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Three-dimensional (3D) cell culture model is a system that co-culture carriers with 3D structural materials and different types of cells in vitro to simulate the microenvironment in vivo. This novel cell culture model has been proved to be close to the natural system in vivo. In the process of cell attachment, migration, mitosis and apoptosis, it could produce biological reactions different from that of monolayer cell culture. Therefore, it can be used as an ideal model to evaluate the dynamic pharmacological effects of active substances and the metastasis process of cancer cells. This paper compared and analyzed the different characteristics of cell growth and development under two-dimensional (2D) and 3D model culture and introduced the establishment method of 3D cell model. The application progress of 3D cell culture technology in tumor model and intestinal absorption model was summarized. Finally, the application prospect of 3D cell model in the evaluation and screening of active substance was revealed. This review is expected to provide reference for the development and application of new 3D cell culture models.
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Cell Culture Techniques, Three Dimensional , Cell Culture Techniques , Apoptosis , Cell Proliferation , TechnologyABSTRACT
Objective:To treat mice with Alzheimer's disease （AD） with β-catenin RNA interference （RNAi） Huangjingwan （HW）， so as to explore the neuroprotective signal mechanism of its prevention and treatment of AD. Method:A total of 81 male Kunming mice were randomly divided into normal control group， sham model control group， AD model group， Donepezil group， HW+scrambled group， HW+RNAi group， HW group， with 8 mice in each of donepezil group and HW group， and 13 mice in each of other groups. The AD models were established through injection with D-galactose and scopolamine in the last 5 groups for 5 consecutive weeks. On the 1st day of the 4th week after modeling， 0.75 μL PEI-LMW/β-catenin siRNAs nano-complex was injected into the right lateral ventricle of each mouse in for one time to treat with β-catenin RNAi in mice brains of the HW+RNAi group. The 0.75 μL complex was injected into the right lateral ventricle of each mouse for one time as for β-catenin interference control of the HW+scrambled group. The 0.75 μL normal saline was injected into the right lateral ventricle of each mouse in one time of the sham control group. Two weeks after intracerebroventricular injection， β-catenin RNAi was confirmed to be successful， and Donepezil （6.5×10-4 g·kg-1） was intragastrically administered to each mouse of donepezil group. HW （2.5 g·kg-1） was intragastrically administered to each mouse of HW group， HW+RNAi group and HW+scrambled group. Normal saline （0.5 mL·d-1） was intragastrically administered to each mouse of the sham control group. All gastric perfusion lasted for 4 weeks. At the end of gavage， the difference in learning and memory ability of mice was evaluated by platform jumping test. Nissl staining was used to count the number of neurons in s1Tr area of cerebral cortex and CA1 and CA3 areas of hippocampus of each mouse in each group. The mRNA expressions of Wnt1， DVL2， GSK-3β， β-catenin and CyclinD1 in mice brain of each group were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Western blot was used to detect the expressions of Wnt1， DVL2， GSK-3β， β-catenin and CyclinD1 in mice brain of each group. Result:The expression of β-catenin could be significantly inhibited through the injection with PEI-LMW/β-catenin siRNAs nano-complex into the lateral ventricle of AD mice， and nearly no β-catenin expression could be detected， which successfully achieved gene silencing. Compared with the normal control group， mice in AD model group showed that the learning and memory performance decreased significantly， the number of jumping errors increased （P<0.01）， the number of neurons in S1Tr area of cerebral cortex and CA1， CA3 areas of hippocampus decreased significantly （P<0.01）， the mRNA and protein expressions of Wnt1， DVL2， β-catenin， CyclinD1 in brain decreased significantly （P<0.01）， while the mRNA and protein expressions of GSK-3β increased significantly （P<0.01）. Compared with the AD model group， mice in HW group showed that the learning and memory performance increased significantly， the number of jumping errors decreased， the number of neurons in S1Tr area of cerebral cortex and CA1， CA3 areas of hippocampus increased significantly， the mRNA and protein expressions of Wnt1， DVL2， β-catenin， CyclinD1 in brain increased significantly， while the mRNA and protein expression of GSK-3β decreased significantly （P<0.01）. Compared with the HW group， mice in HW+RNAi group showed that the learning and memory performance decreased significantly， the number of jumping errors increased significantly （P<0.01）， the number of neurons in S1Tr area of cerebral cortex and CA1， CA3 areas of hippocampus decreased significantly （P<0.01）， there was no significant change in mRNA and protein expressions of Wnt1， DVL2， GSK-3β in the brain， and the mRNA and protein expressions of β-catenin， CyclinD1 decreased significantly （P<0.01）. Conclusion:HW can treat and prevent AD by activating Wnt/β-catenin signal pathway.
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Alzheimer's disease （AD） is a neurodegenerative disease that leads to progressive memory and cognitive impairment and behavioral disorders， which has seriously threatened the health of the majority of middle-aged and elderly people. Traditional Chinese medicine （TCM） believes that the basic pathogenesis of AD is deficiency of kidney-essence， blood stasis and meridian stagnation. In recent years， many studies have shown that TCM has obvious value and advantages in the prevention and treatment of AD by multi-target mechanism. Therefore， it is of great significance to screen out effective anti-AD drugs from TCM compound prescriptions. Huangjingwan， also known as Jiuzhuan Huangjingwan， has the effects in tonifying kidney-essence， activating blood and removing stasis， with a potential effect in preventing AD. In this article， the feasibility of Huangjingwan in the prevention and treatment of AD was analyzed and discussed from the perspective of TCM theory， the study results of Huangjingwan in the prevention and treatment of AD were summarized， and the mechanism of its action was analyzed from the perspective of pharmacological mechanism. Based on TCM theory， Huangjingwan has the effect of anti-AD. According to relevant findings， Huangjingwan has many targets， such as anti-oxidation， anti-inflammatory， decrease of the level of oxidative stress in brain， activation of Wnt/β-catenin signal transduction in brain， regulation of glycogen synthase kinase-3β （GSK-3β）， protein phosphatase 2A （PP2A） activity balance， reduction of amyloid β （Aβ） content and tau protein hyperphosphorylation in brain， so as to exert effects in improving neurological symptoms and increasing learning and memory ability， with an anti-AD neuroprotective function. This will provide new ideas for in-depth studies and clinical applications of Huangjingwan against AD.
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Objective To determine the optimal positive end-expiratory pressure (PEEP) for volume-controlled ventilation using pulmonary electrical impedance tomography in the patients undergoing surgery with general anesthesia.Methods Fifty patients of both sexes,aged 18-64 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,with body mass index of 18.5-28.0 kg/m2,scheduled for surgery for ureteral calculi under general anesthesia,were enrolled in this study.The patients were tracheally intubated after anesthesia induction and mechanically ventilated in volume-controlled mode,with tidal volume 6 ml/kg,mean arterial pressure was recorded at 3 min of ventilation and served as the baseline value,and then PEEP was increased with an increment of 3 cmH2O every 3 min until PEEP reached 15 cmH2 O.The percentage of dorsal pulmonary ventilation and peak airway pressure were recorded at 3 min of ventilation with different PEEPs.When the decrease in mean arterial pressure was more than 20％ of the baseline value during ventilation,deoxyepinephrine 0.1 mg was injected intravenously,and the consumption of deoxyepinephrine was recorded within 3 min of ventilation with different PEEPs.Results Peak airway pressure was gradually increased with the increase of PEEP (P＜0.05),the percentage of dorsal pulmonary ventilation was gradually increased when PEEP was 6 cmH2O (P＜ 0.05),and the consumption of deoxyepinephrine was gradually increased when PEEP was 15 cmH2O (P＜0.05).Conclusion The optimal PEEP is 12 cmH2O during volume-controlled ventilation with tidal volume of 6 ml/kg in the patients undergoing surgery with general anesthesia.
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OBJECTIVE: To observe the effect of electroacupuncture (EA) at different time-points on the injured myocar-dium and expression of myocardial Bax/Bcl-2 and Lc 3 Ⅱ/Ⅰ proteins in acute myocardial ischemia-reperfusion injury (MIRI) rats so as to explore its mechanisms underlying myocardial protective effect via reducing cardiomyocyte autophagy and apoptosis. METHODS: A total of 66 adult SD rats were randomly divided into sham operation (sham), model, EA-R 0min(R＝ reperfusion), EA-R 30min, EA-R 60min, and EA-R 120min groups, with 6 rats being in the sham group and 12 rats being in each of the other 5 groups. The MIRI model was prepared by ligating the anterior descending branch (ADB) of the left coronary artery for 30 min followed by reperfusion for 4 h. In the sham group, the ADB was only threaded without ligation. EA was applied to bilateral "Neiguan" (PC 6) for 20 min at 0, 30, 60, and 120 min when reperfusion. Evans Blue-triphenyltetrazolium chloride (TTC) double staining was performed to determine the myocardial infarction area (MIA) and the ratio of the infarct size of the area at risk (IS/AAR). ELISA was performed to measure serum cardiac troponin 1 (cTn-Ⅰ) content, and Western blot was used to measure the expression of apoptosis-related proteins Bax and Bcl-2 and autophagy related proteins Lc 3 Ⅱ and Lc 3 Ⅰ in the left cardiac ventricle tissue. RESULTS: Compared with the model group, the percentages of MIA in the EA-R 30min, EA-R 60min, and EA-R 120min groups, and the IS/AAR in the EA-R 0min, EA-R 30min, EA-R 60min and EA-R 120min groups were significantly reduced (P0.05). CONCLUSION: EA intervention can reduce MIA in MIRI rats, which is possibly closely related to its effects in reducing apoptosis and autophagy. The best intervention time is at 30 min after MI reperfusion, but the difference of effects of EA at different time-points is independent of Bax/Bcl-2 and Lc 3 Ⅱ/Ⅰ expression.
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Objective To investigate the clinical efficacy of minimally invasive esophagectomy and open triple-incision esophagectomy for esophageal cancer (EC).Methods The retrospective cohort study was conducted.The clinicopathological data of 454 EC patients who were admitted to the Tianjin Medical University Cancer Institute and Hospital from January 2012 to September 2016 were collected.Of 454 patients,229 undergoing thoracoscopic esophagectomy (194) or combined thoracoscopic + laparoscopic esophagectomy (35) were allocated into the minimally invasive group,and 225 undergoing open triple-incision esophagectomy in the left cervical,right chest and epigastric regions were allocated into the open group.Observation indicators:(1) intraoperative situations;(2) postoperative recovery situations;(3) stratified analysis;(4) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect the postoperative survival up to October 2017.Measurement data with normal distribution were represented as-x±s,and t test was used for comparison between groups.Measurement data with skewed distribution were described as M (range),non-parametric test was used for comparison between groups.Count data were expressed as percentage,and the chi-square test or fisher exact probability method were used to test comparison between groups.KaplanMeier method was used to calculate survival rate and draw survival curve.Log-rank test was used for survival analysis.Results (1) Intraoperative situations:operation time,numbers of upper mediastina lymph node dissected and right laryngeal nerve lymph node dissected in stage 0-Ⅱ of TNM staging and numbers of neck lymph nodes dissected in stage Ⅲ of TNM staging were respectively (307±70)minutes,4 (range,0-18),2 (range,0-10),0 (range,0-24) in the minimally invasive group and (267±49)minutes,3 (range,0-15),1 (range,0-7),0 (range,0-46) in the open group,with statistically significant differences between groups (t =7.071,Z=-2.207,-2.717,-1.969,P＜0.05).(2) Postoperative recovery situations:thoracic drainage-tube removal time and volume of drainage fluid were respectively 5 days (range,2-88 days),280 mL (range,0-7 792 mL)in the minimally invasive group and 8 days (range,1-72 days),1 650 mL (range,225-7 970 mL),with statistically significant differences between groups (Z =-9.618,-15.443,P ＜ 0.05).The cases with total postoperative complications,arrhythmia and recurrent laryngeal nerve paralysis were 72,20,35 in the minimally invasive group and 100,36,56 in the open group,with statistically significant differences between groups (x2=8.155,5.542,6.533,P＜0.05).Patients may be combined with multiple complications.Two patients died within 30 days postoperatively,including 1 with respiratory failure and 1 with pulmonary embolism.Patients with other complications were improved after symptomatic and supportive treatments.(3) Stratified analysis:of 229 patients in the minimally invasive group,93 underwent surgery within the physician's learning curve and 136 underwent surgery after physician's learning curve.Operation time,volume of intraoperative blood loss,dissected numbers of upper mediastina lymph node,right laryngeal nerve lymph node,left laryngeal nerve lymph node,middle mediastinal lymph node and lower mediastinal lymph node,cases with pneumonia,recurrent laryngeal nerve paralysis,chylothorax,anastomotic stenosis,anastomotic fistula,respiratory failure and pulmonary embolism in 93 patients were respectively (306±68)minutes,(217± 178)mL,3 (range,0-20),2 (range,0-8),0 (range,0-10),6(range,0-17),1 (range,0-6),5,16,1,5,3,2,2 in the minimally invasive group and (308±72)minutes,(200±112)mL,4 (range,0-37),2 (range,0-10),0 (range,0-8),7 (range,0-20),1 (range,0-10),4,19,3,3,4,4,0 in the open group,with a statistically significant difference in number of upper mediastina lymph node dissected between groups (Z=-2.472,P＜0.05) and no statistically significant difference in other indicators between groups (t =-0.160,0.917,Z =-0.113,-1.698,-0.950,-0.510,x2 =0.342,0.446,P＞0.05).(4) Follow-up and survival situations:of 454 patients,415 were followed up for 1-62 months,with a median time of 28 months.Among the 415 patients,operation time ≥ 3 years was detected in 162 patients,(77 in the minimally invasive group and 85 in the open group),and 3-year cumulative survival rates of the minimally invasive and open groups were 68.1％ and 53.8％,showing no statistically significant difference between groups (x2=3.293,P＞0.05).Further subgroup analysis showed that postoperative 3-year cumulative survival rates of patients with the stage Ⅰ-Ⅱ and Ⅲ of TNM staging were respectively 82.1％,53.7％ in the minimally invasive group and 62.6％,48.6％ in the open group,showing no statistically significant difference between groups (x2=2.664,0.382,P＞ 0.05).Conclusion Minimally invasive esophagectomy has some characteristics of less surgical trauma postoperative complications,and its resection effect is comparable to open esophagectomy.
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Objective@#To compare and evaluate the prognostic value of the 7th and 8th edition of The AJCC Esophageal Cancer Staging System for patients with stage Ⅱ and Ⅲ esophageal squamous cell carcinoma.@*Methods@#The clinical data of 328 esophageal cancer patients who received operation at Department of Esophageal Cancer, Tianjin Tumour Hospital from January 2006 to December 2010 were restrospectively analyzed. There were 63 female and 265 male patients. The mean age was 65 (range: 33 to 87) years. Univariate and multivariate analysis were performed to identify the prognosis factors.@*Results@#The five years overall survival rates among patients with stage Ⅱ and Ⅲ were both significantly different (χ2=87.035, 84.730, all P=0.000) according to the 7th and 8th editions of the TNM staging systems. The five years overall survival rate among patients with stage ⅡB and ⅢA were significantly different (39.6% vs 23.4%, P=0.001) according to the 7th edition of the esophageal cancer staging systems.According to the 8th edition of the esophageal cancer staging system, the 5 years survival rate of patients with stage ⅡA and ⅡB, ⅢB and Ⅳ was statistically significant (58.5% vs. 35.5%, P=0.040; 18.9% vs. 0, P=0.000). In multivariate analysis, tumor size, T staging, N staging and tumor differentiation (HR=1.592, 95%CI: 1.185 to 2.139, P=0.002; HR=1.519, 95% CI: 1.236 to 1.867, P=0.000; HR=1.647, 95% CI: 1.448 to 1.874, P=0.000; HR=1.404, 95% CI: 1.059 to 1.861, P=0.018) were the main independent prognosis factors affecting the prognosis of esophageal squamous cell carcinoma patients.@*Conclusions@#Both the 7th and the 8th editions of TNM staging systems are able to reflect the clinical prognosis of patients receiving radical resection of esophageal cancer, and the factors of tumor size, differentiaton, invasion depth and lymph node metastases are the independent predictors of prognosis. The 8th edition provides a more detailed and more reasonable for the staging of stage Ⅱ and Ⅲ for esophageal cancer patients than the 7th edition, and it is more accurate for the prognosis of patients with esophageal cancer after surgery.
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Objective To investigate the effect of mild hypothermia combined with mitochondrial divison inhibitor 1 in mitochondrial after cerebral ischemia-reperfusion (IR).Methods Fourty male healthy Sprague-Dawley (SD) rats, weighing 280-320 g, were randomly divided into 5 groups (n=8 each): group Sham, group IR, hypothermia group (group H), Mdivi-1 group (group M) and hypothermia+Mdivi-1 group (group HM).Animal models of global cerebral IR were established by transoesophageal cardiac pacing inducing cardiac arrest followed by cardiopulmonary resuscitation (ischemia 4 min and reperfusion 6 h).The group Sham was similarly treated to group IR except the cardiac arrest and cardiopulmonary resuscitation.In groups H and HM, the core temperature was cooled down to 32-34℃ within 15 min starting from the beginning of reperfusion, and maintained for 6 h.In the other groups, the core temperature was maintained at the normal temperature.In groups M and HM, the animals were given Mdivi-1 (1.2 mg/kg) intravenously at the beginning of the reperfusion and the other groups were given the same Volume of dimethylsnlfone (DMSO).After 6 h of reperfusion, the rats were sacrificed, and bilateral hippocampi were immediately removed for determination the protein level of dynamin-related proten 1 (Drp1) and cytochrome C (Cyt-C) expression by Western blot and obsevation of the mitochondrial structure of pyramidal cell in hippocampal CA1 under electronic microscope.Results Compared with group Sham, the expression of Drp1 and Cyt-C was up-regulated in groups IR, H, M and HM (P＜0.05).Compared with group IR, the expression of Drp1 and Cyt-C was down-regulated in groups H, M and HM (P＜0.05).Compared with groups H and M, the expression of Drp1 and Cyt-C was down-regulated in group HM (P＜0.05).There was no significant difference in the expression of Drp1 and Cyt-C between groups H and M.The mitochondria were rod-shaped with clear and sound structure in group Sham, while mitochondria showed various degree of fission, swollen structures, matrix deposit, vacuoles formation and cristae collapse in other groups.The changes of group HM were relatively slight.Conclusion Mild hypothermia combined with mitochondrial divison inhibitor 1 alleviate mitochondrial damage after global cerebral IR of rats.The combined effect is better than that of any individual application.
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In recent years,Shanghai is exploring the implementation ot comprehensive budget management in community nealtn center,but facing some problems in process:the budget management model is still not unified,"separation between revenue and expenditure" policy is not really implemented,the financial subsidies is not combined with performance appraisal.Meanwhile within the center,the personnel funds are approved by public sector,the operation funds are not standardized,and project funds are relatively chaotic.It is recommended to carry out cost accounting of community health center,clarify expenditure,introduce standardized workload in community health center to promote budgeting scientifically,strengthen the information construction to make budget implementation assessment,and establish a performance appraisal linked to the government subsidy mechanism.
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Objective To evaluate the effect of hydrogen sulfide on hippocampal endoplasmic reticulum stress during global cerebral ischemia-reperfusion (I/R) in rats.Methods Seventy-two pathogen-free healthy male Sprague-Dawley rats,weighing 280-320 g,aged 8-10 weeks,were divided into 3 groups (n=24 each) using a random number table:sham operation group (group Sham),global cerebral I/R group (group I/R) and global cerebral I/R plus sodium hydrosulfide group (group I/R+NaHS).Cardiac arrest was induced with transoesophageal cardiac pacing followed by cardiopulmonary resuscitation to establish the global cerebral I/R model.Immediately after recovery of spontaneous circulation,sodium hydrosulfide 2.5 mg/kg was intraperitoneally injected in group I/R+NaHS,and normal saline 5 ml/kg was given in group I/R.The hippocampi were immediately removed at 24 h of reperfusion for determination of the expression of glucose-regulated protein 78 (GRP78),C/EBP-homologous protein (CHOP) and caspase-12 in hippocampal tissues (by Western blot).At 1,3 and 7 days of reperfusion,the hippocampal tissues were obtained and stained with haematoxylin and eosin for examination of the pathological changes in hippocampal CA1 region (under a light microscope) and for determination of apoptosis in hippocampal cells (using TUNEL staining),and the apoptosis rate was calculated.Results Compared with group Sham,the apoptosis rate of hippocampal tissues at 1,3 and 7 days of reperfusion in group I/R and at 3 and 7 days of reperfusion in group I/R+NaHS were significantly increased,and the expression of GRP78,CHOP and caspase-12 in hippocampal tissues was significantly up-regulated in I/R and I/R+NaHS groups (P＜0.05).Compared with group I/R,the apoptosis rate of hippocampal tissues was significantly decreased,and the expression of GRP78,CHOP and caspase-12 was down-regulated at 1,3 and 7 days of reperfusion (P＜0.05),and the pathological changes were significantly attenuated in group I/R+NaHS.Conclusion The mechanism by which hydrogen sulfide reduces apoptosis in hippocampal cells is related to inhibition of endoplasmic reticulum stress during global cerebral I/R in rats.
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The industry of germ-free animals has been a hot spot in research along with the rapid development of studies on the relationship between microbiota and host diseases. Because it is pathogen?free, and the high degree of simi?larity in anatomy, physiology, pathogenesis to humans, germ?free pig is considered a clinical relevant model to be widely used in life science research. Based on the current state of research of germ?free pig cultivation at home and abroad and the experimental studies carried out in our laboratory as well, this article gives a simple discussion on germ?free technique of domestic pigs.
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OBJECTIVE: To establish the cell model of human neuroblastoma cell( SH-SY5Y cell) exposed to1,2-dichloroethane( 1,2-DCE) in vitro and to explore the mechanism of 1,2-DCE-induced toxicity in SH-SY5Y cells.METHODS: SH-SY5Y cells were collected in their logarithmic growth phase and cultured in complete medium that had final concentrations of 1,2-DCE in 0,10,20,30,40,50,60,70 and 80 mmol / L for 24 hours. Cell morphology was observed and cell survival rate was examined by CCK-8 assay. Using chemical colorimetric method, the activity of lactic dehydrogenase( LDH) in the cell culture supernatant,and the intracellular level of malondialdehyde( MDA),the intracellular activities of superoxide dismutase( SOD) and adenosine triphosphate( ATP) enzymes were detected. RESULTS: With the increasing exposure concentrations of 1,2-DCE,the cell density of SH-SY5Y cells gradually decreased,the synapse became shorter,the membrane ruptured,cytoplasm condensed and cytoplasmic contents overflowed increased.With the increasing concentration of 1,2-DCE,the cell survival rate decreased( P < 0. 01),the activity of LDH in the cell culture supernatant increased( P < 0. 01). These changes had a dose-effect correlation. Intracellular MDA level,and activities of SOD,Na~+-K~+-ATP enzyme,Ca~(2+)-Mg~(2+)-ATP enzyme and total ATP enzyme increased at first and then decreased. The activity of LDH in the cell culture supernatant and cell survival rate was negatively correlated( the correlation coefficient is- 0. 907,P < 0. 01). CONCLUSION: 1,2-DCE could inhibit the proliferation of SH-SY5Y cells.The mechanism may be related to the permeability change of cell membrane,cellular damage from excessive free radicals,the decrease of free radical scavenging capacity,ATP enzyme activity and calcium overloading. SH-SY5Y cells can be used as a common cell line for 1,2-DCE cytotoxicity analysis.
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OBJECTIVE: To determine the effects of 1-bromopropane( 1-BP) subacute inhalation on the expression of neuron specific enolase( NSE) and myelin basic protein( MBP) in plasma and brain tissue in male rats. METHODS: Specific pathogen free adult male Wistar rats were randomly divided into 4 groups with 12 rats in each group: the control group,the low-,medium- and high-dose groups with 1-BP exposure levels at 0,1 250,2 500 and 5 000 mg / m3,respectively. The rats were given continuous dynamic inhalation of 1-BP for 6 hours per day,5 days per week,for continuous 4 weeks. The rats were sacrificed at the end of exposure,9 rats from each group were randomly chosen and the blood from abdominal aorta was collected and the plasma was isolated. The plasma levels of NSE and MBP were measured by enzyme-linked immunosorbent assay. The whole brain,pallium,cerebellum and brainstem were isolated for detection of organ coefficient.The rest of 3 rats in each group were processed for histopathologic examination and the expressions of NSE and MBP were evaluated by immunohistochemistry. RESULTS: The organ coefficients of whole brain,pallium,cerebellum and brainstem in the high-dose group were higher than those in the control group [( 0. 754 ± 0. 056) % vs( 0. 663 ± 0. 035) %,( 0. 382 ±0. 037) % vs( 0. 339 ± 0. 021) %,( 0. 115 ± 0. 008) % vs( 0. 098 ± 0. 006) % and( 0. 213 ± 0. 018) % vs( 0. 183 ±0. 014) %,respectively,P < 0. 01]. The plasma NSE levels in the 3 exposure groups were lower than those of control group [( 7. 92 ± 0. 53) vs( 24. 73 ± 11. 44),( 9. 12 ± 2. 17) vs( 24. 73 ± 11. 44) and( 11. 10 ± 2. 84) vs( 24. 73 ±11. 44) mg / L,respectively,P < 0. 01]. The plasma MBP levels in all groups showed no statistical difference [( 2. 52 ±0. 70) vs( 2. 50 ± 0. 72) vs( 2. 47 ± 0. 66) vs( 2. 44 ± 0. 81) mg / L,P > 0. 05]. Histopathological examination showed that a few necrotic nerve cells were observed in hippocampus of rats in high-dose group. The expressions of NSE and MBP in brain tissue of rats in control,low- and medium-dose groups showed no significant difference. The down-regulated expression of NSE and MBP were only observed in cells of hippocampus of rats in the high-dose group. CONCLUSION: The1-BP induced neural toxicity was reflected in the function of central nervous system rather than in the structural morphology. The plasma NSE might be one of the effect biomarkers for monitoring 1-BP exposure.
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OBJECTIVE: To study the potential effects of subacute 1-bromopropane( 1-BP) inhalation on the expression of synapse specific biomarkers synaptophysin( SYP),glutamate receptor 2( GluR2) and N-methyl-D-aspartate receptor 2B( NR2B) in the hippocampus of male rats. METHODS: Forty-eight specific pathogen free adult male Wistar rats were randomly divided into control group,low-,medium-,and high-dose groups according to body weight. Each group consisted of 12 rats. By dynamic inhalation intoxication method,the control group was exposed to fresh air,the dose groups were given 1 250,2 500 and 5 000 mg / m3 of 1-BP respectively,6 hours per day,5 days per week for continuous 4 weeks. After the exposure,the rats were executed and the whole brains were separated into cerebrum( included hippocampus),brainstem and cerebellum. Real time quantitative polymerase chain reaction and Western blot were used for detection of SYP,GluR2 and NR2 B mRNA and protein expression in hippocampus. RESULTS: Slow response and muscle strength descended in hind limbs were found in high-dose group in the 3rd week. Body weights of rats in high-dose group were lower than those of control group from the 1st to the 4th week( P < 0. 01). Weights of whole brain,cerebrum and brainstem in high-dose group were lower than those of control group( P < 0. 05). Rats in high-does group were found neuron necrosis in hippocampus cornu ammonis 3 and dentate gyrus region. No significant difference was found in SYP,GluR2 and NR2 B mRNA relative expression in all groups( P > 0. 05). No significant difference was found in SYP protein relative expression in different groups( P > 0. 05). The GluR2 protein relative expression in high-dose group was lower than that of control group( P < 0. 05). The NR2 B protein relative expression was higher than that of control group( P < 0. 05). CONCLUSION: The GluR2 and NR2 B protein expression in hippocampus can be potential biomarkers for 1-BP central neurotoxicity,but its physiological meaning needs further elucidation.
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Objective To evaluate the effect of hypothermia on the expression of dynamin?related protein 1 ( Drp1) in brain tissues during global cerebral ischemia?reperfusion ( I∕R) in rats. Methods Thirty?six healthy male Sprague?Dawley rats, weighing 280-320 g, were divided into 3 groups ( n=12 each) using a random number table: sham operation group ( group Sham ) , global cerebral I∕R group ( group I∕R) and hypothermia group ( group H) . Cardiac arrest was induced by transoesophageal cardiac pacing followed by cardiopulmonary resuscitation to establish the global cerebral I∕R model in anesthetized rats in I∕R and H groups. In group H, the body temperature ( rectal temperature) was cooled down to 32-34 ℃ within 15 min starting from the beginning of reperfusion, and maintained at this level for 6 h. At 72 h of reperfusion, neurological deficit was scored, and the rats were sacrificed, and the whole brain was removed for examination of the pathological changes in hippocampal CA1 region and for determination of nor?mal pyramidal cell count and neuronal apoptosis in hippocampal CA1 region and expression of Drp1 and cy?tochrome c (Cyt c) in hippocampal tissues (by Western blot). The apoptosis rate was calculated. Re?sults Compared with group S, the neurological deficit score and apoptosis rate were significantly in?creased, and the number of normal pyramidal cells was decreased in I∕R and H groups, the expression of Drp1 and Cyt c in hippocampal tissues was significantly up?regulated in group I∕R ( P0.05) . Compared with group I∕R, the neurological deficit score and apoptosis rate were significantly de?creased, the number of normal pyramidal cells was increased, and the expression of Drp1 and Cyt c in hip?pocampal tissues was down?regulated in group H ( P<0.05) . Conclusion The mechanism by which hypo?thermia inhibits cell apoptosis during global cerebral I∕R may be related to down?regulation of Drp1 expres?sion in rats.
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Purpose To analyze the relationships between glomerular filtration rate (GFR) measured by SPECT and renal parenchyma thickness and enhanced CT value measured by CT, and to explore predictive significance of CT in evaluating renal function of patients with hydronephrosis.Materials and Methods One hundred and fifteen patients diagnosed with unilateral hydronephrosis by ultrasound were retrospectively analyzed. GFR% (GFR percentage of affected kidney to the both kidneys) was measured by SPECT. CT% (percentage of affected renal parenchyma thickness to the both kidneys) and enhanced CT% (percentage of enhanced CT value of affected kidney to the both kidneys) were measured by pre- and post-contrast CT scan. According to GFR, the renal function was divided into mild-to-moderate impairment, severely impairment and non-function. Twenty-five volunteers were recruited as control group. CT%, enhanced CT% and GFR%among the four groups were compared, and the correlation of CT% and enhanced CT%with GFR% was analyzed to evaluated CT in predicting renal function.Results CT%, enhanced CT% and GFR% in mild-to-moderate impairment group was significantly greater than those in severely impairment group and non function group (F=20.24, 7.78 and 329.21,P<0.05). GFR% was positive correlated with CT% (r=0.58,P<0.05) and enhanced CT% (r=0.61,P<0.05). Area under curve (AUC) of CT% were 0.54, 0.79 and 0.83 for mild-to-moderate impairment, severely impairment and non-function, with sensitivity of 92.91%, 93.47%, 65.72%, and specificity of 35.33%, 59.47%, and 88.62%, respectively. AUC of enhanced CT% were 0.79, 0.89 and 0.96 for the three groups, with sensitivity of 97.51%, 80.02%, 97.66%, and specificity of 58.14%, 89.82% and 94.27%, respectively.Conclusion There was high correlation between renal function imaging by SPECT and CT in evaluating renal function of hydroneohrosis patients. Pre- and post-contrast CT scan can be used as complements in predicting renal function, and post-contrast CT with high accuracy.
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To establish a method for the determination of cucurbitacin in plasma samples, in order to study the in vivo pharmacokinetic characteristics of cucurbitacin in rats. Rats were intravenously injected with cucurbitacin. With diphenhydramine as the internal standard (IS), the plasma concentrations of cucurbitacin in rat plasma at different time points were determined by liquid chromatography tandem mass spectrometry (LC-MS/MS). With electrospray ionization source, the positive ion detection in the multiple reaction monitoring mode was conducted to determine the ion-pairs for target compound and IS were m/z 503.2/113.1 and m/z 256.0/167.2, respectively. Agilent ZOBAX SB-C18 column (2.1 mm x 50 mm, 1.8 microm) was adopted and eluted with methanol and 0.1% formic acid (55:45), and the flow rate was 0.2 mL x min(-1). DAS 2.0 software was applied to fit the blood concentration and calculate corresponding pharmacokinetic parameters. The rats were intravenously injected with cucurbitacin at the concentration of 3.0 mg x kg(-1). The target blood quality concentration show good linear relations within the range of 10.5-3 150 microg x L(-1) (R2 = 0.996), the lower limit of the standard curve was 10.5 microg x L(-1), and the signal to noise ratio S/N = 12. Intra- and inter-day precisions RSD was less than 6.9% and 14%, respectively; The accuracy RE ranged between 0.20% and 3.7%; The extraction recoveries ranged between 92.7% and 97.1%. Regarding the pharmacokinetic parameters of tail intravenous injection of cucurbitacin, AUC (0-t) was (811.615 +/- 111.578) microg x h x L(-1), (t1/2) was (1.285 +/- 1.390) h, CL was (3.627 +/- 0.487) L x h x kg(-1), and V(d) was (6.721 +/- 7.429) L x kg(-1). In this study, researchers established a simple, accurate, sensitive and highly specific method for determining the blood concentration of cucurbitacin, and reported the in vivo pharmacokinetic characteristics of cucurbitacin in rats for the first time.
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Animals , Male , Rats , Administration, Oral , Cucurbitaceae , Chemistry , Cucurbitacins , Blood , Pharmacokinetics , Drugs, Chinese Herbal , Pharmacokinetics , Rats, WistarABSTRACT
To compare the pharmacokinetics of syringin, eleutheroside E and isofraxidin after intravenous administration of each monomer and Ciwujia injection. Twenty-four Sprague-Dawley rats were randomly divided into four groups and intravenously administrated with syringin, eleutheroside E, isofraxidin, and Ciwujia injection, respectively. The concentrations of the three components in rat plasma were determined by LC-MS/MS. DAS 2.0 software was applied to calculate the pharmacokinetic parameters while the SPSS 17.0 software was used for statistical analysis. Significant difference (P < 0.05) was found between each monomer and the injection on the main pharmacokinetic parameters such as AUC, CL and t1,/2. Compared with the injection, the group treated with the syringin has obvious decrease in AUC, and increase in CL while the group treated with eleutheroside E has obvious increase in AUC, and decrease in CL The t1/2 of isofraxidin was prolonged in Ciwujia injection. Pharmacokinetic characters of the ingredients in the injection varied greatly from the monomer. Other constituents in the injection may have an impact on the pharmacokinetic profiles of these three components.
Subject(s)
Animals , Male , Rats , Administration, Intravenous , Coumarins , Blood , Pharmacokinetics , Drugs, Chinese Herbal , Pharmacokinetics , Glucosides , Blood , Pharmacokinetics , Lignans , Blood , Pharmacokinetics , Phenylpropionates , Blood , Pharmacokinetics , Rats, Sprague-DawleyABSTRACT
Objective:To establish a GC-MS method for the simultaneous determination of 16 phthalate ester plasticizers in oral liquid preparations, compare the purification effect of liquid preparations by liquid-liquid extraction and solid phase extraction and opti-mize the operating parameters to determine the optimal experimental conditions. Methods:The samples were operated by liquid-liquid extraction and solid phase extraction, respectively. Selective ion monitor ( SIM) was adopt, phthalate esters were identified by the rela-tive abundance of major characteristic ions and the content was determined by an external standard method. Results:When the samples were operated by liquid-liquid extraction, the interference was strong with many impurity peaks. Therefore, the solid phase extraction was adopted for the sample pretreatment. GC-MS was used to detect the residues of 16 phthalate ester plasticizers in oral liquid prepara-tions. The detection limit was 0. 02μg·g-1 ,while the calibration curve showed good linearity within the range of 0. 25-8. 0μg·ml-1 with the correlation coefficient between 0. 990 7 and 0. 999 8. The average recoveries were 76. 0%-95. 4%. The relative standard devi-ations were between 2. 3% and 9. 6%(n=6). Conclusion: Pretreated by solid phase extraction, the residues of 16 phthalate ester plasticizers in oral liquid preparations can be detected by GC-MS with the properties of simple, fast, precise and sensitive, and it is suitable for the determination of phthalate esters in oral liquid preparations.
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<p><b>OBJECTIVE</b>To develop an effective identification method for accurately discriminating Psammosilene tunicoides and its confused species by the combined method of microscopic identification and molecular identification, so-called systematic identification of Chinese materia medica (SICMM).</p><p><b>METHOD</b>P. tunicoides and its confused species were accurately discriminated by SICMM method, which was established by comprehensively use of microscopic identification and DNA identification method. The DNA identification included the following analysis: the BLAST alignment, specific bases and N-J phylogenetic tree analysis.</p><p><b>RESULT</b>The cluster crystals were not observed in P. tunicoides, but great deals of them were found in Silene viscidula. Further more, big differences of ITS sequence were observed and analyzed between P. tunicoides and its confused specie of S. viscidula.</p><p><b>CONCLUSION</b>The system method is a scientific and accurate method for the identification of P. tunicoides and its counterfeit species.</p>