Objective To develop an accurate,rapid,high throughout genotyping method based on oligonucleotide microarray for cytochrome P450 gene polymorphisms related to paclitaxel metabolism.Methods The mutant points of 2C8 * 3,3A4 * 18 and 3A5 * 3C from cytochrome P450 gene were regarded as targets.Based on the sequences in the GenBank,the wild-type and mutant-type probes were specially designed for each mutant point.PCR primers were located in the both sides of mutant point,and furthermore the fragments of PCR products were less than 200 bp.Each type of standard plasmids was constructed.Thus,all the olignucleotide probes were modified with 3'amino-group,and the reverse primers were labeled with fluorescein (Cy3).The probes were immobilized onto certain glass slides.The specific fragments of three genes were amplified and then hybridized with oligonucleotide microarray.The results were analyzed by using certain software.Finally this assay was applied to detect 50 clinical blood specimens.Results When PCR products from standard plasmids were hybridized with DNA microarray,the corresponding probes produced positive signals.Meanwhile,the non-specific hybridization signals did not appear.The results of clinical specimens showed that the mutant rate of CYP2C8 * 3 was 2%.The point of CYP3A4 * 18 for all the clinical specimens was wild-type and the mutant rate of CYP3A5 * 3C was 62%. Meanwhile,the results from detecting 50 clinical blood specimens using oligonucleotide microarray were the same as sequencing analysis.Conclusions Oligonucleotide microarray is a reliable and accurate genotyping assay for cytochrome P450 2C8 * 3.3A4 * 18 and 3A5 * 3C polymorphisms related to paclitaxel simultaneously.This genotyping assay is a high-throughout method for guiding personalized therapy and analyzing metabolism of paclitaxel in vivo.

Objective To investigate the clinical features and bronchoalveolar lavage (BAL)therapy of postinfectious bronchiolitis obliterans (BO) in children. Method Ten children, who had post-infectious BO from February 2009 to February 2010, received BAL therapy, and were retrospectively analyzed. The data included pathology,chnical feature,chest HRCT scan, BALF cellular, levels of blood T cell subtypes and outcome of BAL therapy. Results Adenoviruses or mycoplasma pneumoniae were the most common etiologic agents (4/10, respectively). All patients presented persistent or recurrent dyspneic respirations and wheezing since the initial lung infection. The findings of HRCT included mosaic pattern of perfusion (6/10), accompanied by gas retention,bronchiectasis, atelectasis and bronchial wall thickening. The percentage of neutrophils in BALF was significantly increased in all cases (10/10). There were predominance of CD8+ T cell subtype (9/10) and lower ratio of CD4 +/CD8+ ( 10/10)in blood. Reduced symptoms and shortened hospital stay of BO in 9 of all 10 cases were observed after BAL therapy. Conclusions Severe adenovirus or mycoplasma pneunoniae bronchiolitis and/or pneumonia has higher risk for developing BO in children. Increased percentage of neutrophils in BALF and predominance of CD8 +T cell subtype may play an important role in the mechanism of BO. BAL therepy can reduce the respiratory symptoms of BO in children.

Aim To study the interference role of Bcl-2 siRNA on HL-60 cells. Methods Bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit. The synthesized siRNA was transfected into HL-60 cells with lipid siPORT transfection. Forty-eight hours after the transfection, we used MTT and immunofluorescence to detect cell proliferation and apoptosis,and used RT-PCR and immunofluorescence to detect the level of Bcl-2 mRNA and Bcl-2 protein expression. Result Bcl-2 siRNA reduced the level of Bcl-2 mRNA and Bcl-2 protein expression in HL-60 cells and induced cell apoptosis. There was no difference on the effect of other groups compared with the control. Effective Bcl-2 siRNA specifically degraded Bcl-2 expression in the levels of mRNA and protein and induced HL-60 cells apoptosis.Conclusion These results indicate that siRNA is a highly specific tool for targeted gene knockdown. siRNA-mediated gene silencing is a reliable approach for large-scale screening of gene function and drug target validation.

Objective:To evaluate the application of a medical image software (RadiAnt) in anatomical measurements and precision craniotomy via suboccipital retrosigmoid sinus approach.Methods:A total of 43 inpatients who underwent craniocerebral CT venography (CTV) in our hospital from June 2020 to June 2021 were selected for the study; the CTV data of 35 patients was used to measure the spatial relations between transverse sigmoid sinus junction (TSSJ) and asterion; the preoperative planning in suboccipital retrosigmoid sinus craniotomy with the software was performed in the left 8 patients. Craniotomy time (subjected to exposure of venous sinus margin), venous sinus injury and incidence of complications within 2 weeks of craniotomy in these 8 patients were recorded.Results:(1) Anatomic measurement: for the left side, TSSJ was located at (0.89±0.33) cm lateral and (0.63±0.46) cm inferior to the asterion, and their direct distance was (1.15±0.42) cm; TSSJ was located at (0.76±0.49) cm interior and (1.97±0.52) cm superior to the starting point of the mastoid notch, and their direct distance was (2.18±0.49) cm; about 29% asterion were located superior to the transverse sinus, 37% were located on the surface of the transverse sinus, and 34% were located inferior to the transverse sinus. For the right side, TSSJ was located at (0.88±0.39) cm lateral and (0.64±0.43) cm inferior to the asterion, and their direct distance was (1.12±0.54) cm; TSSJ was located at (0.74±0.40) cm interior and (1.93±0.45) cm superior to the starting point of the mastoid notch, and their direct distance was (2.16±0.43) cm; about 26% asterion were located superior to the transverse sinus, 40% were located on the surface of the transverse sinus, and 34% were located inferior to the transverse sinus. (2) Preoperative planning and surgeries: in these 8 patients, the key-hole was located at (0.96±0.49) cm lateral and (0.53±0.18) cm inferior to the asterion, and (0.46±0.35) cm interior and (1.76±0.47) superior to the starting point of mastoid notch. The interior of sigmoid sinus was located (0.13±0.51) cm interior and (0.21±0.46) cm superior to the starting point of mastoid notch. The inferior of the transverse sinus was located (2.17±0.45) cm interior and (0.53±0.35) cm inferior to the asterion. An accurate localization of the real position of TSSJ, inferior of transverse sinus and interior of sigmoid sinus was performed in all 8 surgical patients. The distance between the margin of the bone window and the interior of sigmoid sinus was (3.5±1.0) mm, and the craniotomy time was (25.7±4.1) min; no sinus injury was noted. Post-operative CT showed good reposition of the bone flaps and less bone defect. There was no cerebrospinal fluid leakage or subcutaneous effusion during the 2 weeks of follow-up.Conclusion:Anatomical measurements and preoperative planning can be quickly finished with low cost with Radiant ?, which provides an efficient and safe method for accurate craniotomy via suboccipital retrosigmoid approach.