BACKGROUND: The neural stem cells (NSCs) will be clinically applied extensively in the future, and seeking of more promoting methods of the proliferation and differentiation of NSCs in vitro will be the study direction of NSCs.OBJECTIVE: To introduce an economic and time-saving method for NSCs proliferation.DESIGN: Observation and control experiment.SETTING: Beijing Neurosurgical Institute.MATERIALS: Four Wistar rats pregnant for 14 days with the body mass of (180±20) g (bought from Animal Department of Chinese Academyof Medical Sciences with the batch number of SCXK1100-0006 for experiment animals) were selected and fed at common temperature and humidity.The DMEM/F12 and B27 were bought from Gibco Corporation. The basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) were bought from PeproTech Company. The nidogen monoclonal antibody (MA),glial fibrillary acidic protein (GFAP) polyclonal antibody, galactocerebroside polyclonal antibody and microtubule-associated protein 2 (MAP2) were obtained from Chemicon Company. The fetal bovine serum (FCS) was provided by Hyclone company.METHODS: The experiment was conducted in the Department of Injury and Repair of Beijing Neurosurgical Institute from May to October 2004.Wistar rats were executed by dislocation and sterilized by putting into 75% alcohol. Four rats were used each time, the brain tissues of which were put into Hank's fluid. The cerebral pia mater and vessels were isolated under anatomical microscope, and the brain tissues were sheared off with eye scissors, which were filtered by 200 mesh copper grid and collected into 2 centrifuge tubes, and the supernatant was gotten rid off after that. ①Cells were divided into serum pre-culture group and control group according to whether there were serum pre-culture. The DMEM nutrient fluid of FCS (100 g/L) was added to serum pre-culture group, which was replaced by DMEM/F12 nutrient fluid of EGF, bFGF and B27 at 48 hours after culture. The cells in the control group were added with DMEM/F12 nutrient fluid of EGF, bFGF and B27 to culture in 5% CO2 incubator for one week at 37 ℃. The growth of NSCs at 48 hours after culture was observed in both groups under inverted contrast phase microscope.②On the 5th and 10th day after differentiation induced by 100 g/L FCS, nestin, GFAP, galactocerebroside and MAP2 were stained with immunofluorescence antibody,and the expressions of NSCs in both groups were observed under fluorescent microscope. The PBS buffer solution was used instead of first antibody in the control group, while other procedures were the same as above.MAIN OUTCOME MEASURES: ①Observation of NSCs growth at 48 hours after culture under contrast phase microscope in both groups. ②Detection of NSC specific antigen expressions with immunofluorescence stain in both groups.RESULTS: ①In the cells of the serum pre-culture group at 48 hours after culture, there were large number of regular NSC bulbs, most of which were aggregated by 8-10 cells. The proliferation of cell bulbs was accelerated after the nutrient fluid was replaced to DMEM/F12 nutrient fluid of EGF,bFGF and B27, while irregular lamellar cells could be seen in the control group at 48 hours after culture, and small regular cell bulbs were found at 4-5 days later. ②It could be seen under fluorescence microscope that on the 5th day after induced differentiation, the nestin, GFAP and galactocerebroside were positive, while MAP2 was negative. On the 10th day after induced differentiation, nestin and GFAP were positive, whereas the galactocerebroside and MAP2 were negative (no representation).CONCLUSION: Serum pre-culture can accelerate the bulb-aggregation of NSCs as well as promote the proliferation of NSCs.

Objective To observe the influence of health education on compliance of children nn-dergoing megacolon radical operation. Methods A questionnaire investigation was conducted on compli-ance of children undergoing megncolon radical operation before and after health education. ResultsCompliance of children patients and their family members increased after health education (P<0.05).Conclusions Health education could improve the compliance of children patients and their family mem-bers as well as the cognition about the disease knowledge so that the rehabilitation was facilitated.

Objective To determine the optimum dose of pentazocine when combined with propofol for gastroscopy in elderly patients.Methods One hundred and forty ASA physical status Ⅰ or Ⅱ patients,aged 6575 yr,scheduled for elective gastroscopy under general anesthesia,were randomly assigned into Ⅰ-Ⅳ groups (n =35 each) using a random number table.Before insertion of the gastroscope,pentazocine 0.2,0.4 and 0.8 mg/kg were injected intravenously in Ⅱ-Ⅳ groups,respectively,while the equal volume of normal saline was given in Ⅰ group.Propofol was then administered by target-controlled infusion (TCI).The half-effective concentration (EC50 of propofol was determined by up-and-down sequential trial.The target plasma concentration (Cp) was set at 3.5 μg/ml in the first patient.Gastroscopy was performed at 5 min after the target effect-site and plasma concentrations were balanced.The response to gastroscopy was defined as positive when body movement and/or bucking occurred during gastroscopy.Each time the Cp increased/decreased by 0.3 μg/ml in the next patient depending on whether or not the response to gastroscopy was positive.EC50 and 95 ％ confidence interval of propofol TCI inhibiting the response to gastroscopy were calculated using Probit analysis.The development of respiratory depression and hypotension was observed.Results EC50 (95 ％ confidence interval) of propofol TCI inhibiting the response to gastroscopy was 2.82 (2.63-3.02) μg/ml,2.78 (2.58-2.97) μg/ml,2.16 (2.00-2.32) μg/ml and 2.03 (1.88-2.19) μg/ml in Ⅰ-Ⅳ groups,respectively.Compared with group Ⅰ,EC50 was significantly decreased in Ⅲ and Ⅳ groups,and the incidence of respiratory depression was increased in Ⅳ group (P ＜ 0.05).Conclusion The optimum dose of pentazocine when combined with propofol is 0.4 mg/kg for gastroscopy in elderly patients.

This study is to establish the gastric hot model of rats. After gastric feeding with ethanol solution for 3 weeks and feeding with extra capsaicin and ethanol solution for another 2 weeks, model group show distinct physical sign of gastric hot syndrome. The pathology of gastrics reveals gastricism of model group, while treatment group (treat with Zuojin Wan) shows mild lesion. Elisa detection of model group show that the solution of interleukin-2 (IL-2) is higher than the blank group. The obvious difference among model group, treatment group and blank group reveals the success of the establishment of gastric hot model.
Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Eating ; Female ; Humans ; Male ; Rats ; Rats, Wistar ; Stomach Diseases ; drug therapy ; pathology ; physiopathology

Objective To evaluate the effects of dexmedetomidine on lung ischemia-reperfusion (I/R) injury in rats.Methods Ninety-six healthy SPF male Wistar rats,weighing 250-350 g,aged 8-12 weeks,were randomly divided into 4 groups (n =24 each):sham operation group (S group),I/R group,low dose dexmedetomidine group (DL group) and high dose dexmedetomidine group (DH group).In DL and DH groups,dexmedetomidine 100 and 500 μg· kg-1 · d-1 were injected intraperitoneally once a day for 2 consecutive days,while the equal volume of normal saline was given in S and I/R groups.Lung·I/R was induced by clamping the left hilum of lung for 45 min followed by reperfusion at 30 min after administration on 2nd day.At 45 min of ischemia,and 60 and 120 min of reperfusion,6 rats were sacrificed,and lungswere removed for determination of TNF-α (tumor necrosis factor-a) content and myeloperoxidase (MPO) activity in lung tissues.The percentage of damaged alveolar in lung tissues was detected at 120 min of reperfusion.Another 6 rats were lavaged and the broncho-alveolar lavage fluid (BALF) was colleted for determination of the total protein concentrations.Results Compared with S group,the TNF-α content,MPO activity,percentage of damaged alveolar,and total protein concentrations in BALF were significantly increased in I/R,DL and DH groups.Compared with I/R group,the TNF-α content,MPO activity,percentage of damaged alveolar,and total protein concentrations in BALF were significantly decreased in DL and DH groups.Conclusion Dexmedetomidine can alleviate the lung I/R injury in rats,and the mechanism is related to inhibiton of the inflammatory responses.

BACKGROUND:Finding of neural stem cells(NSCs)brings new hope for repairing central nervous system(CNS)injury.However,the influence of internal environment after brain injury on the survival and differentiation of NSCs is a complexand variable process.OBJECTIVE:To observe the survival and differentiation of human embryonic NSCs following implantation into rats with fluid percussion brain injury.DESIGN:Open experiment.SETTING:Department of Neurosurgery,Guangdong Provincial Hospital of Traditional Chinese Medicine;Beijing Institute of Neurosurgery.MATERIALS:This experiment was carried out In the Laboratory of Neural Stem Cells,Beijing Institute of Neurosurgery from September 2002 to March 2003.Twenty-four female SD rats,aged 7 weeks,with body mass of(250±10)g,were provided by the Experimental Animal lnstitute, Chinese Academy of Medical Sciences[License Nd. SCXK(Jing)2002-2003]. Cerebrum of 8-week aborted fetus was obtained (Informed consents were obtained from parturients and their relatives). Fetal survival was monitored by B ultrasonic wave during abortion. BrdU monoclonal antibody(Sigma Company),rabbit anti-nidogen polyclonal antibody(Chemicon Company),mouse anti-microtubule-associated protein 2 (MAP-2)monoclonal antibody(Neomarkers Company),rabbit anti-gliaI fibrillary acidic protein(GFAP)polyclonal antibody (Biogenex Company).METHODS:① CerebraI cortex cells of 8-week aborted human fetus was harvested and cultured in vitro for obtaining human embryronic NSCs.②Rat models of hydraulic impact injury were developed.Bone window of motor sensory area of cerebral cortex was set at 2.5mm posterior to bregma which was zero point and 3.0 mm lateral to midline.Hydraulic impact injury parameters were set as impact pressure 0.3 MPa. impact time 25 ms and impact time once. ③BrdU-labeled human embryronic NSCs were implanted into injured area al 24 hours after injury.After 1 and 4 weeks.rats were sacrificed.Adjacent sections were doubly stained bv BrdU/MAP-2 and BrdU/GFAP.MAIN OUTCOME MEASURES:①Survival and immigration of implanted human embryonic NSCs.②Differentiation of implanted human embryonic NSCs.RESULTS:①BrdU-positive cells were oval and brown.At 1 and 4 weeks after implantation,BrdU-positive cells survived and migrated,and they migrated more widely at 4 weeks after implantation. ②At 1 week after implantation,more BrdU-positive cells were found in the subcortical granular layer and subcortex,and BrdU/MAP-2-positive cells were more than BrdU/GFAP-positive cells;At 4 weeks after implantation,BrdU-positive cells were significantly reduced,and found in choroid plexus and blood capillary,and BrdU/GFAP-positive cells were more than BrdU/MAP-2-positive cells.CONCLUSION:Implanted human embryonic NSCs can survive in the region of brain injury.gradually differentiate into astrocytes during rehabilitation and are easily digested by endothelial phagocytes.It indicates that immunological rejaction possibly influences the survival of human embryonic NSCs.

This study is to establish the gastric cold model of rats. After gastric feeding with cold water for 5 weeks and extra iced water bath in the last 2 weeks, model group show distinct physical sign of gastric cold syndrome. The pathology of gastrics reveals gastricism of model group, while treatment group(treated with Fanzuojin Wan) show mild lesion. Elisa detection of model group show that the solution of interleukin-2 (IL-2) is higher than blank group. The difference with significance among model group, treatment group and blank group reveals the success of the establishment of gastric cold syndrome.
Animals ; Cold Temperature ; Disease Models, Animal ; Female ; Humans ; Male ; Rats ; Rats, Wistar ; Stomach ; chemistry ; metabolism ; pathology ; physiopathology ; Stomach Diseases ; metabolism ; pathology ; physiopathology

Objective To evaluate the anti-tumour effect of 60℃ LipiodoI in the embolization of VX_2 hepatocarcinoma in rabbits.Methods VX_2 carcinoma cells were surgically implanted into the left liver lobe in 30 male New Zealand white rabbits,which were randomly divided into 3 groups by figure and table method with 10 rabbits in each group.Physiological saline,Lipiodol(37℃),and Lipiodol(60℃)were injected in each group via hepatic artery and liver cancer was embolized.The volume of tumour and serum level of aspartate aminotransferase(AST)were observed after one week,and the survival period of VX_2 rabbits was also observed.Results In the group of Lipiodol(60℃),the growth rate of tumour(0.92? 0.21)was significantly lower than that of control group(3.48?1.17)and Lipiodol(37℃)groups (1.69?0.26),respectively(F=34.95,P0.05),but was significantly higher than the control group(68.6?6.6)U/L(t=19.24,P

Objective:To evaluate the efficacy and safety of docetaxel+ prednisone in the treatment of castrated resistant prostate cancer in patients over 75 years old.Methods:In this study, 118 metastatic castration resistant prostate cancer (mCRPC) patients over 60 years old treated in Beijing Hospital from February 2013 to December 2019 were retrospectively analyzed. The median age of the patients was 72 (65, 77)years, ECOG scores ≤2. All 118 cases had bone metastasis, 5 cases had visceral metastasis. A total of 40 patients chose docetaxel as the first-line treatment of mCRPC, and the remaining 78 patients chose docetaxel as second-line or third-line treatment. The study included 53 patients >75 years old and 65 patients aged 60-75 years. The age of patients in the two groups were 67 (63, 71) years old and 78 (76, 83) years old, the difference was statistically significant ( P<0.05). Among them, there were 24 cases with Gleason score ≤7 and 41 cases with Gleason score >7 in 60-75 years old group, and 30 cases with Gleason score ≤7 and 23 cases with Gleason score >7 in the group of >75 years old, with significant difference between the two groups ( P = 0.034). Sixty-one patients received endocrine therapy and 4 received orchiectomy in the 60-75 years old group; 43 patients received endocrine therapy and 10 received orchiectomy in the group of >75 years old, the difference was statistically significant ( P=0.035). There were 37 cases with ECOG 0 score, 25 cases with 1 scores and 3 cases with 2 scores in the group of 60-75 years old; there were 5 cases with ECOG 0 score, 38 cases with 1 score and 10 cases with 2 score in the group of >75 years old, with significant difference between the two groups ( P<0.05). There was no significant difference in PSA level[ 90 (35.5, 258) ng/ml vs. 115 (60, 296) ng/ml], G8 scale score [(14.3±2.1 vs. 13.6±1.1)], Mini-Cog score[3(2, 3) vs. 3(1, 3)], and visceral metastasis [2 cases (3.1%) vs. 3 cases (5.7%)]( P>0.05). The efficacy and safety of docetaxel in the two groups were further observed. Results:The median follow-up time was 21.5 (6, 62) months. There was no significant difference in chemotherapy cycle [(6.1±1.3) vs. (6.8±1.7)] and chemotherapy dose [(70.3±4.3) mg/m 2 vs. (66.3±5.2) mg/m 2] between the 60-75 years old group and the >75 year old group ( P> 0.05). The PSA response rate [72.3%(47/65)vs.66.0%(35/53)], pain relief rate [45.0% (9/20) vs. 54.5% (6/11)], and median progression-free survival[6.1 (1.4, 11.2) months vs. 5.9 (2.0, 12.0) months] had no statistical significance ( P>0.05). There were no deaths in the two groups during chemotherapy. The median overall survival(OS) of patients aged 60-75 years and those >75 years old who received docetaxel as first-line treatment were 26.5 (16.1, 31.3) months and 24.8 (17.5, 28.4) months, respectively ( P=0.223). The median OS of the two groups were 17.3 (13.2, 20.5) months and 15.4 (12.3, 20.0) months with docetaxel treatment as second or third line treatment ( P=0.331). There were 3 cases (4.6%) and 5 cases (9.4%) of grade 3 adverse reactions in 60-75 years group and >75 years old group, respectively. Grade 3 leukopenia occurred in 1 case time (1.5%) and 2 cases (3.8%) respectively. Grade 3 neutropenia fever occurred in 1 case time in both groups. There was no significant difference in the incidence of above complications between the two groups ( P > 0.05). Conclusions:The efficacy and safety of docetaxel + prednisone chemotherapy for mCRPC patients >75 years old were similar to those of 60-75 years old. Age should not be the absolute contraindication of docetaxel for prostate cancer chemotherapy.

ObjectiveTo observe the changes of artery diameters, cerebral blood flow and glutamate level in the early stage (2 h after the stimulus finished) of acute mechanical middle cerebral artery (MCA) vasospasm in cats. MethodsThe right MCA was persistently mechanically stimulated using a small smooth stainless steel nail in the field across the olfactory tract for 30 min. The diameter of MCA was recorded with metrical ocular of microscope. The changes of the perfusion index of brain tissue were observed through the Laser Doppler flowmetry monitor fixed on the skull. The level of glutamate were investigated through high performance liquid chromatography. ResultsThe diameter of MCA decreased to 68.8% of normal. 2 h later, the diameter of MCA recovered. The perfusion index of the cortex surface decreased to 42.6% of normal and up to 63.8% 2 h later. The level of glutamate raised about 40 times of normal and maintained a high level 2 h after the mechanical stimulus. ConclusionThe persistent mechanical stimulus can cause acute cerebral vasospasm. Reduce of cerebral blood flow and raise of excitatory amino acids were observed in the early stage of acute mechanical vasospasm.