Flexor Hallucis Longus Transfer And V-Y Plasty: An Effective Treatment Modality for Chronic Achilles Rupture - A Case Series

@#Introduction: To assess outcomes of FHL transfer and V-Y plasty for chronic Achilles rupture due to insertional Achilles tendinopathy. Materials and methods: A case series of 12 patients was conducted between 1st January 2017 and 31st December 2018. The patients had short flexor hallucis longus tendon transfer with gastrocnemius lengthening by V-Y plasty for Achilles tendon rupture. Patients were allowed full weight bearing at six weeks post-operatively, and were followed up at three months and six months post-operatively, when the range of motion of the ankle was examined, and the outcome was assessed using the EFAS score. Results: Of the 12 patients in the study, the majority were males; the mean age was 50.6±8.96 years. A significant improvement in dorsiflexion and plantarflexion was noted at the six-month follow-up compared to the three-month follow-up (P=<0.001 for both). When compared to the normal side, dorsiflexion and plantarflexion of the affected ankle were significantly less at three months but were comparable at six months post-operatively. A significant improvement was noted in the mean EFAS score at the sixmonth follow-up (25.5±5.71) compared to three months (18.6±0.90) post-surgery (P=0.001). Males were also noted to have significantly higher EFAS scores at their six-month follow-up than females (P=0.022). In contrast, a negative correlation was noted between the European Foot and Ankle Society (EFAS) score at the final follow-up and age (P=0.011). Conclusion: FHL tendon transfer with V-Y plasty in chronic Achilles rupture due to insertional Achilles tendinopathy is an effective procedure resulting in the restoration of the ankle range of motion and improvement in functional scores.

Comparison of efficacy of platelet-rich plasma and extracorporeal shock wave for the treatment of chronic insertional Achilles tendinopathy / 中国骨伤

OBJECTIVE@#To compare clinical efficacy of platelet-rich plasma (PRP) and extracorporeal shock wave in treating chronic insertional Achilles tendinopathy.@*METHODS@#From February 2019 to August 2021, 42 patients with chronic insertional Achilles tendinopathy were selected and divided into PRP group(20 patients, 28 feet) and shock wave group (22 patients, 29 feet). In PRP group, there were 12 males and 8 females, aged 47.00(28.00, 50.75) years old, and the courses of disease ranged 7.00(6.00, 7.00) months;PRP injection was performed in the Achilles tendon stop area of the affected side. In shock wave group, there were 16 males and 6 females, aged 42.00(35.75, 47.25) years old;and the courses of disease was 7.00(6.00, 8.00) months;shock wave was performed in Achilles tendon stop area of the affected side and triceps surae area. Visual analogue scale (VAS) and Victorian Institute of Sport Assessment-Achilles (VISA-A) were applied to evaluate clnical effect before treatment, 1, 3 and 6 months after treatment, and satisfaction of patients was investigated.@*RESULTS@#VAS and VISA-A score in both groups were significantly improved at 1, 3 and 6 months after treatment than before treatment (P<0.05), VAS and VISA-A score in PRP group at 6 months after treatment were significantly higher than those at 1 and 3 months after treatment, and VAS and VISA-A score in shock wave group were lower than those at 1 and 3 months after treatment (P<0.05). There was no significant difference in VAS and VISA-A score between two groups before treatment, 1 and 3 months after treatment(P>0.05), while VAS and VISA-A score in PRP group were better than those in shock wave group at 6 months after treatment(P<0.05), and the satisfaction survey in PRP group was better than that in shock wave group(P<0.05).@*CONCLUSION@#PRP injection has a good clinical effect on chronic insertional Achilles tendinopathy with high patient satisfaction, and medium-and long-term effect of PRP injection for the treatment of chronic insertional Achilles tendinopathy is better than that of extracorporeal divergent shock wave.

Vitamin D-dependent rickets (VDDR) type 1: case series of two siblings with a CYP27B1 mutation and review of the literature / Raquitismo dependente de vitamina D: tipo 1: série de casos de dois irmãos com mutação CYP27B1 e revisão da literatura

Abstract Two siblings presented with clinical and biochemical features of rickets, initially suspected as hypophosphatemic rickets. There was no improvement initially, hence the siblings were reinvestigated and later diagnosed as having vitamin D-dependent rickets (VDDR) type 1 due to a rare mutation in the CYP27B1 gene encoding the 1α-hydroxylase enzyme. Both siblings improved with calcitriol supplementation. The initial presentation of VDDR is often confusing and algorithmic evaluation helps in diagnosis. We also present a brief review of the literature, including genetics.

Resumo Dois irmãos apresentaram características clínicas e bioquímicas do raquitismo, com suspeita clínica inicial de raquitismo hipofosfatêmico. Não houve melhora no início, portanto os irmãos foram reavaliados e, posteriormente, diagnosticados com raquitismo dependente de vitamina D (VDDR) tipo 1 devido a uma rara mutação no gene CYP27B1, que codifica a enzima 1a-hidroxilase. Ambos os irmãos melhoraram com a suplementação de calcitriol. A apresentação inicial do VDDR geralmente é confusa e a avaliação algorítmica ajuda no diagnóstico. Também apresentamos uma breve revisão da literatura, incluindo genética.

Achilles Tendon Lesions - Part 1: Tendinopathies / Lesões do Aquiles - Parte 1: Tendinopatias

Abstract Calcaneal tendon injuries are extremely common in the general population and in orthopedics routine care. Its increasing incidence, which is motivated by an aging population, improved access to the health care system, increased prescription of continuous-use medication, erratic participation in sports and other factors, has had a direct impact on society. Consolidated treatment options for tendinopathies lack quality scientific support for many modalities. New therapies have emerged to enhance nonsurgical approach outcomes and to reduce the number of patients requiring surgery. Although these operative procedures provide good pain relief and functional outcomes, they are costly and may lead to complications.

Resumo As afecções que acometem o tendão calcâneo são extremamente comuns na população geral e no cotidiano da atenção ortopédica. Sua crescente incidência, motivada pelo envelhecimento da população, pela melhora no acesso à saúde, pelo aumento na utilização de drogas de uso contínuo, pela errática participação esportiva e outros fatores, tem causado impacto direto na sociedade. As tendinopatias, ainda que hoje respaldadas por tratamentos consolidados, carecem de suporte científico de qualidade para muitas das suas recomendações. Novas terapêuticas têm surgido com o objetivo de potencializar o resultado da abordagem não-operatória e diminuir a quantidade de pacientes que necessitam de uma cirurgia. Esses procedimentos operatórios apresentam uma boa resposta álgica e funcional dos pacientes, no entanto não são livres de complicações e dos altos custos que os cercam.

Normative Data of Optimally Placed Endotracheal Tube by Point-of-care Ultrasound in Neonates

Objective: To derive normative data of the distance betweenoptimally placed endotracheal tube tip and arch of aorta byultrasound in neonates across different weight and gestation.Design: Cross-sectional study.Setting: Tertiary care neonatal intensive care unit from April 2015to May 2016.Participants: All neonates requiring endotracheal intubation wereeligible for the study.Methods: During intubation, insertional length was determinedusing weight-based formula. The distance between endotrachealtube tip and arch of aorta was measured by ultrasound.Endotracheal tube position was confirmed by chest radiograph.Results: Out of 133 enrolled infants, 101 (75.9%) had optimallyplaced endotracheal tubes. The mean (SD) distance betweenendotracheal tube tip and arch of aorta by ultrasound was 0.78(0.21) cm in infants <1500 g and 1.04 (0.32) cm in infants ≥1500 g.The regression equation to estimate insertional length fromweight, crown heel length (CHL), occipito-frontal circumference(OFC), nasal tragus length (NTL) and sternal length (SL) wereWt(kg)+4.95, 0.15×CHL(cm)+0.57, 0.22×OFC(cm)+0.49, 0.82×NTL(cm)+1.24 and 0.75×SL(cm)+2.26, respectively.Conclusion: Our study reports normative data of the distancebetween optimally placed endotracheal tube tip and arch of aortaby ultrasound in neonates. The distance between endotrachealtube tip and arch of aorta increases with increase in weight andgestation. Insertional length correlates strongly with all theanthropometric parameters

Literature Analysis of the Selection of Tyrosine Kinase Inhibitors for Chronic Myeloid Leukemia Patients with BCR-ABL35INS Mutation / 中国药房

OBJECTIVE： To provide reference for reasonable selection of tyrosine kinase inhibitors （TKI） in chronic myeloid leukemia （CML） patients with BCR-ABL35INS mutation. METHODS： Using “BCR-ABL insertional mutation” “ABL1 35ins mutation” “BCR-ABL c.1423_1424ins35” “ABL1 p.C475Tyrfs*11” as keywords， retrieved from CNKI， Wanfang database， Medline and COSMIC database， BCR-ABL35INS mutation CML patients were summarized and analyzed in respects of general information and treatment （treatment plan， patient compliance and drug withdrawal）， therapeutic effect （molecular biological mitigation and disease progress） and safety data （ADR） during 2007-2018. RESULTS： Totally 9 related literatures were included， involving 70 patients with BCR-ABL35INS mutation， all of them were foreign cases. Among them， 39 cases were male and 31 cases were female， with a median age of 49.2 years. The median time from the diagnosis of CML to the detection of BCR-ABL35INS mutation was 19 months. After detecting gene mutation， 39 cases were treated with imatinib （initial dose of 400 mg， po， once a day）， and molecular biological remission was achieved in 5 patients （12.9％）； 15 cases （38.5％） had molecular biological response but had disease progression； 8 patients （20.5％） had no response. Seventeen patients were treated with dasatinib （100 mg， po， once a day or 2 divided dose）， and 8 cases （47.1％） achieved molecular biological response. Twenty-one patients were treated with nilotinib （400 mg， po， 2 divided dose）， and 3 patients （14.3％） achieved molecular biological response； 2 patients achieved molecular biological response， but the disease progressed. Seven， three and seven of these patients stopped taking drugs due to adverse reactions， accounting for 17.9％， 17.6％ and 33.3％ respectively. All the ADRs were classified as grade 3-4 of the National Cancer Institute’s Common Terminology Criteria for Adverse Events， and most of them were hematological toxicity. CONCLUSIONS： CML patients with BCR-ABL35INS mutation are less likely to achieve molecular response on imatinib therapy but are more sensitive to dashatinib. In the course of treatment， we should strengthen the monitoring of blood system and other related indicators to ensure the safety and effectiveness of drug use.

Meta-analysis of insertion/deletion genetic variation of ACE gene and onset riskof type 2 diabetic nephropathy in Chinese population / 重庆医学

Objective To systematically assess the relation between angiotensin-I converting enzyme(ACE) gene insertion/deletion (I/D) variation and type 2 diabetic nephropathy (T2DN) onset risk among Chinese population.Methods The related literatures were retrieved from the China National Knowledge Infrastructure (CNKI) and Wanfang Data until June 1st,2016.The RevMan 5.0 was used to conduct the statistical analysis.The merge OR value and corresponding 95% confidence interval(95%CI) were used to assess ACE gene I/D polymorphism and T2DN onset risk.Results Totally 29 papers with 4 357 subjects were included according to the inclusion and exclusion standard,including 2 208 cases of DN and 2 149 cases of T2DM without DN.Meta analysis showed that compared with ACE gene I/D polymorphism I allele,D allele could significantly increase the risk of T2DM patients suffering from DN,the OR value and corresponding 95%CI were 1.44(1.25,1.66);the gene analysis showed that ACE gene I/D polymorphism loci were significantly correlated with DN onset risk in the Asian population.The corresponding relative onset risk OR and 95%CI were 1.42(1.15,1.76) and 1.75(1.46,2.10) in the dominant and recessive genetic model.The Begg′s test showed that the included data had no obvious publication bias existence.Conclusion ACE gene I/D polymorphism is closely correlated with the onset risk of T2DN,and D allele might be a risk genetic factor for DN occurrence in the patients with T2DM.

Significance of the Pars Interarticularis in the Cortical Bone Trajectory Screw Technique: An In Vivo Insertional Torque Study

STUDY DESIGN: Retrospective study. PURPOSE: Cortical bone trajectory (CBT), a more medial-to-lateral and shorter path than the traditional one for spinal fusion, is thought to be effective for severely degenerated vertebrae because screws are primarily stabilized at the posterior elements. We evaluated the efficacy of this approach through in vivo insertional torque measurement. OVERVIEW OF LITERATURE: There has been only one prior in vivo study on CBT insertional torque. METHODS: Between January 2013 and April 2014, a total of 22 patients underwent posterior lumbar fusion using the CBT technique. The maximum insertional torque, which covers the radial strength needed for insertion, was measured for 113 screws, 8 of which were inserted for L5 spondylolysis. The insertional torque for cases with (n=8) and without (n=31) spondylolysis of L5 were compared using one-way analysis of variance (ANOVA). To evaluate vertebral degeneration, we classified 53 vertebrae without spondylolysis by lumbar radiography using semiquantitative methods; the insertional torque for the 105 screws used was compared on the basis of this classification. Additionally, differences in insertional torque among cases grouped by age, sex, and lumbar level were evaluated for these 105 screws using ANOVA and the Tukey test. RESULTS: The mean insertional torque was significantly lower for patients with spondylolysis than for those without spondylolysis (4.25 vs. 8.24 in-lb). There were no statistical differences in insertional torque according to vertebral grading or level. The only significant difference in insertional torque between age and sex groups was in men <75 years and women ≥75 years (10 vs. 5.5 in-lb). CONCLUSIONS: Although CBT should be used with great caution in patient with lysis who are ≥75 years, it is well suited for dealing with severely degenerated vertebrae because the pars interarticularis plays a very important role in the implementation of this technique.

The Size of the Rotator Cuff and the Positional Relationship of Bony Landmarks: An Anatomical Study / 체질인류학회지

For treatment of the rotator cuff, locating the structure and position of the rotator cuff is crucial. The aim of this study is to identify the size of each rotator cuff and the locational relationship with bony landmarks, and to provide superficial landmarks for locating the tendon from the surface. Fifty-two shoulders from 26 cadavers were dissected and measured in a supine position. The central point was set as the most protrusive point on the greater tubercle of the humerus. The measurement of angles was described ventral as positive (+) and dorsal as negative (-) placing the long axis of the humeral shaft at 0degrees. The range of the angle which each rotator cuff tendon is attached to the humerus head was: 53.8~103.3 degrees for the subscapularis, -17.1~25.7 degrees for the supraspinatus, -68.4~-1.9 degrees for the infraspinatus, and -117.1~-75.7 degrees for the teres minor. The vertical inferior point drawn from the anterior edge of the acromion to the humerus was 7.5+/-11.7 degrees from the central point. The average position of the point was the midpoint of insertion of the supraspinatus tendon. The lateral horizontal point drawn from the acromial angle to the humerus was -49.4+/-14.3 degrees away and located at an average of 30% inferior to the infraspinatus tendon. Also the lateral horizontal point drawn from the most protrusive point of the coracoid process to the humerus was 63.1+/-14.7 degrees away and located at an average of 20% superior to the subscapularis tendon. Lastly, the most protrusive point of the lesser tubercle of the humerus was located at a range of 80.8+/-11.1 degrees and an average of 60% superior to the insertion of the subscapularis tendon. For the measurements of the size of the rotator cuff, there was no statistical difference between the left and right. However, the four measurements - the proximal width of the teres minor tendon, the proximal and distal width, and the length of the subscapularis tendon - showed statistically significant difference between the sexes (P<0.05). Therefore, to identify the location of the tendon structures by palpation for shoulder treatment, using the lesser tubercle for the subscapularis, the anterior edge of the acromion for the supraspinatus, and the acromial angle for the infraspinatus as landmarks is regarded to be effective.

Overexpression of kermit/dGIPC is associated with lethality in Drosophila melanogaster

Insertional mutagenesis is an important tool for functional genomics in Drosophila melanogaster. The insertion site in the KG00562 mutant fly line has been mapped to the CG8709 (herein named DmLpin) locus and to the 3’ of kermit (also called dGIPC). This mutant line presents a high lethality rate resulting from a gain of function. To obtain some insight into the biological role of the mutated locus, we have characterized the mutation and its relation to the high mortality of the KG00562 fly line. In this mutant, we did not detect one of the DmLpin transcripts, namely DmLpinK, but we did detect an unusual 2.3-kb mRNA (LpinK-w). Further investigation revealed that the LpinK-w transcript results from an aberrant splicing between the untranslated first exon of DmLpinK and the mini-white marker gene. Lack of DmLpinK or LpinK-w expression does not contribute to lethality, since heterozygous KG00562/Def7860 animals presented lethality rates comparable to those of the wild type. In contrast, the overexpression of kermit was associated with lethality of the KG00562 fly line. Significantly higher levels of kermit were detected in the Malpighian tubules of KG00562/+ flies that presented higher lethality rates than wild-type or KG00562/Def7860 animals, in which the lethality was rescued. In agreement with a recently reported study, our data support the hypothesis that misexpression of kermit/dGIPC could interfere with Drosophila development, with further investigations being needed in this direction.

Construction and identification of mutation of pgm gene attenuate the virulent of Brucella melitensis strain 16 / 中国地方病学杂志

Objective The construction of suicide plasmid vector could be used to make mutation of pgm gene which attenuates the virulent of Brucella melitensis strain 16, the research may lay a foundation for the development of novel live attenuated vaccines. Methods Sucrose sensitive gene as forward screening sign and fusion sequences of kanamycin resistance gene were constructed based on plasmid pucl9; pucS1.6K suicide plasmid vector was established by modifying pgm gene with fusion sequences of kanamycin resistance gene (insertion mutation); pgm gene mutation of Brucella melitensis strain 16 was obtained by electro transformation and mutation was confirmed by PCR amplification. Results The results showed that the identified Brucella melitensis strain 16 pgm gene was inactivated after insertion of kanamycin resistance gene, and the mutant pgm gene DNA fragment length was approximately 3525 bp, in line with expectations, Brucella pgm gene mutant melitensis strain 16 was successfully constructed. Conclusions The construction of suicide plasmid vector and precise mutation of Brucella melitensis strain 16 is successful, the study is not only provided an effective technology platform for constructing mutants of Brucella but also lays a foundation for the development of novel live attenuated vaccines.

Surgical Management of Insertional Achilles Tendinopathy with Haglund's Deformity: A Preliminary Report / 대한정형외과학회잡지

PURPOSE: We wanted to report on treatment outcomes in insertional Achilles tendinopathy patients with Haglund's deformity that were operated on using a central achilles tendon splitting approach. MATERIALS AND METHODS: We enrolled six cases of insertional tendinopathy with concomitant Haglund's disease who underwent an operation between January, 2007 and November, 2008 and for whom follow-up duration for more than 1 year were available. The mean age was 41.5 (21-62) years; the mean follow-up period was 16.3 (12-24) months. Resection of retrocalcaneal bony prominences and inflamed bursa with debridement of degenerative tendons was done using central splitting approach in all cases. A lateral ankle radiograph was used to assess the amount of bony resection. Clinical and functional evaluations were done before and after the operation. RESULTS: In all cases, retrocalcaneal bony prominences were completely resected. Clinical and functional outcomes were significantly improved. One case had a mild tingling distal to the operative lesion. There were no other complications. CONCLUSION: A central splitting approach is an efficient procedure that can provide sufficient exposure to remove pathologic lesions in patients with insertional Achilles tendinopathy accompanying Haglund's disease.

Novel PML-RARA Fusion Gene on Chromosome 17 in Acute Promyelocytic Leukemia with Normal Chromosome 15 and 17

We describe a patient with acute promyelocytic leukemia (APL) with no detectable cytogenetic abnormality of either chromosomes 15 or 17 who nevertheless had juxtaposition of promyelocytic leukemia (PML) and retinoic acid receptor-alpha (RARA) and expressed a chimeric transcript. Conventional cytogenetics showed the 46, XX. The metaphase fluorescence in situ hybridization (FISH) with a 5' PML and 3' RARA probe showed a juxtaposed PML-RARA fusion signal on one chromosome 17 homologue, an RARA signals on the other chromosome 17 homologue, and one PML signal on each chromosome 15 homologue. Our patient is presently in remission and doing well after chemotherapy with idarubicin and all trans retinoic acid (ATRA) treatment. Our results show that APL patients with cytogenetically normal chromosome 15 and 17 may, nevertheless, have involvement of both PML and RARA genes and as the prognostic outcome in APL is associated with the presence of a PML-RARA fusion, it is necessary to perform RT-PCR or FISH to aid diagnosis.

The Candida albicans AAA ATPase homologue of Saccharomyces cerevisiae Rix7p (YLL034c) is essential for proper morphology, biofilm formation and activity of secreted aspartyl proteinases

Proper morphology is essential for the ability of Candida albicans to switch between yeast and hyphae and thereby sustain its virulence. Here we identified, by differential screening, a novel C. albicans AAA ATPase encoding gene, CaYLL34 (RIX7), with enhanced expression in hyphae. Phylogenetic analysis suggests that CaYLL34 belongs to a [quot ]VCP-like[quot ] subgroup of AAA ATPases essential for yeast viability and contains a bipartite nuclear localization signal. Inactivation of one copy of CaYLL34, by the URA-Blaster method, generated the heterozygous mutant strain M61. This strain has severe phenotypic alterations, such as a highly increased vacuole, abnormal cell shape and reduced growth in different conditions. Also, major pathogenicity factors are affected in M61, for instance, a significant decrease of hypha formation (>90%), surface biofilm adhesion (86%) and secreted aspartyl proteinase activity (76.5%). Our results show that the partial impairment of CaYll34p cellular levels is sufficient to affect the proper cellular morphology and pathogenicity factors and suggest that this protein is required for biogenesis of ribosomal subunits. Accordingly, we propose that the product of CaYLL34 could be tested as a novel target for antifungal drugs.

Clinical Utility of Radiographic Measurements of Insertional Achilles Tendinitis with Haglund's Deformity / 대한족부족관절학회지

PURPOSE: It is to analyze the clinical utility of radiograhpic measurements for the insertional Achilles tendinitis patients who have Haglund's deformity and to evaluate the radiographic values related to Haglund's deformity in normal Korean population. MATERIALS AND METHODS: We used the angle of Fowler and Philip, parallel pitch lines (PPL) and Chauveaux-Liet (CL) angle as radiographic measurements. We examined 50 cases of insertional Achilles tendinits patients and another 50 cases of normal foot as a control. RESULTS: In normal feet, the mean value of angle of Fowler and Philip was 53.22 degrees, CL angle was -33.14 degrees and the PPL showed positive in 48%. In case of insertional Achilles tendinitis, the mean values were 55.39 degrees, -33.63 degrees, positive in 56% respectively. There were not statistically significant differences (p>0.05). CONCLUSION: The radiographic values for Haglund's deformity between insertional Achilles tendinitis feet and the normal feet did not show significant difference. Therefore, it seems that the clinical utility of radiographic measurements for the insertional Achilles tendinitis with Haglund's deformity is not useful and the development of new diagnostic methods as MRI and ultrasonography is required.

Clinical Utility of Radiographic Measurements of Insertional Achilles Tendinitis with Haglund's Deformity / 대한족부족관절학회지

PURPOSE: It is to analyze the clinical utility of radiograhpic measurements for the insertional Achilles tendinitis patients who have Haglund's deformity and to evaluate the radiographic values related to Haglund's deformity in normal Korean population. MATERIALS AND METHODS: We used the angle of Fowler and Philip, parallel pitch lines (PPL) and Chauveaux-Liet (CL) angle as radiographic measurements. We examined 50 cases of insertional Achilles tendinits patients and another 50 cases of normal foot as a control. RESULTS: In normal feet, the mean value of angle of Fowler and Philip was 53.22 degrees, CL angle was -33.14 degrees and the PPL showed positive in 48%. In case of insertional Achilles tendinitis, the mean values were 55.39 degrees, -33.63 degrees, positive in 56% respectively. There were not statistically significant differences (p>0.05). CONCLUSION: The radiographic values for Haglund's deformity between insertional Achilles tendinitis feet and the normal feet did not show significant difference. Therefore, it seems that the clinical utility of radiographic measurements for the insertional Achilles tendinitis with Haglund's deformity is not useful and the development of new diagnostic methods as MRI and ultrasonography is required.

Insertional Mutation of ftsH Gene in Streptococcus pneumoniae Causes Stress Sensitivities

FtsH is a membrane-bound, ATP-dependent protease involved in various cellular functions. To understand its roles in Streptococcus pneumoniae and host-pathogen interactions, we inactivated the ftsH gene of D39 strain by inserting a tetracycline-resistance (tet) gene. Several recombinants containing the tet cassette within the ftsH gene were confirmed by Western immunoblotting for the absence of pneumococcal FtsH protein that could cross-react with antiserum raised against Escherichia coli FtsH. Compared with the wild-type D39 strain, the ftsH null mutants grew slowly with encapsulation and alpha-hemolysis on blood agar plates, but failed to grow in liquid media other than Todd Hewitt yeast extract broth. Even fresh cultures of ftsH null mutants appeared gram-negative. When the incubation temperature of liquid cultures was shifted from 37degrees C to 40degrees C, the mutants gradually lysed, whereas the shift to 30degrees C abolished further growth. The mutants also exhibited increased sensitivity to salt and remarkable growth inhibition by optochin. These observations suggest that no functional FtsH protein in pneumococcal cells causes a loss of cell surface integrity, resulting in impairment of cell growth under normal and stressful conditions.

Insertional Mutation of ftsH Gene in Streptococcus pneumoniae Causes Stress Sensitivities

FtsH is a membrane-bound, ATP-dependent protease involved in various cellular functions. To understand its roles in Streptococcus pneumoniae and host-pathogen interactions, we inactivated the ftsH gene of D39 strain by inserting a tetracycline-resistance (tet) gene. Several recombinants containing the tet cassette within the ftsH gene were confirmed by Western immunoblotting for the absence of pneumococcal FtsH protein that could cross-react with antiserum raised against Escherichia coli FtsH. Compared with the wild-type D39 strain, the ftsH null mutants grew slowly with encapsulation and alpha-hemolysis on blood agar plates, but failed to grow in liquid media other than Todd Hewitt yeast extract broth. Even fresh cultures of ftsH null mutants appeared gram-negative. When the incubation temperature of liquid cultures was shifted from 37degrees C to 40degrees C, the mutants gradually lysed, whereas the shift to 30degrees C abolished further growth. The mutants also exhibited increased sensitivity to salt and remarkable growth inhibition by optochin. These observations suggest that no functional FtsH protein in pneumococcal cells causes a loss of cell surface integrity, resulting in impairment of cell growth under normal and stressful conditions.

Quantitative Measurement of Insertional Activity

Analysis of insertional activity is a routine part of the clinical electromyogrphic examination. It provides an information of muscle excitability but it's clinical significance has not perfectively accepted yet. This study was designed to evaluate clinical usefulness of insertional activity through quantitative analysis in the diagnostic field of pathology. Monopolar needle electrode was inserted briefly in the biceps brachii, paralumbar spinal and tibialis anterior muscles of the normal and denervated muscles. Total duration and spike duration of the insertional activity were measured 10 times in each muscle and averaged. Within spike duration we measured turns, mean amplitude, turns/amplitude, RMS, mean frequency and median frequency. The measured parameters of insertional activities were not significantly different according to the muscle in normal controls. In denervated muscles, the turns, mean amplitude, RMS, mean frequency and median frequency were decreased but turns/amplitude was increased compared to those of normal controls. But there were no difference in total duration and spike duration between normal and denervated muscles. In denervated muscles the muscle power was positively correlated with turns, mean amplitude, RMS, mean frequency and median frequency, and the grade of abnormal spontaneous activities was inversely correlated with turns, mean amplitude, RMS, mean frequency and median frequency. Therefore quantitative analysis of insertional activity could be a useful method for the diagnosis of neuromuscular disease.

Construction of human antisense BMP-2 retrovirus expression vectors and its biological effects on human osteosarcoma cells / 中华骨科杂志

Objective To probe into the mechanisms of bone morphogenetic protein-2 (BMP-2) in osteosarcoma and to provide basis for gene therapy. Methods A 1.0 kb cDNA fragment of human BMP-2 gene was inserted reversely into PDOR and Human antisense BMP-2 retrovirus expression vector was constructed. The recombinant retroviral vector was transfected into human osteosarcoma cells OS-9901 with liposome AMINE that expressed abundant BMP. Positive cell clones were selected with G 418. The expression of BMP and proliferating cell nuclear antigen (PCNA) was determined by immunohistochemical ABC methods. The osteosarcoma cell cycle was analysed by flowcytometry, and the changes were observed by electronmicroscope. Results The expression of cellular BMP and PCNA in the transfected osteosarcoma cells decreased obviously (t=24.01， 26.09, respectively, P