Objective To develop TaqMan real-time PCR assay for detection and identification of streptococcus pneumonia iso-lated from children CAP.Methods Based on the sequences of lytA gene,primers and probe were designed and the assay was optimized.Then 1 504 sputum samples were detected by culture and the developed assay with double-blind testing.Results The lower limit of detection in the developed real-time PCR assay was 18.75 cfu/PCR,and had no cross reaction.141 strep-tococcus pneumonia strains were detected from 1 504 samples and 140 were isolated by culture.The whole process just nee-ded 2.5 h.Conclusion The established assay is rapid,simple,high sensitivity and specificity.It is not only valuable for the i-dentification of streptococcus pneumonia,but also provide evidence for antibiotic therapy.

Objective To study the diagnostic value of pencilliosis marneffei (PM)in a non-HIV-infected child with the com-bined detection of aspergillosis galactomannan,fungus Glucan(1-3)-β-D and boold culuture.Methods The venous blood specimen from the child was collected for the quantified detection of aspergillosis galactomannan,fungus Glucan(1-3)-β-D. The growth and colonial morphology of fungus was inspected with the positive blood culture and the characteristics of fun-gus smear were observed under microscope.Results The result of aspergillosis galactomannan was 14.45 μg/L and fungus Glucan (1-3)-β-D 77.14 pg/ml.Penicillium marnrffei was identified using blood culture.It was mycelia form under 25℃ and the salouraud medium produced water soluble claret-red pigment produced.It was mycelia form under 35℃ and the colony was gyri creases,the characteristic broom-like hypha and separation hypha could be found under microscope.Conclusion It is effective for the early diagnosis and therapy of PM with the combination detection of aspergillosis galactomannan,fungus Glucan (1-3)-β-D and boold culuture and have better clinical diagnosis value.

Objective To develop SYBR Green I real-time PCR assay for detection and identification of Hepatitis B virus. Methods Based on the sequences of Hepatitis B virus gp1 gene,primers were designed.The reaction assay and thermal cyc-ling profile were optimized.The positive standard was from recombinant clone.Both the developed assay and Zhejiang kuake biotechnology company’s assay were applied in 100 patients serum.Results The detection limit was between 5×102 copies/ml to 5×108 copies/ml with a good liner correlation and no cross reaction.The whole process just needed 2.5 h.Comparing with the company products,the sensitivity and specificity of the developed assay were 100% and 92.5% respectively.Con-clusion The established assay is rapid,simple,high sensitivity and specificity.It is not only valuable for the identification of Hepatitis B virus patients,but also provide accurate quantitative analysis for HBV patients.

Early-onset facioscapulohumeral muscular dystrophy is a rare clinical syndrome characterized by severe muscle weakness started in early childhood,with extramuscular manifestations such as retinal vascular tortuosity,sensorineural hearing loss and epilepsy.Herein we report a case with early-onset facioscapulohumeral muscular dystrophy and Coats syndrome.Early diagnosis of Coats syndrome is critical for the prognosis.

Objective: To explore the application value of protective device in microwave ablation at canine liver risk area, and the role of the device in reducing complications during ablation. Methods: Six healthy mongrel dogs were randomly divided into two groups: group A, used protective devices; group B, unprotected. Conventional gray-scale ultrasound and contrast-enhanced ultrasound (CEUS) were performed before treatment to identify the ablation area (the right lobe of the liver near diaphragm, 1 cm from the surface of the liver). The two groups were treated percutaneous puncture liver microwave ablation under real time ultrasound-guided with the same ablation power (50 W) and equivalent ablation time (600 s). Liver specimens were evaluated histological examination to evaluate the necrotic area of ablation and the degree of diaphragm′s injury. Results: ①There was no significance difference in the ablation range between two groups[ (3.3±0.1)cm vs (3.5±0.1)cm, P=0.184]; ②The tolerance of group A was better than group B during ablation treatment; ③The mean maximum ablation diameter in group A was significantly lower than that in group B [(2.1±0.1)cm vs (5.3±0.2)cm, P=0.001]. Meanwhile the pathological results showed that the damage degree in group A was significantly lower than that in group B. Conclusions This protective device has an obvious protective effect for microwave ablation of the risk area in liver (adjacent diaphragm, hepatic envelope). Our experiment suggests that this protective can reduce the damage of adjacent organ and the complications during the ablation surgery.

OBJECTIVE: To explore the clinical characteristics and genetic variant of a patient with desminopathy manifesting with atypical symptoms. METHODS: A patient who was admitted to the Department of Neurology of Jing'an District Central Hospital on February 24, 2021 was selected as the study subject. Clinical data, laboratory tests, muscle pathology, muscle magnetic resonance imaging (MRI) and genetic testing of the patient were retrospectively analyzed. RESULTS: The patient had developed myalgia after lower limb activity, and gradually developed asymmetrical muscle weakness and atrophy of the lower limbs. Cardiac examination revealed atrioventricular block and decreased left ventricular diastolic function. Muscle MRI showed that semitendinosus, sartorius, gracilis, fibula, gastronemius and supinator muscles were selectively involved at the early stage. Muscle biopsy confirmed pathological changes of desmin positive myofibrils. Genetic testing revealed that the patient has harbored a c.1024A>G (p.n342d) missense variant in exon 6 of the DES gene. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was rated as likely pathogenic (PS4_moderate+PM2_supporting+PP3_moderate+PP1). CONCLUSION Desmin disease has a great clinical heterogeneity. Postexercise myalgia of lower limbs is a rare clinical phenotype. For patients harboring the c.1024A>G (p.n342d) variant of the DES gene, in addition to semitendinosus and fibula, Cardiac involvement is relatively insidious and easy to be ignored in clinic. Timely muscle MRI, muscle biopsy and gene detection will help the early diagnosis of the disease.
Humans ; Myalgia/genetics* ; Desmin/genetics* ; Retrospective Studies ; Muscle, Skeletal ; Lower Extremity ; Mutation