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Viral-encoded microRNAs (miRNAs) have vital roles in the regulation of virus replications and host immune responses. The results of previous studies have indicated that miRNA clusters are involved in the replication and virulence of the pseudorabies virus (PRV), which may potentially lead to immune escape or facilitation of PRV replication. This study's previous research revealed that prv-miR-LLT11a was differentially expressed during PRV infection. The present study's results have demonstrated that prv-miR-LLT11a could significantly inhibit PRV replication. It was further determined that SLA-1 was the target gene of prv-miR-LLT11a, and simultaneously, that overexpression of prv-miR-LLT11a could downregulate the mRNA and protein levels of SLA-1 in a dose-independent manner. Furthermore, the present study also observed that prv-miR-LLT11a can downregulate TAP1 expression. Our findings provide a better understanding of the molecular mechanism involved in the effects of prv-miR-LLT11a on SLA-1 and TAP1 as well as its involvement in immune system evasion of PRV.
Subject(s)
Herpesvirus 1, Suid , Immune System , MicroRNAs , Pseudorabies , RNA, Messenger , United Nations , Virulence , Virus ReplicationABSTRACT
Objective: To compare the short-term effectiveness between arthroscopic cystectomy and internal drainage combined with cystectomy in popliteal cyst. Methods: Between March 2014 and March 2017, 56 patients with symptomatic popliteal cyst were enrolled in the study, randomized block design was used to divided the patients into trial group (arthroscopic cystectomy combined with internal drainage group, n=28) and control group (arthroscopic internal drainage group, n=28). Excluding those who had incomplete follow-up and received surgery for other diseases postoperatively, 26 patients in the experimental group and 27 patients in the control group were finally enrolled in the study. There was no significant difference in gender, age, side, course of disease, maximum diameter and grade of popliteal cyst, and associated diseases between two groups ( P>0.05). The operation time, duration of popliteal ecchymosis and the middle back of calf tenderness were observed postoperatively. The circumference of calf at 1 day, 1 week, and 2 weeks after operation were measured and the differences were calculated with the measurement before operation. Lower extremity venous thrombosis was observed by color doppler ultrasonography at 1 week after operation. The effectiveness was evaluated by Rauschning and Lindgren grading criteria. And MRI was used to observe whether the popliteal cyst disappeared or decreased and measured its maximum diameter at 1 year after operation. Results: Patients in both groups were followed up 12-14 months, with an average of 12.5 months. The operation time, duration of popliteal ecchymosis, and the middle back of calf tenderness of the trial group were all longer than those in the control group ( P<0.05), the differences of circumference of calf at 1 day, 1 week, and 2 weeks after operation of the trial group were greater than those in the control group ( P<0.05). Color doppler ultrasonography of the lower extremity at 1 week after operation found that the intermuscular venous thrombosis occurred in 2 cases of the trial group, while no lower extremity thrombosis was found in the control group; and the difference between two groups was not significant ( P=0.236). According to the Rauschning and Lindgren grading criteria, there were 16 cases of grade 0, 6 cases of grade 1, and 4 cases of grade 2 in the trial group, and 17 cases of grade 0, 4 cases of grade 1, and 6 cases of grade 2 in the control group at 1 year after operation. There was no significant difference between 2 groups ( Z=-1.872, P=0.078). Nine cases (34.62%) of the trial group and 13 cases (48.15%) of the control group still have residual cysts by MRI, the maximum diameter of which was less than 2 cm. The cysts disappeared in the remaining patients in both groups, and there was no recurrence during the follow-up. There was no significant difference in cyst residual rate between 2 groups ( χ2=2.293, P=0.852). Conclusion: Compared with arthroscopic internal drainage, the short-term effectiveness of the arthroscopic internal drainage combined with cystectomy had no significant improvement, and the operation time was prolonged, the postoperative complications were obviously increased.
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Objective To identify and assess whether circumferential electrocautery is useful to improve outcomes after primary total knee replacement.Methods We searched MEDLINE,EMBASE,PubMed,SpringerLink,Web of Knowledge,OVID CINAHL,OVID EBM and Google Scholar,and included articles published through January 2014.This study used the methods of meta-analysis to assess whether circumferential electrocautery could improve anterior knee pain and knee function after total knee replacement,and identified the function of circumferential electrocautery in total knee replacement.Results A total of eight articles met the inclusion criteria.Among the 1 048 cases included in the analysis,526 cases had patellar denervation,and 522 cases were in the control group.The meta-analysis revealed no significant difference in the incidence of anterior knee pain (P =0.18) and visual analogue scale (VAS) (P =0.23) between two groups.However,patellar score (P =0.01),Western Ontario and McMaster Universities osteoarthritis index (WOMAC) score (P ＜ 0.01),and range of motion (ROM) (P ＜ 0.01) indicated that circumpatellar electrocautery improved clinical outcomes compared to non-electrocautery groups.Conclusions The results indicated that circumferential electrocautery of the patellar did not significantly improve anterior knee pain relative to non-electrocautery techniques.However,circumferential electrocautery significantly improved patients' knee function after surgery.Therefore,circumferential electrocautery might be beneficial to the outcome of primary total knee replacement surgery without patella replacement.
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Objective To investigate the repair effects of co-expression of the VEGF and BMP genes via an adeno-as-sociated viral vector on early steroid-induced avascular necrosis of the femoral head in rabbits. Methords To construct ani-mal model of early SANFH and screen by MRI. The SANFH animal were divided into rAAV-IRES-hrGFP(AAV-GFP), rAAV-hVEGF165-IRES-hrGFP(AAV-VEGF), rAAV-hBMP-7-IRES-hrGFP(AAV-BMP)and rAAV-hVEGF165-IRES-hBMP-7(AAV-VEGF/BMP)groups. The four group virus vectors were injected into core decompression region at the dose of 25μl/site after core decompression operation directly. Repair effects of rAAV vector on early SANFH in rabbits were evaluated by Western blot assay, HE staining, immunohistochemical staining, MRI, radionuclide bone scan, blood vessel counting detected by ink perfusion and fro-zen section, Micro-CT and biomechanical strength detection on the 12th week post-injection. Results Model success ratio was 73.33%. rAAV-hVEGF165-IRES-hBMP-7 virus vector efficiently expressed hVEGF165 and hBMP-7 genes on the 12th week after rAAV injection. hVEGF165 protein secreted in vivo promoted metabolism in core decompression region by increasing the quantity of new vessels and improving the blood supply;hBMP-7 protein secreted in vivo promoted new bone formation in core decompres-sion region by increasing bone mineral density and improving bone biomechanical strength. The AAV-VEGF/BMP group can pro-mote repair effects more effectively than AAV-VEGF group or AAV-BMP group. Conclusion The adeno-associated viral vectors co-expressing hVEGF165 and hBMP-7 can promote repair effects on early SANFH in rabbits by increasing the blood supply and strengthening the bone quality of femoral head.
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Objective To investigate the serum levels of β-amyloid precursor protein (β-APP) and hypersensitive-cross-reactive protein(hs-CRP) after acute spinal cord injury(SCI) in rats at different time points and study their roles in secondary spinal cord injury. Methods Ninety healthy adult female SD rats were randomly assigned into 3 even groups.Improved Allen method was used to create models of complete SCI (group A) and models of incomplete SCI (group B).In group C only laminectomy was performed without SCI.The serum was collected at 12 hours,24 hours,3 days,7 days,and 14 days after operation through femoral vein to detect levels of β-APP and hs-CRP by ELISA. ResultsThe serum level of β-APP reached the peak value 24 hours after acute SCI,and the serum level of hs-CRP reached the peak 3 days after acute SCI.There were significant differences between groups A and C in β-APP and hs-CRP levels at all time points( P ＜ 0.05).There were also significant differences between groups B and C in β-APP and hs-CRP levels at all time points (P ＜0.05) except at 14 days (P＞ 0.05). Conclusions The serum levels of β-APP and hs-CRP are positively associated with the severity of traumatic SCI.β-APP may be a more sensitive indicator than hs-CRP in a certain time.The dynamic changes of serum β-APP and hs-CRP suggest involvement of the 2 proteins in the secondary spinal cord injury.
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Objective To investigate the effects of Panax notoginseng saponins (PNS) on hydrogen peroxide (H2O2)-induced apoptosis in cultured rabbit bone marrow stromal cells (BMSCs). Methods BMSCs from 3-month-old New Zealand rabbits were isolated and cultured by the density gradient centrifugation combined with adherent method. The cultured BMSCs were divided into three groups: normal control, H2O2 treatment (100μmol/L), and PNS pretreatment (0.1g/L). Intracellular reactive oxygen species (ROS) levels as the index of oxidative stress were measured by using 2'7'-dichlorodihydrofluorescein diacetate. Flow cytometry was used to observe the apoptosis of BMSCs by staining with annexinV-FITC/PI. The protein expression of Bax in BMSCs was analyzed by Western blotting. Activity of caspase-3 enzyme was measured by spectrofluorometry. Results Pretreatment with PNS significantly decreased intracellular ROS level induced by H2O2 (P<0.01). PNS markedly attenuated H2O2-induced apoptosis rate from 38.68% to 19.24%(P<0.01). PNS reversed H2O2-induced augmentation of Bax expression. Furthermore, PNS markedly reduced the altered in activity of caspase-3 enzyme induced by H2O2(P<0.01). Conclusion PNS has a protective effect on hydrogen peroxide-induced apoptosis in cultured rabbit BMSCs by scavenging ROS and decreasing Bax expression and caspase-3 activity.
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Objective To observe the effect of the in vitro isolation, culture and identification of rabbit bone marrow-derived mesenchymal stem cells (BMSCs) and explore the differentiation potentials of BMSCs. Methods BMSCs were isolated and purified by density gradient centrifugation combined with attachment culture method. The biological characteristics of BMSCs were observed under an inverted microscope. The BMSCs were identified with HE staining and CD34/CD44 immunohistochemical staining. The biological activity of BMSCs was detected by osteogenic induction and steatogenic induction. Results BMSCs were isolated successfully in vitro and had good homogenicity. Immunohistochemical staining of CD34/CD44 showed strong positive expression of CD44 and negative expression of CD34. Fortis osteogenic ability and positive staining of ALP could be detected by osteogenic induction, and fortis steatogenic ability and positive staining of axungia could be detected by steatogenic induction. Conclusion BMSCs have the capability of multi-directional differentiation in vitro.
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ts the shear stress, thus, facilitates the union of fracture and the restoration of function.
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Objective To evaluate the effect of alendronate on osteoprotegerin (OPG) and receptor of activator of nuclear factor κB-ligand (RANKL) expression in human marrow stroma cells (hMSCs) in vitro. Methods hMSCs were isolated from haman marrow, cultured in vitro, and randomly divided into two groups: alendronate group, hMSCs culture fluid containing 1×10-7 mol/ L alendronate; control group, no special treatment but culturing hMSCs in DMEM. Two weeks after treatment, the expressions of OPG and RANKL were evaluated by RT-PCR and Western blot.Results hMSCs became uniform spindle-shaped fibroblasts. As cells proliferated, they formed colonies and showed whirlpool arrangement. After one week's treatment, hMSCs in alendronate group had reduced processes and gradually showed disc shape, which did not happen in control group but kept fibroblast shape and just increased in density. In RT-PCR, the ratio of OPG/RANKL in alendronate group and control group was 8.77±1.16 and 4.58±1.27, respec-tively. In Western blot, the ratio of OPG/RANKL in alendronate group and control group was 2.58±0.47 and 1.52±0.32, respectively. The ratio of OPG/RANKL was higher in alendronate group than in control group (P<0.01).Conclusion Alendronatc enhances OPG expression and inhibits RANKL expression of hMSCs in vitro.
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To explore the effects of different doses of puerarin on proliferation of cultured vascular endothelial cells in vitro.
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Objective To compare the clinical effects of treating cervical prolapse of intervertebral disc by using Solis cage, titanium cage and autogenous iliac crest graft (AICG) combined with titanium plate. Methods 64 cases of cervical prolapse of intervertebral disc were analyzed retrospectively. All the patients were followed up for about one year. Group A is composed of 20 patients (30 intervertebral spaces) treated with microdiscectomy and Solis cage fusion;group B is composed of 21 patients(22 intervertebral spaces) treated with titanium cage and group C is involved of 23patients(28 intervertebral spaces) which were dealt with AICG combined with titanium plate. The differences in the to-tal X ray exposure time, time working on iliac bone, incidence of complications, fusion rate, incidence of JOA score recovery rate more than 50 % and rehabilitation time of the three groups are analyzed. Results All indexes from group A and B were more satisfactory than those from group C (P＜0. 05). The statistic analysis results of the incidence of complications of cervical part, rehabilitation time in group A are 5.0± 1.8, 5.1 ± 1.2; and that is 14.3±2.6, 7.5 ± 1.6and 26.1±6.2, 8.6±2.3 in group B and group C respectively. There is significant difference between group A and group B on the incidence of complications of cervical part and rehabilitation time (P＜0.05). Results of the incidence of JOA score recovery rate ＞ 50 % in group A(95.8 ± 3.2) was more satisfactory than that in group B(93.6 ± 2.9),while there was no statistical difference between group A and group B (P＞0.05). Conclusion The Solis cage is better in treating the cervical prolapse of intervertebral disc than other ways. Also, it makes the operation easier, rehabilitation time shorter, and it causes fewer complications.
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OBJECTIVE: To observe the promoting effect of arnebia root oils on expression of basic fibroblast growth factor (bFGF) in skin wound of rabbits and the histomorphological changes in the wound surface, and to discuss its mechanism. METHODS: Bilateral round skin wounds were made on the back of 15 rabbits. The three wounds on one side of the back of each rabbit were treated with arnebia root oils, while the three wounds on the other side were treated with vaseline in order to promote the wound healing. The histomorphology and ultrastructure under electron microscopy of the wounds, and the rate of wound healing were examined at different time. Western blotting assay was used to detect the expression of bFGF in the wound surface. RESULTS: The healing rate of the arnebia root oils-treated wounds was evidently higher than that of the vaseline-treated wounds (P<0.05). The quantities of fibroblast, collagen and capillary in the arnebia root oils-treated wounds were much more than those in the vaseline-treated wounds, and the expression of endogenous bFGF in the arnebia root oils-treated wounds was enhanced obviously as compared with that in the vaseline-treated wounds in different period of wound healing. There existed a parallel correlation between the expression level of bFGF and the rate of wound healing. CONCLUSION: The promoting effect of arnebia root oils on wound healing may be related to increasing the expression level of basic fibroblast growth factor in the skin wound.
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<p><b>OBJECTIVE</b>To explore the molecular biological mechanism of Arnebia Root oil (AO) in promoting the recovery of surface of wound by observing basic fibroblast growth factor (bFGF) mRNA expression in the wound tissue and healing rate of the wound.</p><p><b>METHODS</b>Patients in the observed group with their wound treated by AO and those in the control group treated by petrolatum gauze. The wound surface healing rate was estimated and bFGF mRNA expression was observed by RT-PCR.</p><p><b>RESULTS</b>Endogenous bFGF mRNA expression existed in the wound surface of both groups, but its level in the observed group at any time point was obviously higher than that in the control group respectively, with significant difference in comparison of the gray density between the two groups (P < 0.05). The wound surface healing rate kept abreast with bFGF mRNA expression in wound tissues, so it was significantly higher in the observed group than that in the control group (P < 0.05). GAPDH gene, which was taken as a parameter for internal reference, expressed with a certain amount unchanged in different periods of healing (P > 0.05 ).</p><p><b>CONCLUSION</b>AO shows obviously promotive action on bFGF, an important regulatory factor on wound healing, it might complete the recovery process by stimulating the increase of bFGF.</p>