Objective To observe spinal cord neurotoxicity injury induced by mixture of bupiv-acaine-lidocaine compared with mixture of ropivacaine-lidocaine.Methods Forty-eight SD rats,weig-hing 250-300 g,were randomly allocated into 6 groups (n =8):NS group (group N),1.065% bupi-vacaine group (group B),1.5% ropivacaine group (group R),5% lidocaine group (group L),mix-ture group of 5% lidocaine and 1.065% bupivacaine (group LB)or mixture group of 5% lidocaine and 1.5% ropivacaine (group LR).The rats were given intrathecal cathetering after anesthesia.Tail-flick test (TFL)and the mechanical withdrawal threshold (MWT)of rats in each group were meas-ured 1 d before operation (the baseline values)and 1 d,2 d,3 d and 4 d after intrathecal administra-tion.The movement of rat double hind legs were observed at 10 min,30 min,1 h,2 h,3 h and 4 h and 1 d,2 d,3 d and 4 d after intrathecal administration,and the changes of recovery time of lower limbs were recorded.The rats were sacrificed 4 days postoperatively followed by observation on the indexes mentioned above,and spinal cord and cauda equine were taken for staining.Then the patho-logical changes of spinal cord were observed under light and electron microscope,and caudal nerve was rated for pathological injury.Results TFLs of group LR were obviously longer than those of group N,L,B,R,LB 1-4 d after intrathecal administration (P <0.05);Compared with the group N,there was no statistical difference of TFL in the group L,B,R,LB;there was no statistically sig-nificant difference of rat MWT in comparison among the six groups at each time point.There was no statistically significant difference among six groups in recovery time of movement after intrathecal ad-ministration;and the pathological injury scores were obviously higher in group LR than that of group N,L,B,R,LB (P <0.05),respectively.Compared with the group N,there was no statistical difference of the pathological injury scores in the group L,B,R and LB,respectively.Under light microscopy observa-tion:the structure of spinal cord was normal in group N,B and R.In group L and LB,there was focal ede-ma of neurofibers.In group LR,there was edema of neurofibers or demyelination,accompanied with white mass edema in posterior horn of spinal cord.Under electron microscope,only slight edema of medullated fi-bers and unmyelinated nerve fiber was seen in group B and group R;part of the medullated fibers appeared sparse focal lamellar structure in group L and group LB;swelling and degeneration of neuraxon,separation of neuraxon from myelin sheath and disappearance of unmyelinated nerve fiber were seen in group LR. Conclusion The mixture of 5% lidocaine with 1.5% ropivacaine is more neurotoxic to spinal cord than the single administration of lidocaine or ropivacaine,moreover it is not obviously changed compared with the mixture of lidocaine with equipotent bupivacaine.

Objective To observe the expression of DR5 in the ocular tissues of uveitis rats induced by endotoxin,and study the relationship between the apoptosis of inflammatory cells and expression of TRAIL / DR5.Methods SD rats were randomly divided into 3 groups:Blank control group,normal saline injection group and endotoxin injection group.The endotoxin injection group was injected with lipopolysaccharide into the rat posterior foot pad to make endotoxin-induced uveitis animal model.There were no operations in the blank control group,and the subgroups were divided into 6 hours,12 hours,24 hours and 48 hours groups according to the time of injection.The ultrastructural changes of inflammatory cells and endothelial cells in iris capillaries were observed by transmission electron microscopy (TEM).The expression of DtR5 protein on inflammatory cells at different time after endotoxin induction was detected by SABC method.Results TEM showed that the microvilli of the capillary endothelial cells in the iris tissue of the blank control group and saline injection group had more obvious vesicles with no obvious abnormal structure and shape.The number of swallowed vesicles in the capillary endothelial cells injected with endotoxin was decreased at 6 hours group,and the number of vesicles in the infiltrating neutrophils and lymphocytes decreased.Neutrophils and lymphocytes appear chromatin condensation,vacuolar changes in the expression of apoptosis.Immunohistochemistry showed that the DR5 protein was negative in the iridocular epithelium of the blank control group and saline injection group.In the endotoxin injection group,the DR5 protein was weakly colored in the iris pigment epithelium and appeared on the inflammatory cells.The number of staining and the intensity of coloring in the 24 hours group were significantly higher than those in the 6 hours group,and the color density was 0.085 9 ± 0.019 6,there were statistical differences compared with 6 hours group,12 hours group and 48 hours group (all P ＜ 0.05).Conclusion TRAIL and its receptor DR5 may be involved in the apoptosis of inflammatory cells in endotoxin-induced uveitis.

BACKGROUND: Family factor is the main reason for children's psychological and behavioral problems. Seen from the view of systemic family theory, in any circumstances, children's behavioral problems reflect various interpersonal systemic networks among society, school, family, relatives and friends and the relationship of complicated interactions among individuals.Family therapy is an important way to cure children's behavioral problems.OBJECTIVE: To observe whether systemic family rehabilitation is adequate for intervening in children's behavioral problems.DESIGN: Cluster sampling; Contrast observationSETTING: Department of Psychiatry, Kunming Medical College.PARTICIPANTS: A total of 276 students in grade four from Mingtong primary school of Kunming City were selected along with their parents from May 2004 to March 2005. A survey was conducted on students and their parents according to the principle of cluster sampling.METHODS: Self-rating family dynamics scale was used to investigate the characteristics of family dynamics of 276 students in grade four. Achenbach child behavior checklist for parents was adopted to investigate the situation of their children' s behaviors. 57 cases of children with behavioral problems were detected out, and took the families in which the whole family members voluntarily participated in the intervention as interventional group and the families in which any one of the family members did not want to participate in the intervention as abnormal control group. The students and their parents in interventional group were treated with 4-month systemic therapy. 4 months later, resent the above rating scales.MAIN OUTCOME MEASURES: Comparison of family dynamics and children's behaviors between interventional group and abnormal control group before and after intervention.RESULTS: After intervention, changes of family dynamics and children's behaviors in interventional group had taken place, which had statistical difference. The changes as follows: family atmosphere changed from "depressive and hostile" to "harmonious, pleasant, equal and exoteric"; the degree of individuation of family members changed from low level to high; disease concept changed from "regarding the patients as helpless victims" to "regarding them as helpful doers", which realize "softening of disease concept". The total scores of Achenbach child behavior checklist for parents decreased [The scores of questionnaire for fathers before and after intervention were (42.20±22.58) and (28.95±21.90) respectively, and scores of questionnaire for mothers before and after intervention were (50.95±14.60) and (36.40-±15.36) respectively, P ＜ 0.05]. There was no significant difference between the changes of children's behaviors assessed by fathers and mothers in abnormal control group [The scores of questionnaire for fathers before and after intervention were (38.32±17.30) and (37.57±16.45) respectively, and scores of questionnaire for mothers before and after intervention were (45.54±16.02) and (40.14±17.95) respectively, P ＞ 0.05].CONCLUSION: Using the idea and technique of systemic family therapy ,it is feasible and effective to intervene in children's behavioral problems by family therapy.

Objective To explore the differentiation of rat bone marrow mesenchymal stem cells into dopaminergic neuron.Methods The mesenchymal stem cells were isolated from bone marrow. They were induced to differentiate into dopaminergic neurons by freated with bFGF, VitC and EGF at the third generation. Dopamine-associated protein and genes in the treated cells were examined by immumofluorescence and RT-PCR. Dopamine in the supernatant and endoplasm from culture system was determined by ELISA kit. Results The results showed that tyrosine hydroxylase, dopamine transporter and nerve neucleoprotein and Nestin,Nurr-1 genes were found. And the dopamine existed in the supernatant and cytoplasm from inducing culture system. Conclusion The rat bone marrow mesenchymal stem cells have the capacity of differenting into dopaminergic nurons.

Phenylketonuria (PKU) is a severe autosomal recessive disease which can cause irreversible damage to patients' neural system and results in severe mental retardation.Although the institution of a lowphenylalanine (Phe) diet has been a remarkable success in preventing the devastating damage associated with untreated PKU,there are always small but consistent gap in intelligence quotient (IQ) scores and executive functioning when compared to siblings or healthy age-related control groups.During the past few years,several types of new treatment strategies,such as genetic engineering,enzyme replacement,tetrahydrobiopterin (BH4),large neutral amino acids (LNAA),low-Phe diet and liver or liver cell transplantation therapies,have been studied and improved.This paper aims to introduce the research advances in pathogenesis of PKU,the treatment methods and the related molecular mechanism.

Objective To investigate the status of medication adherence of secondary prevention after acute ischemic stroke and influence on prognosis in Qingdao area , including antithrombotic drugs , lipid-lowering drugs , antihypertensive drugs and glucose-lowering drugs , to provide the basis for making medical policy.Methods We examined patients with acute cerebral infarction and transient ischemic attack in the Department of Neurology of Affiliated Hospital of Qingdao University from December 2014 to January 2016.Patients′medication status and recurrence of stroke events were registered by using telephone and clinic follow-up within six months after the patients discharged from hospital .The standard of good and bad drug adherence was as follows:good adherence was defined as proportion of days covered ( PDC) ≥80%, bad adherence was defined as PDC <80%.SPSS 19.0 statistical software was used to analyze the influence factors of medication adherence and the influence of medication adherence on prognosis .Results Finally, 444 cases (88.62%) were analyzed.A total of 352 cases (79.28%) had high medication adherence at six months after discharging from hospital .The following factors can improve the adherence of drug treatment:history of diabetes (108 cases (30.68%) in good medication adherence group , 16 cases (17.39%) in poor medication adherence group,χ2 =6.401, P=0.011), having employee health insurance (186 cases (52.84%) in good medication adherence group , 33 cases (35.87%) in poor medication adherence group ,χ2 =8.405, P=0.004), endovascular stent implantation (29 cases(8.24%) in good medication adherence group, 0 case in poor medication adherence group ,χ2 =8.109, P=0.004), staying in hospital more than 10 days ( 230 cases ( 65.34%) in good medication adherence group , 49 cases ( 53.26%) in poor medication adherence group ,χ2 =4.558, P=0.033).Six months later , the modified Rankin Scale ( mRS) score of poor medication adherence group was significantly higher than that in good adherence group ( mRS score≥3,50 cases (14.20%) in good medication adherence group , 22 cases (23.91%) in poor medication adherence group,χ2 =5.060, P=0.024) .After six months, a total of 13 cases had recurrent cerebral infarction, with two cases ( 0.57%) in good adherence group , 11 cases ( 11.96%) in poor adherence group.High medication adherence was an independent protective factor of recurrent stroke ( OR=0.042, 95%CI 0.008 -0.210, P<0.01 ) .At one, three, six months after discharging from hospital , the medication adherence of antihypertensive and glucose-lowering drugs was better than that of antiplatelet agents and lipid-lowering drugs (all P<0.05).Conclusions The persistence and adherence to secondary preventive medication in ischemic stroke patients was generally well at 6th month after discharging from hospital.History of diabetes , having employee health insurance , stent implantation and longer hospital stay are the influencing factors to high medication adherence .High medication adherence is an independent protective factor for ischemic stroke recurrence .The medication adherence of antihypertensive and glucose-lowering drugs is better than that of antithrombotic drugs and lipid-lowering drugs.

OBJECTIVE: To investigate the self-renewal mechanism of CD133+ cancer stem cells from Hep-2 cell line. METHOD: The CD133+ cells were sorted by flow cytometry from Hep-2 cell line. Then the sorted CD133+ cells were cultured in RPMI1640. The ability of self-renewal of CD133+ cells were tested by MTT assay. mRNA and protein expression of self-renewal related genes were detected by western blot and RT- PCR. RESULT: (3.10 ± 0.21)% of Hep-2 cells expressed the membrane antigen CD133. CD133+ fraction was raised to (90.20 ± 5.51)% by flow cytometry. In vitro culture and growth curve showed CD133+ cells had more active proliferation ability than CD133- cells, which showed statistically significant difference between these two group (P < 0.01). RT- PCR and western blot results showed upregulated mRNA and protein expression of Fas, c-myc, survivin in CD133+ group (P < 0.01). In the same time, the ratio of Bcl-2/Bax gene expression was obviously increased in CD133+ group. Self-renewal related gene such as β-catenin, SHH, SMOH and Bmi-1,Gli-1 were all up-regulated in CD133+ group both in mRNA and protein. On the contrary, PTCH gene was down-regulated. CONCLUSION CD133 positive cells are a small proportion of a Hep-2 cell line. The results of this experiment verified that CD133 positive cells owned the properties of cancer stem cells. Upregulated anti-apoptotic gene is the foundatiom of self-renewal mechanism of CD133+ cells. Cancer stem cells related signal pathways such as Hedgehog, Wnt and Bmi-1 pathway are in state of activation. The identification of self-renewal mechanism about cancer stem cell provides a powerful tool to investigate the tumorigenic process in the larynx and to develop therapies targeting to these signal pathways.
AC133 Antigen ; Antigens, CD ; Apoptosis ; Cell Physiological Phenomena ; physiology ; Down-Regulation ; Flow Cytometry ; Glycoproteins ; Humans ; Laryngeal Neoplasms ; Neoplastic Stem Cells ; physiology ; Patched Receptors ; Patched-1 Receptor ; Peptides ; Receptors, Cell Surface ; genetics ; metabolism ; Signal Transduction ; beta Catenin ; genetics

BACKGROUND:Zinc can improve bone reaction to titanium implants through regulation of osteoblast activity under the condition of osteoporosis. OBJECTIVE:To study the zinc ions effect on improving the stability of titanium implants under the condition of osteoporosis. METHODS:After removal of the ovary, 28 female Sprague-Dawley rats presented with osteoporosis symptoms for 12 weeks and then, they were randomly divided into two groups. Two kinds of titanium implants with (experimental group) or without zinc (control group) were inserted into the bilateral femoral metaphysis longitudinal y in the ovariectomized rats, separately. Three rats were selected from each group for double fluorescence labeling examination. Mineral apposition rate was calculated at postoperatively 6 weeks. Another four rats were selected from each group for histomorphometry analysis at postoperatively 12 weeks. Biomechanical test was in the remaining rats in the two groups to calculate the maximum push-out force and maximum shear force at 12 weeks postoperatively. RESULTS AND CONCLUSION:The double fluorescence labeling examination revealed that the mineral apposition rate was improved in the experimental group as compared with the control group (P<0.05). New bone formation occurred in the two groups. However, larger new bone amount, higher bone area ratio and bone-to-implant contact were detected in the experimental group (P<0.05 or P<0.01). The maximum push-out force and maximum shear force were also superior in the experimental group to the control group (P<0.05 or P<0.01). These findings indicate that zinc ions are capable of promoting bone formation and improving implant fixation in ovariectomized rats.

BACKGROUND:The rapidly developed transplantation of peripheral blood stem cells have been successfully used to substitute bone marrow transplantation and become the first choice method for transplantation of hemopoietic stem cells.It is relatively difficult to collect peripheral blood stem cells from young age and low body mass infants.OBJECTIVE:To investigate the safety and adverse effects of peripheral blood stem cell collection from young age and low body mass through the use of blood cell separator.METHODS:Two type 1 diabetes mellitus infants,aged younger than 2 years old and with body mass less than 15 kg,were treated using autologous hemopoietic stem cell transplantation.The two infants were adequately comforted to lesion the fear of collection.At 1 week prior to collection,calcium agent was orally taken to reduce the incidences of low calcium.Within 24 hours prior to collection,oily food was forbidden,and on the day of collection,milk was forbidden,to avoid chylemia,which influences blood collection.Prior to collection,200 mL 25 Gy y-ray radiated red blood cells suspension was injected into the tube,which was routinely placed in the subclavian vein,to avoid hypovolaemic syndrome and the effects on hematocrit.Individualized collection parameters were set,During collection,blood circulation volume was 3,4 times of systemic blood circulation to ensure sufficient total circulation volume.During isolation,the ratio of ACD to whole blood was kept between 1:11 and 1:13 to prevent sodium citrate poisoning.RESULTS AND CONCLUSION:Peripheral blood stem cells were successfully collected during first intention in each infant.During collection,stable vital signs but no adverse effects were observed.After collection,mononuclear cells weighted 14.71×108/kg and 18.82×108/kg respectively,and CD34+ cells were about 34.13×108/kg and 32.38×106/kg,respectively in each infant.Therefore,it is feasible to collect peripheral blood stem cells from infants of young age and low body mass under sufficient psychological preparation.

Objective To observe the effects of pulsed electromagnetic fields (PEMFs) on behavior and mechanical paw withdrawal thresholds (MPWTs) after acute skeletal muscle contusion (ASMC) in rats, and to in-vestigate the application of PEMFs in rats with ASMC during the early stage. Methods Forty-two Sprague-Dawley rats were randomly divided into a PEMF group (P group) , control group (C group) and blank control group ( BC group). ASMC models were set up in groups P and C, and no intervention was applied in the BC group. A PEMF was administered to animals in the P group immediately after the ASMC was inflicted. The behavior of the rats in each group was then observed. The MPWT of each rat was tested 2 days before and 0, 12, and 18 hours after the ASMC was inflicted). Results In the P and C groups, MPWT of the left hind paw at the 12th and 18th hour after ASMC was significantly lower than the baseline pain threshold 2 days before the ASMC. At 18 hours, the MPWT was signifi-cantly higher than at 12 hours in the P group. MPWT at 12 hours in the P group and at both 12 and 18 hours in the C group were significantly lower than in the BC group. MPWT in the P group at 18 hours was significantly higher than in the C group. Conclusions The behavior of rats treated with PEMF immediately after ASMC was improved, and their pain threshold was still elevated 18 hours later.