1.Resveratrol induces apoptosis in human primary gastric carcinoma cells
Haibo ZHOU ; Yun YAN ; Jianting CAI ; Qin DU ; Jinming CHEN
Chinese Journal of Pathophysiology 2005;21(7):1340-1344
AIM: To investigate the apoptosis in primary gastric cancer cells induced by resveratrol, and the relation between this apoptosis and expression of bcl- 2 and bax. METHODS: In in vitro experiments, MTT assay was used to determine the cell gowth inhibitory rate. Transmission electron microscopy and TUNEL staining were used to quantitatively and qualitively detect the apoptosis of primary gastric cancer cells before and after the resveratrol treatment. Immunohistochemical staining and RT - PCR was used to detect the expression of apoptosis - regulated gene bcl - 2 and bax. RESULTS: Resveratrol inhibited the growth of primary gastric cancer cells in a dose - and time - dependent manner. Resveratrol induced primary gastric cancer cells to undergo apoptosis with typically apoptotic characteristics. TUNEL assay showed that after the treatment of primary gastric cancer cells with resveratrol for 24, 48, 72, 96 hours, the apoptotic indexs were 4.93% ± 0.19%, 16.74% ± 0.43%, 27.88% ±0.36%, 36.84% ± 1.07 % respectively. Immunohistochemical staining showed that after the treatment of primary gastric cancer cells with resveratrol for 24, 48, 72, 96 hours, the positive rates of Bcl - 2 proteins were 20.68% ± 0.49%, 10.84% ±0.33%, 6.80% ± 0.34%, 3.91% ± 0.15% and the positive rates of Bax proteins were 19.79% ± 0.98%, 30.74% ±0.85%, 40.14% ± 1.17%, 60.08% ± 1.64%. After exposed to resveratrol for 24 h, 48 h, 72 h and 96 h, the density of bcl- 2 mRNA decreased progressively with elongation of time and the density of bax mRNA increased progressively with elongation of time by RT- PCR. CONCLUSION: Resveratrol is able to induce the apoptosis in primary gastric cancer. This apoptosis may be mediated by down- regulation of Bcl- 2 and up- regulation of Bax.
2.Distinct distributions of mouse γδ T ce lls in various tissues and changes after infection
Yuan HU ; Yan LI ; Yun GUAN ; Cai ZHANG
Chinese Journal of Immunology 2016;32(8):1107-1111,1117
Objective:The study focuses on the distinct distributions of γδT cells in various tissues and the changes after Sal-monella typhimurium infection,and attempts to explore the physiological significance of γδT cell distribution and the role of γδT cells in infectious diseases .Methods:Flow cytometry and PCR technique were used to detect the proportion of different γδ T cell subsets among thymus,spleen,lymph nodes,liver,skin,and intestinal intraepithelial lymphocytes .Flow cytometry was applied to detect the secretion of IFN-γand IL-17a.The changes of various γδT cell subsets in liver and intestinal intraepithelial lymphocytes were analyze after Slamonella typhimurium infection.Res ults: γδ T cells were rich in the intestinal epithelium , skin and liver, but poor in the thymus,spleen and lymph nodes .The distribution of different subsets was quite dissimilar .Vγ5+γδT cells chiefly existed in skin ,and Vγ1+,Vγ4+,Vγ7+γδ T cells largely existed in small intestine.γδ T cells in liver mainly secreted IL-17a;however,γδ T cells in intestinal intraepithelial secreted IFN-γ.After infection by Salmonella typhimurium , the proportion of γδ T cells in intestinal intraepithelial increased significantly ,particularly Vγ1 +γδT cells.In Liver,there was no significant change of total γδT cell ratio,but the ratio of Vγ1 +γδT cells reduced ,Vγ4 +γδT cells raised.Conclusi on:γδT cells are rich in the intestinal epithelium ,skin and liv-er.The distribution of different subgroups has specificity .There are large differences in the ability of cytokine secretion among various subgroups of γδT cells.The distribution of γδT cell subgroups in small intestine and liver changes during Salmonella typhimurium in-fection.
3.Analysis on macula injury caused by laser
Yan, ZHANG ; Yan, CAI ; Xiao-Ling, ZHANG ; Yong-Xin, GU ; Xiao-Wei, GAO ; Yun, XIAO
International Eye Science 2016;16(6):1176-1179
?AIM: To discuss the effect of laser exposure on visual acuity and macula.?METHODS: Retrospective and consecutive case series. A retrospective analysis of 11 patients (11 eyes) with laser retinal injury was carried out from January 2014 to June 2015 in Ophthalmology Department of No. 474 Hospital of Chinese PLA. All individuals underwent visual acuity, best corrected visual acuity ( BCVA ) , and spectral-domain optical coherence tomography ( SD-OCT) for macular at first visit, and fundus fluorescein angiography ( FFA ) , visual field, and multifocal electroretinogram ( mf ERG ) were perform if necessary. Symptomatic therapies, supportive therapies and pars plana vitrectomy ( PPV ) were performed depended on the patient’s condition. The patients were followed-up at 1, 3 and 6mo after the first visit, and patients were undertaken visual acuity, BCVA, macular SD-OCT and so on.?RESULTS: Eight patients ( 73%) were under 18 years old and all patients were young males, who were injured by laser pointers when playing. Three patients ( 27%) over 18 years old were injured accidentally at work. Ten (91%) patients’ BCVA were ≤0. 3, while one ( 9%) patient’s BCVA was higher ≥0. 3. Full-thickness macular holes ( the diameter 224-519 μm ) were detected in 10 patients (91%), while sub-foveal RPE changes and IS/OS injury in 1 patient ( 9%) . Macular hole with traction or cystoid edema in 6 eyes (55%) were received PPV, while the other 5 eyes ( 4 eyes with stable macular hole and 1 eyes with RPE injury ) received conservative treatment. Macular hole closed successfully in 1 eye ( 17%) after PPV, while macular hole in the other 5 eyes ( 83%) were stable after PPV of which the cystoid edema faded. The 4 patients with macular hole and 1 patient with RPE injury were stable during follow-up period. However, the BCVA in all patients had no significant improvement at end.?CONCLUSION: Exposure to laser devices could lead to severe macula injury that could reduce central vision, which is permanent.
4.Non-nucleoside reverse transcriptase inhibitors (NNRTIs)resistance-related gene mutations among AIDS patients with antiviral treatment failure in Guangdong province 2015
Yun LAN ; Weiping CAI ; Linghua LI ; Xiaoli CAI ; Yan HE ; Xiaoping TANG ; Fengyu HU ; Junbin LI ; Weilie CHEN ; Liya LI
Chinese Journal of Clinical Infectious Diseases 2017;10(1):14-19
Objective To investigate the prevalence and characteristics of non-nucleoside reverse transcriptase inhibitors (NNRTIs)resistance-related gene mutations among the AIDS patients with virological suppression failure in Guangdong Province 2015.Methods Plasma samples from AIDS patients receiving highly active antiretroviral therapy for more than one year with viral loads > 1000 copies/mL from Guangdong province (except Shenzhen)were collected from January to December 2015.Total 612 HIV-1 gene fragments were amplified from plasma samples using self-developed lab method.Sub-genotypes were determined by phylogenetic tree according to the sequences,NNRTIs resistance-related mutations were determined in Stanford University HIV-1 Drug Resistance Database. The NNRTIs-resistance, the relationships of NNRTIs resistance-related mutations with baseline CD4 +T lymphocyte counts,transmission routes,antiviral regimens and HIV-1 genotypes were analyzed.SPSS 17.0 software was used to analyze the data.Results In 612 patients with virological suppression failure,the main NNRTIs resistance-related mutations were K103 (26.80%),Y181 (14.71 %),V179 (13.73%),G190 (11 .44%) and V106 (10.62%).The susceptibility rate of 310 patients (50.65%)to NNRTIs had changed,the highly resistant rate to nevirapine was 49.51 %,which was higher than that of efavirenz (43.14%),etravirine (5.56%) and rilpivirine (12.25%),respectively,and the differences were statistically significant (χ2 =5.00,296.3 and 198.0,all P <0.05).The incidence rate of drug resistance in patients with baseline CD4 +T lymphocyte counts >200 cells/μL was lower than that in those with baseline CD4 +T lymphocyte counts <200 cells/μL (χ2 =17.93,P <0.01 );the incidence rate of drug resistance was lower in intravenous drug abusers than that of sexually transmitted patients (χ2 =44.21 ,P <0.01 );while the incidence of drug resistance in patients receiving NVP-containing regimens was higher than that in those receiving EFV-containing regimens (χ2 =8.93,P <0.01 ),and the incidence rate was higher in patients with CRF01 _AE than that in those with CRF07_BC and CRF08 _BC (χ2 =8.46 and 8.47,P <0.01 ).Conclusions The results suggest that compliance education and follow-up should be strengthened in patients with high baseline CD4 +T lymphocyte counts and intravenous drug users,and patients with liver diseases should avoid using drugs containing NVP regimens.
5.Survey on the enterovirus 71 survival ability on different surfaces under different climate
Yun CAI ; Lufang JIANG ; Yan SHI ; Yuxin LI ; Qianli WANG ; Liwen JU ; Qingwu JIANG
Chinese Journal of Infectious Diseases 2012;30(7):398-401
Objective To evaluate the survival ability of enterovirus 71 (EV71) on different surface and under different climate.Methods Each 1 × 105 tissue culture infective dose 50 (TCID50)EV71 was added on different aseptic surface of plastic,rubber,cloth and wood,respectively.Then these materials were put into biotron (artificial climatic chamber) which could simulate different temperature and moisture.The viruses were recovered after a definite time and then inoculated into Vero cell.The cytopathic effect (CPE) was observed everyday to survey the survival ability of EV71 on different medium surface.Results The recovery rates of EV71 on medium surface ranged from 89 %-93 %.The survival time of EV71 on medium surface varied under different climatic conditions.The longest survival time of the virus was observed under the condition of 20 ℃ as the temperature and 80% as the humidity.After 24 hours of incubation,the infectious titer on plastic surface reduced about 4 lg.After 72 hours of incubation,the infectious titer reduced at least 3.89 lg on cloth and wood surface.Conclusions Temperature and humidity can affect the survival time of EV71 on medium surface,which is longer in the condition of low temperature and high humidity.The survival ability of EV71 on natural cloth and wood surface is better than that on synthetic plastic surface.
6.Effects of Group Psychological Service on Mental Health of Children Left behind in Rural Areas
Yanling LAN ; Huixiang YOU ; Jingzhi CAI ; Yan LI ; Xiujuan TANG ; Yun ZHOU ; Yiening QIN ; Yanyan ZHANG
Chinese Mental Health Journal 2009;23(11):819-823
Objective:To explore the effect of group psychological service on mental health of the children left behind in rural areas.Methods:Totally 222 children left behind in rural areas were selected randomly and divided into intervention group and control group.The intervention group accepted 8 times of group psychological service.The Eysenck Personality Questionnaire(EPQ),Center for Epidemiological Studies of Depression scale for Children(CES-DC),Rutter Children's Behavior questionnaire for completion by teacher,Piers-Harris Children `s Self-concept Scale(PHCSS)were used in both groups to evaluate the effect at baseline,the end of the 7th intervention,and 6 months after the end of the intervention.Results:After intervention,the intervention group got lower scores than control group in EPQ-N [(8.2±5.8)vs.(10.2±6.1),P<0.05]in the first retest,and in CES-DC [(15.9±7.3)vs.(18.8±7.7),P<0.05]in the second retest.The group gained higher scores in anxietyand happy and satisfaction factors of PHCSS [(6.6±3.4)vs.(5.6±2.7),(6.5±1.9)vs.(5.7±1.8);Ps<0.05]. Conclusion:Group psychological service are effective to children left behind in rural areas with depression emotion and had a role in children's personality remodeling.
7.Effect of suppression of platelet-derived growth factor-α receptor expression with antisense oligonucleotide on proliferation and apoptosis of retinal pigment epithelium cell
Yan-yi, PENG ; Mei-yuan, QIU ; Zhi-xiang, DING ; Miao-yun, LIAO ; Cai-wen, FAN
Chinese Journal of Experimental Ophthalmology 2012;30(4):341-345
BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10% fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.
8.A restrospective analysis of intraoperative cell salvage during cesarean delivery
Haiya YAN ; Yun WU ; Song YE ; Xiaolei CAI ; Chunbo ZHOU ; Guo GAN
Chinese Journal of Anesthesiology 2016;36(11):1297-1301
Intraoperative cell salvage ( IOCS) was conducted during cesarean delivery from May 2011 to April 2016 in our hospital: when the volume of autologous blood collected from the surgical field≥800 ml during surgery and the parturients′hemoglobin≤100 g∕L after hemorrhage, the autologous blood was centrifuged, concentrated, washed and then filtered using a leucocyte depletion filter before reinfusion. IOCS was carried out in 1 085 cases during cesarean delivery within 5 yr. For the parturients who underwent IOCS, the percentage of patients who did not require transfusion of allogeneic red blood cells was 82.95%(900 cases), the percentage of patients who required plasma transfusion was 41.66% (452 cases), the percentage of patients who required platelet transfusion was 3.32% (36 cases), and no autologous blood transfusion?related adverse reactions were found during surgery and hospital stay. In conclusion, IOCS could be safely and effectively used for blood?saving effect in cesarean delivery.
9.The effect of mitochondria-targeted antioxidant peptide SS31 on oxidative damage of lens epithelial cell
Meng, CAI ; Jin, LI ; Jing, LI ; Xiao-yun, CHEN ; Juan, HUANG ; Yan, LUO
Chinese Journal of Experimental Ophthalmology 2013;31(12):1137-1141
Background Presbyopia is one of primary causes affecting the visual and life qualities of the agings,and its mechanism is associated with the oxidative damage of lens epithelial cells with ageing.SS31 is a mitochondria-targeted antioxidant peptide.To study the effect of SS31 on oxidative damage of lens epithelial cells has an important significance for the prevention and treatment of presbyopia.Objective This study was to investigate the effect of SS31 on in vitro oxidative damaged human lens epithelial cells.Methods Human lens epithelial cell line (HLEB-3) was cultured using DMEM with low glucose and 10% fetal bovine serum(FBS).The cell model of oxidative damage was established by adding 200 μmol/L tea-butyl hydropeoxide (t-BHP) into DMEM for 18 hours.The cells were divided into blank control group,t-BHP model group,10 nmol/L SS31 +t-BHP group,100 nmol/L SS31 +t-BHP group,1 μmol/L SS31 +t-BHP group,10 μmol/L SS31 +t-BHP group and 100 pμmol/L t-BHP group,and then MTT assay was used to detect the survival rate of the cells and evaluate the optimal SS31 concentration for sequential study.The cells then were divided into blank control group,t-BHP model group and 1 μmol/L SS31 +t-BHP co-culture group.The change of mitochondrial membrane potential of the cells was tested by JC-1 dye and flow cytometry.Reactive oxygen species (ROS) level in the mitochondria was determined using MitoSOX staining.Results The cell survival rate in the t-BHP model group was (53.42±2.52)%,and that in the blank control group was 100%.The cell survival rate was considerably increased in various concentrations of SS31 groups,showing a significant difference among different groups (F=58.349,P<0.01).A highest survival rate was (82.13 ±3.15) % in the 1 μmol/L SS31 +t-BHP co-culture group,which was statistically significant in comparison with the t-BHP model group (t =28.710,P<0.05).JC-1 dye and flow cytometry assay showed that the ratio between red and green fluorescence intensity was 7.07 ±0.06 in the blank control group,4.46±0.14 in the t-BHP model group and 5.76±0.26 in the 1 μmol/L SS31 +tBHP co-culture group,showing significant difference among the 3 groups (F=172.332,P<0.01).The ratios between red and green fluorescence intensity in the blank control group and 1 μmol/L SS31 +t-BHP co-culture group were higher than that in the t-BHP model (t =2.609,1.303,both at P<0.001).ROS fluorescence cells were much more in the t-BHP model group compared with blank control group and 1 μmol/L SS31 + t-BHP co-culture group.Conclusions SS31 can protect HLEB-3 cells from oxidative stress.SS31 may serve as a potential new approach to the treatment of presbyopia and other age-related diseases of lens.
10.Effects of enteral nutrition with galactooligosaccharides on inflammatory cytokines in rats with severe acute pancreatitis
Yan, ZHONG ; Dong-lian, CAI ; Shan-shan, GENG ; Ting, HAN ; Ling-yun, CHEN
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(6):689-692
Objective To investigate the effects of enteral nutrition with galactooligosaccharides (GOS) on serum inflammatory cytokines in rats with severe acute pancreatitis (SAP). Methods SD rats were randomly divided into sham operation control group, SAP with enteral nutrition (EN) group and SAP with EN supplemented with GOS (GOS-EN) group, and each group was divided into 4 d and 7 d subgroups according to the time that animals were sacrificed (n=8 in each subgroup). Rat SAP models were established by injection of 38 g/L sodium taurocholate beneath the pancreatic capsule. The serum amylase, inflammatory cytokines TNF-α, IL-2 and IL-10 were detected. Results At each time point, the levels of serum amylase in all SAP groups were significantly higher than those in sham operation control group (P < 0.01), and the levels in GOS-EN group were significantly lower than those in EN group (P < 0.01). The levels of serum TNF-α and IL-10 in all SAP groups were significantly higher than those in sham operation control group (P < 0.01), while the levels of IL-2 in all SAP groups were significantly lower than those in sham operation control group. The levels of TNF-α in GOS-EN group were significantly lower than those in EN group (P <0.05), while the levels of IL-2 and IL-10 and the ratio of IL-10/TNF-α were significantly higher than those in GOS-EN group (P < 0.05). Conclusion Early EN supplemented with GOS could modulate the balance of pro- and anti-inflammatory response.