Amino Acid Sequence ; Genetic Structures ; Genetic Variation ; Genome, Viral ; Molecular Sequence Data ; SARS Virus ; genetics

Severe acute respiratory syndrome(SARS), caused by SARS- associated coronavirus(SCV), is the first severe infectious disease in this century. SARS is pathologically characterized by interstitial exudative inflammation of lung with the formation of hyaline membrane in acute phase. Haemorrhagic inflammation exists in extrapulmonary organs. Clinical diagnosis is a dynamic process and includes the suspected case, probable case and definite case. Diagnostic standard of SARS will be revised with further understanding of the disease. Chinese term of SARS has been recommended in the paper.
Humans ; Severe Acute Respiratory Syndrome ; diagnosis ; epidemiology ; pathology ; Terminology as Topic

AIMTo observe the effects of Ca2+ -activated K+ channel of primary cultured fetal SD rat cortex neurons in the veratridine triggered neuronal damage.

METHODSThe patch clamp technique of cell-attach and inside-out mode for these two kinds of single channel recordings were used.

RESULTSExtracellular veratridine activated the Kca. In Ca2+ bath solution of cell-attach mode, Vp + 30 mV, when the concentration (micromol/L) of veratridine were 15,25,50 and 75, the open probabilities of the channel were 0.014 +/- 0.003, 0.085 +/- 0.010, 0.132 +/- 0.016 and 0.059 +/- 0.006 (P < 0.01) respectively. It appeared concentration-dependent within 50 micromol/L veratridine. In Ca2+ free bath solution of cell-attach mode, Vp = +50 mV, when the concentration (micromol/L) of veratridine were 15, 40,60 and 100, the open probabilities of the channel were 0.014 +/- 0.010, 0.113 +/- 0.006, 0.141 +/- 0.004 and 0.295 +/- 0.009 (P < 0.05) respectively. In the 6 cases of inside-out mode patch clamp, Vp = +40 mV, when the concentration of veratridine were 0, 25 micromol/L and 50 micromol/L, the open probabilities of the channel were 0.011 +/- 0.008, 0.010 +/- 0.010 and 0.012 +/- 0.007 (P > 0.05) respectively. There were no significant difference on open probabilities, average open/close times and amplitudes at different intracellular veratridine concentration.

CONCLUSIONVeratridine can affect the activation of the Kca channel through regulating the concentration of cytoplasmic free Ca2+. The opening of Kca activated by increase of intracellular Ca2+ during the early stage of anoxia may be a protection reaction of ischemic neurons.

Animals ; Animals, Newborn ; Calcium ; metabolism ; Cells, Cultured ; Neurons ; cytology ; drug effects ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Calcium-Activated ; metabolism ; Rats ; Rats, Sprague-Dawley ; Veratridine ; pharmacology

OBJECTIVETo systemically analyze the differentially expressed genes from normal mucosa and carcinoma of colon obtained by SSH method.

METHODSAn automatic platform for the analysis of nucleotides based on Linux was constructed, and one of the three subtracted libraries from SSH (T -N) was systematically analyzed by this platform. Part of the results was verified by semi- quantity RT-PCR.

RESULTThe automatic platform for the analysis of nucleotides based on Linux was successfully constructed. There were 15 contigs from the subtracted T-N libraries, among which 2 had no match with known genes in the GenBank. The expressions of genes Homo Sapiens thymosin beta 4 (THY) and Homo Sapiens HbxAg transactivated protein 2 (XTP) had trends of increase with the progress of normal tissue-adenoma-adenocarcinoma when verified by semi- quantity RT-PCR.

CONCLUSIONLinux-based automatic platform provides an efficient way to systematically analyze the nucleotide sequences, which may be used in study on the mechanism of colorectal carcinogenesis.

Colon ; metabolism ; Colonic Neoplasms ; genetics ; Computational Biology ; Female ; Gene Expression Profiling ; Humans ; Intestinal Mucosa ; metabolism ; Male ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo examine whether the enhanced expression of CD40L cDNA on murine ovarian cancer (OVHM) cells could induce the secretion of Th1 cytokines from dendritic cells (DC).

METHODSOVHM cells were transfected with the full-length mouse CD40L cDNA by lipofectamine 2000 and then G418 resistant cells as positive cells were selected. They were examined for their expression of CD40L with flow cytometry. Bone marrow cells were firstly depleted of erythrocytes, macrophages, T and B cells with PE-conjugated magnetic beads, and then cultured in 10% FCS RPMI 1640 medium supplemented with recombinant mouse GM-CSF and IL-4 for 10 days. PKH67-labeled tumor cells were cultured with DC, and then the stained cells were analyzed for the expression of MHC-I, MHC-II, CD80, CD86, CCR7 in DC with flow cytometry. The expression of p40, p19, p35, p28, EBI3 subunits, IL-18, IFN-gamma, Mig gene in cocultured DC-tumor cells were detected by RT-PCR.

RESULTSThe CD40L cDNA was successfully transfected into OVHM cells. Bone marrow-derived DCs, when cultured with CD40L/OVHM, formed clusters with the tumors and showed an upregulated expression of MHC- I, MHC-II, CD80, CD86, CCR7. Expression of the IL-12, IL-23, IL-27, IL-18, interferon-gamma (IFN-gamma) and Mig (monokine induced by IFN-gamma) genes was induced in the DCs that were cultured with CD40L/OVHM but not with OVHM cells.

CONCLUSIONThese data directly showed that the expression of CD40L on ovarian cancer cells facilitates the interaction between DCs and tumors, enhances the maturation of DCs, induces secretion of Th1 cytokines, especially for IL-12, IL-23 and IL-27, which maybe one of the possible antitumor mechanism for CD40L-transfected ovarian cancer cell line.

Animals ; CD40 Ligand ; genetics ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Coculture Techniques ; Cytokines ; secretion ; DNA, Complementary ; genetics ; Dendritic Cells ; cytology ; metabolism ; Female ; Interleukin-12 ; secretion ; Interleukin-23 ; secretion ; Interleukins ; secretion ; Mice ; Ovarian Neoplasms ; metabolism ; pathology ; Th1 Cells ; secretion ; Transfection

OBJECTIVETo examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murine ovarian cancer OVHM cells (CD40L-OVHM) injected into the spleen on liver metastasis in mice.

METHODSOVHM cells were inoculated into the spleen of 6 to 8 week-old female B6C3F1 (C57BL/6N x C3H/He) mice. The established liver metastasis was identified by histopathology (HE staining). OVHM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CD11c in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-gamma, TNF-alpha, IL-12, IL-4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded.

RESULTSThe mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11c and the specific killing rate in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-gamma, TNF-alpha and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo-OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly (p < 0.05). The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group (P < 0.05).

CONCLUSIONThe data directly demonstrate that the expression of CD40L in ovarian cancer cells (CD40L-OVHM) can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune function of peripheral blood cells and the immune balance between Th1 cells and Th2 cells, which maybe the possible mechanism induced by CD40L in mice inhibiting the development of liver metastasis.

Animals ; CD11c Antigen ; metabolism ; CD40 Ligand ; genetics ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; DNA, Complementary ; genetics ; Dendritic Cells ; cytology ; Female ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-4 ; metabolism ; Liver Neoplasms ; pathology ; secondary ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Ovarian Neoplasms ; metabolism ; pathology ; Spleen ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVENo optimal prophylactic protocol of hepatitis B immunoglobulin (HBIG) combined with nucleos(t)ide analogue for HBV recurrence has been established yet. By investigating the alterations of HBV markers in HBV related liver disease patients, recipients of a liver transplant, under lamivudine or/and HBIG prophylaxis, we aim to explore the possible HBV recurrence mechanism involved and to find a new option in the prophylaxis of HBV recurrence and to tailor individualized therapy.

METHODSSerial liver biopsy specimens and sera were obtained intraoperationally and at definite time points during follow-up. ELISA and chemiluminescent microparticle immunoassay, HBV DNA fluorescent quantification, immunohistochemistry staining and HBV DNA in situ hybridization were performed. Alterations of HBV markers in specimens of 96 liver transplant recipients were investigated retrospectively.

RESULTSAll 17 cases had HBV recurrence (median 37 months) which occurred in the follow-up period after liver transplantation. The overall actual HBV recurrence rate at 2 years was 22% with a significant difference between that of the active and inactive groups (P<0.05); 82.4% HBV recurrence took place within the first 3 years after the operation, and the recurrence ratio of first 3 years to 3 years later after transplantation was 4.7 (P<0.01). The HBV DNA positive patients accounted for 78.6% of the total number of recurrences within the first 3 years. HBcAb and HBeAb positive rates went down with time, but their positivity remained.

CONCLUSIONHBV recurrence happens after liver transplantation. In inactive HBV replicative patients with strictly combined prophylaxis and availability of other medications and using 3 years after liver transplantation as a point of time, we think that tapering down the dosage of HBIG and tailoring individualized treatment methods based on virological and immunological situations of each recipient are worth trying.

Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Child ; Female ; Hepatitis B ; drug therapy ; Humans ; Immunoglobulins ; therapeutic use ; Lamivudine ; therapeutic use ; Liver Transplantation ; Male ; Middle Aged ; Postoperative Period ; Prognosis ; Retrospective Studies ; Secondary Prevention ; Treatment Outcome ; Young Adult

BACKGROUNDThe offspring of women with gestational diabetes mellitus (GDM) are prone to macrosomia. However, birth weight is difficult to be correctly estimated by ultrasound because of fetal asymmetric growth characteristics. This study aimed to investigate the correlations between fetal hemodynamics, fetal growth indices in late pregnancy, and birth weight in GDM.

METHODSA total of 147 women with GDM and 124 normal controls (NC) were enrolled in this study. Fetal hemodynamic indices, including the systolic/diastolic ratio (S/D), resistance index (RI), pulsatility index (PI) of umbilical artery (UA), middle cerebral artery (MCA), and renal artery (RA), were collected. Fetal growth indices, including biparietal diameter (BPD), head circumference (HC), abdominal circumference (AC), and femur length, were also measured by ultrasound. Birth weight, newborn gender, and maternal clinical data were collected.

RESULTSThe independent samples t-test showed that BPD, HC, and AC were larger in GDM than in NC (P < 0.05). Fetal hemodynamic indices of the UA and MCA were lower (P < 0.05), but those of the RA were higher (P < 0.001) in GDM than in NC. Birth weight was higher in GDM than in NC (P < 0.001). Pearson's correlation analysis showed that hemodynamic indices of the UA were negatively correlated with birth weight, BPD, HC, and AC in both groups (P < 0.05). MCA (S/D, PI, and RI) was negatively correlated with birth weight, HC, and AC in GDM (r = -0.164, -0.206, -0.200, -0.226, -0.189, -0.179, -0.196, -0.177, and - 0.172, respectively, P< 0.05), but there were no correlations in NC (P > 0.05). RA (S/D, PI, and RI) was positively correlated with birth weight in GDM (r = 0.168, 0.207, and 0.184, respectively, P< 0.05), but there were no correlations in NC (P > 0.05).

CONCLUSIONFetal hemodynamic indices in late pregnancy might be helpful for estimating newborn birth weight in women with GDM.

Adult ; Birth Weight ; physiology ; Cerebral Arteries ; physiology ; Diabetes, Gestational ; physiopathology ; Female ; Fetal Development ; physiology ; Hemodynamics ; physiology ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Pregnancy Outcome ; Renal Artery ; physiology ; Ultrasonography, Prenatal ; Umbilical Arteries ; physiology

Objective】 To explore the clinical application of the fifth edition of BI- RADS for the category of suspicious calcification. 【Methods】 Mammograms of patients with suspicious calcification from May 2012 to May 2017 were reviewed retrospectively. The morphology（amorphous，coarse heterogeneous，fine pleomorphic，fine linear or fine linear branching）and distribution（diffuse，regional，grouped，linear，segmental）of suspected calcification according to the fifth edition of the BI- RADS were analyzed. Taking pathology as the gold standard，the positive predictive value（PPV）of morphology，distribution and combination of morphology and distribution was calculated and compared with the PPV range of the fifth edition of BI- RADS. 【Results】 A total of 170 patients were included in the study，116 were benign，54 were malignant，and PPV was 31.8%（54/170）. In terms of morphology，amorphous，coarse heterogeneous， fine pleomorphic calcification were category 4B（10%

Callicarpa nudiflora,which is a big brand of Li nationality medicine with Hainan characteristics,has the effects of dissolving stasis,hemostasis,anti-inflammatory and antibacterial. At present,there is a lack of information about the reference genome of C. nudiflora. The study of the genome size,heterozygosity rate and characteristics of SSR of C. nudiflora,can provide an effective basis for the formulation of the whole genome de novo sequencing strategy and development of SSR molecular markers of C. nudiflora. To realize this purpose,high throughput sequencing platform Illumina Hiseq was used to sequence the genome structure of C. nudiflora and K-mer analysis was applied to estimate genome size,repeat sequences and heterozygosity rate. Simple-sequence repeat( SSR) loci that are suitable as markers were identified by MISA software. The results showed the estimated genome size of C. nudiflora was 822. 43 Mb,with a 0. 85% heterozygosity rate and 71. 67% repeats,and the GC content of genome was about 49. 20%. Therefore,C. nudiflora belongs to a complex genome with high heterozygosity and repetition. SSR molecular genetic markers were analyzed in the genome sequence,and a total of 206 049 SSRs were identified,among which mono-nucleotide,di-nucleotide and tri-nucleotide repetitive motifs summed up to 198 993,accounting for 96. 57% of the total SSRs. Among the 2-6 nucleotide repeats,AT/AT,AAT/ATT,AGCC/CTGG,AAAAT/ATTTT and AGATAT/ATATCT have the largest number,respectively. This report represents the first genome-wide characterization of C. nudiflora,and provides a reference for the construction of the library for the fine sequencing of the genome,and a molecular basis for the development of SSR molecular markers as well as for the protection and utilization of gene resources.
Callicarpa/genetics* ; Genetic Markers ; Genome, Plant ; Microsatellite Repeats ; Polymorphism, Genetic