a hotspot in the research of oncology , exosomes have potential values in the research of development , diagnosis and treatment of cancer. This paper briefly reviews the relationship between breast cancer microenvironment , drug resistance and exosomes and its progress in breast cancer diagnosis and treatment , in order to propose new diagnostic and therapeutic strategies .

Objective To explore the application effect of PDCA cycles on the normal limb position of stroke patients with hemiplegia. Methods The stroke patients with hemiplegia (128 cases) were selected as study subjects. Patients(62 cases)during January to September 2014 were set as the control group,and received routine nursing care. Patients(66 cases)during October 2014 to July 2015 were set as the experimental group,and used PDCA cycles management on the normal limb position additionally. The application effect of PDCA cycles on the normal limb position was evaluated through comparing two groups with qualification rates of normal limb position and incidence of complications. Results The qualification rates of normal limb position in the control group was 38.71%(24/62), which was higher than that of the control group, which was 75.76% (50/66) (χ2=16.504, P<0.01). The complications occurred in the control group were strephenopodia (11 cases), foot drop (16 cases), dislocation of shoulder (9 cases), omodynia (27 cases) and myospasm (34 cases), and they were 3 cases, 7 cases, 2 cases, 15 cases and 18 cases in the experimental group respectively. The incidence of complications was lower than those of the control group (χ2=4.001-8.961, P < 0.05), and the difference was statistically significant between two groups. Conclusions PDCA cycles management could improve the qualification rates of normal limb position and reduce the incidence of complications, which was beneficial to the recovery of limb function.

Objective To explore the expression level of serum miR135a?5p and its diagnostic value in colorectal cancer ( CRC) . Methods Serum samples were randomly collected from 60 primary CRC patients, 40 patients with intestinal polyps and 50 healthy controls, and the serum concentrations of miR135a?5p, CEA and CA199 were detected. The relationships between serum miR135a?5p level and clinicopathological parameters were analyzed by Mann?Whitney U test. The correlation of serum miR135a?5p level and serum concentrations of CEA or CA199 was analyzed by Pearson ’ s correlation test. Receiver operating characteristic ( ROC) curves and area under the curve ( AUC) were used to evaluate the diagnostic efficacy of miR135a?5p, CEA and CA199 as diagnostic indicators. Results The serum level of miR135a?5p in CRC patient was 2.451 (1.107, 4.413), significantly higher than 0.946 (0.401, 1.942) in the patients with intestinal polyps and 0. 949 ( 0. 194, 1. 415) in the healthy controls ( U = 351. 0 and U = 313. 0, respectively, P<0. 001 ) . The serum level of miR135a?5p in CRC patients was associated with both histological differentiation and clinical stage (P<0.05 for both), however, not correlated with the serum concentration of CEA (r2= 0.023, P = 0.293) or CA199 (r2= 0.067,P = 0.068). The AUC of serum miR135a?5p level in CRC patients was 0.832 (0.730-0.930) when compared to the patients with intestinal polyps and was 0.875 (0.800-0.950) when compared with the healthy controls. Conclusions The serum level of miR135a?5p in CRC patients is significantly higher than that in patients with intestinal polyps and healthy controls, and might be an important diagnostic marker of CRC.

Objective To explore the role of miR-202 in multiple myeloma (MM) cells,and study the regulation of miR-202 on drug sensitivity of MM cells.Methods miR-202 and BAFF mRNA levels were detected by real-time PCR.U266 cells were transfected with miR-202-mimics,miR-202-inhibitor,siBAFF and their negative controls.After above treatments,protein levels of Bcl-2 family and MAPK signaling pathway were detected by Western blot analysis,and the proliferation and apoptosis ability of MM cells were examined by WST-1,Annexin Ⅴ-FLUOS assay,respectively.Results The results showed that the expression ofmiR-202 in CD138+ MM cells (0.304±0.354) and U266 cells (0.052± 0.009) were lower than in normal controls (3.550± 1.126) (P＜0.001,P=0.009),whereas BAFF mRNA levels (5.700±0.734,9.576±2.887) were higher than in normal controls (1.819±0.853) (P＜0.001,P=0.006).The proliferation ability of U266 cells transfected with miR-202 mimics was significantly inhibited than in control group [(56.04±0.021)％ vs (18.89±0.32)％,P=0.002].The result of Western blot showed that the expression of Bcl-2 decreased by about 24％,and the expression of Bax increased by about 124％ in cells transfected with miR-202 mimics.The apoptosis rate in cells transfected with miR-202 mimics was significantly more than in control group [(49.60±4.89)％ vs (26.20±1.28)％,P=0.029].The apoptosis rate in miR-202 mimics combined with Bort group (51.23±5.41)％ was higher as compared with Bort treatment alone (31.70±4.40) ％ or miR-202 mimics control combined with Bort group (27.94±4.04) ％,(P=0.047,P=0.028),whereas the apoptosis rate in miR-202 mimics combined with Thal or Dex had no significant difference compared with miR-202 mimics control [(11.66±1.91)％ vs (10.63±1.74)％,P=0.700;(16.35± 1.32)％ vs (17.43 ± 1.95)％,P=0.400].The inhibitory rate of cell growth in miR-202 mimics combined with Bort group was higher as compared with Bort treatment alone [(36.93±5.98)％ vs (18.18±4.10)％,P=0.029].The expressions of p-JNK protein decreased in U266 cells transfected with miR-202 mimics and treated with Bort.Conclusion miR-202 mimics combined with Bort could inhibit proliferation and induce apoptosis of U266 cells through negative regulating target gene BAFF,which further inhibited the JNK/ SAPK signaling pathway.