Objective To evaluate the effects of the different treatments for vitiligo in the special sites. Methods 122 cases (375 leuk oplakiae) of vitiligo in special sites were randomly divided into 5 groups:hair germ grafting (HGG) group (42 cases), resurfacing (RS) group (52 cases), follicular scraping injection (FSI) group (35 cases), liquid nitrogen freezing (LNF) group (30 cases) and external medication (EM) group (45 cases). Efficacy of the treatments was observed and evaluated in different groups and all the data were statistically analyzed. Results Effective rates were 97.1 % in HGG group, 94.7 % in RS group, 59.7 % in FSI group, 57.1 % in LNF group and 45.6 % in EM group.There were very significant differences between different groups ( ? 2 = 111.7, P

Objective To observe the effects of Yougui Pills (YP) intervention on SREBP pathway related factors of sterol regulatory element binding protein-1c ( SREBP-1c) , cholesteryl ester transfer protein ( CETP) and fatty acid synthase ( FAS) protein and gene expression in kidney yang deficiency hyperlipidemia rat model. Methods Sixty SD rats were divided into normal control group, model group (in the dosage of 2.43 g/kg), and YP group. Rat model of kidney yang deficiency hyperlipidemia was induced with intramuscular injection of hydrocortisone. Normal control group and model group were given normal saline, and YP group was given YP suspension intragastrically. After treatment, the hepatic pathomorphology of rats in the three groups was examined, and the contents of serum lipids were examined. Hepatic SREBP-1c, CETP and FAS protein expression was detected by Western-blotting method, and their mRNA expression was detected by real-time polymerase chain reaction ( RT-PCR) . Results Compared with the normal control group, serum triglyceride ( TG ) , total cholesterol ( TC ) , low-density lipoprotein cholesterol ( LDL-C ) contents were significantly increased (P<0.01), high-density lipoprotein cholesterol (HDL-C) was significantly decreased (P<0.01), and hepatic pathomorphological changes of hyperlipoidemia were obvious in the model group. Compared with the model group, TG, TC, LDL-C contents were significantly decreased ( P<0.01) , and HDL-C was significantly increased (P<0.01) in YP group. Model group had higher SREBP-1c, CETP and FAS mRNA and protein expression than the normal control group ( P<0.05) , while YP group had lower SREBP-1c, CETP and FAS mRNA and protein expression than the model group ( P<0.05) . Conclusion YP can decrease the blood lipid levels probably by down-regulating the expression levels of SREBP-1c, CETP and FAS gene and protein related to the the SREBP pathway in rats of kidney yang deficiency hyperlipidemia induced by intramuscular injection of hydrocortisone.

Periampullary carcinoma is a rare malignant tumor of digestive system,and its accurate diagnosis is still difficult.From January 2007 to July 2012,12 patients with periampullary carcinoma had been admitted to the Southwest Hospital of Third Military Medical University,and the imaging data were retrospectively analyzed.The results of ultrasonography revealed that all tumors were hypoechoic.The tumor displayed hyperenhancement in 3 patients,isoenhancement in 1 patient,hypoenhancement in 8 patients during the arterial phase on contrastenhanced ultrasonography (CEUS),while the tumor displayed hypoenhancement in all patients during the venous phase.Magnetic resonance imaging (MRI) plain scanning showed duodenal papilla enlargement in 1 patient,ampullary tissue mass signal in 2 patients,tissue mass signal at distal common bile duct in 2 patients,the rest 7 patients did not show tissue mass signal.Lower biliary obstruction was the common manifestation of the 12 patients on magnetic resonance cholangiopancreatography (MRCP),intrahepatic and extrahepatic bile vine-like expansion in 4 patients,double duct sign in 2 patients,the bottom of common bile duct with filling defect in 2 patients and it revealed beak-like narrow in 1 patient.CEUS,MRI and MRCP could both play an important role as conventional methods in diagnosing periampullary carcinoma.

AIM: To observe the effect of E2F decoy DNA on proliferation and apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS: E2F decoy DNA,ARE decoy DNA and control decoy DNA were transfected into PC-3M cells with lipofectamine,respectively.Their effects on cell proliferation were detected by MTT assay.The changes of cell morphology were observed by inverted phase contrast microscope.The cell apoptotic rate was determined by flow cytometry(FCM) analysis and chromosome DNA ladder was detected by DNA gel electrophoresis.The expression of c-Myc mRNA and cyclin D1 mRNA was detected by RT-PCR.The protein levels of c-Myc and cyclin D1 were detected by Western blotting.RESULTS: The growth of PC-3M cells was inhibited after transfection.The transfected PC-3M cells displayed typical apoptotic morphological changes.The apoptotic rate was 26.35% and DNA ladder was observed after transfection.The expression of c-Myc and cyclin D1 were inhibited.CONCLUSION: These results indicate that E2F decoy DNA induces apoptosis of androgen-independent prostate cancer cell lines PC-3M and inhibits cell proliferation via inhibiting expression of c-Myc and cyclin D1.

ObjectiveTo detect mRNA and protein expression of steroidogenic factor-1 ( SF-1 ) and DAX-1 in human adrenocortical tumors and normal adrenal cortex,and to investigate the effect of SF-1 and DAX-1 on the steroidogenesis and development of adrenocortical tumors.Methods Total RNA and protein was extracted from angiotensin Ⅱ unresponsive aldoterone-producing adenomas ( A Ⅱ -U-APA,n =12 ),angiotensin Ⅱ responsive aldoterone-producing adenomas ( AⅡ -R-APA,n =5 ),cortisol-producing adenomas ( CPA,n =10 ),adrenal nonfunctional adenomas ( NFA,n =10 ),aldosterone-producing carcinoma ( APC,n =2 ) and normal adrenal cortex ( NAC,n =8).To analyze gene expression of SF-1,DAX-1,ACTH receptor(ACTHR),and β-actin by real-time quantitative PCR in different tissues.The protein expression of SF-1,DAX-1,and β-actin in the same tissues by Western blot.To study the relationship of ACTHR,SF-1,and DAX-1 with clinical data in adrenocortical tumors.ResultsThe expression of SF-1,DAX-1 mRNA and protein was different in NAC,AⅡ -U-APA,A Ⅱ -R- APA,APC,CPA,and NFA tissues [ relative expression of SF-1 mRNA:24.58±2.45,23.89±3.17,21.59±3.00,(38.75,44.16),14.17±2.80,and 36.38±3.50; DAX-1 mRNA:0.57±0.06,0.37±0.05,0.43±0.05,( 1.52,1.21 ),0.39 ±0.04,and 0.83 ±0.08 ; SF-1 protein:0.76 ±0.11,0.76 ±0.10,0.73 ±0.07,(1.24,1.40),0.55±0.04,and0.98±0.10; DAX-1 protein:0.65±0.14,0.39±0.13,0.43±0.14,(1.18,1.02),0.56±0.04,and 1.03±0.13 ; all P＜0.05 or P＜0.01 ].There was negative correlation by higher SF-1/DAX-1 ratio and tumor size in AⅡ -U-APA tissues.The mRNA and protein expression of SF-1 was lower in CPA and there was the positive correlation with tumor size.Conclusion SF-1 and DAX-1 might play a key role in the development of the adrenocortical tumorigenesis and steroidogenic tissues.

AIM: To study the basic mechanism of transcriptional regulation, NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested. METHODS: 1.04 kb - promoter fragment of NKX3. 1 gene was obtained by PCR and cloned into pGL3 - basic and pEGFP - 1 that are promoter - less reporter vectors to examine its promoter activity driving the reporter gene transcription. The promoter activity was determined by dual -luciferase reporter assay and the expression of GFP reporter observed under fluorescence micro scope. RESULTS: The sequence of the cloned 1.04 kb promoter proved to be correct by DNA sequencing. The dual - lu ciferase reporter assay (M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL3 - 1.04 kb promoter was about 1.5 - fold higher than that of pGL3 - control transfection and 50 - fold higher than that of pGL3 - basic transfec tion. To investigate the 1.04 kb - promoter activity in different tumor cell lines, the constructed pGL3 - 1.04 kb promoter and pEGFP - 1.04 kb promoter were transfected into several cell lines, respectively. The results showed that the activity of 1.04 kb promoterin LNCaP was highest among the tested cell lines. Multiple consensus sequence elements have been iden tified within the 1.04 kb fragment using TRANSFAC database. Further experiments will be done to determine their founc tions. CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

AIM: To observe the effect of exogenous androgen responsive element decoy on the promoter of prostate specific antigen (PSA) and the growth of LNCaP cells for searching the possibility of gene therapy for prostate cancer. METHODS: Firstly, pGL3 - PSA luciferase expression vector containing 640bp - promoter fragment of PSA gene was constructed. Then, a 23 -mer phosphorothioated ARE decoy based on the deduced ARE sequence at the promoter region of PSA gene was synthesized. pGL3 - PSA and ARE decoy DNA were cotransfected into PC3 - M cell by lipofectamineTM 2000. Through detecting the activity of luciferase, the effect of ARE decoy on the promoter of PSA was studied. Then the ARE decoy DNA was transfected into LNCaP cells. The effect of decoy DNA on the proliferation of LNCaP cells was examined by using MTT assay. The effects of apoptosis were detected by phase contrast microscopy, DNA agrose gel electrophoresis and flow cytometry. Meanwhile, the nuclear extract was prepared from LNCaP cells and DNA - protein interactions were examined by electrophoretic mobility shift assay (EMSA). RESULTS: The reporter assay showed that the aetivity of luciferase was significantly reduced in the ARE decoy - transfected cells, bnt not in the cells transfected with the control decoy. EMSA demonstrated specific binding of the ARE decoy to androgen receptor. The growth of LNCaP was remarkably inhibited and apoptotic morphological changes as well as DNA fragmentation were observed in the ARE decoy- transfected cells. The rate of apoptosis was 22.4% detected by FCM. CONCLUSION: The ARE decoy is capable of inhibiting the promoter of PSA gene and inducing the apoptosis in prostate cancer cells. It may become a potential therapeutic tool for prostate cancers.

Purpose To investigate the expression and clinical significance of Foxp3 ( cell surface marker of regulatroy T) mRNA and its protein in endometrial carcinomas and normal endometrial tissues. Methods Real-time fluorescence quantitative PCR and immuno-histochemical SP methods were used to detect the expressions of mRNA and protein in tumor tissue of 84 cases of endometrial carcino-mas and 40 cases of normal endometrial tissue, then to analyze the relationship between Foxp3 gene and clinical pathological character-istics of endometrial carcinoma specimens, such as differentiation, FIGO stage. Results Foxp3 mRNA and it′s protein expression of endometrial carcinomas were significantly higher than that of normal endometrial tissues. There were significantly relationships between Foxp3 mRNA expression and FIGO stage of endometrial cancer, Foxp3 mRNA expressions of III+IV stage was higher than that ofⅠ+Ⅱ stage endometrial carcinoma (P<0. 05). But the relationship between Foxp3 expression and differentiation degree reached differ-ent conclusions in the two detection methods. By immunohistochemistry the expression of Foxp3 protein was correlation with histological differentiation grade (rs =0. 72, P <0. 01). In poorly differentiated endometrial carcinoma Foxp3 + cell number was significantly higher than that in well differentiated endometrial carcinoma. By detection of real-time fluorescence quantitative PCR method, Foxp3 mRNA expression was not correlated with tumor grade (rs =0. 01, P=0. 35). Conclusion Foxp3 in endometrial carcinomas are high expressions. Immunohistochemical method has more clinical value than real-time fluorescence quantitative PCR test results. Foxp3 may be involved in the regulation of the development of endometrial cancers.

AIM: To study the basic mechanism of transcriptional regulation,NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested.METHODS:(1.04 kb)-promoter fragment of NKX3.1 gene was obtained by PCR and cloned into pGL_3-basic and pEGFP-1 that are promoter-less reporter vectors to examine its promoter activity driving the reporter gene transcription.The promoter activity was determined by dual-luciferase reporter assay and the expression of GFP reporter observed under fluorescence microscope.RESULTS: The sequence of the cloned(1.04 kb) promoter proved to be correct by DNA sequencing.The dual-luciferase reporter assay(M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL_3-(1.04 kb) promoter was about 1.5-fold higher than that of pGL_3-control transfection and 50-fold higher than that of pGL_3-basic transfection.To investigate the 1.04 kb-promoter activity in different tumor cell lines,the constructed pGL_3-(1.04 kb) promoter and pEGFP-1.04 kb promoter were transfected into several cell lines,respectively.The results showed that the activity of(1.04 kb) promoter in LNCaP was highest among the tested cell lines.Multiple consensus sequence elements have been identified within the(1.04 kb) fragment using TRANSFAC database.Further experiments will be done to determine their founctions.CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

Objective To evaluate the value of captopril challenge test (CCT) in the diagnosis of primary aldosteronism (PA).Methods A total of 674 patients [(45.0±13.7) years, men 341, women 333] admitted to Peking Union Medical College Hospital from 2000 to 2015 were analyzed.Among them, 222 subjects were with essential hypertension (EH), 28 were with pheochromocytoma (PHEO), 246 were with idiopathic hyperaldosteronism (IHA) and 178 were with aldosterone producing adenoma (APA).All patients received CCT.24 h urine sodium was measured in partial patients.Plasma renin activity (PRA), aldosterone (ALD) were detected.Results Compared with EH [PRA: before 0.5(0.2,0.9) μg·L-1·h-1, after 0.8(0.4,1.5) μg·L-1·h-1;ALD: before (393±122) pmol/L, after (360±97) pmol/L] and PHEO [PRA: before 0.3(0.1,0.9) μg·L-1·h-1, after 0.4(0.1,1.6) μg·L-1·h-1;ALD: before (396±108) pmol/L, after (374±114) pmol/L], lower levels of PRA and higher levels of ALD before and after CCT were observed in PA patients [PRA: before 0.1 (0.1,0.2) μg·L-1·h-1, after 0.1 (0.1,0.2) μg·L-1·h-1;ALD: before (468±216) pmol/L;after (457±199) pmol/L].After CCT, the suppression rate of ALD [2.8% (-8.8%,15.4%) vs 6.6% (-4.3%, 17.6%)] and increasing rate of PRA [0(0,50%) vs 50%(0, 200%)] in PA patients were lower than those in EH patients.The ALD/PRA ratio (ARR) were higher in PA than that in EH or PHEO patients.In the EH subjects, ALD levels of seated posture were higher than those of recumbent posture both before and after receiving captopril, but with no changes in ARR after CCT.No significant differences in ALD and ARR (before and after receiving captopril) were observed between seated and recumbent position in the PA group.The ARR after CCT tended to decrease in EH subjects with elevated urine-sodium compared with those with normal urine-sodium.No changes could be viewed in ALD and PRA levels between normal urine-sodium and elevated urine-sodium groups among APA, IHA and EH patients either before or after CCT.Among patients with APA, the ALD levels before CCT and the ARR after CCT were lower in the patients with AngiotensionⅡ(AngⅡ) reactive than those without.A ROC curve analysis suggested that the optimal cutoff value was 46.2 (ALD unit:ng/dl;PRA unit:μg·L-1·h-1) for ARR after challenge in diagnosing PA, with the sensitivity of 88.7% and specificity of 84.8%.Conclusions ARR after 25 mg captopril had high sensitivity and specificity in diagnosis of PA with the cutoff of 46.2.Seated CCT could replace recumbent CCT as a more confirmatory test.The PRA increasing rate should be taken into consideration when diagnosis of PA.