Objective To investigate the efficacy of the combination of improved intramedullary VSD drainage and contained antibiotics bone graft to treat chronic tubular bones osteomyelitis.Methods From March,2011 to December,2013,our department have total of 40 patients with chronic tubular bones osteomyelitis.Twenty cases (group A) treat with one-stage osteomyelitis debridement cortical bone slotted,contained antibiotic bone and autologous bone implants and wound repair.Twenty cases (group B) treat with improved intramedullary VSD drainage 3-5 days temporarily after osteomyelitis debridement cortical bone slotted,then contained antibiotic bone and autologous bone implants and wound repair.A retrospective comparison of two groups of an average residence time of wound drainage postoperative,bone bed bacterial culture positive rate,average healing time,the average time of hospital stay,the average bone healing time,and recurrence rate of osteomyelitis.Statistical analysis with T test was used for above independent parametric.Results The two groups were followed-up for 6-24 months,independent samples t-test was used for two groups in the wound healing time,bone healing time,the drainage tube removal time and the length of hospital stay,in group A bone bed bacteria culture positive rate was 40％,group B was 5％,group A infection relapse has 2 cases,1 case was debridement cured,1 case was amputation,and the recurrence rate of 10％.Group B without infection recurrence,and the recurrence rate of 0％ ; The healing time and hospital stay of intramedullary drainage surgery patients (18.05 ± 2.74 d and 22.65 ± 2.80 d,respectively,in group B) was significantly less than one-stage surgery patients (24.10 ± 8.20 d and 28.10 ± 9.35 d,respectively,in group A),but the bone healing time and the drainage tube removal tine of two groups.There was no significant difference (P ＞ 0.05).Conclusion Contained antibiotic bone and autologous bone implants with wound healing therapy after osteomyelitis debridement cortical bone slotted with improvement VSD intramedullary drainage to treat patient with tubular bones osteomyelitis was more effective,it worthy of clinical spread.

To unveil genetic variations between the predominant soybean mosaic virus (SMV) strains in China and in the USA, as well as to reveal the potential relevance between the similarity of gene sequences and the virulence of the viruses, we isolated and sequenced the coat protein (CP) gene of Chinese SMV strain SC7 by RT-PCR and compared the SC7 sequence with those of SMV strains from the USA. Analysis is showed that the CP gene of SC7 was 795 nucleotides in length and encoded 265 in amino acids'. The CP gene of SC7 and those of the strains from the USA exhibited 4%-5% nucleotide diversity and 1%-2% diversity amino acids. The conserved amino-acid sequence associated with aphid spread in the USA strains was DAG, and corresponded to DAD in SC7. The virulence of SC7 was greater than that of the SMV strains from the USA. Nevertheless, no clear relationships between sequence similarity of the CP genes from different strains and their virulence on differential hosts were found.
Amino Acid Sequence ; Capsid Proteins ; genetics ; China ; Molecular Sequence Data ; Mosaic Viruses ; Soybeans ; virology ; United States

AlM:To find better ways of treating ocular alkali burn, and to reduce the suffering of patients and social burden.METHODS:Totally 100 patients were graded according to the degree of chemical burns to four major groups, each half were randomly divided into the control group and the treatment group. Control group underwent conventional treatment. ln addition to conventional therapy, patients in each treatment group were also added a Breviscapine intravenous injection of 40mg daily. Corneal recovery time, changes in vision, degree of corneal opacity, number of corneal neovascularization and other complications were observed. Curative effects were analyzed statistically.
RESULTS:There was no significant difference in levelⅠgroup between control group and treatment group ( P>0. 05); There were significantly different in level Ⅱ, Ⅲand Ⅳ group ( P<0. 05 ). Compared to the degree of corneal opacity and the number of corneal neovascularization, the treatment group was obviously better than the control group(P<0. 05).
CONCLUSlON: Breviscapine in the treatment of ocular alkali burns can shorten the course of treatment, reduce corneal scarring, and improve vision.

Objective To construct cDNA entry library and cDNA expression library of Armillifer agkistrodontis (A.) nymphs and make a preliminary immunoscreening for the cDNA expression library.Methods The nymphs were collected from the Kunming mice infected experimentally with A.agkistrodontis eggs and the total RNA were extracted from the nymphs using TRIzol Reagent.After purifying the mRNA,the synthesized cDNAs were cloned into the donor vector pDONR222 by BP reaction of Gateway technology and the recombinants were transformed into the DH10B cells by electroporation,the cDNA entry library was obtained.Next,the expression vector pDEST17 was ligated with entry clones by LR reaction,and the recombinants were transformed into the BL21 (DE3) cells.Hence,the cDNA expression library was constructed.Then,the expression library was immunoscreened with the mixed sera of mice infected with A.agkistrodontis,and the insertions of positive clones were sequenced.After that,the open reading frame(ORF) of positive slone sequence,the homology of the screened genes and their encoded proteins were analyzed by Finder and BLAST (basic local alignment search tool) program of National Center of Biotechnology Information(NCBI),and the discovered new genes were submitted into the GenBank.Besides,the physico-chemical properties,secondary structure and B cell epitopes of encoded proteins were also analyzed by bioinformatics software.Results The average titer and total clones of the cDNA entry library were 1.45 × 105 CFU/ml(colony-forming unit,CFU) and 1.74 × 106 CFU,respectively,and the range of fragment length of the inserted cDNA was between 0.2-4.0 kb,with an average of 1.4 kb.The total clones of cDNA expression library were 1.00 × 105 CFU,and the fragment length of the inserted cDNA was between 0.3-2.2 kb,with an average of 1.0 kb.Five positive clones,coded S1,S5,A1,D1 and F1,respectively,were obtained through preliminary immunoscreening.The sequence and homology of the five positive clones were sequenced and analyzed by BLAST program.No significant similarities were found in pentastomida species,which meant that they were all novel genes of A.agkistrodontis.The gene sequences were submitted to GenBank,with the accession number from JQ180451 to JQ180455.Also,results obtained by bioinformatics software showed that the predictive encoding proteins were all potential to be valuable recombinant diagnostic antigens.Conclusions The cDNA library of A.agkistrodontis nymphs is successfully constructed,and five new genes of A.agkistrodontis are discovered.The establishment of cDNA library and the discovery of the new genes will lay a foundation for further studying the gene functions and screening the immunodiagnostic antigens.

Adult ; Bone Screws ; Female ; Fracture Fixation ; Fractures, Bone ; surgery ; Humans ; Internal Fixators ; Male ; Middle Aged ; Patella ; injuries ; surgery

The difficulty and expense of seeing a doctor was attributed to the out-of-level diagnosis and treatment, and the out-of-level treatment was due to the weakening of the basic medical service capacity. Since the new medical reform,the state has invested a lot, which led to the weakening of the primary medical service capacity. Exploring the institutional root of the weakening of grass-roots medical service ability could help to find the realistic path to enhance the service capacity of grass-roots medical institutions. Enhancing the service capacity of primary medical institutions was the only way to implement graded medical treatment. The administration of medical institutions restricted the improvement of service capacity of primary medical institutions, and the root of administration was the existing medical and health system. Only by starting from the reform of the system and realizing the administration of primary medical institutions, the service capacity of primary medical institutions could be enhanced.

Antiviral Agents ; adverse effects ; therapeutic use ; Female ; Hepatitis C, Chronic ; complications ; drug therapy ; psychology ; Humans ; Interferon-alpha ; adverse effects ; therapeutic use ; Mental Disorders ; complications ; Middle Aged

AIMTo explore the differentially expressed proteins between hypobaric hypoxic delayed preconditioning (HHDP) and normal mouse hippocampus.

METHODSAfter the animal model of HHDP was constructed, hippocampal proteins were obtained by a series of abstraction with lysis solution containing high concentration urea. As soon as isoelectric focusing and SDS-PAGE was performed. The resolved proteins in the 2-DE gels were visualized by Coomassie blue R-250. The gels were scanned, and the images were processed with PDQuest software. Differential proteins were exactly excised from the gels, destained and digested with trypsin. The peptides were isolated and sent for MALDI-TOF-MS testing. Database searching was performed using peptide masses obtained from MALDI-TOF-MS.

RESULTSAverages of 481 +/- 38 and 477 +/- 21 protein spots were detected in control gels and preconditioning gels, respectively. 169 +/- 6 protein spots were matched between these two types of gels. Among the matched spots, while the quantities of 21 +/- 12 spots in control gels increased by above 2 times than that in preconditioning one, the quantities of 33 +/- 10 spots in preconditioning gels increased by the same times than that in control one. The correlation coefficient between these two patterns were 0.7748 +/- 0.0267. 12 spots in preconditioning gels significantly increased compared with the control (P < 0.05, n = 4). Among 12 spots excised from the gels, perfect peptide mass fingerprinting spectrums of 8 spots were acquired. The results showed that one protein was fructose biphosphate aldolase A. Three proteins matched nothing might be new proteins. The other four proteins just matched the partial sequences of the proteins of database were no coincidence to it's isoelectric point and molecular weight. So they might be homological proteins.

CONCLUSIONMany proteins, for example fructose biphosphate aldolase A, has been differentially expressed in hippocampus of mice during HHDP. This may be one of the molecule mechanisms of HHDP.

Adaptation, Physiological ; Animals ; Electrophoresis, Gel, Two-Dimensional ; Hippocampus ; metabolism ; Hypoxia ; metabolism ; Hypoxia, Brain ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Proteins ; isolation & purification ; Proteome ; analysis

OBJECTIVETo elucidate that diffusion weighted imaging (DWI) can be used to predict the injured regions of neonatal brain with hypoxic-ischemic encephalopathy (HIE) in the early phase of injury, and to measure the apparent diffusion coefficient (ADC) values in the multiple regions of the brain.

METHODThe participants in this study were twenty-six infants with HIE from neonatology ward hospitalized between July 2006 and July 2009. Nineteen patients had severe HIE, and seven had moderate HIE. DWI and conventional magnetic resonance imaging (MRI) were performed for each case within the first 72 hrs. The ADC values of eight regions of interest (ROIs) were measured in ten cases with severe HIE (ADC values group). ROIs included posterior limb of internal capsule (PLIC), ventrolateral thalami, basal ganglia, perirolandic cortex, occipital cortex, centrum semiovale, brainstem, and frontal white matter. Twelve neonates were enrolled as the control subjects.

RESULTSDuring the first 72 hrs, the conventional MRI of 26 patients showed subarachnoid hemorrhage in 5, subdural hemorrhage in 2, and mild high signal intensity in the cortex of only one patient. In the 19 cases with severe HIE, abnormal signal intensities were seen in ventrolateral thalami and perirolandic cortex of 17 patients (89%), and the remaining 2 infants showed abnormal cortex and subcortical white matter. In 7 cases with moderate HIE, 4 had abnormal signal intensity in the cortex and subcortical white matter, 2 had abnormal periventricular white matter, and only one showed abnormal signal intensity in the ventrolateral thalami and perirolandic cortex. In the ADC values group, the average ADC values of posterior limb of internal capsule (PLIC), ventrolateral thalami, basal ganglia, perirolandic cortex, occipital cortex, centrum semiovale, brainstem, and frontal white matter respectively were 0.68 (0.56 - 0.88), 0.73 ± 0.13, 0.67 ± 0.11, 0.78 ± 0.22, 0.90 ± 0.16, 0.87 ± 0.21, 0.73 ± 0.19, 1.32 ± 0.22 × 10(-3) mm(2)/S. In the control group, the average ADC values of posterior limb of internal capsule (PLIC), ventrolateral thalami, basal ganglia, perirolandic cortex, occipital cortex, centrum semiovale, brainstem, and frontal white matter respectively were 0.96 (0.95 - 1.02), 1.02 ± 0.90, 1.15 ± 0.99, 1.08 ± 0.07, 1.09 ± 0.08, 1.39 ± 0.20, 0.96 ± 0.05, 1.58 ± 0.18× 10(-3) mm(2)/S. There was statistically significant difference in the average ADC values between each of 8 ROIs of infants with HIE and healthy neonates (P < 0.01).

CONCLUSIONIn the first days after birth, the major injured regions of severe HIE were ventrolateral thalami and perirolandic cortex, the minor injured regions were cortex and subcortical white matter. Multiple regions of moderate HIE were injured, including cortex with subcortical white matter, periventricular white matter, and ventrolateral thalami with perirolandic cortex. The ADC values of the regions with abnormal signal intensity decreased, also some regions with the normal signal intensity.

Brain ; pathology ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Hypoxia-Ischemia, Brain ; diagnosis ; Infant, Newborn ; Male

Human Zona Pellucida(ZP), which is a complex matrix surrounding oocytes,is comprised of three immunologically distinct glycoproteins(hZP1, hZP2 and hZP3). Because hZP3 possesses the sperm receptor activity and the acrosome-inducing activity, it has long been used as a candidate antigen to develop an immunocontraceptive vaccine. However, a large amount of native hZP3 protein is unavailable. It is an effective way to express hZP3 protein directly in vitro. Nevertheless, it had been reported that the rhZP3 protein produced in Pichia pastoris was not secreted but accumulated in the cells and could only be purified after being solubilized by strong denaturants. More unfortunately, after purification the final product required 6mol/L urea to maintain solubility. An improved project was advanced with the aim to express secreted and soluble rhZP3 protein in yeast. In this study, the fragment of hZP3 cDNA coding for aa 23 - 408, which the N-terminal leader was removed and most of the C-terminal transmembrane-like domain was reserved, was amplified by two PCR primers including EcoR I and Not I sites respectively and a His6 codon cassette was added to 5'-terminal. The hZP3 insert was incorporated into expression vector pPIC9K. The resulting recombinant yeast expression vector was designated pPIC9K-rhZP3. Linearized pPIC9K-rhZP3 was transformed into Pichia pastoris. After G418 selection, the recombinant Pichia pastoris strains were identified by PCR and the rhZP3 was expressed following the manufacturer' s protocol. Following induction with methanol, the rhZP3 protein was secreted and dissolved into the culture supernatant. SDS-PAGE and Western blot analyses showed that the apparent molecular weight of the expressed rhPZ3 proteins in yeast was smaller and a little size heterogeneity than native ones; after purified with Ni-chelating affinity chromatography, the final product's apparent molecular weight was about 32 - 34KD and their yield more than 20mg/L. We supposed that the C-terminal transmembrane-like domain be useful for secretion of rhZP3 into the culture supernatant and the expressed rhZP3 protein be incompletely digested by proteinases of Pichia into shorter fragments which all were glycosylated inhomogeneously. Fortunately, the fragments of rhZP3 protein can be recognized in Western blot by the polyclonal antibodies to porcine ZP3 which has showed a cross-reactivity with human ZP in vitro. It will be expected that the rhZP3 protein expressed in Pichia pastoris not only has immunogencity, say, it can rise antibodies in vivo to prevent spermatozoa-ovum binding, but also does not contain ovarian factors that might be the cause of undesired side effects, e.g. ovaritis and can be used as a safe immunogen in human antifertility vaccine research.
Blotting, Western ; Chromatography, Affinity ; Egg Proteins ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Genetic Vectors ; genetics ; Humans ; Membrane Glycoproteins ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; isolation & purification ; metabolism ; Zona Pellucida Glycoproteins