1.Isolation, culture and in vitro proliferation of breast cancer stem cells after different cycles of neoadjuvant chemotherapy
Wei LIU ; Dongdong WEI ; Lijie HAN
Chinese Journal of Tissue Engineering Research 2015;(36):5806-5810
BACKGROUND:Whether there are breast cancer stem cel microspheres in the breast cancer tissues and whether these microspheres have an impact on isolation and culture of breast cancer stem cel s after different cycles of neoadjuvant chemotherapy are stil unclear. OBJECTIVE:To explore the proliferation and differentiation of breast cancer stem cel s in breast cancer tissues after different cycles of neoadjuvant chemotherapy. METHODS:Breast cancer stem cel microspheres were isolated from the breast cancer tissues after different cycles of neoadjuvant chemotherapy to drawn a cel growth curve. Immunocytochemical method was used to detect ALDH1 expression. RESULTS AND CONCLUSION:Microspheres could be obtained from the specimens of neoadjuvant chemotherapy for two, three and four cycles rather than one cycle. At 3 days prior to culture, there was no difference in the number of cel s isolated after two-and three-cycle neoadjuvant chemotherapy;but after 3 days, the cel s from the three-cycle neoadjuvant chemotherapy proliferated faster than those from the two-cycle neoadjuvant chemotherapy;after 6 days, the cel growth curve of two-cycle neoadjuvant chemotherapy was in the plateau stage, and the proliferation of cel s from the three-cycle neoadjuvant chemotherapy showed a rapid increase trend. The positive expression of ALDH1 in the microspheres from the three-cycle neoadjuvant chemotherapy was higher than that from the two-cycle neoadjuvant chemotherapy. These findings indicate that breast cancer stem cel s from the specimens of two-and three-cycle neoadjuvant chemotherapy both have proliferation and differentiation potentials, and the specimens of three-cycle neoadjuvant chemotherapy or above are preferred.
2.Influence of preoperative biliary drainage on morbidity rate of severely obstructive jaundice patients after pancreaticoduodenectomy
Ning LIU ; Dongdong CHENG ; Longgang WANG
Chinese Journal of Current Advances in General Surgery 2017;20(6):440-443
Objective:To investigate the influence of preoperative biliary drainage (PBD) on morbidity of severely obstructive jaundice patients after pancreaticoduodenectomy (PD).Methods:A total of 98 severely obstructive jaundice(Serum total bilirubin>300 μ mool/L) patients underwent PD between February 2010 and October 2015 were enrolled in the study.The patients were divided into two groups based on undergoing PBD or not.The no-PBD group comprised 52 patients and the PBD group comprised another 46 patients.Perioperatives parameters,including operative time,intraoperative blood loss,postoperative mortality and morbidity and postoperative hospital stay were compared between the two groups.Results:The demographics,preoperative examinations and pathological results were similar between the two groups (P>0.05).Operative time of the no-PBD group was statistically longer than the PBD group (379.44 ± 88.57min vs 346.98 ± 57.17 min,P<0.05).Besides,intraoperative blood loss of the no-PBD group were much more than the PBD group (365.00 ± 187.07mL vs 297.83 ± 139.57 mL,P<0.05).There was no statistical difference of mortality rate between the no-PBD group and the PBD group(3.85% vs 2.17%,P>0.05).The overall morbidity rate of the 2 groups were similar (53.85% vs 43.48%,P>0.05),but the pancreatic fistula rate of no-PBD group was significantly higher than the PBD group (30.77% vs 13.04%,P<0.05).Conclusion:PBD could reduce operative time,intraoperative blood loss and pancreatic fistula rate after PD.Meanwhile,the mortality and overall morbidity rates were similar between the two groups.PBD should be considered for severely obstructive jaundice patients.
3.The functional role of long non-coding RNA PANDAR in promoting colorectal cancer metastasis and its mechanism
Ning LIU ; Dongdong CHENG ; Jinbo JIANG
China Oncology 2017;27(4):268-275
Background and purpose: Accumulating evidence has revealed that long non-coding RNA (lncRNA) is correlated with carcinogenesis and tumor development. Recent literature suggested that lncRNA promoter of CDKN1A antisense DNA damage activated RNA (PANDAR) was involved in the development of various cancers. However, the functional role of PANDAR in colorectal cancer (CRC) has not been elucidated yet. The present study aimed to explore the functional role of lncRNA PANDAR in promoting CRC metastasis and its mechanism.Methods: The expression of lncRNA PANDAR in CRC cell lines and tissues was detected by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR), and the correlation between lncRNA PANDAR expression and CRC clinicopathological characteristics was statistically analyzed. Then, lncRNA PANDAR stably silencing CRC cells (HCT116-shPANDAR), overexpression cells (DLD1-PANDAR) and control vector cells (HCT116-shNC and DLD1-vector) were established using lentiviral vectors. Moreover, Transwell assay and Matrigel assay were performed to investigate the function of lncRNA PANDAR in CRC migration and invasion. Furthermore, the expression of transcriptional factors mediating epithelial-mesenchymal transition of lncRNA PANDAR overexpression cells were monitored by RTFQ-PCR assay, and the function of the target gene in modulating lncRNA PANDAR mediated CRC metastasis was also explored. Results: The expression levels of lncRNA PANDAR in normal colorectal epithelial cells were much lower than in CRC cell. The levels of lncRNA PANDAR in tumor-adjacent tissues were verified to be much lower than in CRC tissues [(171.52±97.80)% vs (100.00±63.18)%, P<0.05]. Moreover, the expression of lncRNA PANDAR was detected to be significantly correlated with CRC TNM stage, lymph node metastasis and distant metastasis (P<0.05). Besides, lncRNA PANDAR deficiency significantly reduced the migration [100.00% vs (42.08±4.77)%, P<0.05] and invasion [100.00% vs (39.14±3.81)%, P<0.05] capabilities in CRC cells, in contrast, the migration [100.00% vs (194.12±9.33)%, P<0.05] and invasion [100.00% vs (204.08±12.27)%, P<0.05] capa-bilities of CRC cells were obviously increased with lncRNA PANDAR overexpression. Furthermore, zinc-finger E-box binding homeobox 1 (ZEB1) expression was detected to be positively correlated with lncRNA PANDAR expression, and ZEB1 silencing could significantly reverse the increased migration and invasion capabilities induced by lncRNA PANDAR in CRC cells. Conclusion: LncRNA PANDAR could promote CRC metastasis by potentially targeting ZEB1. LncRNA PANDAR might be a promising diagnostic marker and therapeutic target for CRC patients.
4.Effects of therapeutic hypercapnia on acute pulmonary allograft rejection induced by macrophages in rats
Can ZHAO ; Dongdong LIU ; Xiaoguang CUI
Chinese Journal of Anesthesiology 2013;33(9):1085-1088
Objective To investigate the effects of therapeutic hypercapnia on acute pulmonary allograft rejection induced by macrophages in rats.Methods Twenty-four adult male Wistar rats and 12 adult male Sprague-Dawley rats,weighing 250-280 g,were used in this study.The recipient rats were randomly divided into 3 groups using a random number table (n =6 each):syngraft group (group S),allograft group (group A) and therapeutic hypercapnia group (group H).In group S,Wistar rats served as donors and recipients,while in A and H groups,Sprague-Dawley rats served as donors and Wistar rats served as recipients.Orthotopic left lung transplantation was performed using the cuff technique.After transplantation,the rats inhaled 50% N2-50% O2 for 90 min during reperfusion in S and A groups,while in group H the rats inhaled N2-O2-CO2 for 90 min during reperfusion and PaCO2 was maintained at 80-100 mm Hg and O2 concentration in inspired air at 48%-50% by adjusting the concentrations of the three gases.At 7 days after operation,the arterial blood sample was collected for blood gas analysis and for determination of serum concentrations of tumor necrosis factor α (TNF-α) and interferon γ (IFN-γ)by ELISA.The oxygenation index was calculated.Then the rats were sacrificed,and the transplanted lungs were removed for microscopic examination and for detection of infiltration of macrophages (by immunohistochemistry)and cell apoptosis (by using TUNEL) in lung tissues.The rejection was scored and apoptotic index was calculated.Results Compared with group S,PaCO2,serum concentrations of TNF-α and IFN-γ,rejection score,the number of macrophages and apoptotic index were significantly increased,and oxygenation index was decreased in group A (P < 0.05).Compared with group A,pH value and oxygenation index were significantly increased,and serum concentrations of TNF-α and IFN-γ,rejection score,the number of macrophages and apoptotic index were decreased in group H (P < 0.05).Conclusion Therapeutic hypercapnia can reduce macrophage-induced acute pulmonary allograft rejection possibly through inhibiting the inflammatory responses and cell apoptosis.
5.The rapid molecular identification of 16 common coagulase negative Staphylococcus
Chaojun LIU ; Dingxia SHEN ; Jing GUO ; Kaifei WANG ; Dongdong LI
Chinese Journal of Microbiology and Immunology 2011;31(12):1129-1132
ObjectiveTo establish the rapid molecular diagnosis of 16 common coagulase negative Staphylococcus(CNS).MethodsDNA sequencing of 16 CNS would be obtained with gap gene.After the alignment gap gene sequences which were available in the GenBank,the bacteria were identified with homological alignment and phylogenetic tree,and compared with the 16S rRNA gene.ResultsThe sequence similarity of the gap sequences ranged from 39% to 98% in 16 CNS.There were the highest similarity (98%) between S.hominis and S.hominis subsp,and the lowest(39% ) between S.saprophyticus and S.xylosus.The sequence similarity of the 16S rRNA sequences ranged from 96 to 98%,at least two species of bacteria similar rate of 99% and the most four species similar rate of 99%.Phylogenetic homology analysis showed that it was a high confidence(99% ) in the detection ofS.xylosus and S.lentus,S.chromogenes and S.intermedius,S.hominis and S.hominis subsp,but for 10 other species of bacteria,gap homology analysis has less unreliable confidence(49%,56% ) and 16S rRNA has more unreliable confidence(43%,43%,50%,56%,63%,65%,76% ).ConclusionAnalysis of gap sequence could identify 16 CNS timely and accurately,with higher confidence than 16S rRNA.
6.Protective effect of rhG-CSF on cognitive function of Alzheimer disease model rats induced by Aβ1-42
Dongdong QIAO ; Hairong WANG ; Lina WANG ; Zengxun LIU ; Jianzhong BI
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(6):494-496
Objective To investigate the effects of rhG-CSF on the improvement of cognitive impairment and anti-apoptosis of Alzheimer disease (AD) induced by Aβ1-42 in rat. Methods Healthy male Wistarirats were randomly assigned to the Aβ group, treatment group and sham operation group. Aβ1-42 (10μg) was injected into bilateral hippocampus to create the rat model of AD. Rats in rhG-CSF group were subcutaneously subjected to 50μg/(kg · d) rhG-CSF for 5 days, while rats in Aβ group were subjected to normal saline. Morris water maze tests were done and expressions of caspase-3 protein were determined by immunohistochemical method on the 7th, 14th, 21st, and 28th day after administration. Results (1) The avoiding latent periods of rhG-CSF group ( ( 34. 33 ±6. 47 ) s, (42. 08 ± 6. 36 ) s, (46. 88 ± 7. 66 ) s, respectively ) were shorter than that of Ap group ((49.79 ±4.87)s, (50.25 ±6.81 )s, (51. 33 ±6.90)s, respectively). The percentages of swimming distances in the target quadrant in rhG-CSF group ( (41.00 ±7.62)% ,(43.33 ±8. 16)% ,(44. 67 ±8.07)% ,respectively) were increased comparing with Ap group((25.33 ±6.89)% , (23. 83 ±4.67)% ,(21.50 ±4.64)% ,respectively). The differences were all statistically significant (P<0.05). (2) Compared with sham operation group,the positive rate of caspase-3 protein in rat's hippocampus of Ap group significantly increased after injecting Aβ1-42 The positive rates of caspase-3 protein in rhG-CSF group on the 7th, 14th day ( (7. 93 ±6. 33) and (8. 83 ±5. 94) were lower than Aβ group ( ( 10.43 ±7. 16) and ( 11. 34 ± 5. 17 ) . The differences were statistically significant (P< 0.05). Conclusion RhG-CSF can improve the cognitive impairment of Alzheimer Disease (AD) induced by Aβ1-42 in rat, decrease the apoptosis of hippocampal neurons and delay the decline of its learning and memory ability to some extent.
7.Study on teaching reformation necessity of medical mycology for undergraduates of medical laboratory specialty
Dingxia SHEN ; Jiyong YANG ; Dongdong LI ; Xiaofeng LIU ; Linjian SONG
Chinese Journal of Medical Education Research 2014;13(11):1115-1118
Objective To study the education of medical mycology for undergraduates of medical laboratory specialty and provide a basis for teaching reformation.Method Setting of mycology related courses of medical mycology for undergraduates in 5 medical schools and 85 inspection and technical personnel's detection of fungi in 81 hospitals were investigated through consultation and questionnaire survey.Results More than 140 class hours for medical mycology were arranged in 5 schools,but as to medical mycology,22 class hours in 1 school and less than 10 class hours in 4 schools,the minimum class hours were 5.Although various numbers of Candida and filamentous fungi could be isolated in hospitals investigated,more than half laboratory workers could not identify penicillium,thermally dimorphic fungi,Zygomycetes and Dematiaceous fungi.Conclusion Education on medical mycology for medical laboratory specialty undergraduates is insufficient and the corresponding teaching lacks such content as medically important pathogenic fungi detection methods and identification characteristics.The hospital technical personnel's fungal identification ability cannot meet the situation of increasing fungal infection involved in clinical medicine,so it is necessary to carry out teaching reformation of medical mycology for undergraduates in laboratory medicine,including adding class hours,increasing course contents and so on.
8.Dynamic Changes of Ephedrine Hydrochloride, Pseudoephedrine Hydrochloride, Amygdalin and Glycyrrhizic Acid in Different Compatibility of San-Ao Decoction
Jinhua ZHANG ; Taoshi LIU ; Jianming CHEN ; Dongdong CHEN ; Haifeng QIAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(11):2312-2317
This paper was aimed to study the dynamic changes of San-Ao(SA) decoction in different compatibility of ephedrine hydrochloride,pseudoephedrine hydrochloride,amygdalin and glycyrrhizic acid.HPLC was used to simultaneously determinate the transfer rate of SA decoction of ephedrine hydrochloride,pseudoephedrine hydrochloride,amygdalin and glycyrrhizic acid.Kromasil C18 (4.6 mm×250 mm,5μm) column was selected with methanol and 0.1% phosphoric acid as a mobile phase to gradient.The flow rate was 1 mL·min-1.The column temperature was 30℃.The injection volume was 10μL.Ephedrine hydrochloride,pseudoephedrine hydrochloride and amygdalin were detected at the wavelength of 208 nm.The glycyrrhizic acid was detected at the wavelength of 250 nm.The results showed that the transfer rate of ephedrine hydrochloride in decoction was more than that of the single preparation.The transfer rate of pseudoephedrine hydrochloride was the highest in the combination of ephedra and bitter almond.The transfer rate of amygdalin was the highest in the combination of ephedra and bitter almond.The transfer rate of glycyrrhizic acid in decoction was more than that of the single preparation.It was concluded that there were dynamic changes in the boiling process during herbal decoction preparation.The effectiveness and stability of Chinese medicine should be improved according to these changes.
9.Relationship and clinical significance between mutated BRAF with prophylactic cen-tral-neck nodal dissection in papillary thyroid carcinoma
Yujie ZHANG ; Baoguo LIU ; Zhiyan ZHAO ; Jindong SHENG ; Dongdong FENG
Journal of Peking University(Health Sciences) 2016;48(3):502-506
Objective:To evaluate the molecular diagnosis marker of papillary thyroid carcinoma (PTC),the relationship between lymphatic metastasis of central neck compartment PTC,and the opera-tion indication of prophylactic central neck dissection.Methods:We conducted a retrospective study, including 275 PTC patients and detected their BRAF mutation rates during 201 2 and 201 4 and explored the risk factors of the central node lymphatic metastasis by Logistic regression model.Results:Of the 275 PTC patients,224 (81 .5%)were female and 51 (1 8.5%)were male.BRAF mutational rates were 53.8% (1 48 /275)and lymphatic metastasis 57.8% (1 59 /275).Multivariate analysis showed calcifica-tion (ORadjusted =1 .47,95%CI:1 .1 0 -1 .98,P =0.01 ),tumor diameter (ORadjusted =1 .48,95%CI:1 .04 -2.30,P =0.048)and age (ORadjusted =1 .48,95%CI:1 .04 -2.30,P =0.048)were associa-ted with lymphatic metastasis.In stratified analysis,BRAF mutation (ORadjusted =3.1 9,95%CI:1 .1 8 -9.43,P =0.023 )in clear boarder group and BRAF mutation (ORadjusted =4.84,95% CI:1 .68 -1 3.84,P =0.003)in calcification group were more likely to have lymphatic metastases.Conclusion:Central neck metastasis takes up a high ratio in papillary thyroid cancer patients,BRAF mutation in pa-pillary thyroid carcinoma is a characteristic molecular event.Furthermore,patients with calcification un-der ultrasound detection,lower age group and longer tumor diameter are more susceptible to suffer central neck metastasis.Especially for stratified analysis,non-calcified BRAF mutation or BRAF mutation with clear border under ultrasound detection are more susceptible to suffer central neck metastasis,and radical prophylactic central neck dissection should be carried on for these patients.
10.Reformation and exploration of selective courses optimization in computer class for medical colleges based on Java language
Xiangkun ZHAO ; Hui CHEN ; Bin JING ; Dongdong LIU
Chinese Journal of Medical Education Research 2015;(8):776-779
Computer selective courses in medical colleges based on C language are facing many problems. First, C language lacks continuity with follow-up courses and well combines with professional courses, which result in loss of interest of students and few students would like choose this selective course. Second, the computer selective courses are miscellaneous and discontinuous, which occupies much time and therefore necessitate integration and optimization. The reformations and implementation schemes are proposed to optimize computer selective courses based on Java language. On the one land, different computer selective courses are optimized, credit hours are compressed, and selective interests of the students are improved. In addition, the practicability and scalability of computer selective courses are enhanced combining the characteristics of medical science specialty.