1.Experimental study on the transplantation of neural stem cells derived from bone marrow of crab-eating macaque
Yiquan KE ; Gang LI ; Ruxiang XU ; Xiaodan JIANG ; Wenping CHENG
Chinese Journal of Tissue Engineering Research 2005;9(5):200-201
BACKGROUND: Researches indicated that bone marrow stem cells (BMSCs) could differentiated into neural stem cells in vitro, but what was the role of neural stem cells(NSCs) in the recovery of cortical injury,whether the NSC is capable of growing and migration in injured still remained unknown.OBJECTIVE: To explore the growing state of autograft NSC derived from crab-eating macaque BMSC transplanted in brain.DESIGN: Prospective case control study based on experimental animals.SETTING: Department of neurosurgery in a hospital of a military medical university.MATERIALS: This study was carried out at Center Laboratory of Neurological Research Institute, Zhujiang Hospital affiliated to the First Military Medical University of Chinese PLA. Six healthy adult crab-eating macaques were purchased from the South China Primate Animal Center.INTERVENTIONS: BMSCs harvested from six crab-eating macaques were cultured in vitro and induced to differentiate into neural stem cells, which then labeled by bromodeoxyuridine(BrdU) and autografted into brains.MAIN OUTCOME MEASURES: Brain tissues underwent hematoxylin and eosin(HE) staining and immunohistochemical staining before observed under optical microscope.RESULTS: The results of HE staining showed that the cell number in injured brain vas obviously higher in both instant and delayed transplanting groups than sham-transplanting group; moreover cells were proved reacting to BrdU by immunohistochemical staining in cortical injuries of both groups at 1-6 months following stem cells autograft, as well as at neighboring white matters at half year later, but no BrdU positive cells could be found in traumatic controls, sham-transplanting group and normal brains.CONCLUSION: NSCs derived from in vitro cultured BMSCs were proved capable of surviving, proliferating, differentiating and migrating in cortex after autograft, so that BMSCs is considered as replacing cells or the source of NSCs; moreover autograft stem cells could survive, proliferate and migrate in old cortical traumatic focus.
2.Relationship of somatosensory evoked potential grading with prognosis in long-term unconscious patients after severe traumatic brain injury
Xiangyu WANG ; Weiwei XU ; Yueling ZHOU ; Xiaodan JIANG ; Yiquan KE
Chinese Journal of Trauma 2010;26(4):314-317
Objective To study prognosis and grading of somatosensory evoked potential(SEP)in long-term unconscious patients after severe traumatic brain injury(TBI).Methods Five prognostic factors including age,sex,injury mechanism,history of temporal craniotomy and SEP grading were selected and analyzed in 47 patients after severe TBI with a duration of unconsciousness longer than two weeks.The prognosis was judged by Glasgow Outcome Scale.Results Prognosis was closely associated with SEP grading(P=0.024).The accuracy of SEP in assessing the prognosis was 91.5%.About 95%-100% of patients with SEP at grade Ⅲ-Ⅲ ended up with severe disability,persistent vegetative state or death.However,43.75% of patients with SEP at grade Ⅰ had good prognoses.Conclusions The SEP grading can objectively and accurately evaluate patients' prognosis and demonstrate the brain function.
3.Comparison of HC visual laryngoscopy and fiberoptic bronchoscope guided endotracheal intubation in patients undergoing cervical surgery
Hongfei CHEN ; Yiquan WU ; Yujian ZHANG ; Kejian SHI ; Xuzhong XU
Journal of Chinese Physician 2016;(z1):26-29
Objective To compare the clinical effects of HC video laryngoscope and fiberoptic bronchoscope (FOB)in guidance of endotracheal intubation for patients undergoing cervical surgery.Meth-ods A total of 50 patients (ASA I or II)with cervical vertebra injury,nerve root cervical spondylopathy or cervical spondylotic myelopathy in the first affiliated hospital of Wenzhou medical university were selected, all of whom were undergone selective cervical operations between March 2014 and June 2015.The patients were randomly divided into two groups (n =25):HC video laryngoscope group (group H)and FOB group (group F).After induction of anesthesia,HC video laryngoscope and FOB were used for tracheal intuba-tion.⑴ Intubation time and success rates of intubation were recorded,and visual analogue scale (VAS) was used to evaluate the difficulty of intubation;⑵ Mean arterial pressure (MAP),heart rate (HR),and rate-pressure product (RPP)before incubation (Tb),immediate after intubation (T0),and 1min after in-cubation (T1)were recorded;(3)Intubation-related complications were recorded.Results ⑴ The dura-tion of intubation (19.7 ±7.1 )s in the group H was significantly shorter than that of group F (51.9 ± 19.2)s (P <0.05).The one-time success rate of intubation in group H (92%)was higher than that of group F (64%),with statistically significant differences (P <0.05).Intubation was easier in group H (P <0.05).⑵ MAP,HR and RPP of group H at T0 were higher than those of group F.And at T1,there were no statistical differences between groups in MAP,HR and RPP (P >0.05).⑶ The incidence of complications (sore throat)was lower in group H (P <0.05).Conclusions Compared to FOB,HC vid-eo laryngoscope-guided selective cervical operation has the characteristics of higher success rate of intubat-ion,shorter operation time,and easier operation,more stable hemodynamics and lower incidence of intuba-tion complications.Therefore,HC video laryngoscope is a safe and effective method in the intubation for pa-tients undergoing cervical surgery.
4.Cerebral hippocampal neuronal apoptosis following kainic acid-induced epilepsy and the intervention of antagonists of dopamine D1 and D2 receptors
Songqing WANG ; Haitang CHEN ; Yiquan KE ; Ruxiang XU ; Xiaodan JIANG ; Yiran ZHANG ; Lifeng CHEN
Chinese Journal of Tissue Engineering Research 2005;9(25):250-253
BACKGROUND: Dopamine is closely associated with occurrence of epilepsy and transmission in central nerval system, and its various functions are determined by specific receptors.OBJECTIVE: To establish temporal epilepsy model so as to probe into the influences of SCH23390, the antagonist of dopamine D1 receptors and haloperidol, the antagonist of dopamine D2 receptors injected in substantia nigra on temporal epileptic seizure induced by kainic acid and on electroencephalic activityDESIGN: Randomized controlled verified experiment.SETTING: Neurology Medicine Institute of Zhujiang Hospital Affiliated to Southern Medical University.MATERIALS: The experiment was performed in General Military Neurology Medicine Institute of Zhujiang Hospital Affiliated to First Military University of Chinese PLA from August to December 2004, in which, 30SD adult male rats were employed, massed varied from 250 to 300 g.METHODS: ① 30 rats were randomized into physiological saline (control) group (6 rats), kainic acid (KA) group (6 rats) and experimental group (18 rats). The experimental group was divided into 3 subgroups, named the antagonist of dopamine D1 receptors, SCH23390 + kainic acid group (D1 +KA group), the antagonist of dopamine D2 receptors,haloperidol + kainic acid group (D2+KA group) and physiological saline + kainic acid group (PS + KA group), 6 rats in each. In the control, physi ological saline 2 μL was injected in the right cerebral ventricle unilaterally. In KA group, kainic acid 2 μL was injected in the right ventricle. In each of experimental group, SCH23390, the antagonist of dopamine D1 re ceptors, haloperidol, the antagonist of dopamine D2 receptors and physio logical saline 1 μL for each was injected in substantia nigra on the right side successively and simultaneously, kainic acid 2 μL was injected in the right ventricle. ② Observed items: alters of EEG on the 0.5th 1st, 2nd, 6th and 24th hours after medication in each experimental group (compared with EEG of non-epileptic behavior, appearance of sharp wave, spike wave,sharp (spike) slow comprehensive wave and multi-spike slow wave determines epileptic activity) and changes in animal behaviors (0 grade: normal; Ⅰ grade: wet dog-like trembling, paroxysmal facial spasm, like winking,beard moving, rhythmic chawing; Ⅱ grade: rhythmic nodding; Ⅲ grade:paroxysmal spasm of anterior limbs; Ⅳ grade: paroxysmal spasm of bilateral anterior limbs when standing; Ⅴ grade: falling down, loss of balance and convulsion of four limbs). Cerebral hippocampal neural cell apoptosis was observed and the rats were sacrificed on the 5' day of medication. Cerebral hippocampal section was prepared and determined after in situ end labeling staining.MAIN OUTCOME MEAUSRES: ① Changes in behavior in rats before and after epilepsy and electroencephalogram (EEG) alters. ② Results of cerebra hippocampal neural cell apoptosis.RESULTS: Thirty rats entered result analysis. ① Epilepsy seizure: In the control group, there was no epilepsy attacked. In KA group, all of rats ap pear seizure, which attacked 10 minutes after KA injected in brain ventricle, reached the peak in 1 hour and stopped in 3 to 6 hours. ② EEG record: In the control group, there was not epileptic activity manifestations,like sharp wave, spike wave, spike slow comprehensive wave, etc. In KA group, epileptic wave presented in 10 minutes after injection, the seizure developed to the peak in about 1 hour, the wave amplitude was decreased in 3 to 6 hours, presenting paroxysmal slow and spike slow waves and no epileptic wave appeared after 12 hours. ③ Neuronal apoptosis: In the control group, few neural cell apoptosis was visible in hippocampus after injection.In KA group, neural cell apoptosis was visible obviously in hippocampus in 5 days after injection (P =0.00). With SCH23390, the antagonist of dopamine D1 receptors, hippocampal cell apoptosis was not reduced remarkably (P >0.05) and with haloperidol, the antagonist of dopamine D2 receptors injected in substantia nigra, hippocampal cell apoptosis was aggravated (P =0.00).CONCLUSION: Injection of SCH23390, the antagonist of dopamine D1 receptors in substantia nigra cannot block kainic acid inducing epilepsy and epileptic electroencephalic activity is not weakened remarkably. Injection of haloperidol,the antagonist of dopamine D2 receptors enhances epileptic electroencephalic activity in kainic acid induced epilepsy and increases cell apoptosis remarkably in cerebral hippocampal CA3 area.It is to explain that it is dopamine D2 acceptor that is involved in regulation of temporal epilepsy in substantia nigra rather than D1 acceptor.
5.Differentiative capability of bone marrow mesenchymal cells into neural stem cells in cats
Yiquan KE ; Wenping CHENG ; Gang LI ; Xiaodan JIANG ; Ruxiang XU ; Songqing WANG
Chinese Journal of Tissue Engineering Research 2005;9(22):233-235
BACKGROUND: Bone marrow mesenchymal cells, multiple-potential non-hematopoiefic stem cells adhering to the wall in vitro culture, can be induced to proliferate and differentiate towards neurons and glia cells.OBJECTIVE: To investigate the growth state of cat bone marrow mesenchymal cells in vitro culture, as well as the capability to differentiate towards neural stem cells.DESIGN: A randomized sampling study.SETTING: Department of Neurosurgery, Zhujiang Hospital, Southern Medical University.MATERIALS: This study was carried out at the Central Laboratory of the General Military Neurological Research Institute, Zhujiang Hospital, Southern Medical University between January and December 2002. Twenty healthy home-raised cats, aged 1.0 - 2.0 years and weighing 2. 5 - 4.0 kg, male and female in half, were provided by the Animal Center of the First Military Medical University of Chinese PLA.INTERVENTIONS: Bone marrows were randomly aspirated from the left or right hindlimbs in order to separate bone marrow mesenchymal cells, then the bone marrow mesenchymal cells single cell suspension was co-cultured with neural stem cell culture media in vitro so as to induce differentiation to neural stem cells with tretinoin. CK2 type inverted optical microscope(Olympus,Japan) was used to observe the growth of bone marrow mesenchymal cells in vitro culture, as well as 4, 12, 24, 48 hours of induction upon eliminating or not eliminating the wall-adhering cells. Bone marrow mesenchymal cells in stem cell stage were identified under Olympus optical microscope with modified immunohistochemical staining.MAIN OUTCOME MEASURES: The growth state and the immunocytochemical staining of living bone marrow mesenchymal cells exposed to experimental intervention were observed under the Olympus inverted optical microscope.RESULTS: Data from the 20 cats were analyzed without loss. Reversed microscopic observation revealed that cat bone marrow mesenchymal cells becrame larger when cultured in vitro, which were rich in plasmic granules with prominence projecting, adhering to the wall and forming cell clones. These cells were then successively cultured, and imnunohistochemical staining analysis suggested that the passaged bone marrow mesenchymal cells could express neural stem cells-specific antigen Nestin and differentiate towards glia-like cells and neuron-like cells.CONCLUSION: Cat bone marrow mesenchymal cells possess the characteristics of stem cells; they can be amplified into cell clones and induced to express the property of neural glia cells and neuron-like cells under proper condition.
6.Study on the molecular mechanism of the dissemination of a novel CTX-M-like extended spectrum beta-lactamase-producing Escherichia coil
Jiehua JIANG ; Weijiao LIAO ; Jianyun YI ; Tao CHEN ; Xiuxin SUN ; Yunjian XU ; Yiquan LI
Journal of Chinese Physician 2008;10(7):882-885
Objective To investigate the distribution of the CTX-M- extended spectrum beta-lactamase (ESBLs) producing Esche- richia coli(ECO) and the molecular mechanism of dissemination. Methods To analyze the drug resistance of the 43 isolates, Kirby-Bauer susceptibility method was used. Multiple polymeraso chain reaction (PCR) was used to amplify the gene of ESBLs, AmpC, full length of blaCTX-M-like gene, insertion sequence (IS) ISEcp1B, IS903 , IS26 and integron I. NEST-PCR was used to detect if the beta-lactamase gene lo- cated in the integron I. The product of full length of bla-CTX-M like gone amplified by PCR was sequenced. Results Susceptibility test showed the resistance from high to low in turn was Ampicillin (97.68%), Coftriaxone (67.44 % ), piperacillin(65.12 % ), Cefotaxime (62.79 % ) ,Coftasidime(58.14% ), Cofasolin(55.81% ), Cofepime (53.49%), Cefexitin(51.16%), ciprofloxacin (44. 19% ), Aztreo- nam(41.86% ), Cefoperasone/Sulbactam ( 20.93% ), Amikacin (0% ), Imipenem (0% ), respectively. ECO was susceptive to Imipenem. CTX-M-G1 was found in 25 strains of ECO , TEM, SHV, CTX-M-G1, ISEcp1B, and integron I were found in the nine isolates. IS903 were found in ECO 3 and 5, and IS26 was found in ECO 3. In ECO 3 and 5, blaCTX-M-like was flanked upstream by ISEcp1B element that provided -35 and -10 promoter sequences and a right inverted repeat (IRR) recognized by transposase, downstream by IS903 provided an inverted re- peat, ISEcp1 B and IS903 composed the complex transpeson. Conclusion ISEcplB may drive the expression and dissemination of blaCTX-M-like gene at a high level.
7.Relationship between serum ferritin and nonalcoholic fatty liver diseases in obese children
Yiqing CHU ; Yanping WAN ; Renying XU ; Xiaomin ZHANG ; Yiquan ZHOU ; Zhiqi CHEN ; Liping LU
Chinese Journal of Clinical Nutrition 2015;23(5):266-270
Objective To investigate the relationship between serum ferritin and nonalcoholic fatty liver diseases in obese children.Methods Obese children aged 6 to 14 years old were enrolled.Duration of obesity, anthropometric parameters (height, body weight, waist circumference, hip circumference), bioelectrical impedance analysis (body fat), serological parameters (liver transaminases, lipid metabolism, fasting blood glucose, fasting insulin, serum ferritin) and liver ultrasonography were recorded.Insulin resistance (IR) index was calculated by homeostasis model assessment (HOMA).All subjects were divided into 3 groups according to liver ultrasound and liver transaminases : simple obese children (SOC) group, obese children with nonalcoholic simple fatty liver (NAFL) group and obese children with nonalcoholic steatohepatitis (NASH) group.Results 86 obese children entered the study, with a mean age of (10.4 ± 1.9) years, including 26 in the SOC group, 28 in the NAFL group and 32 in the NASH group.Waist circumference standard deviation score (SDS or Z-score), waist-to-hip ratio, HOMA-IR index and serum ferritin in the NASH group were obviously higher than those in the NAFL group [2.3 ± 0.3 vs.2.1 ± 0.3, P =0.020;1.0 ± 0.0 vs.0.9 ± 0.1,P=0.014;4.0±1.7 vs.2.9±1.8, P=0.006;(104.1 ±49.6) μg/Lvs.(68.4 ±22.7) μg/L, P=0.004] and the SOC group [2.3 ±0.3 vs.1.9 ±0.3, P=0.000;1.0±0.0vs.0.9 ±0.1, P=0.012;4.0 ±1.7 vs.2.5 ±1.6, P=0.001;(104.1 ±49.6) μg/Lvs.(59.2 ±28.9) μg/L, P=0.001], while there was no significant difference in body mass index Z-score [2.8 ± 0.5 vs.2.7 ± 0.6, P =0.524;2.8 ± 0.5 vs.2.7 ± 0.6, P =0.662].There were no significant differences between the NAFL group and the SOC group in the above indicators [2.1 ±0.3 vs.1.9 ±0.3, P =0.260;0.9 ±0.1 vs.0.9 ±0.1, P =0.952;2.9 ± 1.8vs.2.5±1.6, P=0.283;(68.4±22.7) μg/Lvs.(59.2±28.9) μg/L, P=0.161].Mter controlling age, body mass index, waist circumference, waist-to-hip ratio, triglyceride, and HOMA-IR index, serum ferritin was still positively correlated with the magnitude of nonalcoholic fatty liver diseases in obese children (r =0.335, P =0.002).Conclusion Serum ferritin is probably an independent risk factor for NASH in obese children.
8.Comparison of degree of pain in patients after radical gastrectomy under different anesthetic regimens
Yiquan WU ; Zhousheng JIN ; Qimin LIU ; Fangfang XIA ; Fuli LIU ; Xili DING ; Huimin DONG ; Xuzhong XU
Chinese Journal of Anesthesiology 2012;32(1):74-77
Objective To compare the degree of pain in patients after radical gastrectomy under different anesthetic regimens.Methods One hundred and two ASA Ⅰ or Ⅱ patients of both sexes,aged 50-75 yr,weighing 45-70 kg,undergoing elective radical gastrectomy,were randomly divided into 3 groups ( n =34 each):general anesthesia (GA) group,combined general-subcostal transversus abdominis plane block (CGTA) group and combined general-epidural anesthesia (CGEA) group.The patients were sent to the postanesthesia care unit (PACU) after tracheal extubation,and the VAS score on arrival in the PACU was recorded.The degree of pain was evaluated by VAS score,and when VAS scores > 3,the patients received intravenous morphine titration.When VAS scores ≤ 3,morphine titration was stopped and all the patients were connected to patient-controlled intravenous analgesia and/or epidural analgesia pump.The total amount of morphine consumed was recorded at the end of titration,and the occurrence of adverse reactions was also observed.Results Compared with groups GA and CGTA,the incidence of moderate to severe postoperative pain was significantly decreased in group CGEA (P <0.01).The incidence of severe postoperative pain,the VAS score on arrival in the PACU and the total amount of morphine consumed were decreased gradually in groups GA,CGTA and CGEA ( P < 0.01 ).The incidence of sedation was significantly lower in group CGEA than in group GA (P < 0.01 ).There were no significant differences in the other adverse reactions among the three groups ( P > 0.05 ).Conclusion The degree of pain is reduced gradually in patients after radical gastrectomy under GA,CGTA and CGEA.
9.Retrospective analysis of risk factors of colonization of central venous catheters
Yiquan ZHOU ; Renying XU ; Yanping WAN ; Xiaomin ZHANG ; Zhiqi CHEN ; Liping LU
Chinese Journal of Clinical Nutrition 2013;21(6):355-358
Objective To investigate the risk factors of central venous catheters (CVC) colonization.Methods A retrospective study was performed on adult patients with CVCs placement in Renji Hospital,Shanghai Jiaotong University from January 2006 to March 2010.Clinical data,catheter-related information (including duration of catheter placement,position and purpose of catheterization,and whether or not out-of-ward catheterization),catheter culture results,and prevalence of catheter-related blood stream infection (CRBSI)was collected.Results A total of 651 patients aged 18 to 97 years (median:63 years) were enrolled in the study,in whom 762 CVC were placed.The median duration of catheter placement was 1 1 days (2 to 122 days)and the total duration of CVC placement was 10 725 days.The prevalence of catheter colonization was 16%(122/762),and 134 germs were cultured.Gram-positive cocci was the most common colonized bactera (52.2%,70/122),followed by gram-negative bacilli (33.6%,45/122) and fungi (14.2%,19/122).Overall 13 CRBSI were confirmed and the rate of CRBSI was 1.21/1000 catheter-days.Logistic regression analysis demonstrated that the risk factors for CVC colonization included mechanical ventilation [odds ratio (OR) =1.783,95% confidence interval (Cl) =1.108 ~2.870],serum albumin concentration less than 25 g/L before catheterization (OR =1.783,95% Cl =1.357 ~ 6.757),prolonged duration of catheter placement (OR =1.105,95% Cl =1.009 ~ 1.111),and out-of-ward catheterization (OR =2.837,95% Cl =1.010 ~7.969).Conclusion Patients with prolonged duration of catheter placement and out-of-ward catheterization are inclined to CVC colonization.
10.Preparation of antisense oligodeoxynucleotides-loaded butylcyanoacrylate nanoparticles by interfacial polymerization and their stability
Yue XU ; Yiquan KE ; Xixiao YANG ; Xiqing YAN ; Jianqi WANG ; Lesong HUANG
Journal of Third Military Medical University 2003;0(14):-
Objective To optimize the preparation of nanoparticles encapsulating antisense oligodeoxynucleotides in a-butyleyanoacrylate carrier (ASODN in NP) and investigate their stability. Methods ASODN in NP were prepared by interfacial polymerization of butyleyanoacrylate (BCA). The formulation and technology of the prepared NP was optimized by using orthogonal design based on the single-factor experiment. The morphology of NP was examined by transmission electron microscope; The size and size distribution of NP were determined by Malvern laser granularity equipment;The encapsulation efficiency and drug loading were determined by HPLC; The ability of protecting oligodeoxynucleotides from serum was investigated on a 20% polyacrylamide-7 Murea sequencing gel (PAGE). Results The nanoparticles in the optimal conditions were of regular spherical surface and discrete. The average size was 97.1 nm,the average encapsulation efficiency and drug loading of ASODN in NP were 96.7% and 10.1% respectively; The oligonucleotides were more efficiently protected from degradation by nucleases than by oligonucleotides adsorbed into nanospheres.Conclusion ASODN in NP has good stability,encapsulation efficiency,drug loading and great potential for ASODN delivery.