Objective To investigate the association between the single nucleotide polymorphisms (SNPs) rs894278 and rs11931074 of α-synuclein (SNCA) and non motor symptoms in Parkinson' s disease (PD).Methods One hundred and twenty PD patients and 100 healthy controls enrolled from Tonglu Hospital Affiliated to Ruijin Hospital and Ruijin Hospital,Shanghai Jiaotong University School of Medicine from 2012 to 2014 were recruited and the motor subscale of the Unified Parkinson' s Disease Rating Scale Ⅲ (UPDRS-Ⅲ) was used to evaluate motor function.The Mini-Mental State Examination (MMSE),17-item Hamilton Rating Scale (HAMD-17),the Rapid Eye Movement Behavior Disorder Screening Questionnaire (RBDSQ) and the ROME-Ⅲ criteria for chronic constipation were used to evaluate non motor symptoms.SNCA SNPs (rs894278,rs11931074) were genotyped by direct sequencing.Results There was no statistically significant difference in age,sex,non motor symptoms scores among the three genotypes of the two SNPs in control group.There was no statistically significant difference in age,sex,UPDRS-Ⅲ scores and Hoehn-Yahr stage among the three genotypes of the two SNPs in PD group.The results demonstrated that there was no association between the two SNPs and RBDSQ scores,HAMD-17 scores,MMSE scores and constipation in PD patients.However,additional analysis showed that patients with GG rs894278 had a greater proportion of clinical probable RBD than those with GT and TT types (GG 52.2％,12/23;GT 18.2％,10/55;TT 21.4％,9/42;x2 =9.254,P=0.002;x2 =6.424,P=0.005).In Logistic regression analyses adjusting for age and sex,we observed that rs894278 GG genotype could increase the risk of RBD in PD patients (OR =5.367,95％ CI =1.607-17.925,P =0.006).There was no association of RBD with allelic and genotypic distributions of SNCA rs11931074.Conclusion The results indicate that the rs894278 polymorphism correlates with RBD,while rs11931074 does not.

BACKGROUND: It is difficult to in vitro isolate and culture the ovarian surface epithelium with high purity, strong vitality and stable biological characteristics. Tissue adherence and enzymatic digestion are commonly used for primary culture, but there are certain problems in cel col ection, cel viability and cel purity. OBJECTIVE: To establish a method for primary isolating, culturing and identifying human ovarian epithelium. METHODS: The ovarian surface epithelium was obtained with cel brush innovatively, and then the cells were isolated and purified with erythrocyte spal ation and differential adherence. The epidermal growth factor was added into the serum-free Dulbecco’s modified Eagle’s medium-F12 medium for cel culture. The cel morphology was observed under inverted microscope, and hematoxylin-eosin staining and immunocytochemical staining were used to identify the cells, then the growth curve was draw. RESULTS AND CONCLUSION: The ovarian surface epithelium became adherent after cultured for 24 hours, and reached fusion basical y after cultured for 6-12 days. The cells were polygonal or flat with strong transparency and refraction. The morphological characteristics of the cells were in line with those of the normal epithelial cells, and almost al the isolated cells could express the epithelial cells surface marker CK19. The cells could be passaged for 6-8 generations with wel growth and the cel growth curve was in “S” shape. The purity of the cells was more than 95%. The results suggest that cel brush method is simple to operate and can obtain a large amount of ovarian surface epithelium rapidly. The purity of the isolated cells can reach to 95% after treated with erythrocyte spal ation and differential adherence method and the cells are in stable growth.

Objective To explore the folate receptor mediated (FRD) check and multi-function acetic acid white solution liquid based cervical cytology (TCT) application value in cervical cancer screening.Methods A total of 602 cases of patients was tested with FRD multi-function acetic acid white solution check,and TCT and cervical biopsy pathology examination.With the used of histopathological results as the gold standard,FRD multi-function acetic acid white analysis was compared with the TCT screening inspection results.Results For a total 602 patients with TCT screening,the positive rate was 21.8％ (131/602),including 36 cases of CIN Ⅰ level,41 cases of CIN Ⅱ level,24 cases of CIN Ⅲ level,and 30 cases of cervical invasive carcinoma.For the FRD multifunction white acetate solution screening,its positive rate was 23.8％ (143/602).No statistically significant difference was found between TCT and FRD screening (P ＞ 0.05).The missed diagnosis rate of FRD multi-function white acetate solution screening was 2.6％ in inflammation,and 21.1％ in cervical invasive cancer,and 3.8 ％ in CIN.The missed diagnosis rate of TCT screening was 7.2％ in inflammation,5.3％ in CIN Ⅰ,4.9％ in CIN Ⅱ,and 58.6％ in CIN Ⅲ]; whereas,its detection coincidence rate was 100％ in squamous cells carcinoma (SCC) and adenocarcinoma (AC).FRD multi-function acetic acid white solution screening had a sensitivity 80.92％,specificity 92.14％,positive predictive value 74.13％,and negative predictive value 95.59％.TCT examination had a sensitivity 90.84％,specificity 90.23％,positive predictive value 72.12％,and negative predictive value 97.25％.No significant difference was found between FRD and TCT methods (P ＞ 0.05).Conclusions FRD and TCT methods were both efficient in screening and evaluation for cervical lesions and cervical cancer.Because FRD method is limited in the deep tube for examination of cervical lesions; it cannot completely replace the TCT examination.However,FRD method is reliable,economic,and simple operation; it is suitable for primary hospitals census of cervical cancer

Objective To investigate treatment effects of lentivirus mediated RhoA short hairpin RNA(shRNA) on xenograft tumor of ovarian cancer in nude mice in vivo and the underlying mechanism.Methods Human ovarian cancer cell line HO8910 were inoculated to establish subcutaneous xenograft model of human ovarian cancer.Tumor-bearing nude mice were assigned randomizely to three groups:LentiRhoA-sh group,Lenti-negative control (NC) group and phosphate buffered saline (PBS) group.lentivirus mediated RhoA shRNA,negative control lentivirus and PBS were respectively injected in the three groups.Effects of treatment were observed by tumor growth curve,tumor volume,tumor weight,and tumor inhibition rate.Xenograft tissues and liver,spleen,lung,and renal tissues were examined by hematoxylin and eosin (HE) staining or were detected by streptavidin-perosidase (SP)immunochemical method.The changes of RhoA gene expression in xenograft tissues after lentivirus mediated RhoA shRNA treated were also detected by real-time qPCR,immunochemistry and Western blot assay.Cell apoptosis in xenograft tissues were examined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL) method and apoptotic index (AI) were counted.Results Compared with Lenti-NC group and PBS group,the growth speed of xenograft in Lenti-RhoA-sh group delayed significantly after injection 9 days(P ＜ 0.01).Tumor volume (338 ± 114) mm3 decreased significantly in the Lenti-RhoA-sh group when compared with those in Lenti-NC group(1190 ± 332)mm3 and PBS group (1101 ± 396) mm3 (P ＜ 0.01).Tumor weight (0.23±0.11)g decreased significantly in the Lenti-RhoA-sh group when compared with Lenti-NC group (0.79 ± 0.19)g and PBS group (0.74 ± 0.17)g (P ＜ 0.01).Real-time qPCR result shown that the expression of RhoA mRNA (0.30 ± 0.05) decreased significantly in the Lenti-RhoA-sh group compared with Lenti-NC group (0.95 ±0.06) and PBS group(1.00 ±0.11 ; P ＜0.01).Western blot result showed that the expression level of RhoA protein decreased significantly in the Lenti-RhoA-sh group (0.14 ± 0.06) compared with those in Lenti-NC group(0.78 ± 0.14) and PBS group (0.75 ± 0.13;P ＜ 0.01).TUNEL staining displayed that AI significantly increased in the Lenti-RhoA-sh group (20.9 ± 3.4) ％ compared with those in Lenti-NC group(5.2 ±±2.0)％ and PBS group(6.0 ±2.1)％ (P ＜0.01).Conclusion Lentivirus mediated RhoA shRNA may be effectively down-regulate of the expression of RhoA,inhibit the growth of subcutaneous xenograft tumor of ovarian cancer in nude mice by increasing the cell apoptosis.

Objective To explore the relationship between the expression of transient receptor potential cation channel subfamily C member 6 (TRPC6) and podocyte injury in children with primary nephrotic syndrome (PNS) and its clinical significance. Methods The renal tissue of 18 children with PNS was obtained. The pathological changes of kidney were observed by routine section staining and light microscopy. The structural changes of podocyte were observed by electron microscope. The mRNA and protein expressions of TRPC6 in tissues were determined by qPCR and immunohistochemistry, respectively. Further the correlation of TRPC6 mRNA with serum levels of albumin (Alb), creatinine (Cr), triacylglycerol (TG), cholesterol (Tch), complement C3 and 24 h urinary protein quantitation and estimated glomerular filtration rate (eGFR) were analyzed respectively. Results The expression of TRPC6 protein in renal tissue of children with PNS was higher than that in the control group, and the difference was statistically different (P<0.05). The relative expression of TRPC6 mRNA in renal tissue of children with PNS was positively related to the expression of TRPC6 protein (r=0.508, P<0.05), but there was no correlation of expression of TRPC6 mRNA with serum levels of Alb, Cr, TG, Tch, C3, eGFR and 24h urinary protein quantitation (P>0.05). Conclusion The pathological types of PNS were mainly podocyte lesions, and the expression of TRPC6 protein was increased in podocytes. TRPC6 detection may be helpful in the diagnosis of podocyte lesions.

Objective To investigate the effects ofWenyujinessential oil on tau protein phosphorylation in mice with Alzheimer Disease (AD); To discuss the relevant mechanism of action.MethodsSixty Kunming mice were divided into six groups randomly: sham-operation group, model group,Wenyujin essential oil low- and high-dose group, donepezil group andShenzhiling group.β-Amyloid protein (Aβ25-35) was infused into the hippocampal of mice. Mice in each group were given relevant medicine for gavage for 15 d. After the last administration, immunohistochemistry was used to detect the expressions of tau protein phosphorylation points (Ser 404 and Thr 231). Western blot was used to detect tau protein phosphorylation points and the expression of PI3K/Akt protein. Results Compared with sham-operation group, tau protein phosphorylation points in the model group increased (P<0.05), but the phosphorylation levels of PI3K and Akt decreased significantly (P<0.05). Compared with the model group, tau protein phosphorylation (Thr231 and Ser404) inWenyujin essential oil high-dose group decreased significantly (P<0.05), and the phosphorylation levels of PI3K and Akt increased(P<0.01). ConclusionWenyujinessential oil can inhibit tau protein phosphorylation in mice. The possible mechanism may be related to the PI3K/Akt signal pathway.

Objective To investigate the relationship between apparent diffusion coefficient (ADC)value of rectal adenocarcinoma on DWI and its pathological grading.Methods The ADC values of 46 rectal adenocarcinomas were measured and compared with their histopathological grades.Results The 46 rectal adenocarcinomas included well differentiated adenocarcinomas in 14,moderate-ly differentiated ones in 20,and poorly differentiated ones in 12.The ADC values of well,moderately and poorly differentiated ade-nocarcinomas were (1.125±0.103)×10 -3 mm2/s,(1.030±0.098)×10 -3 mm2/s and (0.922±0.091)×10 -3 mm2/s,respective-ly,exhibiting a statistical difference (χ2 =1 7.35 1,P =0.000).Mann-Whitney U test showed that difference in ADC value between different histopathological grades was statistically significant.Conclusion ADC value of rectal adenocarcinoma can be used as a bio-marker for cell grading to guide treatment decision and prognosis assessment.

ObjectiveTo compare the effects on left ventricular remodeling of bone marrow mononuclear cells transplantation one week and three months after acute myocardial infarction.MethodsAcute myocardial infarction models were successfully established in 15 swine,which were randomly divided into three groups:placebo group,early transplantation group and late trasplantation group.One week after model had been established,early transplantation group underwent bone marrow mononuclear cells transplantation,and then so did the late trasplantation group three months after acute myocardial infarction.B-ultrasound and single photon emission computed tomography (SPECT) examinations were performed to assess the left ventricular end diastolic dimension( LVd),left ventricular end diastolic volume(EDV) and left ventricular ejection fraction(EF) before and one week,three months,six months after acute myocardial infarction.ResultsThe LVd of late transplantation group [ (54.20:t:3.70) mm] was lower than placebo group [ (63.20 ±5.63)mm],but higher than early transplantation group [ (47.40 ± 1.14)mm].The EDV of late transplantation group [ ( 163.00 ±6.96)ml] was lower than placebo group [ (209.40 ± 18.69)ml],but higher than early transplantation group [ ( 135.40 ± 4.93 ) ml ].The EF of late transplantation group (0.25 ±0.02) was higher than placebo group (0.19 ±0.02),but lower than early transplantation group (0.37 ±0.02).ConclusionsBone marrow mononuclear cells transplantation can alleviate progressing ventricular remodeling,even it is performed three month after acute myocardial infarction,but its therapeutic effects are inferior to early transplantation's.

OBJECTIVETo identify potential mutation of the PAX6 gene in a family affected with congenital aniridia from northeastern China.

METHODSTwo patients were collected from the family and underwent full ophthalmologic examinations. Genomic DNA was extracted from all family numbers and 100 healthy controls. The coding regions and flanking sequence of the PAX6 gene were amplified by PCR amplification and subjected to bidirectional DNA sequencing.

RESULTSA nonsense mutation (c.718 C>T) was identified in exon 9 in both patients but not in other unaffected families or the 100 healthy controls. However, obvious difference was noted in the phenotype between the two patients. One of the patient has presented irregular cornea, which was infrequently reported.

CONCLUSIONA c.718C>T transitional mutation has been found to underlie the aniridia, which showed an autosomal dominant inheritance pattern in this northeastern Chinese family.

Aniridia ; genetics ; Eye Proteins ; genetics ; Female ; Homeodomain Proteins ; genetics ; Humans ; Male ; Mutation ; PAX6 Transcription Factor ; Paired Box Transcription Factors ; genetics ; Repressor Proteins ; genetics

Peripherally inserted central venous catheter (PICC)-related upper extremity venous thrombosis (UEVT) is defined as upper extremity venous thrombosis within the veins where PICCs were placed or adjacent to and may result in pulmonary embolism. Malignancies, previous history of venous thrombosis and malposition are common risk factors for PICC-UEVT. Once patients demonstrate clinical manifestations of phlebitis and thrombosis, such as swelling, pain and tenderness at the PICC site, venous duplex ultrasonography is the first choice for diagnosing PICC-UEVT. According to American College of Chest Physicians guidelines, it's not recommended to remove PICCs upon detection of PICC-UEVT. The first-line treatment is to administer systemic anticoagulants while keeping the catheter in place, unless any contraindications. PICCs could continue to be used during anticoagulation therapy, suppose that catheter tip remains well placed and functions as normal. With early diagnosis and standard anticoagulant treatment, a better clinical outcome could be achieved. Prophylactic anticoagulation is not routinely recommended per guidelines. Recommendation for asymptomatic PICC-related thrombosis is still absent and warrants further prospective studies with large sample size.