Objective　Constructed a nanopore sequencing traceability technology system based on the globally unified DENV-1 genotyping collaborative monitoring strategy and applied it preliminarily to the serum sample testing of patients with dengue fever. Methods Utilizing the DENV-1 whole genome sequences set in the Global Integrated Sequence and Genotyping Database for DENV (GISDD v1.2.1), the specific targeted multiplex PCR primer pairs were screened, designed, and evaluated using MEGA 7.0, Prime-Blast, MPprimer, and GISDDrPrimer softwares. The RNA from the serum samples was extracted and subjected to reverse transcription. The 16 pairs of primers were designed into 2 primer pools for multiplex PCR amplification of the target genome sequences. The library was constructed using magnetic bead purification and the SQK-LSK109 kit according to the operational procedure, followed by MinION sequencing. The sequencing data were analyzed and DENV-1 genomes were assembled for traceability analysis using the GISDD genetic typing platform. Results Sixteen pairs of DENV-1 primers with high coverage to the DENV-1 genomes were designed and identified, establishing a DENV-1 specific multiplex PCR-based nanopore high-throughput sequencing system. It was preliminarily applied to the whole-genome detection of serum samples from 13 patients with dengue fever in Guangzhou. The median of average reads length was 1 668.00 bp, the median of average reads quality was 9.30, the median alignment rate for the reference genome was 97.90%, and the median length aligned to the reference genome NV46 was 1 677.90 bp, with a median genome depth of 12 886.40×. The subsequent genotyping and tracing analyses using GISDD revealed that 9 genomes belonged to DENV-1 clade 1E1, 3 to 1L1, and 1 to 5C1. Conclusions The nanopore sequencing-based DENV-1 whole-genome detection system contextualizing within GISDD described here is characterized by its stability and reliability, enabling the acquisition of high-quality DENV-1 genomes. This approach offers an efficient technical solution for the rapid identification and traceability of DENV genomes during dengue outbreaks, thereby establishing a solid foundation for developing a global collaborative surveillance and control system for dengue.

OBJECTIVETo study the effects of Chaiqiyigan Granula (CG) and Taxol on the growth of hepatocellular carcinoma (HCC) xenografts and expression of Bax, p53 and VEGF in nude mice.

METHODSWhole-body fluorescence imaging was used to visualize the growth of HCC in nude mice bearing hepG2/EGFP cell xenograft. Immunohistochemistry was used to determine the content of Bax, p53 and vascular endothelial growth factor (VEGF) in the tumor tissues.

RESULTSCompared with normal saline, Taxol alone and in combination with CG significantly inhibited the growth of HCC xenografts in the nude mice. The combined treatment with CG and Taxol produced a stronger inhibitory effect on the tumor growth than Taxol alone in the third and fourth weeks. The volume and weight of the xenografts were decreased in the combined treatment group compared with those in saline treatment group. CG combined with Taxol increased the expression of Bax and reduced the expression of p53 and VEGF in the tumor xenografts.

CONCLUSIONCG can enhance the inhibitory effects of Taxol on the growth of HCC xenografts, and this effect is related to the up-regulation of Bax and down-regulation of p53 and VEGF expression in the tumor.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Mice ; Mice, Nude ; Optical Imaging ; Paclitaxel ; pharmacology ; Tumor Suppressor Protein p53 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays ; bcl-2-Associated X Protein ; metabolism

Objective To renovate angiography in identifying portal vein anatomy during transjugular intrahepatic portosystemic shunt (TIPS) procedures,saving the time of TIPS procedures,decreasing the risk of the complications of the post-procedure.MethodsThe difference between the Wedge hepatic venography with Carbon Dioxide in 6 cases and Inferior Mesenteric artery angiography in 7 cases during TIPS procedures were compared in the identification of portal vein anatomy.The quality of images,their effects on the procedures,the complications and the recovery post-procedure were evaluated.Results Using CO2,the portal veins were opacified in all 6 cases.TIPS procedures succeeded in all cases except 1 case because of poor coagulation function.Using Inferior Mesenteric artery angiography,the portal veins were opacified in al1 7 cases.TIPS procedure succeeded in all cases except 1 case because of chronic portal occlusion.Puncture-site hematoma occurred in 1 case after TIPS procedure.ConclusionWedge hepatic venography with Carbon Dioxide is superior,safer and more convenient than Inferior Mesenteric Artery angiography in identifying portal vein anatomy during TIPS.

Objective To identify one runny mucoid-like Gram-negative bacteria with pink pigment isolated from clinical pus sample. Methods The pus sample was aseptically extracted from a deep lesions of one patient, then stored in Amies medium at room temperature for transportation. One sheep blood plate and one chocolate plate were used to detect the possible pathogens from the specimens. After inoculation, the plates were placed in a humidified incubator with 5% CO2 at 35 ℃. To identify the obtained isolates, we used the commercial Vitek2 and API systems, combining some traditional morphological examination and classical biochemical and physiological characteristics. For pure cultures, the cellular fatty acids were extracted, methylated, and determined by gas chromatography method. The 16S rRNA gene was amplified and sequenced by a commercial broad-spectrum PCR primers. The phylogenetic tree based on 16S rRNA gene was constructed by Mega 4.1 software using the neighbour-joining methods with 1 000 bootstrap replications. Results One runny mucoid-like Gram-negative bacterium, named K8756, was isolated both on sheep blood and chocolate plates after 72 h incubation. The API 20NE profile was 1245045 after a 3-day culture, which would be identified as Ochrobactrum anthropi with a good confidence of 98% probability. It was identified as Ralstonia pickettii and Bordetella bronchiseptica by VITEK 2 GN kits. However, further comparative 16S rRNA gene sequences showed that strain K8756 was closely related to the valid published Roseomonas mucosa MDA 5527 with 100% identity. Colonial morphologic features, phenotypic characteristics and major cellular fatty acid composition were also with high similarity to Roseomonas mucosa. Conclusions Strain K8756( = GIMCC 1.0030 ) is identified as Roseomonas mucosa by the polyphasic phenotypic and genotypic characteristics. The comparative analysis based on 16S rRNA gene sequences is a useful method for identifying the problematic and newly named bacteria.

Objective:To research the influence and mechanism of Toufeng capsule (TFC) on the concentratioin of blood active substance and single amine neurotransmitter in the brain and blood of magraine mice.Methods: We chose animal disease model of migraine made by hypodermic nitroglycerin and measured the concentration of 5 HT. NE and DA in the rat's brain and blood by means of enzymeimmunoassay experimental methods.Results: TFC could obviously increase the concentration of 5 HT and NE in the bloold and of 5 HT, DA and NE in the brain.Conclusions: TFC might improve neurosystematic regulation functions and enhance the neurotransmitters' secretion and metabolism.