3.Isolation,cultivation and identification of dental pulp stem cells from canine
Journal of Practical Stomatology 2016;32(1):99-103
Objective:To culture canine dental pulp stem cells(cDPSCs)in vitro.Methods:Canine pulp cells were isolated and cultured by enzyme digestion and explanted tissue culture respectively.Cell morphology was observed under phase-contrast micro-scope.The clone forming unit(CFU)of the cells was examined by plate clone formation assay.Cell markers and protein-expression were examined by flow cytometry(FC)and immunofluorescence.Odontogenic and adipogenic potential were evaluated by alizarin red staining and oil red O staining.Results:Short spindle fibroblast-like and steadily growing cells were obtained by both methods.The clone assay showed that CFU was 1 5.1 7% ±2.79%.FC observasion showed that the CD90,STRO-1 and CD24 positive cells were 24.43% ±7.1 0%,20.67% ±1 .42% and 2.03% ±0.06% respectively,but CD34 was negative.Immunofluorescence analysis showed positive expression of Nestin,Vimentin,weak expression of ALP and negative expression of DSP of the cells.Differentiation ex-periment confirmed the odontogenic and adipogenic differentiation potential of the cells.Conclusion:cDPSCs can be cultured in vitro.
5.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.
7.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.
8.Inhibitory Efficiency on Growth in vivo of B16 Melanoma Cell Expressing Angiostatin
Guohong XIA ; Weixin LU ; Li XING ; Jian FEI ; Lihe GUO
Chinese Journal of Cancer Biotherapy 1996;0(04):-
Objective: Study on the growth character of B16 melanoma cell which can express angiostatin. Methods: Angiostatin gene was constructed from human plasminogen cDNA by deletion mutation. A B16 melanoma cell clone named BAG28 which stably expresses angiostatin was established by introducing gene into it. Results: BAG28 in vitro had no changes in proliferation rate and the ability of clone formation in soft agar. Study in vitro showed that the tumor weight had reduced about 87% ( P
9.Research progress of 125I radioactive seed interstitial brachytherapy in the treatment for pelvic malignancies
Tianfan PAN ; Jian LU ; Yong WANG ; Jinhe GUO
Chinese Journal of Radiological Medicine and Protection 2017;37(7):557-561
Brachytherapy with 125I radioactive seed has the advantages of low dose-rate,comtinuous irradiation and repeated implantation to inhibit the replication of tumor cells,and the repeatable implantation.It has been widely used in the treatment for cervial cancer,and can also be applied to pelvic malignancies,including ovarian cancer,colorectal cancer,bladder cancer.Compared with external beam radiotherapy,it can improve the dose of target region to control tumor effectively.At the same time,it can protect the normal tissues and reduce the occurrence of complications.In this paper,we summarized the speciahy,function mechanism,complications and application status of radioactive 125I seed interstitial implantation in the treatment of pelvic malignancies.The application of three dimensional printing in brachytherapy with radioactive 125I seed was also introduced.This review can provide a reliable basis for future standardized seed implantation treatment for pelvic malignancies.
10.Cerebellar cognitive affective syndrome in pediatric patients: clinical analysis of 13 cases.
Xia ZHAO ; Xin-Guo LU ; Jian-Xiang LIAO
Chinese Journal of Contemporary Pediatrics 2007;9(4):387-389
Adolescent
;
Cerebellar Diseases
;
diagnosis
;
drug therapy
;
etiology
;
Cerebellum
;
physiopathology
;
Child
;
Child, Preschool
;
Cognition Disorders
;
diagnosis
;
drug therapy
;
etiology
;
Female
;
Humans
;
Infant
;
Male
;
Mood Disorders
;
diagnosis
;
drug therapy
;
etiology