Purpose To investigate the correlation between the methylation level at CpG sites of CXCL17 and the clinicopathological parameters of papillary thyroid carcinoma(PTC).Methods samples from 186 cases of PTC and 191 cases of benign thyroid nodule(BTN)were collected.Methylation levels of CXCL17 were semi-quantitatively as-sessed using mass spectrometry.Logistic regression analysis,which adjusted for age,gender and related hormones,was conducted to evaluate the correlation between CXCL17 methylation and PTC,and calculate the odds ratios(ORs)and 95％confidence intervals(CIs).Results Hypomethylation level of CXCL17_CpG_1.2 was significantly associat-ed with PTC(OR=1.36,95％CI:1.16-1.60,P＜0.001)and early stage of PTC patients(Stage Ⅰ,OR=1.41,95％CI:1.19-1.67,P＜0.001).Gender-based hierarchical management analysis showed that decreased methyla-tion level of CXCL17_CpG_1.2 was significantly associated with female PTC patients(OR=1.39,95％CI:1.15-1.67,P＜0.001).In subgroups stratified by age(＜50 and≥50 years old),hypomethylation at CXCL17_CpG_1.2 was significantly associated with PTC,with a stronger association in the younger subgroup(＜50 years old:OR=1.42,95％CI:1.14-1.77,P＜0.01;≥ 50 years old:OR=1.30,95％CI:1.03-1.64,P＜0.05).Conclusion There was a significant difference in CXCL17 methylation levels between benign and malignant thyroid tumors.It was showed that hypomethylation of CXCL17 is closely associated with PTC,particularly in young women patient.Thus,CXCL17 methylation may serve as a biomarker for accurate differential diagnosis of thyroid nodule.

Objective To investigate the effects and mechanisms of metformin-induced metabolic intervention on the proliferation and differentiation of myoblasts.Methods C2C12 myoblasts cultured in growth or differentiation media were treated with 1 mmol/L metformin,with DMSO as the control.Cells in growth media were treated for 48 hours,with samples collected every 12 hours for metabolomic analysis,protein expression,mitochondrial function,and proliferation assays.Cells in differentiation media were induced to differentiate for 72 hours,and myotube formation was assessed via embryonic myosin heavy chain immunofluorescence staining.Results Metformin treatment increased intracellular levels of metabolites such as lactate,pyruvate,and adenosine monophosphate,while reducing adenosine triphosphate and thiamine pyrophosphate.Phosphorylation of AMP-activated protein kinase(AMPK)and related protein expression were significantly upregulated(P<0.05).Mitochondrial biogenesis was enhanced,and membrane potential markedly decreased(P<0.05).The proportion of S-phase cells and EdU-positive cells gradually increased(P<0.01),though overall proliferation rate slowed.During differentiation,the myoblast fusion index significantly decreased(P<0.01).Conclusion Metformin induces metabolic reprogramming in myoblasts and suppresses their proliferation and differentiation,with activation of AMPK-related signaling pathways serving as a key molecular mechanism.

Purpose To investigate the correlation between the methylation level at CpG sites of CXCL17 and the clinicopathological parameters of papillary thyroid carcinoma(PTC).Methods samples from 186 cases of PTC and 191 cases of benign thyroid nodule(BTN)were collected.Methylation levels of CXCL17 were semi-quantitatively as-sessed using mass spectrometry.Logistic regression analysis,which adjusted for age,gender and related hormones,was conducted to evaluate the correlation between CXCL17 methylation and PTC,and calculate the odds ratios(ORs)and 95％confidence intervals(CIs).Results Hypomethylation level of CXCL17_CpG_1.2 was significantly associat-ed with PTC(OR=1.36,95％CI:1.16-1.60,P＜0.001)and early stage of PTC patients(Stage Ⅰ,OR=1.41,95％CI:1.19-1.67,P＜0.001).Gender-based hierarchical management analysis showed that decreased methyla-tion level of CXCL17_CpG_1.2 was significantly associated with female PTC patients(OR=1.39,95％CI:1.15-1.67,P＜0.001).In subgroups stratified by age(＜50 and≥50 years old),hypomethylation at CXCL17_CpG_1.2 was significantly associated with PTC,with a stronger association in the younger subgroup(＜50 years old:OR=1.42,95％CI:1.14-1.77,P＜0.01;≥ 50 years old:OR=1.30,95％CI:1.03-1.64,P＜0.05).Conclusion There was a significant difference in CXCL17 methylation levels between benign and malignant thyroid tumors.It was showed that hypomethylation of CXCL17 is closely associated with PTC,particularly in young women patient.Thus,CXCL17 methylation may serve as a biomarker for accurate differential diagnosis of thyroid nodule.

Objective : To explore the relationship between the methylation level of RABL6 in peripheral blood and early lung cancer (LC) with a case-control study in the Chinese population． Methods : The methylation levels of 7 CpG sites in RABL6 gene in peripheral blood of samples from 275 LC patients (81．5% at stage I) ，and age- and gender-matched 185 benign lung nodule cases and 267 matched healthy controls were measured by matrix-assisted laser desorption ionization time-of-flight mass spectrometry．Multinomial Logistic regression adjusted for covariates was used to analyze the association between the RABL6 methylation and LC．Mann-Whitney U test was applied for the comparisons of RABL6 methylation levels between clinical characteristics subgroups of LC. Results : Compared to the healthy controls，the methylation of RABL6 _CpG_ 17 was inversely associated with LC in females ( per - 10% methylation : OR = 2. 47，95% CI = 1. 19－5. 13，P = 0. 016) ，but positively associated with LC in males (per - 10% methylation : OR = 0. 52，95% CI = 0. 29－0. 94，P = 0. 030) ．In addition，hypermethylation of RABL6_CpG _2 and RABL6_CpG_5 was significantly associated with LC in the subjects older than 55 years (for RABL6_CpG_ 2 : per －10% methylation : OR = 0. 77，95% CI = 0. 60－0. 99，P = 0. 038 ; for RABL6_CpG_5 : OR = 0. 58，95% CI = 0. 34－0. 97，P = 0. 038) ． Conclusion The study reveals an association between peripheral blood-based RABL6 methylation levels and early LC，providing a new clue for developing peripheral blood-based DNA methylation as a potential marker for the evaluation of LC risk.

Objective To observe the impairment effect of retinol binding protein 4(RBP4) on neurocognitive function in diabetic nephropathy(DN) patients with silent cerebral infarction(SCI) and to explore its mechanism.Methods Sixty patients with newly diagnosed DN and 30 healthy volunteers were selected as the study subjects and the DN cases were divided into the complicating SCI group(SCI,n=30) and non-complicating SCI group(NSCI,n=30) according to the imaging results.The degrees of neurological function deficit and Montreal cognitive assessment(MoCA) were evaluated.Serum RBP4 level was determined by ELISA and expressions of Lp-PLA2 and C-X-C chemokine receptor type 4(CXCR4) were determined by Western blot.Results Compared with the NSCI group,the neurocognitive function in the SCI group was subsided,the expression levels of RBP4,Lp-PLA2 and CXCR4 were increased(P＜0.05).The RBP4 level was positively correlated with the neurocognitive function impairment in SCI patients,moreover,there existed a regression correlation between them.Conclusion Serum RBP4 may serve as the predictive factor of DN complicating SCI and is positively correlated with neurocognitive dysfunction.Lp-PLA2/CXCR4 pathway activation may be one of its pathogenesis.

Objective To observe the effect and efficacy of DPP-4 inhibitor saxagliptin on nonalcoholic fatty liver disease (NAFLD) in patients with type 2 diabetes mellitus.Methods Forty patients with newly diagnosed with type 2 diabetes complicated with NAFLD were divided into two groups:the saxagliptin group and the metformin group.On the basis of dietary and exercise therapies,the saxagliptin group received 0.005 g of saxagliptin daily and the metformin treatment group received 0.85-1.70 g of metformin daily.The levels of glycosylated hemoglobin(HbA1 c),triglyceride (TG),blood pressure (BP),alanine aminotransferase (ALT),liver CT and liver/spleen CT were observed before and 3 months after treatment.Results After treatment for 3 months,the TG,ALT,liver CT and liver / spleen CT of the metformin group and the saxagliptin group were significantly higher than those before treatment(P＜0.05).There was no significant difference in the TG,ALT and liver CT values between the two groups(P＞ 0.05).Conclusion In the state of diabetes mellitus complicate with nonalcoholic fatty liver,saxagliptin has a certain effect on relieving fatty liver.

Objective To discuss the value of color Doppler ultrasound in the diagnosis and differential diagnosis of ovarian granulosa cell tumor (OGCT) . Methods Fourteen cases were diagnosed as OGCT by pathology postoperatively had been examined by color Doppler ultrasound check-up before the surgery . In image appearance ,the tumors were divided into cystic mass ,solid mass or mixed cystic-solid mass respectively according to their own pathological features . Additionally ,the blood flowing signal of OGCT was labeled from 0 to 3 grade with Adler standard based on the blood flowing imaging observed by color Doppler flow imaging ( CDFI) . The CDFI results were compared with the pathological diagnosis . Results Ultrasound image appearance of OGCT varied in the 14 cases .Two of them could be characterized as cystic mass ,5 solid mass and 7 mixed cystic-solid mass . The mixed cystic-solid OGCT contained solid mass in multiple cystic degeneration area . There was a significant difference in the diameter among three kinds of OGCT ( P = 0 .0006) . The more visible cystic lesion indicated the longer diameter of OGCT . According to the CDFI ,4 ,5 ,3 and 2 cases were 0 ,1 ,2 ,3 grade respectively in Adler standard . No significant difference was found among the four standards groups ( P =0 .056) . The localization diagnosis accordance rate of the Doppler imaging in OGCT was 71 .4％ (10/14) and the qualitative diagnosis accordance rate was 35 .7％ (5/14) . Conclusions The color Doppler ultrasonography can be the first-line preoperative imaging investigation in the diagnosis and differential diagnosis of OGCT ,as well as in the long-term following after surgery . It can provide more details and evidences for the treatment and prognosis assessment of OGCT .

Objective To explore the way of contrast-enhanced ultrasound(CEUS)and quantitative analysis technology to assess testicular microvascular ischemia-reperfusion inj ury.Methods Twenty-four healthy adult rabbits were randomly divided into the control group and the microvascular mild,moderate, serve ischemic inj ury group (group Ⅰ,including group Ⅰ1 ,Ⅰ2 ,Ⅰ3 ).The spermatic cord of the control group were threaded but not ligated.The group Ⅰ1 ,Ⅰ2 ,Ⅰ3 were reperfused 2 h after 2 h,4 h,6 h complete testicular ischemia respectively.The ultrasound contrast angiographic parameters of each group were analyzed by the quantitative analysis techniques,including peak intensity(PI),time to peak(TP),slope,mean transmit time(MTT),DT/2 and area.The levels of plasma vWF,NO and ET-1 were tested after the CEUS. The pathological changes of the ipsilateral testicular were observed by the HE and immunohistochemical staining.Statistical analysis was performed by SPSS 13.0 software.Results Preoperativeiy,each parameter of the experimental group and the control group showed no statistical significance (P > 0.05 ). Postoperatively,the PI,slope,area of the group Ⅰ1 increased significantly,while the TP,MTT were prolonged(P <0.05).Contrast time-intensity curve was significantly higher peak and retrusion.The groupⅠ2's PI,slope,area increased further,DT/2 was significantly prolonged,but TP significantly reduced (P <0.05).The contrast curve significantly increased,forward and decreased slowly.The contrast curves of the group Ⅰ3 was approximate a straight line.The vWF,NO,ET-1 levels of each experimental group were significantly higher than those in control group (P <0.05),among them,the levels of group Ⅰ2 were the highest (P <0.05 ).With the aggravation of ischemia,the testicular capillaries expansion,erythrocyte sedimentation,leakage,infiltration of inflammatory cells,MMP-9 expression in vascular basement membrane gradually increased.Conclusions The CEUS and quantitative analysis techniques can effectively evaluate testicular microvascular inj ury.

Objective To probe the value of color Doppler flow imaging(CDFI) in diagnosis of borderline ovarian epithelial tumors (BOET).Methods Thirty-six cases of BOET,139 cases of ovarial cystadenoma and 66 cases of ovarial cystadenocarcinoma were diagnosed pathologically.All cases were preoperatively detected by CDFI.Tumor sign CA125 was also detected.The size,boundary,internal echo and internal flow distribution were observed.The results of CDFI of different diseases were contrasted.Results Ultrasonic diagnosis rate was 75 ％ (27/36) in BOET,90.6 ％ (126/139) in ovarial cystadenoma and 92.4 ％ (61/66) in ovarial cystadenocarcinoma.Positive rate of CA125 of ovarial cystadenocarcinoma was obviously higher than those of BOET and ovarial cystadenoma.Positive rate of CA125 of ovarial cystadenocarcinoma containing solid was especially higher than that of BOET containing solid.Ovarial cystadenoma mainly showed single cavity,few or without solid as well as hematoasthenia or asanguineous.BOET and ovarial cystadenocarcinoma mainly showed multi-cavity,more solid as well as plentiful blood flow.Blood flow of BOET distributed regularly comparing with ovarial cystadenocarcinoma.Conclusions Characteristics of CDFI combining results of CA125 can provide powerful reference for diagnosise and differentiate diagnosis of BOET and more information diagnostic information for clinic to chosing treatment scheme and postoperative long-term follow-up.

Objective To study the impact of ultrasound contrast agent with different doses or imaging modes on rabbit spermatogenic cell in the diagnosis of radiation dose.Methods Thirty-three New Zealand white rabbits were randomly divided into 3 groups.In group Ⅰ ,three minutes gray scale ultrasound radiation was performed for three rabbits.In group Ⅱ, SonoVue was bolus injected from ear vein for eighteen rabbits.The rabbits in this group were divided into 6 sub-group according to different doses and different sampling times, the single injections of SonoVue with dosage of 0.1 ml/kg(A1, A2) or 1 ml/kg (C1 ,C2) and repeated injections with dosage of 0.1 ml/kg interval 15 min (B1 ,B2) were taken.In group Ⅲ ,twelve rabbits were bolus injected with SonoVue and then the agent bubbles were blasted when the SonoVue suffused the whole testis.The rabbits in this group were divided into 4 sub-groups according to different doses and different sampling times, and the dosages of SonoVue were 0.1 ml/kg(D1, D2) and 1 ml/kg(E1,E2).Testis were drew immediately or 24 hours later for the observation under light microscope and transmission electron microscope.Results Apoptotic cells in group Ⅲ were more than those in group Ⅰ and Ⅱ ( P ＜0.05) ,while no significant difference of the total number of apoptotic cells between group Ⅰ and Ⅱ ( P ＞0.05).The appearances of focal ultra-structural damage were observed by transmission electron microscope in group C1, such as intercellular space widening and mitochondria swelling, and no distinct damage were observed in the other groups.The appearances of intercellular space widening, mitochondria swelling and cellular edema were observed by transmission electron microscope in group Ⅲ, and spotty necrosisetc was also observed in group D1 and E1.Conclusions Large dose of contrast agent could cause germ cells slight instantaneous effects, while conventional dose had no effect.Contrast agent blasting may cause germ cells unrecoverable damage.