[Preoperative planning of constitutional varus alignment in patients with varus knees and coronal femoral lateral bowing].

Objective: To investigate the preoperative constitutional varus alignment (CA) inpatients withvarus osteoarthritic knees and coronal femoral lateral bowing (FLB) by comparing the femoral axis of the patients and volunteers' with straight femoral shaft (SFS) and healthy knees, which would be used to guide the selection of valgus correction angle(VCA) of distal femur in total knee anthroplasty (TKA). Methods: From January 2018 to December 2018, 45 nonconsecutive patients (90 knees) with varus osteoarthritic knees and obvious FLB (>5°) received primary TKA,and 50 limbs with healthy knees and SFS in 25 volunteers were studied at Xinqiao Hospital.The medial proximal femoral angle (MPFA) and anatomic lateral distal femoral angle (a-LDFA) were measured and compared between the two groups.The VCA formed by distal femoral mechanical axis (DMA) and distal femoral anatomic axis (DAA) in patients and the VCA based on neutral mechanical axis (NMA) in volunteers were also compared. The Pearson's correlation analysis was performed between the angle of bowing (α) and the VCA based on NMA in patients. The measurement data was compared with t test between the two groups. Results: The results showed that the MPFA and the a-LDFA were not significantly different between the volunteers with SFS and patients with FLB (MPFA:84.9°±3.7° vs 85.6°±1.8° and a-LDFA:81.7°±1.7° vs 81.2°±1.6°, t=1.26, 1.70, both P>0.05). The VCA based on NMA in volunteers with SFS was 5.5°±0.6°(4.5°-7.4°), the preoperative DAA-DMA angle was 5.3°±0.7°(4.3°-7.8°) in patients with FLB, there was no significant difference between the two groups (t=1.70, P=0.09). The angle of bowing (α) was 7.9°±2.9° (6°-16°), the VCA based on NMA was 8.4°±1.5°(5°-10°), there was a significantly positive correlation between VCA and α (R=0.607, P<0.01). Conclusion: There is no significantly proximal or distal femoral deformity in patients with varus osteoarthritic knee and FLB (>5°), the degree of the DMA-DAAangle based on the DMA doesn't change with the increasing angular deformity of the bowing, then the bowing would be reserved if the distal femur is cut based on DMA in TKA and the preoperative CA should be restored successfully.

Xpert MTB/RIF Assay for Pulmonary Tuberculosis and Rifampicin Resistance in Children: a Meta-Analysis.

BACKGROUND: The Xpert MTB/RIF assay has been recommended by WHO to replace conventional microscopy, culture, and drug resistance tests. It simultaneously detects both Mycobacterium tuberculosis infection (TB) and resistance to rifampicin (RIF) within two hours. The objective was to review the available research studies on the accuracy of the Xpert MTB/RIF assay for diagnosing pulmonary TB and RIF-resistance in children. METHODS: A comprehensive search of Pubmed and Embase was performed up to October 28, 2014. We identified published articles estimating the diagnostic accuracy of the Xpert MTB/RIF assay in children with or without HIV using culture or culture plus clinical TB as standard reference. QUADAS-2 tool was used to evaluate the quality of the studies. A summary estimation for sensitivity, specificity, diagnostic odds ratios (DOR), and the area under the summary ROC curve (AUC) was performed. Meta-analysis was used to establish the overall accuracy. RESULTS: 11 diagnostic studies with 3801 patients were included in the systematic review. The overall analysis revealed a moderate sensitivity and high specificity of 65% (95% CI: 61 - 69%) and 99% (95% CI: 98 - 99%), respectively, and a pooled diagnostic odds ratio of 164.09 (95% CI: 111.89 - 240.64). The AUC value was found to be 0.94. The pooled sensitivity and specificity for paediatric rifampicin resistance were 94.0% (95% CI: 80.0 - 93.0%) and 99.0% (95% CI: 95.0 - 98.0%), respectively. Hence, the Xpert MTB/RIF assay has good diagnostic and rifampicin performance for paediatric pulmonary tuberculosis. CONCLUSIONS: The Xpert MTB/RIF is sensitive and specific for diagnosing paediatric pulmonary TB. It is also effective in detecting rifamnicin resistance. It can, therefore, be used as an initial diagnostic tool.

Estimate of 2009 H1N1 influenza cases in Shenzhen--the biggest migratory city in China.

The 2009 novel H1N1 influenza pandemic had a significant impact on Shenzhen's population with 2063 laboratory-confirmed human H1N1 cases and five deaths being reported. We used parameters from two population-based surveys and the Shenzhen Influenza Surveillance System to estimate the total number of H1N1 influenza infections in Shenzhen in the 2009 pandemic. The attack rate of influenza-like illness (ILI) in family households was 11·2% (95% CI 9·4-13·0), with 80·2% (95% CI 77·8-82·5) seeking medical care. The ILI attack rate in workers was 38·1% (95% CI 34·3-41·7) with 72·5% (95% CI 66·9-78·0) seeking medical care. The average H1N1 positive rate in individuals reporting ILI and testing by polymerase chain reaction was 22·7%. A total of 611 000-768 000 people, or 4·7-5·9% of the Shenzhen population, are estimated to have experienced H1N1 influenza. The estimated total number of cases of H1N1 is likely to be 330 times greater than the number of laboratory-confirmed cases.

Influenza surveillance in Shenzhen, the largest migratory metropolitan city of China, 2006-2009.

Shenzhen is one of the largest migratory metropolitan cities in China. A standardized influenza surveillance system has been operating in Shenzhen for several years. The objectives of the present study were to describe the epidemiology of influenza in Shenzhen and to assess the impact of pandemic H1N1 on influenza activity. An average rate of 71 cases of influenza-like illness (ILI)/1000 consultations was reported, which was greater than the rate in the preceding 3 years. Laboratory surveillance showed that the annual proportion of specimens positive for influenza was 25·4% in 2009, representing a significant increase over the proportions of 5·4%, 11·6% and 12·2% in 2006, 2007 and 2008, respectively. A total of 414 ILI outbreaks were reported in 2009, which was a marked increase compared to the previous 3 years. Influenza activity reached a record high in Shenzhen in 2009. Seasonal A/H3N2 was the dominant strain during the summer and was gradually replaced by pandemic H1N1. A semi-annual cycle for influenza circulation began to appear due to the emergence of pandemic H1N1.

Transcriptional analysis of a major capsid protein gene from Spodoptera exigua ascovirus 5a.

The major capsid protein (mcp) gene of Spodoptera exigua ascovirus 5a (SeAV-5a) was confirmed by aphidicolin viral DNA replication inhibition analysis to be a late gene. The 5' and 3' ends of mcp gene transcripts have been mapped. Primer extension analyses indicated that transcription of the mcp gene initiates from a cytosine 25 nucleotides (nt) upstream of the translation start codon. Two independent approaches by 3' rapid amplification of cDNA ends (3' RACE) and oligo (dT) cellulose binding assay suggested that SeAV-5a mcp mRNA is polyadenylated. Analyses by 3' RACE also revealed that mcp transcripts terminate at a U, either at 26 or 38 nt downstream of the translation stop codon. The putative 5' transcription control region of the SeAV-5a mcp gene shares similarities with other ascoviruses and Chilo iridescent virus (CIV), containing a conserved TATA-box-like motif (TAATTAAA) and an ATTTGATCTT motif upstream of it. The 3' downstream regions of the mcp gene of all the ascoviruses examined and CIV can form a stem-loop structure, and the ends of the mcp gene transcripts of SeAV-5a are within the predicted stem-loop region. This suggests that the stem-loop structure of the mcp gene might be involved in transcription termination.

Molecular cloning and characterization of a putative nuclear DEAD box RNA helicase in the spruce budworm, Choristoneura fumiferana.

RNA helicases play important roles in cellular processes such as pre-mRNA splicing, rRNA processing, ribosomal biogenesis, and translation. A full-length DEAD box RNA helicase cDNA (CfrHlc113) was isolated from the spruce budworm, Choristoneura fumiferana. CfrHlc113 contained the eight functional motifs, which are highly conserved in the DEAD box RNA helicase family, and an arginine-serine-aspartate (RSD) domain at its N-terminal end. CfrHlc113 was highly homologous to Rattus norvegicus HEL117 and human prp5 genes, both of which are suggested to be involved in RNA splicing. The results of Northern and Western blotting showed that expression of the CfrHlc113 gene was low or undetectable in eggs, larvae, pupae, and adults. High levels of expression were, however, detected in the three in vitro cultured cell lines, CF-203, CF-124T, and CF-70, which were developed from the midgut, ovaries, and neonate larvae, respectively. Immunocytochemistry revealed that CfrHlc113 protein was present exclusively in the nuclei of these cell lines.

Deficiency of the cysteine protease cathepsin S impairs microvessel growth.

During angiogenesis, microvascular endothelial cells (ECs) secrete proteinases that permit penetration of the vascular basement membrane as well as the interstitial extracellular matrix. This study tested the hypothesis that cathepsin S (Cat S) contributes to angiogenesis. Treatment of cultured ECs with inflammatory cytokines or angiogenic factors stimulated the expression of Cat S, whereas inhibition of Cat S activity reduced microtubule formation by impairing cell invasion. ECs from Cat S-deficient mice showed reduced collagenolytic activity and impaired invasion of collagens type I and IV. Cat S-deficient mice displayed defective microvessel development during wound repair. This abnormal angiogenesis occurred despite normal vascular endothelial growth factor and basic fibroblast growth factor levels, implying an essential role for extracellular matrix degradation by Cat S during microvessel formation. These results demonstrate a novel function of endothelium-derived Cat S in angiogenesis.

Glycation cross-links inhibit matrix metalloproteinase-2 activation in vascular smooth muscle cells cultured on collagen lattice.

AIMS/HYPOTHESIS: Extracellular matrix glycation has been proposed to contribute to the arterial stiffness observed in aging and diabetes. We examined whether matrix protein glycation regulates the proleolytic process through the manipulation of matrix metalloproteinases (MMPs) activation, using collagen fibrils model. METHODS: Vascular smooth muscle cells were cultured on control or glycated collagen fibrils. Matrix metalloproteinase-2 activation and the production of tissue inhibitors of metalloproteinase (TIMPs) were measured in the conditioned medium by using gelatin zymography and immunoblotting. Membrane type 1 matrix metalloproteinase (MT1-MMP) expression was also measured in cell lysates. RESULTS: When smooth muscle cells were cultured on collagen fibrils, pro-MMP-2 processing to active form was observed in the conditioned medium in coincidence with the increased MT1-MMP expression and the suppressed TIMP-2 production. Culturing smooth muscle cells on glycated collagen fibrils inhibited MMP-2 activation and attenuated MT1-MMP expression without the alteration of TIMP-2 production compared with control fibrils, indicating the possible mechanism of the suppression of MT1-MMP expression for the inhibition of MMP-2 activation on glycated collagen fibrils. Inclusion of aminoguanidine, an inhibitor of cross-linking formation, during collagen glycation restored the MMP-2 activation, suggesting the role of cross-links on the inhibition of MMP-2 activation. CONCLUSION/INTERPRETATION: These observations suggest that glycation-induced cross-linking formation in interstitial collagen contributes to arterial stiffness in aging and diabetes through the manipulation of matrix metalloproteinase activation along with the reduction of the susceptibility to proteolytic enzymes.

Activation of MMP-2 by Clostridium difficile toxin B in bovine smooth muscle cells.

Matrix metalloproteinase-2 (MMP-2) plays critical roles in cell migration through the breakdown of the extracellular matrix. Cell movements require dynamic actin reorganization, which is controlled by Rho family GTPases. In order to examine the relation between MMP-2 regulation and actin reorganization, we used several inhibitors of Rho family GTPases. Treatment of smooth muscle cells with Clostridium difficile toxin B known to inactivate Rho family GTPases activated MMP-2. However, neither C3 transferase, a Rho inhibitor, nor Y-27632, a specific inhibitor of Rho-kinase, induced MMP-2 activation. Treatment with C3 transferase and Y-27632 caused morphological changes into the round and stellate shape, respectively, by inhibition of actin stress fiber formation. In addition, toxin B treatment induced expression and processing of MT1-MMP, a major activator of MMP-2. Taken together, we suggest the involvement of Rho family GTPases, although inhibition of neither Rho nor Rho-kinase is sufficient, in the activation of MMP-2 through expression and activation of MT1-MMP.

Characterization of a single-nucleocapsid nucleopolyhedrovirus of Thysanoplusia orichalcea L. (Lepidoptera: Noctuidae) from Indonesia.

A single-nucleocapsid nucleopolyhedrovirus (NPV) isolated from Thysanoplusia orichalcea L. (Lepidoptera:Noctuidae) (ThorNPV) in Indonesia has tetrahedral occlusion bodies (OBs) with a width of 1. 22 microm (range = 0.803-1.931 microm). The length of the virion with an envelope averaged 0.29 and 0.23 microm without an envelope. ThorNPV was propagated in Pseudoplusia includens (Walker) and its authenticity was confirmed by sequence analysis of the polyhedrin gene of the ThorNPV produced in T. orichalcea and P. includens. Polyhedrin amino acid sequence analysis revealed that ThorNPV belongs to Group II of baculoviruses and is closely related to Trichoplusia ni single nucleocapsid NPV, sharing 97.6% sequence identity. Infectivity of ThorNPV against third instar P. includens was low, with a LD(50) value of 65,636 OBs/larva. Electron microscopy of infected tissues showed many polyhedra without virions embedded, which might explain the low virulence against P. includens. Differences in virion occlusion rates between individual cells in the same tissue suggested that the inoculum consisted of at least two variants that differed in the gene(s) controlling virion occlusion. In a host range test using the LD(50) value to P. includens against Spodoptera exigua, S. frugiperda, S. eridania, Anticarsia gemmatalis, Helicoverpa zea, Trichoplusia ni, and P. includens, P. includens was the only species infected. The virus infected primarily the fat body, tracheal epithelium, and hypodermis. The genomic size of the ThorNPV is 135 kb.

Circular configuration of the genome of ascoviruses.

A circular configuration of genomic DNA was observed in ascoviruses isolated from two species of insects of the family Noctuidae [fall armyworm (Spodoptera frugiperda) and cotton bollworm (Helicoverpa zea)] using restriction endonuclease (REN) digestion, conventional gel electrophoresis, pulsed-field gel electrophoresis and Southern hybridization analysis. This circular configuration of ascovirus genomic DNA was established based on the difference between linear and circular DNA in the numbers of fragments resolved on agarose gel electrophoresis after single and double REN digestion. Genomic DNA of ascoviruses was found to be sheared after purification.

Polyhedrin sequence determines the tetrahedral shape of occlusion bodies in Thysanoplusia orichalcea single-nucleocapsid nucleopolyhedrovirus.

A nucleopolyhedrovirus (NPV) isolated from the looper Thysanoplusia orichalcea L. (Lepidoptera: Noctuidae) (ThorNPV) is occluded in a tetrahedral protein matrix. The ORF of the ThorNPV polyhedrin gene contains 738 nt which code for 246 amino acids of the putative polyhedrin protein with an estimated molecular mass of 28,778 Da. The promoter of this gene is similar in length to the promoter of Spodoptera frugiperda NPV (SfMNPV), with a 5 nt deletion before the start codon compared to those of other NPVs. When the polyhedrin gene of Autographa californica NPV (AcMNPV), whose occlusion bodies (OBs) are polyhedral, was replaced by the polyhedrin gene of ThorNPV, which produces tetrahedral OBs, tetrahedral polyhedra with properly occluded virions were produced. This work establishes the importance of the polyhedrin protein sequence in determining OB shape. Leucine at position 43 of ThorNPV polyhedrin was identified as responsible for the tetrahedral shape of ThorNPV OBs by PCR-based site-directed mutagenesis. Susceptibility to alkaline buffer of OBs formed by recombinant AcMNPV (RECAcV) carrying the polyhedrin gene of ThorNPV was slightly greater than that of native ThorNPV OBs. The LD50 of RECAcV for third-instar beet armyworm (Spodoptera exigua) was significantly lower than that of AcMNPV (253 and 31 OBs per larva, respectively).