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BACKGROUND: Nano-hydroxyapatite/polyamide 66 (nHA/PA66) is a composite used widely in the repair of bone defects. However, this material is insufficient bioactivity. In contrast, D-RADA16-RGD self-assembling peptide (D-RADA16-RGD sequence containing all D-amino acids is Ac-RADARADARADARADARGDS-CONH2) shows admirable bioactivity for both cell culture and bone regeneration. Here, we describe the fabrication of a favorable biomaterial material (nHA/PA66/D-RADA16-RGD). METHODS: Proteinase K and circular dichroism spectroscopy were employed to test the stability and secondary structural properties of peptide D-RADA16-RGD respectively. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to characterize the surface of these materials. Confocal laser scanning (CLS), cell counting kit-8 tests (CCK-8), alizarin red S staining, cell immunofluorescence analysis and Western blotting were involved in vitro. Also biosafety and bioactivity of them have been evaluated in vivo. RESULTS: Proteinase K and circular dichroism spectroscopy demonstrated that D-RADA16-RGD in nHA/PA66 was able to form stable-sheet secondary structure. SEM and TEM showed that the D-RADA16-RGD material was 7–33 nm in width and 130–600 nm in length, and the interwoven pore size ranged from 40 to 200 nm. CLS suggests that cells in nHA/PA66/D-RADA16-RGD group were linked to adjacent cells with more actin filaments. CCK-8 analysis showed that nHA/PA66/D-RADA16-RGD revealed good biocompatibility. The results of Alizarin-red S staining and Western blotting as well as vivo osteogenesis suggest nHA/PA66/D-RADA16-RGD exhibits better bioactivity. CONCLUSION: This study demonstrates that our nHA/PA66/D-RADA16-RGD composite exhibits reasonable mechanical properties, biocompatibility and bioactivity with promotion of bone formation.
Subject(s)
Actin Cytoskeleton , Blotting, Western , Bone Regeneration , Cell Count , Cell Culture Techniques , Circular Dichroism , Endopeptidase K , Fluorescent Antibody Technique , In Vitro Techniques , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Osteogenesis , Sincalide , Spectrum AnalysisABSTRACT
Currently,there is no study on the unified coding of the spinal nerve and its main branches.The positions of spinal nerve were encoded according to the basic anatomical principles from top to bottom,from inside to outside and from front to back,with reference to AO bone classification and somatic artery coding and injury classification system.The segmental coding of spinal nerves was decided by its branches and running characteristics.The spinal nerve injuries were encoded by a combination of numbers and letters.The first number represented the region,the second represented the injured nerve,and the third represented the specific segment of injured nerve.The injuries of spinal nerve were divided into 5 categories according to severity.The first letter indicated the category of injury,and the second indicated the orientation of the injured nerve.The functional score scale of the innervation area was prepared based on the evaluation of motor function,sensory function and the results of neurophysiological examination.This scale was used to classify the neurological injuries,guide the clinical treatment and evaluate the prognostic outcomes.This coding and classification system can clearly and comprehensively describe the location and type of spinal nerve injuries,and it is convenient for the diagnosis of nervous system damage.It also has important reference value for treatment and prognosis evaluation.
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Objective To screen stable express A30P a-synuclein PC12 cell as the research object,and to preliminarily investigate the activity of different glycogen systhesis kinase 3β (GSK3β) for the regulation of autophagy pathway causes a-synuclein autophagy level change and its effects on cell growth.Methods The vector of A30P a-synuclein was used to transfect to PC12 cells.G418 was used to screen stable expressed cells.Liposome method was used to transfect GSK3β-expressed plasmid,GSK3β silence plasmid,and blank plasmid into these cell lines,and stable expressed activity of GSK3β cell lines were screened.Western blot was used to detect the expression of GSK3 β after transfection and verify the transfection efficiency.Western blot was used to test groups of cells in LC-3 Ⅱ,Beclin1,and a-synuclein expressions.Methyl thiazolyl tetrazolium (MTF) method was used to detect the change of cell proliferation.Terminal dexynucleotidyl transferase (TdT) mediated dUTP nick end labeling (TUNEL) method was used to detect apoptosis.Results (1) When GSK3β expressed group was compared to GSK3β silence group,blank plasmidgroup,and blank control group,the expressions of LC-3 Ⅱ and Beclin1 were significantly increased (P ＜ 0.05),the expression of a-synuclein was significantly reduced (P ＜ 0.05),the results of MTT and TUNEL showed that the cells were the decrease of apoptosis and increase of cell proliferation,with a statistically significant difference (P ＜ 0.05).(2) When GSK3β silence group was compared to GSK3β express group,blank plasmidgroup,and blank control group,the expressions of LC-3 Ⅱ and Beclin1 were significantly reduced (P ＜ 0.05),the expression of a-synuclein was significantly increased (P ＜ 0.05),the results of MTT and TUNEL showed that the cells were the increase of apoptosis and decrease of cell proliferation,with a statistically significant difference (P ＜0.05).Conclusions (1) Activation of GSK3β activity can improve the level of cell autophagy,enhance the ability of alpha-synuclein degradation,and promote cell survival.(2) Inhibition of GSK3β activity can reduce the level of cell autophagy,weaken the ability of alpha-synucleindegradation,and reduce cell survival.(3) Autophagy is closely related to the pathogenesis of Parkinson's disease (PD).The improvement of the level of autophagy and enhancing of the degradation of asynuclein is a new way of the future treatment of PD.
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Objective To screen stable express A30P a-synuclein PC12 cell as the research object,and to preliminarily investigate the activity of different glycogen systhesis kinase 3β (GSK3β) for the regulation of autophagy pathway causes a-synuclein autophagy level change and its effects on cell growth.Methods The vector of A30P a-synuclein was used to transfect to PC12 cells.G418 was used to screen stable expressed cells.Liposome method was used to transfect GSK3β-expressed plasmid,GSK3β silence plasmid,and blank plasmid into these cell lines,and stable expressed activity of GSK3β cell lines were screened.Western blot was used to detect the expression of GSK3 β after transfection and verify the transfection efficiency.Western blot was used to test groups of cells in LC-3 Ⅱ,Beclin1,and a-synuclein expressions.Methyl thiazolyl tetrazolium (MTF) method was used to detect the change of cell proliferation.Terminal dexynucleotidyl transferase (TdT) mediated dUTP nick end labeling (TUNEL) method was used to detect apoptosis.Results (1) When GSK3β expressed group was compared to GSK3β silence group,blank plasmidgroup,and blank control group,the expressions of LC-3 Ⅱ and Beclin1 were significantly increased (P ＜ 0.05),the expression of a-synuclein was significantly reduced (P ＜ 0.05),the results of MTT and TUNEL showed that the cells were the decrease of apoptosis and increase of cell proliferation,with a statistically significant difference (P ＜ 0.05).(2) When GSK3β silence group was compared to GSK3β express group,blank plasmidgroup,and blank control group,the expressions of LC-3 Ⅱ and Beclin1 were significantly reduced (P ＜ 0.05),the expression of a-synuclein was significantly increased (P ＜ 0.05),the results of MTT and TUNEL showed that the cells were the increase of apoptosis and decrease of cell proliferation,with a statistically significant difference (P ＜0.05).Conclusions (1) Activation of GSK3β activity can improve the level of cell autophagy,enhance the ability of alpha-synuclein degradation,and promote cell survival.(2) Inhibition of GSK3β activity can reduce the level of cell autophagy,weaken the ability of alpha-synucleindegradation,and reduce cell survival.(3) Autophagy is closely related to the pathogenesis of Parkinson's disease (PD).The improvement of the level of autophagy and enhancing of the degradation of asynuclein is a new way of the future treatment of PD.
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Ischemic cerebrovascular disease is a common and frequently occurring disease in the elderly.It has the characteristics of high disability rate,high mortality rate,and high recurrence rate,which seriously threatens the life and health of the elderly.At present,clinical diagnosis and evaluation are mainly based on medical history,neurological symptoms and signs and imaging examination.The incidence of ischemic cerebrovascular disease is rapid,the existing diagnostic methods have different degrees of hysteresis and limitations in the diagnosis time.Related clinical study found that in the process of ischemic cerebrovascular disease accompanied by serum S-100 beta,matrix metalloproteinase-9 (MMP-9)-related indicators of change.This article reviews the recent progress of S-100 beta and MMP-9 in ischemic cerebrovascular disease.
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Objective:To optimize the ultrasonic extraction process of Danggui Baogan capsules. Methods:The ultrasonic extrac-tion process of Danggui Baogan capsules was optimized by L9 (34 ) orthogonal test with the contents of ferulic acid and angelica sinensis polysaccharide as the indices and the amount of 30% ethanol, ultrasonic duration and extraction times as the influencing factors. Re-sults:The best ultrasonic extraction conditions were as follows: adding 12-fold amount of 30% ethanol and extracted four times with 1. 5 hours for each time. Conclusion:The ultrasonic extraction process is simple and reproducible, and suitable for the industrial pro-duction.
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Objective:To observe the clinical efficacy of interaction acupuncture combining with routine acupuncture for intractable facial palsy. Methods:A total of 60 eligible cases were randomly allocated into a treatment group (n=30) and a control group (n=30). Cases in the treatment group received interaction and routine acupuncture, whereas cases in the control group received routine acupuncture alone. The treatment was done once a day and 10 times made up a course of treatment. The patients were treated for a total of 3 courses and there were no intervals between two courses. Results:The total effective rate was 93.3% in the treatment group (including 17 recovery cases, 11 improvement cases and 2 failure cases), versus 76.7% in the control group (including 8 recovery cases, 15 improvement cases and 7 failure cases), showing a statistical difference (P Conclusion:Combining interaction and routine acupuncture can obtain better effect than routine acupuncture alone for intractable facial palsy.
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OBJECTIVE@#To explore the efficacy of the cognitive behavior therapy (CBT) for the treatment of chronic subjective tinnitus.@*METHOD@#One hundred and fifty-seven patients were randomly divided into two groups. Sixty-eight patients of the control group were treated by masking therapy; and the other 89 patients of the experimental group were treated by CBT therapy. The score of tinnitus handicap inventory (THI) was utilized to analyze the treatment efficacy in the two groups respectively.@*RESULT@#The effective rate assessed by of THI score in the experimental group was not significantly higher than the control group 2 months after treatment (P > 0.05), but was significantly higher than the control group 6 months and 12 months after treatment (P < 0.05 respectively).@*CONCLUSION@#The CBT therapy contributed to achieve rapid adaptation of tinnitus feeling, which shows great value of further clinical application.
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Humans , Cognitive Behavioral Therapy , Tinnitus , Psychology , Therapeutics , Treatment OutcomeABSTRACT
ObjectiveTo determine the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphism in Han ethnic patients with type 2 diabetes in Chenzhou area of Hunan province,and to assess the assoeiation between this polymorphism and diabetes.MethodsThis study was cross-sectional designed.Samples were taken by random sampling method from diabetes, prediabetes and euglycemia subjeers.ACE gene polymorphism I/D were examined by PCR.Genotype was classified as I1, ID, or DD based on positive or negative insertion/deletion allele.ResultsThis study showed significant differences of ages (X2 =32.09, P ＜0.01), body mass index (BMI) (X2 =16.10, P ＜0.01), family history (X2 =20.42, P ＜0.01) and serum triglyceride (F =21.289, P ＜0.01) among three groups, and diabetes group tended to have higher value.Frequency of genotype II, ID, DD were 72(56.3％) ,52(40.6％) and 4(3.1％) in diabetes, 64(64.0％) ,30(30.0％) and 6(6.0％) in pre-diabetes subjects, 81 (72.3％),29(25.9％)和 2(1.8％) in euglycemia subjects, consecutively.Chi-square analysis showed statistically significant association between ID + DD vs.Ⅱ genotype and diabetes (x2 = 6.68, P ＜ 0.05).Logistic regression analysis showed four variables that significantly influence to diabetes, namely ACE gene polymorphism, family history of diabetes, BMI, and serum triglyceride level.ConclusionsACE ID + DD genotype had significant relationship with diabetes in Chenzhou Han ethnic population, Hunan province, China.
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<p><b>OBJECTIVE</b>To investigate the relationship between G1025C (Try316Ser) polymorphism in exon 8 of apolipoprotein H (apoH) gene and stroke and to evaluate the effect of G1025C(Try316Ser) polymorphism on plasma lipid levels in Changsha Hans.</p><p><b>METHODS</b>G1025C (Try316Ser) polymorphism in apoH gene was determined by PCR-single strand conformation polymorphism analysis and DNA sequencing in 100 healthy controls, 260 patients with stroke, and 20 stroke pedigrees. Serum antiphospholipid antibody (APA) levels were tested by enzyme linked immunosorbent assay (ELISA). Plasma lipid levels were measured by routine methods.</p><p><b>RESULTS</b>No statistically significant differences were found in frequencies of genotypes and alleles of G1025C (Try316Ser) polymorphism between the controls and stroke patients. The serum levels of TG in the GC genotype of cerebral infarction patients and controls were markedly higher than those in GG genotype.</p><p><b>CONCLUSION</b>There was no association betweenG1025C (Try316Ser) polymorphism and stroke in Changsha Hans. G1025C (Try316Ser) polymorphism was associated with plasma lipid metabolism in Changsha Hans.</p>