ObjectiveTo establish a novel rapid detection method based on PCR-single-strand-conformational polymorphism (PCR-SSCP) to determine mutation of streptomycin-resistance associated rpsL and rrs genes in isolates of Mycobacterium tuberculosis (MTB).MethodsStreptomycin-resistance of 112 MTB isolates was detected using the routine drug susceptibility test,and a special PCR-SSCP assay was established.The mutations of rpsL and rrs genes in streptomycin-resistant MTB isolates were detected by PCR-SSCP and PCR direct sequencing (PCR-DS) ; the results from two techniques were compared.Results All isolates had both rpsL and rrs genes.Fifty-two isolates (46.4％) were streptomycin susceptible,in which only 1 isolate showed abnormal PCR-SSCP fragments from rrs gene,and the specificity of PCR-SSCP was 98.1％ (51/52).Sixty isolates (53.6％) were streptomycin-resistant,in which 46 (76.6％) and 11 ( 18.3％ ) isolates presented the abnormal PCR-SSCP fragments of rpsL and rrs gene,respectively.One streptomycin-resistant isolate showed abnormal PCR-SSCP fragments from both rpsL and rrs genes.The sensitivity of PCR-SSCP was 93.3％ (56/60).ConclusionThe PCR-SSCP that established in this study is a specific and sensitive method for rapid detection of the streptomycin-resistance associated mutations in rpsL and rrs genes of MTB.

The effects of gradual tibia lengthening at different speeds on the histomorphology of tibial nerve were studied in 30 rabbits by self-made external distraction device. It was showed that the leg was tracted at the speed of 1 mm/d, the tibial nerve was lengthened correspondingly. Gradual stress could stimulate the growth of nervous tissue and no nervous functional and constructural injuries were found When the leg was distracted at the speed of 2 mm/d, limb elongation within 40% was safe, otherwise, severe nervous damage would occur.

ThestudiesonthemaincomponentsinEucommiaulmoidesfromdomesticandabroadweresummarizedinthepaper, which can provide the reference for the effective component study and quality control of the Chinese herb.

79 cases (180 amputated fingers) underwent gradual distraction through phanlanges. The bone elongation obtained varied from 1 to 3.2 cm, with an average of 2.2 cm. Finger stumps were examined by measurement of 2 point discrimination (2PD) and sensative nerve conduction velocity as well as SO, both before and post lengthening. We confirmed that it was safe to distract the finger stump at a daily rate of 1 mm. Several fingers can be lengthened simultaneously using our device. The adjacent joint was also stretched other than compressed. Therefore, the lateral accessory ligament contraction and joint crush injury were avoided. The long-term length increase was maintained by subperiosteal osteotomy. Gradual finger lengthening is a simple, safe and effective method.

ObjectiveTo investigate the value of MR diffusion tensor imaging (DTI) of optic nerve in the estimation of optic nerve changes of primary chronic angle-closure glaucoma (PCACG).Methods Twenty-five patients with PCACG including monocular involvement in 4 patients and binocular involvement in 21 patients and involving 46 eyes in which 24 right eyes and 22 1eft eyes,and 20 normal volunteers were enrolled.Conventional MRI and DTI were performed on all subjects using Magnetom Tim 3.0 T MRI.Fractional anisotropy( FA),mean diffusivity ( MD),axial diffusivities ( λ ∥ ) and radial diffusivities ( λ ⊥ )were measured and then compared between patients group and control group and between left eyes and right eyes.Two independent samples t-test and paired t-test were used.ResultsOn conventional MRI,thinner optic nerve with vaginal cavity widened slightly was found in 8 optic nerves of 6 patients.The value of FA,λ∥,λ⊥ and MD of 24 right optic nerves in patient group was(0.27 ± 0.09) × 10-3,(2.30 ±0.26) × 10 - 3,( 1.55 ± 0.35 ) × 10 - 3,and ( 1.80 ± 0.31 ) × 10 - 3 mm2/s respectively and that of 22 left optic nerves was (0.24 ± 0.09) × 10-3,(2.25 ± 0.41) × 10-3,(1.61 ± 0.46) × 10-3,and (1.82 ±0.47) × 10-3mm2/s respectively.The FA of optic nerve in patient group was lower than that of control group (P ＜0.05 ),while the meanλ∥,λ ⊥ and MD values was obviously higher than control group (P ＜ 0.05).There was no significant difference between right and left optic nerves in patient gro up ( P ＞0.05).ConclusionsDTI could detect abnormality and provide information about the pathological process of optic nerve in patients with PCACG.

Objective To establish a rapid,accurate and specific method to detect the common mycobacteria based on multiplex real-time PCR.Methods The dual priming oligonucleotide ( DPO)primers and TaqMan probes labeled with FAM,ROX,HEX or JOE fluoresceins at 5' end and eclipse at 3' end respectively were designed to detect the 16S rRNA of mycobacteria.Both specificity and sensitivity were estimated on multiplex real-time PCR detecting genome DNA from 4 mycobacterial model species.Sixty eight early morning sputum specimens collected from hospitalized patients in the Red Cross Hospital of Hangzhou were detected by multiplex real-time PCR,bacterial culture and smear microscopy simultaneously.The positive rates were analyzed by chi-square.Results Mycobacteria including Mycobacterium tuberculosis and three common non-tuberculosis mycobacteria spp.were identified by multiplex real-time PCR accurately and specifically,with the limited load at 101 cfu/ml.In 68 sputum specimens,31 were positive (positive rate 45.6％ ) by this method,which was significant higher than that by smear microscopy ( positive rate 14.7％,x2 =15.4,P ＜0.05 ).The positive cases were identified as 28 Mycobacterium tuberculosis,1 Mycobacterium avium and 2 Mycobacterium intracellulare in agreement with the culture results.One case,which is detected by culture,but not by PCR,was identified as Mycobacterium chelonae by sequencing.Conclusion The multiplex real-time PCR characterizing as sensitive,specific and time-saving for Mycobacterium tuberculosis and common non-tuberculosis mycobacteria could be chosen as the rapid laboratory test of mycobacterial infection.

Objective:To summarize the main issues, analyze the causes and offer a strategy for AIDS preven-tion and control for all types of healthcare facilities in the health care system. Methods:184 leaders and experts of tertiary-level healthcare facilities ( provinces, cities and counties) from 30 provinces who had been engaged in AIDS prevention and control for more than five years were studied through a questionnaire including the issues and causes in AIDS prevention and control. Results: According to the results of the respondents and experts’ demonstration, the main issues were identified, including a lack of human resources in the system of disease control facilities, a lack of relevant agencies involved in AIDS response, the difficulty for AIDS patients to receive surgery and follow-up, poor detection of HIV, shortcomings in medical insurance, and so on. Conclusion: Based on the analysis of issues and causes, the paper offers strategic solutions: Clearly define the responsibilities of various health institutions in AIDS prevention and control;promote the participation of primary health institutions in AIDS prevention and control; im-prove the medical service system for AIDS patients;improve AIDS medical security policies;reform the management system of antiretroviral therapy drugs;and strengthen technical support from disease control facilities.

Objective To study the relationship between the changes of diffusion tensor imaging(DTI)and diffusion tensor tractography(DTT)of optic radiation and the defect of visual field.Methods DTI and DTT were performed in 37 patients with invasion of the optic radiation by intracranial lesions and in 20 normal control group at 3.0T MR scanner,the visual fields of all cases were examined.The relativity between FA values,ADC values and mean sensitivity(MS)values,mean defect(MD)values of visual field and the relativity between the morphological change of optic radiation and the defect of visual field were analysed.Results In comparison with control group,FA values decreased and ADC values increased in subject group,there were statistically significant differences(P＜0.001)between them.In disordered group,FA values were positive correlation with MS values(r=0.718)and negative correlation with MD values(r=-0.694);ADC values were negative correlation with MS values(r=-0.625)and positive correlation with MD values(r=0.538).DTT showed that the ventral tracts of optic radiation were damaged which oppeared as the defect of ditto quadrant of bilateral field vision in 9 cases,the dorsal tracts and the lateral tracts of optic radiation were damaged which appeared as the defect of inferior quadrant of bilateral field vision in 7 cases,the optic radiations were damaged extensively that caused symmetric or asymmetric congruous hemianopia of bilateral field vision in 21 cases.Conclusion DTI and DTT of optic radiation are intimately correlated with the examined results of visual field,it will be a objective index to extimate the function and the morphological changes of optic radiation.

BACKGROUND:Discectomy and pedicle fixation fusion are golden standard to repair lumbar degenerative disease, but the treatment would induce other complications such as degeneration of adjacent segments or severer pre-existing spinal degeneration. For the problem of lumbar fusion and fixation, lumbar elastic fixation has recently been a hot focus. ＜br> OBJECTIVE:To evaluate the short-term effectiveness of dynamic lumbar pedicle fixation in repair of lumbar spinal stenosis and lumbar disc herniation. ＜br> METHODS:From December 2010 to December 2012, 62 cases of lumbar spinal stenosis and lumbar disc herniation treated with lumbar dynamic system were included. The involved segments included:5 cases at L 3/4 , 20 cases at L 4/5 , 20 cases at L 5 S 1 , 6 cases at double segment L 3/4 and L 4/5, 8 cases at double segment L 4/5 , L 5 S 1 , 3 cases at L 3/4 and L 5 S 1 . There were 34 males and 28 females with an average age of 50.8 years (range 32 to 72 years). According to different fixation systems, they were assigned to three groups:general dynamic lumbar fixation system in 17 cases, K-Rod posterior dynamic stabilization system in 28 cases, and Dynesys system in 17 cases. The fol ow-up time was from 24 to 48 months. Evaluation indexes included visual analogue scale, Oswestry disability index, imaging analysis and excellent and good rate of curative effects. ＜br> RESULTS AND CONCLUSION:Compared with before treatment, visual analogue scale score and Oswestry disability index were significantly improved at 6 months after treatment and final fol ow-up (P＜0.01). No apparent changes were detected in the length of inserted segments and adjacent segments before treatment and during final fol ow-up. There were no significant differences in the excellent and good rate in each group after treatment (P>0.05). These data indicated that the lumbar dynamic system was an effective option for lumbar disc herniation and spinal stenosis. Although there are some differences in the structure of three kinds of flexible fixation, no obvious difference in early therapeutic effects was detected. Long-term effects deserve further investigations.

Objective:To establish the fingerprint analysis method for the aqueous extracts of Eucommia ulmoides from Enshi by HPLC. Methods:The fingerprint of aqueous extracts of ten batches of Eucommia ulmoides from Enshi were analyzed by HPLC. The columnwasWondaSilC18(250mm×4.6mm,5μm). Themobilephaseconsistedofacetonitrile-0.1% phosphoricacidwithgradient elution. The flow rate was 1. 0 ml·min-1 , the detection wavelength was 230 nm, the column temperature was 25℃, and the injection volume was 10 μl. Results:The fingerprint consisted of 12 common peaks. The similarity range of ten batches of Eucommia ulmoides calculated by similarity evaluation system for the chromatographic fingerprint of TCM(Version 2004 A)was 0. 596-0. 997. The standard fingerprint of Eucommia ulmoides was established by HPLC. Conclusion: The established HPLC fingerprint analysis method for Eu-commia ulmoides from Enshi is simple, stable and reproducible, which can effectively control the quality of Eucommia ulmoides from Enshi.