1.Effect of water soluble chitosan on the growth and phenotype of fibroblasts in vitro
Xiangxin CHE ; Jun CAO ; Weidong LI ; Yidong WANG ; Ping WU
Chinese Journal of Tissue Engineering Research 2007;11(31):6282-6285
BACKGROUND:Studies have demonstrated that chitosan can indirectly promote the proliferation of fibroblast and the synthesis of collagen, Using chitosan and specific fibroblasts together to construct tissue-engineering materials for tendon repair may be an adequate way to promote healing and prevent adhesion during the healing process.OBJECTIVE: To observe the effect of water soluble chitosan (WSC) on the growth and genotype of 3T3TK fibroblasts.DESIGN: Controlled experiment based on observation.SETTING: Jiujiang University Medical College.MATERIALS: 3T3TK fibroblasts were provided by Cell Bank of Chinese Academy of Sciences. WSC (specification: 60 mesh, 30CPS, Jinan Haidebei Marine Bioengineering Co.,Ltd),DMEM (low sugar) (Gibco Company, USA), fetal bovine serum (FBS, Sijiqing Biological Engineering Institute, Hangzhou), penicillin, streptomycin (Biosharp Company, USA),trypsin (BioEev-Tech Scientific & Technical Co.,Ltd, Beijing).METHODS: This experiment was carried out in the Laboratory for Clinical Application of Biology, Center for Medial Scientific Research, Jiujiang University between November 2004 and April 2006. ①Cells in the log phase were digested with 2.5 g/L trypsin to produce single cell suspension and inoculated into a 96-well culture plate at a density of 6 000 cells /200 μL medium. Five groups were prepared, 5 holes per group. 200 μL cell suspension was added into each well.After 24 hours of cultivation, supernatant was discarded, 200 μL DMEM with 1, 0.1, 0.01 and 0.000 1 g/L chitosan was added respectively in the first four groups. The remaining group was taken as the negative control group, in which 200 μL DMEM medium without chitosan was added. The cell suspension was routinely cultured for 72 hours. The cell viability was measured by CellTilter- GloTM Luminescent Cell Viability Assay according to the manufacturer's protocol. Cells in the log phase were split and inoculated into 24-well culture plates. Four groups were prepared (5 holes per group). 1 mL DMEM medium supplemented with 1, 0.1, 0.01 g/L chitosan was added into the first 3 groups respectively, and the 4th group was set as control group. Hydroxyproline and alkaline phosphatase(ALP) kits were used for detecting the contents of collagen and ALP in the supernatant of fibroblasts.MAIN OUTCOME MEASURES : ①Effect of WSC on viability of fibroblasts cultured in vitro.②Contents of collagen and ALP in the cell culture supernatant of fibroblasts.RESULTS:①Detection results of viability of fibroblasts: There were no significant differences in viability of fibroblasts between each chitosan group and control group (P > 0.05).②Contents of collagen and ALP in the cell culture supernatant of fibroblasts: Hydroxyproline content in the cell culture supernatant of fibroblasts of 1 g/L and 0.1 g/L groups was (7.598±0.770) and (8.366±0.308)mg/L, respectively, which was higher than that of control group [(11.269±0.707)mg/L,P < 0.01]; Collagen content in the 1 g/L and 0.1 g/L groups was(56.703±5.748) and(62.437±2.301)mg/L, respectively, which was lower than that of control group [(84.099±5.280)mg/L,P < 0.01]. With the concentration of chitosan increasing, the collagen content was decreased in a dose-dependent manner. There were no significant differences in ALP activity in the cell culture supernatant between each chitosan group and control group (P >0.05).CONCLUSION: WSC does not obviously inhibit the viability of 3T3TK fibroblasts, but can markedly reduce the content of secreted collagen. It is indicated that chitosan can be used as tissue engineering material for tendon repair, and inhibit adhesion formation during tendon repair.
2.Comparison of effects between protease inhibitor and corticosteroid on lung injury
Jun QUE ; Yong MEI ; Xinrong XU ; Quan CAO ; Ping ZHANG
Chinese Journal of Emergency Medicine 2010;19(1):43-46
Objective To study and compare the protective effects of pmtease inhibitor and corticosteroid on endotoxin-indueed acute lung injury in order to guide the choice of appropriate drugs. Method Thirty-two New Zealand rabbits were randomly divided(random number) into four groups with 8 rabbits in each, namely normal controls(C) ; lipopolysaecharide(LPS) group(L) ; ulinastatin(UTI) group(U) and dexamcthasone(DEX) group (D) .Except group C, all rabbits were injected with a dose of LPS 600 μg/kg iv. Meanwhile the rabbits in group U,group D received UTI(100 000 μ/kg), DEX(5 mg/kg), respectively. The specimens were collected 4 hours later for detecting the levels of TNF-α and NO in serum, and blood gas analysis, histological manifestations, the lung wet/dry weight ratio, lung tissue MPO and SOD activity, lung tissue MDA. Data were analyzed by ANOVA (SNK- q test), and P < 0.05 was considered as significantly different. Results Compared with group C, the lungs of the rabbits in group L had inflammatory granulocyte infdtration, diffused alveolar septum thickening and hemorrhagic spots were observed in pathological examinations. The histological changes of group U and group D were much lessened than those in group L. As groups U and D were compared with group L, there were significant differences inmany biomarkers including lung wet/dry weight ratio[(5.02±0.11),(4.93±0.13) vs.(5.37 ±0.29)],lung tissue MPO activity[(0.51 ± 0.05),(0.54±0.07) vs.(0.82 ± 0.09)] and MDA[(0.82 ±0.05),(0.81 ±0.04) vs.(0.96±0.05)], NO[(296.2± 11.7),(291.7 ± 15.8) vs.(351.8±19.6)] and TNF-α[group D(2.021 ± 0.122) vs. group L(4.999 ± 0.139)],lung tissue SOD activity[(120.3 ± 6.1),(122.6±3.5) vs.(105.1 ± 8.5)] and blood gas analysis[pH(7.30±0.23),(7.30±0.17) vs.(7.22±0.45) and PaO_2( 101.9 ± 6.8).( 102.5 ± 4.7) vs.(80.3 ± 3.3)] ; but there were no differences of above mentioned biomarkers between group U and D( P > 0.05). And there were no significant differences in PaCO_2 betweeu group U and D and group L[(37.0 ± 3.3),(37.6 ± 3.0) vs.(34.8 ± 2.3)]( P > 0.05). Conclusions The protective effects of ulinastatin on endotoxin-induced acute lung injury is comparable to those of dexamethasone, thus the former may be a clinical substitute for the latter with less side effects.
3.Effects of alveolar recruitment maneuver on perioperative pulmonary function in morbidly obese patients undergoing laparoscopic sleeve gastrectomy
Ke WEI ; Jun CAO ; Lihua PENG ; Ping LI ; Su MIN
Chinese Journal of Anesthesiology 2016;36(1):26-29
Objective To evaluate the effect of alveolar recruitment maneuver on the perioperative pulmonary function in the morbidly obese patients undergoing laparoscopic sleeve gastrectomy.Methods Forty morbidly obese patients of both sexes,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 18-64 yr,with body mass index ≥ 40 kg/m2,scheduled for elective laparoscopic sleeve gastrectomy,were randomly divided into either control group (group C) or alveolar recruitment maneuver group (group R) using a random number table,with 20 patients in each group.Patients in group C were treated with volume-or pressure-controlled ventilation after creation of pneumoperitoneum,maintaining the peak inspiratory pressure (Ppeak) ≤ 30 cmH2O and partial pressure of end-tidal CO2 35-40 mmHg.Patients in group R received alveolar recruitment maneuver once every 30 min starting from creation of pneumoperitoneum until the end of surgery.Patients were transfered to post-anesthesia care unit (PACU) with endotracheal tube which was extubated when the unified extubation standard was achieved in PACU.The patients who stayed in PACU for 2 h showing no indications for extubation were transfered to intensive care unit for continuous ventilation support.Immediately after intubation,immediately after creation of pneumoperitoneum,at 30,60 and 90 min of pneumoperitoneum,and at the end of pneumoperitoneum,blood samples were collected from the radial artery for blood gas analysis.Immediately after intubation,immediately after creation of pneumoperitoneum,at 30,60 and 90 min of pneumoperitoneum,at the end of surgery,and immediately before discharge from PACU,Ppeak,plateau pressure (Peat),and dynamic lung compliance were recorded.The time for achieving extubation standard and time for achieving the standard for discharge from PACU were recorded.Patients were followed up until discharge,and the feeding time and duration of hospital stay were recorded.Results Compared with group C,PaO2 and oxygenation index were significantly increased at 90 min of pneumoperitoneum,at the end of surgery,and immediately before discharge from PACU,Ppeak was decreased at 60 and 90 min of pneumoperitoneum and immediately after the end of pneumoperitoneum,Pplat was decreased at 60 and 90 min of pneumoperitoneum,the dynamic lung compliance was increased at 30,60 and 90 min of pneumoperitoneum and immediately after the end of pneumoperitoneum,and the time for achieving extubation standard,time for achieving the standard for discharge from PACU,feeding time,and duration of hospital stay were shortened in group R (P<0.05 or 0.01).In group C,one patient did not present with indications for extubation and were transfered to intensive care unit for continuous ventilation support.Conclusion Intraoperative alveolar recruitment maneuver can effectively improve the intraoperative pulmonary function and promote the recovery of postoperative pulmonary function in the morbidly obese patients undergoing laparoscopic sleeve gastrectomy.
4.Community Structure and Ecological Distribution of Airborne Microbes in Autumn in Xi'an
Yi-Ping CHEN ; Ying CUI ; Jun-Ji CAO ;
Journal of Environment and Health 2007;0(10):-
Objective To understand the air microorganisms pollution and the community structure and distribution characteristics of outdoor airborne microbes in Xi'an in autumn.Methods Airborne microbes were sampled by fallen plates method in autumn,2006 in Xi'an at three typical sites,factories region,garden greenbelt region and main traffic region.Results The results showed that the total concentration of airborne microbes at the factories region and garden greenbelt region,main traffic region was 2 500 cfu/m~3,1 354 cfu/m~3 and 4 404 cfu/m~3 respectively.Bacteria and fungal genus were predominant.Gram positive bacteria were much more than Gram negative bacteria.There were 20 genera of bacteria and 8 fungal genus were detected. Micrococcui,Bacillus,Staphylococcus and Pseudornonus were predominant.The most frequently isolated fungal genus were Penicillium,Paecilomces,Cladosporium and Nonsporulatiny mycelia.The community structure of airborne fungal genus and the dominant genus in different function region was significantly different.The distribution characteristic of airborne microbe concentration in different function region was different.The concentration of airborne bacteria was significantly lower in garden greenbelt region than factories region and main traffic region,whereas the concentration of airborne fungi in garden greenbelt region was higher than that in factories region and main traffic region.Conclusion The distribution of the airbome microbes shows a characteristic of time and space,it is affected not only by the environment and pollution,but also by the other factors such as human activities,animals and plants.
5.Changes in expression of CXCR3 in regulatory T cells in renal tissues of mice with renal ischemia-reperfusion injury
Jun CAO ; Ke WEI ; Qingshu LI ; Ping LI ; Jun DONG ; Jie LUO ; Bo CHENG ; Su MIN
Chinese Journal of Anesthesiology 2014;(3):338-340
Objective To evaluate the changes in the expression of CXCR3 in regulatory T cells (Tregs) in the renal tissues of mice with renal ischemia-reperfusion (I/R) injury .Methods Forty-eight SPF male C57BL/6J mice ,aged 8-12 yr ,weighing 20-25 g ,were randomly divided into 3 groups ( n=16 each ) using a random number table:sham operation group (group S) ,group I/R and CD25 monoclonal antibody PC61 group (group P) . Bilateral kidneys were exposed and their pedicles were occluded for 45 min with atraumatic mini-clamp followed by 72 h reperfusion .PC61 250 μg was injected intraperitoneally at 24 h before the model was established .Blood samples were collected from the inferior vena cava at 24 and 72 h of reperfusion (T1 ,2 ) for determination of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations .Bilateral kidneys were obtained for determination of CD4+ CD25+ Foxp3+ Treg count and CXCR3+ CD4+ CD25+ Foxp3+ Treg count in renal tissues and the pathological changes of the kidney were scored .Results Compared with group S , the serum BUN and Cr concentrations and pathological scores were significantly increased at T1 ,2 in I/R and P groups ,and the number of CD4+ CD25+ Foxp3+ Treg and CXCR3+ CD4+ CD25+ Foxp3+ Treg was increased at T2 in I/R group ( P<0.05) .Compared with group I/R ,the serum BUN and Cr concentrations and pathological scores were significantly increased at T2 ,and the number of CD4+ CD25+ Foxp3+ Treg and CXCR3+ CD4+ CD25+ Foxp3+ Treg was decreased at T2 in P group ( P<0.05 ) .Conclusion Up-regulation of CXCR3 is helpful in migration of Tregs into the renal tissues of mice with renal I/R injury .
6.Effect of different doses of propofol on synaptophysin level in hippocampus of mentally depressed rats after electroconvulsive therapy
Yuanyuan LIU ; Su MIN ; Jun DONG ; Jun CAO ; Ping LI ; Yongfeng LIU
Chinese Journal of Anesthesiology 2010;30(1):22-25
Objective To investigate the effects of different doses of propofol on expression of synaptophysin (SYP) mRNA and protein in the hippocampus of mentally depressed rats after electroconvulsive therapy (ECT) .Methods Fifty adult male SD rats weighing 200-250 g were randomly divided into 5 groups (n = 10 each): group Ⅰ control (group C); group D mental depression (group D) ; group Ⅲ , Ⅳ , Ⅴ propofol 90, 110, 130 mg/kg + ECT (group M_1, M_2, M_3). Mental depression was induced by isolation and chronic unpredictable mild stress (CUMS) in group Ⅱ-Ⅴ . Group M_1 , M_2 and M_3 received intraperitoneal (IP) propofol 90, 110 and 130 mg/kg respectively + ECT once a day ×7 consecutive days while group C and D received IP normal saline instead of propofol. The rats underwent open field test and sucrose liquid consumption test the day before and after mental depression was established and one day after treatment. The rats were killed after the last test for determination of expression of SYP mRNA (by RT-PCR) and protein (by immuno-histochemistry) in the hippocampus. Results The ambulation scores, rearing scores, grooming time and sucrose consumption percentage were significantly decreased while the center,grid detention time (CDT) was significantly increased in group Ⅱ-Ⅴ as compared with control group indicating mental depression was successfully induced. The ambulation scores, rearing scores, grooming time, sucrose consumption percentage, SYP mRNA and protein expression were significantly increased while CDT was significantly decreased after treatment in group M_1 and M_2 as compared with group D indicating that propofol 90 or 110 mg/kg combined with ECT was effective for the treatment of mental depression.Conclusion Propofol 90 or 110 mg/kg combined with ECT is effective in treating mental depression while proprofol 130 mg/kg combined with ECT is not. Excessive inhibition of SYP expression in hippocampus by large dose of propofol may explain the mechanism.`
7.Effect of MECT on learning memory in depressed rats and its synaptic plasticity mechanism
Yuanyuan LIU ; Su MIN ; Jun DONG ; Jun CAO ; Ping LI ; Yongfeng LIU
Chinese Journal of Nervous and Mental Diseases 2010;36(2):70-74
Objective To explore the effect of MECT on learning memory in depressed rats and its synaptic plasticity mechanism. Methods Fifty SD rats were randomly divided into 5 groups( n =10): MECT (received ECT with intraperitoneal propofol), ECT (received ECT only), propofol (received intraperitoneal propofol), depression, and control. The treatments were given daily for 7 consecutive days. All rats underwent open field and Morris water maze test. The SYP protein and mRNA expressions in rat hippocampus were detected using immunochemistry and RT-PCR, respectively. Results After treatment, the open field scores were much higher in MECT and ECT groups than in propofol and depression groups ( P <0.05); the learning memory was worse in ECT group than in MECT, propofol and depression groups ( P <0.05); the expressions of SYP protein and mRNA was higher in MECT and ECT groups than in propofol and depression groups ( P <0.05), the expressions of SYP protein and mRNA were lower in MECT group than ECT group ( P <0.05). Conclusions Propofol can improve learning memory in ECT-treated depressed rats through attenuation of ECT-induced expression of SYP in rat hippocampus.
8.The study of cognitive function and synaptic plasticity in depressed rats following electroconvulsive shock
Ping LI ; Xuechao HAO ; Feng LV ; Ke WEI ; Jie LUO ; Jun CAO ; Jun DONG ; Su MIN
Chinese Journal of Nervous and Mental Diseases 2014;(12):715-720
Objective To investigate the role of autophagy and synaptophysin (SYP) in cognitive impairment in de?pressed rats receiving electroconvulsive shock (ECS). Methods Clean and healthy adult male Sprague-Dawley rats were acclimatized to a standard laboratory environment for 7 days. The chronic unpredictable mild stress (CUMS) was used to establish the rat model of depression. Behavior tests were conducted before and after CUMS to evaluate the depression and cognition level of rats. After establishment of the model, 24 rats were randomly divided into ESC group (group E) and depression group (group D) with 12 rats in each group. The rats in group E were administered 80 mg/kg of propofol (10 mg/mL) by intraperitoneal injection, followed by ECS treatment. The rats in group D were administered propofol by intra?peritoneal injection, followed by sham-ECS treatments. The above interventions were conducted daily for 7 consecutive days. After the interventions, rats underwent behavior tests as before. Subsequently, rats were killed and specimens were collected for measurements. Immunohistochemistry was performed to examine autophagy markers such as Beclin 1 and LC3Ⅱand ELISA was used to detect SYP in the hippocampus. Results Group E after ECS significantly increased the percentage of sucrose preference (68.2%±8.7%), rearing times (7.0±1.9), total horizontal distance [(569.5±70.0) cm], es? cape latency [(21.9±5.3)s] and space exploration time [(20.5±3.9)s] compared with group D or group E before ECS. There was no significant difference in these index between groups before ECS or in group E between before and after ECS(P>0.05). Compared with group D, group E had upregulated protein expression levels of Beclin 1 and LC3Ⅱin CA1, CA3, DG as well as the area near the hippocampus and increased SYP contents (P<0.05). Conclusions Cognitive impairment in depression rats following ECS correlates with activated autophagy and increased SYP by ECS.
9.Teaching effect of applying idea of 'integrating world with specialty' in teaching ward-round
Ping LI ; Su MIN ; Jingchuan FAN ; Bo CHENG ; Ke WEI ; Jun CAO
Chinese Journal of Medical Education Research 2012;11(7):722-725
Objective To investigate the teaching effect of applying idea of ‘ integrating world with specialty' in teaching ward - round.Methods Totally 112 undergraduates from department of anesthesiology in Chongqing medical university were randomly divided into 2 groups:group C and group T.The students in group C and group T were received traditional method and teaching ward-round applying idea of ‘integrating world with specialty' respectively.The teching effects were compared.Results Constituent ratio of performance of written test and defence of case analysis in group C and group T were respectively as follow,excellent ( 12%,19%) vs.(31%,37%),good (25%,19% ) vs.(50%,44% ),middle (54%,50% ) vs.( 19%,13% ),bad (9%,12% ) vs.(0,6% ),and there were statistically differences between the two groups.Conclusion Teaching ward-round applying idea of ‘ integrating world with specialty' integrates medical knowledge with social skills and common sense of life,making it easier for medical students to understand and master professional knowledge,as a result of improving the students' ability of problem analysing and solving.