Animals ; Animals, Newborn ; Cell Hypoxia ; Cell Lineage ; Cell Proliferation ; Cells, Cultured ; Oligodendroglia ; cytology ; Rats ; Rats, Sprague-Dawley

Objective To explore the effective method of treating childhood acute aplastic anemia(AAA) by overviewing the curative effect of therapeutic alliance to the children diagnosed as AAA with immunodepressants.Methods Retrospective analysis was performed on 13 patients diagnosed as AAA,who were treated by high dose of methyllprednisolone,antilymphocyte globulin,ciclosporin A,large dose of gamma globulin,granulocyte colony-stimulating factor and blood transfusion in 4 years.Results Among 13 cases,9 cases were healing well on the whole,one was relieved,one was improving,two were ineffective(one of them was dead).In 9 cases who were healing,WBC took(24.60?8.86) days to reach 4?10~9/L,hematoglobin took(152.22?68.88) days to reach normal and platelet took(125.55?55.86) days to reach 80?10~9/L.During the procedure of treating,infection appeared in 7 cases,the ALT and AST was high in 2 cases,hypoproteinemia and edema in 1 case,gross hematuria in 2 cases,gingival hyperplasia obviously and dedentition in 1 case.Conclusion Therapeutic alliance with immunodepressants to childhood AAA is a safe and effective method.

OBJECTIVETo study the proliferation effect of the AM supernatant incubated activation of p38 mitogen activated protein kinases (p38MAPK) signal transduction pathway in human embryonic lung fibroblasts, and to participate in the development of fibrosis in silicosis.

METHODSThe silicotic alveolar macrophages were collected by bronchoalveolar lavage and incubated in vitro in the DMEM medium containing SiO₂ (50 µg/ml) and DMEM medium without SiO₂ for 18 h. Then the AM supernatant incubated for 18 h was collected. HELFs were isolated by organize paste block method, and incubated with AM supernatants. HELFs were divided into four groups: blank control groups, AM groups, SiO₂ + AM groups, SB203580 + SiO₂ + AM groups. The proliferation in the HELF was detected with MTT method and Flow cytometry.

RESULTSThe proliferation in the HELF acted with the conditioned AM supernatant fluid were more than blank control groups, AM groups and SB203580 + SiO₂ + AM groups [average optical density: (0.48 ± 0.03) vs (0.29 ± 0.01), (0.38 ± 0.02), (0.33 ± 0.03)], the values with MTT method were statistically different (P < 0.05); Proliferous index with flow cytometry in SiO₂ + AM groups (18.12 ± 0.82) was bigger than blank control groups (9.24 ± 0.48), AM groups (14.76 ± 0.43) and SB203580 + SiO₂ + AM groups (11.71 ± 0.70) and the values were statistically different(P < 0.05).

CONCLUSIONSThe AM supernatant stimulated by silicon dioxide can accelerate the proliferation in the HELF by activation of p38MAPK signal transduction pathway.

Adult ; Cell Proliferation ; Cells, Cultured ; Culture Media, Conditioned ; Fibroblasts ; cytology ; drug effects ; pathology ; Humans ; Lung ; cytology ; MAP Kinase Signaling System ; Macrophages, Alveolar ; cytology ; Male ; Signal Transduction ; Silicon Dioxide ; pharmacology ; Silicosis ; metabolism ; pathology ; p38 Mitogen-Activated Protein Kinases ; metabolism

AIM: To investigate the effects of Rhodiola sachalinensis on ocular blood flow in diabetic retinopathy rats. METHODS:A total of 90 SD rats were randomly divided into control group (n=30), model group (n= 30) and intervention group (n=30). Rats in the model group and intervention group were fed with high glucose and high fat diet and injected with streptozotocin (40mg/kg) in order to construct DR model rats,while rats in the control group were fed with basic diet and injected with the same amount of normal saline. After 4wk, the rats in the intervention group were injected with Rhodiola sachalinensis injection(10mL,one per day),while rats in the control group and the model group were injected with normal saline. The course of intervention treatment was 4wk. The ocular blood flow in rats was detected by laser doppler flowmetry, the apoptosis of retinal cells in rats was detected by TUNEL method, the expression of vascular endothelial growth factor (VEGF), glial fibrillary acidic protein ( GFAP ) and glutamate/aspartate transporter (GLAST) in retina of rats was detected by RT-PCR method and Western blot method. RESULTS:The whole blood viscosity,whole blood high shear viscosity,low shear blood viscosity and erythrocyte sedimentation of rats in the model group and intervention group were higher than those in the control group (P<0 01),while the indexes of intervention group were lower than that of model group (P<0.01). The PSV and EDC of rats in the model group and intervention group were lower than those in the control group (P<0.01), and the Rl was higher than that in control group (P<0.01). The PSV and EDC in the intervention group were higher than those in the model group (P<0.01), and Rl was lower than that in the model group (P<0.01). The apoptosis rate of retinal cells of model group and intervention group was higher than that of the control group (P<0.01), that in the intervention group was lower than that in the model group (P<0.01). The expression of VEGF and GFAP in retina tissue of model group and intervention group was higher than that of the control group (P<0.01), and the expression of GLAST was lower than that in the control group(P<0 01). The expression of VEGF and GFAP in the intervention group was lower than that in the model group(P<0.01),and the expression of GLAST was higher than that in the model group (P<0.01). CONCLUSION: Rhodiola sachalinensis injection can significantly improve the ocular blood flow in diabetic retinopathy rats, reduce the apoptosis of retinal cells in rats, down regulate the expression of VEGF and GFAP, and up regulate the expression of GLAST.

This study aimed to investigate the reconstruction of the thumb and finger extension function in patients with middle and lower trunk root avulsion injuries of the brachial plexus.From April 2010 to January 2015,we enrolled in this study 4 patients diagnosed with middle and lower trunk root avulsion injuries of the brachial plexus via imaging tests,electrophysiological examinations,and clinical confirmation.Muscular branches of the radial nerve,which innervate the supinator in the forearm,were transposed to the posterior interosseous nerve to reconstruct the thumb and finger extension function.Electrophysiological findings and muscle strength of the extensor pollicis longus and extensor digitorum communis,as well as the distance between the thumb tip and index finger tip,were monitored.All patients were followed up for 24 to 30 months,with an average of 27.5 months.Motor unit potentials (MUP) of the extensor digitorum communis appeared at an average of 3.8 months,while MUP of the extensor pollicis longus appeared at an average of 7 months.Compound muscle action potential (CMAP) appeared at an average of 9 months in the extensor digitorum communis,and 12 months in the extensor pollicis longus.Furthermore,the muscle strength of the extensor pollicis longus and extensor digitorum communis both reached grade Ⅲ at 21 months.Lastly,the average distance between the thumb tip and index finger tip was 8.8 cm at 21 months.In conclusion,for patients with middle and lower trunk injuries of the brachial plexus,transposition of the muscular branches of the radial nerve innervating the supinator to the posterior interosseous nerve for the reconstruction of thumb and finger extension function is practicable and feasible.

Objective To explore the relationship between immune and pathogenesis of amyotrophic lateral sclerosis (ALS) in the investigation of neurofilaments phosphorylation and ultrastructure features in spinal cord ventral horn motor neuron injury mediated by immune.Methods Using transmission electron microscope,we studied the uhrastructure features of abnormal accumulations of neurofilaments (NF) in motoneuron of the spinal cord ventral horn,and immunohistochemically investigated neurofilaments phosphorylation.Results Electron microscope found that there was abnormal accumulation of interwoven NFs in motor neuronal perikarya and proximal axons.Immunohistochemical study revealed that the SMI-32 immunoreactive positive neurons (12.00?1.05),compared with control (18.00?1.83),were reduced (P

The clinical experiences and proven cases of distinguished doctor of TCM, professor WU Xu, on acupuncture for acute upper abdominal pain is introduced. Professor WU's manipulation characteristics of acupuncture for acute upper abdominal pain, including acute cholecystitis, kidney stone, acute stomach pain, are one-hand shape but both hands in nature, moving like Tai Chi, force on the tip of needle, movement of qi mainly. The main technique posture is one-hand holding needle with middle finger for pressing, the needle is hold by thumb and index finger, and is assisted by middle finger. The special acupuncture experience of emergency is treatment according to syndrome differentiation, combination of acupuncture and moxibustion, selecting acupoint based on experience, blood-letting acupuncture therapy and so on.
Abdominal Pain ; therapy ; Acupuncture Points ; Acupuncture Therapy ; Acute Pain ; therapy ; Adult ; Female ; Humans ; Male

OBJECTIVETo determine the difference of post-operative mortality between ORIF (open reduction internal fixation) and hip replacement for the treatment of intertrochanteric fracture in elderly by using survival analysis.

METHODSThe clinical data of 110 patients above 60 years old who underwent surgical treatment (ORIF or hip replacement) for the intertrochanteric fracture between April 2003 and May 2013 were retrospectively analyzed. Among the patients, 83 cases were treated with ORIF (ORIF group), there were 32 males and 51 females, aged from 61.44 to 98.75 years old with an average of (78.52 ± 7.98) years old; and 27 cases were treated with hip replacement (arthroplasty group), there were 8 males and 19 females, aged from 71.82 to 96.54 years old with an average of (79.99 ± 6.11) years old. A survival analysis was performed on the clinical data by using SPSS 110 software. The survival rate of 1-year,2-year, 5-year and the mean survival time for the total patients, the mortality rate of 1-year, 2-year in each group, the survival rate of 1-year, 2-year and mean survival time and survival curve in each group were included.

RESULTSAll wounds achieved primary healing and no deaths were found in stay hospital. All patients were followed up from 1 to 125 months with an average of (46.93 ± 29.53) months. Among all 110 cases, 31 were dead and 79 survived. The survival rate of 1-year, 2-year and 5-year was (90.7 ± 2.8)%, (82.5 ± 3.9)% and (57.6 ± 6.5)%, respectively,while the ensemble mean survival time was (84.137 ± 5.902) months. The mortality rate of 1-year, 2-year in ORIF group was 7.2% and 12.0%, respectively; and in arthroplasty group, there was 14.8% and 25.9%, respectively. There was no significant difference in mortality rate of 1-year and 2-year between two groups. According to the survival analysis of the ORIF group, the survival rate of 1-year, 2-year was (92.6 ± 2.9)%, and (85.8 ± 4.3)%, respectively, and the mean survival time was (87.508 ± 6.063) months. In arthroplasty group, the survival rate of 1-year, 2-year was (85.2 ± 6.8)% and (73.9 ± 8.5)%,and the mean survival time was (67.294 ± 11.180) months. There was significant difference in mean survival time between two groups (P < 0.05).

CONCLUSIONORIF can achieve a better postoperative survival compare with hip replacement in treating intertrochanteric fracture in elderly.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; Female ; Fracture Fixation, Internal ; Hip Fractures ; mortality ; surgery ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies

To combine the fibrinolytic with anticoagulant activities for therapy of thrombotic deseases, a fusion protein made of tissue-type plasminogen activator (t-PA) and hirudin was constructed and expressed in chia pastoris. To improve thrombolytic properties of t-PA and reduce bleeding side effect of hirudin, FXa-recognition sequence was introduced between t-PA and hirudin molecules.The anticoagulant activity of hirudin can be target-released through cleavage of FXa at thrombus site. t-PA gene and hirudin gene with FXa-recognition sequence at its 5'-terminal were obtained by RT-PCR and PCR respectively. The fusion protein gene was cloned into plasmid pIC9K and electroporated into the genome of Pichia pastoris GS115. The expression of fusion protein was induced by methanol in shaking flask and secreted into the culture medium. Two forms of the fusion protein, single-chain and double-chain linked by a disulfide bond (due to the cleveage of t-PA at Arg275-Ile276), were obtained. The intact fusion protein retained the fibrinolytic activity but lacked any anticoagulant activity. After cleavage by FXa, the fusion protein liberated intact free hirudin to exert its anticoagulant activity. So, the fusion protein is a bifunctional molecule having good prospect to develop into a new targeted therapeutic agent with reduced bleeding side effect for thrombotic diseases.
Animals ; Cloning, Molecular ; Electroporation ; Hirudins ; biosynthesis ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Plasminogen Activator ; biosynthesis ; genetics

BACKGROUNDRNA interference (RNAi) is a powerful tool to silence gene expression post-transcriptionally. Our previous study has demonstrated that small interfering RNAs (siRNAs) have sufficiently inhibited hepatitis B virus (HBV) replication and expression in vitro. In this study we observed the RNAi-mediated inhibitory effects on HBV replication in mice models and accessed the specificity of these effects.

METHODSA mutant RNAi vector (pSI-C mut) with two base pairs different from the original target gene sequence at the RNAi vector (pSI-C) was constructed according to the method described in this study. A mouse model of acute hepatitis B virus infection was established by injecting naked plasmid pHBV1.3 via the tail vein with acute circulatory overload. pSI-C, pSI-C mut and the irrelevant RNAi control plasmid for green fluorescent protein (GFP) gene, pSIGFP were respectively delivered with pHBV1.3 by tail vein injection method. Six days post injection, enzyme-linked immunosorbent assay (ELISA) assay was used to measure the concentration of HBV surface antigen (HBsAg) in mouse serum, immunohistochemical straining method was used to visualize the expression of HBV core protein (HBcAg) in liver tissues, and the transcriptional level of HBV C mRNA in liver tissues was detected by reverse transcriptase PCR (RT-PCR) analysis.

RESULTSInjection of pSI-C exerted magnificent and specific inhibitory effects on the replication and expression of HBV in the murine model. After 6-day post-injection (p.i.), the OD values were shown to be 5.07 +/- 1.07 in infecting group and 0.62 +/- 0.59 in pSI-C group. The concentration of HBsAg in pSI-C group was significantly lower than that in infecting group (P < 0.01). Liver intracellular synthesis of viral core protein was sharply reduced to 0.9% +/- 0.1%, compared with 5.4% +/- 1.2% of positive hepatocytes in infecting group (P < 0.01), and the transcriptional level of HBV C mRNA was greatly reduced by 84.7%. However, the irrelevant RNAi control plasmid (pSIGFP), and the pSI-C mut did not show the same robust inhibitory effects as pSI-C.

CONCLUSIONpSI-C exert efficient and specific inhibitory effects on HBV replication and expression in mice models.

Animals ; Hepatitis B ; therapy ; virology ; Hepatitis B Core Antigens ; biosynthesis ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; RNA Interference ; RNA, Messenger ; analysis ; RNA, Small Interfering ; therapeutic use ; RNA, Viral ; analysis ; Virus Replication