MiRNA-128-3p induces osteogenic differentiation of bone marrow mesenchymal stem cells via activating the Wnt3a signaling.

OBJECTIVE: To clarify the biological function of miRNA-128-3p in influencing the progression of osteoporosis by inducing osteogenic differentiation of MSCs via activating the Wnt3a signaling. PATIENTS AND METHODS: Dynamic expression levels of miRNA-128-3p in osteogenically differentiated MSCs at the different time points were detected by qRT-PCR. The binding sites in the seed sequence of miRNA-128-3p and Wnt3a were predicted using the bioinformatic tool, and their interaction was further confirmed by Dual-Luciferase reporter assay. Co-regulation of miRNA-128-3p and Wnt3a on relative levels of osteogenesis-associated genes, ALP activity and mineralization ability in glucocorticoid-induced MSCs were assessed. RESULTS: MiRNA-128-3p was gradually upregulated with the prolongation of osteogenic differentiation of MSCs. Overexpression of miRNA-128-3p reversed the declines in glucocorticoid-induced expression levels of osteogenesis-associated genes (Bglap, RUNX2 and BMP-2), ALP activity and mineralization ability in MSCs. Wnt3a was able to bind miRNA-128-3p. Its level was positively regulated by miRNA-128-3p in MSCs. Enhanced ALP activity and mineralization ability in glucocorticoid-induced MSCs overexpressing Wnt3a were partially abolished by knockdown of miRNA-128-3p. CONCLUSIONS: By positively regulating Wnt3a, miRNA-128-3p alleviates the progression of osteoporosis through inducing osteogenic differentiation of MSCs.

Association between C-myc and K-ras gene polymorphisms and non-Hodgkin lymphoma.

OBJECTIVE: To explore the association between c-myc and K-ras gene polymorphisms and non-Hodgkin lymphoma (NHL). PATIENTS AND METHODS: A total of 200 NHL patients in our hospital in the past 3 years were collected as disease group, while 200 healthy people were taken as control group. The genomic deoxyribonucleic acid (DNA) in the peripheral blood was extracted in both groups, amplified via Polymerase Chain Reaction (PCR) and sent to the company for the detection of c-myc and K-ras gene polymorphisms. The expressions of c-myc and K-ras were detected via Reverse Transcription-quantitative PCR (RT-qPCR), and the levels of clinical indexes hemoglobin (Hb), platelet (PLT) and lactate dehydrogenase (LDH) were determined in the Laboratory Department. RESULTS: The allele distribution at c-myc gene locus rs121918684 was different between control group and disease group (p=0.000), and the G allele frequency was 202 (0.505) in the control group and 263 (0.657) in the disease group. In the disease group, the GG genotype frequency at c-myc gene locus rs121918684 [97 (0.485)], the CC genotype frequency at rs775522201 [98 (0.490)], and the GA genotype frequency at K-ras gene locus rs1137188 [127 (0.635)] were all significantly higher than those in the control group (p=0.000, p=0.002, p=0.011). In the disease group, the frequency of recessive model GC+CC (p=0.003), heterozygous model GC (p=0.035), and homozygous model CC (p=0.037) at c-myc gene locus rs121918684 was significantly lower than that in the control group, and the frequency of recessive model CT+TT (p=0.046) at c-myc gene locus rs775522201 was also markedly lower than that in the control group. The haplotype frequency of c-myc CC (p=0.000), GC (p=0.000), and GT (p=0.018) in the disease group was different from that in the control group. Moreover, the CT genotype at c-myc gene locus rs775522201 was remarkably correlated with the c-myc gene expression, and the gene expression was markedly increased in the disease group. The TT genotype at K-ras gene locus rs12245 was correlated with the K-ras gene expression, and the gene expression was notably increased in the disease group. There was an association between GG genotype at c-myc gene locus rs121918684 and LDH level (p=0.000), between CT genotype at c-myc gene locus rs775522201 and PLT level (p=0.002), and between AA genotype at K-ras gene locus rs1137188 and Hb level (p=0.003). CONCLUSIONS: The c-myc and K-ras gene polymorphisms are associated with susceptibility to NHL, gene expression and levels of Hb, PLT, and LDH.

PTH promotes rabbit tibial fracture healing via the Notch signaling pathway.

OBJECTIVE: To explore the effect of parathyroid hormone (PTH) on the expression of Jagged1 in the rabbit tibial fracture healing, and its function and mechanism in this process via the Notch signaling pathway. MATERIALS AND METHODS: A total of 60 New Zealand white rabbits were randomly divided into control group (n=30) and experimental group (n=30). Then, a rabbit tibial fracture model was established. After surgery, the rabbits in experimental group were given 10 µg/kg PTH (1-34) once a day for 5 days a week, while those in control group were given an equal volume of normal saline. Six rabbits were randomly selected from each group at 1, 2, 3, 4, and 6 weeks after surgery to collect right tibia specimens. Next, X-ray examination, bone mineral density (BMD) test, histological detection, and serum biochemical test were performed. Additionally, the messenger ribonucleic acid (mRNA) expression levels of Notch1 and Jagged1 in the Notch signaling pathway were measured via polymerase chain reaction (PCR) assay. Their protein levels were detected through Western blotting analysis. RESULTS: The healing and BMD in experimental group were better than those in control group since cortical and medullary bridging was observed in the rabbits of experimental group at the 6th week after surgery. Plasma level of alkaline phosphatase (ALP), P content, and the product of Ca and P significantly increased (p<0.05) in experimental group. The pathological morphology of the calluses stained with hematoxylin-eosin (HE) in experimental group was overtly superior to that in control group. The PCR results revealed that both mRNA and protein levels of Notch1 and Jagged1 were lower in control group than those in experimental group (p<0.05). CONCLUSIONS: PTH (1-34) promotes the rabbit tibial fracture healing by regulating Jagged1 ligand molecules in the Notch signaling pathway.

Relationship between miR-375 regulating Ndrg2/IL-6/STAT3 signaling pathway and diabetic retinopathy in rats.

OBJECTIVE: To explore the relationship between micro ribonucleic acid (miR)-375 in regulating the N-Myc downstream-regulated gene 2 (Ndrg2)/interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) signaling pathway and diabetic retinopathy (DR) in rats. MATERIALS AND METHODS: Thirty Sprague- Dawley rats were randomly divided into Control group (n=10), Model group (n=10), and miR-375 inhibitor group [miR-375 small interfering RNA (siRNA) group, n=10]. The rats in Model group were injected with streptozotocin (STZ) via the tail vein to prepare into rat models of diabetes. The body weight, fasting blood glucose, and retinal barrier permeability of rats in each group were detected. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in rat serum were measured using kits. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay was performed to determine the apoptosis of optic ganglion cells in rat retinal tissues. Additionally, the messenger RNA (mRNA) and protein levels of Ndrg2, IL-6 and STAT3 in rat retinal tissues were detected via reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. RESULTS: Compared with Control group, Model group had reduced body weight of rats, increased blood glucose and retinal permeability of rats, raised serum MDA content, decreased SOD activity, up-regulated apoptotic rate of optic ganglia, and notably elevated mRNA and protein levels of Ndrg2, IL-6 and STAT3 in retinal tissues. Compared with those in Model group, the body weight of rats declined, the blood glucose of rats rose, the retinal permeability of rats was decreased significantly, the serum MDA content was reduced, the SOD activity was raised, the apoptotic rate of optic ganglia was decreased, and the mRNA and protein levels of Ndrg2, IL-6 and STAT3 in retinal tissues were also decreased significantly in miR-375 siRNA group. CONCLUSIONS: MiR-375 inhibitors are able to reduce blood glucose, retinal permeability, and optic ganglion apoptosis in rats with DR, and the mechanism of action may be related to the regulation on the Ndrg2/IL-6/STAT3 signaling pathway.

MiR-155-5p accelerates cerebral ischemia-reperfusion injury via targeting DUSP14 by regulating NF-κB and MAPKs signaling pathways.

OBJECTIVE: This study aimed to explore the role of miR-155-5p in middle cerebral artery occlusion/reperfusion (MCAO/R) model in rats and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced SH-SY5Y cells. In addition, this study also aimed to explore the underlying mechanisms to expect that miR-155-5p may be investigated as a new and effective diagnostic and therapeutic target for ischemic stroke. MATERIALS AND METHODS: The in vivo MCAO/R rat model and in vitro OGD/R cell model were established. The miR-155-5p mRNA expression was detected by quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Dual specificity ATPase (DUSP) 14 was predicted to be a potential target of miR-155-5p by TargetScan. The targeting relationship was confirmed by Luciferase assay. The cell viability was determined using the Cell Counting Kit-8 (CCK-8). The expression level of inflammatory cytokines, including tumor necrosis factor alpha (TNF-α), interleukin-1ß (IL-1ß), and interleukin-6 (IL-6) levels were detected by Enzyme-Linked Immunosorbent Assay (ELISA). Western blot was used to detect the protein expression of DUSP14, the apoptotic protein Cleaved cysteine-aspartic acid protease (caspase)-3, and Cleaved PARP, as well as nuclear factor kappa B (NF-κB) and MAPKs signaling pathways related proteins. RESULTS: MiR-155-5p was upregulated in both MCAO/R rats and OGD/R-induced SH-SY5Y cells. MiR-155-5p knockdown inhibited OGD/R-induced cell injury and inflammation, as well as MCAO/R-induced brain injury. MiR-155-5p regulated the NF-κB and MAPKs signaling pathways by targeting DUSP14. DUSP14 knockdown partially reversed the protective effect of miR-155-5p knockdown on OGD/R-induced SH-SY5Y cell injury and inflammation. CONCLUSIONS: MiR-155-5p accelerates cerebral I/R injury via targeting DUSP14 by regulating NF-κB and MAPKs signaling pathways. Inhibition of miR-155-5p significantly reduces apoptosis and brain injury. These results indicated that miR-155-5p plays a key role in cerebral I/R injury and has the potential to be explored as a new target for ischemic stroke.

Comb-locked cavity ring-down spectroscopy with variable temperature.

Temperature dependence of molecular absorption line shape is important information for spectroscopic studies and applications. In this work, we report a comb-locked cavity ring-down spectrometer employing a cryogenic cooler to perform absorption spectroscopy measurements at temperatures between 40 K and 300 K. As a demonstration, we recorded the spectrum of the R(0) line in the (2-0) band of HD at 46 K. The temperature was also confirmed by the Doppler width of the HD line. Spectra of CH4 near 1.394 µm were also recorded in a wide temperature range of 70-300 K. Lower-state energies of methane lines were analyzed by fitting these spectra, which can be directly compared with the HITRAN and TheoReTS databases. Considerable deviations were observed, indicating the need to investigate the assignments of the methane lines in this region.

Knockdown of thrombospondin 2 inhibits metastasis through modulation of PI3K signaling pathway in uveal melanoma cell line M23.

OBJECTIVE: Thrombospondin 2 (THBS2) expression and its prognostic value have been documented in several types of cancer. Nevertheless, the potential role and clinical significance of THBS2 in uveal melanoma (UM) have never been reported. Thus, in our study, we aimed to explore the clinical significance and prognostic impact of THBS2 in UM. MATERIALS AND METHODS: Survival and prognosis analyses were implemented using the Kaplan-Meier method and COX's proportional hazards model based on the clinical data retrieved from The Cancer Genome Atlas (TCGA) database. Colony formation, cell proliferation, invasion and migration assays in M23 cell line were performed to evaluate the effects of THBS2 on UM in vitro. To further reveal whether the dysregulated THBS2 expression regulates the UM metastasis, protein biomarkers including serine-threonine kinase (AKT), p-AKT, phosphoinositide 3-kinase (PI3K), p-PI3K, and p70S6K were measured using Western blotting analysis. RESULTS: THBS2 was up-regulated in metastatic UMs. Relationship of THBS2 expression level with the clinicopathological factors demonstrated that the expression level of THBS2 was significantly correlated to histological type, recurrence, and dead. Univariate as well as multivariate COX analyses demonstrated that THBS2 could serve as an independent prognostic factor for overall survival of UM. The knockdown of THBS2 significantly inhibited the proliferation rate of M23 cells, suppressed the colony numbers of M23 cells, lowered the invasive and migratory cell proportion. Importantly, Western blotting results implicated that THBS2 knockdown significantly decreased the expression level of p-AKT, p-PI3K, and p70S6K in M23 cell line. CONCLUSIONS: This is the first study to report that THBS2 may play important roles in UM progression and might be a novel prognosis biomarker for UM.

iTRAQ-based quantitative proteomic analysis of differentially expressed proteins in Litopenaeus vannamei in response to infection with WSSV strains varying in virulence.

The isobaric tags for relative and absolute quantitation (iTRAQ) were applied to identify differentially expressed proteins (DEPs) in Litopenaeus vannamei in response to different virulence white spot syndrome virus infection. A total of 2780 unique peptides corresponding to 754 proteins were identified. The number of significant differentially expressed proteins was 161, including 38 up-regulated ones and 123 down-regulated ones in low-virulence infection library compared with normal-virulence infection library. Gene Ontology function annotation indicated that the differentially expressed proteins mainly participated in the biological process. Subcellular location classification showed that the largest distribution of differentially expressed proteins was found in the cytoplasm in both down-regulated and up-regulated proteins. Kyoto encyclopaedia of genes and genomes analysis revealed that most of the differentially expressed proteins were involved in carbon metabolism. Moreover, three metabolic pathways, including carbon metabolism, inositol phosphate metabolism, and fructose and mannose metabolism, were significantly affected. Our findings offered a better understanding of the host response to different virulence white spot syndrome virus infection. SIGNIFICANCE AND IMPACT OF THE STUDY: White spot syndrome virus (WSSV) is one of the most harmful pathogens in shrimp farming. Previous studies have shown that genetic variations of white spot syndrome virus cause the strains of different virulence. The hosts also show different self-regulation when infected by different viruses. A better understanding of host response to white spot syndrome virus will help elucidate the virulence and pathogenesis of this unique pathogen.

Protective effects of fentanyl preconditioning on cardiomyocyte apoptosis induced by ischemia-reperfusion in rats.

We aimed to study the effect of fentanyl (Fen) preconditioning on cardiomyocyte apoptosis induced by ischemia-reperfusion (I/R) in rats. A total of 120 Sprague Dawley male rats (age: 3 months) were randomly divided into: sham operation group (S group), I/R group, normal saline I/R group (NS group), and fentanyl low, middle, and high dose groups (Fen1: 2 µg/kg; Fen2: 4 µg/kg; Fen3: 6 µg/kg). Heart rate (HR), mean arterial pressure (MAP), left ventricular developed pressure (LVDP), ±dp/dtmax, malondialdehyde (MDA), superoxide dismutase (SOD) activity, creatine phosphokinase-MB (CK-MB), and cardiac troponin-I (cTnI) were measured. Myocardial ischemic (MI) area, total apoptotic myocardial cells, and protein and mRNA expressions of B-cell lymphoma 2 (Bcl-2) and Bax were detected. HR and MAP were higher, while LVDP and ±dp/dtmax were close to the base value in the Fen groups compared to those in the I/R group. Decreased MDA concentration and CK-MB value and increased SOD activity were found in the Fen groups compared to the I/R group, while cTnI concentration was significantly lower in the Fen1 and Fen2 groups (all P<0.05). Myocardial damage was less in the Fen groups compared to the I/R group and the MI areas and apoptotic indexes were significantly lower in the Fen1 and Fen2 groups (all P<0.05). Furthermore, significantly increased protein and mRNA expressions of Bcl-2, and decreased protein and mRNA expressions of Bax were found in the Fen groups compared to the I/R group (all P<0.05). Fentanyl preconditioning may suppress cardiomyocyte apoptosis induced by I/R in rats by regulating Bcl-2 and Bax.

The relation of the serum aldosterone level and central serous chorioretinopathy - a pilot study.

OBJECTIVE: The aim of this study was to investigate the serum aldosterone level and abnormal levels of mineral corticoid in patients with the central serous chorioretinopathy (CSC). PATIENTS AND METHODS: All recruited patients with CSC received fundus fluorescein angiography (FFA), enhanced depth imaging spectral-domain optical coherence tomography (EDI-OCT) and serum aldosterone assay. The patients were classified into spontaneously resolved group and unresolved group according to a 3-months follow-up of Optical Coherence Tomography (OCT) examination. Patients from unresolved group were recruited to receive treatment with 40 mg spironolactone orally for 2 months. After the treatment, the EDI-OCT and best corrected visual acuity (BCVA) were performed again to assess the treatment efficacy. RESULTS: The study included 61 patients (72 eyes) with 34 patients in the unresolved group and 27 patients in the resolved group. The aldosterone level was significantly associated with the subfoveal choroidal thickness (SFCT) of revolved CSC eyes (r=0.342, p<0.05) as well as the SFCT of unresolved CSC eyes (r=0.348, p<0.05). And the aldosterone level in the unresolved CSC group was greater than that in the spontaneously resolved group (161.8 ± 50.1 ng/dl vs. 122.5 ± 50.5 ng/dl, p<0.05). The central macular thickness and SFCT were decreased significantly (p<0.05) after the treatment with 40 mg/d spironolactone for two months. CONCLUSIONS: The unresolved CSC patients were characterized by high level of aldosterone and thickened SFCT. Spironolactone treatment was associated with the improvement of chronic CSC. Besides, the side effect of spironolactone treatment was rare.

Protective effects of fentanyl preconditioning on cardiomyocyte apoptosis induced by ischemia-reperfusion in rats

We aimed to study the effect of fentanyl (Fen) preconditioning on cardiomyocyte apoptosis induced by ischemia-reperfusion (I/R) in rats. A total of 120 Sprague Dawley male rats (age: 3 months) were randomly divided into: sham operation group (S group), I/R group, normal saline I/R group (NS group), and fentanyl low, middle, and high dose groups (Fen1: 2 μg/kg; Fen2: 4 μg/kg; Fen3: 6 μg/kg). Heart rate (HR), mean arterial pressure (MAP), left ventricular developed pressure (LVDP), ±dp/dtmax, malondialdehyde (MDA), superoxide dismutase (SOD) activity, creatine phosphokinase-MB (CK-MB), and cardiac troponin-I (cTnI) were measured. Myocardial ischemic (MI) area, total apoptotic myocardial cells, and protein and mRNA expressions of B-cell lymphoma 2 (Bcl-2) and Bax were detected. HR and MAP were higher, while LVDP and ±dp/dtmax were close to the base value in the Fen groups compared to those in the I/R group. Decreased MDA concentration and CK-MB value and increased SOD activity were found in the Fen groups compared to the I/R group, while cTnI concentration was significantly lower in the Fen1 and Fen2 groups (all P<0.05). Myocardial damage was less in the Fen groups compared to the I/R group and the MI areas and apoptotic indexes were significantly lower in the Fen1 and Fen2 groups (all P<0.05). Furthermore, significantly increased protein and mRNA expressions of Bcl-2, and decreased protein and mRNA expressions of Bax were found in the Fen groups compared to the I/R group (all P<0.05). Fentanyl preconditioning may suppress cardiomyocyte apoptosis induced by I/R in rats by regulating Bcl-2 and Bax.

Kaempferol inhibited VEGF and PGF expression and in vitro angiogenesis of HRECs under diabetic-like environment

Diabetic retinopathy (DR) is one of the common and specific microvascular complications of diabetes. This study aimed to investigate the anti-angiogenic effect of kaempferol and explore its underlying molecular mechanisms. The mRNA expression level of vascular endothelial growth factor (VEGF) and placenta growth factor (PGF) and the concentrations of secreted VEGF and PGF were measured by qTR-PCR and ELISA assay, respectively. Human retinal endothelial cells (HRECs) proliferation, migration, and sprouting were measured by CCK-8 and transwell, scratching wound, and tube formation assays, respectively. Protein levels were determined by western blot. High glucose (25 mM) increased the mRNA expression levels of VEGF and PGF as well as the concentrations of secreted VEGF and PGF in HRECs, which can be antagonized by kaempferol (25 µM). Kaempferol (5-25 µM) significantly suppressed cell proliferation, migration, migration distance and sprouting of HRECs under high glucose condition. The anti-angiogenic effect of kaempferol was mediated via downregulating the expression of PI3K and inhibiting the activation of Erk1/2, Src, and Akt1. This study indicates that kaempferol suppressed angiogenesis of HRECs via targeting VEGF and PGF to inhibit the activation of Src-Akt1-Erk1/2 signaling pathway. The results suggest that kaempferol may be a potential drug for better management of DR.

Single nucleotide polymorphisms predisposing to asthma in children of Mauritian Indian and Chinese Han ethnicity

Our objective was to investigate the distributions of six single nucleotide polymorphisms (SNPs) MS4A2 E237G, MS4A2 C-109T, ADRB2 R16G, IL4RA I75V, IL4 C-590T, and IL13 C1923T in Mauritian Indian and Chinese Han children with asthma. This case-control association study enrolled 382 unrelated Mauritian Indian children, 193 with asthma and 189 healthy controls, and 384 unrelated Chinese Han children, 192 with asthma and 192 healthy controls. The SNP loci were genotyped using polymerase chain reaction (PCR)-restriction fragment length polymorphism for the Chinese Han samples and TaqMan real-time quantitative PCR for the Mauritian Indian samples. In the Mauritian Indian children, there was a significant difference in the distribution of IL13 C1923T between the asthma and control groups (P=0.033). The frequency of IL13 C1923T T/T in the Mauritian Indian asthma group was significantly higher than in the control group [odds ratio (OR)=2.119, 95% confidence interval=1.048-4.285]. The Chinese Han children with asthma had significantly higher frequencies of MS4A2 C-109T T/T (OR=1.961, P=0.001) and ADRB2 R16G A/A (OR=2.575, P=0.000) than the control group. The IL13 C1923T locus predisposed to asthma in Mauritian Indian children, which represents an ethnic difference from the Chinese Han population. The MS4A2 C-109T T/T and ADRB2 R16G A/A genotypes were associated with asthma in the Chinese Han children.

Prognostic value of clinical and immunological markers in acute phase of SFTS virus infection.

SFTS virus (SFTSV) is a novel bunyavirus that causes severe fever with thrombocytopenia syndrome (SFTS), an emerging infectious disease that occurred in China in recent years, with an average case fatality rate of 10-12%. Intervention in the early clinical stage is the most effective measure to reduce the mortality rate of disease. To elucidate the natural course of and immune mechanisms associated with the pathogenesis of SFTSV, 59 laboratory-confirmed SFTS patients in the acute phase, who were hospitalized between October 2010 and September 2011, were enrolled in this study, and the patients sera were dynamically collected and tested for SFTSV viral RNA load, 34 cytokines or chemokines and other related laboratory parameters. All clinical diagnostic factors in the acute phase of SFTS were evaluated and assessed. The study showed that the severity of the disease in 11 (18.6%) patients was associated with abdominal pain (p 0.007; OR = 21.95; 95% CI, 2.32-208.11) and gingival bleeding (p 0.001; OR=122.11; 95% CI, 6.41-2328). The IP-10, TNF-α, IL-6, IL-10, granzyme B and HSP70 levels were higher over the 7-8 days in severe cases, accompanied by altered AST, CK and LDH levels. HSP70 (p 0.012; OR=8.29; 95% CI, 1.58-43.40) was independently correlated with the severity of the early acute phase of SFTSV infection. The severity of SFTS can be predicted based on the presence of symptoms such as abdominal pain and gingival bleeding and on the level of HSP70 in the acute phase of the disease.

Osteogenesis induced in goat bone marrow progenitor cells by recombinant adenovirus coexpressing bone morphogenetic protein 2 and basic fibroblast growth factor

Bone morphogenetic protein 2 (BMP2) and basic fibroblast growth factor (bFGF) have been shown to exhibit a synergistic effect to promote bone repair and healing. In this study, we constructed a novel adenovirus with high coexpression of BMP2 and bFGF and evaluated its effect on osteogenic differentiation of goat bone marrow progenitor cells (BMPCs). Recombinant adenovirus Ad-BMP2-bFGF was constructed by using the T2A sequence. BMPCs were isolated from goats by density gradient centrifugation and adherent cell culture, and were then infected with Ad-BMP2-bFGF or Ad-BMP2. Expression of BMP2 and bFGF was detected by ELISA, and alkaline phosphatase (ALP) activity was detected by an ALP assay kit. In addition, von Kossa staining and immunocytochemical staining of collagen II were performed on BMPCs 21 days after infection. There was a high coexpression of BMP2 and bFGF in BMPCs infected with Ad-BMP2-bFGF. Twenty-one days after infection, ALP activity was significantly higher in BMPCs infected with Ad-BMP2-bFGF than in those infected with Ad-BMP2. Larger and more mineralized calcium nodules, as well as stronger collagen II staining, were observed in BMPCs infected with Ad-BMP2-bFGF than in those infected with Ad-BMP2. In summary, we developed a novel adenovirus vector Ad-BMP2-bFGF for simultaneous high coexpression of BMP2 and bFGF, which could induce BMPCs to differentiate efficiently into osteoblasts.

Randomized prospective study of olecranon fracture fixation: cable pin system versus tension band wiring.

OBJECTIVE: This prospective, randomized study compared the effectiveness of the cable pin system (CPS) versus tension band wiring (TBW) for olecranon fracture fixation. METHODS: Patients with acute transverse or slight oblique olecranon fractures were randomly divided into two groups: one fixed by CPS and the other by TBW. Clinical outcome data were collected and analysed following a mean duration of 21 months. RESULTS: The mean ± SD fracture healing time was significantly shorter in the CPS group (n = 30; 9.73 ± 2.02 weeks) compared with the TBW group (n = 32; 11.13 ± 2.21 weeks). One patient in the CPS group and seven patients in the TBW group experienced postoperative complications; this difference was statistically significant. The mean ± SD Mayo Elbow Performance Score in the CPS group was significantly higher (88.67 ± 6.42) than that in the TBW group (80.78 ± 11.99). Logistic regression analysis showed an association between fixation method and fracture healing time, complications and elbow function. CONCLUSIONS: Internal fixation by CPS is an effective method for olecranon fracture and is associated with a shorter healing time, fewer complications and better function than TBW.

Value of a skin island flap as a postoperative predictor of vascularized fibula graft viability in extensive diaphyseal bone defect reconstruction.

PURPOSE: To evaluate the feasibility and reliability of free vascularized fibular graft with skin island flap for reconstruction of large diaphyseal bone defect. METHOD: The clinical results of vascularized fibular graft and experiences related to the importance and reliability of a monitoring island flap for the reconstruction of various long-bone defects were reviewed in 87 patients. RESULTS: Bony reconstruction was achieved in 82 of the 87 patients. Arterial thrombosis of anastomosed vessel in two patients and venous congestion of monitoring flap in nine patients occurred in the early postoperative periods. All of them were managed by immediate thrombectomy and reanastomosis, alternatively the thrombotic veins were replaced by new veins to anastomose with the superficial veins in five patients. Partial flap necrosis was noted in six patients, but additional surgical intervention was not required. The vascularized fibula survived and bony fusion was achieved in all patients. Postoperative stress fractures of the fibula graft occurred in 19 (21.8%) patients (once in seven patients, twice in five patients, three or more times in seven) as the mechanical stress to the graft increased. Included fracture on the tibia in 12 patients, humerus in one and femur in six. Treatments included casting in 11 patients, percutaneous pinning in one case, and adjustment of external fixator in seven patients. Bony union was finally achieved an average of 9.6 months after fracture. CONCLUSIONS: Correct alignment between the recipient bone and the external fixator is a prerequisite to preventing graft fracture. Vascularized fibula transfer is a valuable procedure for long-bone defects, and a skin island-monitoring flap is a simple, extremely useful, and reliable method for assessing the vascular status of vascularized fibula. LEVEL OF EVIDENCE: Level IV. Retrospective study.

Enhanced cellular immune response elicited by a DNA vaccine fused with Ub against Mycobacterium tuberculosis.

This study evaluated the immune response elicited by a Ub-fused Ag85A DNA vaccine against Mycobacterium tuberculosis. BALB/c mice were vaccinated with plasmid DNA encoding Ag85A protein, Ub-fused Ag85A DNA vaccine (UbGR-Ag85A) and negative DNA vaccines, respectively. Ag85A DNA vaccine immunization induced a Th(l)-polarized immune response. The production of Th(l)-type cytokine (IFN-γ) and proliferative T cell responses was enhanced significantly in mice immunized with UbGR-Ag85A fusion DNA vaccine, compared with non-fusion DNA vaccine. Moreover, this fusion DNA vaccine also resulted in an increased relative ratio of IgG(2a) to IgG(l) and the cytotoxicity of T cells. IFN-γ intracellular staining of splenocytes indicated that UbGR-Ag85A fusion DNA vaccine activated CD4(+) and CD8(+) T cells, particularly CD8(+) T cells. Thus, this study demonstrated that the UbGR-Ag85A fusion DNA vaccine inoculation could improve antigen-specific cellular immune responses, which is helpful for protection against TB infection.

VP37 of white spot syndrome virus interact with shrimp cells.

AIMS: To investigate VP37 [WSV 254 of White spot syndrome virus (WSSV) genome] interacting with shrimp cells and protecting shrimp against WSSV infection. METHODS AND RESULTS: VP37 was expressed in Escherichia coli and was confirmed by Western blotting. Virus overlay protein binding assay (VOPBA) technique was used to analyse the rVP37 interaction with shrimp and the results showed that rVP37 interacted with shrimp cell membrane. Binding assay of recombinant VP37 with shrimp cell membrane by ELISA confirmed that purified rVP37 had a high-binding activity with shrimp cell membrane. Binding of rVP37 to shrimp cell membrane was a dose-dependent. Competition ELISA result showed that the envelope protein VP37 could compete with WSSV to bind to shrimp cells. In vivo inhibition experiment showed that rVP37 provided 40% protection. Inhibition of virus infection by rVP37 in primary cell culture revealed that rVP37 counterparted virus infection within the experiment period. CONCLUSIONS: VP37 has been successfully expressed in E. coli. VP37 interacted with shrimp cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that rVP37 has a potential application in prevention of virus infection.

A single-step multiplex PCR for simultaneous detection of white spot syndrome virus and infectious hypodermal and haematopoietic necrosis virus in penaeid shrimp.

White spot syndrome virus (WSSV) and infectious hypodermal and haematopoietic necrosis virus (IHHNV) are the major viral pathogens of penaeid shrimp worldwide (Lightner & Redman 1998). Litopenaeus vannamei was introduced into China from the Americas, and quickly became widely cultured. Following its introduction, both IHHNV and WSSV have become important pathogens of cultured penaeid shrimp and have had a huge impact on the culture industry in China in recent years.