1.Rapid identification and drug susceptibility assay of mycobacterin in HIV patients
Wenjuan WU ; Guilin DENG ; Jian GUO ; Xueqin QIAN ; Hongzhou LU
Chinese Journal of Microbiology and Immunology 2010;30(2):175-179
Objective To establish the rapid pathogen identification method for HIV and Mycobac-terium tuberculosis (Mtb)co-infection and the assay for the drug susceptibility. Methods Geneprobe and 16S rDNA sequencing were used to differentiate mycobacterium species and modified microscopic observation drug susceptibility (MODS) was used for the drug susceptibility test. The above assays were compared with acid-fast smear, L-J culture and proportional drug susceptibility tests. Results (1) Thirty-four mycobacte-rial isolates were obtained from 112 samples collected from 68 HIV patients. Among these isolates, the strain species were determined by Geneprobe and 16S rDNA sequencing as the followings: 21 Mtb complex, 10 NTM including 5 M.avium complex, 2 M.gordonae, 2 M.kansasii, 1 M.colombiense, and 3 co-infection. (2) The sensitivity of Mtb to rifampicin, ethambutol, isoniazid and streptomycin were 100%, 100%, 76.2%, 90.5% respectively, while the sensitivity of NTM to rifampicin, ethambutol, isoniazid and strepto-mycin were 40%, 60%, 0%, 30% respectively. There is no significant statistic difference between the two methods, MODS and the reference standard, for the drug susceptibility test. (3) Six to eight weeks are nee-ded for the identification of the species of mycobacteria and the drug susceptibility test by using traditional method. In this study, 5-14 d, 6-15 d and 10-14 d are needed for Geneprobe, 16S rDNA sequencing, and MODS respectively. The time for the testing has been dramatically shortened. Conclusion The identifica-tion of mycobacterial species and the drug susceptibility test using clinical samples could be completed within 15 days by using combined Geneprobe, 16S rDNA sequencing and modified MODS. This combined method can be used for the pathogen identification and drug resistant test in HIV patients who are co-infected by my-cobacteria.
2. Study on HPLC characteristic spectra of Lygodii Herba and chemical compositions of different parts from Lygodii Herba
Chinese Traditional and Herbal Drugs 2019;50(11):2708-2811
Objective: To establish the HPLC characteristic spectrum of Lygodii Herba from different habitats in Guangxi Province, and to evaluate the difference of components in different parts of medicinal materials. Methods: HPLC was performed on Waters symmetry C18 (250 mm × 4.6 mm, 5 μm) column, with the mobile phase of acetonitrile-0.2% phosphoric acid at flow rate of 1.0 mL/min; Detection wavelength was 354 nm; The column temperature was 30 ℃ and the sample size was 20 μL. Twelve batches of Lygodii Herba samples were determined and the characteristic spectrum of those were established, and the content of rutin, isoquercetin, and astragalin were determined to evaluate the difference of chemical components in different parts of medicinal materials. Results: There were seven characteristic peaks identified in the characteristic spectra of Lygodii Herba samples. Peak 1 was caffeic acid, peak 2 was Rutin, peak 3 was Isoquercitrin and peak 5 was astragalin. The similarities of 11 batches of samples were proved to be higher than 0.900 and one batch of them was proved to be less than 0.900. The chemical component of stem was richer than that of leaves of Lygodii Herba, and the content of the component was higher in the stem than that of the leaves. Conclusion: The method is simple, accurate, and reproducible, which can provide the scientific evidence for controlling the internal quality standards effectively. The preferred harvest season of Lygodii Herba is autumn and winter.
3.Embolization of dural arteriovenous fitula at the petrous apex region with pressure cooker technique
Guilin LI ; Yongjie MA ; Jian REN ; Hongqi ZHANG
Chinese Journal of Cerebrovascular Diseases 2016;13(7):365-368
Objective To investigate the operation key points and therapeutic effect using the pressure cooker technique (PCT)for embolization of the dural arteriovenous fistula (DAVF)at the petrous apex region. Methods The clinical data of 4 patients with DAVF at the petrous apex region admitted to the Department of Neurosurgery,Xuanwu Hospital,Capital Medical University from October 2015 to February 2016 were analyzed retrospectively. The PCT technique was used to conduct embolization therapy. Results DSA confirmed that 4 patients had DAVF at the petrous apex region with the multi-branch artery blood supply and they were drained via the superior petrosal veins. The patients were embolized by using PCT. DSA confirmed that they were embolized completely after procedure. They were followed up at 3months after surgery and their symptoms were improved. Conclusion For DAVF at the petrous apex region with the multi-branch artery blood supply,the preliminary experience shows that PCT technique may improve the efficiency of embolization,thereby obtaining anatomical cure.
4.Rapid molecular assay and strain typing methods of extrapulmonary tuberculosis
Jian GUO ; Qiwen FAN ; Shuihua LU ; Guilin DENG ; Wenjuan WU
Chinese Journal of Laboratory Medicine 2013;36(7):620-624
[Abstract] Objective To evaluate the effect of Mycobacterium tuberculosis Direct Assay (MTD) for rapid detecting Mycobacterium tuberculosis rRNA and Multi-locus PCR for M.bovis BCG strain typing in patients with suspected extra-pulmonary tuberculosis.Methods From June 2010 to December 2011,47 children and 75 adult patients with suspected extra-pulmonary tuberculosis in Shanghai public health clinical center were recruited.Also 48 non-tuberculosis patients were taken as a negative control.Clinical specimens from these patients were collected.Acid fast stain,solid culture,liquid culture,and MTD were used to detect all clinical specimens simultaneously.Screen tuberculosis strains of the culture isolates by MPT64 antigen assay and use Multi-locus PCR for the BCG strain genotyping of the isolates without MPT64 antigen.SPSS16.0 was used to analyse the results.Results The sensitivity for acid fast stain,solid culture,liquid culture and MTD test was 10.7% (13/122),11.5% (14/122),16.4% (20/122) and 37.7% (46/122),respectively.And the specificity of MTD was 100.0%.Six clinical isolates from children were identified as BCG by Multi-locus PCR typing,the same with chemical tests.Conclusions The MTD assay and the MGIT960 liquid culture are effective and reliable method for diagnosing extra-pulmonary tuberculosis.And Multi-locus PCR can be assisted for the early diagnosis of extra-pulmonary tuberculosis patients with suspected BCG infection.
5.Diagnostic value analysis of 64 layers spiral CT for gastric carcinoma
Zhihua GAO ; Jihua JIAN ; Hongwei REN ; Guilin LIU
Chongqing Medicine 2013;(31):3779-3780,3783
Objective To explore the diagnostic value of 64 layers spiral CT for gastric carcinoma .Methods The 87 patients (primarily diagnosed gastric carcinoma )were studied and scanned under 64 layers spiral CT including plain scans and enhanced scans respectively .The source images were reconstructed using the technique of multilane volume reconstruction (MPVR) and CT virtual endoscopy (CTVE) and conducting TNM stage ,then compared to histopathology results after surgery .Results 85 out of 87 patients with gastric carcinoma were diagnosed by CT .Detection rate of multi-detector CT(MDCT) for gastric cancer was 97 .7% , while the early stage and advanced stage gastric carcinoma were 66 .7% and 100% respectively .The accuracies of 64 layers spiral CT for the T stage ,the N stage and the M stage of the gastric carcinoma were 83 .9% ,72 .4% and 92 .0% respectively .Conclusion 64 layers spiral CT could be helpful for the diagnosis of gastric carcinoma and TNM stage .It might play great roles for the diag-nosis and treatment of gastric carcinoma .
6.Genetic polymorphisms of fifteen short tandem repeat loci in Guilin Han population.
Chinese Journal of Medical Genetics 2009;26(4):465-468
OBJECTIVETo investigate the genetic polymorphisms of 15 short tandem repeat (STRs) in Guilin Han population.
METHODSDNA was extracted by Chelex method, and 15 STRs were analyzed using AmpFISTR Identifiler kit.
RESULTSFour rare alleles, namely FGA * 10, D2S1338 * 10, D3S1358 * 16.2 and D3S1358 * 17.2, were observed. The combined match probability and exclusion probability for the 15 STRs were 2.89 x 10(-17) and 0.9999993, respectively.
CONCLUSIONThese STRs have good discrimination power and exclusion probability in Guilin Han population.
Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Female ; Humans ; Male ; Microsatellite Repeats ; Polymorphism, Genetic
7.Preparation and characterization of linolenic acid-chitosan micelle for doxorubicin oral delivery and its in situ intestinal absorption in rats
Yu-han YANG ; Shi-yuan LIN ; Hui CHEN ; Dian-peng LI ; Jian-fang FENG ; Wei WU ; Wei ZHANG
Acta Pharmaceutica Sinica 2022;57(9):2857-2863
In this study, a novel oral drug delivery system based on linolenic acid-modified chitosan (CS-LA) micelle was developed to improve the oral bioavailability of doxorubicin (DOX), which was proven by its
8.Fe3O4 nanoparticles enhance the sonodynamic therapy effect of chlorin e6 on glioma U251 cells
ZHANG Peng ; CHEN Zihan ; REN Zhongyu ; CHEN Jianjiao ; CHEN Hanren ; WEN Jian
Chinese Journal of Cancer Biotherapy 2021;28(7):702-708
[摘 要] 目的:探讨四氧化三铁(Fe3O4)纳米粒子(PION)作为药物载体增强二氢卟吩e6(chlorin e6,Ce6)在胶质瘤中的增效作用。方法:采用高温降解法和相转移法制备PEG-Fe3O4@Ce6复合纳米粒子(PION@E6),用水合粒径分析、透射电镜、胶体稳定性分析、紫外可见光吸收光谱等方法对PION@E6进行鉴定。CCK-8法检测胶质瘤U251细胞的增殖活性,流式细胞术检测细胞的凋亡水平,DCFH-DA探针法检测细胞中活性氧(reactive oxygen species,ROS)的水平。构建BALB/c-nu裸鼠胶质瘤U251细胞移植瘤模型,动物活体荧光成像术及磁共振成像(MRI)观察PION@E6及Ce6在移植瘤中的潴留时间,比较PION@E6声动力治疗组及Ce6声动力治疗组的第28天生存情况及肿瘤体积。结果:PION@E6的核心粒径为10 nm、水合粒径为(37.86±12.90)nm,具有良好的水溶性和稳定性;吸收光谱及XRD图谱显示Ce6已经负载到Fe3O4纳米粒子上。与Ce6声动力组比较,PION@E6声动力组U251细胞的增殖活性显著下降(P<0.05),细胞凋亡率显著升高(均P<0.05),细胞中ROS水平显著升高(P<0.05)。荷瘤裸鼠胶质瘤U251细胞移植瘤治疗实验结果显示,与Ce6声动力治疗组比较,PION@E6声动力治疗组裸鼠移植瘤组织中潴留时间显著延长(P<0.05),存活的裸鼠数显著增多,移植瘤体积显著缩小(P<0.01)。结论:Fe3O4纳米粒子对Ce6介导的胶质瘤U251细胞声动力治疗具有明显的增效作用。
9.Ultrasmall iron oxide nanoparticles inhibit the migration and invasion of human hepatocellular carcinoma HepG2 cells by enhancing autophagy
CHEN Hanren ; JIANG Shulian ; ZHANG Peng ; REN Zhongyu ; CHEN Jianjiao ; WEN Jian
Chinese Journal of Cancer Biotherapy 2021;28(8):783-789
[摘 要] 目的:探讨超微氧化铁纳米粒子(ultrasmall iron oxide nanoparticle,USIONP)对人肝细胞癌HepG2细胞迁移和侵袭的影响及其可能的机制。方法:采用粒径分析仪和透射电镜分别分析USIONP的水合粒径和核心粒径,Zeta电位和胶体稳定性实验分析USIONP的分散性及其稳定性以鉴定USIONP的成功制备;用不同质量浓度USIONP(0、50、100、200 μg/ml)或200 μg/ml USIONP+5 mmol/L 3-MA(自噬抑制剂)联合处理HepG2细胞,CCK-8法检测HepG2细胞的增殖活力,Transwell法检测细胞的迁移和侵袭能力,WB实验检测自噬标志物Beclin1、LC3、p62的表达,2’,7’-二氯二氢荧光素二醋酸(DCFH-DA)法测定细胞内活性氧(ROS)水平,铁离子比色法检测细胞内铁离子水平。结果:USIONP的平均水合粒径为(37.86±12.90) nm、核心粒径约10 nm,Zeta电位为–23.8 mV,有良好的水溶分散性,证实了USIONP的成功制备。随USIONP质量浓度升高和处理时间延长,HepG2细胞的增殖活力明显降低(均P<0.05);与对照组相比,200 μg/ml USIONP处理HepG2细胞24 h后,迁移、侵袭细胞数量均显著减少(均P<0.05),而3-MA能够部分抵消上述影响(均P<0.05)。与对照组相比,100、200 μg/ml USIONP处理组的HepG2细胞中Beclin1和LC3Ⅱ蛋白相对表达水平均显著升高(均P<0.05),而p62蛋白表达水平下降(均P<0.05);200 μg/ml USIONP可显著提高细胞内ROS水平与铁离子水平,而加入3-MA可阻断其作用(均P<0.05)。结论:USIONP能促进HepG2细胞发生自噬,而自噬通路激活后降解USIONP释放铁离子和导致细胞ROS水平升高,从而抑制HepG2细胞的迁移和侵袭。
10.Feasibility of intentional occlusion of the left subclavian artery in endovascular aortic repair of thoracic aortic disease
Yu ZHANG ; Erping XI ; Jian ZHU ; Shuibo ZHU ; Rongping WANG ; Guihua XU ; Guilin YIN
International Journal of Surgery 2012;39(11):760-762
Objective To investigate the feasibility and effect of intentional occlusion of the left subclavian artery (LSA) in endovascular aortic repair EVAR of thoracic aortic disease.Methods The cases,who needed occlusion of the LSA in EVAR of thoracic aortic disease,were examined cerebral circulation,carotid artery,vertebral basilar artery and circle of Willis.If the vertebral artery had a good blood pathway,and carotid artery and circle of Willis had no stenotic,we occluded the LSA directly in EVAR,and observed the brain and upper limb ischemia complications postoperation.Results Among 40 cases who were occluded the LSA directly in EVAR,28 cases were successful with no neurological complications and left upper limb ischemia symptoms postoperation,12 patients with mild symptoms of subclavian steal syndrome and neurdogical complications,but none required and secondary surgical intervention.Conclusions Prior to intentional LSA occlusion,attention must be paid to potential supraaortic variants and pathologies.Only in this way,it was safe and effective to occlude the LSA in EVAR of thoracic aortic disease.