Breast Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; methods ; standards ; Liver Neoplasms ; metabolism ; pathology ; Quality Control ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Reproducibility of Results

Aim To study the effect of oxymatrine (OMT)on protecting human umbilical vein endothelial cells (HUVECs)from toxicity induced by high glucose in vitro.Methods The HUVECs were cultured with medium containing different concentrations of glucose or OMT. The cells were randomly divided into 6 groups:5.5 mmol·L -1 control group(Control),22.2 mmol·L -1 high glucose (22.2 mmol·L -1 ),44.4 mmol·L -1 high glucose (44.4 mmol·L -1 ),control+OMT(control +OMT),22.2 mmol·L -1 high glu-cose +OMT(22.2 mmol·L -1 +OMT),44.4 mmol ·L -1 high glucose +OMT (44.4 mmol · L -1 +OMT).The protective effect of oxymatrine was as-sessed by MTS assays.The expression of A2B in HU-VECs was detected by Real-time quantitative PCR (qPCR)and Western Blot methods.Results The glucose was shown to have caused cytotoxicity in HU-VECs.Oxymatrine (3 μmol·L -1 )was found to have protected HUVECs from glucose toxicity effectively, and reduced the expression of A2B significantly.Con-clusion Oxymatrine can obviously protect HUVECs from cytotoxicity induced by high glucose and the effect is performed partly by decreasing A2B expression.

Objective To investigate the feasibility and clinical application of prospective ECG-gated scan and adaptive cardio se-quence scan of dual-source CT coronary angiography.Methods 255 patients who underwent coronary CT angiography using a dual-source CT scanner were divided into two groups:group A underwent prospective scan,group B underwent retrospective scan.The two groups were further subdivided into low heart rate group and high heart rate group,recorded as A1(n=51),A2(n=89),B1(n=26), B2(n=89),respectively.Group A1 preceded with prospective ECG gated scan;group A2 performed with adaptive cardio sequence scan;group B1 and B2 underwent retrospective scan.All of them applied ATCM (automatic tube current modulation)technique. Effective dose was recorded.The image quality was assessed by two experienced radiologists with a four-point grading scale.Results The excellent or good image quality (score 1 and 2)of the four groups were above 95%,and no statistical differences in images scores was detected in A1 and B1 (P =0.726),A2 and B2 (P =0.079).Effective dose was significantly different in A1 and B1(P <0.001),A2 and B2 (P <0.001),and the former decreased 50% and 40% as compared with the latter.Conclusion Using prospective ECG gated scan and adaptive cardio sequence scan,assessable image quality of coronary artery can be acquired with dual source CT (heart rate<90 beast per minute),and radiation dose can be obviously reduced.

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region.The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis.The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the ?-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.

Objective The purpose of this study was to evaluate the safety and clinical oulcomes of aortic valve replacement (AVR) performed with minimally invasive technique.Methods From June 2010 to October 2011,20 cases of minimally invasive AVR were performed.The mean age was (47.60±12.28) ;12 males and 8 females.All patients are ventilated with a double-lumen endotracheal tube,through the 3nd anterior intercostals space with a 5 -6cmskin incision,right femoral artery and vein cannulation are used to establish CPB,direct aortic cross-clamped by Chitwood sliding clamp through the right 4th intercostals space,and completed the aortic valve replacement.Results Mean length of incision was (4.73±0.54)cm.Mean duration of cardiopulmonary bypass was (124±39.83)min,crossclamp time was (97.21±33.17) min.Median intubation time was (13.55±3.87)hours.Median duration of intensive care and postoperative hospital stay was (16.34±3.82)hours and (6.63±1.45) days,respectively.Hospital mortality was 0.There was no perivalvular leakage,Conclusion Minimally invasive aortic replacement with a modified Port-Access approach is feasible,small incisions,more cosmetic,shorter length of bospital stay and less need for blood transfusion are attainable.

Objective To investigate the expression of extraeellular signal-regulated protein kinase (ERK1/2)pathway in rat brains with fluorosis and the effects of fluoride on learning and memory of the rats,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of the ion.Methods Seventy-two SD rats were divided into 3 groups and 24 rats were in each group.Three groups were fed respectively with different concentrations of fluoride(NaF)for 6 months to establish rat models with fluorosis.Controls were fed with tap water (NaF＜0.5 mg/L):lower and higher concentration group were fed with water containing NaF(5,50 ms/L).Animals are sacrificed after 6 months of treatment with fluoride and the dissected brains were kept for analysis.The protein levels of ERK1/2 in rat brains were detected by Western-blotting and the mRNA level by RT-PCR. The spatial learning and memorizing ability was measured by Morris water maze test. Results The ERK1/2 protein in control group,lower and higher concentration group was 0.944±0.10,1.253±0.02,1.953±0.07,the differece being statistieally sighificant between any two groups (P ＜ 0.05). The phospho-ERKl/2 protein in control group,lower and higher concentration group was 0.73±0.08,0.77±0.07,1.28±0.11,the differece being statistieally sighificant between any two groups(P ＜ 0.05);the activation rate of phospho-ERK1/2 in lower and higher concentration group [(68.4± 3.8)%,(64.1±3.2)%] was decreased compared to control group[ (82.3±10.7)%],the differece being significant(P ＜ 0.05). In the navigation trial,longer escape latencies of lower concentration group on the second, the third,the fifth and the sixth day were observed[ (46.0±8.0),(24.0±2.7),(8.9±5.3),(7.4±4.1 )s] compared to the control[ (39.3±6.9),(19.1±9.1 ),(8.3±3.4),(4.8±2.7)s],the differece being significant (P ＜ 0.05 or ＜ 0.01 );the similar results were also observed in the higher concentration group[ (36.9±16.8),(37.7±12.9), (19.7±7.6),(12.2±5.7 )s],and the escape latencies of the higher concentration group on the third,the fifth and the sixth day were longer than that in lower concentration group. In the probe test,the rats took more time to reach the first cross in lower and higher concentration group[(1.17±0.75),(4.18±1.10)s] than control group[ (5.89± 0.56 ) s ],the differece being significant (P ＜ 0.05 or ＜ 0.01 ) ;stayed shorter [ ( 17.05±4.25 ),(18.20±4.57 ) s ] than control [(25.37±5.65 )s ] in platform area (P ＜ 0.01 );the activation rates of ERK1/2 were directly correlated with the time taken to reach the first cross platform located in the probe test(r = 0.364,P ＜ 0.05) and the activation rates were also directly correlated with the escape latencies on the sixth day(r = 0.497,P ＜ 0.05). Conclusion Long-term exposure of excessive fluoride induces the change of expression and activating rate of the ERK1/2 in rat brains,leading to the decreased capacity of learning and memory.

Objective To investigate the expression and distribution of the downstream substrate of extracellular regulated protein kinase(ERK1/2) pathway, ternary complex factor phospho-Elk-1, in rat brains with chronic fluorosis, and reveal the mechanism of the impaired learning and memory ability caused by chronic fluorosis. Methods Seventy-two SD rats, weighing 100 - 120 g, were randomly divided into 3 groups, 24 in each group (half male and half female). The rats in control group were fed with tap water (fluoride < 0.5 mg/L); low- and high-dose fluoride groups were fed with tap water with different concentrations of NaF(5.0,50.0 mg/L F-, respectively). After 6 months, body weight was weighed, dental fluorosis was determined by observation and urinary fluoride and bone fluoride were detected by fluorine ion-selective electrode; the learning ability of rats was measured by navigation test of Morris water maze, and memory ability by spatial probe test in Morris water maze; the expression and distribution of phospho-Elk-1 in different brain regions were detected by immunohistochemistry method. Results In low- and high-fluoride groups, the body weight of rat[(449.2 ± 77.1), (312.8 ± 89.7)g] was significantly decreased than that of control [(635.5 ± 76.2 )g, all P< 0.05], the varying degrees of dental fluorosis were observed(x2 = 7.83, P<0.05), urinary fluoride[(2.56 ±0.91),(5.73 ±3.14)mg/L] and bone fluoride[(709.2 ± 37.4) ,(1306.3 ± 102.4) mg/kg] were significantly higher than those in controls[(0.92 ± 0.30)mg/L,(348.5 ± 89.2)mg/kg, all P< 0.05]. The escape latency of low- and high-fluoride groups[ (7.4 ± 4.1), (12.2 ± 5.7)s] was longer than that of control [(4.8 ± 2.7 )s, all P < 0.05] and the escape latency in high-fluoride group was significantly longer than that in other groups (all P < 0.05); in spatial probe test, the time of first crossing platform was longer in rats with fluorosis [(4.18 ± 1.10),(5.89 ± 0.56)s] as compared to control[(1.17 ± 0.75)s, all P< 0.05]. Expressions of phospho-Elk-1 in the hippocampus CA1(167.4 ± 8.3,163.2 ± 9.4), CA2(175.7 ± 5.0,183.3 ± 4.2), CA3(165.2 ± 11.6,162.9 ± 4.4), CA4(168.7± 6.9,169.5 ±5.3), fascia dentate (185.2 ±4.0,193.1 ±6.1) and caudate putamen( 181.4 ± 3.8, 179.8 ± 5.5) in low- and high-fluoride groups were higher than those of controls(142.4 ± 8.1,144.9 ± 8.4,143.6 ± 5.8, 116.8 ± 9.1,140.2 ± 7.8,163.1 ± 13.1, all P< 0.05). Conclusion Chronic fluorosis can cause increased expression of phospho-Elk-1 in the hippocampus and caudate putamen region of rat brains, which might be related to the mechanisms of decreased learning and memory ability of rats overexposed to fluoride.

Objective To investigate the changes of oxidative stress level in brain tissues and serum, and learning and memory in rats with oxidative stress level in nerve damage in chronic fluorosis. Methods The rats were randomly divided into 3 groups according to the body weight, eight rats in each group, i.e., control group, drinking water containing less than 0.5 mg/L of fluoride; lower fluoride exposure group, drinking water containing 5 mg/L of fluoride; higher fluoride exposure group, drinking water containing 50 mg/L of fluoride. The animals were examined six months after initiating the experiment. The total antioxidant capacity (T-AOC) and malondialdehyde (MDA), as well as learning and memory, were measured. Results Escape latency in higher fluoride exposed group[ (14.37±3.48)s] was significantly higher than that of controls[ (5.84±1.87)s] and exposed te lower fluoride [ (7.18±1.42)s], the difference being statistically signifieant(P<0.05). As compared with controls[ (2.17±0.11)× 103 U/L , (0.79±0.11)×103 U/g Pr] ,the rats exposed to higher fluoride and lower fluoride exhibited lower levels of T-AOC [(1.37±0.27)×103 U/L,(0.24±0.06)×103 U/g Prand (1.20±0.14) x 103 U/L,(0.41 ~ 0.10)×103 U/g Pr], the difference being statistically signifieant(P<0.05). As compared with controls[ (2.34±0.16) mmoL/L, (2.97±0.11)mmol/g Pr] and low fluoride exposed group[ (2.68±0.33)mmoL/L, (3.38±0.21)mmol/g Pr], higher level of MDA were observed in higher fluoride exposed group[ (3.72±0.59)retool/L, (4.01±0.21)mmol/g Pr], the difference being statistically significant(P<0.05). Conclusion The results indicated that higher amount of fluoride induced an increased level of oxidation, which might result in the decreased capacity of intelligence of rats with fluorosis.

Objectives To summarize the characteristics,differential diagnosis and management of incomplete 17 alpha-hydroxylase/17,20-1yase deficiency(17 OHD)of Chinese patients.Methods Six cases of incomplete 17 OHD from Peking Union Medical College Hospital were studied retrospectively through analyzing their clinical data,and the molecular pathogenic mechanism was discussed after literature review.Results Four cases of 46,XX incomplete 17 OHD were reported.The clinical characteristics included female phenotype,various degrees of breast development and absent or sparse axillary/pubic hair, oligomenorrhea or secondary amenorrhea,recurrent luteinized ovarian cysts,hypogonadism with persistent hyperprogesteronemia or high serum 17 alpha-hydroxyprogesterone level,with or without hypokalemic hypertension.There were also 2 cases of 46,XY incomplete 17 OHD,in which ambiguous genitalia were present besides hypokalemic hypertension.Conclusions Incomplete 17 OHD is a very rare form of congenital enzymatic deficiencies of steroid synthesis,which should be included in the differential diagnosis when there are menstrual disorders,sexual infantilism,recurrent ovarian cysts or ambiguous genitalia.Under such circumstances,hyperprogesteronemia offers a valuable clue for further investigation.

BACKGROUND:With the medical development, prognostic outcomes of spinal cord injuries have not been improved significantly, and most patients also suffer from severe complications. Nowadays, lots of laboratories and clinical researches have suggested that celltherapy has a great potential, especial y the application of umbilical cord blood stem cells in nervous system diseases. OBJECTIVE:To explore the feasibility and clinical effect of umbilical cord blood neural stem cells transplantation for patients with obsolete spinal cord injury. METHODS:Umbilical cord blood was harvested from newborns under aseptic condition, and differentiated into neural stem cells in vitro that were prepared into cellsuspension at a concentration of 109/L. The cellsuspension (3 mL) was injected via the L 3-4 or L 4-5 into the subarachnoid space. American Spinal Injury Association (ASIA) scores and the residual urine were assessed before and 3 months after transplantation. RESULTS AND CONCLUSION:After transplantation, al the patients showed a stable life indication. Three months later, ASIA scores were increased and the residual urine decreased, which significantly differed from those before transplantation (P<0.05). These findings indicate that umbilical cord blood neural stem cells transplantation is a new treatment that can improve the limb function and life quality of patients with obsolete spinal cord injury.