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 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 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 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 .

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

BACKGROUND AND OBJECTIVEStudies showed that osteopontin (OPN) regulates cell migration and invasion in a variety of cancers, which associates with the activities of matrix metalloproteinase (MMP)-2 and MMP-9. This study was to investigate the role of OPN in the proliferation and invasion of human prostate cancer PC-3 cells and the possible functions of IgammaB kinase (IKK) in nuclear factor kappa B (NF-kappaB)-mediated signaling pathways.

METHODSOPN short-hairpin RNA (shRNA) recombinant plasmids were transfected into PC-3 cells and different concentrations of IKK inhibitors were used to inhibit the activities of IKKalpha and IKKbeta. The mRNA and protein expression levers of OPN, MMP-2, and MMP-9 were detected by real-time polymerase chain reaction (PCR) and Western blot. Cell cycle was detected by flow cytometry, cell proliferation by MTT assay, and cell invasion by Transwell chamber assay.

RESULTSCompared with untreated cells, the protein levers of OPN, MMP-2, and MMP-9 in OPN shRNA-transfected PC-3 cells were reduced by 55.22%, 51.71%, and 28.35%, respectively, and the ability of cell migration and invasion were decreased by 45.48% and 51.96%, respectively (P<0.05). Moreover, the inhibition of IKKbeta inhibited the expressions of MMP-2 and MMP-9.

CONCLUSIONA shRNA expression vector-mediated OPN gene silencing can inhibit the malignant biological behaviors of PC-3 cells. IKKbeta may play a crucial role in the OPN-induced activation of MMP-2 and MMP-9 via NF-kappaB-mediated IkappaB/IKKbeta pathways.

Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Genetic Vectors ; Humans ; I-kappa B Kinase ; metabolism ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; NF-kappa B ; metabolism ; Neoplasm Invasiveness ; Osteopontin ; genetics ; metabolism ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; Signal Transduction ; Transfection

OBJECTIVETo study the effects of artemether and dihydroarteannuin on the mouse model of scleroderma.

METHODSixty mice were randomly divided into 8 groups: PBS control group, model group, menstruum group (20% Tween-80, 0.4%CMC-Na), positive medicine group (penicillamine 200 mg kg(-1)), low-dose artemether group (5 mg kg(-1)), high-dose artemether group (20 mg kg(-1)), low-dose dihydroarteannuin group (5 mg kg(-1)), high-dose dihydroarteannuin group (25 mg kg(-1)). We have established a mouse model for scleroderma in Balb/c mice by subcutaneous injections of bleomycin 0.1 mL per day (200 mg L(-1) BLM) for 3 weeks. Meanwhile, the administration lasted for 4 weeks. The back skin was removed in the next day after the final administration. Treated skins and lungs were harvested and analyzed for histological sclerosis. The thickness of the skin and fibrosis degree of derma were observed and made an analysis of the contents of collagen and hydroxyproline.

RESULTCompared with the model groups, the high-dose groups markedly inhibited the thickness of derma (P<0.001), furthermore, the contents of collagen and hydroxyproline in the skin were also significantly reduced (P<0.05). Other groups of mice showed improvement on scleroderma.

CONCLUSIONOur results suggest that administration of artemether or dihydroarteannuin may be an effective approach in preventing systemic sclerosis.

Animals ; Artemisinins ; pharmacology ; Collagen ; metabolism ; Female ; Hydroxyproline ; metabolism ; Lung ; drug effects ; pathology ; Mice ; Mice, Inbred BALB C ; Scleroderma, Systemic ; metabolism ; pathology ; prevention & control ; Sesquiterpenes ; pharmacology ; Skin ; drug effects ; metabolism ; pathology