Objective To discuss the impact of the neurotoxity of bupivacaine and bupivacaine-induced cellular neurotoxicity caused by pretreatment of ganglion (GM-1 )monoglyceride on the ex-pression of caspase-3.Methods The mouse neuroblastoma cells-N2a cells was used as a research object to carry out the following experiments:(1)To observe the damage effects of different concen-trations of bupivacaine on N2a cells and find out the most suitable damage concentration to establish cell damage model.The N2a cells were interacted with bupivacaine with different concentrations [0μmol/L (group C),600 μmol/L (group B1),900 μmol/L (group B2),1 200 μmol/L (group B3), 1 500 μmol/L (group B4),2 000 μmol/L (group B5)]for 6,12,24,36 h and then evaluated by CCK-8 cell survival.Each experiment was repeated three times.The protective function of GM-1 to bupiva-caine-induced N2a cells damage.The N2a cells were treated with different concentrations (0.1μmol/L (group BG1),1.0 μmol/L (group BG2),10 μmol/L (group BG3))of GM-1 pretreatment 24 h,CCK-8 was evaluated in cell viability,Western Blot method was used to detect damaged cells caspase-3 expression levels.Each experiment was repeated three times.Results (1)Bupivacaine could significantly damage N2a cells,the greater the bupivacaine concentration,the longer the action time,the stronger neurotoxicity.(2)GM-1 bupivacaine nerve injury had a significant protective effect in a dose-related manner.The maximum of protective dose of this experiment was 10 μmol/L.Conclusion Bupivacaine can significantly damage N2a cells,correlating with both dose and time double positively,while GM-1 pretreatment significantly reduced the expression of caspase-3 induced by bupivacaine.

Objective To explore the characteristics of surface electromyogram (sEMG) of the forearm muscles of Parkinson's disease patients with or without load. Methods 26 Parkinson's disease patients and 28 healthy individuals participated in this study. All the subjects performed isometric contraction for elbow flexion without load or with a load of 1.5 kg. The sEMG signals were collected by surface elec-trodes and processed by linear time and frequency domain method. Results The values of median frequency (MF) and mean power frequen-cy (MPF) were higher in the patients than in the healthy controls without load (P<0.05). The value of average EMG (AEMG) was lower in the patients than in the healthy controls with a load of 1.5 kg (P<0.001). For the patients, the values of MF and MPF were higher without load than with a load of 1.5 kg (P<0.001), and the value of AEMG was lower without load than with a load of 1.5 kg (P<0.001). For the healthy controls, only the value of AEMG was lower without load than with a load of 1.5 kg (P<0.001). Conclusion The values of MF and MPF are higher in Parkinson's disease patients than in the healthy controls without load while the value of AEMG is lower in Parkinson's dis-ease patients than in the healthy controls with load.

Objective To investigate the therapeutic effects of intravenous monosialo ganglio-sides(GM-1)on neurotoxicity of intrathecally administered bupivacaine in rats and its possible mecha-nism.Methods One hundred and eight adult male Sprague-Dawley rats,weighing 280-300 g,were randomly divided into 3 groups (n=36 each):sham operation group (group sham),group saline and group GM-1.Neurotoxicity model was performed by injecting 0.12μl/g body weight of bupivacaine at concentrations of 5% via an implanted intrathecal catheter at 90-minute intervals for 4.5 h in groups saline and GM-1.After observing 24 h,group GM-1 was administered GM-1 30 mg/kg by intrave-nous injection for 7 days,once a day;while groups saline and sham received equal volume of normal saline.The recovery of the locomotor function was evaluated with Basso,Beattie and Bresnahan (BBB)and tail-flick latency(TFL)before injection bupivacaine and days 1,3,5,7,14,28 after in-jection,TFL was converted to the percent maximum possible effect (%MPE).Six rats were sacri-ficed in each group at each time point,and spinal cord was taken to examine histological injury scores by light and electron microscopy at the L3 level,and neuron caspase-3 expression was evluated using immunohistochemistry and RT-PCR.Results Compared with group saline,%MPE,histological inju-ry score and caspase-3 mRNA expression were decreased on days 7,14 and 28;Caspase-3 protein ex-pression was decreased on days 5,7,14 and 28;while BBB score was higher on days 14 and 28 in group GM-1 (P < 0.05 ).Compared with group sham,% MPE,histological injury score,caspase-3 mRNA and protein expression in groups GM-1 and saline were significantly higher,while BBB score was lower on 1,3,5,7,14 and 28 d after injection (P <0.05).Conclusion GM-1 can promote neuro-functional recovery after bupivacaine neurotoxicity in rats through the possible mechanism of down-regulating neuron caspase-3 expression.

Objective To evaluate the efficacy of intrathecally administered monosialoganglioside (GM-1)for treatment of bupivacaine spinal anesthesia-induced neurotoxicity to rat spinal cord.Methods Adult male Sprague-Dawley rats,weighing 280-300 g,in which the intrathecal catheter was successfully inserted into the L3,4 intervertebral space and advanced toward the tail,were randomly divided into 4 groups (n =36 each):sham operation group (group S),GM-1 group,bupivacaine group (group B) and bupivacaine + GM-1 group (group BG).In B and BG groups,the rats received 5％ bupivacaine 20 μl via the intrathecal catheter 3 times at 1.5-hour intervals.GM-1 20 μg was injected intrathecally 24 h later once a day for 7 days in BG and GM-1 groups.Before bupivacaine injection and on days 1,3,5,7,14 and 28 after bupivacaine injection (T0-T6),tail flick latency (TFL) was measured,MPE (percentage of maximal possible effect) was calculated,and the locomotor recovery was evaluated using the Basso,Beattie,Bresnahan (BBB) Locomotor Rating Scale.Then six rats were randomly chosen and sacrificed in each group.Spinal cord was removed for histopathologic examination (with light and electronic microscope) and for determination of caspase-3 protein and mRNA expression (by immuno-histochemistry and RTPCR).The pathological changes of the spinal cord were scored.Results Compared with S and GM-1 groups,MPE,pathological scores,and caspase-3 protein and mRNA expression were significantly increased and BBB score was decreased at T1-6 in group B (P ＜ 0.05),and MPE was increased at T1-5 (P ＜ 0.05) and returned to the baseline value at T6 (P ＞ 0.05) and pathological scores,and caspase-3 protein and mRNA expression were significantly increased and BBB score was decreased at T1-6 in group BG (P ＜ 0.05).There were no significant differences in each parameter at each time point between S and GM-1 groups (P ＞ 0.05).Compared with group B,MPE and caspase-3 protein and mRNA expression were significantly decreased at T2-6,pathological scores were decreased at T3-6,and BBB score was increased at T4-6 in group BG (P ＜ 0.05).The pathological changes of spinal cord tissues were obvious at T1-6 in group B and at T1-3 in group BG,but the changes gradually recovered at T4-6 in group BG.Conclusion Intrathecally administered GM-1 has therapeutic effect against bupivacaine spinal anesthesia-induced neurotoxicity to rat spinal cord,and inhibition of neuronal apoptosis in the spinal cord may be involved in the underlying mechanism.

OBJECTIVE:To establish a method for the simultaneous determination of syringinand and pedunculoside in Zhuang medicine Yuyang powder. METHODS:HPLC was performed on the column of Agilent ODS with mobile phase of acetonitrile-water (gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 210 nm,the column temperature was 30℃,and the in-jection volume was 5μl. RESULTS:The linear range was 0.71-3.55μg for syringin(r=0.999 7)and 1.62-8.10μg for pedunculoside (r=0.999 8), respectively;RSDs of precision, stability and reproducibility tests were lower than 3%;recoveries were 100.8%-104.9%(RSD=1.7%,n=6) and 96.0%-100.8%(RSD=2.2%,n=6). CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of syringinand and pedunculoside in Zhuang medicine Yuyang powder.

BACKGROUND:Stem cel transplantation has achieved good results in the treatment of cerebral ischemia, and how to reduce apoptosis of transplanted cel s has become the focus of the therapy.
OBJECTIVE:To investigate the injured effect of tumor necrosis factor alpha (TNF-α) on bone marrow mesenchymal stem cel s and its mechanism.
METHODS:Primary cultured bone marrow mesenchymal stem cel s from Sprague-Dawley rats were treated with 200μg/L TNF-αfor 6 hours. Cel vitality was assayed by MTT, and cel apoptosis was observed by Hoechst33342 staining. Apoptotic rate was detected by Annexin-V/PI double staining. Level of oxidative stress was evaluated by determination of malondialdehyde and superoxide dismutase levels. The protein expressions of phosphorylated-Akt, Akt, phosphorylated-FoxO1, FoxO1 were detected by western blot analysis.
RESULTS AND CONCLUSION:After treatment with TNF-α, the cel vitality of bone marrow mesenchymal stem cel s decreased, the apoptotic rate increased, and the cel s were arrested in the S phase. Moreover, the oxidative stress level was elevated, and the protein expression of phosphorylated-Akt and phosphorylated-FoxO1 was significantly reduced compared with the control group (P<0.05). These results suggest that TNF-αat high level contributes to the S-stage arrest, responsible for the apoptosis processes of bone marrow mesenchymal stem cel s via the Akt-FoxO1 pathway.

Objective The incidence rate of pressure ulcer is high in critical patients and off-loading mattresses and reposi-tioning are known as effective interventions for the prevention of pressure ulcers .However, evidence is lacking for selection of the right type of mattresses and suitable interval of repositioning .This study was to compare the effects of two types of off-loading mattresses with two different repositioning intervals in preventing pressure ulcers in critical patients . Methods According to the design of this ran-domized controlled trial , we made a training plan concerning the participants , methods of intervention and comparison , criteria and methods of observation , and methods of recording , and trained 26 nurses from 7 hospitals .Using non-inferiority design and the method of stratified blocked randomization , we divided 1194 patients with the risk of pressure ulcer into a trial group ( n=596) and a control group ( n=598) , a viscoelastic sponge mattress with every-four-hours repositioning used for the former and an automatic aeration mat-tress with every-two-hours repositioning for the latter , both for 7 successive days .We examined the patients every day , recorded the in-cidence and stages of pressure ulcer , and compared the data obtained between the two groups of patients . Results The total inci-dence rate of pressure ulcer was 1.09%(13/1194), significantly lower in the trial than in the control group (0.34%[2/596] vs 1.84%[11/598], P=0.012). Conclusion A viscoelastic sponge mattress with every-four-hours repositioning is superior to an automatic aeration mattress with every-two-hours repositioning and therefore is preferred to the latter in preventing the incidence of pressure ulcer in critical patients in the ICU .

Objective To evaluate the effect of ganglioside ( GM-1) preconditioning on endoplas-mic reticulum stress ( ERS)-dependent apoptosis during spinal cord injury induced by bupivacaine in rats. Methods One hundred and eight clean-grade healthy male Sprague-Dawley rats, aged 8 weeks, weighing 250-300 g, were divided into 3 groups ( n=36 each) using a random number table method: control group (group C), bupivacaine group (group B) and GM-1 pretreatment group (group G). In group B, 5％bupivacaine 0. 12 μl/g was intrathecally injected at 1. 5 h intervals, 3 times intotal. In group G, GM-120μl ( 30 mg/kg) was intrathecally injected, and 24 h later bupivacaine was intrathecally injected according to the method previously described in group B. Immediately after intrathecal injection and at 1, 3, 5, 7 and 14 days after intrathecal injection, the maximum percentage of anti-nociceptive effects (％MPE) was detected, the hindlimb motor function score ( BBB score) was evaluated, and spinal cord tissues were har-vested for determination of cell apoptosis ( using TUNEL) and expression of caspase-12 and CCAAT/enhan-cer-binding protein homologous protein ( CHOP ) protein and mRNA ( by Western blot or quantitative real-time polymerase chain reaction) . The apoptosis rate was calculated. Results Compared with group C, the％MPE and apoptosis rate were significantly increased, the BBB score was decreased, and the expression of CHOP and caspase-12 protein and mRNA was up-regulated in group B ( P＜0. 05) . Compared with group B, the ％MPE and apoptosis rate were significantly decreased, the BBB score was increased, and the ex-pression of CHOP and caspase-12 protein and mRNA was down-regulated in group G ( P＜0. 05) . Conclu-sion GM-1 preconditioning reduces bupivacaine-induced spinal cord injury possibly through inhibiting ERS-dependent apoptosis in rats.

OBJECTIVETo explore the correlation between cytogenetic findings and clinical manifestations of Turner syndrome.

METHODS607 cases of cytogenetically diagnosed Turner syndrome, including those with a major manifestation of Turner syndrome, were analyzed with conventional G-banding. Correlation between the karyotypes and clinical features were analyzed.

RESULTSAmong the 607 cases, there were 154 cases with monosomy X (25.37%). Mosaicism monosomy X was found in 240 patients (39.54%), which included 194 (80.83%) with a low proportion of 45,X (3 ≤ the number of 45, X ≤5, while the normal cells ≥ 30). Structural X chromosome abnormalities were found in 173 patients (28.50%). A supernumerary marker chromosome was found in 40 cases (6.59%). Most patients with typical manifestations of Turner syndrome were under 11 years of age and whose karyotypes were mainly 45,X. The karyotype of patients between 11 and 18 years old was mainly 45,X, 46,X,i(X)(q10) and mos45,X/46,X,i(X)(q10), which all had primary amenorrhea in addition to the typical clinical manifestations. The karyotype of patients over 18 years of age were mainly mosaicism with a low proportion of 45,X, whom all had primary infertility. 53 patients had a history of pregnancy, which included 48 with non-structural abnormalities of X chromosome and 5 with abnormal structure of X chromosome.

CONCLUSIONGenerally, the higher proportion of cells with an abnormal karyotype, the more severe were the clinical symptoms and the earlier clinical recognition. Karyotyping analysis can provide guidance for the early diagnosis of Turner syndrome, especially those with a low proportion of 45,X.

Abortion, Spontaneous ; genetics ; Adolescent ; Adult ; Amenorrhea ; genetics ; Child ; Child, Preschool ; Chromosomes, Human, X ; genetics ; Cytogenetic Analysis ; methods ; Female ; Humans ; Infant ; Infant, Newborn ; Karyotyping ; Middle Aged ; Mosaicism ; Pregnancy ; Sex Chromosome Aberrations ; Turner Syndrome ; genetics ; pathology ; Young Adult

ObjectiveTo examine the clinical features of fractures and related risk factors in patients with rheumatoid arthritis(RA) in China.MethodsSix hundred and eighty-one RA patients were randomly selected from department of rheumatology of 18 hospitals of China.Data were obtained from the questionnaire,including age,sex,disease duration,the involvement of joints,treatment regimen,features of fractures etc.The possible risk factors of fracture in patients with RA were analyzed with a multi-variate Logistic regression analysis.Results① In 681 RA patients of the survey,48 patients had 54 fractures,and the incidence of fractures was about 8％.② Fractures occurred at various sites.Foot/ankle,femur,spine and wrist were the mostfrequent sites.③ The Logistic regression analysis showed that several factors increased the risk of fracture in RA patients,including long disease duration (OR:1.245,95％CI:0.987-1.570,P=0.065),male gender(OR:0.433,95％CI:0.199-0.942,P=0.035),more deformed joints(OR:1.042,95％CI:1.006-1.079,P=0.023),family history of RA (OR:2.201,95％CI:0.984-4.923,P=0.055),and high scores of SF-36(OR:1.017,95％CI:1.002-1.033,P=0.028).④ According to the degree of correlation from strong to weak,the risk factors of fracture were disease duration,SF-36,sex,number of deformed joints and family history of rheumatoid arthritis.ConclusionThe incidence of fracture is high in patients with rheumatoid arthritis.Several factors could increase the risk of fractures in RA patients,including long disease duration,male gender,more deformed joints,and family history of RA.In order to prevent the occurrence of fractures,cautions should be taken to prevent the development of fractures and treat the disease aggressively to suppress the disease activity of RA.