[Clinical features and gene mutations of 6 patients with carnitine palmitoyltransferase 1A deficiency].

The clinical and biochemical data and gene sequencing results of patients with carnitine palmitoyltransferase 1A deficiency were analyzed, in order to improve the understanding of the disease. Six patients (5 males and 1 female, aged from 1 to 8 years old) with carnitine palmitoyltransferase 1A deficiency from Department of Pediatric Endocrinology and Genetic Metabolism, Xinhua Hospital between 2008 and 2019 were included. Two cases were detected by neonatal screening and had no clinical symptoms. The remaining 4 cases all showed seizures induced by fever, vomiting or diarrhea. All the 6 patients showed increased serum free carnitine (C0), decreased hexadecanoylcarnitine (C16) and octadecanoylcarnitine (C18), and increased C0/(C16+C18). Meanwhile, compound heterozygous mutations of CPT1A gene were detected in all 6 patients, of which 2 were reported mutations (c.281+1G>A and c.968-8C>T), and 10 were new mutations. The new mutations included 6 missense mutations, 1 nonsense mutation, 1 deletion mutation and 2 splicing mutations. Detection of free carnitine and acyl carnitine by tandem mass spectrometry is helpful for early screening and diagnosis of carnitine palmitoyltransferase 1A deficiency.

Plasma metabolite changes in dairy cows during parturition identified using untargeted metabolomics.

The metabolic responses of cows undergo substantial changes during the transition from late pregnancy to early lactation. However, the molecular mechanisms associated with these changes in physiological metabolism have not been clearly elucidated. The objective of this study was to investigate metabolic changes in transition cows from the perspective of plasma metabolites. Plasma samples collected from 24 multiparous dairy cows on approximately d 21 prepartum and immediately postpartum were analyzed using ultra-high-performance liquid chromatography/time-of-flight mass spectrometry in positive and negative ion modes. In conjunction with multidimensional statistical methods (principal component analysis and orthogonal partial least squares discriminant analysis), differences in plasma metabolites were identified using the t-test and fold change analysis. Sixty-seven differential metabolites were identified consisting of AA, lipids, saccharides, and nucleotides. The levels of 32 plasma metabolites were significantly higher and those of 35 metabolites significantly lower after parturition than on d 21 prepartum. Pathway analysis indicated that the metabolites that increased from late pregnancy to early lactation were primarily involved in lipid metabolism and energy metabolism, whereas decreased metabolites were related to AA metabolism.

Evaluating the potential of a new isotope-labelled glyco-ligand for estimating the remnant liver function of schistosoma-infected mice.

A new glyco-derivative compound (OCTAM) was developed and labelled with isotope to form (188) Re-OCTAM as a candidate nuclear medicine imaging agent for testing the liver function. We evaluated the potential of isotope-labelled OCTAM for estimating the remnant liver function in vitro and in vivo schistosoma-infected mice. The affinity of OCTAM to liver asialoglycoprotein receptors (ASGPR) was assessed by competitive inhibition assay in vitro. In vivo assessments were performed to score the remnant liver function in mice at different schistosomal infection stages. OCTAM binds specifically to ASGPR and showed competitive inhibition of anti-ASGPR antibody binding to hepatocytes, and was higher than that of other galactosyl ligands. Micro-SPECT/CT images of uninfected mice revealed strong liver uptake. Quantified serial images of mice infected for 9, 12 and 18 weeks showed delayed liver uptake, and the retention of uptake was inversely correlated with stage and grade of schistosoma infection. Pathological and biochemical analysis demonstrated that gradually accumulating liver injury caused by infection significantly influenced uptake of (188) Re-OCTAM. Hepatic ASGPR expression diminished only in the chronic infection stage. This study demonstrated that the isotope-labelled OCTAM could accumulate in the liver, might have potential as an imaging agent for in vivo hepatic function evaluation of schistosomiasis.

Astrocytic endogenous glial cell derived neurotrophic factor production is enhanced by bone marrow stromal cell transplantation in the ischemic boundary zone after stroke in adult rats.

Bone marrow stromal cells (BMSCs) facilitate functional recovery in rats after focal ischemic attack. Growing evidence suggests that the secretion of various bioactive factors underlies BMSCs' beneficial effects. This study investigates the expression of glial cell derived neurotrophic factor (GDNF) in the ischemic hemisphere with or without BMSC administration. Adult male Wistar rats were subjected to 2 h of middle cerebral artery occlusion followed by an injection of 3 x 10(6) BMSCs (n = 11) or phosphate-buffered saline (n = 10) into the tail vein 24 h later. Animals were sacrificed seven days later. Single and double immunohistochemical staining was performed to measure GDNF, Ki67, doublecortin, and glial fibrillary acidic protein expression as well as the number of apoptotic cells along the ischemic boundary zone (IBZ) and/or in the subventricular zone (SVZ). BMSC treatment significantly increased GDNF expression and decreased the number of apoptotic cells in the IBZ (P < 0.05). GDNF expression was colocalized with GFAP. Meanwhile, BMSCs increased the number of Ki-67 positive cells and the density of DCX positive migrating neuroblasts (P < 0.05). GDNF expression was significantly increased in single astrocytes collected from animals treated with BMSCs, and in astrocytes cocultured with BMSCs after OGD (P < 0.05). Our data suggest that BMSCs increase GDNF levels in the ischemic hemisphere; the major source of GDNF protein is reactive astrocytes. We propose that the increase of GDNF in response to BMSC administration creates a hospitable environment for local cellular repair as well as for migrating neuroblasts from the SVZ, and thus contributes to the functional improvement.

Association of vitamin D binding protein variants with susceptibility to chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) is characterized by chronic airflow limitation and it is thought that neutrophils play a major role in the disease pathogenesis. Genetic polymorphism of the vitamin-D-binding protein (VDBP) gene is considered one of the candidates for variation in susceptibility to COPD. To evaluate the potential influences of VDBP gene polymorphisms on COPD, a case-control study was conducted in the Han population of north-east China. The VDBP polymorphic site was genotyped in 100 COPD patients and 100 controls. Genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism. A significantly higher proportion of VDBP-1F homozygosity was found in COPD patients, while the frequency of VDBP-2 homozygosity was significantly lower in COPD patients, which seemed to suggest that VDBP-2 homozygocity provided a protective effect. These data suggest that the VDBP gene may be involved in COPD susceptibility in Chinese Han population.

17 beta-estradiol affects proliferation and apoptosis of canine bone marrow mesenchymal stem cells in vitro.

Emerging researches in humans, pigs and mice, highlighted that estrogen plays a pivotal role in self-renewal and differentiation of bone marrow mesenchymal stem cells (BMSCs). The present study aimed at evaluating effects of 17 beta-estradiol (E2) on proliferation and apop-tosis of canine-derived bone marrow mesenchymal stem cells (cBMSCs) in vitro. The results showed that E2 supplementation at the concentration of 10-11 M promoted the proliferation of cBMSCs by CCK-8 assay and RT-qPCR analysis for the proliferation-related genes, with proliferating cell nuclear antigen (PCNA), cyclin-D1 (CCND1) being up-regulated and cyclin--dependent kinase inhibitor 1B (CDKN1B) being down-regulated. Contrarily, analysis of fluores-cence-activated cell sorting (FACS) and RT-qPCR demonstrated that E2 supplementation above 10-11 M had inhibitory effects on the proliferation of cBMSCs and induced apoptosis. Intriguingly,cBMSCs still possessed the capability to differentiate into osteoblasts and adipocytes with 10-11 M E2 addition. Taken together, this study determined the optimal culture condition of cBMSCs in vitro, and has important implications for further understanding the regulatory effect of E2 on the self-renewal of cBMSCs, which are helpful for the clinical application of BMSCs.

[Prospective cohort study on the risks of pre-pregnancy overweight, excessive gestational weight gain on macrosomia].

Objective: To investigate the risks of pre-pregnancy overweight, excessive gestational weight gain on macrosomia. Methods: We conducted one hospital-based cohort study, focusing on pregnant women from January 2015. All pregnant women attending to this hospital for maternal check-ups, were included in our cohort and followed to the time of delivery. Data related to general demographic characteristics, pregnancy and health status of those pregnant women, was collected and maternal pre-pregnant BMI and maternal weight gain were calculated. Logistic regression was used to explore the risk difference of pre-pregnancy BMI, excessive gestational weight gain on macrosomia. Results: The overall incidence of macrosomia in our cohort appeared as 6.6% (149/2 243). After adjusting the confounding factors including age and histories on pregnancy, pre-pregnancy overweight/obesity was associated with higher risks of macrosomia (OR=3.12, 95%CI: 1.35-7.22, P=0.008; OR=2.99, 95%CI: 1.17-7.63, P=0.022) when comparing to those with normal pre-pregnancy weight. Cesarean delivery and sex of the offspring were associated with higher risk of macrosomia, while excessive gestational weight gain showed no significant difference (OR=1.41, 95%CI: 0.96-2.09, P=0.084). Our data showed that Macrosomia was statistically associated with gestational weight gain (P=0.002). After controlling parameters as age, history of pregnancy and related complications of the pregnant women, results from the logistic regression showed that women with gestational inadequate weight gain having reduced risks to deliver macrosomia, when compared to those pregnant women with adequate weight gain (OR=0.52, 95%CI: 0.30-0.90, P=0.019). Conclusion: Pre-pregnancy overweight and obesity were on higher risks to macrosomia.

Intracarotid transplantation of bone marrow stromal cells increases axon-myelin remodeling after stroke.

The present study investigates the induction of axon and myelin remodeling as a possible mechanism by which treatment of stroke with bone marrow stromal cells improves neurological functional recovery. Adult male Wistar rats were subjected to 2 h of middle cerebral artery occlusion, followed by an injection of 2 x 10(6) rat bone marrow stromal cells or phosphate-buffered saline into the internal carotid artery 24 h later. Animals were killed at 28 days after stroke. Functional tests, histo- and immunohistochemical staining were performed. Significant functional recovery was found after bone marrow stromal cell administration in all the three tests performed (modified neurological severity score, adhesive-removal and corner tests). Bone marrow stromal cell treatment markedly increased vessel sprouting, synaptophysin expression and NG2 positive cell numbers and density in the cortical peri-infarct area. In bone marrow stromal cell-treated rats, the number of Ki-67 positive proliferating cells and oligodendrocyte precursor cells in the corpus callosum increased significantly in concert with the enhancement of the areas of the corpus callosum in both hemispheres. These results suggest that bone marrow stromal cells facilitate axonal sprouting and remyelination in the cortical ischemic boundary zone and corpus callosum, which may underlie neurological functional improvement caused by bone marrow stromal cell treatment.

Effect of the Fusarium toxins, zearalenone and deoxynivalenol, on the mouse brain.

The aim of this study was to find effects of Fusarium toxins on brain injury in mice. We evaluated the individual and combined effect of the Fusarium toxins zearalenone and deoxynivalenol on the mouse brain. We examined brain weight, protein, antioxidant indicators, and apoptosis. After 3 and 5days of treatment, increased levels of nitric oxide, total nitric oxide synthase, hydroxyl radical scavenging, and malondialdehyde were observed in the treatment groups. This was accompanied by reduced levels of brain protein, superoxide dismutase (apart from the low-dose zearalenone groups), glutathione, glutathione peroxidase activity, and percentage of apoptotic cells. By day 12, most of these indicators had returned to control group levels. The effects of zearalenone and deoxynivalenol were dose-dependent, and were synergistic in combination. Our results suggest that brain function is affected by zearalenone and deoxynivalenol.

The Fusarium toxin zearalenone and deoxynivalenol affect murine splenic antioxidant functions, interferon levels, and T-cell subsets.

This study aimed to evaluate the effects of the Fusarium toxin zearalenone (ZEA) and deoxynivalenol (DON) on splenic antioxidant functions, IFN levels, and T-cell subsets in mice. Herein, 360 mice were assigned to nine groups for a 12-day study. Mice were administered an intraperitoneal injection for 4 consecutive days with different concentrations of ZEA alone, DON alone, or ZEA+DON. Spleen and blood samples were collected on days 0, 3, 5, 8, and 12. Mice in each of the experimental groups showed dysreglated splenic antioxidant functions, IFN levels, and T-cell subset frequencies, suggesting that the immune system had been affected. The ZEA+DON-treated groups, especially the group that received a higher concentration of ZEA+DON (Group D2Z2), showed more obvious effects on the dysregulation of splenic antioxidant functions, IFN levels, and T-cell subsets. This finding suggested that DON and ZEA exerted synergistic effects.

Intratumoral injection of rhenium-188 microspheres into an animal model of hepatoma.

UNLABELLED: Intratumoral injection of 90Y microspheres is a potential alternative in the treatment of primary liver tumor. However, complicated preparation and lack of a gamma ray for imaging are the disadvantages of 90Y. In this study, we used 188Re, a generator-produced radioisotope with 155-keV gamma ray emission, to label microspheres. After intratumoral injection of 188Re microspheres into rats with hepatoma, biodistributions and survival times were analyzed. METHODS: Twelve male rats with hepatoma were killed at 1, 24 and 48 hr (4 rats at each time point) after intratumoral injection of approximately 7.4 MBq 188Re microspheres. Samples of various organs were obtained and used to calculate the tissue concentrations. In addition, 30 male rats bearing hepatoma were divided into two groups (15 rats in each group) to evaluate survival time. Group 1 received intratumoral injection of 37 MBq 188Re microspheres, whereas Group 2 served as the control group and received an intratumoral injection of 0.1 ml normal saline only. Survival time was calculated from the day of injection to 2 mo after treatment. RESULTS: Radioactivity in the tumor was very high throughout. Biological half-time was 170.8 hr. Radioactivity in the lung was 1.78% injected dose (i.d.)/g at 1 hr but declined rapidly over time. The concentration in the urine was approximately 6.14% i.d./ml after the first hour and rapidly declined thereafter. The concentrations of radioactivity in other organs, such as normal liver, muscle, spleen, bone, testis and whole blood, were quite low throughout the study. Twelve of 15 (80%) of rats survived over 60 days after intratumoral injection of 188Re microspheres, whereas only 4 of 15 (26.7%) survived more than 60 days after injection of normal saline only. The difference between the groups was significant (p < 0.05). CONCLUSION: Rhenium-188 offers cost-effectiveness, on-site availability, short half-life, energetic beta particle, emission of gamma photons for imaging, easy preparation, easy clinical administration and apparent lack of radiation leakage from the treated tumor. Direct intratumoral injection of 188Re microspheres is extremely attractive as a clinical therapeutic alternative in hepatoma patients.

Low prevalence of human papillomavirus infection in esophageal squamous cell carcinomas from North America: analysis by a highly sensitive and specific polymerase chain reaction-based approach.

Several studies have documented the frequent occurrence of human papillomavirus (HPV) DNA in esophageal squamous cell carcinomas (ESCC) in patients from geographic regions where the incidence of this type of cancer is high, such as parts of China. However, the prevalence of HPV infection in ESCC in patients from low incidence geographic regions, such as North America, remains controversial. Therefore, this study evaluates the prevalence of HPV in ESCC in patients from North America, a region where the population is considered at low risk for the development of this neoplasm. ESCCs in 51 patients from three North American cities were analyzed for the presence of HPV DNA by a highly sensitive and specific polymerase chain reaction (PCR) method. Tumor DNA was extracted from formalin-fixed, paraffin-embedded tissue specimens and assayed by PCR using an L1 HPV consensus sequence primer, as well as HPV 16 and HPV 18 E7 region primers. The use of consensus primers to the L1 region allows for detection of most known HPV types and many novel HPV types. Appropriately sized reaction products were analyzed by restriction fragment length polymorphism (RFLP) to confirm the presence and type of HPV, and to exclude products produced by amplification of human DNA sequences. After complete analysis, only one case (2%) of ESCC was HPV DNA positive. This case was independently confirmed using L1 and E7 consensus primers as HPV type 16 and was the only case that tested positive with either assay. These results show that, in contrast to geographic regions where ESCC is prevalent, HPV infection occurs infrequently in association with ESCC in patients from North America.

Disparities in mean age and histopathologic grade between human papillomavirus type-specific early cervical neoplasms.

Noninvasive squamous and glandular precursor lesions associated with human papillomavirus (HPV) types 16 and 18 have been reported to vary in morphology. HPV 16 is associated predominantly with high-grade squamous intraepithelial lesions (HSIL; cervical intraepithelial neoplasia (CIN 2 and 3), and HPV 18 is associated with low-grade squamous intraepithelial lesions (condyloma/CIN 1) and CIN 3/adenocarcinoma in situ (ACIS). This study explored the relationship of morphologic growth pattern in these precursor groups with age of presentation. One hundred fourteen CIN lesions (including those with ACIS), associated with HPV 16 or 18, were subdivided into well-differentiated low- and high-grade SIL (CIN 1 and 2, respectively), poorly differentiated HSIL (CIN 3) with or without ACIS. HPV was detected by polymerase chain reaction (PCR) amplification with L1 consensus or type-specific E7 primers and typed by restriction fragment length polymorphism (RFLP) analysis. Age of the patient was obtained from the pathology report. Mean age for each group was as follows: Low-risk HPVs, 25 years; HPV 18 CIN 1-2, 21.6 yrs; HPV 18 CIN 3/ACIS, 35.2 yrs; HPV 16 CIN 1,2, 25.9 yrs; and HPV 16 CIN 3, 29.8 yrs. There were significant differences in mean ages between HPV 18 CIN 1 and 2 and HPV 16 CIN 1 to 2 (P = .04), HPV 16 CIN 1-2 and CIN 3 (P = .01) and HPV 18 CIN 1 to 2 and HPV 18 CIN 3/ACIS (P = .00001). None of the cases of HPV 18-associated CIN3/ACIS was associated with a CINI lesion. The disparity in mean ages between well and poorly differentiated HPV 16/18 related that precursor lesions could reflect factors such as morphologic progression with increasing age, different rates of lesion persistence, depending on grade, or efficiency of detection between the two groups. The marked difference in mean age between HPV 18-associated CIN 1-2 and CIN 3/ACIS, combined with their lack of coexistence in the same cervix, raises alternate possibilities that specific viral or host factors may determine the morphological phenotype associated with some HPV 18 infections. In the latter, the possibility that age independently confers an increased risk for higher-grade lesions should be considered.

Prevalence and histologic significance of cervical human papillomavirus DNA detected in women at low and high risk for cervical neoplasia.

OBJECTIVES: To determine and compare the prevalence and histologic significance of human papillomavirus (HPV) nucleic acids in cervical specimens from women at low (routine hysterectomy) and high (suspicion of cervical neoplasia) risk for cervical neoplasia. METHODS: Cervical brushings were taken from the cervices of hysterectomy and conization or loop electrical excision specimens and analyzed for HPV nucleic acids by polymerase chain reaction and restriction fragment length polymorphism analysis. Histopathology was confirmed by review of reports or, for HPV-positive results, re-review of the histopathology. Statistical analysis used Student t test or Fisher exact test. RESULTS: Four hundred seventeen and 43 low- and high-risk cervices, respectively, were studied. Statistically significant differences were observed in the index of HPV positivity between the low- and high-risk groups (1.7 versus 42%, P < .001) and the proportion of HPV being cancer-associated HPV types (14 versus 78%, P = .005). None of the 417 cervices from low-risk women contained HPV 16. In the high-risk group, histologically confirmed cervical intraepithelial neoplasia lesion was statistically more likely to be associated with HPV (59 versus 13%, P = .005). CONCLUSION: Cervices from routine, low-risk hysterectomies in predominately middle-aged women have an extremely low index of cancer-associated HPVs. Considering the strong association of HPV with histologically proven disease, prospective studies exploring the relationship of cancer-associated HPVs to neoplasia in middle-aged women merit consideration.

Histologic study of effects of radiation synovectomy with Rhenium-188 microsphere.

Rhenium-188 microsphere is a relatively new radiation synovectomy agent developed for the treatment of rheumatoid arthritis. It has been shown that the levels of unwanted extra-articular radiation are negligible with this agent. A histologic study was conducted to assess the effect of radiation synovectomy on synovium and articular cartilage after intra-articular injection of various doses of Re-188 microspheres into the knee joints of rabbits. Intra-articular injection of Re-188 microspheres into rabbit knee joints resulted in mild reactive inflammation and thrombotic occlusion of vessels which subsided rapidly. Sclerosis of subsynovium could be seen 12 weeks after injection. No evidence of damage to articular cartilage was noted. There was no significant difference in the articular pattern after injection of 0.3 or 0.6 mCi Re-188 microspheres. This study suggests that a treatment dose of Re-188 microspheres causes transient inflammation of synovium without any detectable damage to the articular cartilage of knee joint.

Pentavalent Tc-99m dimercaptosuccinic acid imaging of hepatocellular carcinoma.

Tc-99m (V)-dimercaptosuccinic acid (DMSA) has been used to image various kinds of tumors. However, from a review of the literature, it has never been applied to hepatocellular carcinoma (HCC). Low uptake of Tc-99m (V)-DMSA has been demonstrated in normal liver tissue, thus, Tc-99m (V)-DMSA may be useful for the detection of HCC. Nine male patients with focal nodular HCC were studied with sequential X-ray CT, Tc-99m (V)-DMSA and Tc-99m phytate liver scan. Our data showed that eight patients had increased uptake of Tc-99m (V) DMSA in HCC. Four cases demonstrated higher Tc-99m (V) DMSA uptake in HCC than in adjacent liver, and four cases demonstrated HCC uptake equal to liver uptake. One case showed no uptake of Tc-99m (V) DMSA in HCC. The detection sensitivity of Tc-99m (V)-DMSA was 88.9%. From our early results, Tc-99m (V)-DMSA is a readily available tumor imaging agent that appears to accumulate in HCC.