Objective To investigate the effect of molecular chaperone on the prokaryotic expression level and activity of tetanus toxin fragment C(TTC) in order to provide new ideas for the development of new tetanus vaccines.Methods The gene sequence of TTC was optimized for E.coli adaptive codons,and the recombinant plasmid pET30a-HC was constructed,which was identified by double enzyme digestion.The recombinant plasmid pET30a-HC and molecular chaperone pGro7were co-transformed into competent E.coli BL21(DE3) and induced by IPTG to express TTC.The obtained TTC was identified by 12% SDS-PAGE and Western blot,and purified by Ni FF metal chelate affinity chromatography column.Female Kunming mice were immunized with TTC,TTC+Al,TTC+CpG,PBS,and tetanus toxoid(TT) by subcutaneous injection,10 mice in each group.After immunization,blood samples were collected by tail cutting or eyeball picking,and the serum was separated.The antibody titer,antibody typing,IL-4 and IFNγ levels were detected by ELISA.The lung and liver of mice were harvested aseptically,and the safety of TTC was evaluated by histopathological method.Results The recombinant plasmid pET30a-HC was constructed correctly as identified by double enzyme digestion.After adding molecular chaperone pGro7,the soluble expression of the target protein accounted for 40% of the total supernatant expression.After 42 days of the initial immunization,the specific IgG antibody titer in serum of TTC+CpG group was significantly higher than that of PBS,TTC,and TT groups(t=4.753-8.624,each P <0.01),which was also higher than that of TTC+Al group,but there was no significant difference(t=0.966,P> 0.05).There was no significant difference in serum IgG1,IgG2a,IgG2b,IgG3,and IgGM antibody levels among the groups(t=0.163-0.203,P> 0.05),IgG1/IgG2a> 1,indicating that the immune pathway was biased towards Th2 type.Compared with TTC group,the serum IL-4 and IFNγ levels in TTC+Al group had no significant difference(t=2.666 and 2.470,respectively,each P> 0.05),while significantly increased in TTC+CpG group(t=18.080 and8.440,P <0.001 and <0.05,respectively).Compared with TT group,the serum IL-4 level in TTC+CpG group increased significantly(t=8.561,P <0.01),but there was no significant difference in IFNγ level(t=0.472,P> 0.05).No obvious pathological changes were found in lung and liver of mice in each group.Conclusion Molecular chaperone can improve the soluble expression level of TTC with good safety,and combined with CpG adjuvant,it can induce strong immune response.

Objective To study the change of glutamate transporter (GluTs) function of hippocampus in seizures rats induced by KA, and explore the role of GluTs in the mechanism of epilepsy.Methods 60 adult male Wistar rats were randomly divided into two groups: KA group(Ⅰ group) and control group(Ⅱ group). Each group was randomly divided into five subgroups according to different time after kindling, including 4 h, 24 h, 48 h, 5 d,and 7 d after injection. GluTs function was studied by means of assay of 3H-L-glutamate uptake in synaptosomes and tissue sections of hippocampus.Results The function of GluTs in synaptosomes was singnificently decreased at any time (all P

OBJECTIVETo explore the feasibility and safety of dexmedetomidine combined with remifentanil for lower eyelid blepharoplasty.

METHODS50 patients undergoing lower eyelid blepharoplasty ASA I - II were randomly divided into 2 groups (25 in each group) as observation group and control group. Dexmedetomidine (0.3 microg x kg(-1) x h(-1)) and remifentanil (0.05 microg x kg(-1) x min(-1)) were used in observation group. Local anesthesia by lidocaine (1%) combined with midazolam (0.04 mg/kg) intravenous sedation were used in control group. HR, mean arterial pressure (MAP), heart rate (HR), breathing rate (RR), pulse oxygen saturation (SpO2) change were monitored and recorded before and after the administration of the drug, as well as at the beginning of operation, during the operation, at the end of the operation, and 5 minutes after the operation. Patients sedation degree was evaluated by Ramsay. Operation time, recovery time, the incidence of adverse reactions, the satisfaction of patients and doctors were also recorded.

RESULTSPatients had no obvious difference of intraoperative MAP, RR, SpO2, operation time and postoperative recovery time between the two groups (P > 0.05). Compared with control group, HR during the operation were decreased significantly [(64.2 +/- 8.2) bpm, P < 0.05] in observation group. In control group, there were 18 cases of patients with agitation, but they endured the operation anyway. In observation group, 24 cases of patients underwent operation without agitation. The satisfaction of patients and the operation doctors (96%) in observation group were markedly higher than that in control group (P < 0.05).

CONCLUSIONDexmedetomidine combined with remifentanil is practical and safe. The effect of reducing HR should be noticed.

Adult ; Blepharoplasty ; Dexmedetomidine ; therapeutic use ; Double-Blind Method ; Female ; Humans ; Hypnotics and Sedatives ; therapeutic use ; Male ; Middle Aged ; Piperidines ; therapeutic use

Objective To analyze results of bacterial infection distribution and drug sensitivity monitoring in 414 patients with open extremity injuries caused by Wenchuan earthquake so as to provide advices for guiding utility of antibiotics in treatment of patients injured in natural disasters. Methods Bacterial culture of wound secretion substance and drug sensitivity monitoring were done in 414 patients with open extremity injuries. Results There were 12 patients (2.9%) with G+ coccobacteria, 66 (15.9%) with G- bacillus, 2 (0.5%)with G+ bacillus, 6 (1.5%) with multiplicitas infection(5 with dual infection and I with triplication infection). Main pathogenic bacteria were as follows: (1)G+cocco- bacteria including Staphylococcus epidermidis, Staphylococcus aureus and anthropo-staphylococci ; (2) G- bacillus including Aerobacter cloacae, banmanii, escherichia co]i, aerugo pseudomonas, proteusvul- gaffs and Ewingella americana. (3)G+ bacillus including bacillus subtilis. The drugs with high sensi0tivi- ty to G+ coccobacteria included vancomycin, rifampin, levofloxacin and cidomycin, but those with low sensitivity to G+coccobacteria included penicillin, ampicillin and erythremycin. The drugs with high sen- sitivity to G- bacillus included imipenem, amikacin, levofloxaciu and tenebrimycin, but those with low sensitivity to G-bacillus included ampicillin and piperacillin. Conclusions Patients with open ex- tremity injuries caused by earthquake have rather high bacterial infection rate, involving many kinds of in- fection strains. Therefore, we should do bacterial culture and drug sensitivity monitoring of wound excre- tion substance as early as possible to guide rational use of antibiotics.

According to the widely use of peripheral inserted center catheters , this paper reflects ethical issues form the aspects such as the evaluation before catheter is lack of standards , the access and appraisal system is lax , the continuation of care does not reach and lack of humanistic care aspects .Furthermore , the article points out that the scientific assessment , grasping the indications strictly , strengthening the norms , stricting access system , taking the initiative to follow -up, strengthening health education , the professional ethics , the humanistic care and so on, can ensure the safety of patients and improve nursing service level as a whole .

Objective To explore the interleukin (IL)-6,IL-10 and T cell subsets levels in bronchoalveolar lavage fluid(BALF) of children with refractory mycoplasma pneumonia.Methods A total of 53 children with refractory mycoplasma pneumonia were selected as the observation group,30 children with bronchial foreign body in our hospital were chosen as controls during the same period.ABC-double antibody sandwich ELISA method was used to detect IL-6,IL-10 levels and the CD3 +,CD4 + and CD8 + T levels were measured by means of flow cytometry in BALF.Results The IL-6 and IL-10 levels in BALF of children in the observation group were (63.25 ± 18.61) ng/ml,(31.83 ± 8.33) ng/ml respectively,and they were significantly higher than those of the controls[(30.51 ± 1.34) ng/ml,(11.01 ± 2.91) ng/ml] (P ＜ 0.05,respectively).The percentage of CD3 +,CD4 +,CD8 + T cells and the ratio of CD4 +/CD8 + T cells in BALF of the observation group were (48.47 ± 2.88)％,(21.16 ± 6.29)％,(23.04 ± 4.63)％,0.94 ± 0.33,respectively,and they were significantly lower than those of the controls [(64.24 ± 3.06) ％,(34.34 ± 7.59) ％,(26.71 ±5.29)％,1.56-±0.67] (P＜0.05,respectively).Conclusion The IL-6,IL-10 levels in BALF of children with refractory mycoplasma pneumonia significantly increased,suggesting that cell-mediated immunity play an important role in the pathogenesis of refractory mycoplasma pneumonia.

Aim To investigate the effects of midazolam on porcine isolated coronary artery rings pre-contracted by potassium chloride(KCl)and the possible mechanism.Methods The vessel tension recorder system was used.Isotonic tension of porcine isolated coronary artery rings precontracted by KCl(30 mmol·L-1)was recorded.The vasorelaxing action of midazolam and effects of various drugs were observed in the rings.Results Midazolam(3×10-6～1×10-4 mol·L-1)respectively concentration-dependently reduced the contraction induced by KCl,and there was significant difference between the rings with intact and denude endothelium(P<0.05).On KCl-induced precontraction,midazolam′s relaxation was depressed by L-NAME and the blend of L-NAME and L-Arg(P<0.05),but was not affected by Indo,L-Arg and 1400W.The contraction was not prevented by pretreatment with the inhibitor of Na+/Ca2+ exchanger(KB-R7943).The inhibitor of KATP(Gli)restrained the diastolic function of midazolam(P<0.05),while the inhibitor of BKCa(TEA),Kir(BaCl2),KV(4-AP)had no obvious effect.Conclusions Midazolam produces remarkable vasodilatation on KCl pre-contracted porcine isolated coronary artery rings.Its relaxtion effect is via concentration-dependent and endothelium-dependent mechanisms and relevant to the production of NO.Na+/Ca2+ exchanger is not involved midazolam′s vasodilatation on KCl pre-contracted porcine coronary artery rings.The relaxant mechanism of midazolam may be concerned with KATP.The Kir,BKCa and KV may be not involved.

Objective To study the effects of extracted active components of Chaenomeles Speciosa (EACCS) on non-alcoholic fatty liver disease (NAFLD) in mice; To discuss the possible molecular mechanism. Methods Forty male KM mice were randomized into four groups, namely normal group, model group, low-dose (50 mg/kg) EACCS group and high-dose (100 mg/kg) EACCS group. Except that the normal group was daily given routine diet, the other groups were given high-fat–high-fructose diet (HFFD). The mice were put to death 4 weeks later. Body weight, liver weight and serum TG were measured. HE and oil red O staining were used to observe liver tissue morphology. RT-PCR and Western blot were used to detect the expression of lipid metabolism related genes. Results Compared with the normal group, the liver size, liver index (P<0.01) and epididymal fat index (P<0.05) increased significantly;The ALT and GLU in serum increased (P<0.05), TG increased (P<0.05), and pathological findings showed significant steatosis; RT-PCR and Western blot showed that the expression levels of SIRT1 and FoxO1 mRNA decreased and the level of SERBP-1c increased in the model group. Compared with the model group, the hepatic lipid accumulation of EACCS groups was obviously improved, and the serum ALT, GLU, and TG levels significantly decreased, the expression levels of hepatic SIRT1 and FoxO1 mRNA increased. Conclusion EACCS has protective effects on NAFLD mice induced by HFFD, and its mechanism may be related to the activation of SIRT1-FoxO1 signaling pathway in the liver tissues.

Objective: To understand physical health of pre school children aged 3-6, so as to analyze the early life influencing factors, and to provide a reference for physical health promotion of preschool children. Methods: The method of random sampling was used to select 1 580 preschoolers aged from 3-6 in Guangzhou. National physical health test and questionnaire survey were conducted to assess physical health. Results: The physical fitness pass rate of preschool children was 87.2%, among which the excellent rate was 30.8%. With the increase of age, the rate of good physique of both boys and girls increased. There were statistical differences in body shape indexes (except dermal fold thickness) between male and female children at all ages ( P <0.01). The rate of overweight and obesity was 14.7%, which was higher in boys (17.2%) than in girls (12.0%). There were significant differences between overweight and non overweight obese people( χ 2=4.84，3.96，5.73，4.85， P <0.05). Logistic regression showed that preterm birth was negatively correlated with the rate of good physique of preschool children ( OR=0.81, P <0.05). There was a positive correlation between birth weight greater than gestational age and the rate of physical fitness of preschool children ( OR=1.37, P <0.05). Conclusion Preschool children aged 3-6 years old in Guangzhou have poor physical health overall, and the rate of good physical health is related to factors such as preterm gestational week in early life delivery and birth weight greater than the gestational age.

Objective To investigate the clinical significance of piatelet activity in patients with chronic cor Dulmonale.Methods PAC-1 and CD62p was measured with flow cytometry in whole blood samples from 40 patients and 30 normal controls.The pulmonary arterial pressure was detected through Doppler echocardiography.The arterial partial pressure of oxygen and the carbon dioxide were also analyzed.Results PAC-1 and CD62p increased significantiy (P<0.01).Conclusion Platelet activity is positively related to pulmonary artrial systolic pressure,CO2 partial pressure,and negatively related to O2 partial pressure.