1.Construction of shRNA lentivirus vector targeting rat Eap1 gene and Eap1 effects on Kiss1 gene at cellular level
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(4):443-447
Objective·To construct effective shRNA-lentiviral vector targeting rat Eap1 gene and explore the effect of Eap1 on Kiss1 at cellular level. Methods·Four shRNA sequences targeting rat Eap1 mRNA and one negative control sequence were designed and synthesized, and then recombined with lentiviral vectors. After DNA sequencing identification, the shRNA recombinant vectors were co-transfected into 293T cells with pGag/Pol, pRev, pVSV-G to packaging lentiviral particles and the virus titers were determined. NRK-52E cells were transfected with all the four lentiviral-Eap1-shRNAs and the negative control lentivirus, and cells were harvested after 72 h. Real-time PCR and Western blotting was performed to detect the change of the mRNA and protein level of Eap1 gene. The most effective LV-Eap1-shRNA was used to infect NRK-52E cells [non-lentivirus and Eap1-shRNA(-) lentivirus as control groups], and then the level of Kiss1 mRNA expression was detected after infection for 72 h. Results·Seventy-two hours after transfection, the Eap1 mRNA and protein expression between blank group and LV-shRNA(-) group showed no significant difference; while Eap1 expression in all LV-Eapl-shRNA groups reduced significantly in comparing to the control groups. In addition, the lentiviral-Eap1-shRNA4 was the most effective in RNA interference. After blocking Eap1 expression in NRK-52E cells, the Kiss1 mRNA level was significantly increased comparing to the control groups. Conclusion·The effective Eap1-shRNA lentiviral vector is established successfully. The Kiss1 gene expression decreases after blocking Eap1 in NRK-52E cell line.
2.Ethical and Legal Analysis on Genetic Discrimination
Chinese Medical Ethics 2015;(5):675-678
Because the gene right has the attribute of basic human right , as a legal basis for resolving the prob-lem of genetic discrimination , we should confirm its basic legal status .Genetic discrimination has caused the ethi-cal and legal issues in the field of insurance and employment , then put forward the following measures to solve these issues:In the field of insurance , the principle of utmost good faith should be followed to prevent the unscientific use of gene information;In the field of employment , the principle of general prohibition should be combined with the principle of limited exceptions .
3.The application of the theory of cognition, emotion, will and behavior in research integrity educational practice for the medical postgraduate students:based on experiences from United States
Chinese Journal of Medical Science Research Management 2017;30(4):258-262
Objective The study analyzes the inadequacies of the research integrity educational practice for the medical postgraduate students in the present stage of China.Furthermore,the paper puts forward improvement suggestions and clears the direction for further efforts.Methods From the perspective of the educational law about cognition,emotion,will and behavior,the comparative analysis method is used,based on the rich experience in the United States.Results There are inadequacies of the research integrity educational practice for the medical postgraduate students in China,including that students lack knowledge learning motivation,and the pattern of education is simple.There are not enough emotional experience channels and the participation of students.Students lack honesty will training,and the intensity of supervision and punishment is not strong.The honesty recognition mechanism is weak,and cannot be used to guide practice.Conclusions In order to improve the level of the research integrity for the medical postgraduate students in China,we should optimize the system,follow the law of education,strengthen the transmission of knowledge,put the academic honesty education into normal work,enhance the emotional experience,build a participatory mechanism,highlight the positive guide,and increase the intensity of punishment.
4.Advances in the treatment of platinum-resistant ovarian cancer with PARP inhibitors
Journal of International Oncology 2021;48(3):180-183
Poly ADP-ribose polymerase(PARP)inhibitor is a targeted biological agent for the treatment of ovarian cancer, which is mainly suitable for platinum-sensitive ovarian cancer patients. However, data from clinical trials showed that PARP inhibitors have certain therapeutic effect on platinum-resistant or platinum-refractory ovarian cancer patients. It may become a new hope for treatment of ovarian cancer patients in the future.
5.The Induced Differentiation of Hepatic Stem/Progenitor Cells from Human Umbilical Cord Blood
Chenxi WANG ; Li LI ; Yan LIU
Journal of Medical Research 2006;0(12):-
Objective To study the possibility of umbilical cord blood mononuclear cells(UCB MNCs) having polydifferential potention to be induced and differentiated into hepatic stem/progenitor cells.Methods Collecting 32 cases fresh UCB from mature delivery,UCB MNCs were isolated by density-step centrifugal effect and were randomly divided into two groups:(1) control group 8 cases:the basic medium supplemented without cytokines;(2) trial group 24 cases,divided into four groups randomly,and the experimental results were detected at 0d,7d,14d,21d,respectively.The basic medium supplemented with various combinations of growth /differentiation factors(FGF-1,FGF-2,LIF,SCF,HGF,and OSM);cell morphological change was observed,hALBmRNA was examinated using RT-PCR methods,the expressions of ALB、CK18、CK19 were detected by immunohistochemistry.Results In trial group cell number increased,cell volume enlarged,refraction strong,polymorphic cells(mostly oval cells) appeared and obviously attached on the culture dishes.The expression of CK18、CK19、ALB were positive by immunohistochemistry,and the level of hALBmRNA improved along with the culture time.Conclusions UCB MNCs can be induced and differentiated into hepatic stem/progenitor cells in vitro.Human umbilical cord blood mononuclear cells may be a new source of transplantable hepatic stem/progenitor cells and hepatocytes.
6.The status of cancer pain and the usage of complementary and alternative medicine among gynecological cancer pain patients
Ying LI ; Ping LI ; Chenxi LIU
Chinese Journal of Practical Nursing 2015;31(20):1486-1490
Objective To investigate the status of cancer pain and the usage of complementary and alternative medicine among gynecological cancer pain patients.Methods Thirty-one patients with cancer pain were investigated with self-designed questionnaire in the Gynecology Department of the First Hospital of Lanzhou University.Results The average score of age was (51.23 ± 11.34) years old,the youngest patient was 23 years old.During the period of cancer,54.84% (17/31) of the average degree of cancer pain was mild,77.42% (24/31) of the most severe degree of cancer pain was severe,51.61% (16/31) of the patients experienced two or more parts of the pain,75.47% (40/53) of the cancer pain was pelvic pain,72.72% (24/33) was dull pain.The higher the degree of cancer pain was,the worse the subjective sleep quality was.Merely 25.81% (8/31) of the patients used complementary and alternative medicine methods to treatment cancer pain,6 cases of them considered that the methods were effective.Conclusions Patients with gynecological cancer became younger and younger,their cancer pain was severe and mostly was pelvic pain and dull pain.Patients' subjective sleep quality was affected by cancer pain.Complementary and alternative methods might mitigate cancer pain but their usage frequency was low,which was anticipated to be more used in clinic.
7.Analysis of 34 Cases with Laparoscopic Supracervical Hysterectomy
Li YANG ; Chenxi WANG ; Yan LIU
Journal of Medical Research 2006;0(03):-
Objective To evaluate the clinical results of patients submitted to laparoscopic assisted supracervical hysterectomy for benign uterine disease.Methods Women referred for supracervical hysterectomy for benign uterine diseases were divided into laparoscopy group(n=34)or open group(n=30).The operative time,blood loss,complications,morbidity,ileus,wound infections were compared between the two groups.Results In terms of Operative time,both of total operative time and oophosalpingorectomy time,there was no significant difference between laparoscopic group and open group.Less blood loss was found in laparoscopic group than open group.No complications were found in both groups.Conclusions Laparoscopic assisted supracervical hysterectomy is a safer and easier approach to complete supracervical hysterectomy with reduced blood loss and quick convalescence.It can be successfully integrated into a large health maintenance organization/residency-training program.
8.Researchin NANDA-Ⅰ Nursing Diagnoses, NIC Interventions, NOC Outcomes, and NNN Linkages for elderly patients with abdominal external hernia surgery
Chenxi LIU ; Ping LI ; Junfeng KANG ; Ying LI
Chinese Journal of Practical Nursing 2015;31(6):414-417
Objective The purpose of the study was to identify the most frequently used nursing diagnoses,interventions,outcomes and linkages using standardized nursing terminologies for elderly patients with abdominal external hernia surgery.Methods For this study a descriptive design was used.The frequently used NANDA-Ⅰ,NIC,NOC,and NNN linkages were identified through 38 elderly patients with abdominal external hernia surgery who were admitted in hospital from March to August in 2014.Data were analyzed by SPSS17.0.Results The finding showed the most frequently used 5 nursing diagnoses were activity intolerance,impaired tissue integrity,deficient knowledge,chronic pain,and risk for falls,the frequency were 15.51%(38/245),15.51%(38/245),15.10%(37/245),14.69%(36/245) and 14.69%(36/245),the most frequently used 5 nursing outcomes were activity tolerance,ambulation,tissue integrity (skin and mucous membranes),wound healing (primary intention) and knowledge (diet) the frequency were 10.05%(38/378),10.05%(38/378),10.05%(38/378),10.05%(38/378) and 9.79%(38/378); the most frequently used 5 nursing interventions were environmental management (comfort),positioning,exercise therapy (ambulation,) incision site care and environmental management the frequency was all 6.96%(38/546).The most frequently used top 10 NNN linkages were also identified in this study.Conclusions The most frequently used NANDA-Ⅰ,NIC,NOC,and NNN linkages for patients with abdominal external hernia surgery from this study not only offer new knowledge to nurses but also provide scope of evidence-based nursing research in abdominal external hernia clinics.
9.Optimized design of mini-implant diameter and length in IV osteoid through three- dimensional finite element analysis
Guanjun ZHOU ; Chenxi LI ; Xiaoli JIAO ; Yibing LIU ; Lihua SHAN
Chinese Journal of Tissue Engineering Research 2016;20(12):1812-1817
BACKGROUND:Diameter and length of mini-implant have effects on its stability, which has been reported mostly in I and II osteoid, but less in IV osteoid. OBJECTIVE:To optimize the design of mini-implant diameter and length in IV osteoid by a three-dimensional finite element analysis. METHODS:Implant-mandible solid model was established. A 2 N orthodontic force that was perpendicular to the long axis of the implant and at a 30° angle with the distal central axis was applied onto the top of the implant. The implant was designed for different diameters (1.2-2.0 mm) and lengths (6-10 mm). Peak stress and peak displacement of the mandible were mechanicaly assessed, and stress sensitivity variables were analyzed. RESULTS AND CONCLUSION:The stress and displacement of the implant were mainly concentrated in the neck of the implant. The stress of implant-bone interface mainly focused on the contact area of the implant-cortical bone interface, and the stress of the cancelous bone was relatively smal, but the stress of the cortical bone was weakened faster. When the implant length was constant, the implant diameter had a great effect on stress changes, and the stress of bone tissue was reduced with the increase of implant diameter. When the implant diameter was constant, the implant length had no significant effect on the stress of bone tissue. To sum up, the stress of bone tissue and displacement were sensitive to the change of implant diameter rather than the change of implant length. These findings indicate that implant diameter has a greater effect on stress distribution of bone tissue than the implant length, and the implants with > 1.5 mm in diameter are suitable for IV osteoid.
10.Local injection of the microRNA-146a recombinant lentiviral vector into the inner ear for immune-mediated inner ear disease in guinea pigs
Chenxi LUO ; Tao LI ; Changqiang TAN ; He HUANG
Journal of Medical Postgraduates 2016;29(8):801-807
Objective Autoimmune inner ear disease is one of the inner ear diseases that can be effectively treated clinically, and researches on it have significant clinical and practical value.This study explored the immunoregulating and therapeutic effects of the mi-croRNA-146a recombinant lentiviral vector (RLV) on immune-medicated inner ear disease (IMIED) in guinea pigs so as to avoid the adverse reactions of the currently used immunodepressants. Methods IMIED models were established in 30 guinea pigs by immuni-zation with keyhole limpet hemocyanin and divided into three groups: microRNA-146a RLV, empty lentiviral vector ( ELV) control, and surgery simulation control, microinjected via the scala tympani with microRNA-146a RLV, ELV, and PBS, respectively.Before and after immunization and at 7 days after microinjection, the auditory function and the level of specific anti-KLH antibodies in the ser-um were measured by auditory brainstem response ( ABR) audiometry and ELISA, respectively.Seven days after microinjection, 3 ani-mals in each group were subjected to HE staining and light microscopy, another 3 to fluoro-autography for realizing the situations of the inner ear transfected by the lentiviral vector, and the other 4 to measurement of the contents of microRNA-146a in the inner ear tissue. Results Compared with the baseline, immunization significantly increased the level of specific anti-KLH antibodies in the serum (0.09 ±0.01 vs 1.90 ±0.74 in the RLV group, 0.11 ±0.02 vs 2.20 ±0.75 in the ELV group, and 0.11 ±0.02 vs 2.10 ±0.64 in the surgery simulation control) as well as the average threshold of the ABRⅢwave (left ear 11.67 ±2.58 vs 61.67 ±5.16 and right ear 12.50 ±2.73 vs 60.00 ±4.47 in the RLV group;left ear 14.16 ±3.76 vs 64.33 ±9.17 and right ear 13.33 ±2.58 vs 60.83 ± 4.92 in the ELV group;left ear 15.83 ±3.76 vs 64.17 ±10.2 and right ear 15.00 ±5.47 vs 62.50 ±9.35 in the surgery simulation control) .The average threshold of the ABRⅢwave was decreased after local injection into the inner ear as compared with that after immunization, with no statistically significant difference between the ELV and surgery simulation control groups.Fluoro-autographic ob-servation showed that the main parts of the inner ear transfected with microRNA-146a recombinant lentiviruses were the spiral limbus, spiral ganglion afferent fibers, basal cells of the spiral ligament, Corti organ, and psalterial cords.The immune inflammatory reaction of the inner ear was significantly reduced in the RLV group as compared with the ELV and surgery simulation control groups, while the content of microRNA-146a in the inner ear tissue was obviously increased in the former group than in the latter two. Conclusion The microRNA-146a recombinant lentiviral vectors are widely distributed and transfected in the inner ear tissue after injected via the scala tympani.Injecting recombinant microRNA-146a lentiviral vectors into the inner ear can significantly reduce pathological damage and auditory dysfunction caused by inner ear immune inflammatory reaction.