Stability of IgE in Postmortem Plasma and Hemolyzed Samples.

ABSTRACT: Objective To investigate the stability of IgE in postmortem plasma and hemolyzed samples under different storage conditions and freezing-thawing. Methods Thirty nine cardiac blood samples were collected from non-frozen corpses with the postmortem interval of less than 48 hours, including 20 plasma samples and 19 hemolyzed samples taken from whole blood. The samples were stored at -20 ℃, 4 ℃ and 25 ℃ for 28 d and at -80 ℃ for 1 year to evaluate the stability of IgE under different storage conditions. Repeated freezing-thawing treatment was conducted for 5 times to explore the stability of IgE in postmortem plasma and hemolyzed samples. IgE concentration in plasma and hemolyzed samples was detected by electroluminescence before and after treatment. Results The degradation rates of IgE in plasma samples under the three storage conditions, -20 ℃, 4 ℃ and 25 ℃ were close. After 28 d, the mean value was about 15%, the degradation speed of IgE in hemolyzed samples was faster than that of plasma under the same condition （P<0.05） and the degradation rate was faster than other two conditions under 25 ℃ （P<0.05）. The differences in the concentration of plasma samples after freezing at -80 ℃ for 1 year and that before freezing had no statistical significance （ P>0.05）, while the concentration of hemolyzed samples was degraded after freezing at -80 ℃ for 1 year （P<0.05）. The differences between the detection results of plasma and hemolyzed samples after repeated freezing-thawing for 5 times and that before freezing-thawing showed no statistical significance （ P>0.05）. Conclusion IgE has good freezing-thawing stability in postmortem plasma and hemolyzed samples. Stability of IgE is better in postmortem plasma samples than hemolyzed samples, thus it is recommended to separate plasma from postmortem blood samples as soon as possible in forensic practice.

Application of Hollow Fiber Ultrafiltration Technology in the Detection of Total IgE of Hemolytic Blood after Death.

ABSTRACT: Objective To investigate the treatment effect of hollow fiber ultrafiltration technology on hemolytic samples and the differences between IgE concentration and serum concentration before hemolysis in ultrafiltrate. Methods The 33 postmortem blood samples of non-frozen corpses within 72 hours after death were collected, 4 mL blood was taken from each case, among which 1 mL was centrifuged to get serum, and the remaining 3 mL blood was frozen-thawed 3-5 times to cause complete hemolysis. The 2 mL hemolytic samples were processed by hollow fiber ultrafiltration to obtain ultrafiltrate. The hemoglobin concentration in serum, complete hemolytic sample and ultrafiltrate was determined by Van-Zij solution-cyanated methemoglobin assay method, and the total IgE in serum and ultrafiltrate was determined by electrochemical luminescence method. Results The hemoglobin concentration in ultrafiltrate was significantly lower than that in complete hemolytic samples （P<0.05）. There was a good correlation between the total IgE detection values of ultrafiltrate and serum （r=0.984）. The difference between the serum and the value of IgE in ultrafiltrate after correction had no statistical significance, and the differences between the two in positive rates had no statistical significance （P>0.05）. Conclusion Ultrafiltration technology has a good treatment effect on complete hemolytic samples, and the correction value of ultrafiltrate detection is close to the serum level before hemolysis, and therefore, it can be applied to the detection of total IgE of frozen corpse hemolytic samples.

[Effects of artesunate combined with bortezomib on apoptosis and autophagy of acute myeloid leukemia cells in vitro and its mechanism].

Objective: To investigate the effects of artesunate combined with bortezomib on the proliferation, apoptosis and autophagy of human acute myeloid leukemia cell lines MV4-11, and its mechanisms. Methods: MTT method was used to determine the anti-proliferation effect of different concentrations of artesunate, bortezomib and their combination on MV4-11 cells. The cell apoptosis were analyzed by flow cytometry. The expression of cleaved-Caspase-3, Bcl-2 family protein (Bcl-2, Mcl-1, Bim, Bax) and autophagy-related protein LC3B were assayed by Western blot. Results: Artesunate displayed a proliferation inhibition effect on MV4-11 with dose- and time-dependent manner, the IC(50) of artesunate on MV4-11 after 48 hours was 1.44 µg/ml. Bortezomib displayed a proliferation inhibition effect on MV4-11 with dose-dependent manner, the IC(50) of bortezomib on MV4-11 after 48 hours was 8.97 nmol/L. The combination of artesunate (0.75, 1.0 µg/ml) and Bortezomib (6, 8 nmol/L) showed higher inhibition on MV4-11 than artesunate or bortezomib alone in the same concentration gradient after 48 hours (P<0.05) . The cooperation index of the two drugs were all less than 1. The 48 h apoptotic rate of artesunate (1.5 µg/ml) on MV4-11 was (15.27±2.18) %, (19.85±3.23) % of bortezomib (8 nmol/L) , (81.67±5.96) % of combination of the two drugs, significantly higher than the single group (P<0.05) . When combination of the two drugs on MV4-11 after 24 hours, the levels of pro-apoptotic protein Bim and the cleaved activation of Caspase-3 and autophagy-related protein LC3B were up-regulated and the anti-apoptotic protein Bcl-2 expressions was down-regulated. Conclusion: Combination of artesunate with bortezomib shows a significant synergistic effects on proliferation, apoptosis and autophagy of MV4-11 cell lines, which may be associated with Bcl-2 family proteins expression.

Effect of Corpse Cryopreservation on Forensic Pathological Identification.

Because cryopreservation can effectively slow down the putrefaction and prolong the preservation time of corpses, it has become the main way of corpse preservation in China. However, it may cause a certain degree of non-specific effects on the corpses and thus interfere with forensic pathological identification. This paper summarizes relevant problems reported in domestic and foreign literature reports and practical identification, and analyzes the effects of cryopreservation on corpses from aspects of anatomical findings, histomorphology, postmortem biochemistry, and postmortem imaging, therefore to provide Chinese forensic workers assistance on problems in their practice of forensic pathology.

Statistical Analysis of the Heart and Lung Mass in Forensic Anatomical Cases and Its Forensic Significance.

ABSTRACT: Objective To investigate the differences of heart mass and lung mass in forensic autopsy cases and to explore their application value in forensic identification. Methods The data from 1 614 autopsy cases accepted by center of Medico-legal Investigation of China Medical University between 2007 to 2016 were collected. The correlation of heart and lung mass with age, height, body weight, and body mass index （BMI）, as well as differences in different causes of death were analyzed by GraphPad Prism 6.0 software. Results The heart mass and lung mass of males were higher than those of females （P<0.05）. The heart mass of males and females was positively correlated with age, height, body weight and body mass index （BMI） （P<0.05）. The heart mass of patients dying from sudden cardiac death （SCD） was significantly higher than those dying from other causes of death. The lung mass of patients dying from drowning and delayed treatment was higher than those dying from other causes of death （P<0.05）. Conclusion In forensic practice, measurement of the heart mass and lung mass has certain significance for differential diagnosis and diagnosis of different causes of death.

Research Progress and Forensic Identification of Alcoholic Cardiomyopathy.

ABSTRACT: With the rapid development of the social economy in China, the incidence of diseases caused by excessive drinking is gradually increasing as well. Alcoholic cardiomyopathy refers to long-term high intake of ethanol, and has typical dilated cardiomyopathy characteristics, such as, hemodynamic changes, symptoms, signs, and morphological features. It is a kind of cardiomyopathy that excludes other causes of dilated cardiomyopathy. Due to the lack of specific pathological changes, the forensic pathological identification of alcoholic cardiomyopathy can only be based on the patient's medical history and by ruling out other causes of cardiomyopathy. This paper reviews the pathogenesis and forensic identification of alcoholic cardiomyopathy in order to provide reference for forensic pathologists and clinicians.

Forensic Pathological Identification and Antidiastole of Commotio Cordis.

Commotio cordis （CC） is the acute death caused by the cardiac rhythm disorder after a sudden blunt external force to the precordium of a healthy person without previous heart disease. As one type of violent heart damage, CC is rare with relatively small external force and sudden death, therefore causing disputes. This paper reviews the epidemiology, mechanisms and the key points in forensic identification of CC, discusses the identification and antidiastole of CC, myocardial contusion, sudden cardiac death and death from inhibition, and provides assistance to forensic pathologists to identify such causes of death.

Correlation of acetabular chondrocyte apoptosis with caspase-3 and Bcl-2 expression in developmental dislocations of the hip.

This study aimed to determine whether abnormal apoptosis is present in acetabular cartilage in early developmental dislocations of the hip (DDH), and if so, whether it is correlated with the expression of caspase-3 and Bcl-2. DDH was induced in 24 4-week-old New Zealand white rabbits. Acetabular cartilage specimens from the experimental and control groups were stained with hematoxylin and eosin (H&E). Animals from the experimental group developed acetabular dysplasia. Apoptotic chondrocytes were observed by ultrastructural electron microscopy and H&E. TUNEL assays revealed significantly greater acetabular chondrocyte apoptosis in the treated samples as compared to the control. Significantly higher caspase-3 expression and lower Bcl-2 expression were also measured in the DDH group compared with the control. We conclude that excessive apoptosis does occur in acetabular cartilage with DDH, and is positively correlated with high caspase-3 expression as well as low Bcl-2 expression.

[Efficacy of GLIDE chemotherapy for patients with newly diagnosed advanced-stage or relapsed/refractory extranodal natural killer cell lymphoma].

Objective: To evaluate the efficacy of gemcitabine, asparaginase , ifosfamide, dexamethasone and etoposide (GLIDE) combination for patients with newly diagnosed advanced-stage or relapsed/refractory extranodal natural killer cell lymphoma (ENKL). Methods: Fourty-two newly diagnosed advanced-stage or relapsed/refractory ENKL were enrolled from March 2010 to March 2016. Patients were treated with GLIDE for median 3 (2-6) cycles. Complete response (CR) rate, early CR (after 2 cycles) rate were evaluated after all treatment finished. Progression free survival (PFS) and overall survival (OS) were analyzed by Kaplan-Meier method and multivariate by Cox proportional hazards models. Results: Thirty-one (73.8%) patients achieved CR with 22 (52.4%) in early CR after 2 cycles of GLIDE, and 14 underwent autologous stem cell transplantation (ASCT) after achieved CR. One year PFS and OS were 65.6% and 82.7%, 4 year PFS and OS were 48.2% and 63.1%, respectively, with a median PFS of 30.5 months. Multivariate analysis indicated ECOG score 0-1 and ASCT after CR were independent prognostic factors for less relapse and longer survival. Conclusion: GLIDE is an effective regiment for newly diagnosed advanced-stage and relapsed/refractory ENKL.

The complete mitochondrial genome of Pseudanoplocephala crawfordi and a comparison with closely related cestode species.

Pseudanoplocephala crawfordi is an important zoonotic cestode of economic significance and public health concern. In spite of its significance as a pathogen, the systematics, genetics, epidemiology and biology of this parasite remain poorly understood. In the present study, we sequenced and characterized the complete mitochondrial (mt) genome of P. crawfordi, which is 14,192 bp long and encodes 36 genes, including 12 protein-coding genes, 22 transfer RNA genes and two ribosomal RNA genes. Phylogenetic analysis of the concatenated amino acid sequences using the Bayesian inference (BI) method showed that P. crawfordi was closely related to the family Hymenolepididae. Considering that the taxonomic status of P. crawfordi has been controversial when based only on morphological features, the mt genome obtained here will provide novel molecular markers to ascertain the phylogenetic position of this parasite accurately.

Molecular characterization of Cryptosporidium spp. in pre-weaned calves in Shaanxi Province, north-western China.

Cryptosporidium, a worldwide protozoan parasite, is one of the most common causes of diarrhoea in humans and animals. The aim of the present study was to determine Cryptosporidium species/genotypes in pre-weaned calves in Shaanxi Province using PCR and sequencing based on the small subunit rRNA gene. A total of 258 faecal samples were collected from pre-weaned calves in 19 different farms from six areas in Shaanxi Province, north-western China. Cryptosporidium infection was detected in 14 of 19 farms (73.7 %), with a total prevalence of 20.2 % (52/258). Both dairy and Qinchuan (beef) cattle were found with Cryptosporidium infection. Three Cryptosporidium species, namely Cryptosporidium bovis (n = 26), Cryptosporidium andersoni (n = 14) and Cryptosporidium ryanae (n = 12), were detected in pre-weaned calves in Shaanxi Province, with C. bovis (in 12 farms) identified as the most common species on cattle farms. Two additional and previously unknown C. ryanae genotypes, CRTypes III and IV, were observed in the present study. However, the zoonotic species, Cryptosporidium parvum, was not detected in this study, which suggested a low zoonotic potential in Cryptosporidium-infected pre-weaned calves in this province.

Characterization of Dicrocoelium dendriticum isolates from small ruminants in Shaanxi Province, north-western China, using internal transcribed spacers of nuclear ribosomal DNA.

The genetic variations in internal transcribed spacers (ITS) spanning ITS-1, 5.8S and ITS-2 rDNA of Dicrocoelium dendriticum, isolated from sheep and goats in four geographical regions in Shaanxi province, were examined. The lengths of ITS-1, 5.8S and ITS-2 rDNA sequences for D. dendriticum were 749 bp, 161 bp and 234 bp, respectively. Intra-specific sequence variations of D. dendriticum were 0-0.5% for ITS-1 and 0-1.3% for ITS-2 rDNA, while the inter-specific variations among species in genus Dicrocoelium in ITS-2 rDNA were 3.4-12.3%. Phylogenetic analysis based on sequences of ITS-2 rDNA showed that all D. dendriticum isolates in the present study were grouped with reference D. dendriticum isolates from sheep and goats, and D. dendriticum isolates from cattle and Japanese serow were clustered in a sister clade. However, the phylogenetic tree could not reveal geographically genetic relationships of D. dendriticum isolates in different origins and hosts. These findings provided basic information for further study of molecular epidemiology and control of D. dendriticum infection in Shaanxi province as well as in the world.

Molecular approaches to differentiate three species of Nematodirus in sheep and goats from China based on internal transcribed spacer rDNA sequences.

Internal transcribed spacer (ITS) rDNA sequences of three Nematodirus species from naturally infected goats or sheep in two endemic provinces of China were analysed to establish an effective molecular approach to differentiate Nematodirus species in small ruminants. The respective intra-specific genetic variations in ITS1 and ITS2 rDNA regions were 0.3-1.8% and 0-0.4% in N. spathiger, 0-6.5% and 0-5.4% in N. helvetianus, and 0-4.4% and 0-6.1% in N. oiratianus from China. The respective intra-specific variations of ITS1 and ITS2 were 1.8-4.4% and 1.6-6.1% between N. oiratianus isolates from China and Iran, 5.7-7.1% and 6.3-8.3% between N. helvetianus samples from China and America. For N. spathiger, compared with samples from China, sequence differences in ITS1 rDNA were 0.3-2.4% in isolates from America, 0.3-2.9% in New Zealand and 2.1-2.4% in Australia. Genetic variations in ITS2 rDNA of N. spathiger were 0-0.4% between samples from China and America, and 0-0.8% between samples from China and New Zealand. Using mutation sites, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and specific PCR techniques were developed to differentiate these three Nematodirus species. The specific PCR assay allowed the accurate identification of N. oiratianus from other common nematodes with a sensitivity of 0.69 pg and further examination of Nematodirus samples demonstrated the reliability of these two molecular methods.

Characterization of Oesophagostomum asperum and O. columbianum by internal transcribed spacers of nuclear ribosomal DNA.

In the present study, the internal transcribed spacers (ITS) of ribosomal DNA (rDNA) of Oesophagostomum asperum and O. columbianum were amplified and sequenced. The ITS-1, 5.8S and ITS-2 rDNA sequences of O. asperum were 374 bp, 153 bp and 259 bp in length, respectively, and the corresponding sequences of O. columbianum were 259, 153 and 218 bp in length, respectively. Sequence differences in the ITS-1 and ITS-2 rDNA between the two Oesophagostomum species were 9.5-10.2% and 12.7-13.9%, respectively. Sequence differences in the ITS-1 and ITS-2 rDNA among members of the genus Oesophagostomum were 2.5-11.6% and 6.8-22.3%, respectively. Based on genetic markers in the ITS rDNA, an effective polymerase chain reaction (PCR) approach was developed to differentiate O. columbianum from O. asperum with a sensitivity of 0.2 ng/µl DNA. Since accurate characterization of parasites at different taxonomic levels is essential for population genetic studies and control of parasitosis, the present findings have important implications for studying epidemiology, taxonomy and population biology, as well as for the control of oesophagostomiasis.

Potential role of proteomics in the diagnosis of lymphoma: a meta-analysis.

Surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF-MS) has been approved for identifying biomarkers and diagnosing many diseases such as lymphomas. It is arguable whether the SELDI technique has its value of diagnostic accuracy for lymphomas. The purpose of our study is to determine the diagnostic accuracy of SELDI-TOF-MS for diagnosing lymphomas. The Cochrane Central Register of Controlled Trials, MEDLINE, Pub Med, EMBASE, the Chinese Biomedical Literature Database, the China Academic Journals Full-text Database, and the Chinese Scientific Journals Database were searched systematically for potential studies. Reference lists of included studies and review articles were also reviewed. All studies that reported data on patients with a confirmed diagnosis of lymphomas and that compared the measurement of SELDI-TOF-MS with pathology standard were considered for inclusion. Eleven studies were included in the systematic review. The ranges of the diagnostic value of SELDI-TOF-MS for lymphoma were as follows: sensitivity (SEN) was 0.69-0.96; specificity (SPE) was 0.70-1.00; positive likelihood ratio (PLR) was 2.99-96.09; negative likelihood ratio (NLR) was 0.04-0.35; and diagnostic odds ratio (DOR) was 18.13-1250.71, respectively. Further, we analysed serum samples as a subgroup, and the pooled endpoints were as follows: pooled SEN was 0.89 (0.85-0.91); pooled SPE was 0.91 (0.88-0.93); pooled PLR was 12.35 (5.36-28.44); pooled NLR was 0.13 (0.09-0.20); and pooled DOR was 101.04 (39.57-258.04), respectively. SELDI-TOF-MS showed high accuracy in identifying lymphoma and could be a useful screening tool for diagnosing lymphoma patients.

Anti-angiogenesis therapy based on the bone marrow-derived stromal cells genetically engineered to express sFlt-1 in mouse tumor model.

BACKGROUND: Bone marrow-derived stromal cells (BMSCs) are important for development, tissue cell replenishment, and wound healing in physiological and pathological conditions. BMSCs were found to preferably reach sites undergoing the process of cell proliferation, such as wound and tumor, suggesting that BMSCs may be used as a vehicle for gene therapy of tumor. METHODS: Mouse BMSCs were loaded with recombinant adenoviruses which express soluble Vascular Endothelial Growth Factor Receptor-1 (sFlt-1). The anti-angiogenesis of sFlt-1 in BMSCs was determined using endothelial cells proliferation inhibition assay and alginate encapsulation assay. The anti-tumor effects of BMSCs expressing sFlt-1 through tail-vein infusion were evaluated in two mouse tumor metastases models. RESULTS: BMSCs genetically modified with Adv-GFP-sFlt-1 could effectively express and secret sFlt-1. BMSCs loaded with sFlt-1 gene could preferentially home to tumor loci and decrease lung metastases and prolong lifespan in mouse tumor model through inducing anti-angiogenesis and apoptosis in tumors. CONCLUSION: We demonstrated that BMSCs might be employed as a promising vehicle for tumor gene therapy which can effectively not only improve the concentration of anticancer therapeutics in tumors, but also modify the tumor microenvironment.

Synthesis and characterization of aspartic acid complexes of antimony and bismuth triiodide.

New bioinorganic complexes of the aspartic acid with the antimony or bismuth triiodide were synthesized by a direct solid-solid reaction at room temperature. The formula of the complex is MI(3)[OOCCH(2)CH(NH(2))CO](2.5) x 2.5H(2)O (M = Sb, Bi). The complex may be a dimer with bridge structure. The crystal structure of the complexes belongs to a triclinic system. The lattice parameters are a = 0.9883 nm, b = 1.4284 nm, c = 2.0114 nm, alpha = 94.46 degrees , beta = 99.76 degrees and gamma = 100.1 degrees for the complex of antimony and a = 0.9756 nm, b = 1.4560 nm, c = 1.9875 nm, alpha = 94.18 degrees , beta = 97.25 degrees and gamma = 101.16 degrees for the complex of bismuth. The infrared spectra and thermal analyses can demonstrate the complex formation between the aspartic acid and the antimony or bismuth ion.

Preparation of new amino acid complex nanoparticles of bismuth and leucine.

The new amino acid complex nanoparticles of bismuth and leucine can be prepared very easily by a room temperature solid-solid reaction. The various characterizations indicate that the formula of the complex is BiCl[(CH(3))(2)CHCH(2)CHNH(2)COO](2)1.5H(2)O. The crystal structure of the solid complex belongs to monoclinic system with the lattice parameters: a = 1.6036 nm, b = 1.9903 nm, c = 2.1979 nm and beta=108.06 degrees . The new solid complex is nanoparticles with average size about 80 nm.

[Diagnostic study of pulmonary mass under ultrasonography guided aspiration biopsy by four types of needle].

One hundred and thirty patients with pulmonary mass were studied under ultrasonography guided aspiration biopsy by four types of needle. The cytological positive cases were 57.1% in fine needle (22G), 79.6% in large needle (18G), 90.9% in canaliculated needle (18G) and 80.9% in cutting needle (14G), and the pathological positive cases were 0, 64.4%, 86.3% and 92.8% respectively. These data showed that the positive rate of fine needle puncture was the lowest one. Based on the pathological finding, canaliculated and cutting needle seem to be better than large one and it is the best choice to perform the pulmonary aspiration biopsy.

Mössbauer effect studies on some bioinorganic complexes of europium.

The bioinorganic complexes of europium with N-acetyl-DL-alanine, N-acetyl-DL-valine, and DL-alanyl-DL-alanine have been synthesized and the Mössbauer spectra at room temperature have been measured for these solid state complexes. The Mössbauer parameters indicate that the water molecules in these complexes are not directly linked to the central europium ion and are outside the coordination sphere of europium and biological ligands, and that the chemical bond between the europium ion and the ligands may be predominantly ionic in character, with the possibility of partial covalent contribution.