Objective:To observe the effects of preemptive analgesia with parecoxib sodium on differentiation of peripheral T helper cells(Th) in patients with radical mastectomy .Methods :A toal of 50 patients with radical mastectomy were randomly di-vided into parecoxib group(Group P) and the control group(Group C) .Parecoxib sodium(40 mg ,diluted with normal saline into 5 mL ,the same below ) was intravenously injected 1 h before operation in Group P ,and 12 h after the first injection .The same dose of normal saline was intravenously injected in Group C at the same time-point .Patient-controlled intravenous analgesia (PCIA) with fentanyl had implemented in two groups after operation .The levels of interferon-γ(IFN-γ) ,interleukin-4(IL-4) in Th1 and Th2 were measured by flow cytometry before anesthesia(T0) ,2 h(T1) ,12 h(T2) ,24 h(T3) ,48 h(T4)after operation in the two groups .Th1 cell was marked by CD3+ CD8- IFN-γ+ .Th2 cell was marked by CD3+ CD8- IL-4+ .Th1/Th2 was calculated by the level of IFN-γ+ /IL-4+ .The visual analogue scale (VAS) ,Bruggemann comfort scale(BCS) ,dosage of fenta-nyl at each time-point in two groups were compared .Results:VAS scores in group P were significantly lower than those in Group C at T2 ,T3 ,T4 .BCS scores at T1 ,T2 ,T3 in group P were significantly higher than those in Group C .The dosages of fentany at T2 ,T3 in Group P were significantly lower than those in Group C .Compared with T0 ,Th1/Th2 ratios in the two groups at T2 ,T3 ,T4 were lower(P<0 .05) .Th1/Th2 in Group P was higher than those in Group C at T2 ,T3 ,T4(P<0 .05) . Conclusions :Th1/Th2 in patients decreases after radical mastectomy .Preemptive analgesia with parecoxib sodium assisted with fentanyl PCIA can inhibit the shift of the balance of Th1/Th2 towards Th2 type .

Objective To construct an adenoviral vector containing mouse Hes1 gene, observe its expression in the hippocampus of adult mice and build a basis for further investigation of Hes1 gene in adult neurogenesis. Methods The restriction endonuclease was used to digest plasmid pEGFP-mHes1 and pDC316, and then, the products were recovered and connected by T4 DNA ligase and the shuttle plasmid pDC316-mHes1 was constructed which was identified by the method of PCR and EcoRI+HindⅢ digestion. After that, the shuttle plasmid pDC316-mHes1 was cotransfected into 293 cells with the adenovirus skeleton plasmid pBHGlox_E1,3Cre to obtain the produced replication defective recombinant adenovirus Ad5-mHes1. Then, the recombinant adenovirus could be further amplified and purified. The report recombinant adenoviruses were Ad5-EGFP containing enhanced green fluorescent protein (EGFP).Then, Ad5-mHes1 and Ad5-EGFP were stereotactic injected into the hippocampus of the adult C57BL/6 mice and their expressions in the hippocampus were detected. Western blotting was used to detect the Hes1 protein level 7 d after the injection. Fluorescent microscopy was used to observe the expression of EGFP in the hippocampus. Results The experimental results of identification by the methods of PCR and EcoRI+HindⅢ digestion were in accordance with the anticipated results, and the sequences were also the same with mHeslCDS sequences; Hes1 gene was expressed in the hippocampus of both the PBS injection group and the Ad5-mHes1 injection group 7 d after the injection, and the expression of Hes1/GAPDH in Ad5-mHes1 injection hippocampus (0.705 ±0.128) was statistically different as compared with that in PBS injection group (0.363±0.053, P＜0.05). Ad5-EGFP strongly expressed in the granular cell layer and subgranular zone (SGZ) of dentate gyrus. Conclusion The adenoviral vector of mouse Hes1 gene is successfully established and Hes1 gene is expressed in the hippocampus of C57BL/6 adult mice.

ObjectiveTo observe the morphological changes of bone in the progress of chronic fluorosis.MethodsWistar rats were randomly divided into three groups,30 rats in each group:normal control group,experimental group Ⅰ and experimental group Ⅱ according to body weight.Rats in normal control group drank distilled water freely.Experimental group Ⅰ and group Ⅱ drunk distilled water with sodium fluoride preparation of fluorine containing ion 100,150 mg/L solution for six months,respectively.Bone mineral density was detected by X-ray,bone morphological changes were observed under light microscope and bone histomorphometric parameters were calculated using image analysis software.ResultsThe bone mineral density values were different statistically between the three groups after feeding for 2 and 4 months(F =19.79,3.28,all P ＜ 0.05).However no significant difference was found after feeding for 6 months(F =1.80,P ＞ 0.05).The bone mineral density of experimental group Ⅰ (0.20 ± 0.03,0.21 ± 0.03) was significantly higher than that of the normal control group(0.17 ± 0.03,0.20 ± 0.04) after feeding for 2 and 4 months.The bone mineral density of experimental group Ⅱ (0.21 ± 0.02) was lower than that of normal control group(0.22 ± 0.03) after feeding for 6 months.The bone lamella in experimental group Ⅰ was arranged disorderly,the number of osteocytes increased with their nucleus atrophy and the osteoblasts were more than that of control grouo which arranged in layers observed under light microscooy.In exoerimental group Ⅱ,the bone lamella was bent deformation,the number of osteocytes had decreased with their nucleus shrinking or even disappeared and the number of osteoclasts had increased significantly observed under light microscopy.In experimental group Ⅰ,the mean trabecular density [(0.33 ± 0.03)％] increased and the mean trabecular separation,thickness [( 163.57 ± 1.99),(59.26 ± 7.18 ) μm] decreased compared with that of normal control group [(0.31 ± 0.02)％,(186.60 ± 2.90)μm,(86.42 ± 1.48)μm,all P ＜ 0.05].In experimental group Ⅱ,the mean trabecular density[(0.26 ± 0.02)％] decreased,the mean trabecular thickness[(71.42 ± 10.77)μm] reduced compared with that of normal control group[(0.31 ± 0.02)％,(86.42 ± 1.48)μm].ConclusionsExcess fluoride can damage bone tissue.Low doses of fluoride can stimulate osteoblast activity and enhance osteogenesis.The activity of osteoblasts is great than that of osteoclasts.High doses of fluoride can stimulate both osteoblasts and osteoclasts activity,but mainly the activity of osteoclasts,and bone resorption increases.

ObjectiveTo detect the concentration and distribution of fluoride ions in osteoblasts exposed to fluoride in vitro culture,and to provide basic information for studying the effect of fluoride on osteoblast injury.MethodsIn vitro cultured osteoblasts were exposed to 0,5,10,20,40 mg/L fluoride for 3,10,30 d (n =6),respectively.Concentration and distribution of fluoride ions in the cytoplasm and the nucleus of these osteoblasts were determined by nuclear magnetic resonance spectroscopy.Results(①) After cultured for 3 d,fluoride ion content of the bone cytoplasm exposed to different concentrations of fluoride 0,5,10,20,40 mg/L were (0.83 ±0.65),(0.54 ± 0.23),(0.65 ± 0.77),(0.59 ± 0.87),(3.64 ± 1.21 )mg/L,respectively,and the values of exposed to 40 mg/L fluoride group was significantly higher than that of exposed to 0,5 mg/L groups (all P ＜ 0.05).(②)after cultured for 10 d,the composition of the fluoride ion in cytoplasm of exposed to fluoride 10,20,40 mg/L groups were (4.03 ± 1.23),(3.66 ± 0.98),(6.26 ± 2.10)mg/L,respectively,which were higher than that of exposed to 0,5 mg/L groups [(0.78 ± 0.75),(2.69 ± 0.89)mg/L,respectively,all P ＜ 0.05].Of fluoride 20,40 mg/L groups,the composition of the fluoride ion in nucleus were (1.63 ± 1.19),(2.17 ± 1.21 )mg/L,respectively,which were higher than that of 0,5 mg/L groups[(0.65 ± 0.46),(1.57 ± 0.33) mg/L,all P ＜ 0.05].(③)After cultured for 30 d,of the exposed to fluoride 10,20,40 mg/L groups,the composition of the fluoride ion in cytoplasm were (3.99 ± 0.84),(4.33 ± 1.67),(5.80 ± 1.38)mg/L,respectively,which were higher than that of 0,5 mg/L groups[(0.88 ± 0.44),(2.84 ± 0.43)mg/L,all P ＜ 0.05].The composition of the fluoride ion in nucleus of the fluoride 20,40 mg/Lgroups were (3.33 ± 1.46),(3.53 ± 1.22)mg/L,respectively,which were significantly higher than that of 0,5mg/L groups [(0.70 ± 0.66),(1.99 ± 0.76)mg/L,all P ＜ 0.05].ConclusionsWhen osteoblasts are exposed to fluoride environment,fluoride ions enter into the osteoblasts quickly,and quickly accumulate in the nucleus,showing a special affinity between fluoride and bone tissue.Intracellular fluoride ions increase with the increase of contact time and exposure dose.

Objective:To investigate the predictive value of serumal retinol-binding protein 4(RBP4) level fro fetal macrosomia of non-diabetic pregnant women .Methods :The serumal levels of RBP4 of 500 non-diabetic pregnant women at 12 week ,20 week and 24 week of pregnancy were measured by immune projection turbidimetric method .Fetal macrosomia was defined as birth weight≥4000 g .The cut-off value ,sensitivity and specificity were calculated with receiver operating characteristic (ROC) curve .Results:Of the 500 non-diabetic pregnant women ,30 cases(6% ) got fetal macrosomia .The ROC curve showed that the predictive cut-off values of RBP4 at 12 week ,20 week and 24 week of pregnancy were 61 .0 mg/L ,50 .5 mg/L and 52 .5 mg/L , respectively ;the predictive sensitivity and specificity at 12 week ,20 week and 24 week of pregnancy were 42 .9% and 94 .5% , 70 .0% and 69 .5% ,76 .9% and 73 .2% ,respectively .The predictive cut-off value of RBP4 no later than 24 week of pregnancy was 51 .5 mg/L ;the predictive sensitivity and specificity were 61 .8% and 69 .5% .There was significant difference(P<0 .05) between the serumal level of RBP4 at 24 week of pregnancy in group fetal macrosomia and that in group nonfetal macrosomia . Conclusions :The predictive sensitivity of RBP4 increases in accordance with the increase of serumal level of RBP 4 .The serumal level of RBP4 of non-diabetic pregnant women at 24 week of pregnancy may have higher sensitivity and specificity in the predic-tion of fetal macrosomia .If the serumal level of RBP4 no later than 24 week of pregnancy is beyond 50 mg/L ,then the risk of fetal macrosomia will be higher .