1.Caveolin-1 expression is downregulated by shear stress and TNF-? in human aortic endothelial cells
Qing XIANG ; Xi DONG ; Mei XU ; Ruijuan SUN
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To observe effects of shear stress and TNF-? on caveolin-1 expression. METHODS: Cultured human aortic endothelial cells (HAECs) of passage 3-5 were used in the experiment. Cells were exposed to a laminar flow (shear stress 1.0 Pa) by using a parallel rectangular flow chamber for different time. Caveolin-1 mRNA and protein expression were measured by RT-PCR and Western blot, respectively. Caveolin-1 expression of the cells stimulated by TNF-? were also studied to elucidate the influence of this inflammatory factor. RESULTS: After 24 h of exposure to 1.0 Pa shear stress, both of caveolin-1 protein and mRNA expression decreased in HAECs, especially caveolin-1 mRNA expression (P
2.Inhibitory effect of EGCG on apoptosis of Langerhans cells after UVB irradiation
Xi JI ; Dan LUO ; Xu MIAO ; Xiang-Fei LIN ;
Chinese Journal of Dermatology 1994;0(06):-
Objective To observe the damage to Langerhans cells induced by UVB irradiation,and to evaluate photoprotective effect of these cells from UVB irradiation by epigallocatechin-3-gallate (EGCG).Methods Biopsy specimens were obtained from normal adult foreskin,and epidermal cells were isolated.Density gradient centrifugation and magnetic cell sorting were used simultaneously to purify Langerhans cells from the cell suspension.These cells were then divided into three groups,control (no ir- radiation or EGCG treatment),UVB (irradiation) and EGCG (irradiation+ECCG treatment) groups. The cells in the UVB and EGCG group were irradiated by UVB (30 mJ/cm~2).After the irradiation,the U- VB group was incubated with RPMI-1640 containing 10% bovine serum for 4 hours,while the EGCG group with the same medium containing 200?g/mL of EGCG for 4 hours.Another four hours after the treatment, the cells were collected for the detection of apoptosis rate by propidium iodide staining and flow cytometry. Results Exposure to UVB (30 mJ/cm~2) significantly increased the apoptotic rate of Langerhans cells.The apoptotic rate in EGCG group was significantly lower than that in the UVB group,but was higher than that in the control group.Conclusion Rate of apoptosis of Langerhans cells could be increased by UVB irradia- tion,while EGCG could prevent the increase of apoptosis.
3.The Role and Mechanism of MiR-130b-5p in Down-regulating IGF-1 and Inhibiting the Migration and Invasion of Human Chorionic Trophoblast Cells
Journal of Sun Yat-sen University(Medical Sciences) 2022;43(4):539-547
ObjectiveTo explore the effects of microRNA 130b-5p (miR-130b-5p) on the migration and invasion of human chorionic trophoblast cells (HTR8/SVneo) by targeting insulin-like growth factor-1 (IGF-1). MethodsHTR8/SVneo cells were divided into control group, miR-NC group, miR-130b-5p mimics group, miR-130b-5p mimics+pcDNA3.1 group and miR-130b-5p mimics+pcDNA3.1-IGF-1 group. The dual luciferase reporter gene was used to detect the targeting relationship between miR-130b-5p and IGF-1; real-time fluorescent quantitative PCR was used to detect the expression level of miR-130b-5p and IGF-1in HTR8/SVneo cell samples; the CCK-8 method was used to detect the proliferation of HTR8/SVneo cells; holographic cytometer and Transwell method was used to analyze the migration and invasion abilities of HTR8/SVneo cells; Western blot method was used to detect the expression of IGF-1, invasion, and epithelial-mesenchymal transition (EMT) proteins in HTR8/SVneo cell samples. ResultsThe results of the dual luciferase reporter gene showed that IGF-1 was a potential target gene of miR-130b-5p; compared with the miR-NC group, the expression level of miR-130b-5p, the cell proliferation inhibition rate, and the expression level of E-cadherin increased significantly in the miR-130b-5p mimics group. The expression levels of IGF-1, c-Myc, Cyclin D1, migration distance, number of invaded cells, the expression levels of matrix metalloproteinase 9 (MMP9), matrix metalloproteinase 2 (MMP2), neural cadherin (N-cadherin) and Vimentin were significantly reduced (P<0.05); compared with the miR-130b-5pmimics+pcDNA3.1 group, the cell proliferation inhibition rate and the expression level of E-cadherin in the miR-130b-5p mimics+pcDNA3.1-IGF-1 group were significantly reduced, while the expression levels of c-Myc and CyclinD1, migration distance, number of invasive cells, the expression levels of MMP9, MMP2, Vimentin and N-cadherin were significantly increased (P<0.05). ConclusionThe overexpression of miR-130b-5p may inhibit the migration and invasion of HTR8/SVneo cells by inhibiting the expression of IGF-1.
4.Deepening Teaching Reform of the Fermentation Engineering Experiment to be Adapted to the New Curriculum Reform of Basic Education
Bo XU ; Xiang-Hua TANG ; Zhong-Li WANG ; Yun-Juan YANG ; Zun-Xi HUANG ;
Microbiology 1992;0(05):-
The teaching of the Fermentation Engineering Experiment in normal university must serve for the basic education,placing students' creative spirit and practical ability in the first place.Therefore,teaching reform of the Fermentation Engineering Experiment under the background of new curriculum reform of basic education should be studied from the curriculum content,teaching methodology,training pattern and as-sessment system,in order to cultivate the normal-university students' research ability,working attitude,crea-tive and teaching ability.
5.The use of growth hormone in the treatment of patients with the second degree burns wound
Yong-Xi YANG ; Xiang-Cong XU ; Ze-Guang SUN ; Chu-Zheng YAO ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(08):-
Objective To explore the effects of recombinant human hormone(rhGH) on the plasma total protein,plasma albmin,healing of Wound surfaces in patients with the second degree burns wound.Methods 38 pa- tients with the second degree burns wound were divided into treatment group and control group randomly.All the patients were subject general.19 patients in the treatment group were given rhGH in a dose of 0.2U/kg for 14 days beginning from postoperative 5 days.The plasma total protein concentration,plasma albumin concentration,healing rat of wound surface and scar of patients of the two group were compared.Results The plasma total protein concen- tration plasma albumin concentration of the treatment group were significantly in creased,the scar hyperplasia of the treatment group were significantly mitigated and the healing time of wound surfaces of the treatment group were sig- nificantly shortened.Conclusion rhGH is found to promote protein anabotism and shorten the healing time of wound surfaces and mitigate the scar hyperplasia patients with the second degree burns wound.
6.Effects of serum estradiol levels during controlled ovarian hyperstimulation on outcomes of IVF-ET
Xiang, LU ; Lu, LI ; Xiao-hong, GAO ; Yu, WU ; Bing, XU ; Xiao-xi, SUN
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(7):850-853
Objective To analyse the effects of serum estradiol levels during controlled ovarian hyperstimulation (COH) on the outcomes of in vitro fertilization and embryo transfer (IVF-ET). Methods The clinical data of 472 patients undergoing IVF-ET with GnRH analogues recombinant FSH long protocol were retrospectively analysed. The area under the curve (AUC) of estradiol (E2) level was calculated during COH, and patients were categorized into groups according to the percentile of AUC of E2(AUCE2) during COH. The general characteristics and parameters related to the outcomes of IVF-ET were compared among groups. Results The 10th percentile and 90th percentile of AUCE2 were 3 347.0 pmoL/L and 14 414.3 pmol/L, respectively. Four hundred and seventy-two patients were divided into lower reaction group (AUCE2 3 347.0 pmol/L, n=48), normal reaction group (14 414.3 pmol/L>AUCE2 > 3 347.0 pmol/L, n=376) and higher reaction group (AUCE2≥14 413.3 pmol/L, n=48). There was no significant difference in age, body mass index, baseline follicle stimulating hormone level, time of treatment with gonadotropin, endometrium thickness on day of transfer and embryos transferred(P>0.05). Compared with lower reaction group and normal reaction group, the number of oocytes per retrieval and number of embryos frozen were significantly larger(P<0.01) and the mild/severe ovarian hyperstimulation syndrome rate was significantly higher in higher reaction group(P<0.05). There was no significant difference in fertilization rate, cumulative embryo score, high-grade embryo rate, clinical pregnancy rate and implantation rate among groups (P> 0.05). Conclusion Sustained snpraphysiological serum E2 levels during the COH process do not adversely affect the quality of oocytes and embryos, clinical pregnancy rate and implantation rate to some extent in IVF-ET.
7.Prelimilary experimental study of manganese enhanced-functional MR imaging on cat model about acute epilepsy caused by pentylenetetrazol
Jin-Bai HUANG ; Hai-Bo XU ; Xiang-Quan KONG ; Ding-Xi LIU ; Qing-Xia KONG ; Sheng-Gang SUN ;
Chinese Journal of Radiology 2001;0(01):-
Objective To explore the activated brain region of acute epilepsy in cat model induced by pentylenetetrazol(FFZ)with manganese enhanced-functional MR imaging(ME-fMRI),and evaluate the application of ME-fMRI on localization of the activated brain.Methods Forty cats were divided into 4 groups by random number table method as epileptic A and B groups as well as control A and B groups. Cats of epileptic groups were injected with PTZ(55 mg/kg)intramuscularly,and those of control groups were injected with the saline at same dose.The behavior change in the epileptic and control group A was observed and electroencephalogram(EEG)was also undertaken.Cats of epileptic and control group B were performed ME-fMRI,and the percentage of the enhanced signal intensity was then calculated.Results After injection with PTZ(55 mg/kg)intramuscularly,epileptic seizure was all evoked,and then EEG recording showed spike-wave and polyspike-wave complexes.The neocortex of cats of epileptic group B was diffusely phanero-enhanced on ME-fMRI.The percent enhancement of signal intensity in cortex of frontal lobe,parietal lobe and occipital lobe was(34.6?5.7)% and that in cortex of temporal lobe with(22.9? 6.5)%,whereas those of control group B with(14.9?4.5)% and(11.6?3.2)% respectively.And there was significant difference between the above different localization of the brain in the two groups (t=-10.43,-5.46 respectively,P
8.Potential of vesicular stomatitis virus as an oncolytic therapy for recurrent and drug-resistant ovarian cancer.
Joshua F HEIBER ; Xiang-Xi XU ; Glen N BARBER
Chinese Journal of Cancer 2011;30(12):805-814
In the last decade, we have gained significant understanding of the mechanism by which vesicular stomatitis virus (VSV) specifically kills cancer cells. Dysregulation of translation and defective innate immunity are both thought to contribute to VSV oncolysis. Safety and efficacy are important objectives to consider in evaluating VSV as a therapy for malignant disease. Ongoing efforts may enable VSV virotherapy to be considered in the near future to treat drug-resistant ovarian cancer when other options have been exhausted. In this article, we review the development of VSV as a potential therapeutic approach for recurrent or drug-resistant ovarian cancer.
Animals
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Antineoplastic Agents
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pharmacology
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Apoptosis
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Cell Proliferation
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Drug Resistance, Neoplasm
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Female
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Humans
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Neoplasm Recurrence, Local
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Oncolytic Virotherapy
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methods
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Ovarian Neoplasms
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pathology
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therapy
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virology
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Vesicular stomatitis Indiana virus
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physiology
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Virus Replication
9.Construction of eukaryotic expression vector with brain-derived neurotrophic factor receptor trkB gene.
Tao HUANG ; Xiao-dan JIANG ; Zhong XU ; Jun YUAN ; Lian-shu DING ; Yu-xi ZOU ; Ru-xiang XU
Chinese Journal of Traumatology 2005;8(3):142-146
OBJECTIVETo construct an eukaryotic expression vector carrying rat brain-derived neurotrophic factor receptor trkB gene.
METHODSUsing the total RNA isolated from rat brain as template, the trkB gene was amplified by reverse-transcription-polymerase chain reaction (RT-PCR) with a pair of specific primers which contained the restrictive sites of EcoR I and BamH I. The amplified fragment of trkB gene was digested with EcoR I and BamH I, and then subcloned into cloning vector pMD18-T and expression vector pEGFP-C2 respectively. The recombinant plasmids were identified by restriction endonuclease enzyme analysis and PCR.
RESULTSThe amplified DNA fragment was about 1461 bp in length. Enzyme digestion and PCR analysis showed that the gene of trkB had been successfully cloned into vector pMD18-T and pEGFP-C2.
CONCLUSIONSThe trkB gene of rat has been amplified and cloned into the eukaryotic expression vector pEGFP-C2.
Animals ; Brain-Derived Neurotrophic Factor ; genetics ; pharmacology ; Cloning, Molecular ; methods ; Eukaryotic Cells ; Female ; Gene Expression Regulation ; Genetic Therapy ; methods ; Genetic Vectors ; Male ; Models, Animal ; RNA ; analysis ; Rats ; Rats, Wistar ; Receptor, trkB ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Schwann Cells ; cytology ; Sensitivity and Specificity ; Templates, Genetic ; Transfection
10.Lack of telomerase activity in rabbit bone marrow stromal cells during differentiation along neural pathway.
Zhen-zhou CHEN ; Ru-xiang XU ; Xiao-dan JIANG ; Xiao-hua TENG ; Gui-tao LI ; Yü-xi ZHOU
Chinese Journal of Traumatology 2006;9(4):201-205
OBJECTIVETo investigate telomerase activity in rabbit bone marrow stromal cells (BMSCs) during their committed differentiation in vitro along neural pathway and the effect of glial cell line-derived neurotrophic factor (GDNF) on the expression of telomerase.
METHODSBMSCs were acquired from rabbit marrow and divided into control group, GDNF (10 ng/ml) group. Cytokine.NSCs medium (prepared by our lab, Patent No. ZL02134314. 4) supplemented with 10 percent fetal bovine serum (FBS) was used to induce BMSCs differentiation along neural pathway. Fluorescent immunocytochemistry was employed to identify the expressions of Nestin, neuron-specific endase (NSE), and gial fibrillary acidic protein (GFAP). The growth curves of the cells and the status of cell cycles were analyzed, respectively. During the differentiation, telomerase activities were detected using the telomeric repeat amplification protocol-enzyme-linked immunosorbent assay (TRAP-ELISA).
RESULTSBMSCs were successfully induced to differentiate along neural pathway and expressed specific markers of fetal neural epithelium, mature neuron and glial cells. Telomerase activities were undetectable in BMSCs during differentiation along neural pathway. Similar changes of cell growth curves, cell cycle status and telomerase expression were observed in the two groups.
CONCLUSIONSRabbit BMSCs do not display telomerase activity during differentiation along neural pathway. GDNF shows little impact on proliferation and telomerase activity of BMSCs.
Animals ; Bone Marrow Cells ; enzymology ; Cell Differentiation ; Glial Cell Line-Derived Neurotrophic Factor ; Immunohistochemistry ; Rabbits ; Stromal Cells ; enzymology ; Telomerase ; metabolism