Computerised modified paramedian approach technique versus conventional midline approach technique of lumbar puncture: a randomised control trial protocol.

INTRODUCTION: The lumbar puncture (LP) technique is widely used for diagnostic and therapeutic purposes. In recent years, the paramedian approach technique (PAT) has gained increasing interest due to its advantages over the conventional midline approach technique (MAT) that has been traditionally employed in clinical practice for LP. However, there have been inconsistent discussions regarding the efficacy of different LP techniques. Based on digital virtual human and computer simulation techniques, a new approach called computerised modified PAT (CMPAT) was proposed. Therefore, the aim of this study is to discuss a randomised controlled trial (RCT) protocol to investigate and compare the effects of CMPAT and MAT in patients undergoing LP. METHODS AND ANALYSIS: We will conduct a prospective, multicentre RCT. The study will recruit 84 patients aged 18-99 years who require LP. Participants will be randomly assigned to either the CMPAT treatment group (group A) or the MAT treatment group (group B). The primary outcome measure will be the number of needle insertion attempts required for a successful LP. Secondary outcomes will include the puncture success rate, pain assessment in the back, head, and legs, and the occurrence of complications. The measurement of these secondary outcomes will be taken during the procedure, as well as at specific time points: 30 min, 6 hours, 1 day, 3 days, 7 days, 2 weeks and 4 weeks after the procedure. Pain levels will be assessed using a Numerical Rating Scale. ETHICS AND DISSEMINATION: Ethical approval (2022YF052-01) has been obtained from the Ethics Committee of Fujian Medical University Union Hospital, Fuzhou, China. The research findings will be published in an international peer-reviewed scientific journal and presented at scientific conferences. TRIAL REGISTRATION NUMBER: ChiCTR2300067937.

Expression of rice gall dwarf virus outer coat protein gene (S8) in insect cells.

To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity, its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system. The S8 gene was subcloned into the pFastBac™1 vector, to produce the recombinant baculovirus transfer vector pFB-S8. After transformation, pFB-S8 was introduced into the competent cells (E. coli DH10Bac) containing a shuttle vector, Bacmid, generating the recombinant bacmid rbpFB-S8. After being infected by recombinant baculovirus rvpFB-S8 at different multiplicities of infection, Sf9 cells were collected at different times and analyzed by SDS-PAGE, Western blotting and immunofluorescence microscopy. The expression level of the P8 protein was highest between 48-72 h after transfection of Sf9 cells. Immunofluorescence microscopy showed that P8 protein of RGDV formed punctate structures in the cytoplasm of Sf9 cells.

Constituents from the seeds of Brucea javanica with inhibitory activity of Tobacco mosaic virus.

Three new constituents were obtained along with 10 known compounds from the seeds of Brucea javanica. The structures of these compounds were determined based on spectral and chemical evidence. These new compounds included a monoterpenoid glycoside and two sesquiterpenes. Bioactivity screening of these constituents showed that compounds 1, 3, 8, 9, and 13 with obvious activities in inhibiting multiplication of the Tobacco mosaic virus.

Antiphytoviral activity of bruceine-D from Brucea javanica seeds.

BACKGROUND: Brucea javanica (L.) Merr. is widely distributed throughout the southern parts of China and has been used in traditional medicine to treat a variety of diseases. The objective of the present study was to identify the active antiphytoviral compound in the seeds of B. javanica and evaluate the inhibitory activity of the compound against plant virus. RESULTS: Bioassay-guided fractionation of the most active extract from the seeds led to the isolation of an antiphytoviral compound which was identified as bruceine-D by conventional spectroscopy methods. The compound exhibited significant inhibitory activity against the infection and replication of tobacco mosaic virus (TMV), with IC(50) values of 13.98 and 7.13 mg L(-1) respectively. The compound also showed a strong inhibitory effect on the infectivity of potato virus Y (PVY) and cucumber mosaic virus (CMV). Furthermore, the compound could effectively inhibit systemic TMV infection in the host tobacco plant under glasshouse conditions. CONCLUSION: The results suggested that bruceine-D from Brucea javanica may have the potential to be used as a natural viricide, or a lead compound for new viricides.

[Effects of microcystins on expression of apoptosis-associated hepatocyte genes].

OBJECTIVE: To deeply explore the effects of microcystins (MC-LR) on Bax and Bcl-2 during the course of MC-LR promoting liver tumor. METHODS: applied to set up the animal model, and the effect of MC-LR promoting liver tumor was evaluated by the Albertgamma-GT methods. And then, the immunohistochemical technique, RT-PCR and image analysis were used to study the expression of the Bcl-2 and Bax during the course of promoting tumor. RESULTS: (1) MC-LR might enhance the positive reaction rate of GGT. The positive reaction rate of GGT in DEN + pure toxin group was 100%, it was significantly higher than the DEN control group 22.22% (P < 0.05). (2) The intension and areas of the protein expression of Bcl-2 in DEN + pure toxin group were 0.0977 and 0.0315, and in DEN control group were 0.0460 and 0.0205, respectively. The expression level of Bcl-2 protein in DEN + pure toxin group were significantly higher than in DEN control group (P < 0.05). Simultaneously, the protein expression of Bax was significantly decreased by MC-LR (P < 0.05). The intension and areas of the expression of Bax in DEN + pure toxin group were 0.0283 and 0.0073, and in DEN control group were 0.0655 and 0.0244 respectively. (3) The mRNA expression of Bcl-2 was significantly increased by MC-LR. The intension of Bcl-2 mRNA expression in DEN + pure toxin group was 2.244, being significantly higher than in the other groups (P < 0.05). However, the mRNA expression of Bax showed no significant difference between DEN + pure toxin and the other groups. CONCLUSION: The expression change of Bcl-2 and Bax should possibly play an important role in the course of MC-LR promoting liver tumor.

Structure-activity relationship of triterpenes and triterpenoid glycosides against tobacco mosaic virus.

A new activity of triterpenes and triterpenoid glycosides against phytovirus has been found. Based on the anti-TMV replication activity in vivo, 47 triterpenes and triterpenoid glycosides were screened through the leaf-disk method together with indirect ELISA. Structure-activity relationships were analyzed according to the results of bio-activity screening. Besides, the EC(50) values of those compounds having higher activities were measured.

[Current situation and prospect of studies on genus Umbravirus].

Umbraviruses are a group of imperfectly characterized plant viruses, which are distinguished from most other viruses by their genomes lack of a gene for coat protein (CP) , and as a result umbraviruses do not form conventional virus particles. Umbraviruses are mechanically transmissible, and can be aphid transmitted in the persistent manner by an unrelated assistor virus, which is always a member of the family Luteoviridae . In nature, each umbravirus depends for survival on one particular luteovirus. The genus Umbravirus comprises seven distinct virus specieses and three tentative members. Only Tobacco bushy top virus (TBTV) has been reported in China as an umbravirus. Tobacco bushy top disease, caused by TBTV and its helper, Tobacco vein distorting virus(TVDV), which resulted in severe tobacco losses in western of Yunan. Umbraviruses had a restricted host range in nature, and their infectivity and longevity in vitro are not so stable. Plants infected umbraviruses contain abundant double-stranded RNA (dsRNA), and some umbraviruses possess one or more additional dsRNA species associated with the presence of a satellite RNA. The genomes of the umbraviruses consist of one linear segment of positive sense single-stranded RNA(ssRNA), and the nucleotide sequences possess ORFs for four potential non-structural protein products. The umbravirus-encoded ORF3 proteins play essential roles in stabilization of viral RNA and mediation of its long-distance movement. The current research progresses have been reviewed detailly, and the future research tendency and research fields about umbraviruses and umbravirus-caused diseases are put forward.

Molecular characterization of a new lectin from the marine alga Ulva pertusa.

A new lectin, named UPL1, was purified from a green alga Ulva pertusa by an affinity chromatography on the bovine-thyroglobulin-Sepharose 4B column. The molecular mass of the algal lectin was about 23 kD by SDS-PAGE, and it specifically agglutinated rabbit erythrocytes. The hemagglutinating activity for rabbit erythrocytes could be inhibited by bovine thyroglobulin and N-acetyl-D-glucosamine. The lectin UPL1 required divalent cations for maintenance of its biological activity, and was heat-stable, and had higher activity within pH 6-8. The N-terminal amino acid sequence of the purified lectin was determined (P83209) and a set of degenerate primers were designed. The full-length cDNA of the lectin was cloned by rapid amplification of cDNA ends (RACE) method (AY433960). Sequence analysis of upl1 indicated it was 1084 bp long, and encoded a premature protein of 203 amino acids. The N-terminal sequence of the mature UPL1 polypeptide started at amino acid 54 of the deduced sequence from the cDNA, indicating 53 amino acids lost due to posttranslational modification. The primary structure of the Ulva pertusa lectin did not show amino acid sequence similarity with known plant and animal lectins. Hence, this protein may be the paradigm of a novel lectin family.

[Sequence analysis of RNA3 of Rice stripe virus isolates found in China: evidence for reassortment in Tenuivirus].

The RNA3 segments of four isolates of Rice stripe virus (RSV), isolated from endemic sites at Panjin (PJ), Liaoning Province, Kunming (KM) and Yiliang (YL), Yunnan Province, as well as from outbreak sites at Hongze (HZ), Jiangsu Province, were determined. RNA3 of these four isolates were 2480 bp, 2509 bp, 2489 bp and 2497 bp in length, respectively. Compared with RNA3 of T and M isolates from Japan and Y isolate from Yunnan Province of China, that had been previously reported, these seven isolates could be divided into two groups. KM and YL isolates formed group one, and PJ, HZ, Y, T and M isolates belonged to another group. The two groups shared 97%-98% and 93%-94% sequence homology in viral RNA3 (vRNA3) and viral complementary RNA3 (vcRNA3) at the nucleotide levels, respectively, and there was no significant difference between the two groups at the amino acid levels. In the first group, Y isolate was significantly different from HZ,PJ and two Japan isolates in their RNA4 segment. These results show that there were two subgroups in RSV natural population related with geographical location, and reassorment may be the main factor leading to different segments of Y isolate belonging to different subgroups. The results may provide another evidence for reassortment variation in Tenuivirus.

Purification and Characterization of AAVP, a Protein Inhibitor of TMV Infection, from the Edible Fungus, Agrocybe aegerita.

A protein with activity of inhibiting TMV infection on Nicotiana glutinosa, named AAVP, was isolated and purified from the fruiting bodies of edible fungus Agrocybe aegerita by precipitation of 40%--80% saturation of (NH(4))(2)SO(4) followed by DEAE-Fast Flow and S-200 column chromatography. The protein was proved to be homogeneous by SDS-PAGE and IEF. Analysis of the composition of amino acids showed that this protein contained no cysteine, poor in methionine and phenylalanine and rich in acidic and hydroxyl amino acids. The inhibition of TMV was 84.32% when the concentration of AAVP was 200 mg/L. N-terminus of this protein was blocked by pyroglutamyl, and the N-terminal sequence was QGVNIYNIVAGA. On potato-sucrose-agar medium, the purified protein did not inhibit the growth of three plant pathogen fungi, Trichoderma viride, Colletotrichum musae and Fusarium oxysporum.