2.The effect of stress environment on the density of bladder pacemaker cells in PDU
Yunfei LI ; Shaofeng ZHANG ; Wei GAN ; Zhenglong ZHANG ; Huihua JI ; Sheng CHEN ; Tianbao WANG ; Yun LIU
The Journal of Practical Medicine 2016;32(19):3151-3154
Objective To investigate the density of bladder pacemaker cells (ICC) in prostatic hyperplasia with detrusor underactivity (PDU) and the impact of stress environment on its growth. Methods 23 PDU guinea pig models and 26 pigs for control (BPH, n = 6; normal group, n = 20) were developed. After water-filled urodynamic pressure measurement of bladder for three groups , detrusor tissue was made into frozen sections. TheICC numberwere measured with immunofluorescence stainingand observed by confocal microscopy. Further ICC cells in vitro were cultured under stress environment and its counts were analyzed by colony formation assay. Results Decreased density of ICC was observed in PDU group (0.28 ± 0.52)% compared to control [(5 ± 0.36)%,P < 0.05] and the BPH group (5.4% ± 0.42)%, P < 0.05). The ratio of dimer and monomer [(6.13 ± 1.45)%] in PDU group was significantly lower than those of the control [31.55 ± 9.67)%, P < 0.05] and the BPH group [(71.52 ± 8.95)%, P < 0.05]. There is a negative correlation between the density of ICC in vitro and duration of stress. By contrast , there is a positive correlation between the density of ICC and incubation time after the stress is removed. Conclusion Distribution of decreased density of ICC is involved in the pathogenesis of PDU , which may be due to long-term stress environment. Thus eliminating stress environment may help restore ICC density in PDU.
4.The measurement of proportion and function of regulatory T cells in unexplained recurrent spontaneous abortion
Yu-Sheng LIU ; Shu-Yun ZHAO ; Jing-Juan JI ; Zhi-Min ZHAI ; Xian- TONG ; Li-Hua LUO ; Hong-Bing LUAN ;
Chinese Journal of Obstetrics and Gynecology 2001;0(03):-
Objective To investigate the proportion and function of CD_4~+ CD_(25)~+ regulatory T cells (CD_4~+ CD_(25)~+ Tr)in unexplained recurrent spontaneous abortion(URSA).Methods(1)Proportion measurement:the proportion of CD_4~+ CD_(25)~+ Tr cells in peripheral blood was measured by double-label flow cytometric analysis.The samples were taken from 15 URSA women,15 normal non-pregnancy women and 13 normal pregnancy women.(2)Function measurement:CD_4~+ CD_(25)~+ Tr ceils and CD_4~+ CD_(25)~+ T ce]ls were extracted from peripheral blood lymphocytes by the microbeads separation.The purity of CD_4~+ CD_(25)~+ Tr cells and CD_4~+ CD_(25)~+ T cells was measured by flow cytometry.The growth inhibitory effect of CD_4~+ CD_(25)~+ Tr cells on CD_4~+ CD_(25)~+ T cells was assessed in vitro.Results The proportion of CD_4~+ CD_(25)~+ Tr cells was decreased significantly in URSA women(6.9?1.8)% than that in normal non-pregnancy women[(10.8?1.1)%] (P0.05).Conclusion The results suggest that decrease in proportion and function of CD_4~+ CD_(25)~+ Tr cells may be associated with URSA.
5.Tramadol and dihydroetorphine produce synergistic analgesic effect and postpones acute opiate tolerance in rats.
Xiao-Yun MING ; Wei WANG ; Ji-Sheng HAN ; Fei LUO
Acta Physiologica Sinica 2005;57(6):696-704
The present study investigated whether a co-application of tramadol (TRA) and dihydroetorphine (DHE) would exert a synergy in analgesic effect and delay acute tolerance development. Intraperitoneal injection of TRA (in mg) and subcutaneous injection of DHE (in ng) were delivered in fixed proportions (1:6.25, 1:12.5, 1:25, 1:50, 1:100, and 1:200). The effect of analgesia was accessed by tail-flick test and analyzed with isobolographic analysis. For test of acute tolerance, six successive injections of either TRA (20 mg/kg) alone, DHE (1 000 ng/kg) alone, or a combination of TRA (20 mg/kg) and DHE (250 ng/kg) were administered. We found that (1) except for 1 mg : 6.25 ng and 1 mg : 50 ng, combinations, all the other ratios produced a significant synergy in their analgesic effect; (2) the effect of analgesia induced by repeated TRA plus DHE injections lasted significantly longer, indicating a slower onset of acute tolerance. These results indicate that TRA and DHE injections in certain dose ratios can induce synergistic analgesia, which is resistant against the development of acute tolerance.
Analgesics, Opioid
;
administration & dosage
;
pharmacology
;
Animals
;
Drug Synergism
;
Drug Tolerance
;
physiology
;
Etorphine
;
administration & dosage
;
analogs & derivatives
;
pharmacology
;
Female
;
Male
;
Pain
;
drug therapy
;
physiopathology
;
Rats
;
Rats, Wistar
;
Tramadol
;
administration & dosage
;
pharmacology
6.Practice of the Teaching Mode Combined Flipped Class with PBL Teaching Method in College English Reading and Writing
Juan WANG ; Yong-Sheng TAO ; Ju-Li SHEN ; Ji-Yun YE ; Yian QU ; Zhen ZHAO
Journal of Kunming Medical University 2018;39(2):128-133
Objective To explore how to apply the teaching mode combined flipped class with PBL teaching method to improve college students' ability of English reading and writing and the effectiveness of classroom. Methods In the class of college English reading and writing, students at grade 2015 were randomly divided into two groups. The experimental group (43 students) was adopted the class teaching with " flipped class and PBL teaching method"and the control group (43 students) was adopted traditional class teaching mode taught by the same teacher. The teaching effects of two groups were studied in a comparative way. The students' final grade, related data in the learning process, students'view of the two teaching modes are studied by analysis of basic data and questionnaire. Results At the end of the semester, the results of the experimental group are better than those of the control group. In the survey of the two class teaching modes, the students in the experimental group spent more time in the learning process than that of the control group.Students think that the teaching mode combined flipped class and PBL teaching method not only improves their autonomic learning ability, classroom participation, the English reading and writing ability, but also helps them perform better in the average scores in the final exam compared to the traditional classroom. Students are more satisfied with the new classroom teaching mode than with traditional one. Conclusion The teaching mode combined flipped class with PBL teaching method can promote the students'autonomous learning, contribute to absorb and internalize knowledge, improve the students'learning interest and learning efficiency, and improve teachers teaching ability.
7.Application of hypobaric ropivacaine spinal anesthesia and nerve block anesthesia in hip surgery ofelderly patients
Chun-Xiang HAO ; Chun-Ji HAN ; Guo-Qing CHEN ; Shu-Yun LIU ; Chang-Sheng ZHANG
Chinese Journal of Tissue Engineering Research 2018;22(7):1009-1013
BACKGROUND: The rational choice of anesthesia for the elderly patients with hip surgery not only ensures the smooth operation, but also significantly reduces the incidence of postoperative complications. OBJECTIVE: To compare the clinical anesthetic effects between combined lumbar plexus-sciatic nerve block and hypobaric ropivacaine spinal anesthesia in hip joint surgery of elderly patients. METHODS: Forty patients who were scheduled for hip joint surgery, at the age of 65-99 years old, American Society of Anesthesiologists grades II-III, were enrolled and randomly allocated to two groups: nerve block (n=20) and spinal anesthesia (n=20). In nerve block group, combined lumber plexus-sciatic nerve block was performed directed by a nerve stimulator under the guidance of ultrasound. In spinal anesthesia group, patients received single-dose hypobaric ropivacaine spinal anesthesia at L3-4interspace. Hemodynamic changes, anesthetic effects and perioperative adverse effects were recorded in both groups before and after anesthesia. RESULTS AND CONCLUSION: (1) The patients' heart rate and mean artery pressure in each group did not change significantly before and after anesthesia in the nerve block and spinal anesthesia groups (P > 0.05). (2) The onset time of anesthesia in spinal anesthesia group was significantly faster than that in nerve block group (P < 0.01). Hypobaric ropivacaine spinal anesthesia had a better analgesic effect during the surgery, which did not need extra intravenous anesthetics. The duration of motor and sense block was significantly longer in nerve block group than in spinal anesthesia group (P < 0.01). However, five patients in spinal anesthesia group needed extra intravenous anesthetics to finish the surgery. (3) No side effects were found in both nerve block and spinal anesthesia groups. (4) These indicated that compared to combined lumbar plexus-sciatic nerve block, hypobaric ropivacaine spinal anesthesia can provide a better analgesic effect during the hip joint surgery with stable hemodynamics. Moreover, hypobaric ropivacaine spinal anesthesia dose not increase the incidence of complications and has very good clinical application prospects.
8.Structural insight into enhanced calcium indicator GCaMP3 and GCaMPJ to promote further improvement.
Yingxiao CHEN ; Xianqiang SONG ; Sheng YE ; Lin MIAO ; Yun ZHU ; Rong-Guang ZHANG ; Guangju JI
Protein & Cell 2013;4(4):299-309
Genetically encoded Ca(2+) indicators (GECI) are important for the measurement of Ca(2+) in vivo. GCaMP2, a widely-used GECI, has recently been iteratively improved. Among the improved variants, GCaMP3 exhibits significantly better fluorescent intensity. In this study, we developed a new GECI called GCaMPJ and determined the crystal structures of GCaMP3 and GCaMPJ. GCaMPJ has a 1.5-fold increase in fluorescence and 1.3-fold increase in calcium affinity over GCaMP3. Upon Ca(2+) binding, GCaMP3 exhibits both monomeric and dimeric forms. The structural superposition of these two forms reveals the role of Arg-376 in improving monomer performance. However, GCaMPJ seldom forms dimers under conditions similar to GCaMP3. St ructural and mutagenesis studies on Tyr-380 confirmed its importance in blocking the cpEGFP β-barrel holes. Our study proposes an efficient tool for mapping Ca(2+) signals in intact organs to facilitate the further improvement of GCaMP sensors.
Calcium
;
chemistry
;
metabolism
;
Calmodulin
;
chemistry
;
genetics
;
metabolism
;
Crystallography, X-Ray
;
Dimerization
;
Green Fluorescent Proteins
;
chemistry
;
genetics
;
metabolism
;
Histidine
;
chemistry
;
genetics
;
metabolism
;
Hydrogen-Ion Concentration
;
Myosin-Light-Chain Kinase
;
chemistry
;
genetics
;
metabolism
;
Peptide Fragments
;
chemistry
;
genetics
;
metabolism
;
Protein Structure, Tertiary
;
Recombinant Fusion Proteins
;
biosynthesis
;
chemistry
;
genetics
9.Analysis of differential gene expression in rat hepatic stellate cells activated by culture or hepatocellular carcinoma cell induction.
Yun-hong XIA ; Zheng-ji SONG ; Rong-xin CHEN ; Sheng-long YE ; Rui-xia SUN ; Qiong XUE
Chinese Journal of Oncology 2009;31(3):164-169
OBJECTIVEHepatic stellate cells (HSC) in hepatocellular carcinoma (HCC) transdifferentiate into extracellular matrix-producing myofibroblasts. Activated HSC can promote invasion and metastasis of HCC. To understand the differences of HSC in normal liver and HCC, we compared the gene expression patterns in HCC cell induction-activated and culture-activated rat HSC.
METHODSHSC were isolated by density centrifugation and exposed to conditioned medium from rat HCC cell line C5F. Expression of 22 012 genes in quiescent HSC, culture-activated HSC and HCC induction-activated HSC was analyzed by cDNA microarray and confirmed by real-time RT-PCR and Western blot.
RESULTS1672 genes were differentially expressed in culture-activated HSC, including proinflammatory factors, cell adhesion molecules, cell surface receptors, signaling transduction molecules and immune factors. 711 genes were differentially expressed in HCC induction-activated HSC. Some of them were identical to those in culture-activated HSC. HCC Induction-activated HSC showed specific gene expression patterns, including Raf1, Rac2, Adam17, Wnt6, MMP-9 and TNF, suggesting that HCC cells can specifically induce HSC activation.
CONCLUSIONThe gene expression patterns in HCC induction-activated HSC are different from those in culture-activated HSC. HCC induction-activated HSC may play a major role in the invasion and metastasis of HCC. In vivo activation should be considered as the standard for the study of HSC biology. HCC induction-activated HSC should be considered as the standard for HSC biology studies.
Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cells, Cultured ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Hepatic Stellate Cells ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Inbred F344
10.Detection of P53 and K-ras gene mutations in lung cancer with oligonucleotide chip.
Jun-Bo LIU ; Ji-Pan XIE ; Zong-Liang ZOU ; Lin-Jie CHEN ; Long-Yun LI ; Sheng-Qi WANG
Chinese Journal of Biotechnology 2002;18(4):447-451
Different factors including hybridization solution components, hybridization temperature, and the concentration and proportion of the labelled primer, which affected the sensitivity and specificity of single mutation identification, were exploited. Asymmetric PCR increased the hybridization sensitivity, and the asymmetric multi-PCR did not affect the specificity, while the sensitivity was improved a little. Among 30 lung cancer samples detected with the oligonucleotide microarray, 12 was found P53 gene mutations and 5 had K-ras gene mutations. The P53 gene mutations identified by the oligonucleotide microarray was proved 80% same as the sequencing results. The obvious statistical relations of K-ras and P53 gene mutations with tumor type, tumor stage and smoking were not obtained because of less samples and mutation sites.
Genes, ras
;
genetics
;
Humans
;
Lung Neoplasms
;
genetics
;
pathology
;
Mutation
;
Oligonucleotide Array Sequence Analysis
;
methods
;
Oligonucleotides
;
genetics
;
Sensitivity and Specificity
;
Tumor Suppressor Protein p53
;
genetics