Objective To investigate the expression of Yes‐associated protein(YAP) in human papillary thyroid cancer and its influence on cell growth .Methods The samples in 57 cases of papillary thyroid cancer treated by operation resection in the gen‐eral surgery department of this hospital and the matched tumor‐adjacent tissues were collected .All the cases were definitely diag‐nosed by the pathology examination .The expression of YAP protein in the cancer tissue and corresponding tumor‐adjacent tissue were determined by the immunohistochemistry(IHC)staining .The relationship between the YAP protein and the clinicopathological data was statistically analyzed .siRNA was used to silence the expression of YAP in B‐CPAP cells ,MTT and flow cytometry were used to measure the proliferation and apoptosis changes .Results The expression level of YAP was markedly higher in papillary thyroid cancer tissues than in tumor adjacent tissues (P<0 .05);moreover the expression of YAP protein was positively correlated with the tumor size and TNM stage (P<0 .05) .Silencing YAP gene could significantly inhibit the cell proliferation ability and pro‐mote cell apoptosis(P<0 .05) .Conclusion YAP is highly expressed in papillary thyroid cancer tissues and is related with its ad‐verse clinicopathological characteristics ,down‐regulating YAP gene can significantly inhibit the cell growth .

Objective To investigate the relationship between the expression of CC chemokine ligand 5(CCL5)and S100 calcium binding protein A4 (S100A4)protein in breast cancer tissues with clinicopathological features and prognosis.Methods The expression of CCL5 and S100A4 in 40 cases of normal breast tissues and 120 cases of breast cancer were detected by immunohistochemistry,and the relationship between the degree of expres-sion and the clinicopathological features and prognosis of breast cancer was analyzed.Results The expression posi-tive rates of CCL5 and S100A4 in breast cancer tissues were 56.67% and 62.50% respectively,which were not expressed in normal breast tissues,and the differences were statistically significant (χ2CCL5 =39.403,P <0.01;χ2S100A4 =47.062,P <0.01).The expression of CCL5 in breast cancer tissues was statistically correlated with clinical staging(χ2 =10.141,P <0.01 ),pathological type (χ2 =5.769,P =0.017)and lymph node metastasis (χ2 =34.178,P <0.01),but not correlated with patients'age,tumor size,pathological grading(all P >0.05 ).And the expression of S100A4 was statistically correlated with clinical staging(χ2 =44.311,P <0.01)and lymph node metas-tasis(χ2 =14.843,P <0.01 ),but not correlated with patients'age,tumor size,pathological type and pathological grading(all P >0.05).The expression of CCL5 and S100A4 in breast cancer was positively correlated(r =0.301, P <0.01).CCL5 was positively correlated with the recurrence of breast cancer(OR =6.270,P <0.01),and S100A4 was not correlated with the recurrence of breast cancer(OR =1.103,P =0.768).Survival analysis showed that the disease -free survival time of patients with positive CCL5 expression was significantly shorter than the patients with negative CCL5 expression(χ2 =11.851,P <0.01 ),and the disease -free survival time of patients with positive S100A4 expression was significantly shorter than the patients with negative S100A4 expression(χ2 =5.433,P =0.021).The joint detection showed that the disease -free survival time in CCL5(+)+S100A4(+)group was sig-nificantly lower than that of CCL5(+)or S100A4(+)group(χ2 =15.341,P <0.01)and CCL5 (-)+S100A4 (-)group(χ2 =15.341,P <0.01).Conclusion The expression of CCL5 and S100A4 in breast cancer can reflect the metastasis and staging of breast cancer,which can be used to judge the clinical pathological characteristics and prognosis of breast cancer.

Objective To identify the arteriosclerosis (AS)changes in peripheral artery and abdominal aorta of patients with familial hypercholesterolemia(FH) during follow-up.Methods Seventeen patients of 6 FH [5 male and 12 female with average age of (16.12?6.65) years old],along with 17 subjects of matching sexes and ages with normal blood cholesterol as healthy control group,underwent examination by color doppler ultrasound,and changes of intima-media thickness (IMT) in peripheral artery and abdominal aorta,severity of stenosis,morphology,and function were observed.Results For 17 patients of FH,the total cholesterol(TC),low density lipoprotein cholesterol(LDL-C) in serum were higher significantly and high density lipoprotein cholesterol (HDL-C) was lower significantly than those in healthy control group(Pa0.05).The IMT of carotid aorta,subclavicular aorta,common abdominal artery,and common femoral artery in 13 patients were showing various degrees of increase,yielding an average of 2.9 mm.As patients aged,this phenomenon was reported to be more profound in common abdominal aorta and femoral artery.Conclusions Patients of FH show AS lesion in early stage and it worsened as they aged,from carotid arteries to common abdominal aorta and femoral artery.Color doppler ultrasound can be a non-invasive examination for monitoring the progress of AS in blood vessels in patients of diagnosed FH.

· AIM: To explore the effects of C3F8 tamponade on hypotony on or after primary operation and the prognosis of severe ocular trauma.· METHODS: Twenty-six cases (26 eyes) of severe ocular trauma were treated with pure C3F8 tamponade on or after primary operation. IOP was observed, and the curative effect of C3F8 tamponade was observed on secondary operation with prognosis evaluated.· RESULTS: Hypotony improved in 23 eyes postoperatively,in which 18 eyes with edematous and cloudy cornea, 15 eyes had clear cornea after gas tamponade. Retina was reattached under the gas action in 21 eyes during the secondary operation. Visual acuity improved in 22 eyes, remained unchanged in 3 eyes and decreased in 1 eye during the follow-up of 3-12months.· CONCLUSION: Application of pure C3F8 tamponade on or after primary operation can effectively improve hypotony after severe ocular trauma and benefit a better prognosis.

Objective To identify mutations site and clinical characteristics of a familial hypercholesterolemia(FH) proband diagnosed clinically through DNA sequencing and family analysis in the proband and his family members of 3 generations.Methods Blood samples and clinical data of the kindred of total 29 from 3 generations members were collected.Proband had a physical examination electrocar-diogrom and vascular ultrasound.The proband and his family members took routine clinical exams,and genomic DNA was isolated.The promoter region and the 18 exons of low density liporotein receptor(LDLR) gene were screened by Touch down polymerase chain reaction -single strand conformation polymorphism(PCR-SSCP) and DNA sequencing.The result of sequencing were matched gene sequence published in the BLAST database.Results 1.Increased intima-media thickness and plaque were detected in the common carotid artery,right subclavian artery of the proband.Aortic valve regurgitation was found by echocardiography.2.No mutation R3500Q of ApoB100 was observed.3.Two heterozygous mutations in exon 10 and 13 of LDLR gene (W462X and A606T) were identified.The proband and 5 members of paternal relatives showed W462X heterozygosis mutation in exon 10 of LDLR gene which introduced the change from tryptophone to a new stop codon.The proband's mother and grandmother harboured A606T heterozygous mutation in exon 13 of LDLR gene due to a single base pair substitution of G for A in the codon for residue 1 879.Conclusions Disease causing mutations of proband are W462X and A606T compound heterozygosis mutation in exon 10 and 13 of LDLR gene inherited from mother and father.Proband shows homozyous phenotype though the genotype analysis indicates heterozygous mutations.

Objective To determine LDLR gene mutation in 2 clinically diagnosed FH patients from Hubei province and provide basis for gene diagnosis of FH.Methods Clinical data of 2 FH patients and their parents were collected.The promoter region and exon 1 to exon 18 region of LDLR gene were amplified through PCR and the amplified products were analyzed by forward and reverse DNA sequencing.The mutations were identified after comparison with LDLR gene sequence in GenBank.The pathogenic gene mutations were confirmed according to both genotype and phenotype of FH probands.Results The levels of plasma TC of two probands were 12.79 and 11.98 mmol/L.respectively.No gene mutations were detected in region 3 500 to 3 531 of ApoB100. The mutations of LDLR gene were compound heterozygous mutations. The novel mutation 665G > T detected in the exon 4 of No. 1 proband's LDLR gene was heterozygous missense mutation. The novel mutation 1 358 +32C > T was detected in the exon 9 of No. 1 proband's LDLR gene.The mutations 665G > T ( paternal origin) and 1 358 + 32C > T ( maternal origin) were inherited from the parents. A novel mutation 1 257 C > A was detected in the exon 9 of No. 2 proband's LDLR gene, resulting the presence of a premature termination codon, which was different from 1 257 C > G reported in Belgium.Another heterozygous missense mutation 1 879 G > A was detected in exon 13. They were derived from paternal origin and maternal origin, respectively. Conclusions There are three novel gene mutations:665G >T, 1 358 +32C > T, 1 257C > A found in two probands with compound heterozygous mutations in LDLR respectively. They maybe play a potential role in FH pathogensis.

Objective To explore the association between sleep quality and the increasing risk of type 2 diabetes mellitus (T2DM).Methods A total of 771 patients aged 25-70 years living in Xuzhou City of Jiangsu Province for at least 5 years were enrolled for the survey of risk factor related noninfectious chronic disease in 2013.In this investigation,those who suffered from other types of diabetes,neuropathy,other endocrine disease,cardiovascular,renal and hepatic dysfunction,dyspnea or cancer were excluded.To reduce the influence of confounding factors,another 771 participants were enrolled as controls.Each case was arranged to have a control who was matched in age (difference not more than 3 years),gender,residence and family history.All the participants were interviewed with self-designed questionnaire,and sleep quality was measured by Pittsburgh Sleep Quality Index (PSQI) questionnaire.Student's t test,Chi-square and multivariate logistic regression were used for data analysis.Results The PSQI score in the T2DM patients vs.the controls were 5.15±2.40 vs.2.71 ± 1.93 (t=21.96,P＜0.01).The scores of sleep-related factors,including subjective poor sleep quality,bedtime resistance,short sleep duration,sleep efficiency,sleep disturbance,use of sleep medication and daytime dysfunction,of the T2DM patients were higher than those of the controls.The proportion of sleep related behaviors of the T2DM patients was higher,except for early awakening,cold feeling and nightmare.Poor sleep quality was associated with the increasing risk of T2DM (odds ratio 2.06,95％ CI 1.69-2.52).In multivariate logistic regression,when adjusted for confounding factors,the risk of T2DM was still increased (odds ratio 1.72,95％ CI 1.62-1.83).Sleep-related factors (e.g.subjective poor sleep quality,bedtime resistance,short sleep duration,sleep efficiency and sleep disturbance) were correlated with the risk of T2DM (odds ratio was 3.34,1.63,1.10,1.87 and 3.89,respectively).Conclusion Low quality of sleep may be strongly associated with an increased risk of T2DM.

The treatment and signaling pathway regulation effects of kidney- tonifying traditional Chinese medicine on osteoporosis have been widely studied, but without a systematic summary currently. This review comprehensively collected and analyzed the traditional Chinese medicine on the treatment and signaling pathway regulation of osteoporosis in recent ten years, such as Epimedium, Drynariae Rhizoma, Cnidium, Eucommia, Psoralen and Dipsacus. Based on the existing findings, we concluded the following conclusions: (1) kidney-tonifying traditional Chinese medicine treats osteoporosis mainly through BMP-Smads, Wnt/β-catenin, MAPK, PI3K/AKT signaling pathway to promote osteoblast bone formation and through OPG/RANKL/RANK, estrogen, CTSK signaling pathway to inhibit osteo?clasts of bone resorption. ① Epimedium, Drynariae Rhizoma, Cnidium and Psoralen up-regulate the key proteins and genes of BMP-Smads and Wnt/β-catenin signaling pathways to promote bone formation.② Epimedium, Drynariae Rhizoma, Cnidium, Eucommia, Psoralen, Dipsacusinhibit the bone resorption by mediating the OPG/RANKL/RANK signaling pathway. (2) Kidney-tonifying traditional Chinese medicine prevent and treat osteoporosis through a variety of ways: Icariin, Naringin, Osthol, Psoralen can regulate BMP-Smads, Wnt/β-catenin signaling pathway to promote bone formation, but also activate OPG/RANKL/RANK, CTSK and other signaling pathway to inhibit bone resorption. (3) The crosstalk of the signaling pathways and the animal experiments of the traditional Chinese medicine on the prevention and treatment of osteoporosis as well as their multi-target mechanism and comprehensive regulation need further clarification.

Objective To explore the molecular basis of familial hypercholesteraemia(FH)by analyzing the phenotype and genotype relationship through identify the low density liporotein receptor(LDL-r)gene mutation in a FH kindred.Methods A male patient of 15 years old was selected to examine the electrocardiogram,lipid.Color Doppler was used to examine heart and great vessels.The promoter region and the 18 exons of the LDL-r gene were screened by touch-down polymerase chain reaction(PCR)and DNA sequencing.Results The caro-tid intima-media thickness(IMT)was increased to 0.23 cm,while coronary flow velocity reserve(CFVR)was decreased to 1.57,and mode-rate mitral regurgitation was found in the proband.The genetic alteration G→A change at 1 448 of exon 10 causing premature stop codon(W462X).The same heterozygous nonsense mutation was also found in his father.The mutation had been reported in other Chinese patients.In vitro experiments showed that W462X mutation leads to low LDL binding and internalization ability.Conclusions The homozygous mutation(W462X)in exon 10 of the LDL-r gene were identified in the clinically heterozygous FH proband.The W462X mutation is the underl-ying cause of hypercholesterolaemia and clinical AS manifestations.W462X is recurrent mutation among Chinese FH patients.It might be a hot spot mutation in LDL-r in Chinese FH.J Appl Clin Pediatr,2009,24(1):18-20

OBJECTIVETo investigate the role of the non-canonical Wnt signaling pathway in development of non-alcoholic steatohepatitis (NASH) related to type 2 diabetes mellitus (T2DM) using a rat model.

METHODSTwenty-four male Sprague-Dawley rats were randomly divided into two equal groups: control group, fed a stand diet; T2DM-NASH model group, fed a high sucrose and fat diet for 4 weeks and intraperitoneally injected with streptozotocin (30 mg/kg). Twelve weeks after model establishment, all rats were sacrificed. Serum levels of glucose, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected by biochemical analysis. Liver pathological changes were assessed microscopically by hematoxylin-eosin and oil red O staining. The liver expression of Wnt5a and NF-kB p65 were detected by immunohistochemistry and western blotting (protein), and quantitative real-time PCR (mRNA).

RESULTSThe T2DM-NASH model group showed significantly higher levels of glucose (control group: 6.25 +/- 1.28 vs. 31.21 +/- 0.86 mmol/L, t = -36.204, P less than 0.01), ALT (31.00 +/- 3.69 vs. 301.50 +/- 8.62 U/L, t = -99.94, P less than 0.01), and AST (77.58 +/- 1.83 vs. 344.75 +/- 1.82 U/L, t = -358.85, P less than 0.01). The T2DM-NASH group also showed remarkable signs of steatosis and inflammation in hepatic tissues. The T2DM-NASH group had significantly higher integral optical density (IOD) detection of Wnt5a (control group: 1.15E4 +/- 577.45 vs. 4.04E5 +/- 2.42E4, t = -56.24, P less than 0.01) and NF-kB p65 (1.28E4 +/- 1.59E3 vs. 4.21E5 +/- 1.68E4, t = -83.895, P less than 0.01), as well as protein levels detected by western blot (Wnt5a: 4.21 +/- 0.34 vs. 1.00 +/- 0.25, t = 17.030, P less than 0.01; NF-kB p65: 4.93 +/- 0.76 vs. 1 +/- 0.13, t = 11.438, P less than 0.01). The hepatic mRNA levels followed the same trend (Wnt5a: 9.53 +/- 0.64 vs. 1.04 +/- 0.35, t = 20.165, P less than 0.01; NF-kB p65: 0.60 +/- 0.13 vs. 0.74 +/- 0.10, t = -1.802, P = 0.125). In the T2DM-NASH group, hepatic Wnt5a protein expression was positively correlated with ALT (r = 0.64, P less than 0.05), AST (r = 0.59, P less than 0.05), and NF-kB p65 protein expression (r = 0.58, P less than 0.05).

CONCLUSIONWnt5a may activate NF-kB to stimulate an inflammatory response leading to development of NASH related to T2DM.

Animals ; Diabetes Mellitus, Experimental ; complications ; metabolism ; Diabetes Mellitus, Type 2 ; complications ; metabolism ; Liver ; pathology ; Male ; Non-alcoholic Fatty Liver Disease ; etiology ; pathology ; Rats ; Rats, Sprague-Dawley ; Transcription Factor RelA ; metabolism ; Wnt Proteins ; metabolism ; Wnt Signaling Pathway ; Wnt-5a Protein