To study the link between BRAFV600E status and the expression of BIM gene in papillary thyroid carcinoma( PTC) tissues and to analyze the association of these factors with clinicopathological characteristics. BRAFV600E status was determined by MASA-PCR, and qPCR was applied to detect the expression of BIM gene. Finally, the associations of these factors with clinicopathological characteristics were analysed. The rate of mutant BRAFV600E in PTC was 54. 1% , and the expression of BIM gene was lowered in BRAFV600E positive PTC tissues. Additionally, there was significant association( P < 0. 05) between BRAFV600E positiveness and raised TNM Staging (Ⅲ/ Ⅳ), and lowered BIM expression was significantly associated (P<0. 05) with the tumor size and raised TNM Staging(Ⅲ/ Ⅳ). These findings may help us to know more about the mechanism of PTC and to develop new diagnostic biomarkers or prognostic indicators of PTC.

Objective To compare the clinical and pathological features of Chinese young breast cancer(age ≤ 35)with elder patients(> 35)using Meta analysis .Methods Published studies concerning clinical and pathological features of young breast cancer in China were searched systemically and assessed .Stata12 .0 software was used for data analyzing and calculating OR and its 95%CI .Results Totally 31 studies were selected for Meta analysis ,and most of them were classified as 6 - 7 scores ,which showed the quality of articles was high .The risk factors of breast cancer and its pooled odds ratio values with statistical significance were as fol‐lows 6 .42(95% CI :4 .22 - 9 .79) ,0 .61(95% CI :0 .50 - 0 .74)when clinical staging of 0 - Ⅱ phase or Ⅰ - Ⅱ phase ,2 .25(95% CI :1 .69 - 2 .99)when histological type of Invasive carcinoma ,1 .73(95% CI :1 .23 - 2 .43)when histological grade of III grade ,1 .80 (95% CI :1 .23 - 2 .43)when positive of lymph node metastasis .Conclusion Compared with elder breast cancer ,the clinical and pathological characteristics of young breast cancer were mainly for the high misdiagnosis rate ,the late clinical stage ,the high pro‐portion invasive carcinoma ,the poor histological differentiation and the lymph node metastasising easily ,the hint of young breast cancer screening and treatment may be different principles and measures should be adopted .

Objective To isolate and culture the rat limbal stem cells ( LSCs) in vitro,and investigate briefly their capacity for transdif-ferentiation into neural stem cells ( NSCs) with cytokine EGF,bFGF and RA.Methods LSCs derived from rats were cultured and identified by immunohistochemistry in vitro.LSCs were induced to differentiate into NSCs in the presence of EGF (20 ng/mL) ,bFGF (10 ng/mL) and with or without of RA(group 1 or group 2)(25 ng/mL) for seven days.Cultures without factors were used as control group.Then the neural marker Nestin of the coultured cells were measured by immunohistology staining.Furthermore,the positive cell rate was counted under micro-scope between the 2 groups and analyzed by statistical software.Results It showed that P63 was positive in LSCs.Nestin in both of the dif-ferentiation groups was positive at the rate of (77.01 ±6.32)%and (84.01 ±5.43)%respectively,of which the second group was higher than the first one (P<0.05).However,it was negative in the control group.A band of Nestin protein from cells was detected by western blot assay.Conclusion LSCs are successfully isolated and cultured in vitro.LSCs could be induced to differentiate into NSCs in the presence of EGF and bFGF.Moreover,the differentiation capability is enhanced in the condition of RA.

Objective To explore the roles of autoantibodies against ?1 adrenoceptor(?1-receptor)and M2 cholinergic receptor(M2-receptor)in patients with chronic renal insufficiency.Methods The epitopes of the second extracellular loop of ?1 receptor and M2 receptor were synthesized and used respectively to detect the sera autoantibodies against ?1 receptor and M2 receptor by enzyme linked immunosorbent assay(ELISA) in 76 patients with chronic renal insufficiency,60 cases with hypertension and 40 healthy controls.Results In patients with chronic renal insufficiency,the positive rates of the autoantibodies against ?1-receptor and M2-receptor were 56.7% and 38.1% respectively,which were much higher than those of patients with hypertension(18.3% and 11.7%) and higher than those of healthy controls(17.5% and 15.0%)(all P

Objective To explore the role of the autoantibodies against ?_1-adrenergic receptor(?_1-receptor)in the development of hypertension with renal failure.Methods The epitopes of the second extracellular loop of ?_1-receptor were synthesized and used respectively to screen sera autoantibodies from patients with hypertension with renal failure(61 cases),hypertension without renal failure(58 cases) and healthy blood donors(40 cases,control) by ELISA method.Results The positive rates of the autoantibodies ?_1-receptor(69%,42/61) in patients with hypertension with renal failure were higher than those of patients with hypertension without renal failure(19%,11/58) respectively(P

Objective: To analyze the basic thyroid hormone level on long term prognosis and peri-operative recovery in patients after heart transplantation (HT) at the terminal stage of heart failure (HF).
Methods: A total of 270 consecutive patients who received HT at the terminal stage of HF in our hospital from 2009-09 to 2014-07 were retrospectively studied. According to serum levels of thyroid stimulating hormone (TSH), the patients were divided into 3 groups: TSH < 0.55 mIU/L group, TSH (0.55-2.5) mIU/L group and TSH > 2.5 mIU/L group. The peri-operative recovery condition and long term prognosis were observed and compared among 3 groups.
Result: The average age of patients was at (44.58 ± 13.30) years including 228 (84.4%) male and 42 (15.6%) female. The average post-operative follow-up period was (31.88 ± 17.48) months with 100% follow-up rate. There were 41.8% of patients with hypothyroidism, and 46 (17.0%) patients with low T3 syndrome, 56 (20.7%) with subclinical hypothyroidism and 11 (4.1%) with clinical hypothyroidism. The ratio of low level thyroid hormone in HT patients was much higher than general population. For peri-operative recovery, the ICU stay time and mechanical ventilation time were similar among 3 groups,P>0.05, while TSH (0.55-2.50) mIU/L group had the shortest times and TSH > 2.50 mIU/L group had longest times. For long term prognosis, no matter uni-/multi- aviate regression analysis or Kaplan-Meier surviving curve all suggested that TSH > 2.50 mIU/L was the independent risk factor inlfuencing the prognosis of HT patients at the terminal stage of HF. Upon TSH increasing, the patients would have worse prognosis accordingly.
Conclusion: Serum level of TSH > 2.50 mIU/L was the independent risk factor in HT patients at the terminal stage of HF.

To observe the relationship between positive rate of β1-adrenergic and AT1 receptors autoantibodies with serum concentration of cystatin C in 371 patients with diabetic nephropathy patients,107 patients with type 2 diabetes,and 47 subjects as healthy control.In patients with diabetic nephropathy,the positive rates of the β1 and AT1 receptors autoantibodies were significantly higher than those in patients with type 2 diabetes and normal controls.The titers of β1 and AT1 receptors autoantibodies in diabetic nephropathy patients with abnormal cystatin C were significantly higher than those with normal cystatin C concentration.These findings suggested that β1 and AT1 receptors autoantibodie may play important roles in the pathogenesis of diabetic nephropathy.

OBJECTIVETo investigate the optimal starvation conditions of human umbilical vein endothelial cells (HUVECs) and establish a highly efficient and stable method for separating HUVECs.

METHODSHUVECs harvested from human umbilical cords by digestion with 0.1% collagenase II for 15 min were cultured in endothelial culture medium (ECM) containing 5% fetal bovine serum (FBS), 1% endothelial cell growth factor (ECGS) and 1% penicillin/streptomycin solution(P/S) at 37 degrees celsius; in 5% CO2. The cells were observed for cell morphology under an inverted microscope and identified with immunofluorescence assay. The purity of HUVECs was detected using flow cytometry (FCM). The cell cycles of HUVECs cultured in the presence of 0, 0.1%, 0.5%, and 1% FBS for 0, 6, 12, 18, and 24 h were analyzed with flow cytometry.

RESULTSs The purity of HUVECs harvested by digestion with 0.1% collagenase II reached 99.67%. The primary HUVECs showed a cobblestone or volute appearance in vitro. Immunocytochemistry showed that HUVECs highly expressed VIII-related antigen. Cell culture in the presence of different concentrations of FBS for 6 h resulted in 70% G0/G1 phase cells, which increased to 80%-90% at 12 h of cell culture, and further to around 95% at 18 and 24 h.

CONCLUSIONDigestion with 0.1% collagenase II can obtain high-purity primary HUVECs. Culturing HUVECs in serum-free medium for 12 h can result in a high purity (over 80%) of G0/G1 phase cells.

Cell Culture Techniques ; Cell Cycle ; Cells, Cultured ; Culture Media ; chemistry ; Flow Cytometry ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Matrix Metalloproteinase 8 ; chemistry ; Serum

Objective To detect the positive rates of antibodies against avian influenza virus (AIV) subtypes H5, H6, H7 and H9 among people in poultry occupations in Guangdong province and to analyze the transmission of various subtypes of AIV from poultry to human contacts for the prevention and control of novel AIV infection in human beings.Methods Serum specimens were collected from 1066 peo-ple in poultry occupations ( occupational group) and 205 people not in poultry occupations ( non-occupational group) in 10 cities of Guangdong province.The inactivated AIV strains, isolated from poultry or environment of Guangdong province, were used as antigens to detect antibodies against AIV subtypes H5, H6, H7 and H9 by using the hemagglutination inhibition ( HI) assay.Results The positive rates of antibodies against AIV subtypes H5, H6, H7 and H9 carried by people from the occupational group were respectively 0.44%, 0%, 0.30%and 0.30%in 2013 and 1.08%, 0.0%, 0.0%and 0.27%in 2014.Only the anti-H9 anti-bodies were detected in serum samples collected form people in the non-occupational group in 2013 with a positive rate of 0.95%.No significant differences with the positive rates of anti-AIV antibodies were found between the occupational group and the non-occupational group.However, the geometric mean titer ( GMT) of anti-AVI antibodies in people from the occupational group was higher than that of the non-occupational group.Conclusion Although a grand spread of AIV from avian to human is not likely to happen yet, con-tacting with poultry is the risk factor for AIV infection in Guangdong population.A long-term surveillance of anti-AIV antibodies in serum should be strengthened among people in poultry occupations for the timely pre-vention and control of novel AIV outbreak.

OBJECTIVETo study the value of combined use of paternally imprinted gene product p57(KIP2) immunohistochemistry and flow cytometry in the differential diagnosis of placental hydropic diseases.

METHODSA total of 32 cases of hydropic placenta with DNA polymorphism information were collected, and the genetic results were used as basis for the diagnosis of complete hydatidiform moles (CHM), partial hydatidiform moles (PHM) or hydropic abortions. All cases were examined by histology, p57(KIP2) immunohistochemical staining (EnVision method) and flow cytometry DNA ploidy analysis. The p57(KIP2) immunohistochemical staining and DNA ploidy results were compared with the genetic results.

RESULTSIn CHM, p57(KIP2) negative rates were 95.2% (20/21), whereas all the 11 cases of non-CHM (7 cases PHM and 4 cases hydropic abortions) were positive (11/11). In 11 p57(KIP2) -positive cases, 7 cases with triploidy and 4 cases with diploidy by flow cytometry were proven to be PHM and hydropic abortions by genetic analysis, respectively. Overall, 96.9% (31/32) cases of hydropic placentas were correctly diagnosed by combined use of p57(KIP2) immunohistochemistry and flow cytometry.

CONCLUSIONSp57(KIP2) immunohistochemical negativity is a reliable index for the diagnosis of CHM. Combined flow cytometry DNA ploidy and p57(KIP2) immunohistochemistry are useful in the pathological differentiation of CHM, PHM and hydropic abortions.

Abortion, Spontaneous ; diagnosis ; genetics ; metabolism ; Adult ; Cyclin-Dependent Kinase Inhibitor p57 ; metabolism ; DNA, Neoplasm ; analysis ; Diagnosis, Differential ; Diploidy ; Female ; Flow Cytometry ; Humans ; Hydatidiform Mole ; diagnosis ; genetics ; metabolism ; Immunohistochemistry ; Middle Aged ; Pregnancy ; Triploidy ; Uterine Neoplasms ; diagnosis ; genetics ; metabolism ; Young Adult