1.Reversal of adriamycin resistance in human mammary cancer cells by small interfering RNA of MDR1 and MDR3 genes.
Lan, XIAO ; Rui, GAO ; Shi, LU ; Lirong, REN ; Zehua, WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(6):735-7
The purpose of this paper is to investigate the reversal effect of small interfering RNA (siRNA) targeting MDR1 and MDR3 genes on the resistance of MCF-7/ADR cells to adriamycin. siRNA plasmid vector targeting MDR1 and MDR3 genes was transfected into MCF-7/ADR cells, and then was stained with Annexin-V FITC (fluorescein isothiocyanate conjugated) to detect the early stage cell apoptosis by flow cytometry (FCM). 50% inhibition concentration (IC50) of adriamycin for MCF-7/ADR cells was determined by MTT method. MDR1 and MDR3 mRNA was assessed by RT-PCR. Treatment of MCF-7/ADR cells with the two kinds of siRNAs resulted in a reversal of adriamycin resistance of MDR to different extents. 1) The apoptosis efficiency of MDR1 and MDR3 siRNA vector after transfection was (18.21+/-1.65) % and (9.07+/-2.16) % respectively (P<0.05), and there was significant differences in the apoptosis efficiency between pSuppressor Neo vector and the MDR1siRNA or MDR3 siRNA vector (P<0.01); 2) The reversal effect of MDR1 siRNA is higher than that of MDR3 siRNA (P<0.05); 3) The expression of MDRI and MDR3 mRNA can be restrained by pSuppressor Neo MDR1 and MDR3 siRNA respectively, and the reduction in the mRNA level was in a time-dependent manner (P<0.01). MDR1 and MDR3 gene silencing can enhance intracellular adriamycin accumulation in MCF-7/ADR cells, improve sensitivity of MCF-7/ADR cells to adriamycin, and induce cell apoptosis. The reversal effect of adriamycin resistance by siRNA of MDR1 was more effective than that of MDR3.
2.Construction of three-dimensional in vitro culture model of ovarian carcinoma and the study of its multicellular drug resistance.
Meisong, LU ; Rui, GAO ; Lan, XIAO ; Zehua, WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(6):741-3
To explore the role and possible mechanism of apoptosis and caspase-3 activity in the development of multicellular drug resistance of ovary cancer. Ovarian cancer cell A2780 multicellular spheroids (MCS) were obtained from three-dimensional culture. Drug sensitivity of monolayer cells (MC) and MCS were respectively tested by MTT staining and cytometry. The apoptosis of MC and MCS were determined by the flow cytometry (FCM). The expression of bcl-2 and caspase-3 in A2780/MC and A2780/MCS were detected by using Western blot and caspase-3 assay kit. A2780/MC was compacted into mass after 2 days in three-dimensional cell culture model, and MCS had more than two layers of cells growing within 5 days. Compared with A2780/MC, A2780/MCS were more resistant to the anticancer drug, and the apoptosis rate was significantly lower than those of A2780/MC. The activity of caspase-3 in A2780/MCS was significantly lower than the A2780/MC. But the expression of bcl-2 in A2780/MCS was significantly higher than that in A2780/MC. It was suggested that the drug resistance of MCS might be associated with the overexpression of anti-apoptosis protein bcl-2 and the down-regulation of caspase-3 activity.
3.Reversal of multi-drug resistance by vector-based-ShRNA-Mdr1 In Vitro and In Vivo.
Shi, LU ; Qi, HUANG ; Zehua, WANG ; Yinfeng, SONG ; Lijun, WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(5):620-4
In order to investigate the effects of vector-based hairpin small interference RNA (shRNA) on the reversal of multi-drug resistance (mdr) of A2780/Taxol cells, a novel vector pEGFP-H1/mdr1 containing mdr1-shRNA targeting at position 2943-2963 of mdr1 was designed and synthesized. Subsequently, A2780/Taxol cells were transfected with pEGFP-H1/mdr1, and the expression of mdr1 mRNA and P-gp was detected by using RT-PCR and Western blot respectively. MTT was used to measure the 50% inhibition concentration (IC(50)) of Taxol to A2780/Taxol cells. The results showed that at the 24th and 48th h after transfection, the expression of mdr1 mRNA was decreased to (52.1+/-1.0)% and (0.01+/-1.7)%, and that of P-gp decreased to (88.3+/-2.1)% and 0%, respectively. At the 48th h after transfection, the relative reversal rate of A2780/Taxol cells to Taxol was 69.54%. In vivo, the nude mice xenografts were injected with pEGFP-H1/mdr1, and then administrated Taxol. The tumor volume in pEGFP-H1/mdr1-transfected group was significantly reduced as compared with that in blank control group or pEGFP-H1-transfected group (807.20+/-103.16 vs 1563.78+/-210.54 or 1480.78+/-241.24 mm(3), both P<0.01). These results suggested that transfection of pEGFP-H1/mdr1 could efficiently down-regulate the expression of mdr1 mRNA and P-gp in A2780/Taxol cells, and effectively restore the sensitivity of A2780/Taxol cells to Taxol both in vitro and in vivo.
4.Comparison of tracheal intubations using video intubationscope and Macintosh direct laryngoscope in patients with cervical spine immobilization
Zengting LU ; Qianlin YE ; Kangcong ZHANG ; Haoxiang HU ; Zehua TU
China Journal of Endoscopy 2016;22(9):25-29
Objective To compare the clinical efficacy of the video intubationscope and Macintosh direct laryngoscope in simulated cervical spine immobilization. Methods Sixty patients, ASA Ⅰ or Ⅱ , between 19 and 68 years old, underwent general anesthesia requiring oro-tracheal intubation, were randomly assigned to undergo intubation using video intubationscope (group V) or Macintosh direct laryngoscope (group M), 30 cases in each. Each patient was provided mannal in-line axial stabilization of the head and neck by an experienced assistant. The following data were recorded and analyzed: glottic exposure time, Cormark-Lehane grade (C-L classification), tracheal intubation time, total intubation attempts, manoeuvre needed to aid tracheal intubation, failure for tracheal intubation, one-time success rate of tracheal intubation and total success rate of tracheal intubation, mean arterial pressure (MAP) and heart rate (HR) before induction of anesthesia, before intubation, at glottic exposure, at intubation, 1 and 3 min after intubation, and complications. Results Compared with group M, better glottic exposure view (C-L classification) was achieved in group V (P < 0.05), and the tracheal intubation time was shorter (P <0.05), but the glottic exposure time was longer (P < 0.05). More assistance was need and the intubation failure and complication rate was higher in group M (P < 0.05). Compared with T1, MAP in group M were significantly increased at T2~T5 (P < 0.05), MAP in group V were no significantly changed at T2 (P > 0.05) and were significantly increased at T3~T5 (P < 0.05); compared with group M, MAP at T2~T4 in group V were significantly lower (P < 0.05). Compared with T1, HR in group V were no significantly changed at T2~T5, HR in group M were significantly increased at T2~T4 (P < 0.05), and significantly higher than that in group V at the same time point (P < 0.05). Conclusion Compared with Macintosh direct laryngoscopy in patients with cervical spine immobilization, Video intubationscope could provide better view of glottic exposure, decrease the difficulty of intubation and increase the success rate of intubation, have less complications and influence on patient’s hemodynamics.
5.Digital Inventory of PIVAS in Our Hospital
Zehua LUO ; Junhui REN ; Laichun LU ; Desheng MENG
China Pharmacy 2005;0(13):-
OBJECTIVE:To probe into an inventory method of high efficiency and quality for PIVAS.METHODS:Data processing and calculation were carried out using Excel based on the information system of PIVAS.RESULTS & CONCLUSIONS:Digital inventory greatly improves the efficiency and quality of inventory.Under the condition of same workload and same number of staff,working time of digital inventory is 60% less than that of traditional inventory.And consistent rate between material and account is increased by 5%.
6.Comparison of tracheal intubation with video intubationscope and visual laryngoscope in obese patients
Zengting LU ; Kangcong ZHANG ; Zehua TU ; Qianlin YE ; Haoxiang HU
China Journal of Endoscopy 2017;23(5):39-43
Objective To compare the clinical efficacy of orotracheal intubation with video intubationscope and visual laryngoscope in obese patients. Methods 60 ASA I or II obese patients, BMI >30 kg/m2, aged 22 ~ 60 years, underwent elective surgery requiring orotracheal intubation were randomly divided into two groups: the video intubationscope group (Group V) and the visual laryngoscope group (group K), 30 cases in each. Cormark-Lehane grade (C-L classification), tracheal intubation time, total intubation attempts, success rate of tracheal intubation and complications of tracheal intubation were recorded. Results Good glottic exposure view (C-L classification) was achieved in the two groups (P > 0.05), there were no significant difference in tracheal intubation time, the total success rate and the one-time success rate of tracheal intubation between the V and K groups [(24.4 ± 11.6) s vs (22.3 ± 13.2) s, 100.0% vs 100.0%, 90.0% vs 86.7%] (P > 0.05). There was no significant difference in the complications of tracheal intubation between the two groups (P > 0.05). Conclusion Video intubationscope and visual laryngoscope are suitable for tracheal intubation in obese patients, and has an advantage of good glottis exposure view, rapid intubation, great successful rate and few complications.
7.Single operation video intubationscope assisted by mouthpiece in orotracheal intubation
Zengting LU ; Zehua TU ; Haoxiang HU ; Qianlin YE ; Kangcong ZHANG ; Lixun WANG
China Journal of Endoscopy 2016;22(6):9-12
Objective To evaluate the effect of single operation video intubationscope assisted by disposable mouthpiece in orotracheal intubation. Methods 100 patients undergoing general anesthesia were randomly divided into two groups with 50 cases in each: mouthpiece group (group M): single operation video intubationscope assisted by disposable mouthpiece in orotracheal intubation was performed; control group (group C): Video intubationscope oral intubation assisted by helper were applied. The BP, MAP, HR and SpO2 in the two groups were recorded during intubation. The success rate of intubation, duration of glottis exposure, duration of intubation and complications were recorded. Results Oral-tracheal intubation with video intubationscope were successfully completed for all 100 pa-tients, SpO2 during intubation in two groups was maintained above 95.0%, there was no significant hemodynamic changes in two groups. There were no significant difference in the one-time success rate of intubation, duration of glottis exposure and duration of intubation between group M and group C [92.0%vs 88.0%, (13.0 ±7.0) vs (14.0 ±8.0), (20.0 ± 10.0) vs (21.0 ± 11.0), > 0.05]. No significant complications were reported. Conclusion Compared with video intubationscope oral intubation assisted by helper, single operation video intubationscope assisted by dis-posable mouthpiece in orotracheal intubation also is feasible and effective without needing assistant, it is a simple and convenient technology worthy of application.
8.Expression of Survivin, CyclinD1, p21WAF1, Caspase-3 in Cervical Cancer and Its Relation with Prognosis
Shi LU ; Baohua ZHANG ; Zehua WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(1):78-81
The implications of Survivin, CyclinD1, p21WAF1, Caspase-3 in the development, progression and prognosis in cervical cancer were investigated. By using immunohistochemical SP method, the expression of Survivin, CyclinD1, p21WAF1 , Caspase-3 was detected in 41 cases of cervical cancer, 17 cases of cervical intraepithelial neoplasia (CIN) and 10 cases of normal tissues, and their relation with pathological grade, clinical stage, metastasis and survival time was analyzed.The results showed that the positive expression rate of Survivin, CyclinD1 in cervical cancer was significantly higher than in CIN group and normal control group (P<0.05). The median survival time in the patients with cervical cancer positive for Survivin and CyclinD1 was significantly shorter than in those with negative expression (P<0.05). The expression of both Survivin and CyclinD1 was not related with tumor grade, clinical stage and metastasis (P>0. 05). The positive expression rate of p21WAF1 , Caspase-3 in cervical ca rcer was significantly lower than in CIN group and normal control group (P<0.05), and had a close relation with tumor grade (P<0.05). The expression of Survivin in cervical cancer in cervical cancer was negatively associated with that of Caspase-3 (P<0.01), but positively with that of CyclinD1 (P<0.01). Cox Multivariate analysis revealed that Survivin was the independent prognostic indicator influencing the survival time of the patients with cervical cancer (P<0.05). It was suggested that the high expression of Survivin or CyclinD1, and low expression of p21WAF1 or Caspase-3 was closely correlated with the development of cervical cancer. Survivin and CyclinD1 could be used as a useful indicator to predict the prognosis of cervical cancer.
9.A scoping review of application of continuing nursing network platform for discharged premature infants and their caregivers in NICU
Meixian YAO ; Guili WU ; Xiaoxia LU ; Zehua PENG ; Wei CHEN
Chinese Journal of Modern Nursing 2023;29(1):122-127
Objective:To summarize the scope of research on the application of the continuous nursing network platform for premature infants and caregivers discharged from the Neonatal Intensive Care Unit (NICU) , so as to provide reference for future research and practice.Methods:The method of subject words, keywords and Boolean logic was used to retrieve research on the application of NICU's continuous nursing network platform for discharged premature infants and their caregivers in PubMed, Web of Science, Embase, Cochrane Library, China National Knowledge Infrastructure (CNKI) , WanFang Data, VIP and China Biomedical Literature Database. The retrieval time limit was from the establishment of the database to February 14, 2022.Results:A total of 23 articles were included. The content elements of continuous nursing based on network platform included the professional knowledge and nursing skills guidance, peer support, contact with professionals, information and decision-making support, and psychological support. Application forms included mobile applications, website platforms and online videos. The types of outcome indicators were mainly the effect incaregivers and premature infants, the quality of parent-child interaction, breastfeeding and feasibility evaluation. Nursing had positive effect and feasibility.Conclusions:Continuous nursing based on the network platform has achieved positive effects on NICU discharged premature infants and caregivers, and the combination of multiple elements, personalized customization, and multidisciplinary participation are in line with the needs of caregivers. Future research needs to focus on promoting the family function of premature infants, further develop the content of continuous nursing suitable for each child's development stage, explore a safer network service supervision mode and conduct cost-benefit evaluation.
10.The effect of silica in soil on the extraction of biological evidence DNA at the crime scene using the silica bead method
Lu LU ; Zehua GAO ; Tianquan WU ; Liyan YU ; Shenbing GU ; Dongtao JIA
Chinese Journal of Forensic Medicine 2024;39(1):112-114
Objective To study the effect of silica in soil on the extraction of biological evidence DNA at the crime scene using the silica bead method.Methods Mud suspension and diluted blood were mixed to prepare biological samples mixed with dust and soil,which is to simulate biological evidence at the crime scene.Cell lysis was performed using heating lysis and guanidine salt chemical lysis,respectively.DNA was extracted using the silica bead method,amplified by PCR using Identifiler Plus kit and detected by capillary electrophoresis.The electrophoresis results were compared.Using mud suspension instead of silica beads to extract diluted blood DNA to validate the effect of silica in soil on the extraction of biological evidence DNA at crime scene using silica beads method.Results The complete STR loci were obtained after the extraction and amplification of 4 μL,20 μL dilute blood mixed with mud and lysed with heating cracking,whoes average peak heights arel 969.7±376.9 RFU and 9 706.7±349.8 RFU.For the 4 μL dilute blood mixed with mud guanidine salt chemical lysis,it cannot obtain complete STR loci after extraction and amplification.20 μL dilute blood mixed with mud guanidine salt was chemically cleaved and amplified to obtain complete STR loci with an average peak height of 1 899.8±801.3 RFU.After extraction and amplification by mud suspension instead of silica beads to extract 20 μL diluted blood DNA,complete STR loci were obtained.Conclusion Silicon dioxide in soil can bind to DNA in the presence of guanidine salts,leading to a decrease in the efficiency of recovering on-site biological evidence DNA using the silicon bead method.