A case of several ambushed bicuspoid supernumerary teeth in the premolar area was reported.The size,shape and the ratio of crown to root of the supernumerary teeth are similar to those of the premolar.The abnormity of tooth germ division and proliferation may lead the supernumerary teeth.

Aim: To establish an analytical method for the simultaneous determination of norfloxacin,ofloxacin,pefloxacin,ciprofloxacin,lomefloxacin,danofloxacin,enrofloxacin,sarafloxacin and difloxacin in eggs using solid-phase extraction-LC-MS/MS.Methods: Egg samples were deproteinized with acetonitrile,followed by defatting with hexane.Then the samples were processed by solid-phase extraction and analyzed by LC-MS/MS using an electrospray source.The separation was carried out on a Shimadzu Shim-pack VP-ODS C_(18) column,with a mobile phase consisting of acetonitrile-0.1% formic acid(13: 87).Results: The validated method was proved to be of high specificity,accuracy and sensitivity.Conclusion: The established method is suitable for the routine residual monitoring of fluoroquinolones.

Objective To observe the effect of Sishen Pill on ICAM-1 mRNA and protein expression of colonic mucosa in rats with ulcerative colitis (UC), and identify its mechanism. Methods Taolly 40 SPF Wistar rats were randomly divided into blank group, model group, Pill group and SASP group. Except the blank group, UC model was prepared with TNBS/ethanol enema. Pill group was given Sishen Pill 5 g/kg, and SASP group was given SASP 0.3 g/kg by gavage, blank group and model group was given the same volume physiological saline for three weeks. Morphological injury of colonic mucosa was observed and scored. ICAM-1 gene and protein expression were detected by RT-PCR and immunohistochemical method. Results Inflammation and ulceration were found on the colonic mucous membrane of rats in the model group. The expression of ICAM-1 gene and protein of colonic tissues of rats in the model group increased compared with that of the blank group (P<0.01). Compared with the model group, the expression of ICAM-1 in Pill group decreased (P<0.05, P<0.01). Conclusion Sishen Pill can decrease the expression of ICAM-1 mRNA and ICAM-1 protein, inhibit the infiltration of inflammation cells, prevent and reduce colon tissue damage, and play a vital role in the treatment of UC.

Objective:To investigate the effect and mechanism of lysine acetyltransferase 5(KAT5) on the radio-sensitivity of anaplastic thyroid carcinoma (ATC).Methods:The expression levels of endogenous KAT5 in ATC and normal thyroid cells were detected by Western blot and qRT-PCR. The effect of KAT5 specific inhibitor NU9056 on the radio-sensitivity of human ATC cells and normal thyroid cells was evaluated by colony formation assay. TCGA database, JASPAR database, along with Western blot, microRNA sequencing, qRT-PCR and dual-luciferase reporter assay were conducted to unravel the underlying mechanism.Results:The expression of endogenous KAT5 at the protein and mRNA levels in human ATC cells was significantly higher than that in normal thyroid cells. NU9056 could significantly enhance the radiosensitivity of human ATC cells to 8505C and CAL-62, whereas showed no sensitization effect on normal thyroid cell Nthy-ori 3-1. Knockdown of KAT5 and NU9056 both down-regulated the expression level of miR-210 in the TC cells, while NU9056 decreased the expression level of transcription factor c-Myc. The putative binding sites of c-Myc in the miR-210 promoter region were predicted, and transfection of c-Myc plasmid significantly enhanced the luciferase activity of miR-210 promoter. Elevated miR-210 level was associated with worse survival of patients with thyroid carcinoma. Down-regulated expression of miR-210 decreased the TET2 mRNA level, while inhibition of miR-210 increased the TET2 mRNA level.Conclusion:The aberrantly-activated KAT5/miR-210/TET2 pathway probably causes the radioresistance of ATC, becoming a novel sensitizing target for ATC radiotherapy in clinical practice.

Objective To evaluate the relationship between positive thyroid autoantibody and risk of preterm birth.Methods Literature search was done in PubMed,Embase,China Academic Journal Network Publishing Database,Wanfang Medical Database and China Biology Medicine disc databases from January 1st,1989 to January 26th,2012.Criteria for inclusion included:(1) Prospective cohort study; (2) The exposure was positive thyroid autoantibody and outcome was preterm birth; (3) The enrolled subjects were pregnant women without cardiovascular or rheumatic disease; (4) Relative risk (RR) and its 95％ confidence interval (95％CI) of preterm birth were provided in the study.Meta-analysis was performed by Stata 12.0.The relationship between positive thyroid autoantibody and risk of preterm birth was evaluated by RR and 95％ CI.Results Ten cohort studies were enrolled.One thousand six hundred and fifty seven cases of preterm birth occurred among 25 081 pregnant women.Heterogeneity among the 10 studies was found in meta-analysis (I2 =79.2％,P＜0.01).The risk of preterm birth in pregnant women with positive thyroid autoantibody was higher than those in control group by random effects analysis (RR=1.61,95％CI:1.18-2.20,P＜0.05).Subgroup analysis was further performed.In five studies,the cases of control group were pregnant women with normal thyroid function; heterogeneity was not found in these five studies (I2=39.1％,P=0.160); and RR of the risk of preterm birth was 2.55 in pregnant women with positive thyroid autoantibody (95 ％ CI:2.04 3.19,P＜0.01).In the other five studies,the cases of controlgroup were pregnant women who had not been ruled out the possibility of thyroid dysfunction;heterogeneity was not found in these five studies either (I2 =0.0％,P =0.970); and RR was 1.18(95 ％ CI:1.01-1.37,P＜0.05).After excluding two low-quality studies,RR of the risk of preterm birth was 1.72 in pregnant women with positive thyroid autoantibody (95％CI:1.18 2.53,P＜0.05).The funnel plots presented symmetrical graphics,indicating that there was no publication bias.Conclusions Positive thyroid autoantibody in pregnant women is a risk factor of preterm birth.

Background Graft rejection is a primary cause of corneal transplantation failure,especially in high-risk keratoplasty.How to extend the survival time of graft is a problem to be solved.Objective This study was to investigate the influence of immune tolerance on high-risk rat keratoplasty induced by donor bone marrowderived immature dendritic cells (imDCs) transfected by chemokine receptor 7 (CCR7) recombinant adenovirus (Ad).Methods Bone marrow-derived imDCs were isolated and cultured from femur marrow of one male Wistar donor rat.The cells were transfected using recombinant Ad vector with rat CCR7 gene and resuspended in 500 μl PBS containing 1％ fetal bovine serum with the cells 1 × 107.High-risk corneal transplantaion models were established using monolateral corneal alkali-burn method in 60 SD rat recipients, and then allograft keratoplasty was performed with the 30 Wistar rats as donors.The models were randomized into the PBS group,imDCs group,imDCs with blank Ad vector group and imDCs with Ad-CCR7 group following the corresponding solution injection via caudal vein on preoperative day 7 and postoperative day 3 respectively.The survival time of graft was evaluated under the slit lamp microscope once per day.On the 14th day after operation, corneal sections were prepared from 6 eyes of each group for the pathological examination,and the relative expression levels of T helper cell 1 (Th1)-related factors,interleukin-2 (IL-2) mRNA and interferon-γ(IFN-γ) mRNA,as well as Th2-related factors, IL-4 mRNA and IL-10 mRNA, were detected by reverse transcription PCR (RT-PCR).The use and care of animals complied with the ARVO statement Results The mean survival time of grafts was (10.44±1.88) , (16.00±2.18) , (15.11±2.03) and(23.67±2.83) days in the PBS group,imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group, respectively, with a significant difference among the 4 groups (F =53.005, P =0.000).Compared with the PBS group, the mean survival time of grafts was considerably extended in the imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group (t=5.220,4.385,12.423 ,all at P=0.000) ,and a remarkble prolongation of graft survival duration was seen in the imDCs with Ad-CCR7 group in comparison with the imDCs group and the imDCs with blank Ad group (t =7.204,8.039,both at P=0.000).The relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were significantly lower,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were significantly higher in the imDCs group,imDCs with blank Ad group and the imDCs with Ad-CCR7 group than those in the PBS group (all at P＜0.05).Compared with the imDCs group and the imDCs with blank Ad group, the relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were remarkably delined,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were remarkably elevated in the imDCs with Ad-CCR7 group (all at P =0.000).Conclusions Application of imDCs transfected with CCR7 recombinant Ad via vena caudalis can prolong the survival time of grafts after keratoplasty of SD rats probably by affecting the balance of Th1/Th2 cytokines.

Background and purpose:miR-196a2 functions as an oncogene during tumor initiation and pro-gression. The up-regulation promotes tumor cell proliferation, invasion and metastasis. Therefore, it is promising to be an important tumor biomarker. The aim of this study was to investigate whether rs11614913, a gene polymorphic site ofmiR-196a2, is associated with the risk of leukemia.Methods:A case-control analysis was employed. Bone marrow or periph-eral blood was collected from 210 leukemia patients diagnosed from Jan. 2009 to Jul. 2015 in Yantaishan Hospital (case group) as well as 250 healthy people who were physically examined during the same period (control group). Polymerase chain reaction-restriction fragment length polymorphism (PCR-PFLP) was used to detect the genotype of rs11614913. Application test was used to compare the difference in the frequency of each genotype between case group and control group. The odds ratio (OR) of SNP allelic genes was calculated using logistic regression analysis and 95%CI represented the risk of leukemia for each genotype.Results:The distribution differences in the frequency of T/T, C/C, C/T genotype of miR-196a2 rs11614913 between case group and control group were statistically significant (P<0.05). The risk of leukemia for individuals who carried mutant homozygous C/C was 2.661-fold higher than those carried wild-type homozygous T/T, and the difference was statistically significant (P<0.05).Conclusion:ThemiR-196a2 gene polymorphic site rs11614913 was associated with the risk of leukemia. Mutant homozygous C/C or C allelic gene carrying was probably a risk factor for leukemia.

Objective To study the cooperative method of goat's cruciate ligament reconstruction by bone-tendon autograft with interference fixation. Methods A customized reamer was used to make the bottleneck-like femoral tunnel, and the patellar tendon-tibial tuberosity autograft was harvested. Make sufficient preparation for this operation. Results The operation was successful, the laboratory animals all survived without any postoperative infection. Conclusion Scientific and careful operative-cooperation can guarantee the successful operation.

Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2)/ antioxidant response element (ARE) signaling pathway in reduction of myocardial ischemia-reperfusion (I/R) injury by ischemic preconditioning in the rats.Methods Healthy male Sprague-Dawley rats,weighing 250-300 g,aged 4-6 months,were used in the study.Their hearts were excised,and retrogradely perfused with K-H solution at 37 ℃ in a Langendorff apparatus.Forty isolated hearts were randomly divided into 4 groups (n=10 each) using a random number table:control group (group C),I/R group,ischemic preconditioning group (group IPC),and ischemic preconditioning +Nrf2/ARE signaling pathway blocker luteolin group (group IPC+L).After 20 min of equilibration,the hearts were continuously perfused for 100 min in group C.After 20 min of equilibration,the hearts were subjected to 40 min ischemia at 32 ℃ followed by 60 min of reperfusion in group I/R.In group IPC,ischemic preconditioning was induced by 6 cycles of 10 s ischemia followed by 10 s reperfusion starting from the time point immediately after 20 min of equilibration,and then the hearts were subjected to 40 min ischemia at 32 ℃ followed by 58 min of reperfusion.In group IPC+L,after 20 min of equilibration,the hearts were perfused with K-H solution containing lueolin 50 μmol/L for 3 min before ischemia,and the other treatments were similar to those previously described in group IPC.Left ventricular developed pressure (LVDP),left ventricular end-diastolic pressure (LVDEP),heart rate (HR),and the maximum rate of increase of left ventricular pressure (+dp/dtmax) were recorded at the end of equilibration and reperfusion.At the end of reperfusion,left ventricular myocardial tissues were obtained for examination of the ultrastructure of myocardial cells and for determination of the expression of Nrf2,heme oxygenase-1 (HO-1),quinone oxidoreductase 1 (NQO1),and superoxide dismutase 1 (SOD1) mRNA and protein (by real-time polymerase chain reaction and Western blot,respectively).Results Compared with group C,the HR,+ dp/dtmax and LVDP were significantly decreased,and LVEDP was significantly increased at the end of reperfusion in I/R and IPC+L groups,and the expression of Nrf2,HO-1,NQO1 and SOD1 mRNA and protein was significantly up-regulated in I/R,IPC and IPC+L groups (P＜0.05).Compared with group I/R,the HR,+dp/dtmax and LVDP were significantly increased,and LVEDP was significantly decreased at the end of reperfusion,the expression of Nrf2,HO-1,NQO1 and SOD1 mRNA and protein was significantly up-regulated (P＜0.05),and the pathological changes were significantly attenuated in group IPC,and no significant change was found in the parameters mentioned above in group IPC+L (P＞0.05).Compared with group IPC,the HR,+dp/dt and LVDP were significantly decreased,and LVEDP was significantly increased at the end of reperfusion,and the expression of HO-1,NQO1,SOD1 mRNA and protein was significantly down-regulated (P＜ 0.05),no significant change was found in Nrf2 mRNA and protein expression (P＞0.05),and the pathological changes were significantly aggravated in group IPC + L.Conclusion Ischemic preconditioning reduces myocardial I/R injury through activating Nrf2/ARE signaling pathway in the rats.

Objective To investigate the humoral and cellular immune responses in patients with acute brucellosis, and evaluate dynamic changes of regulatory T-lymphocytes (Foxp3+ Treg) in the peripheral blood of patients during treatment, in order to clarify the relationship between immunosuppression and the therapeutic effect in human brucellosis.Methods Sixty-five patients with brucellosis hospitalized at the Third Department of Infectious Diseases, Heilongjiang Agriculture and Reclamation Bureau General Hospital between July 2015 and November 2015 were included.Twenty-eight patients were treated with conventional therapy (group A: patients received 3 courses of treatment.Each lasted for 20 days with one-week interval), and 37 patients were treated with conventional therapy in combination with immunopotentiator (group B).Thirty healthy volunteers were enrolled as the controlled group.The ratio of CD3+CD4+ Foxp3+ Treg cells in the peripheral blood of brucellosis patients were measured by flow cytometry (FCM) at the end of each course of treatment.Data in accordance with normal distribution were described as mean±standard deviation.Comparison between two groups was done by two sample t test.Comparison among multiple groups was performed by analysis of variance and SNK test.Data that did not fit the normal distribution were analyzed by multiple-sample nonparametric test.Results After the first (20 d), second (50 d) and third course of treatment (80 d), the ratios of Foxp3+Treg in the peripheral blood of 65 acute brucellosis patients were 2.83%, 3.77% and 4.03%, respectively, which were all significantly higher than control group (1.69%;t=5.97, 9.05 and 5.66, respectively, all P<0.01).At the end of the first course of treatment, the ratios of Foxp3+Treg in group A and B showed no statistically difference (t=0.33, P>0.05), while those were both higher than control group (t=7.09 and 4.94, respectively;both P<0.01).At the end of the second course, the ratio of Foxp3+ Treg in group B was higher than group A (t=2.22, P<0.01), and both of them were higher than control group (t=10.79 and 7.25, respectively;both P<0.01).At the end of treatment, Foxp3+ Treg in group A was also significantly higher than the other two groups (t=6.02 and 6.45, respectively;both P<0.01).Conclusions In patients with acute brucellosis treated with the standard antibiosis treatment in combination with immunopotentiator, the ratio of Foxp3+Tregs significantly increases and maintains at a high level, which suggests that extra immunopotentiator may be not helpful for the treatment of brucellosis at the very early stage.