1.Differentiation of bone marrow mesenchymal stem cells into neurons induced by neural cells by co-culture method
Nailong YANG ; Fen YANG ; Lili XU
Chinese Journal of Tissue Engineering Research 2007;0(29):-
BACKGROUND: There are no studies on differentiation of bone marrow mesenchymal stem cells (BMSCs) into neurons directly induced by neural cells. OBJECTIVE: To establish the co-culture system between BMSCs and neural cells in vitro, and to study the influence of neural cells on the differentiation of BMSCs into neuron in the co-culture system. DESIGN, TIME AND SETTING: The in vitro cytology control experiment was performed at the Central Laboratory of Hospital Affiliated to Medical College of Qingdao University from December 2006 to December 2007. MATERIALS: BMSCS were harvested from bone marrow of patients with diabetes meliuts, who underwent autologous stem cell transplantation at the Hospital Affiliated to Medical College of Qingdao University. Neural cells were collected from brain tissues of infants die of asphyxia during delivery. The third passage of neural cells was used in this study. Transwell double-deck culture dish was purchased from Corning Costar, with a pore diameter of less than 3.0 ?m. Cells could not traverse, but the medium could traverse. METHODS: Neural cells were incubated in Transwell double-deck culture dish at a density of 1?106 in the co-culture group. BMSCs were incubated in the upper layer in LG-DMEM medium for 4-5 days. BMSCs were incubated in both layers in the control group. MAIN OUTCOME MEASURES: The morphological changes of BMSCs were observed and the special markers of neurons cells in BMSCs were examined by immunofluorescence. RESULTS: BMSCs in the co-culture group grew slowly, showing radial processes and connected each other. Positive rate of neuron specific enolase was (32.7?11.5)%. BMSCs in the control group were flat and wide, and negative for neuron specific enolase. CONCLUSION: Microenvironment provided by neural cells promotes the differentiation of BMSCs into neurons.
2.In vitro differentiation and identification of adult bone marrow mesenchymal stem cells into isletsecreting cells
Nailong YANG ; Fen YANG ; Lili XU
Chinese Journal of Tissue Engineering Research 2007;0(38):-
BACKGROUND: There is no ideal method about adult bone marrow mesenchymal stem cells (BMSCs) differentiate into islet-secreting cells and appreciation in vitro at present. Transgene requests strict conditions and complex program. The induction using chemicals is a present-used method. OBJECTIVE: To investigate the culture conditions for inducing adult BMSCs into islet-like cells in vitro. DESIGN, TIME AND SETTING: The cytology in vitro experiment was performed at the Central Laboratory of Affiliated Hospital of Medical College of Qingdao University from January to October 2007. MATERIALS: Bone marrow was collected from diabetic patients undergoing autologous stem cell transplantation at Department of Endocrinology, Affiliated Hospital of Medical College of Qingdao University after signing the informed consent. Epidermal growth factor and basic fibroblast growth factor were obtained from Peprotech Asia. B27 adjunct was purchased from Gibco. Nicotinamide, 2-mercaptoethanol, glutamine and dithizone were bought from Sigma, USA. METHODS: The adult BMSCs were isolated by Percoll from adult bone marrow aspirates and cultured in LG-DMEM, were suspended by Trypsin and passaged for subsequent passages. At the third to fifth passages, BMSCs were incubated at a density of 1?108 L-1 and were induced differentiation into islet-like cells through three developmental stages. In the first stage, BMSCs were incubated in HG-DMEM supplemented with 2-mercaptoethanol, glutamine for 2 days. In the second stage, BMSCs were incubated in HG-DMEM supplemented with epidermal growth factor, basic fibroblast growth factor, B27 and glutamine for 6 days. In the third stage, BMSCs were incubated in HG-DMEM supplemented with nicotinamide and 2-mercaptoethanol for 6 days. BMSCS in the control group were only incubated in the HG-DMEM. MAIN OUTCOME MEASURES: Morphological changes in BMSCs were analyzed under a phase contrast microscopy. Duodenal Homeobox 1 (PDX-1) expression was detected during the second and differentiation stage by immunofluorescence assay. Islet ?-like cell clusters from the 3rd stage were identified by positive dithizone staining. The insulin secretions under the stimulation of high or low glucose were detected by electrochemiluminescence immunoassay. RESULTS: The undifferentiated BMSCs exhibited adherent long spindle-shaped cells. After induction, cells gradually became round and formed clusters. Cells expressed the PDX-1 gene at 8 days and formed islet-like cell clusters that exhibited positive dithizone staining at 14 days. After high and low glucose treatment, no insulin was detected in the control group; insulin content significantly increased at 14 days (t=3.638-9.387, P
3.Cancer-related fatigue in non-small cell lung cancer patients treated with transcutaneous electrical acupoint stimulation during periods of chemotherapy:an experimental pilot study
Yifan WU ; Lili HOU ; Fen GU ; Ying YU
Chinese Journal of Practical Nursing 2017;33(9):693-698
Objective To explore the effectiveness of transcutaneous electrical acupoint stimulation (TEAS) approach in treating patients with cancer- related fatigue during periods of chemotherapy in non-small cell lung cancer (NSCLC) patients in China. Methods A total of 162 participants who treated with GP chemotherapy were randomly assigned to three groups: 66 cases in control group, 61 cases in TEAS group, 67 cases in Sham TEAS group. The following acupoints were used in this study: Qihai (CV 6), Keshu (UB 17), and Zusanli (ST 36). Participants in TEAS group and Sham TEAS group received eight 30-min sessions of TEAS over 28 days. The Revised Piper Fatigue Scale (RPFS) and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 30 (EORTC-QLQ-C30) were used to measure cancer related fatigue (CRF) on the day before chemotherapy days 8 and 28 separately. The differences among three groups were analyzed. Results Finally, 167 patients were included in this study, 56 cases in control group, 57 cases in TEAS group, 49 cases in Sham TEAS group. At the 28th day, the outcomes of the RPFS for TEAS group, Sham TEAS group and control groupscored 2.06 ± 0.90, 2.80 ± 1.34, 3.00 ± 1.29 respectively. There were significantly different among three groups (F=9.784,P<0.01). At the 28th day, the outcomes of the EORTC-QLQ-C30 for TEAS group, Sham TEAS group and control groupscored 64.56 ± 5.00, 54.90 ± 6.25, 54.48 ± 9.68 respectively. There were significantly different among three groups (F=34.119, P<0.01). Conclusions TEAS could help to relived cancer-related fatigue.
4.Study on 746 cases of pituitary adenomas treated with surgery
Minxian SUN ; Muxun ZHANG ; Ting LEI ; Fen ZHOU ; Lili WU ; Sisi YANG ; Chen DONG
Chinese Journal of Endocrinology and Metabolism 2010;26(8):657-661
Objective To analyze the clinical data, including clinical features, treatment, and prognosis,in patients with different kinds of pituitary adenomas. Methods In this retrospective study, 746 cases were included. The characteristics of general epidemiology, clinical symptoms, pathology, imaging, treatment, and prognosis were analyzed. Results Clinical features were different among various pituitary adenomas. Symptoms caused by mass effect and hormone abnormality were expresssed in varying degrees. Serum prolactin>121.28 μg/L can differentiate the prolactin adenoma from the other huge tumor causing hyperprolactinemia due to the mass effect. There is significant relationship between the size and the various types of pituitary adenomas ( P<0.01 ),also between the size and the invasive capability ( P<0.01 ). Conclusions The pituitary adenomas may have their specific epidemiological, clinical, pathological, and imaging features, due to the distinct biological behavior. It is necessary to do the diagnosis, treatment, and prognosis evaluation individually.
5.Codon optimization and eukaryotic expression analysis of the analgesic peptide gene BmK AngM1 from Buthus martensii Karsch.
Jinling YANG ; Lili GAO ; Ping ZHU ; Qi HOU ; Fen WANG ; Wenbo YU ; Tao NIE
Acta Pharmaceutica Sinica 2012;47(10):1389-93
Codon bias is an important factor which influences heterologous gene expression. Optimizing codon sequence could improve expression level of heterologous gene. In order to improve the expression level of BmK AngM1 gene encoding the analgesic peptide from Buthus martensii Karsch in Pichia pastoris, the codon-optimized BmK AngM1 gene according to its cDNA sequence and the preference codon usage of P. pastoris were cloned into expression vector pPIC9K and then transformed into P. pastoris. The expersion of recombinant BmK AngM1 (rBmK AngM1) was inducced by methanol in the medium, and the expression level of the optimized BmK AngM1 gene was 3.7 times of the native one. These results suggested that the expression of BmK AngM1 in P. pastoris could be successfully improved by codon optimization.
6.Nosocomial infections in the surgical intensive care unit—retrospective analysis of five years.
Juan CHEN ; Fen LILI ; Xiangdong GUAN ; Lan LOU ; Minying CHEN ; Changjie CAI ; Zhongzhen LI ; Huiming YI
Chinese Journal of Practical Surgery 2001;21(4):209-212
Objective To evaluate the prevalence of NI in the SICU at our hospital. Methods 181 NI patients in the SICU were retrospectively analysed during Jan 1996~Dec 2000.Results The average NI rate was 9.81%. The major sites of NI were respiratory tract(36.96 %),thoracic/abdominal cavity(25.47 %)and bloodstream infections(9.32 %).The difference in major pathogens of infections in different sites reached statistical significance. For respiratory tract, thoracic/abdominal cavity and bloodstream infections,bacteria were the most common pathogens. Fungi were the moat frequent isolate from urine and stool. Mixed infection proportion was 52.25 %. The most common pathogens were Enterococci, Methicillin resistant Staphylococci、 Pseudomonas Aecruginosa、Escherichia Coli、Candida Albicans and Candida Tropicalis. Conclusions The most common pathogens of NI in SICU are different in different infection sites. The pathogens were complicated and most strains are antibiotics resistant. So it is important to establish NI control and to understand the changes of pathogens so as to prevent the infection.
7.Determination of Polybrominated Diphenyl Ethers and Derivatives, Tetrabromobisphenol A, Hexabromocyclododecane in Egg Samples by Gel Permeation Chromatography and Dispersive Solid Phase Extraction
Minjie LI ; Fen JIN ; Lili YANG ; Rina WU ; Hua SHAO ; Maojun JIN ; Jing WANG
Chinese Journal of Analytical Chemistry 2014;(9):1288-1294
A stimulate method for determination of polybrominated diphenyl ethers ( PBDEs) and derivatives (OH-PBDEs and MeO-PBDEs), tetrabromobisphenol A (TBBPA), hexabromocyclododecane (HBCD) in egg samples was developed by gel permeation chromatography ( GPC) and dispersive solid phase extraction ( DSPE) combined with liquid chromatography tandem mass spectrometric ( HPLC-MS/MS) and gas chroma-tography-negative chemical ionization mass spectrometry ( GC-NCI/MS ) . The analytes were extracted with mixture of hexane and dichloromethane (1∶1, V/V) by accelerated solvent extraction (ASE), and purified by 100 mg C18 dispersive solid phase extraction ( SPE) sorbents followed with gel permeation chromatography (GPC) , and then analyzed by liquid chromatography tandem mass spectrometric (HPLC-MS/MS) and gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS), respectively. The quantita-tion was carried out external standard method. The recoveries of objects were 64. 5%-97. 2% and 65. 6%-109 . 2% ( except BDE85 was 54 . 8%, OH-BDE-137 was 47 . 4%) spiked at 1 . 0 μg/kg or 5 . 0 μg/kg in egg white and egg yolk, respectively. The relative standard deviations (RSDs) were less than 20. 2%. The limits of quantitation (LOQ) for the object were 0. 01-0. 2 μg/kg.
8.The changes of serum interleukin-8 and interleukin-10 in preterm infants delivered from mothers with chorioamnionitis
Zhongxing LU ; Lili LI ; Fen WANG ; Shouling DING ; Yan TENG ; Yueqin GU ; Xiaocheng XU
Chinese Journal of Neonatology 2017;32(3):197-200
Objective To study the changes of serum interleukin-8 (IL-8) and interleukin-10 (IL-10) in preterm infants delivered from mothers with chorioamnionitis (CA) and the possible effects on bronchopuhmonary dysplasia (BPD) and brain injury of preterm infants.Method From October 2014 to October 2015,clinical data from mothers without clinical manifestations of CA giving birth to a preterm baby (gestational age:26 to 33 weeks) were collected in Changzhou Maternal and Child Health Hospital and Suzhou Municipal Hospital.The infants were assigned to CA group and non-CA group according to their mother's placenta pathology.The levels of serum IL-8 and IL-10 were measured using enzyme-linked immunosorbent assay (ELISA) on day 1 and day 7,while the levels of WBC,CRP and PCT were measured at birth.The incidences of BPD and brain injury were also reviewed.Result A total of 67 preterm infants were included,with 51 in the CA group and 16 non-CA group.The levels of IL-8 were significantly higher in the CA group than the non-CA group on both day 1 and day 7 [(21.6 ±9.7) ng/L vs.(7.3 ±2.3) ng/L,(26.6 ± 12.9) ng/L vs.(7.3 ± 2.1) ng/L].The IL-10 levels were significantly lower on day 1 [(1.92±0.17) ng/Lvs.(2.04±0.18) ng/L] and higher on day 7 [(2.42±0.60) ng/L vs.(2.03 ±0.18) ng/L] in the CA group.Significant differences existed on the incidences of BPD (54.9% vs.25.0%) and brain injury (74.5% vs.43.8%) between the two groups (P < 0.05).The levels of CRP and PCT were higher in CA group (P < 0.05) and the WBC was similar between the two groups (P > 0.05).In CA group,on both day 1 and day 7,infants with BPD had similar IL-8 and IL-10 levels comparing with infants without BPD(P > 0.05),also were infants with brain injury comparing with infants without brain injury.Conclusion Chorioamnionitis in pregnant women may affect serum cytokines levels in premature infants and lead to high incidences of BPD and brain injury.
9.Dynamic changes and nursing strategies of chemotherapy-induced nausea and vomiting in patients of newly diagnosed with non-small cell lung cancer
Xiaoyue GAO ; Fen GU ; Lili HOU
Chinese Journal of Modern Nursing 2019;25(1):68-72
Objective? To descriptively analyze the dynamic changes of chemotherapy-induced nausea and vomiting in patients of newly diagnosed with non-small cell lung cancer (NSCLC) during the first chemotherapy cycle after first chemotherapy with gemcitabine plus cisplatin (GP), in order to provide targeted prevention and nursing during the peak period. Methods? A total of 165 NSCLC patients from Shanghai Pulmonary Hospital from January to December 2017 were recruited by convenience sampling method. Chemotherapy-induced nausea and vomiting (CINV) were followed up on the 3rd, 5th, 7th, 14th, and 21st day of chemotherapy, and CINV grades were recorded. Results? In the first chemotherapy cycle, the incidence of nausea was 90.3% (149/165), mostly in grade 2, and the peak period was the 3rd to the 7th day after chemotherapy, while the incidence of vomiting was 50.9% (84/165), mostly in grade 1, and the peak period was the 4th to the 6th days. Conclusions? During the first chemotherapy cycle of GP regimen in patients newly diagnosed with NSCLC, the peak period of CINV was the 3rd to the 7th day, and lasted 3 to 5 days. According to the time window and duration of high incidence of nausea and vomiting, medical staff can intervene in a timely manner to prevent or reduce the occurrence of nausea and vomiting.
10.Expression of Twist in papillary thyroid carcinomas and its roles in differential diagnosis.
Chun-Nian HE ; Lili HE ; Jin Q CHENG ; Shi-Chao CHEN ; Huan-Fen ZHAO ; Jin-Ping ZHAI ; Jing ZHANG
Chinese Journal of Pathology 2008;37(1):35-39
OBJECTIVETo study Twist expression in thyroid papillary carcinoma (PTC) by immunohistochemistry and to assess its usefulness as marker in the differential diagnosis of PTC, follicular adenomas (FA) and benign papillary lesions (BPL).
METHODSFifty cases of PTC, 48 cases of FA and 47 cases of BPL were evaluated using manual tissue chip and SP immunohistochemical stain to detect the expression of Twist and HBME-1, and comparing the staining to that of cytokeratin 19 (CK19).
RESULTSIn PTC, positive rates of Twist, HBME-1 and CK19 were 100% (48/48), 94.0% (47/50) and 78.0% (39/ 50) respectively; in FA, positive rates were 0, 6.7% (3/45) and 0 respectively; in BPL, positive rates were 7.0% (3/34), 2.1% (1/47) and 0, respectively. The differences between PTC and FA and between PTC and BPL were both statistically significant (P = 0. 000). The sensitivity of Twist, HBME-1 and CK19 was 100%, 94.0% and 78.0%; the specifity was 96.4%, 95.7% and 100%; overall accurary was 97.7%, 95.1% and 91.9%, respectively.
CONCLUSIONSPositive rates of Twist is higher than the other markers in PTC. Immunohistochemical staining of Twist has important significance in the differential diagnosis of thyroid lesions. Twist immunohistochemistry maybe helpful in diagnosis and differential diagnosis of PTC.
Adenocarcinoma, Follicular ; metabolism ; Adenocarcinoma, Papillary ; pathology ; Adenoma ; diagnosis ; metabolism ; Biomarkers, Tumor ; immunology ; Carcinoma, Papillary ; diagnosis ; metabolism ; pathology ; Carcinoma, Papillary, Follicular ; metabolism ; Diagnosis, Differential ; Galectin 3 ; genetics ; metabolism ; Immunohistochemistry ; Keratin-19 ; genetics ; Keratins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Thyroid Neoplasms ; diagnosis ; metabolism ; pathology ; Thyroid Nodule ; pathology ; Twist-Related Protein 1 ; genetics ; metabolism