A high sensitive epitope imprinted electrochemical sensor for bovine serum albumin based on enzyme amplifying.

To improve the sensitivity of the molecular imprinting sensor detection of protein, a new strategy based on enzyme amplification was proposed. The determination of bovine serum albumin (BSA) was achieved by using the epitope imprinted techniques coupling with electrochemical measurement method. Nonapeptide, separated from BSA, was selected as a template molecule to prepare the molecularly imprinted polymer (MIP) film, and it could bind with the cavities of the MIP. By the use of epitope imprinted techniques, BSA can be recognized by the MIP via the nonapeptide on the surface of BSA. The synthesized horseradish peroxidase-labeled nonapeptide (HRP-nonapeptide) can also be recognized by the MIP. After the competitive reaction between HRP-nonapeptide and BSA, the enzymatic reaction derived from labeled HRP on the H2O2-hydroquinone system make the electrochemical current of hydroquinone change, then the concentration of BSA can be indirectly determined. BSA in the range of 1.0-150 ng/mL exhibited a linear relationship with the differential pulse voltammetric current variation and the detection limit was 0.02 ng/mL. The sensor has high sensitivity, good selectivity, and reproducibility. It has been applied to the determination of residual bovine serum albumin in human rabies vaccine with the recovery rate of 98.3%-102.5%.

Misdiagnosis of a small cell lung cancer resulting from inaccurate pathology.

Currently, a biopsy provides the most reliable evidence for diagnosing a disease, and the majority of doctors do not question the diagnosis made by a pathologist. However, an inaccurate diagnosis may lead to serious consequences; for example, a benign tumor may be misdiagnosed as a malignancy, or a malignancy may be deemed to be benign. How to avoid these types of mistakes is a continuing issue of concern to all doctors. Here, we report a case of small cell lung cancer misdiagnosed as an inflammatory myofibroblastic tumor. Fortunately, we performed a mediastinoscopy on the patient and discovered the actual pathologic condition. This case is presented to caution against the possibility of the misdiagnosis of uncommon diseases in clinical practice.

Aberrant Expression of HOXA5 and HOXA9 in AML.

BACKGROUND: Aberrant expression of HOX gene expression has been observed in cancer. The purpose of this study was to investigate the alteration of HOXA5 and HOXA9 expression and their clinical significance in acute meloid leukemia (AML). MATERIALS AND METHODS: The expression of HOXA5 and HOXA9 genes of bone marrow samples from 75 newly diagnosed AML patients and 22 healthy controls for comparison were examined by Real- time quantitative PCR (RQ-PCR) assay. Statistical analysis was conducted to evaluate HOXA5 and HOXA9 expression as possible biomarkers for AML. RESULTS: The results showed that the complete remission rate (52.6%) of the patients who highly expressed HOXA5 and HOXA9 was significantly lower than that (88.9%) in patients who lowly express the genes (P=0.015). Spearmann correlation coefficients indicated that the expression levels for HOXA5 and HOXA9 genes were highly interrelated (r=0.657, P<0.001). Meanwhile, we detected significant correlations between HOXA9 expression and age in this limited set of patients (P=0.009). CONCLUSIONS: The results suggest a prognostic impact of increased expression of HOXA5 and HOXA9 in AML patients.

[Effect of dietary vitamin A intake on plasma vitamin A concentration in preschool children of Banan district, Chongqing, China].

OBJECTIVE: To study the relationship between dietary vitamin A intake and plasma vitamin A concentration, and establish the theoretical basis for dietary intake predicting vitamin A nutritional status. METHODS: By using cluster sampling, 492 children aged 2-7 years in kindergartens in Banan district of Chongqing were selected. A cross-sectional nutrition and health survey was conducted, including the clinical examination, anthropometry, laboratory test and dietary survey. RESULTS: Among the children surveyed, 229 were boys, and 263 girls, the mean age was (4.54 ± 0.87) years, height (107.50 ± 7.20) cm, and weight (18.42 ± 3.41) kg, the mean value of plasma vitamin A was (1.04 ± 0.30) µmol/L. The prevalence of marginal vitamin A deficiency (MVAD) was 43.5%. No cases of severe clinical vitamin A deficiency were found (plasma vitamin A ≤ 0.35 µmol/L). Clinical examination found no conjunctiva, corneaor skin abnormalities, and no Bitot's spots. Prevalence of the last two weeks colds were 27.4% (135/492), no diarrhea and other gastrointestinal or digestive diseases were found. The proportion of insufficient dietary vitamin A intake (<600 µg RE/d) was as high as 50.0%. By using correlation analysis, plasma retinol concentrations were related to dietary vitamin A intake (r=0.162, P<0.001), and to dietary energy intake (r=0.107, P=0.017). After adjustment for the effects of other non-dietary factors on vitamin A deficiency, the multivariate logistic regression showed that vitamin A-rich foods of liver intake=0 g/d (OR=1.95, 95% CI: 1.05-3.61, P=0.034), vitamin A-rich fruits intake=0 g/d (OR=1.55, 95% CI: 1.03-2.33, P=0.034), vitamin A-rich vegetables intake<200 g/d (OR=3.47, 95% CI: 1.37-8.75, P=0.009) were important risk factors of vitamin A deficiency. But we had not found the correlation between the intake of meat, eggs and milk and vitamin A deficiency. CONCLUSION: Dietary factors may be the major risk factor of vitamin A deficiency in the three kindergartens. The dietary vitamin A intakes are significantly related to plasma retinol concentrations, and the vitamin A-rich foods intakes can predict the body's vitamin A nutritional status.

Delayed gastric emptying after esophagectomy for malignancy.

BACKGROUND: Controversy still exists about the need for pyloric drainage procedures after esophagectomy with gastric conduit reconstruction. Although pyloric drainage may prevent postoperative delayed gastric emptying (DGE), it may also promote dumping syndrome and bile reflux. The aims of this study were to audit the incidence and management of DGE in patients without routine pyloric drainage after esophagectomy in a university medical center. PATIENTS AND METHODS: From July 2006 to June 2012, data from 356 consecutive patients who underwent esophagectomy with a gastric conduit without pyloric drainage for esophageal or gastric cardia carcinoma were reviewed. Major observation parameters were the incidence, management, and outcomes of DGE. RESULTS: Overall incidence of DGE was 15.7% (56 of 356). Early DGE developed in 26 patients, and late DGE developed in 30 patients. There were no differences in demographic and intraoperative data between the two groups with or without DGE. More DGE was documented in patients with an intra-right thoracic gastric conduit (P=.031). A higher incidence of postoperative pneumonia was observed in patients exhibiting early DGE, but without significance (P=.254). There were also no significant impacts on respiratory failure (P=.848) and anastomotic leakage (P=.257). There was an increased postoperative hospital stay with DGE, but without significance (P=.089). Endoscopic balloon dilatation of the pylorus was used to manage 33.9% of patients with DGE, yielding a 78.9% (15 of 19) success rate without complications. In 3 patients endoscopy showed the pylorus was open, and their symptoms improved over time. One patient with tumor-related DGE was treated by pyloric stent. The remaining patients were adequately treated with conservative management. CONCLUSIONS: Omitting the operative drainage procedure does not lead to an increased frequency of DGE after esophagectomy with a gastric conduit. Many patients responded to conservative management, and endoscopic balloon pyloric dilatation can be effective in managing the DGE postoperatively.

Vitamin A deficiency impairs postnatal cognitive function via inhibition of neuronal calcium excitability in hippocampus.

Vitamin A (VA) is important for postnatal brain development, and VA deficiency (VAD) can cause learning and spatial memory deficits in rats. Most of the biological functions of VA are mediated by retinoic acid (RA). To investigate the mechanisms underlying VA deficits, mother rats were fed elemental diets to achieve blood VA levels classified as normal, deficient or severely deficient. Shuttle box and Morris water maze tests revealed impairments in learning ability and spatial memory, respectively, in adolescent VAD rats (p 30-35). Electrophysiology showed weaker long-term potentiation in VAD rats compared to VA normal rats. Examination of NMDA-induced calcium (Ca(2+) ) excitability revealed decreased excitability in hippocampal slices from VAD rats during postnatal development. Relative to VA normal rats, VAD rats also had decreased NMDA receptor NR1 mRNA and protein expression in later stages of postnatal development (p 10-30), as well as differences in retinoic acid receptor (RARα) mRNA and protein expression. Furthermore, primary hippocampal neurons in culture showed increased neuronal Ca(2+) excitability in response to all-trans-RA or 9-cis-RA, coupled with increases in RARα and NR1 expression similar to those observed in vivo. We also found weaker calcium excitability and lower expression of NR1 mRNA and protein after specific silencing of RARα. Finally, we found that RA signals affected the expression of NR1 do not directly through transcriptional regulation. These data support the new idea that continuous postnatal VAD inhibits RARα expression, which decreases NR1 expression via no direct transcriptional regulation and then inhibits hippocampal neuronal Ca(2+) excitability which affects long-term potentiation, finally producing deficits in active learning and spatial memory in adolescence.

[Impact of antioxidant vitamins and heavy metal levels at birth on neurodevelopment of children assessed at two years of age].

OBJECTIVE: Epidemiologic study on the perinatal interaction of antioxidant vitamins and heavy metals on offspring's intellectual development is extremely limited. The aim of this prospective cohort study was to investigate the status of maternal and cord blood antioxidant vitamins (Vitamin A, E and C) and heavy metals (lead, cadmium and mercury) at delivery and correlations between these antioxidant vitamins or metals and neurodevelopment of early childhood, and to explore the protective effect of antioxidant vitamins against the injure of heavy metals to intellectual development in children in Tongliang, Chongqing, China. METHOD: A total of 150 pairs of mothers-neonates were recruited. Serum concentrations of vitamin(V) A, vitamin E, vitamin C, lead, cadmium and mercury in maternal and cord blood after delivery were determined and intellectual development was evaluated by Gesell Development Schedule (GDS) at two years of age. RESULT: A total of 111 pairs of maternal-neonatal subjects with completed data were included into statistical analysis. After adjusting for potential confounders, cord blood mercury level was found negatively associated with all the development quotients (DQs) (ß = -0.486, ß = -0.716, ß = -0.846, ß = -0.935, ß = -0.702, P < 0.05). Cord VE level was positively correlated with motor, adaptation, language and average DQ (ß = 0.475, ß = 0.458, ß = 0.403, ß = 0.395, P < 0.05). When antioxidant vitamins and heavy metals levels were all included into the multiple regression model, cord vitamin E level was statistically positively related with motor, adaptation and average DQs (ß = 0.449, ß = 0.412, ß = 0.349, P < 0.05). While cord blood mercury level was negatively correlated with adaptation, language, social and average DQ (ß = -0.397, ß = -0.727, ß = -0.935, ß = -0.628, P < 0.05), the relationship between cord mercury level and motor DQ was not statistically significant (P = 0.0890). The motor, adaptation, language and average DQs in high cord VE group were higher than those in low VE group (t = 2.93, t = 2.06, t = 2.13, t = 2.27, P < 0.05). Social DQ in high cord lead group was significantly lower than that in low lead group (χ(2) = 5.56, P = 0.015). Cord VC level in high cadmium group was significantly lower than that in low cadmium group (χ(2) = 7.62, P = 0.006). VA placental transport ratio in high mercury group was significantly lower than those in low mercury group (χ(2) = 8.02, P = 0.005). CONCLUSION: Our data suggested that there may be certain interaction between antioxidant vitamin A, E, C and heavy metals lead, cadmium, mercury. Antioxidant vitamins at delivery may have protective effect on the early neurodevelopment via interaction with the heavy metals.

The pattern of retinoic acid receptor expression and subcellular, anatomic and functional area translocation during the postnatal development of the rat cerebral cortex and white matter.

Retinoic acid (RA), which is an important modulator of brain development, neural cell proliferation, neurite outgrowth, and synaptic plasticity, is regulated via changes in RA receptors. The pattern of RA receptor changes in the rat cerebral cortex and white matter during postnatal development has not been extensively studied. Therefore, we studied the mRNA expression patterns of 6 RA receptors in the postnatal rat cerebral cortex and white matter at 1, 3, 7, 10, 14, 21, 28, and 35days. We found that RARß, RXRα and RXRß mRNA levels gradually increased during postnatal development. RARα presented a nearly unimodal trend, but RARγ and RXRγ were generally bimodal. RARα, RARγ, and RXRγ mRNA levels peaked at postnatal day 21 (P21). The pattern of RARα expression was consistent with that of its mRNA levels. RARα and RXRγ mRNA levels were the highest among those of all RA receptors during postnatal development. Interestingly, RARα expression was mainly located in the cytoplasm in the postnatal development apart from P3d. We further showed that RARα is expressed mainly in layers 2 and 3, partly in layers 1 and 4, and in a limited manner in layers 5 and 6 in the parietal cortex. Most RARα expression occurs in layers 2, 3, and 4 in the temporal lobe cortex. We realized that the M1 S2 region of RARα is highly expressed and that the position of RARα changes dynamically to meet the needs of different regions during development. These results support the idea that the RA receptor plays an important role in the cerebrum during postnatal development and implementation of these functions may be mainly dependent on the non-transcriptional or post- transcriptional regulation.

Effect of vitamin A, vitamin A plus iron and multiple micronutrient-fortified seasoning powder on infectious morbidity of preschool children.

OBJECTIVE: Improvement of hemoglobin and serum retinol and facilitation of the mobilization of iron storage were achieved with a multiple-micronutrient-fortified diet in preschoolers for 6 mo in a suburb of Chongqing, China. We investigated whether fortification with multiple micronutrients in a diet for preschool children results in changes in children's infectious morbidity compared with diets fortified solely with vitamin A and with vitamin A plus iron. METHODS: From December 2005 to June 2006, 226 2- to 6-y-old preschool children were recruited from three nurseries randomly assigned to three different fortified-diet groups for 6 mo. Group I was fortified with vitamin A; groups II and III were fortified with vitamin A plus iron and vitamin A plus iron, thiamine, riboflavin, folic acid, niacinamide, zinc, and calcium, respectively. The secondary functional outcomes, morbidity of diarrhea and respiratory infection, were collected during supplementation. RESULTS: The groups were comparable concerning compliance and loss to follow-up. There was evidence of a lower incidence rate of respiratory-related illnesses, diarrhea-related illness, fewer symptoms of runny nose, cough, and fever, and shorter duration of respiratory-related illnesses and cough for children in group III compared with children in groups I and II. However, there was no significantly or clinically important difference between children in groups I and II. CONCLUSION: The beneficial effects on infectious morbidity over 6 mo, in addition to some biochemical improvements, highlight the potential of this micronutrient-fortified seasoning powder supplied in a diet for preschool children.

[Primary culture of rat hippocampal neurons and detection of the neuronal excitability].

OBJECTIVE: To improve the efficiency of primary culture of hippocampal neurons and obtain highly purified neurons with good in vitro growth and minimal risk of contamination. METHODS: The hippocampal neurons of neonatal Wistar rats were isolated and the single cell suspension was prepared by mechanical trituration and sedimentation in stead of trypsin digestion and filteration. Twenty-four hours after the cell plating, the culture medium was removed and replaced by serum-free DMEM/F12 with B27 supplementation. Half of the culture medium was changed 2-3 times every week. The morphological changes of the neurons were observed under inverted phase-contrast microscope. Immunofluorescence staining for NSE was performed to identify the neurons, and the purity of neurons was calculated. The hippocampal neurons were stained with calcium-sensitive fluorescent dye to monitor the effect of KCl on neuronal excitability by a calcium imaging system. RESULTS AND CONCLUSION: This simplified method is time-saving and cost-effective for primary culture of hippocampal neurons with reduced risk of contamination, and the neurons obtained showed high uniformity, purity and long-term viability.

Antioxidant vitamin status during pregnancy in relation to cognitive development in the first two years of life.

OBJECTIVE: To investigate the correlation of the antioxidant vitamins status (vitamins A, E and C) during pregnancy and the intellectual development of early childhood. METHOD: A total of 150 paired maternal-neonatal subjects were recruited into the present study. The serum concentrations of antioxidant vitamins (vitamins A, E and C) in maternal blood and cord blood after delivery were determined by high performance liquid chromatography and the intellectual development was evaluated by Gesell Development Schedules (GDS) at two-years-old. RESULT: Children with higher cord serum vitamin E level showed higher scores of motor, adaptive domain and average compared to children with lower cord serum vitamin E level (p<0.01 or 0.05), respectively. Cord serum vitamin A level had significant positive correlation with effect on motor DQs (beta=4.227, p<0.05), and vitamin E level in cord blood showed a positive relation with motor DQ and average DQ (beta=0.329 and 0.1875, respectively, p<0.05) in multiple linear regression model. The language and social DQs were influenced by placental vitamin E transport rate (beta=3.1968 and 3.0194, respectively, p<0.05). The placental transport rate of vitamin E also was a protective factor for the prevalence of motor behavior developmental delay [OR: 0.118, 95% confident interval (95% CI), 0.018-0.765, p=0.0251], personal and social behavior developmental delay (OR: 0.052, 95% CI: 0.004-0.610, p=0.0185) and average developmental delay (OR: 0.041, 95% CI: 0.003-0.642, p=0.0229) in logistic multiple regression model. CONCLUSION: Data suggested that vitamin A, E status and vitamin E transfer rate at delivery had beneficial influence on children's cognitive and behavior development quotients.

Perinatal vitamin A status in relation to neurodevelopmental outcome at two years of age.

UNLABELLED: Information about the effect of antioxidant vitamins nutrition during pregnancy on offsprings intellectual development is extremely limited. OBJECTIVE: To investigate the correlation of antioxidant vitamins (Vitamin A, E and C) at delivery and the neurodevelopment of early childhood. METHOD: A total of 158 paired maternal-neonatal subjects were recruited. The serum concentrations of vitamin A, E and C in maternal and cord blood after delivery were determined and intellectual development was evaluated by Gesell Development Schedule (GDS) at two years old. RESULT: After adjusting for potential confounders, vitamin A placental transport ratio (VA-PTR) was positively associated with motor area development quotients (DQ) and average DQ(p<0.01). Cord VA level was positively related with language area and social area DQ (p<0.05). Nevertheless, there was no significant association between cord VE, VC levels, VE PTR or VC PTR and GDS. The adaptive area and average DQ in high cord VA group was higher than those in low VA group (p<0.05). Cord VA level and VA-PTR were positively associated with birth head circumference and birth weight, respectively. CONCLUSION: Our data suggested that adequate vitamin A at delivery had beneficial influence on neonatal birth outcomes and children's neurodevelopment in later childhood.

[Clinical observation of XELOX (Capecitabine puls Oxaliplatin): an adjuvant chemotherapy regimen used in stage III colorectal cancer].

OBJECTIVE: To evaluate the efficacy and safety of an adjuvant chemotherapy regimen: XELOX (Capecitabine puls Oxaliplatin) used after curative resection for stage III colorectal cancer. METHODS: From Jan. 1998 to Jan. 2004, 256 cases with stage III colorectal cancer randomized received de Gramont, modified FOLFOX4 (mFOLFOX4) and XELOX regimens. The 3-year disease-free survival (DFS) and overall survival (OS) were compared within the three groups and relative prognosis factors within mFOLFOX4 and XELOX groups. Therapeutic adverse events were recorded and analyzed with Kaplan-Meier test. RESULTS: 98, 87 and 71 cases were respectively enrolled in the de Gramont, mFOLFOX4 and XELOX groups, mFOLFOX4 and XELOX had superior efficacy compared with de Gramont regimen. The two former could significantly improve 3-year DFS (79.7% vs. 66.2%, P = 0.015; 81.5% vs. 66.2%, P = 0.004) and medium survival time (40.2 mon vs. 37.8 mon, P = 0.024; 41.4 mon vs. 37.8 mon, P = 0.014). Meanwhile they could respectively decrease the ratio of recurrence risk by 18.0% (P = 0.024) and 21.0% (P = 0.003). The relative benefit of mFOLFOX4 versus XELOX didn't differ for 3-year DFS [hazard ratio (HR): 0.84, 95% confidence interval (CI): 0.79-1.12, P = 0.13] and OS (HR: 0.87, 95% CI: 0.84-1.06, P = 0.54). In the analysis of DFS in relative prognosis factors, XELOX had a better trend of survival advantage. mFOLFOX4 had higher adverse events within these regimens, especially in grade 3 or 4 neutropenia and peripheral neurologic adverse events. CONCLUSION: XELOX maintains its efficacy and safety ratio in advanced colorectal cancer. Patients have good tolerance and compliance. The regiment is deserves to be applied in clinical treatment. Oxaliplatin;

[Karyotypes and pelvic ultrasonography in children with Turner syndrome].

OBJECTIVE: To study the relationship of karyotypes and gonadal development in children with Turner syndrome. METHODS: Fifty-one children with Turner syndrome were studied. Pelvic ultrasonography was performed on the children. Twenty healthy age-matched girls were used as control group. RESULTS: Eleven kinds of karyotypes were detected in 51 children with Turner syndrome. Children were classified into two groups based on karyotypes: Group 1 (45,XO; n= 24) and Group 2 (karyotypes other than 45, XO; n=27). 45,XO karyotype was the most common (47.1%), followed by 46,X,i (Xq)/45, XO (21.6%), 46,XX/45, XO (7.8%) and 47,XXX/45, XO (5.9%). Pelvic ultrasonography showed a primordial uterus in 41 cases, infantile uterus in 7 cases, congenital absence of uterus and ovary in 3 cases, simple anovarism in 42 cases and ovarian dysgenesis in 6 cases. Uterine size in both Groups 1 and 2 were significantly smaller than those of the control group (P<0.05). Group 1 showed smaller size of uterus than Group 2 (P<0.05). Ovaries were not detected in all Group 1 patients. CONCLUSIONS: The dysplasia and atrophy of ovaries and uterus exist in children with Turner syndrome. The patients with 45,XO karyotype had poorer gonadal development than those with other karyotypes.

Donor denervation and elimination of Kupffer cells affect expression of P-selectin and ICAM-1 in liver graft.

BACKGROUND: The non-function and dysfunction of primary liver graft likely involves dependence on Kupffer cells and hepatic innervation. The present experiment was designed to study the expression of P-selectin and intercellular adhesion molecule-1 (ICAM-1) mRNA in liver graft and to elucidate the role of Kupffer cells and the sympathetic nerve of the liver in down-regulating this expression. METHODS: Donor rats were given hexamethonium, a sympathetic ganglionic blocking agent, and/or gadolinium chloride, an inhibitor of Kupffer cells. Then the changes of graft P-selectin and ICAM-1 mRNA expression were measured after liver transplantation. RESULTS: The expressions of P-selectin and ICAM-1 mRNA were increased after liver transplantation, and down-regulated by liver denervation and elimination of Kupffer cells. CONCLUSIONS: Live donor denervation and elimination of Kupffer cells down-regulated the expressions of P-selectin and ICAM-1 mRNA in grafts. This may decrease graft ischemia/reperfusion injury.