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Objective To observe the effects of Yiqi Jiedu prescription drug serum on the apoptosis of human renal cell carcinoma ACHN cells and Notch1 gene in Notch signal pathway. Methods The healthy SD female rats were selected, which were divided into 4 groups, 8 rats in each group: the rats were fed with the normal saline 10 ml·kg-1·d-1 (the blank serum group); the rats were fed with the Yiqi Jiedu prescription decoctum of 50 g·kg-1·d-1 (the high-concentration of Yiqi Jiedu prescription serum group); the rats were fed with the Yiqi Jiedu prescription decoctum of 25 g·kg -1·d -1 (the medium-concentration of Yiqi Jiedu prescription serum group); the rats were fed with the Yiqi Jiedu prescription decoctum of 12.5 g·kg-1·d-1 (the low-concentration medicated of Yiqi Jiedu prescription serum group); then the serum would be extracted 10 days later in each group. ACHN cells at exponential phase were cultured in the above 4 groups. Theproliferation of ACHN cells in each group was observed by using CCK-8 method. The apoptosis of ACHN cells was detected by using flow cytometry (FCM). The expression levels of Notch1 mRNA of ACHN cells in each groups were detected by using real time fluorescence quantitative polymerase chain reaction (RT-PCR). Results The inhibition rates of ACHN cells in the high-concentration group, the medium-concentration group, the low-concentration group of Yiqi Jiedu prescription and the blank serum group 24 h later were (12.34±4.25)％, (7.47±1.40)％, (2.52±0.62)％, (1.05±0.31)％, respectively (F= 15.04, P< 0.01); after 48 h, the inhibition rates were (24.20 ±2.41)％, (14.23 ±1.56)％, (5.08 ±1.34)％, (1.16 ±0.14)％, respectively (F=227.36, P<0.01);after 72 h, the inhibition rates were (32.16±2.45)％, (25.05±3.69)％, (10.29±2.76)％, (1.07± 0.71)％, respectively (F=110.51, P<0.01). The results showed that Yiqi Jiedu prescription drug serum could significantly inhibit the proliferation of ACHN cells in a dose-dependent manner. Besides, the inhibition rate differences at all time points of the high-concentration serum group (F= 31.44, P< 0.01), the medium-concentration serum group (F= 49.61, P< 0.01) and the low-concentration serum group (F= 68.78, P<0.01) were all statistically significant, and they were in a time-dependent manner. The apoptosis rate of cells in the high-concentration group, the medium-concentration group, the low-concentration group of Yiqi Jiedu prescription and the control serum group was (34.5±1.5)％, (24.4±1.5)％and (13.1±0.5)％, (5.2±0.3)％, respectively, and the differences were statistically significant (F = 1153.36, P < 0.01). The relative expression level of Notch1 mRNA of cells in the high-concentration group, the medium-concentration group, the low-concentration serum group of Yiqi Jiedu prescription and the control serum group was 0.213 ±0.032, 0.432 ±0.049, 0.781 ±0.018, 1.013 ±0.047, respectively, and the differences were statistically significant (F=270.60, P<0.01). Conclusion Yiqi Jiedu prescription drug serum can induce apoptosis of ACHN cells, and its mechanism may be related to the down-regulation of the expression level of Notch1 receptors.
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Objective To isolate and identify stem-like cells (CSCs) from human ACHN renal cell carcinoma cell line.Methods ACHN cancer stem cells CSCs were enriched by serum-free culture condition.Methyl thiazolyl tetrazolium (MTT) method was performed to draw the growth curve.Flow cytometry was performed to evaluate the expressions of two CSCs markers,CD133 and CD44.Real-time polymerase chain reaction (RT-PCR) was performed to evaluate the expression of stem-related genes (Oct3/4,Bmi,Nanog,and 3-catenin).The tumorigenicity in vivo was also examined.Results ACHN CSCs were formed by sphere-forming culturing.The data demonstrated that there were stronger capacities of proliferation in ACHN tumor spheres than that of ACHN cells.And the expressions of CD133 and CD44 were also significandy enhanced in tumor spheres.The data showed that four genes (Oct3/4,Bmi,Nanog,and β-catenin)were upregulated in sphere-forming cells.Sphere-forming cells had a higher tumorigenic potential in vivo than incubated cells.Conclusions Human ACHN renal cell carcinoma cell line sphere-forming cells can be enriched by serum-free culture condition and exhibits several characteristic cancer stem cells properties.
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Objective:To clarify the essence of hospital equipment management and propose the task and development trend of information management need to be accomplished, and solve the bottleneck problem of system design, which is always focused on the process of hospital equipment information management promotion in our country.Methods: Comprehensively analyzed daily work of hospital equipment management, and designed a new dual core equipment information system architecture, which was ex purchasing + post purchasing dual core architecture.Results: Based on the new architecture, the hospital equipment information system can cover the entire equipment management workflow, and it made information transmission between jobs more efficiently, cooperation more closely, responsibilities more clearly, constraint more effectively.Conclusion:The new system architecture emphasizes the role of collective labor. It depends on all the members of the equipment department to work together for the goal of achieving dynamic and full life-cycle equipment management.
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AIM:To observe the Toll-like receptor 9 (TLR9) activation in microglia BV-2 cells after oxygen-glucose deprivation and reoxygenation ( OGDR) , and its effects on neuronal apoptosis.METHODS:The BV-2 cell super-natants were collected after the corresponding treatment and added to mouse primary cortical neurons after OGDR for 4 h, followed by normal culture for 24 h.The cells were divided into normal BV-2 group, NC-siRNA group, TLR9-siRNA group, OGDR group, OGDR+NC-siRNA group, OGDR+TLR9-siRNA group and control group (without adding BV-2 cell supernatant) .The changes of the neuronal morphology were observed under an inverted phase-contrast microscope, and the neuronal apoptosis was detected by TUNEL.The protein expression of cleaved caspase-3 was detected by Western blot-ting.RESULTS:After OGDR, the axon turned thin, twisted and broken, and neuronal swelling, decrease in refraction and vacuolar degeneration were observed.The green-stained apoptotic bodies in the neurons in all groups were positive. Compared with control group, the caspase-3 protein levels in other groups were increased.Compared with the normal BV-2 group, the caspase-3 protein in OGDR group and TLR9-siRNA group was increased.Compared with OGDR+TLR9-siRNA group, the caspase-3 protein in TLR9-siRNA group and OGDR group was decreased.CONCLUSION: After OGDR, TLR9 activation in BV-2 cells induces neuronal apoptosis with the increase in caspase-3 protein level.Inhibition of TLR9 expression reduces neuronal damage.
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Objective To identify regular roots for medical disputes by case studies for providing evidences for hospital management..Methods 200 cases of medical malpractice were selected consecutively and categorized according to the causes,dispute focus and expert opinions.Results The main factor to trigger medical disputes is dissatisfaction of the outcome,accounting for 50.5％.48.15％ of the disputes,however,result from dissatisfaction of the therapy process which constitutes a medical malpractice.Neglect or defects in treatment,surgical operation,information notice and medical papers were highly common causes.Conclusion Medical disputes result from a variety of causes,so are the roots of medical malpractice.Medical institutions are recommended to enhance medical quality management,make sufficient doctor-patient communication,and make high quality documentation of key medical activities,in an effort to minimize medical disputes and medical malpractice.
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Objective To explore the effect and safety of preoperative impact chemotherapy in the treatment of advanced gastric cancer patients.Methods 104 patients with advanced gastric cancer were randomly divided into two groups,including 54 cases of observation group,54 cases of control group.The observation group was given modified FOLFOX7 regimen for 2 courses before operation,and 6 courses FOLFOX7 regimen after operation.The control group was given routine operation and FOLFOX7 regimen for 8 coures.The effective rate,l-year,2-year,3-year survival rate and toxic effects after operation were observed.Results In observation group,the effective rate was 59.3％,the curative resection rate was 81.5％,and the overall resectability rate was 90.7％,and those was 35.2％,59.3％,75.9％ in control group,all the difference was statistically significant( x2 =8.55,6.39,4.27,all P ＜ 0.05 ).The 1 -year,2-year survival rates after operation were not significantly different ( x2 =0.38,2.06,all P ＞0.05 ),while the difference was significant at 3-year after operation( x2 =4.06,P ＜ 0.05 ).There was no significant difference on the toxic effects between the two groups ( P ＞ 0.05 ).Conclusion Modified FOLFOX7 regimen is effective and well-tolerable for patients with advanced gastric cancer,and it could contribute to improve the overall resectability rate and survival rate after operation.
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Objective To improve the skills and level of TURP and peri-operative period managements to the high risky senile patients with large volume benign prostatic hyperplasia.Methods The clinical data of 318 high risky and senile patients whose ages above 80ys,ASA score ＞ 2 and prostate volume ＞ 60g were analyzed retrospectively.They underwent the treatment of TURP.ResultsTotal 318 patients underwent TURP were safe.The operating time ranged from 40 to 85 minutes,averaged 58.2minutes;the volume of blood transfusion ranged from 200ml to 600ml;No serious complications happened during and after the operation.With follow up of 1 ～ 12 months,the International Prostate Symptom Scores(I-PSS) decreased 14.7 averagely,Quality of Life(QOL) decreased 3.3 averagely,Maximal flow rate( Qmax ) increased 6.4ml/s averagely,and Post-voided Residual(PVR) decreased 85.3ml averagely.Conclusion The actions including sufficient preparations and evaluations pre-operatively,the maintenance to the stability of circulatory system during the operation,guaranteeing the demands of blood exchange in the vital organs such as heart,lungs and brain,are the key points to the success.
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ObjectiveTo improve the treatment of urethral stricture and evaluate the applicative effects of wire-guided supporting catheterization in clinical urology. MethodsDuring January 2004 and December 2005, the patients with urethral stricture were dwelled with the catheters using the traditional guideless catheterization (group A). If the dwelling failed,it will be replaced by the wire-guided supporting catheterization using the improved catheters. During January 2006 and December 2009, the patients with urethral stricture were dwelled with the improved catheters using wire-guided supporting catheterization ( group B) straightly. Compared the first-time dwelling success incidence,the incidence of catheter associated urinary tract infection and side-effect events. Then after 1 year, compared the urethral stricture recrudescence and the course of treatment. ResultsThe success incidence of first-time catheterization in group A was 48.98％ (24/49) ,and in group B was 97.94％ (95/97) ,there was significant difference between 2 groups;The incidence of catheter associated urinary tract infection in group A was 12.24％, and 8. 25％ in group B, there was significant difference between 2 groups; As for the course of treatment, group A was 46. 2w,group B was 32.7w;The urethral stricture recrudescence in group A and B were 16.33％ and 9.28％ respectively,and there was significant difference between 2 groups. ConclusionThe wire-guided supporting catheterization, which minimize the injury and simplify the operation of internal urethrotomy,could makes the improved catheter easy to induct and replace, improve the success rate of first time-catheterization and prevent the false tunnel damage and new scar expansion. It could make benefit to reduce of incidence of catheter associated urinary tract infection and urethral stricture recrudescence,but also could shorten the course of treatment significantly.
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BACKGROUND: Looking for effective measures to ensure the survival of the implanted stem cells against ischemia-induced hypoxia becomes the major concern in the research of cell transplantation therapy for cerebral infarction.OBJECTIVE: To study the effects of human Persephin gene transfer on hypoxia-induced apoptosis of neural stem cells.DESTGN: A randomized controlled basic study on cells.SETTTNG: Department of Neurology, the Second Affiliated Hospital of Sun Yat-sen University.MATERTALS: This study was completed in the Lin Baixing Laboratory Center of the Second Affiliated Hospital of Sun Yat-sen University from July to December in 2006. Recombinant adenovirus pAdCMV persephin was constructed in our lab. C17.2 neural stem cells were kindly provided by Prof. Snyder, Harvard Medical University, USA. Trypsin and DMEM/F12 were purchased from Gibco Company (USA), fetal bovine serum (FBS) from Sijiqing Biological Engineering Materials Co. Ltd (Hangzhou, China); Poly-lysine from Sigma Company (USA), TUNEL assay kit and FuGENE kit from Roche Molecular Biochemicals Company (Swiss), and S-P immunohistochemical detection kit and DAB reaction kit from Mycine Biological Engineering Company (Fujian). Rat anti-human monoclonal Nestin antibody and rabbit anti-human polyclonal persephin antibody were manufactured by Santa Cruz Company (USA), and persephin anti-senseoligodeoxynucleotide (ODN) was synthesized by Shanghai Biological Engineering Company.METHODS: ① Interventions: C17.2 neural stem cells cultured in vitro were infected by recombinant adenovirus containing persephin gene, and they were divided into four groups: blank control group (Group A, in which the C17.2 neural stem cells were not treated with hypoxia), hypoxic group [Group B, in which the cells were cultured at 37 ℃ in anaerobic incubation containing N2 (0.95 in volume fraction) and CO2 (0.05 in volume fraction)], hypoxia + pAdCMV persephin infection group [Group C, where the cells were cultured under the conditions as in group B after pAdCMV persephin infection for 48 hours], and hypoxia + pAdCMV persephin infection + anti-sense persephin ODN group (Group D, where the cells were infected by pAdCMV persephin and anti-sense persephin ODN. ② Evaluation: The expression of Persephin protein was analyzed using Western blotting; Apoptotic index was detected with terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) assay; The changes of apoptotic rate was determined with flow cytometry.MAIN OUTCOME MEASURES: Expression of Persephin protein; Apoptotic index; Apoptotic rate.RESULTS: ① Expression of Persephin protein: A specific band (relative molecular mass of 24 000) was detected by Western blotting in pAdCMV persephin infected cells, suggesting the successful expression of persephin gene.Interestingly, the cells infected with both pAdCMV persephin and anti-sense persephin ODN also showed the specific band of about 24 000, but with much less density, indicating that anti-sense persephin ODN could effectively inhibit the expression of pAdCMV persephin. However, this band was not presented in the blank control groups. ② Apoptotic index:The apoptotic index in group C was significantly lower than those in groups B and D (P＜0.01), but still higher than that of group A (P＜0.01), suggesting that persephin gene transfer could attenuate apoptosis to some extent. ③ Apoptotic rate: The apoptotic rate in groups B and D were obviously higher than that in group A (P ＜ 0.01), and it was lower in group C than in groups B and D (P＜0.01).CONCLUSION:Recombinant adenovirus can efficiently mediate Persephin gene transfer into C17.2 neural stem cells,resulting in high expression of the exogenous Persephin in vitro, which effectively reduces C17.2 neural stem cell apoptosis induced by hypoxia.
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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.
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BACKGROUND: At present, the researches on murine model of acute radiation encephalopathy are still in investigation, and mature model making method is not clear.OBJECTIVE: To establish a murine model of acute radiation encephalopathy in order to provide a good foundation for further researches of radiation encephalopathy(REP) mechanism and therapy.DESIGN: Randomized controlled study based on the experimental animals.SETTING: Animal laboratory in a university hospital. The Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: The experiment was completed in the Laboratory of the Second Affiliated Hospital of Sun Yat-sen University from June 2001 to August 2002. Totally 30 female and 30 male SD rats, weighted(300 ± 30) g, were selected from the Animal Experiment Center of Sun Yat-sen University, and randomly divided into blank control group with 20 rats and experimental group with 40 rats.METHODS: Fourty rats' brain received 60Co γ-ray irradiation with the dosage of 7 Gy/time per day for 6 consecutive days with the total dosage of 42 Gy. The amount of ingestion and drinking, general activities, central nervous system(CNS) symptoms and signs were recorded every day. The hairs and skin of irradiated field and weight were checked and recorded weekly. On the 3rd, 7th, 14th, 30th days after radiation, the brain tissue was collected and the histopathologic changes were examined.Histopathologic changes after radiation.RESULTS: Since the third day, the ingestion and drinking amount of irradiated rats were decreased. The general activities were increased for the first two days, but decreased without abnormal nervous signs on the 3rd day. The rats in experimental group had a slower weight gain than those of control group, and the difference between them was of no statistical significance. All rats had slight alopecia and neuronal necrosis 2 weeks after irradiation.CONCLUSION: The irradiation method is reliable, practical and good for modeling REP process, which can be used in preventing or reducing the harm effect of radiation therapy on brain tissue.
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ObjectiveTo investigate the therapeutic effects of insulin o n acute cerebral infarction.MethodsEighty-six non-diabetes mel litus patients with acute middle cerebral artery (MCA) infarction were divided i nto two groups in random fashion:42 cases (control group) were treated by routin e methods and 44 cases (insulin group) by regular insulin on the basis of routin e methods.12～16U regular insulin per diem was administered by intravenous dripp n g in 7 days.Plasma glucose,insulin sensitivity indice (ISI) and score values of the European Stroke Scale (ESS) were observed.ResultsThere wer e no statistically significant differences between the 2 groups at baseline.The everyday plasma glucose levels of the insulin group.were within the normal range but were significantly lower than those of the control group (P
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AIM: To explore the structural changes of brain microvasculature and mechanism in microvascular lesion after focal cerebral ischemia with reperfusion. METHODS: Using the techniques of immunohistochemical staining, in situ hybridization,optical microscopy and transmission electron microscopy, the expression of uPA, uPA mRNA, and changes in miocrovascular structure were examined in ischemic focus and perifocal areas after focal cerebral ischemia 2 hours with various time points of reperfusion in stroke-prone renovascular hypertensive rats (RHRSP). RESULTS: The brain edema and hemorrhage were severe 12 hours to 3 days after reperfusion. Ultrastructural change showed that the damage characterizations of the basement membrane were degradation, defection, and exfoliated of basement membrane, while uPA, which attack the basememt membrane around cerebral capillaries and extra-cellular matrix, and uPA mRNA expression increased significantly in ischemic and perifocal areas 12 hour to 3 day after reperfusion. CONCLUSION: The main pathologic mechanism of brain edema and hemorrhage after cerebral ischemia with reperfusion may result from the basement membrane lesion of brain microvasculature. The increase in the expression of uPA in reperfusion area may be the main cause of the basement membrane lesion .
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AIM: To investigate the relation between the changes of the cerebral microvasculature and the expression of plasminogen activator inhibitor-1 (PAI-1) in ischemic focal and perifocal areas after 2 hours focal cerebral ischemia with reperfusion in rats. METHODS: The changes of cerebral microvascular structure were observed by optical microscope and electric microscope, the expression of PAI-1 were assessed by immunohistochemical staining, Western blotting in ischemic focus and perifocal areas after focal cerebral ischemia with reperfusion. RESULTS: The edema of extra-cellular matrix and the hemorrhage of extravessels in ischemic focus and perifocal areas were most severe, and degradation and the defect of basement membrane were also observed after 6 hours and 3 days reperfusion following focal cerebral ischemia. The expression of PAI-1 decreased significantly compared with control group (P