A gross anatomical study of the styloid process of the temporal bone in Japanese cadavers.

BACKGROUND: The incidence of an elongated styloid process (SP) and average length and diameter of SP have not been reported using Japanese cadavers. Data on the female-to-male ratio of an elongated SP vary. We calculated the average length and diameter of SP in Japanese cadavers and compared SP lengths between sexes. MATERIALS AND METHODS: Twenty-seven sides (right and left of bodies) in males and 51 sides in females were analysed. Measurements were obtained from the inferior external acoustic meatus to the distal tip of the SP. SP diameters at the proximal base, midpoint, and distal tip were measured. SP > 30 mm was considered elongated. We used Welch's t-test for the statistical analysis. Fisher's exact two-tailed test was also performed to analyse the female-to-male elongation ratio. A p-value < 0.05 was considered statistically significant. RESULTS: Styloid process elongation prevalence was 29.5% in our sample. The average full length was 27.04 ± 7.88 mm overall; the average diameters were 5.41 ± 1.77 mm at the proximal base and 2.21 ± 1.22 mm at the distal tip. The average SP measurement was 26.81 ± 5.92 mm in males and 27.16 ± 8.79 mm in females (p = 0.74). The female-to-male ratio of SP elongation was 1:2 (p = 0.041). Females had longer full lengths of non-elongated SPs than males (p = 0.004). Males had wider diameters at the proximal base of elongated SPs than females (p = 0.017). CONCLUSIONS: The average length of SP was 27.04 mm in the Japanese population and about 30% of the Japanese presented SP ≥ 30 mm. Male had significantly higher rate than female among the SP ≥ 30 mm, and female had significantly longer SPs than male among the SP < 30 mm. Anatomically, the SP gets narrow as distally goes. Our anatomical findings would be beneficial to creating treatment plans, diagnosis, and surgery.

Variations in the gonadal artery with a single common trunk: embryological hypotheses by observation.

BACKGROUND: A gonadal artery originates as a branch of the abdominal aorta and renal artery inferior to the level of origin of the renal arteries. Variations in multiple right testicular arteries (RTAs) arising from the abdominal aorta are common. We aimed to re-evaluate the unusual courses of gonadal arteries with a single common trunk in relation to the inferior vena cava and left renal vein and explain the developmental anatomy. MATERIALS AND METHODS: The observational cross-sectional study was performed on 54 Japanese adult cadavers (29 men and 25 women). We examined the literature and developed embryological hypotheses on the single common trunk of the gonadal artery. RESULTS: The gonadal artery, testicular artery, and ovarian artery arose from the abdominal aorta in 93.1%, 96.3%, and 89.6% of cases, respectively, and from the renal artery in 4.9%, 3.7%, and 6.3% of cases, respectively. We found two rare variations in the RTAs observed during the routine dissection of two male cadavers; in these two cases, a single common trunk of the RTAs originated from the abdominal aorta. A single common trunk was found in 3.7% of cadavers, 2.0% of sides, and 2.0% of arteries in the gonadal artery and in 6.9% of cadavers, 3.8% of sides, and 3.7% of arteries in the testicular artery. All cases of the single common trunk, including those in past reports, were observed only in men. CONCLUSIONS: Knowledge of the variations in RTAs has important clinical consequences for invasive and non-invasive arterial procedures. In addition, this variation provides a new interpretation of the embryology of the gonadal artery. Variations similar to our findings have not been previously reported. Therefore, different variations concerning the RTA should be considered during surgical and non-surgical evaluations.

MiR-424 suppressed viability and invasion by targeting to the DCLK1 in neuroblastoma.

OBJECTIVE: Neuroblastoma is the most frequent tumor of sympathetic nervous system in infants. MiRNAs acted as oncogenes or tumor suppressors in the process of tumor development. We aim at exploring the functions of miRNA in neuroblastoma. PATIENTS AND METHODS: Cell viability and invasion were evaluated by Cell Counting Kit-8 (CCK-8) and transwell assays. Western blot was utilized to assess the protein expression associated with epithelial-mesenchymal transition (EMT) markers. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to calculate the mRNA levels of miRNA and gene. RESULTS: MiR-424 was downregulated while doublecortin like kinase 1 (DCLK1) was upregulated in neuroblastoma tissues and cells compared to adjacent non-tumor and normal spongiocyte cells. MiR-424 suppressed cell viability, invasion, and EMT by targeting DCLK1. MiR-424 regulated the expression of DCLK1 by directly binding to the 3'-untranslated region (UTR) of DCLK1 mRNA in SK-N-SH and Be2C cells. DCLK1 reversed partial functions of miR-424 in neuroblastoma. CONCLUSIONS: MiR-424 suppressed cell viability, invasion, and EMT by directly targeting the 3'-UTR of DCLK1 mRNA. The newly identified miR-424/DCLK1 axis provides novel insights into the pathogenesis of neuroblastoma.

Microvesicles containing microRNA-216a secreted by tubular epithelial cells participate in renal interstitial fibrosis through activating PTEN/AKT pathway.

OBJECTIVE: The aim of this study was to elucidate the role of microRNA-216a in microvesicles (MVs) during the process of renal interstitial fibrosis and to investigate its underlying mechanism. MATERIALS AND METHODS: Unilateral ureteral occlusion (UUO) model was first established in mice, and kidney tissues and urine in the obscured kidney were collected. NRK-52E cells were induced with 5 ng/mL transforming growth factor-ß1 (TGF-ß1) for constructing the renal interstitial fibrosis model in vitro. Subsequently, the expression levels of E-cadherin, α-smooth muscle actin (α-SMA) and fibronectin (FN) in NRK-52E cells induced with or without TGF-ß1 were determined, respectively. The culture medium was collected from NRK-52E cells of the control group (without TGF-ß1 induction) and the TGF-ß1 group (TGF-ß1 induction), and MVs were observed. Afterward, NRK-52E cells were treated with MVs isolated from the control group or the TGF-ß1 group, followed by detecting the expressions of E-cadherin, α-SMA and FN. Meanwhile, the expression levels of CD63, microRNA-216a, PTEN and p-AKT were determined as well. The microRNA-216 level in kidney tissues and urine of UUO mice were determined. Furthermore, the expressions of PTEN and p-AKT in mouse kidney tissues were accessed by Western blot and quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). RESULTS: TGF-ß1 induction in NRK-52E cells gradually downregulated E-cadherin, whereas upregulated α-SMA and FN with the prolongation of induction time. MVs isolated from the culture medium of the TGF-ß1 group downregulated E-cadherin, and upregulated FN and α-SMA. The expression levels of CD63 and microRNA-216a were markedly higher in the TGF-ß1 group compared with the control group. Downregulated PTEN and upregulated p-AKT were observed in TGF-ß1-induced cells at both mRNA and protein levels. Besides, microRNA-216a expression in mouse kidney tissues and urine from obscured kidney was remarkably increased with the prolongation of UUO. Consistent with those in NRK-52E cells, the protein level of PTEN was significantly decreased, whereas p-AKT was markedly increased with the prolongation of UUO. CONCLUSIONS: MVs containing microRNA-216a secreted by injured proximal tubular epithelial cells participate in renal interstitial fibrosis by activating the PTEN/AKT pathway.

Meta-analysis of the effects of helmet-assisted non-invasive ventilation in the treatment of acute respiratory failure.

OBJECTIVE: To study the efficacy of helmet-assisted non-invasive ventilation and conventional ventilation in the treatment of acute respiratory failure (ARF). MATERIALS AND METHODS: Cochrane Library, PubMed, Embase and CNKI databases were searched for randomized controlled trials and case-control trials of helmet-assisted noninvasive ventilation in the treatment of ARF. The outcome measures included respiratory rate, intubation rate, complication rate, mortality rate and arterial blood gas analysis of the commonly used indicators (PaCO2/ PaO2 / pH). The results of the included studies' odds ratio (OR) and its 95% confidential interval (CI) were analyzed using Stata software. RESULTS: The results of the analysis showed that the in-hospital mortality, intubation rate and complication rate were all significantly decreased with the p-value less than 0.05, which was statistically significant. CONCLUSIONS: Helmet-assisted noninvasive ventilation can significantly reduce hospital mortality, intubation rate and complication rate, improving the survival rate and prognosis of patients with ARF.

Calculation of Likelihood Ratio for Identifying Half Sibling Relationship When Both Biological Mothers Participate.

ABSTRACT: Objective To derive the formulae for likelihood ratio （LR） calculation of half sibling relationships when both mothers participate. Methods Based on the fact that both biological mothers participate in the identification of half sibling relationship between the two individuals, test hypothesis for the identification of half sibling relationship was established. Conditional probability ratios of genetic evidence under null hypothesis and alternative hypothesis conditions were simplified, and then applied to a real case of half sibling relationship identification. At the same time, the LR of half sibling relationships under the assumption that only a single biological mother or none of the biological mothers participate were respectively calculated. Results In the cases of identification of half sibling relationship from same fathers, with no biological father involved, after the same genetic indicator test analysis, when both biological mothers participate in the identification, the accumulated LR value was higher than that of accumulated LR with only a single biological mother or no parents participating. Conclusion When the autosome STR test is used for the identification and analysis of half sibling relationship between two individuals, the calculation of LR is more simple, intuitive and operable with both mothers participating. The biological mothers should participate in the test as much as possible, otherwise the number of STR loci would need to be increased for a more specific conclusion.

[Risk for metastasis of lymph node between sternocleidomastoid and sternohyoid muscle in papillary thyroid cancer].

Objective: To analyze the risk factors for metastasis of lymph nodes between sternocleidomastoid and sternohyoid muscle (LNSS) in papillary thyroid cancer (PTC). Methods: Papillary thyroid cancer patients with clinically positive lateral lymph node metastasis (cN1) who underwent surgery including LNSS dissection between May 1, 2013 and May 31, 2016 at the Department of Head and Neck Surgery, Fudan University Shanghai Cancer Center were retrospectively studied. Logistic regression analysis was performed to evaluate possible clinicopathological factors related to LNSS metastasis. Results: In 85 patients, 54 patients (63.5%) showed LNSS in their surgical specimen, and 20 patients (23.5%) had pathologically positive LNSS metastasis. Patients with LNSS showed preoperatively higher levels of serum thyroid stimulating hormone (TSH) and thyroid peroxidase antibody (TPO-Ab) compared to patients only with fibrofatty tissues between sternocleidomastoid and sternohyoid muscle (P<0.05), and they also displayed a higher proportion of multifocality in ipsilateral thyroid lobe (P<0.05). Multi-factor analysis indicated that LNSS metastasis was correlated with original tumor size (OR=1.819, 95%CI 1.050-3.850, P=0.002) and Level Ⅳ lymph node metastasis (OR=2.190, 95%CI 1.132-2.334, P=0.005). Furthermore, the number of positive LNSS was tightly correlated to that of level Ⅳ lymph node metastasis(P<0.05). Conclusion: LNSS metastasis is occult but not quite rare in PTC. Patients with extensive lymph node metastasis in Level Ⅳhave a higher risk for metastasis of LNSS.

Evaluation of immunogenicity and protective efficacy of recombinant ptfA of avian Pasteurella multocida.

Avian Pasteurella multocida is the causative agent of fowl cholera, a disease much affecting the poultry industry. In order to study the efficacy of the recombinant subunit vaccine constructed with ptfA gene of avian P. multocida, the ptfA gene fragment amplified by PCR from avian P. multocida was cloned into the prokaryotic expression vector pET32a and the recombinant plasmid pET32a-ptfA was obtained. The pET32a-ptfA was expressed in Escherichiacoli BL21(DE3) and the target protein rPtfA was purified. The purified protein was then mixed with Freund's adjuvant and the recombinant subunit vaccine was obtained. Three groups of chickens labeled as rPtfA, attenuated live vaccine and PBS were vaccinated with the recombinant subunit vaccine, attenuated live vaccine and PBS, respectively. Serum antibodies, peripheral blood lymphocyte proliferation (PBLP) and interferon-γ (IFN-γ) level secreted by peripheral blood lymphocyte were tested. The immunized chickens were finally challenged with virulent avian P. multocida and the protection rate was counted. Indirect ELISA showed the levels of antibodies in rPtfA and attenuated vaccine groups were most significantly higher than the other groups (P<0.01), and the former was slightly lower than the latter. Peripheral blood lymphocyte proliferation experiments and IFN-γ experiments indicated that SI value and the levels of IFN-γ induced by ConA in the two vaccine groups were significantly higher than those of the PBS groups (P<0.01), and that the attenuated vaccine group was higher than the rPtfA group. The protection rates of rPtfA and attenuated live vaccines were 45% and 75%, respectively. The results indicated that the PtfA recombinant subunit vaccine was capable of improving the immunity level and inducing a protective effect for the vaccinated chickens, but it was barely satisfactory.

Cloning, characterization, and expression analysis of LGP2 cDNA from goose, Anser cygnoides.

Laboratory of genetics and physiology 2 ( LGP2: ) is a homologue of the retinoic acid inducible gene-I and melanoma differentiation associated gene 5 that lacks the caspase activation and recruitment domain required for signaling. It plays a pivotal role in host immune response. In this study, we cloned and characterized the full-length open reading frame ( ORF: ) sequence of LGP2 in the Qingyuan goose (Anser cygnoides) and evaluated the mRNA expression of this gene post infection with an H5N1 highly pathogenic avian influenza virus ( HPAIV: ). The full-length goose LGP2 ORF (2,028 bp) encoded a polypeptide of 675 amino acids. The deduced amino acid sequence contained 5 main overlapping structural domains-2 DEAD/DEAH box helicase domains, one conserved restriction domain of bacterial type III restriction enzyme, one helicase superfamily C-terminal domain and one C-terminal regulatory domain. Quantitative real-time PCR analysis indicated that goose LGP2 was constitutively expressed in all 19 investigated tissues, but the expression level was different among them. It was high expressed in the trachea, jejunum, bursa, kidney and heart, but low in the glandular stomach, lung, liver, spleen, crop and muscular stomach. A significant increase in the transcription of LGP2 was detected in the brain, spleen and lungs of geese post infection with H5N1 HPAIV versus uninfected tissues. These findings indicated that goose LGP2 was an important receptor that is involved in the host antiviral innate immune defense to H5N1 HPAIV in geese.

Detection of fetal sex chromosome aneuploidy by massively parallel sequencing of maternal plasma DNA: initial experience in a Chinese hospital.

OBJECTIVES: To evaluate the performance of a massively parallel sequencing (MPS)-based test in detecting fetal sex chromosome aneuploidy (SCA) and to present a comprehensive clinical counseling protocol for SCA-positive patients. METHODS: This was a retrospective study in a large patient cohort of 5950 singleton pregnancies which underwent MPS-based testing as a prenatal screening test for trisomies 21, 18 and 13, with X and Y chromosomes as secondary findings, in Southwest Hospital in China. MPS-based SCA-positive women were offered the choice of knowing whether their SCA results were positive and those who did commenced a two-stage post-test clinical counseling protocol. In Stage 1, general information about SCA was given, and women were given the option of invasive testing for confirmation of findings; in Stage 2, those who had chosen to undergo invasive testing were informed about the specific SCA affecting their fetus and their management options. RESULTS: Thirty-three cases were classified as SCA-positive by MPS-based testing. After Stage 1 of the two-stage post-test clinical counseling session, 33 (100%) of these pregnant women chose to know the screening test results, and 25 (75.76%) underwent an invasive diagnostic procedure and karyotype analysis, in one of whom karyotyping failed. In thirteen cases, karyotyping confirmed the MPS-based test results (two X0 cases, seven XXX cases, three XXY cases and one XYY case), giving a positive predictive value of 54.17% (13/24 cases confirmed by karyotyping). After post-test clinical counseling session Stage 2, seven women chose to terminate the pregnancy: one X0 case, two XXX cases, the three XXY cases and the single XYY case. Six women decided to continue with pregnancy: one X0 case and five XXX cases. CONCLUSION: Our study showed the feasibility of clinical application of the MPS-based test in the non-invasive detection of fetal SCA. Together with a two-stage post-test clinical counseling protocol, it leads to a well-informed decision-making procedure.

Chronological changes of delayed-type hypersensitivity in mice immunised with testicular germ cells alone.

Experimental autoimmune orchitis (EAO), comprising a breakdown of the testicular immune privilege, is one of the models of immunological male infertility. EAO is characterised by CD4 + T-cell-dependent lymphocytic inflammation and augmented delayed-type hypersensitivity (DTH) against testicular antigens. We previously established an EAO model in mice by immunisation with viable syngeneic testicular germ cells (TGC) alone. However, the sequential change of DTH during development of this EAO has not been analysed yet. In this study, the DTH response during TGC-induced EAO was investigated by the injection of syngeneic TGC protein into the ears of mice. The results showed that a significant DTH response was observed on injection of 20 µg TGC protein, but not on that of 0.2 or 2 µg TGC protein. Also, the level of the DTH response to 20 µg TGC protein was highly relevant to the pathology of EAO development. These results indicate that the DTH response on injection of 20 µg TGC protein into the ears of mice is effective for predicting the pathology of EAO development.

Structure of thymic cysts in congenital lymph nodes-lacking mice.

The ALY (aly/aly) mouse, a mutant of the C57BL/6j strain, has a severe immunodeficiency because of immature development of the immune organs. Both lymph nodes and Peyer's patches are lacking and both the thymus and spleen are small. Previous microscopical observation of their thymus glands revealed the presence of an indistinct border between the cortex and medulla, the absence of Hassal's corpuscles and the reduction of the medullary epithelial cell population. However, other microscopical findings for these glands have not yet been reported. In the present study, we performed light and electron microscopical observation of the thymus and found the consistent presence of extremely irregular shaped cystic cavities lined by microvilli-bearing epithelium in the medulla. The cysts comprised ceca and did not open into adjacent capillaries, although they contained some lymphocytes and macrophages in their lumens. In the thymus glands of normal C57BL/6j mice, only some small cysts oval in shape could be inconspicuously found in the medulla. Therefore, the thymic cysts may normally regress during thymic development, however, in ALY mice, the cysts may remain because of the organ immaturity.

Simultaneous determination of two active components in compound aspirin tablets using principal component artificial neural networks (PC-ANNs) on NIR spectroscopy.

A method for simultaneous, non-destructive analysis of aspirin and phenacetin in compound aspirin tablets with different concentrations has been developed by principal component artificial neural networks (PC-ANNs) on near-infrared (NIR) spectroscopy. In PC-ANNs models, the spectra data were first analyzed by principal component analysis (PCA). Then the scores of the principal compounds (PCs) were chosen as input nodes for input layer instead of the spectra data. The artificial neural networks (ANNs) models using the spectra data as input nodes were also established, which were compared with the PC-ANNs models. Four different preprocessing methods (first-derivation, second-derivation, standard normal variate (SNV) and multiplicative scatter correction (MSC)) were applied to NIR conventional spectra. The result shows the first-derivative model of PC-ANNs multivariate calibration has the lowest training errors and predicting errors. The concept of the degree of approximation was introduced and performed as the selective criterion of the optimum network parameters.

[Effect of methionine supplementation on the selenium bioavailability in rats fed on grains from Keshan disease endemic area].

In order to study the effect of dietary methionine on tissue selenium and glutathione peroxidase activity(GPX) in rats fed on grains from Keshan Disease endemic area, a nonpurified selenium (Se) deficient diet (containing Se 0.007 mg/kg diet) taken from Keshan Disease endemic area was supplemented with selenomethionine (SeMet) to provide selenium content in diet for 0.007, 0.06 and 0.50 mg/kg and added or not added with DL-methionine (Met) 4 g/kg to make 6 kinds of diet. Fifty four male weanling Wistar rats were randomly divided into 6 groups, consuming each one of the 6 diets for 8 weeks. The Se content and GPX activity in tissues of animals with Met supplementation were compared with those without Met supplementation. The results showed that adding Met did not result in significant changes of tissue Se content and GPX activity in animals having dietary Se of 0.007 mg/kg, except for lower muscle Se content. However, in animals having dietary Se of 0.06 mg/kg and supplementing Met resulted in selenium redistribution in tissues-decrease of Se content in muscle, increase of Se content in liver and blood and significant elevation of GPX activity in all tissues. In animals having methionine-added and dietary Se of 0.50 mg/kg, tissue Se content declined to various extent, while GPX activity remained unchanged. The results suggest that SeMet (main chemical form of Se in cereals) is preferentially incorporated into body protein when dietary methionine is limited. Once Met is supplemented, dietary SeMet would provide more Se for the syntheses of GPX and other selenoproteins. The results further suggest that marginal deficiency in sulfur-containing amino acids in the diet from Keshan Disease endemic area might be an additional factor for the development of Keshan Disease under selenium deficiency.

[Studies on the determination of serum thymic factor by capillary zone electrophoresis].

The determination of serum thymic factor (FTS) by capillary zone electrophoresis was described. The optimum ionic concentration and the pH value of buffer system was examined through orthogonal analysis. The optimum conditions found to be 0.05 mol/L sodium tetraborate, 0.01 mol/L phosphate buffer at pH 8.70, working voltage at 12 kV and wavelength at 200 nm. The recoveries for FTS standards added to the bovine serum albumin and serum samples were measured. The values of the relative standard deviation(RSD), recovery and detection limit were 7.62%, 80.28% and 40 ng/ml respectively. The development of this method provide another way for studying the relations of zinc and thymic factor and investigating the effect of zinc on immune function.

An ethanol inducible gene switch for plants used to manipulate carbon metabolism.

Many transgenic plant studies use constitutive promoters to express transgenes. For certain genes, deleterious effects arise from constant expression in all tissues throughout development. We describe a chemically inducible plant gene expression system, with negligible background activity, that obviates this problem. We demonstrate its potential by showing inducible manipulation of carbon metabolism in transgenic plants. Upon rapid induction of yeast cytosolic invertase, a marked phenotype appears in developing leaves that is absent from leaves that developed before induction or after it has ceased.

Mutations in the Fas antigen in patients with multiple myeloma.

Programmed cell death, or apoptosis, is well documented as a physiological means of eliminating activated lymphocytes and maintaining immune homeostasis. Apoptosis has also been implicated in the targeting of tumor cells by cytotoxic T lymphocytes and natural killer cells. One of the two primary mechanisms used in cell-mediated cytotoxicity is the Fas/FasLigand system. Activated or transformed cells expressing the Fas antigen on their surface are susceptible to killing by immune effector cells that express the Fas ligand. Many neoplastic cells, including those derived from patients with multiple myeloma, express Fas antigen on their surface, but do not undergo apoptosis in response to antigen crosslinking. One possibility for the lack of Fas-mediated apoptosis includes mutations in the Fas antigen. Loss of function mutations in the Fas antigen have been associated with congenital autoimmune disease in humans, and have been defined as the genetic defect the in lpr mice. Mutations in the Fas antigen have not been previously described in cancer patients. In this study, we show that mutations occur in the Fas antigen which may cause loss of function and contribute to the pathogenesis of the neoplastic disease, multiple myeloma. Using reverse transcriptase-polymerase chain reaction (RT-PCR), single-stranded conformation polymorphism (SSCP) analysis, and DNA sequencing, we examined the cDNA structure of the Fas antigen in 54 bone marrow (BM) specimens obtained from myeloma patients. Six patient specimens (11%) did not express detectable levels of Fas antigen mRNA. Of the 48 BM specimens which did express Fas antigen, 5 (10%) displayed point mutations. All of the mutations identified were located in the cytoplasmic region of the Fas antigen known to be involved in transduction of an apoptotic signal. Two separate individuals demonstrated an identical mutation at a site previously shown to be mutated in the congenital autoimmune syndrome, ALPS. One patient exhibited a point mutation at a site only two amino acids removed from the documented lesion of the lprcg mouse. Although the functional status of these point mutations remains to be determined, we propose that Fas antigen mutations may contribute to the pathogenesis and progression of myeloma in some patients.

[The Chinese total diet study in 1992--macro elements and trace elements].

Ten macro elements and trace elements were directly determined in the 1992 total diet study conducted in four districts of China from north to south. The intake of these elements was evaluated. The levels of intake for seasonal variation and for different age groups were reported. The results showed that the intakes of Ca, P, K, Mg, Zn, Se and Cu were low in children. The intake of Ca, P and Zn was insufficient for adults. The intakes of other seven elements reached or approached their RDA levels, but there were big differences among the 4 districts. The relationships between intakes of Ca/P and Na/K as well as Fe and Cu and iron difficiency anemia were discussed.